WorldWideScience

Sample records for surface expression northern

  1. Seasonal prevailing surface winds in Northern Serbia

    Science.gov (United States)

    Tošić, Ivana; Gavrilov, Milivoj B.; Marković, Slobodan B.; Ruman, Albert; Putniković, Suzana

    2018-02-01

    Seasonal prevailing surface winds are analyzed in the territory of Northern Serbia, using observational data from 12 meteorological stations over several decades. In accordance with the general definition of prevailing wind, two special definitions of this term are used. The seasonal wind roses in 16 directions at each station are analyzed. This study shows that the prevailing winds in Northern Serbia have northwestern and southeastern directions. Circulation weather types over Serbia are presented in order to determine the connections between the synoptic circulations and prevailing surface winds. Three controlling pressure centers, i.e., the Mediterranean cyclone, Siberian high, and the Azores anticyclone, appear as the most important large-scale factors that influence the creation of the prevailing winds over Northern Serbia. Beside the synoptic cause of the prevailing winds, it is noted that the orography of the eastern Balkans has a major influence on the winds from the second quadrant. It was found that the frequencies of circulation weather types are in agreement with those of the prevailing winds over Northern Serbia.

  2. Genesis and preservation of a uranium-rich paleozoic epithermal system with a surface expression (Northern Flinders Ranges, South Australia): radiogenic heat driving regional hydrothermal circulation over geological timescales.

    Science.gov (United States)

    Brugger, Joël; Wülser, Pierre-Alain; Foden, John

    2011-01-01

    The surface expressions of hydrothermal systems are prime targets for astrobiological exploration, and fossil systems on Earth provide an analogue to guide this endeavor. The Paleozoic Mt. Gee-Mt. Painter system (MGPS) in the Northern Flinders Ranges of South Australia is exceptionally well preserved and displays both a subsurface quartz sinter (boiling horizon) and remnants of aerial sinter pools that lie in near-original position. The energy source for the MGPS is not related to volcanism but to radiogenic heat produced by U-Th-K-rich host rocks. This radiogenic heat source drove hydrothermal circulation over a long period of time (hundreds of millions of years, from Permian to present), with peaks in hydrothermal activity during periods of uplift and high water supply. This process is reflected by ongoing hot spring activity along a nearby fault. The exceptional preservation of the MGPS resulted from the lack of proximal volcanism, coupled with tectonics driven by an oscillating far-field stress that resulted in episodic basement uplift. Hydrothermal activity caused the remobilization of U and rare earth elements (REE) in host rocks into (sub)economic concentrations. Radiogenic-heat-driven systems are attractive analogues for environments that can sustain life over geological times; the MGPS preserves evidence of episodic fluid flow for the past ∼300 million years. During periods of reduced hydrothermal activity (e.g., limited water supply, quiet tectonics), radiolytic H(2) production has the potential to support an ecosystem indefinitely. Remote exploration for deposits similar to those at the MGPS systems can be achieved by combining hyperspectral and gamma-ray spectroscopy. © Mary Ann Liebert, Inc.

  3. Northern bobwhite breeding season ecology on a reclaimed surface mine

    Science.gov (United States)

    Brooke, Jarred M.; Tanner, Evan P.; Peters, David C.; Tanner, Ashley M.; Harper, Craig A.; Keyser, Patrick D.; Clark, Joseph D.; Morgan, John J.

    2017-01-01

    Surface coal mining and subsequent reclamation of surface mines have converted large forest areas into early successional vegetative communities in the eastern United States. This reclamation can provide a novel opportunity to conserve northern bobwhite (Colinus virginianus). We evaluated the influence of habitat management activities on nest survival, nest-site selection, and brood resource selection on managed and unmanaged units of a reclaimed surface mine, Peabody Wildlife Management Area (Peabody), in west-central Kentucky, USA, from 2010 to 2013. We compared resource selection, using discrete-choice analysis, and nest survival, using the nest survival model in Program MARK, between managed and unmanaged units of Peabody at 2 spatial scales: the composition and configuration of vegetation types (i.e., macrohabitat) and vegetation characteristics at nest sites and brood locations (i.e., microhabitat). On managed sites, we also investigated resource selection relative to a number of different treatments (e.g., herbicide, disking, prescribed fire). We found no evidence that nest-site selection was influenced by macrohabitat variables, but bobwhite selected nest sites in areas with greater litter depth than was available at random sites. On managed units, bobwhite were more likely to nest where herbicide was applied to reduce sericea lespedeza (Lespedeza cuneata) compared with areas untreated with herbicide. Daily nest survival was not influenced by habitat characteristics or by habitat management but was influenced by nest age and the interaction of nest initiation date and nest age. Daily nest survival was greater for older nests occurring early in the breeding season (0.99, SE < 0.01) but was lower for older nests occurring later in the season (0.08, SE = 0.13). Brood resource selection was not influenced by macrohabitat or microhabitat variables we measured, but broods on managed units selected areas treated with herbicide to control sericea lespedeza

  4. The Surface Circulation of the Northern Arabian Sea

    National Research Council Canada - National Science Library

    Kindle, John

    2001-01-01

    Prior to the Arabian Sea Expedition of 1994-1995, researchers' understanding of the northern Arabian Sea's response to the seasonally reversing monsoon cycle was based on climatological ship drift reports...

  5. Stereotypes of emotional expressiveness of northerners and southerners: a cross-cultural test of Montesquieu's hypotheses.

    Science.gov (United States)

    Pennebaker, J W; Rimé, B; Blankenship, V E

    1996-02-01

    Montesquieu argued that residents of warmer climates are more emotionally expressive than those living in cooler ones. More than 2,900 college students from 26 countries completed a brief questionnaire assessing the degree to which they considered Northerners and Southerners within their own countries to be emotionally expressive. In addition, individuals rated themselves on their own degree of expressiveness. In partial confirmation of Montesquieu's hypothesis, it was found that large within-country North-South stereotypes exist. Especially in Old World countries, Northerners are viewed as less emotionally expressive than Southerners. Regression and other analyses revealed that self-ratings of expressiveness were, in fact, related to being from the South and to warmer mean temperatures. Several possible explanations for these effects are discussed.

  6. Relict non-glacial surfaces and autochthonous blockfields in the northern Swedish mountains

    OpenAIRE

    Goodfellow, Bradley W.

    2008-01-01

    Relict non-glacial surfaces occur in many formerly glaciated landscapes, where they represent areas that have escaped significant glacial modification. Frequently distinguished by blockfield mantles, relict non-glacial surfaces are important archives of long-term weathering and landscape evolution processes. The aim of this thesis is to examine the distribution, weathering, ages, and formation of relict non-glacial surfaces in the northern Swedish mountains. Mapping of surfaces from aerial ph...

  7. The European Continent : Surface Expression of Upper Mantle Dynamics

    Science.gov (United States)

    Tondi, M. R.; Schivardi, R.; Molinari, I.; Morelli, A.

    2012-12-01

    images of the European upper mantle isotropic shear-wave speeds and mass densities, recently recovered by combined inversion of surface-wave information and GRACE satellite gravity data (Tondi et al., 2012) are used to select the regions where the residual topography and the residual mantle gravity anomalies are strongly correlated (correlation coefficient is equal to 1). We assume surface uplift processes with negative density anomalies and downward pull with positive anomalies. Our work shows a strong correlation among the areas where, on the basis of our assumptions, the mantle dynamics have surface expression and the areas of low values of radial anisotropy: (1) the southern margins of the East European Craton, (2) the North-Eastern edges of the Arabian Plateau, (3) the northern edge of the CEVP (Central European Volcanic Province), (4) the North-Eastern part of the Atlantic Ocean, between Greenland and Iceland.

  8. The Transit-Time Distribution from the Northern Hemisphere Midlatitude Surface

    Science.gov (United States)

    Orbe, Clara; Waugh, Darryn W.; Newman, Paul A.; Strahan, Susan; Steenrod, Stephen

    2015-01-01

    The distribution of transit times from the Northern Hemisphere (NH) midlatitude surface is a fundamental property of tropospheric transport. Here we present an analysis of the transit time distribution (TTD) since air last contacted the northern midlatitude surface layer, as simulated by the NASA Global Modeling Initiative Chemistry Transport Model. We find that throughout the troposphere the TTD is characterized by long flat tails that reflect the recirculation of old air from the Southern Hemisphere and results in mean ages that are significantly larger than the modal age. Key aspects of the TTD -- its mode, mean and spectral width -- are interpreted in terms of tropospheric dynamics, including seasonal shifts in the location and strength of tropical convection and variations in quasi-isentropic transport out of the northern midlatitude surface layer. Our results indicate that current diagnostics of tropospheric transport are insufficient for comparing model transport and that the full distribution of transit times is a more appropriate constraint.

  9. Changes in the Surface Area of Glaciers in Northern Eurasia

    Science.gov (United States)

    Khromova, T.; Nosenko, G.

    2012-12-01

    Glaciers are widely recognized as key indicators of climate change. Recent evidence suggests an acceleration of glacier mass loss in several key mountain regions. Glacier recession implies the landscape changes in the glacial zone, origin of new lakes and activation of natural disaster processes, catastrophic mudflows, ice avalanches, outburst floods, and etc. The presence of glaciers in itself threats to human life, economic activity and growing infrastructure. Economical and recreational human activity in mountain regions requires relevant information on snow and ice objects. Absence or inadequacy of such information results in financial and human losses. A more comprehensive evaluation of glacier changes is imperative to assess ice contributions to global sea level rise and the future of water resources from glacial basins. One of the urgent steps is a full inventory of all ice bodies, their volume and changes The first estimation of glaciers state and glaciers distribution in the big part of Northern Eurasia has been done in the USSR Glacier Inventory published in 1966 -1980 as a part of IHD activity. The Inventory is based on topographic maps and air photos and reflects the status of the glaciers in 1957-1970y. There is information about 23796 glaciers with area of 78222.3 km2 in the Inventory. It covers 23 glacier systems on Northern Eurasia. In the 80th the USSR Glacier Inventory has been transformed in the digital form as a part of the World Glacier Inventory. Recent satellite data provide a unique opportunity to look again at these glaciers and to evaluate changes in glacier extent for the second part of XX century. In the paper we report about 15 000 glaciers outlines for Caucasus, Pamir, Tien-Shan, Altai, Syntar-Khayata, Cherskogo Range, Kamchatka and Russian Arctic which have been derived from ASTER and Landsat imagery and could be used for glacier changes evaluation. The results show that glaciers are retreating in all these regions. There is, however

  10. Surface Temperatures on Titan During Northern Winter and Spring

    Science.gov (United States)

    Jennings, D. E.; Cottini, V.; Nixon, C. A.; Achterberg, R. K.; Flasar, F. M.; Kunde ,V. G.; Romani, P. N.; Samuelson, R. E.; Mamoutkine, A.; Gorius, N. J. P.; hide

    2016-01-01

    Meridional brightness temperatures were measured on the surface of Titan during the 2004-2014 portion of the Cassini mission by the Composite Infrared Spectrometer. Temperatures mapped from pole to pole during five two year periods show a marked seasonal dependence. The surface temperature near the south pole over this time decreased by 2 K from 91.7 plus or minus 0.3 to 89.7 plus or minus 0.5 K while at the north pole the temperature increased by 1 K from 90.7 plus or minus 0.5 to 91.5 plus or minus 0.2 K. The latitude of maximum temperature moved from 19 S to 16 N, tracking the subsolar latitude. As the latitude changed, the maximum temperature remained constant at 93.65 plus or minus 0.15 K. In 2010 our temperatures repeated the north-south symmetry seen by Voyager one Titan year earlier in 1980. Early in the mission, temperatures at all latitudes had agreed with GCM predictions, but by 2014 temperatures in the north were lower than modeled by 1 K. The temperature rise in the north may be delayed by cooling of sea surfaces and moist ground brought on by seasonal methane precipitation and evaporation.

  11. SURFACE TEMPERATURES ON TITAN DURING NORTHERN WINTER AND SPRING

    Energy Technology Data Exchange (ETDEWEB)

    Jennings, D. E.; Cottini, V.; Nixon, C. A.; Achterberg, R. K.; Flasar, F. M.; Kunde, V. G.; Romani, P. N.; Samuelson, R. E. [Goddard Space Flight Center, Greenbelt, MD 20771 (United States); Mamoutkine, A. [ADNET Systems, Inc., Bethesda, MD 20817 (United States); Gorius, N. J. P. [The Catholic University of America, Washington, DC 20064 (United States); Coustenis, A. [Laboratoire d’Etudes Spatiales et d’Instrumentation en Astrophysique (LESIA), Observatoire de Paris, CNRS, UPMC Univ. Paris 06, Univ. Paris-Diderot, 5, place Jules Janssen, F-92195 Meudon Cedex (France); Tokano, T., E-mail: donald.e.jennings@nasa.gov [Universität zu Köln, Albertus-Magnus-Platz, D-50923 Köln (Germany)

    2016-01-01

    Meridional brightness temperatures were measured on the surface of Titan during the 2004–2014 portion of the Cassini mission by the Composite Infrared Spectrometer. Temperatures mapped from pole to pole during five two-year periods show a marked seasonal dependence. The surface temperature near the south pole over this time decreased by 2 K from 91.7 ± 0.3 to 89.7 ± 0.5 K while at the north pole the temperature increased by 1 K from 90.7 ± 0.5 to 91.5 ± 0.2 K. The latitude of maximum temperature moved from 19 S to 16 N, tracking the sub-solar latitude. As the latitude changed, the maximum temperature remained constant at 93.65 ± 0.15 K. In 2010 our temperatures repeated the north–south symmetry seen by Voyager one Titan year earlier in 1980. Early in the mission, temperatures at all latitudes had agreed with GCM predictions, but by 2014 temperatures in the north were lower than modeled by 1 K. The temperature rise in the north may be delayed by cooling of sea surfaces and moist ground brought on by seasonal methane precipitation and evaporation.

  12. Chemistry of the sea-surface microlayer. 3. Studies on the nutrient chemistry of the northern Arabian Sea

    Digital Repository Service at National Institute of Oceanography (India)

    Singbal, S.Y.S.; Narvekar, P.V.; Nagarajan, R.

    Nutrients showed enrichment in the surface microlayer compared to those in sub-surface water and there was a decreasing trend in the enrichment factor from nearshore to offshore in Northern Arabian Sea. The nutrient concentrations were correlated...

  13. Geomorphic Surface Maps of Northern Frenchman Flat, Nevada Test Site, Southern Nevada

    International Nuclear Information System (INIS)

    Bechtel Nevada

    2005-01-01

    Large-scale (1:6000) surficial geology maps of northern Frenchman Flat were developed in 1995 as part of comprehensive site characterization required to operate a low-level radioactive waste disposal facility in that area. Seven surficial geology maps provide fundamental data on natural processes and are the platform needed to reconstruct the Quaternary history of northern Frenchman Flat. Reconstruction of the Quaternary history provides an understanding of the natural processes that act to develop the landscape, and the time-frames involved in landscape development. The mapping was conducted using color and color-infrared aerial photographs and field verification of map unit composition and boundaries. Criteria for defining the map unit composition of geomorphic surface units are based on relative geomorphic position, landform morphology, and degree of preservation of surface morphology. Seven geomorphic surfaces (Units 1 through 7) are recognized, spanning from the early Quaternary to present time

  14. EAARL Coastal Topography - Northern Gulf of Mexico, 2007: First Surface

    Science.gov (United States)

    Smith, Kathryn E.L.; Nayegandhi, Amar; Wright, C. Wayne; Bonisteel, Jamie M.; Brock, John C.

    2009-01-01

    These remotely sensed, geographically referenced elevation measurements of Lidar-derived first surface (FS) elevation data were produced as a collaborative effort between the U.S. Geological Survey (USGS), Florida Integrated Science Center (FISC), St. Petersburg, FL; the National Park Service (NPS), Gulf Coast Network, Lafayette, LA; and the National Aeronautics and Space Administration (NASA), Wallops Flight Facility, VA. The project provides highly detailed and accurate datasets of select barrier islands and peninsular regions of Louisiana, Mississippi, Alabama, and Florida, acquired June 27-30, 2007. The datasets are made available for use as a management tool to research scientists and natural resource managers. An innovative airborne Lidar instrument originally developed at the NASA Wallops Flight Facility, and known as the Experimental Advanced Airborne Research Lidar (EAARL), was used during data acquisition. The EAARL system is a raster-scanning, waveform-resolving, green-wavelength (532-nanometer) Lidar designed to map near-shore bathymetry, topography, and vegetation structure simultaneously. The EAARL sensor suite includes the raster-scanning, water-penetrating full-waveform adaptive Lidar, a down-looking red-green-blue (RGB) digital camera, a high-resolution multi-spectral color infrared (CIR) camera, two precision dual-frequency kinematic carrier-phase GPS receivers, and an integrated miniature digital inertial measurement unit which provide for submeter georeferencing of each laser sample. The nominal EAARL platform is a twin-engine Cessna 310 aircraft, but the instrument may be deployed on a range of light aircraft. A single pilot, a Lidar operator, and a data analyst constitute the crew for most survey operations. This sensor has the potential to make significant contributions in measuring sub-aerial and submarine coastal topography within cross-environmental surveys. Elevation measurements were collected over the survey area using the EAARL system

  15. Northern South China Sea Surface Circulation and its Variability Derived by Combining Satellite Altimetry and Surface Drifter Data

    Directory of Open Access Journals (Sweden)

    N. Peter Benny

    2015-01-01

    Full Text Available The present study analyses the mean and seasonal mesoscale surface circulation of the Northern South China Sea (NSCS and determines the influence of El Niño/SouthernNiño/Southern Oscillation (ENSO. High resolution Eulerian velocity field is derived by combining the available satellite tracked surface drifter data with satellite altimetry during 1993 - 2012. The wind driven current is computed employing the weekly ocean surface mean wind fields derived from the scatterometers on board ERS 1/2, QuikSCAT and ASCAT. The derived mean velocity field exhibits strong boundary currents and broad zonal flow across NSCS. The anomalous field is quite strong in the southern part and the Seasonal circulation clearly depicts the monsoonal forcing. Eddy Kinetic Energy (EKE distribution and its spatial and temporal structures are determined employing Empirical Orthogonal Function (EOF analysis. The ENSO influence on NSCS surface circulation has been analyzed using monthly absolute geostrophic velocity fields during 1996 - 1999.

  16. Active tectonics revealed by isobase surfaces analysis from South Rifian Ridges, Northern Morocco

    OpenAIRE

    Amine, Afaf; El Ouardi, Hmidou

    2017-01-01

    The analysis of isobase maps, as one of surface dynamic parameters , helps understanding the tectonic geomorphology of South Rifian Ridges, Northern Morocco . The automatic generation of morphostructural map of 2nd and 3rd order channels is based on 30-grid Digital Elevation Modelin to Geographic Information System environment presents good results, supporting previous studies. Comparing to the western ridges, the eastern ones, specially Moulay Idriss massif, evidence neotectonic activity tri...

  17. Sea-surface salinity variations in the northern Caribbean Sea across the Mid-Pleistocene Transition

    OpenAIRE

    Sepulcre, S.; Vidal, L.; Tachikawa, K.; Rostek, F.; Bard, E.

    2011-01-01

    By reconstructing past hydrologic variations in the Northern Caribbean Sea and their influence on the stability of the Atlantic Meridional Overturning Circulation (AMOC) during the last 940 ka, we seek to document climate changes in this tropical area in response to the Mid-Pleistocene Transition (MPT). Using core MD03-2628, we estimated past changes in sea surface salinity (SSS) using Δδ18O, the difference between the modern, and the past &delta...

  18. Hard-bottom succession of subtidal epibenthic communities colonizing hidden and exposed surfaces off northern Chile

    Directory of Open Access Journals (Sweden)

    Aldo S. Pacheco

    2010-03-01

    Full Text Available The biodiversity of hard-bottom substrata comprises species growing on exposed rock and in hidden microhabitats, such as cracks and crevices. This study examines the succession of epibenthic organisms colonizing an artificial substratum with one surface exposed and one surface hidden on a vertical wall off northern Chile. On each sampling date species coverage of three replicate panels on both surfaces was assessed. The hidden surface was dominated in terms of coverage by the bryozoans Membranipora isabelleana and Lagenicella variabilis, while algae were absent. In contrast, the exposed surface was dominated by encrusting red corallines and the red alga Rhodymenia corallina. At the end of the experimental period both surfaces were dominated by colonial suspension feeders, but showed a different community structure and successional pattern. On the exposed surface, competitive exclusion was identified as an important aspect of succession, whereas on the hidden surface this pattern was not observed. These findings have implications for overall biodiversity, because pioneer species that are not able to survive long periods on exposed surfaces become restricted to hidden surfaces, from where they spread laterally. Thus, hidden microhabitats provide refuges for certain species, and may play an important role in the overall succession on rock faces. We conclude that examination of hidden microhabitats is necessary in order to fully understand succession in hard-bottom habitats.

  19. Tracking the complete revolution of surface westerlies over Northern Hemisphere using radionuclides emitted from Fukushima

    International Nuclear Information System (INIS)

    Hernández-Ceballos, M.A.; Hong, G.H.; Lozano, R.L.; Kim, Y.I.; Lee, H.M.; Kim, S.H.; Yeh, S.-W.; Bolívar, J.P.; Baskaran, M.

    2012-01-01

    Massive amounts of anthropogenic radionuclides were released from the nuclear reactors located in Fukushima (northeastern Japan) between 12 and 16 March 2011 following the earthquake and tsunami. Ground level air radioactivity was monitored around the globe immediately after the Fukushima accident. This global effort provided a unique opportunity to trace the surface air mass movement at different sites in the Northern Hemisphere. Based on surface air radioactivity measurements around the globe and the air mass backward trajectory analysis of the Fukushima radioactive plume at various places in the Northern Hemisphere by employing the Hybrid Single-Particle Lagrangian Integrated Trajectory model, we show for the first time, that the uninterrupted complete revolution of the mid-latitude Surface Westerlies took place in less than 21 days, with an average zonal velocity of > 60 km/h. The position and circulation time scale of Surface Westerlies are of wide interest to a large number of global researchers including meteorologists, atmospheric researchers and global climate modellers. -- Highlights: ► Evidence of the South Korea contamination with released radiocesium from Fukushima. ► Field samples and air mass analysis were utilized to elucidate the transport of those radionuclides. ► Characterization of the air mass movements at different sites at the Earth's surface. ► Verification of the uninterrupted complete revolution of the artificial radionuclides released in Fukushima. ► Quantification of the velocity of the artificial radionuclides released in Fukushima.

  20. Response of the global surface ozone distribution to Northern Hemisphere sea surface temperature changes: implications for long-range transport

    Science.gov (United States)

    Yi, Kan; Liu, Junfeng; Ban-Weiss, George; Zhang, Jiachen; Tao, Wei; Cheng, Yanli; Tao, Shu

    2017-07-01

    The response of surface ozone (O3) concentrations to basin-scale warming and cooling of Northern Hemisphere oceans is investigated using the Community Earth System Model (CESM). Idealized, spatially uniform sea surface temperature (SST) anomalies of ±1 °C are individually superimposed onto the North Pacific, North Atlantic, and North Indian oceans. Our simulations suggest large seasonal and regional variability in surface O3 in response to SST anomalies, especially in the boreal summer. The responses of surface O3 associated with basin-scale SST warming and cooling have similar magnitude but are opposite in sign. Increasing the SST by 1 °C in one of the oceans generally decreases the surface O3 concentrations from 1 to 5 ppbv. With fixed emissions, SST increases in a specific ocean basin in the Northern Hemisphere tend to increase the summertime surface O3 concentrations over upwind regions, accompanied by a widespread reduction over downwind continents. We implement the integrated process rate (IPR) analysis in CESM and find that meteorological O3 transport in response to SST changes is the key process causing surface O3 perturbations in most cases. During the boreal summer, basin-scale SST warming facilitates the vertical transport of O3 to the surface over upwind regions while significantly reducing the vertical transport over downwind continents. This process, as confirmed by tagged CO-like tracers, indicates a considerable suppression of intercontinental O3 transport due to increased tropospheric stability at lower midlatitudes induced by SST changes. Conversely, the responses of chemical O3 production to regional SST warming can exert positive effects on surface O3 levels over highly polluted continents, except South Asia, where intensified cloud loading in response to North Indian SST warming depresses both the surface air temperature and solar radiation, and thus photochemical O3 production. Our findings indicate a robust linkage between basin-scale SST

  1. Response of the global surface ozone distribution to Northern Hemisphere sea surface temperature changes: implications for long-range transport

    Directory of Open Access Journals (Sweden)

    K. Yi

    2017-07-01

    Full Text Available The response of surface ozone (O3 concentrations to basin-scale warming and cooling of Northern Hemisphere oceans is investigated using the Community Earth System Model (CESM. Idealized, spatially uniform sea surface temperature (SST anomalies of ±1 °C are individually superimposed onto the North Pacific, North Atlantic, and North Indian oceans. Our simulations suggest large seasonal and regional variability in surface O3 in response to SST anomalies, especially in the boreal summer. The responses of surface O3 associated with basin-scale SST warming and cooling have similar magnitude but are opposite in sign. Increasing the SST by 1 °C in one of the oceans generally decreases the surface O3 concentrations from 1 to 5 ppbv. With fixed emissions, SST increases in a specific ocean basin in the Northern Hemisphere tend to increase the summertime surface O3 concentrations over upwind regions, accompanied by a widespread reduction over downwind continents. We implement the integrated process rate (IPR analysis in CESM and find that meteorological O3 transport in response to SST changes is the key process causing surface O3 perturbations in most cases. During the boreal summer, basin-scale SST warming facilitates the vertical transport of O3 to the surface over upwind regions while significantly reducing the vertical transport over downwind continents. This process, as confirmed by tagged CO-like tracers, indicates a considerable suppression of intercontinental O3 transport due to increased tropospheric stability at lower midlatitudes induced by SST changes. Conversely, the responses of chemical O3 production to regional SST warming can exert positive effects on surface O3 levels over highly polluted continents, except South Asia, where intensified cloud loading in response to North Indian SST warming depresses both the surface air temperature and solar radiation, and thus photochemical O3 production. Our findings indicate a robust linkage

  2. Hard boys, attractive girls: expressions of gender in young people's conversations on smoking in Northern Ireland.

    Science.gov (United States)

    Rugkåsa, Jorun; Stewart-Knox, Barbara; Sittlington, Julie; Abaunza, Pilar Santos; Treacy, Margaret P

    2003-12-01

    Health behaviour and attitudes among young people can be interpreted within the context of personal and social identity. This paper explores ways in which 10- to 11-year-olds in Northern Ireland expressed perceptions of gender ideology while discussing the topic of smoking. The data were gathered by means of in-depth interviews (n=85). Although few had tried smoking, the positive quality of their own gender appeared to be expressed through negative assessment of smokers of the opposite gender, and on different ideas of perceived gender dimensions in adult smoking. In order to deliver successful health promotion interventions, it is crucial to understand and address differences based on gender as it may partly explain differences in smoking experimentation and prevalence.

  3. Less sensitive of urban surface to climate variability than rural in Northern China.

    Science.gov (United States)

    Yao, Rui; Wang, Lunche; Huang, Xin; Chen, Jiangping; Li, Jiarui; Niu, Zigeng

    2018-02-14

    In this study, the relationships between interannual variations of surface urban heat islands (SUHIs) and climate variability were studied in 31 cities of China for the period 2001-2016. For cold and dry Northern China, it was found that the interannual variations of SUHI intensity (SUHII, land surface temperature (LST) in urban minus rural) in urban cores was significantly (purban cores (1.141°C for SDs and 2.535°C for WDs) than in rural areas (1.890°C for SDs and 3.377°C for WDs). The standard deviation was further used to reflect the interannual stabilities of LST, enhanced vegetation index (EVI) and white sky albedo (WSA). Interestingly, the standard deviations of LST across 2001-2016 were generally lower in urban cores (0.994°C for SDs and 1.577°C for WDs) than in rural areas (1.431°C for SDs and 2.077°C for WDs). Similar results were observed for EVI and WSA (winter). The results suggested that the urban surface is less sensitive to climate variability than rural areas in Northern China. Comparatively, most findings were less evident in hot and humid Southern China. Despite the whole world would become warmer or colder in future, the insensitivity of urban surface may mitigate its impacts in cold and dry Northern China. However, it does not mean that urbanization is totally good due to its environmental problem. Copyright © 2018 Elsevier B.V. All rights reserved.

  4. Using Land Surface Phenology to Detect Land Use Change in the Northern Great Plains

    Science.gov (United States)

    Nguyen, L. H.; Henebry, G. M.

    2017-12-01

    The Northern Great Plains of the US have been undergoing many types of land cover / land use change over the past two decades, including expansion of irrigation, conversion of grassland to cropland, biofuels production, urbanization, and fossil fuel mining. Much of the literature on these changes has relied on post-classification change detection based on a limited number of observations per year. Here we demonstrate an approach to characterize land dynamics through land surface phenology (LSP) by synergistic use of image time series at two scales. Our study areas include regions of interest (ROIs) across the Northern Great Plains located within Landsat path overlap zones to boost the number of valid observations (free of clouds or snow) each year. We first compute accumulated growing degree-days (AGDD) from MODIS 8-day composites of land surface temperature (MOD11A2 and MYD11A2). Using Landsat Collection 1 surface reflectance-derived vegetation indices (NDVI, EVI), we then fit at each pixel a downward convex quadratic model linking the vegetation index to each year's progression of AGDD. This quadratic equation exhibits linearity in a mathematical sense; thus, the fitted models can be linearly mixed and unmixed using a set of LSP endmembers (defined by the fitted parameter coefficients of the quadratic model) that represent "pure" land cover types with distinct seasonal patterns found within the region, such as winter wheat, spring wheat, maize, soybean, sunflower, hay/pasture/grassland, developed/built-up, among others. Information about land cover corresponding to each endmember are provided by the NLCD (National Land Cover Dataset) and CDL (Cropland Data Layer). We use linear unmixing to estimate the likely proportion of each LSP endmember within particular areas stratified by latitude. By tracking the proportions over the 2001-2011 period, we can quantify various types of land transitions in the Northern Great Plains.

  5. Quality assessment of spaceborne sea surface salinity observations over the northern North Atlantic

    Science.gov (United States)

    Köhler, Julia; Sena Martins, Meike; Serra, Nuno; Stammer, Detlef

    2015-01-01

    Spaceborne sea surface salinity (SSS) measurements provided by the European Space Agency's (ESA) "Soil Moisture and Ocean Salinity" (SMOS) and the National Aeronautical Space Agency's (NASA) "Aquarius/SAC-D" missions, covering the period from May 2012 to April 2013, are compared against in situ salinity measurements obtained in the northern North Atlantic between 20°N and 80°N. In cold water, SMOS SSS fields show a temperature-dependent negative SSS bias of up to -2 g/kg for temperatures associated sampling errors there.

  6. Increased liver apoptosis and tumor necrosis factor expression in Atlantic bluefin tuna (Thunnus thynnus) reared in the northern Adriatic Sea.

    Science.gov (United States)

    Corriero, Aldo; Zupa, Rosa; Pousis, Chrysovalentinos; Santamaria, Nicoletta; Bello, Giambattista; Jirillo, Emilio; Carrassi, Michele; De Giorgi, Carla; Passantino, Letizia

    2013-06-15

    The Atlantic bluefin tuna Thunnus thynnus (ABFT) is intensely fished in the Mediterranean Sea to supply a prosperous capture-based mariculture industry. Liver apoptotic structures and tumor necrosis factor (TNF) gene expression were determined in: wild ABFT caught in the eastern Atlantic; juvenile ABFT reared in the central Adriatic Sea; juvenile ABFT reared in the northern Adriatic Sea; adult ABFT reared in the western Mediterranean. The highest density of liver apoptotic structures was found in the juveniles from the northern Adriatic. Two partial TNF cDNAs (TNF1 and TNF2) were cloned and sequenced. TNF1 gene expression was higher in juveniles than in adults. The highest expression of TNF2 was found in the juveniles from the northern Adriatic. These findings might be related to the juvenile exposure to environmental pollutants. Copyright © 2013 Elsevier Ltd. All rights reserved.

  7. Natural sunlight shapes crude oil-degradingbacterial communities in northern Gulf of Mexico surface waters

    Directory of Open Access Journals (Sweden)

    Hernando P Bacosa

    2015-12-01

    Full Text Available Following the Deepwater Horizon (DWH spill in 2010, an enormous amount of oil was observed in the deep and surface waters of the northern Gulf of Mexico. Surface waters are characterized by intense sunlight and high temperature during summer. While the oil-degrading bacterial communities in the deep-sea plume have been widely investigated, the effect of natural sunlight on those in oil polluted surface waters remains unexplored to date. In this study, we incubated surface water from the DWH site with amendments of crude oil, Corexit dispersant, or both for 36 d under natural sunlight in the northern Gulf of Mexico. The bacterial community was analyzed over time for total abundance, density of alkane and polycyclic aromatic hydrocarbon degraders, and community composition via pyrosequencing. Our results showed that, for treatments with oil and/or Corexit, sunlight significantly reduced bacterial diversity and evenness and was a key driver of shifts in bacterial community structure. In samples containing oil or dispersant, sunlight greatly reduced abundance of the Cyanobacterium Synechococcus but increased the relative abundances of Alteromonas, Marinobacter, Labrenzia, Sandarakinotalea, Bartonella, and Halomonas. Dark samples with oil were represented by members of Thalassobius, Winogradskyella, Alcanivorax, Formosa, Pseudomonas, Eubacterium, Erythrobacter, Natronocella, and Coxiella. Both oil and Corexit inhibited the Candidatus Pelagibacter with or without sunlight exposure. For the first time, we demonstrated the effects of light in structuring microbial communities in water with oil and/or Corexit. Overall, our findings improve understanding of oil pollution in surface water, and provide unequivocal evidence that sunlight is a key factor in determining bacterial community composition and dynamics in oil polluted marine waters.

  8. Crustal structure of northern Egypt from joint inversion of receiver functions and surface wave dispersion velocities

    Science.gov (United States)

    Badawy, Ahmed; Hegazi, Mona; Gaber, Hanan; Korrat, Ibrahim

    2018-01-01

    In this study, we used a combined inversion of body wave receiver functions and surface wave dispersion measurements to provide constraints on the crustal structure of northern Egypt. The two techniques are complementary to each other: receiver functions (RFs) are sensitive to shear-wave velocity contrasts, while surface wave dispersion (SWD) measurements are sensitive to finite variations of shear-wave velocity with depth. A database of 122 teleseismic events digitally recorded by the Egyptian National Seismological Network (ENSN) stations has been used as well. To enhance the resulting RFs at each ENSN station, the H-k stacking method was applied. A joint inversion process between the resulting receiver functions and the surface wave dispersion curves was applied as well. We have produced three averaged velocity structure models for distinct geographic and tectonic provinces namely Sinai, eastern desert, and western desert from east to the west respectively. These models will deeply help in estimation the epicenter distance of earthquake, focal mechanism solutions, and earthquake hazard analysis in northern Egypt. An obvious image of the subsurface structure has been determined which shows that generally the crustal structure of northern Egypt consists of three layers covered with a sequence of sediments that differs in thickness from across the region except in the Sharm area where the sedimentary cover is absent. The obtained results indicate that crustal thickness differs from east to west and reaches its maximum value of about 36 km at Siwa station (SWA) in the western desert and its minimum value of about 28 km at Sharm station (SHR) of the southern tip of the Sinai Peninsula. The Vp/Vs ratio varies between 1.71 and 2.07 in northern Egypt. Generally, the high values (1.93) of (Vp/Vs) at SWA station may reflect the well-known rich aquifer with fully saturated sediments of the Swia Oasis in the Western Desert. Moreover, the highest value (2.07) of (Vp/Vs) at

  9. The surface energy balance of a polygonal tundra site in northern Siberia – Part 2: Winter

    Directory of Open Access Journals (Sweden)

    J. Boike

    2011-06-01

    Full Text Available In this study, we present the winter time surface energy balance at a polygonal tundra site in northern Siberia based on independent measurements of the net radiation, the sensible heat flux and the ground heat flux from two winter seasons. The latent heat flux is inferred from measurements of the atmospheric turbulence characteristics and a model approach. The long-wave radiation is found to be the dominant factor in the surface energy balance. The radiative losses are balanced to about 60 % by the ground heat flux and almost 40 % by the sensible heat fluxes, whereas the contribution of the latent heat flux is small. The main controlling factors of the surface energy budget are the snow cover, the cloudiness and the soil temperature gradient. Large spatial differences in the surface energy balance are observed between tundra soils and a small pond. The ground heat flux released at a freezing pond is by a factor of two higher compared to the freezing soil, whereas large differences in net radiation between the pond and soil are only observed at the end of the winter period. Differences in the surface energy balance between the two winter seasons are found to be related to differences in snow depth and cloud cover which strongly affect the temperature evolution and the freeze-up at the investigated pond.

  10. Molecular expression of aquaporin mRNAs in the northern house mosquito, Culex pipiens.

    Science.gov (United States)

    Yang, Liu; Piermarini, Peter M

    2017-01-01

    Aquaporins (AQPs) are a family of membrane-bound proteins, originally described as water-channels, that broadly exist in prokaryotic and eukaryotic cells. However, some AQPs can transport small molecules (e.g. urea, glycerol) along with or preferentially to water. Previous work in the mosquitoes Aedes aegypti and Anopheles gambiae have characterized the molecular expression of one or more AQP genes and shown that they are involved in water homeostasis after blood feeding, tolerance to dehydration and heat stresses, and development of Plasmodium falciparum in the mosquito. However, the molecular expression of AQPs in the northern house mosquito Culex pipiens, the primary vector of West Nile virus, is presently unknown. Here we characterize the relative mRNA expression of six AQP genes in C. pipiens. We used quantitative RT-PCR (qPCR) to compare AQP mRNA levels 1) among different life stages and sexes, 2) between blood fed and non-blood fed adult females, 3) between adult females injected with phosphate-buffered saline (PBS) and non-injected females, and 4) between adult females exposed to a low vs. high relative humidity. We found that: 1) four AQP mRNAs (Drip, Bib, Eglp1, Aqp12L) were differentially expressed between larval and adult life stages, but not between sexes; 2) after a blood meal, the expression of two AQP mRNAs (Drip and Aqp12L) were down-regulated and one AQP mRNA (Eglp2) was up-regulated; 3) the expression of two AQP mRNAs (Drip and Prip) were down-regulated and one AQP mRNA (Eglp1) was up regulated after injecting the hemolymph of mosquitoes with PBS; and 4) the expression of Prip and Eglp2 mRNAs were both down-regulated in response to a low relative humidity. Our study is the first to characterize the molecular expression patterns of AQPs in C. pipiens, thus providing a foundation for future research elucidating the physiological and functional roles of AQPs in this important disease vector. Copyright © 2016 Elsevier Ltd. All rights reserved.

  11. Downregulation of transferrin receptor surface expression by intracellular antibody

    International Nuclear Information System (INIS)

    Peng Jilin; Wu Sha; Zhao Xiaoping; Wang Min; Li Wenhan; Shen Xin; Liu Jing; Lei Ping; Zhu Huifen; Shen Guanxin

    2007-01-01

    To deplete cellular iron uptake, and consequently inhibit the proliferation of tumor cells, we attempt to block surface expression of transferrin receptor (TfR) by intracellular antibody technology. We constructed two expression plasmids (scFv-HAK and scFv-HA) coding for intracellular single-chain antibody against TfR with or without endoplasmic reticulum (ER) retention signal, respectively. Then they were transfected tumor cells MCF-7 by liposome. Applying RT-PCR, Western blotting, immunofluorescence microscopy and immunoelectron microscope experiments, we insure that scFv-HAK intrabody was successfully expressed and retained in ER contrasted to the secreted expression of scFv-HA. Flow cytometric analysis confirmed that the TfR surface expression was markedly decreased approximately 83.4 ± 2.5% in scFv-HAK transfected cells, while there was not significantly decrease in scFv-HA transfected cells. Further cell growth and apoptosis characteristics were evaluated by cell cycle analysis, nuclei staining and MTT assay. Results indicated that expression of scFv-HAK can dramatically induce cell cycle G1 phase arrest and apoptosis of tumor cells, and consequently significantly suppress proliferation of tumor cells compared with other control groups. For First time this study demonstrates the potential usage of anti-TfR scFv-intrabody as a growth inhibitor of TfR overexpressing tumors

  12. Effects of two decades of rising sea surface temperatures on sublittoral macrobenthos communities in Northern Ireland, UK.

    Science.gov (United States)

    Goodwin, Claire E; Strain, Elisabeth M A; Edwards, Hugh; Bennett, Stephanie C; Breen, Joe P; Picton, Bernard E

    2013-04-01

    We examined whether two decades of rising sea surface temperatures have resulted in significant changes in the benthic community and frequency of occurrence of Northern and Southern species in three areas of Northern Ireland, using visual census data collected by SCUBA surveys undertaken during two periods: pre-1986 and post-2006. We found little evidence to suggest that rising sea surface temperatures have contributed to the changes in benthic assemblage structure between the pre-1986 and post-2006 surveys. However, there were slight but not significant declines in extreme Northern species at Rathlin Island, and increases in the mean number and frequency of occurrence of extreme Southern species in all three areas. There were also substantial declines in the spatial presence of 7 extreme Northern species and notable increases in distribution of 19 extreme Southern species. In contrast, there were no clear trends in the intermediate to Northern and intermediate to Southern species. These results suggest that rising sea surface temperatures have had significant impacts on the occurrence of rarer marine invertebrate species at the edges of their biogeographic range however the trends differed between areas in Northern Ireland. Copyright © 2012 Elsevier Ltd. All rights reserved.

  13. Evaluation of differential gene expression during behavioral development in the honeybee using microarrays and northern blots

    Science.gov (United States)

    Kucharski, Robert; Maleszka, Ryszard

    2002-01-01

    Background The honeybee (Apis mellifera) has been used with great success in a variety of behavioral studies. The lack of genomic tools in this species has, however, hampered efforts to provide genome-based explanations for behavioral data. We have combined the power of DNA arrays and the availability of distinct behavioral stages in honeybees to explore the dynamics of gene expression during adult development in this insect. In addition, we used caffeine treatment, a procedure that accelerates learning abilities in honeybees, to examine changes in gene expression underlying drug-induced behavioral modifications. Results Spotted microarrays containing several thousand cDNAs were interrogated with RNAs extracted from newly emerged worker bees, experienced foragers and caffeine-treated bees. Thirty-six differentially expressed cDNAs were verified by northern blot hybridization and characterized in silico by sequencing and database searches. Experienced foragers overexpressed royal jelly proteins, a putative imaginal disc growth factor, a transcriptional regulator (Stck) and several enzymes, including α-glucosidases, aminopeptidases and glucose dehydrogenase. Naive workers showed increased expression of members of the SPARC and lectin families, heat-shock cognate proteins and several proteins related to RNA translation and mitochondrial function. A number of novel genes overexpressed in both naive and experienced bees, and genes induced by caffeine, have also been identified. Conclusions We have shown the usefulness of this transcriptome-based approach for gene discovery, in particular in the context of the efficacy of drug treatment, in a model organism in which routine genetic techniques cannot be applied easily. PMID:11864369

  14. Seasonal variation in surface fuel moisture between unthinned and thinned mixed conifer forest, northern California, USA

    Science.gov (United States)

    Becky L. Estes; Eric E. Knapp; Carl N. Skinner; Fabian C. C. Uzoh

    2012-01-01

    Reducing stand density is often used as a tool for mitigating the risk of high-intensity crown fires. However, concern has been expressed that opening stands might lead to greater drying of surface fuels, contributing to increased fire risk. The objective of this study was to determine whether woody fuel moisture differed between unthinned and thinned mixed-conifer...

  15. Surface Water Quality Survey of Northern Indian River Lagoon from Sebastian Inlet to Mosquito Lagoon

    Science.gov (United States)

    Weaver, R. J.; Webb, B. M.

    2012-12-01

    Following news of an emerging brown tide algal bloom in the northern Indian River Lagoon (IRL), researchers sought to gain insight into the surface water quality in the IRL, as well as the extent of the algae coverage. A Portable SeaKeeper from YSI, mounted to a personal watercraft-based coastal profiling system, autonomously collected and analyzed the surface water. The system operates by recording sample data every 12 seconds while continuously underway at speeds up to and greater than 50 km/hr. The researchers covered a transect that started at Sebastian Inlet and followed a zig-zag path extending up through the Haulover Canal and into the Mosquito Lagoon. The survey path covered 166.7 km, and collected 2248 samples. Along the way stops were made at water quality stations used by the Saint John's River Water Management District, so that the data collected can be incorporated into ongoing monitoring efforts. The system analyzed the surface water for dissolved oxygen, pH, chlorophyll-a, salinity, temperature, turbidity, refined fuels, and CDOM. In the two days following the lagoon survey, the inlets at Port Canaveral and Sebastian were also surveyed for tidal currents and hydrography. The IRL transect survey data recorded evidence of the southern extent of the algae bloom in both chlorophyll-a and pH levels. Visual evidence of the bloom was striking as the water in the northern IRL turned a milk chocolaty brown color. Chlorophyll-a levels in the two inlets suggested bloom activity at these locations; however this bloom was different. This oceanic bloom was a result of a persistent upwelling event along the East Florida shelf, and the color was a paler green-yellow. The near-synoptic nature of the comprehensive lagoon survey, conducted in just over 7 hours, allows researchers to obtain a better understanding of water quality in coastal lagoons. Elevated levels of salinity, temperature, and refined fuels in the northern IRL indicate a low exchange rate and absence

  16. Paired Expression Analysis of Tumor Cell Surface Antigens

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    Rimas J. Orentas

    2017-08-01

    Full Text Available Adoptive immunotherapy with antibody-based therapy or with T cells transduced to express chimeric antigen receptors (CARs is useful to the extent that the cell surface membrane protein being targeted is not expressed on normal tissues. The most successful CAR-based (anti-CD19 or antibody-based therapy (anti-CD20 in hematologic malignancies has the side effect of eliminating the normal B cell compartment. Targeting solid tumors may not provide a similar expendable marker. Beyond antibody to Her2/NEU and EGFR, very few antibody-based and no CAR-based therapies have seen broad clinical application for solid tumors. To expand the way in which the surfaceome of solid tumors can be analyzed, we created an algorithm that defines the pairwise relative overexpression of surface antigens. This enables the development of specific immunotherapies that require the expression of two discrete antigens on the surface of the tumor target. This dyad analysis was facilitated by employing the Hotelling’s T-squared test (Hotelling–Lawley multivariate analysis of variance for two independent variables in comparison to a third constant entity (i.e., gene expression levels in normal tissues. We also present a unique consensus scoring mechanism for identifying transcripts that encode cell surface proteins. The unique application of our bioinformatics processing pipeline and statistical tools allowed us to compare the expression of two membrane protein targets as a pair, and to propose a new strategy based on implementing immunotherapies that require both antigens to be expressed on the tumor cell surface to trigger therapeutic effector mechanisms. Specifically, we found that, for MYCN amplified neuroblastoma, pairwise expression of ACVR2B or anaplastic lymphoma kinase (ALK with GFRA3, GFRA2, Cadherin 24, or with one another provided the strongest hits. For MYCN, non-amplified stage 4 neuroblastoma, neurotrophic tyrosine kinase 1, or ALK paired with GFRA2, GFRA3, SSK

  17. Regional velocity structure in northern California from inversion of scattered seismic surface waves

    Science.gov (United States)

    Pollitz, Fred F.

    1999-07-01

    Seismic surface waves recorded by the Berkeley Digital Seismic Network have been analyzed in order to constrain three-dimensional lateral heterogeneity of the upper mantle under northern California. A total of 2164 seismograms from 173 teleseismic events were windowed for the fundamental mode Rayleigh wave, followed by estimation of complex amplitude spectra over the period range 16 to 100 s using a multiple-taper method. Since Rayleigh waves at shorter periods, particularly below 35 s, suffer from serious multipathing or "non-plane" wave arrivals, these amplitude spectra have been interpreted as the product of wavefront distortion along the teleseismic propagation path and seismic structure beneath the network. The amplitude spectra are first modeled in terms of non-plane incoming wavefields and structural phase velocity perturbations period by period. After corrections for Moho and surface topography, the phase velocity maps are inverted for three-dimensional shear velocity perturbations δνs down to a depth of 200 km. The δνs maps are in good agreement with the results of body studies over a broad spatial scale. The dominant signals are associated with the thermal effects of the active Gorda and fossil Farallon subducted slab stretching from Mount Shasta through the western Sierran foothills to the southern Great Valley and asthenospheric upwelling beneath the northern Coast Ranges. The southern Sierra Nevada Range is characterized by fast δνs down to ˜50 km and slow velocities between ˜60 and 120 km depth, in agreement with independent inferences of a cold crust and warm upper mantle, which may provide the buoyancy forces necessary to support the elevation of the range.

  18. Responses of energy partitioning and surface resistance to drought in a poplar plantation in northern China

    Science.gov (United States)

    Kang, M.; Zhang, Z.; Noormets, A.; Fang, X.; Zha, T.; Zhou, J.; Sun, G.; McNulty, S.; Chen, J.

    2015-01-01

    Poplar (Populus sp.) plantations have been used broadly for combating desertification, urban greening, and paper and wood production in northern China. However, given the high water use by the species and the regional dry environment, the long-term sustainability of these plantations needs to be evaluated. Currently, the understanding of energy partitioning and canopy resistance to water vapor and CO2 in poplar plantations is limited, impeding an accurate assessment of their true ecosystem functions. This study examined the variability of canopy bulk resistance parameters and energy partitioning over a four-year period encompassing both dry and wet conditions in a poplar (Populus euramericana CV. "74 / 76") plantation ecosystem located in northern China. Available energy (Net radiation Rn minus Soil Heat Flux, G) partitioning to latent (LE) and sensible (H) heat was responsive to climatological drought, with LE/(Rn-G) ranging from 62% in wet years (e.g. 2007 and 2008) to 53% in dry years (e.g. 2006 and 2009), and H/(Rn-G) from 25 to 33% between wet and dry years. Correspondingly, the Bowen ratio (β=H/LE) were 0.83 and 1.57. Surface resistance (Rs) had the greatest response to drought (+43%), but the aerodynamic and climatological resistances did not change significantly (p > 0.05). Partial correlation analysis indicated that Rs was the dominant factor in controlling the Bowen ratio. Furthermore, Rs was the major factor controlling LE during the growing season, even in wet years, as indicated by the decoupling coefficient (Ω = 0.45 and 0.39 in wet and dry years, respectively), and the LE / LEeq ratio ranged from 0.81 and 0.68 in wet and dry years, respectively. In general, the dry surface conditions dominated in this poplar plantation ecosystem regardless of soil water availability suggesting that fast-growing and water use-intensive species like poplar plantations are poorly adapted for the water limited region.

  19. Responses of surface ozone air quality to anthropogenic nitrogen deposition in the Northern Hemisphere

    Science.gov (United States)

    Zhao, Yuanhong; Zhang, Lin; Tai, Amos P. K.; Chen, Youfan; Pan, Yuepeng

    2017-08-01

    Human activities have substantially increased atmospheric deposition of reactive nitrogen to the Earth's surface, inducing unintentional effects on ecosystems with complex environmental and climate consequences. One consequence remaining unexplored is how surface air quality might respond to the enhanced nitrogen deposition through surface-atmosphere exchange. Here we combine a chemical transport model (GEOS-Chem) and a global land model (Community Land Model, CLM) to address this issue with a focus on ozone pollution in the Northern Hemisphere. We consider three processes that are important for surface ozone and can be perturbed by the addition of atmospheric deposited nitrogen - namely, emissions of biogenic volatile organic compounds (VOCs), ozone dry deposition, and soil nitrogen oxide (NOx) emissions. We find that present-day anthropogenic nitrogen deposition (65 Tg N a-1 to the land), through enhancing plant growth (represented as increases in vegetation leaf area index, LAI, in the model), could increase surface ozone from increased biogenic VOC emissions (e.g., a 6.6 Tg increase in isoprene emission), but it could also decrease ozone due to higher ozone dry deposition velocities (up to 0.02-0.04 cm s-1 increases). Meanwhile, deposited anthropogenic nitrogen to soil enhances soil NOx emissions. The overall effect on summer mean surface ozone concentrations shows general increases over the globe (up to 1.5-2.3 ppbv over the western US and South Asia), except for some regions with high anthropogenic NOx emissions (0.5-1.0 ppbv decreases over the eastern US, western Europe, and North China). We compare the surface ozone changes with those driven by the past 20-year climate and historical land use changes. We find that the impacts from anthropogenic nitrogen deposition can be comparable to the climate- and land-use-driven surface ozone changes at regional scales and partly offset the surface ozone reductions due to land use changes reported in previous studies

  20. Effects of shrub and tree cover increase on the near-surface atmosphere in northern Fennoscandia

    Science.gov (United States)

    Rydsaa, Johanne H.; Stordal, Frode; Bryn, Anders; Tallaksen, Lena M.

    2017-09-01

    Increased shrub and tree cover in high latitudes is a widely observed response to climate change that can lead to positive feedbacks to the regional climate. In this study we evaluate the sensitivity of the near-surface atmosphere to a potential increase in shrub and tree cover in the northern Fennoscandia region. We have applied the Weather Research and Forecasting (WRF) model with the Noah-UA land surface module in evaluating biophysical effects of increased shrub cover on the near-surface atmosphere at a fine resolution (5.4 km × 5.4 km). Perturbation experiments are performed in which we prescribe a gradual increase in taller vegetation in the alpine shrub and tree cover according to empirically established bioclimatic zones within the study region. We focus on the spring and summer atmospheric response. To evaluate the sensitivity of the atmospheric response to inter-annual variability in climate, simulations were conducted for two contrasting years, one warm and one cold. We find that shrub and tree cover increase leads to a general increase in near-surface temperatures, with the highest influence seen during the snowmelt season and a more moderate effect during summer. We find that the warming effect is stronger in taller vegetation types, with more complex canopies leading to decreases in the surface albedo. Counteracting effects include increased evapotranspiration, which can lead to increased cloud cover, precipitation, and snow cover. We find that the strength of the atmospheric feedback is sensitive to snow cover variations and to a lesser extent to summer temperatures. Our results show that the positive feedback to high-latitude warming induced by increased shrub and tree cover is a robust feature across inter-annual differences in meteorological conditions and will likely play an important role in land-atmosphere feedback processes in the future.

  1. Assessing Surface Hydrological Processes on a Rehabilitated Mine Landform in Northern Australia

    Science.gov (United States)

    Shao, Qi; Saynor, Mike; Lowry, John; Lu, Ping; Baumgartl, Thomas

    2015-04-01

    To assist with the evaluation of the proposed rehabilitation designs for the mine closure at a Uranium Mine, Northern Territory, Australia, a trial landform (200 m × 400 m) was constructed using waste rocks, with its surface ripped along the contour. The surface hydrological performance of this landform was investigated in this study. Field infiltration measurements were conducted using both large and regular ring infiltrometers to investigate the surface hydraulic properties, and water contents and surface runoff were monitored continuously in two 30 m × 30 m runoff plots for four years. A Cellular Automata based runoff model (RunCA) was also used to simulate the runoff behaviors under different rainfall conditions. Results showed a higher infiltration capacity in the areas of rip lines than the non-ripped areas due to the disturbance to the surface. Runoff coefficient was less than 6% and 10% for 80% of the 304 observed rainfall events on plot 1 and plot 2, respectively. The low levels of runoff were well explained by the simulated flow maps of RunCA, which demonstrated the roles of the rip lines in storing flow water and discontinuing the runoff paths. However, when the maximum storage capacity of these rip lines was exceeded during several large rainfall events, the runoff became much more significant and led to high potentials for erosion and landform instability. RunCA simulations on the virtual landforms with higher rip lines indicated dramatically reduced runoff rates. Therefore, it is suggested here that the current landform may be subjected to great runoff and erosion risks under extreme rainfall events, and raising the rip line height may potentially solve this problem.

  2. Overlap between Atlantic bluefin tuna spawning grounds and observed Deepwater Horizon surface oil in the northern Gulf of Mexico.

    Science.gov (United States)

    Muhling, B A; Roffer, M A; Lamkin, J T; Ingram, G W; Upton, M A; Gawlikowski, G; Muller-Karger, F; Habtes, S; Richards, W J

    2012-04-01

    The 2010 Deepwater Horizon oil spill impacted the northern Gulf of Mexico (GOM) during the spring spawning season of Atlantic bluefin tuna (BFT). Overlap between BFT spawning habitat and surface oil in the northern GOM was examined using satellite-derived estimates of oil coverage, and spawning habitat models. Results suggested that although eggs and larvae were likely impacted by oil-contaminated waters in the eastern GOM, high abundances of larvae were located elsewhere, especially in the western GOM. Overall, less than 10% of BFT spawning habitat was predicted to have been covered by surface oil, and less than 12% of larval BFT were predicted to have been located within contaminated waters in the northern GOM, on a weekly basis. Our results provide preliminary but important initial estimates of the effects of the spill on larval BFT mortality, as concern continues over the appropriate management responses to impacts of the spill. Copyright © 2012 Elsevier Ltd. All rights reserved.

  3. Natural oil slicks fuel surface water microbial activities in the northern Gulf of Mexico.

    Science.gov (United States)

    Ziervogel, Kai; D'Souza, Nigel; Sweet, Julia; Yan, Beizhan; Passow, Uta

    2014-01-01

    We conducted a series of roller tank incubations with surface seawater from the Green Canyon oil reservoir, northern Gulf of Mexico, amended with either a natural oil slick (GCS-oil) or pristine oil. The goal was to test whether bacterial activities of natural surface water communities facilitate the formation of oil-rich marine snow (oil snow). Although oil snow did not form during any of our experiments, we found specific bacterial metabolic responses to the addition of GCS-oil that profoundly affected carbon cycling within our 4-days incubations. Peptidase and β-glucosidase activities indicative of bacterial enzymatic hydrolysis of peptides and carbohydrates, respectively, were suppressed upon the addition of GCS-oil relative to the non-oil treatment, suggesting that ascending oil and gas initially inhibits bacterial metabolism in surface water. Biodegradation of physically dispersed GCS-oil components, indicated by the degradation of lower molecular weight n-alkanes as well as the rapid transformation of particulate oil-carbon (C: N >40) into the DOC pool, led to the production of carbohydrate- and peptide-rich degradation byproducts and bacterial metabolites such as transparent exopolymer particles (TEP). TEP formation was highest at day 4 in the presence of GCS-oil; in contrast, TEP levels in the non-oil treatment already peaked at day 2. Cell-specific enzymatic activities closely followed TEP concentrations in the presence and absence of GCS-oil. These results demonstrate that the formation of oil slicks and activities of oil-degrading bacteria result in a temporal offset of microbial cycling of organic matter, affecting food web interactions and carbon cycling in surface waters over cold seeps.

  4. Natural oil slicks fuel surface water microbial activities in the northern Gulf of Mexico

    Directory of Open Access Journals (Sweden)

    Kai eZiervogel

    2014-05-01

    Full Text Available We conducted a series of roller tank incubations with surface seawater from the Green Canyon oil reservoir, northern Gulf of Mexico, amended with either a natural oil slick (GCS-oil or pristine oil. The goal was to test whether bacterial activities of natural surface water communities facilitate the formation of oil-rich marine snow (oil snow. Although oil snow did not form during any of our experiments, we found specific bacterial metabolic responses to the addition of GCS-oil that profoundly affected carbon cycling within our 4-days incubations. Peptidase and -glucosidase activities indicative of bacterial enzymatic hydrolysis of peptides and carbohydrates, respectively, were suppressed upon the addition of GCS-oil relative to the non-oil treatment, suggesting that ascending oil and gas initially inhibits bacterial metabolism in surface water. Biodegradation of physically dispersed GCS-oil components indicated by the degradation of lower molecular weight n-alkanes as well as the rapid transformation of particulate oil-carbon (C: N >40 into the DOC pool, led to the production of carbohydrate- and peptide-rich degradation byproducts and bacterial metabolites such as transparent exopolymer particles (TEP. TEP formation was highest at day 4 in the presence of GCS-oil; in contrast, TEP levels in the non-oil treatment already peaked at day 2. Cell-specific enzymatic activities closely followed TEP concentrations in the presence and absence of GCS-oil. These results demonstrate that the formation of oil slicks and activities of oil-degrading bacteria result in a temporal offset of microbial cycling of organic matter, affecting food web interactions and carbon cycling in surface waters over cold seeps.

  5. Spatial diversity of bacterioplankton communities in surface water of northern South China Sea.

    Directory of Open Access Journals (Sweden)

    Jialin Li

    Full Text Available The South China Sea is one of the largest marginal seas, with relatively frequent passage of eddies and featuring distinct spatial variation in the western tropical Pacific Ocean. Here, we report a phylogenetic study of bacterial community structures in surface seawater of the northern South China Sea (nSCS. Samples collected from 31 sites across large environmental gradients were used to construct clone libraries and yielded 2,443 sequences grouped into 170 OTUs. Phylogenetic analysis revealed 23 bacterial classes with major components α-, β- and γ-Proteobacteria, as well as Cyanobacteria. At class and genus taxon levels, community structure of coastal waters was distinctively different from that of deep-sea waters and displayed a higher diversity index. Redundancy analyses revealed that bacterial community structures displayed a significant correlation with the water depth of individual sampling sites. Members of α-Proteobacteria were the principal component contributing to the differences of the clone libraries. Furthermore, the bacterial communities exhibited heterogeneity within zones of upwelling and anticyclonic eddies. Our results suggested that surface bacterial communities in nSCS had two-level patterns of spatial distribution structured by ecological types (coastal VS. oceanic zones and mesoscale physical processes, and also provided evidence for bacterial phylogenetic phyla shaped by ecological preferences.

  6. A review of surface heat-flow data of the northern Middle Atlas (Morocco)

    Science.gov (United States)

    Chiozzi, Paolo; Barkaoui, Alae-Eddine; Rimi, Abdelkrim; Verdoya, Massimo; Zarhloule, Yassine

    2017-12-01

    We revised thermal data available from water and oil wells in the northern sector of the Middle Atlas region. To avoid biased estimation of surface heat flow caused by advection likely occurring in shallow aquifers, temperature measurements in water boreholes were carefully inspected and selected. The heat flow in the oil wells was inferred by taking into account the porosity variation with depth, the temperature effect on thermal conductivity of the matrix and the pore fluid, together with the contribution of the radiogenic heat production. Moreover, the possible bias in heat flow caused by convection occurring in confined carbonate aquifers was evaluated. The results of heat flow slightly modify the picture reported in previous investigations. The heat flow value over the investigated region is rather uniform (about 80 mW m-2) and is similar in oil wells and in water boreholes. Geothermal calculations indicate that such a surface heat flow is compatible with a ∼70 km thick thermal lithosphere and normal thermal conditions in the asthenospheric mantle.

  7. Northern Korean Peninsula 1-D velocity model from surface wave dispersion and full-waveform data

    Science.gov (United States)

    Lee, S. J.; Rhie, J.; Kim, S.; Kang, T. S.; Cho, C.

    2016-12-01

    Monitoring seismic activities in the northern Korean Peninsula is important not only for understanding the characteristics of earthquakes but also for watching nuclear tests. To better monitor those natural and man-made seismic activities, reliable seismic velocity models are required. However, the seismic velocity structure of the region is not known well due to the lack of available seismic data directly measured in the region. This study presents 1-D velocity models of the region using two different datasets comprised of two-year-long continuous waveform and the 2013 North Korea nuclear test event waveform recorded at stations surrounding the region. Two reference 1-D models for the inland and offshore areas (Western East Sea) were estimated by 1-D inversion of surface wave dispersion measurements from ambient noise cross-correlations of the continuous waveform. To investigate the variations in the velocity models, many 1-D models for the paths between the 2013 nuclear test site and stations in China and South Korea were constructed by forward waveform modeling. The velocity variations are not significant for both models representing the inland and offshore paths, respectively. The 1-D models for the inland paths are similar to the models constructed for the southern Korean Peninsula. Interestingly, waveforms sampling through the offshore paths are not well explained by simple 1-D isotropic models. The preliminary result indicates that there exists radial anisotropy with SH being faster than SV by 3-5% in the upper mantle beneath the offshore northern Korean Peninsula, although further studies are necessary to explain the origin of anisotropy. A proper characterization of propagation effects along the offshore paths would be useful for monitoring future nuclear tests because many seismic stations in the eastern South Korea record waveforms sampling the offshore region from the nuclear test site to those stations.

  8. Energy partitioning and surface resistance of a poplar plantation in northern China

    Science.gov (United States)

    Kang, M.; Zhang, Z.; Noormets, A.; Fang, X.; Zha, T.; Zhou, J.; Sun, G.; McNulty, S. G.; Chen, J.

    2015-07-01

    Poplar (Populus sp.) plantations have been, on the one hand, broadly used in northern China for urban greening, combating desertification, as well as for paper and wood production. On the other hand, such plantations have been questioned occasionally for their possible negative impacts on water availability due to the higher water-use nature of poplar trees compared with other tree species in water-limited dryland regions. To further understand the acclimation of poplar species to semiarid environments and to evaluate the potential impacts of these plantations on the broader context of the region's water supply, we examine the variability of bulk resistance parameters and energy partitioning in a poplar (Populus euramericana cv. "74/76") plantation located in northern China over a 4-year period, encompassing both dry and wet conditions. The partitioning of available energy to latent heat flux (LE) decreased from 0.62 to 0.53 under mediated meteorological drought by irrigation applications. A concomitant increase in sensible heat flux (H) resulted in the increase of a Bowen ratio from 0.83 to 1.57. Partial correlation analysis indicated that surface resistance (Rs) normalized by leaf area index (LAI; Rs:LAI) increased by 50 % under drought conditions and was the dominant factor controlling the Bowen ratio. Furthermore, Rs was the main factor controlling LE during the growing season, even in wet years, as indicated by the decoupling coefficient (Ω = 0.45 and 0.39 in wet and dry years, respectively). Rs was also a major regulator of the LE / LEeq ratio, which decreased from 0.81 in wet years to 0.68 in dry years. All physiological and bioclimatological metrics indicated that the water demands of the poplar plantation were greater than the amount available through precipitation, highlighting the poor match of a water-intensive species like poplar for this water-limited region.

  9. Solar Cycle and Anthropogenic Forcing of Surface-Air Temperature at Armagh Observatory, Northern Ireland

    Science.gov (United States)

    Wilson, Robert M.

    2010-01-01

    A comparison of 10-yr moving average (yma) values of Armagh Observatory (Northern Ireland) surface-air temperatures with selected solar cycle indices (sunspot number (SSN) and the Aa geomagnetic index (Aa)), sea-surface temperatures in the Nino 3.4 region, and Mauna Loa carbon dioxide (CO2) (MLCO2) atmospheric concentration measurements reveals a strong correlation (r = 0.686) between the Armagh temperatures and Aa, especially, prior to about 1980 (r = 0.762 over the interval of 1873-1980). For the more recent interval 1963-2003, the strongest correlation (r = 0.877) is between Armagh temperatures and MLCO2 measurements. A bivariate fit using both Aa and Mauna Loa values results in a very strong fit (r = 0.948) for the interval 1963-2003, and a trivariate fit using Aa, SSN, and Mauna Loa values results in a slightly stronger fit (r = 0.952). Atmospheric CO2 concentration now appears to be the stronger driver of Armagh surface-air temperatures. An increase of 2 C above the long-term mean (9.2 C) at Armagh seems inevitable unless unabated increases in anthropogenic atmospheric gases can be curtailed. The present growth in 10-yma Armagh temperatures is about 0.05 C per yr since 1982. The present growth in MLCO2 is about 0.002 ppmv, based on an exponential fit using 10-yma values, although the growth appears to be steepening, thus, increasing the likelihood of deleterious effects attributed to global warming.

  10. Observational estimation of radiative feedback to surface air temperature over Northern High Latitudes

    Science.gov (United States)

    Hwang, Jiwon; Choi, Yong-Sang; Kim, WonMoo; Su, Hui; Jiang, Jonathan H.

    2018-01-01

    The high-latitude climate system contains complicated, but largely veiled physical feedback processes. Climate predictions remain uncertain, especially for the Northern High Latitudes (NHL; north of 60°N), and observational constraint on climate modeling is vital. This study estimates local radiative feedbacks for NHL based on the CERES/Terra satellite observations during March 2000-November 2014. The local shortwave (SW) and longwave (LW) radiative feedback parameters are calculated from linear regression of radiative fluxes at the top of the atmosphere on surface air temperatures. These parameters are estimated by the de-seasonalization and 12-month moving average of the radiative fluxes over NHL. The estimated magnitudes of the SW and the LW radiative feedbacks in NHL are 1.88 ± 0.73 and 2.38 ± 0.59 W m-2 K-1, respectively. The parameters are further decomposed into individual feedback components associated with surface albedo, water vapor, lapse rate, and clouds, as a product of the change in climate variables from ERA-Interim reanalysis estimates and their pre-calculated radiative kernels. The results reveal the significant role of clouds in reducing the surface albedo feedback (1.13 ± 0.44 W m-2 K-1 in the cloud-free condition, and 0.49 ± 0.30 W m-2 K-1 in the all-sky condition), while the lapse rate feedback is predominant in LW radiation (1.33 ± 0.18 W m-2 K-1). However, a large portion of the local SW and LW radiative feedbacks were not simply explained by the sum of these individual feedbacks.

  11. Surface Albedo Darkening from wildfires in Northern Sub-Saharan Africa

    Science.gov (United States)

    Gatebe, C. K.; Ichoku, C. M.; Poudal, R.; Roman, M. O.; Wilcox, E.

    2014-01-01

    Wildfires are recognized as a key physical disturbance of terrestrial ecosystems and a major source of atmospheric trace gases and aerosols. They are known to produce changes in landscape patterns and lead to changes in surface albedo that can persist for long periods. Here, we estimate the darkening of surface albedo due to wildfires in different land cover ecosystems in the Northern Sub-Saharan Africa using data from the Moderate Resolution Imaging Spectroradiometer (MODIS). We determined a decrease in albedo after fires over most land cover types (e.g. woody savannas: (-0.00352 0.00003) and savannas: (- 0.003910.00003), which together accounted for >86% of the total MODIS fire count between 2003 and 2011). Grasslands had a higher value (-0.00454 0.00003) than the savannas, but accounted for only about 5% of the total fire count. A few other land cover types (e.g. Deciduous broad leaf: (0.00062 0.00015), and barren: 0.00027 0.00019), showed an increase in albedo after fires, but accounted for less than 1% of the total fires. Albedo change due to wildfires is more important during the fire season (October-February). The albedo recovery progresses rapidly during the first year after fires, where savannas show the greatest recovery (>77%) within one year, while deciduous broadleaf, permanent wetlands and barren lands show the least one-year recovery (56%). The persistence of surface albedo darkening in most land cover types is limited to about six to seven years, after which at least 98% of the burnt pixels recover to their pre-fire albedo.

  12. Surface expression of intraplate postglacial faults in Sweden: from LiDAR data

    Science.gov (United States)

    Abduljabbar, Mawaheb; Ask, Maria; Bauer, Tobias; Lund, Björn; Smith, Colby; Mikko, Henrik; Munier, Raymond

    2016-04-01

    Large intraplate earthquakes, up to magnitude 8.0±0.3 (Lindblom et al. 2015) are inferred to have occurred in northern Fennoscandia at the end of, or just after the Weichselian deglaciation. More than a dozen large so-called postglacial faults (PGF) have been found in the region. The present-day microseismic activity is rather high in north Sweden, and there is a correlation between microseismicity and mapped PGF scarps: 71% of the observed earthquakes north of 66°N locate within 30 km to the southeast and 10 km to the northwest of PGFs (Lindblom et al., 2015). Surface expressions of PGFs in Sweden have mainly been mapped using aerial photogrammetry and trenching (e.g. Lagerbäck & Sundh 2008). Their detailed surface geometry may be investigated using the new high-resolution elevation model of Sweden (NNH) that has a vertical- and lateral resolution of 2 m and 0.25 m, respectively. With NNH data, known PGFs have been modified, and a number of new potential PGFs have been identified (Smith et al. 2014; Mikko et al. 2015). However, the detailed variation of their surface expression remains to be determined. Our main objective is to constrain the strike and surface offset (i.e., apparent vertical throw because of soil cover overlays the bedrock) across the PGF scarps. We anticipate using the results to constrain direction of fault motion and paleomagnitudes of PGFs, and in numerical analyzes to investigate the nature of PGFs. We have developed a methodology for analyzing PGF-geomorphology from LiDAR data using two main software platforms (Ask et al. 2015): (1) Move2015 by Midland Valley has been used for constructing 3D models of the surface traces of the PGFs to determine apparent vertical throw. The apparent hanging- and footwall cut off lines are digitized, and subsequent computation of coordinates is rather time efficient and provide continuous data of fault and soil geomorphology that can be statistically analyzed; and (2) ArcGIS 10.3 by Esri has mostly been

  13. Surface-expressed enolases of Plasmodium and other pathogens

    Directory of Open Access Journals (Sweden)

    Anil Kumar Ghosh

    2011-08-01

    Full Text Available Enolase is the eighth enzyme in the glycolytic pathway, a reaction that generates ATP from phosphoenol pyruvate in cytosolic compartments. Enolase is essential, especially for organisms devoid of the Krebs cycle that depend solely on glycolysis for energy. Interestingly, enolase appears to serve a separate function in some organisms, in that it is also exported to the cell surface via a poorly understood mechanism. In these organisms, surface enolase assists in the invasion of their host cells by binding plasminogen, an abundant plasma protease precursor. Binding is mediated by the interaction between a lysine motif of enolase with Kringle domains of plasminogen. The bound plasminogen is then cleaved by specific proteases to generate active plasmin. Plasmin is a potent serine protease that is thought to function in the degradation of the extracellular matrix surrounding the targeted host cell, thereby facilitating pathogen invasion. Recent work revealed that the malaria parasite Plasmodium also expresses surface enolase, and that this feature may be essential for completion of its life cycle. The therapeutic potential of targeting surface enolases of pathogens is discussed.

  14. Surface Water-Ice Deposits in the Northern Shadowed Regions of Ceres

    Science.gov (United States)

    Platz, T.; Nathues, A.; Schorghofer, N.; Preusker, F.; Mazarico, E.; Schroeder, S. E.; Byrne, S.; Kneissl, T.; Schmedemann, N.; Combe, J.-P.; hide

    2016-01-01

    Ceres, a dwarf planet located in the main asteroid belt, has a low bulk density, and models predict that a substantial amount of water ice is present in its mantle and outer shell. The Herschel telescope and the Dawn spacecraft have observed the release of water vapor from Ceres, and exposed water ice has been detected by Dawn on its surface at mid-latitudes. Water molecules from endogenic and exogenic sources can also be cold-trapped in permanent shadows at high latitudes, as happens on the Moon and Mercury. Here we present the first image-based survey of Ceres's northern permanent shadows and report the discovery of bright deposits in cold traps. We identify a minimum of 634 permanently shadowed craters. Bright deposits are detected on the floors of just 10 of these craters in multiscattered light. We spectroscopically identify one of the bright deposits as water ice. This detection strengthens the evidence that permanently shadowed areas have preserved water ice on airless planetary bodies.

  15. Analysis of overdeepened valleys using the digital elevation model of the bedrock surface of Northern Switzerland

    Energy Technology Data Exchange (ETDEWEB)

    Jordan, P.

    2010-11-15

    Based on surface and borehole information, together with pre-existing regional and local interpretations, a 7,150 square kilometre Raster Digital Elevation Model (DEM) of the bedrock surface of northern Switzerland was constructed using a 25 m cell size. This model represents a further important step in the understanding of Quaternary sediment distribution and is open to a broad field of application and analysis, including hydrogeological, geotechnical and geophysical studies as well as research in the field of Pleistocene landscape evolution. An analysis of the overdeepened valleys in the whole model area and, more specifically in the Reuss area, shows that, in most cases, overdeepening is restricted to the areas covered by the Last Glaciation Maximum (LGM). However, at various locations relatively narrow overdeepened valleys outreach the tongue basins and the LGM ice shield limits. Therefore, an earlier and further-reaching glacial event has probably contributed significantly to the overdeepening of these valleys. No significant overdeepening has been identified downstream of Boettstein (Aare) and Kaiserstuhl (Rhine), although the ice extended considerably further downstream, at least during the most extensive glaciation. Except for the bedrock between Brugg and Boettstein, no overdeepened valleys are found significantly north of the outcrop of Mesozoic limestone of the Folded and Tabular Jura. A detailed analysis of the Reuss area shows that the Lake and Suhre valleys are separated from the Emmen-Gisikon Reuss valley basin by a significant bedrock barrier. The individual bedrock valleys are divided into several sub-basins, indicating a multiphase evolution of the valleys. Some of the swells or barriers separating the sub-basins coincide with known late LGM retreat stages. In the Suhre valley, an old fluvial valley floor with restricted overdeepened sections is documented. (author)

  16. Modified expression of surface glyconjugates in stored human platelets

    International Nuclear Information System (INIS)

    Dhar, A.; Ganguly, P.

    1987-01-01

    Platelets are anucleated cells which play an important part in blood coagulation and thrombosis. These cells may be stored in the blood bank for only 4/5 days. In order to improve the storage of platelets, it is essential to first understand the changes in these cells due to storage. In this work, human platelets were stored in autologous plasma at 4 0 or 22 0 and their surface changes were monitored with three lectins - wheat germ afflutinin (WGA), concanavalin A (Con A) and lentil lectin (LL). Blood was drawn from healthy donors and platelet rich plasma (PRP) was collected by slow speed centrifugation. Platelets stored at either temperature for different times showed increased sensitivity to agglutination by WGA after 34-48 hrs. Lectins, Con A and LL, which were not agglutinating to fresh platelets readily caused agglutination after 48-72 hrs. The platelets stored for 25 hrs or longer period were insensitive to thrombin but showed enhanced aggregation with WGA. Labelling of surface glycoconjugates of stored platelets with 3 H-boro-hydride revealed progressive loss of a glycoprotein of Mr 150,000 (GPIb infinity) together with the appearance of components of Mr 69,000; Mr 60,000; Mr 25,000. New high molecular weight glycoproteins were also detected only in stored platelets. The author studies clearly indicate that modification or altered expression of platelets surface glycoproteins may be one factor of storage related dysfunction of platelets

  17. Sea Surface Salinity and Ocean Color Observations in the Northern Gulf

    Science.gov (United States)

    Wesson, J. C.; Burrage, D. M.; Wang, D. W.; Howden, S. D.

    2012-04-01

    Airborne mapping of Sea Surface Salinity (SSS) has been performed using L-Band radiometers for over 15 years, and has been operationally practical for over a decade. Ocean scale L-band observations of SSS are now obtained by satellite. ESA's SMOS has been operational for over two years and NASA's Aquarius satellite, launched in Jun, 2011, for over 6 months. Aircraft SSS complements satellite measurements by measuring nearer to coasts and with finer (˜1 km) spatial resolution. Due to the large effective pixel size of the satellite L-Band SSS measurements(˜35-80km), SMOS measurements do not reach the coast. Land microwave brightness signal in a given pixel contaminates the measurement of sea surface brightness temperature. However, the high signal to noise ratio (salinity contrast of 7-15 psu over 10km in some cases) of the coastal salinity signal, due to large freshwater sources, may dominate land contamination effects, to allow closer than usual SMOS SSS observations of strong coastal salinity patterns. An additional method to estimate SSS near coasts is using ocean color. Very near to coasts, freshwater sources such as rivers are relatively rich in Colored Dissolved Organic Matter (CDOM). As freshwater mixes with saltwater, salinity increases and CDOM concentrations fall. For conservative mixing, there is an inverse linear relation between CDOM and salinity, allowing estimates of SSS based on CDOM. The airborne sensors we use during STARRS flights include 2 SeaWifs airborne simulator sensors, one upward looking and one downward looking, as well as digital cameras, which we have used to identify color fronts. These provide ocean color measurements in addition to the STARRS microwave SSS measurements. We present results from an airborne campaign in the northern Gulf of Mexico, June 2-13, 2011. We made four types of flights. 1) Underflights of SMOS tracks at times coincident with SMOS passes. 2) Zig-zag flights along the coast, between Texas and Mississippi. 3

  18. Frequency of infection by hepatitis B virus and its surface mutants in a northern Indian population.

    Science.gov (United States)

    Singh, Harjeet; Aggarwal, Rakesh; Singh, R L; Naik, S R; Naik, Sita

    2003-01-01

    The reported prevalence of hepatitis B virus (HBV) infection in the Indian general population varies from 2% to 11%. Epidemiological studies conducted so far have selection biases, since these included populations of defined age group, gender, social class, high-risk group, etc. The present study was designed to look for the molecular epidemiology of HBV infection in the rural and urban general populations in India. Sera obtained from healthy volunteers during college and social service camps from parts of northern India were tested for HBsAg and anti-HBc using enzyme immunoassays and for HBV DNA using polymerase chain reaction and Southern blot hybridization. The amplification products were cloned and sequenced, and nucleotide and deduced amino acid sequences of the surface and polymerase genes were analyzed for mutations. Of the 730 subjects (rural 543, urban 187), 15 (2.1%) tested positive for HBsAg and 143 (19.5%) for anti-HBc; 10 were positive for both. The overall HBV exposure rate in the population was 20.3% (148/730). The HBsAg carrier rate was similar in the urban and rural populations (1.5% and 2.3%; p=ns), and anti-HBc positivity was lower in the urban population (8.5% vs. 23.3%; ptransfusion was associated with markers of exposure to HBV (10.2% vs. 4.6%; p=0.01). Among the 220 representative samples tested for HBV DNA, 14 (6.4%) were positive; of these, only four were positive for HBsAg or anti-HBc. Sequencing of a 388-nt segment of the S-gene from three individuals (two adw and one ayw subtype) revealed four mutations. Two and three of these led to amino acid changes in the HBV surface and polymerase genes, respectively; alterations in known cytotoxic T cell epitopes of HBV surface and polymerase proteins were observed in one individual each. None had the G587A mutation, which is known to be associated with loss of the 'a' determinant of HBsAg. Our study shows a high frequency of exposure to HBV infection in the Indian general population; a proportion

  19. Environmental assessment of polycyclic aromatic hydrocarbons (PAHs) in surface sediments of the Santander Bay, Northern Spain.

    Science.gov (United States)

    Viguri, J; Verde, J; Irabien, A

    2002-07-01

    Samples of intertidal surface sediments (0-2 cm) were collected in 17 stations of the Santander Bay, Cantabric Sea, Northern Spain. The concentrations of polycyclic aromatic hydrocarbons (PAHs), 16, were analysed by HPLC and MS detection. Surface sediments show a good linear correlation among the parameters of the experimental organic matter evaluation, where total carbon (TC) and loss on ignition (LOI) are approximately 2.5 and 5 times total organic carbon (TOC). A wide range of TOC from 0.08% to 4.1%, and a broad distribution of the sum of sigma16PAHs, from 0.02 to 344.6 microg/g d.w., which can be correlated by an exponential equation to the TOC, has been identified. A qualitative relationship may be established between the industrial input along the rivers and the concentration of sigma6PAHs in the sediments of the estuaries: Boo estuary (8404-4631 microg/g OC), Solia-San Salvador estuaries (305-113 microg/g OC) and Cubas estuary (31-32 microg/g OC). This work shows a dramatic change in the spatial distribution in the concentration of PAHs of intertidal surface sediments. The left edge of the Bay has the main traffic around the city and the major source of PAHs is from combustion processes and estuarine inputs, leading to medium values of PAHs in the sediments; the right edge of the Bay has much lesser anthropogenic activities leading to lower values of PAHs in sediments. The distribution of individual PAHs in sediments varies widely depending on their structure and molecular weight; the 4-6 ring aromatics predominate in polluted sediments due to their higher persistence. The isomer ratio does not allow any clear identification of the PAHs origin. Environmental evaluation according to Dutch guidelines and consensus sediment quality guidelines based on ecotoxicological data leads to the same conclusion, sediments in the Santander Bay show a very different environmental quality depending on the spatial position from heavily polluted/medium effects to non

  20. Screening for contaminants of emerging concern in Northern Colorado Plateau Network waters: 2015 surface-water data

    Science.gov (United States)

    Weissinger, R; Battaglin, William A.; Bradley, Paul M.

    2016-01-01

    In 2015, as part of an on-going screening program for contaminants of emerging concern (CECs) in conjunction with the Environmental Protection Agency (EPA) Region 8, surface waters at 18 locations in or near seven national park units within the Northern Colorado Plateau Network (NCPN) were sampled for pesticides and pesticide degradation products, pharmaceuticals and personal care products, hormones, organic-wastewater-indictor chemicals, and nutrients. Most sites were sampled in spring (May or June) and fall (September).

  1. Seismic imaging of the upper mantle beneath the northern Central Andean Plateau: Implications for surface topography

    Science.gov (United States)

    Ward, K. M.; Zandt, G.; Beck, S. L.; Wagner, L. S.

    2015-12-01

    Extending over 1,800 km along the active South American Cordilleran margin, the Central Andean Plateau (CAP) as defined by the 3 km elevation contour is second only to the Tibetan Plateau in geographic extent. The uplift history of the 4 km high Plateau remains uncertain with paleoelevation studies along the CAP suggesting a complex, non-uniform uplift history. As part of the Central Andean Uplift and the Geodynamics of High Topography (CAUGHT) project, we use surface waves measured from ambient noise and two-plane wave tomography to image the S-wave velocity structure of the crust and upper mantle to investigate the upper mantle component of plateau uplift. We observe three main features in our S-wave velocity model including (1), a high velocity slab (2), a low velocity anomaly above the slab where the slab changes dip from near horizontal to a normal dip, and (3), a high-velocity feature in the mantle above the slab that extends along the length of the Altiplano from the base of the Moho to a depth of ~120 km with the highest velocities observed under Lake Titicaca. A strong spatial correlation exists between the lateral extent of this high-velocity feature beneath the Altiplano and the lower elevations of the Altiplano basin suggesting a potential relationship. Non-uniqueness in our seismic models preclude uniquely constraining this feature as an uppermost mantle feature bellow the Moho or as a connected eastward dipping feature extending up to 300 km in the mantle as seen in deeper mantle tomography studies. Determining if the high velocity feature represents a small lithospheric root or a delaminating lithospheric root extending ~300 km into the mantle requires more integration of observations, but either interpretation shows a strong geodynamic connection with the uppermost mantle and the current topography of the northern CAP.

  2. Are Titan's radial Labyrinth terrains surface expressions of large laccoliths?

    Science.gov (United States)

    Schurmeier, L.; Dombard, A. J.; Malaska, M.; Radebaugh, J.

    2017-12-01

    The Labyrinth terrain unit may be the one of the best examples of the surface expression of Titan's complicated history. They are characterized as highly eroded, dissected, and elevated plateaus and remnant ridges, with an assumed composition that is likely organic-rich based on radar emissivity. How these features accumulated organic-rich sediments and formed topographic highs by either locally uplifting or surviving pervasive regional deflation or erosion is an important question for understanding the history of Titan. There are several subsets of Labyrinth terrains, presumably with differing evolutionary histories and formation processes. We aim to explain the formation of a subset of Labyrinth terrain units informally referred to as "radial Labyrinth terrains." They are elevated and appear dome-like, circular in planform, have a strong radial dissection pattern, are bordered by Undifferentiated Plains units, and are found in the mid-latitudes. Based on their shape, clustering, and dimensions, we suggest that they may be the surface expression of large subsurface laccoliths. A recent study by Manga and Michaut (Icarus, 2017) explained Europa's lenticulae (pits, domes, spots) with the formation of saucer-shaped sills that form laccoliths around the brittle-ductile transition depth within the ice shell (1-5 km). Here, we apply the same scaling relationships and find that the larger size of radial labyrinth terrains with Titan's higher gravity implies deeper intrusion depths of around 20-40 km. This intrusion depth matches the expected brittle-ductile transition on Titan based on our finite element simulations and yield strength envelope analyses. We hypothesize that Titan's radial labyrinth terrains formed as cryovolcanic (water) intrusions that rose to the brittle-ductile transition within the ice shell where they spread horizontally, and uplifted the overlying ice. The organic-rich sedimentary cover also uplifted, becoming more susceptible to pluvial and fluvial

  3. Tracking Hydrothermal Fluid Pathways from Surface Alteration Mineralogy: The Case of Licancura Geothermal Field, Northern Chile

    Science.gov (United States)

    Camus, E.; Elizalde, J. D.; Morata, D.; Wechsler, C.

    2017-12-01

    In geothermal systems alteration minerals are evidence of hot fluid flow, being present even in absence of other surface manifestations. Because these minerals result from the interaction between geothermal fluids and surrounding host rocks, they will provide information about features of thermal fluids as temperature, composition and pH, allowing tracking their changes and evolution. In this work, we study the Licancura Geothermal field located in the Andean Cordillera in Northern Chile. The combination of Principal Components Analysis on ASTER-L1T imagery and X Ray Diffraction (XRD) allow us to interpret fluid conditions and the areas where fluid flow took place. Results from red, green, blue color composite imagery show the presence of three types of secondary paragenesis. The first one corresponds to hematite and goethite, mainly at the east of the area, in the zone of eroded Pliocene volcanic edifices. The second one, mainly at the center of the area, highlighting propylitic alteration, includes minerals such as chlorite, illite, calcite, zeolites, and epidote. The third paragenesis, spatially related to the intersection between faults, represents advanced argillic alteration, includes minerals as alunite, kaolinite, and jarosite. XRD analysis support results from remote sensing techniques. These results suggest an acid pH hydrothermal fluid reaching temperatures at surface up to 80-100°C, which used faults as a conduit, originating advanced argillic minerals. The same fluid was, probably, responsible for propylitic paragenesis. However, iron oxides paragenesis identified in the area of eroded volcanoes probably corresponds to other processes associated with weathering rather than geothermal activity. In this work, we propose the applicability of remote sensing techniques as a first level exploration tool useful for high-altitude geothermal fields. Detailed clay mineral studies (XRD and SEM) would allow us to a better characterization of the geothermal fluid

  4. The Leishmania promastigote surface antigen 2 complex is differentially expressed during the parasite life cycle.

    Science.gov (United States)

    Handman, E; Osborn, A H; Symons, F; van Driel, R; Cappai, R

    1995-11-01

    The promastigote surface antigen 2 (PSA-2) complex comprises a family of antigenically similar polypeptides of M(r) 96,000, 80,000 and 50,000, anchored to the membrane with glycosylphosphatidylinositol. Although PSA-2 was initially detected only in promastigotes, Northern blot analysis indicated that mRNA transcripts are also present in amastigotes. Unlike the situation in promastigotes, where at least four major transcripts (2.6-5.3 kb) were detected, only one major (2.6 kb) and two minor transcripts were present in amastigotes. A cDNA clone encoding a member of the PSA-2 family expressed in amastigotes was isolated using DNA probes. The predicted protein sequence of M(r) 40,000 is distinct from promastigote sequences, but shows significant similarity to previously described members of the family from L major and L amazonensis. Antibodies to the carboxyl terminal sequence conserved in all L major PSA-2 studied to date, as well as antibodies affinity purified on the amastigote cDNA-derived polypeptide recognized a major M(r) 50,000 amastigote polypeptide. Immuno-electron microscopy localized both promastigote and amastigote PSA-2 to the cell surface. The expression of PSA-2 polypeptides during the transformation of amastigotes into promastigotes was ordered in a time-dependent manner, with the promastigote M(r) 80000 polypeptide appearing first, followed by the M(r) 96000 polypeptide. In contrast to the glycosylphosphatidylinositol anchor of promastigote PSA-2, which could be hydrolysed by phosphatidylinositol-specific phospholipase C, the amastigote form was resistant to this enzyme.

  5. Vesicular Stomatitis Virus Infection Promotes Immune Evasion by Preventing NKG2D-Ligand Surface Expression

    DEFF Research Database (Denmark)

    Jensen, Helle; Andresen, Lars; Nielsen, Jens

    2011-01-01

    leads to a robust induction of MICA mRNA expression, however the subsequent surface expression is potently hindered. Thus, VSV lines up with human cytomegalovirus (HCMV) and adenovirus, which actively subvert the immune system by negatively affecting NKG2D-ligand surface expression. VSV infection caused...

  6. Habitat manipulation influences northern bobwhite resource selection on a reclaimed surface mine

    Science.gov (United States)

    Brooke, Jarred M.; Peters, David C.; Unger, Ashley M.; Tanner, Evan P.; Harper, Craig A.; Keyser, Patrick D.; Clark, Joseph D.; Morgan, John J.

    2015-01-01

    More than 600,000 ha of mine land have been reclaimed in the eastern United States, providing large contiguous tracts of early successional vegetation that can be managed for northern bobwhite (Colinus virginianus). However, habitat quality on reclaimed mine land can be limited by extensive coverage of non-native invasive species, which are commonly planted during reclamation. We used discrete-choice analysis to investigate bobwhite resource selection throughout the year on Peabody Wildlife Management Area, a 3,330-ha reclaimed surface mine in western Kentucky. We used a treatment-control design to study resource selection at 2 spatial scales to identify important aspects of mine land vegetation and whether resource selection differed between areas with habitat management (i.e., burning, disking, herbicide; treatment) and unmanaged units (control). Our objectives were to estimate bobwhite resource selection on reclaimed mine land and to estimate the influence of habitat management practices on resource selection. We used locations from 283 individuals during the breeding season (1 Apr–30 Sep) and 136 coveys during the non-breeding season (1 Oct–Mar 31) from August 2009 to March 2014. Individuals were located closer to shrub cover than would be expected at random throughout the year. During the breeding season, individuals on treatment units used areas with smaller contagion index values (i.e., greater interspersion) compared with individuals on control units. During the non-breeding season, birds selected areas with greater shrub-open edge density compared with random. At the microhabitat scale, individuals selected areas with increased visual obstruction >1 m aboveground. During the breeding season, birds were closer to disked areas (linear and non-linear) than would be expected at random. Individuals selected non-linear disked areas during winter but did not select linear disked areas (firebreaks) because they were planted to winter wheat each fall and

  7. Sea-surface salinity variations in the northern Caribbean Sea across the Mid-Pleistocene Transition

    Directory of Open Access Journals (Sweden)

    S. Sepulcre

    2011-02-01

    Full Text Available By reconstructing past hydrologic variations in the Northern Caribbean Sea and their influence on the stability of the Atlantic Meridional Overturning Circulation (AMOC during the last 940 ka, we seek to document climate changes in this tropical area in response to the Mid-Pleistocene Transition (MPT. Using core MD03-2628, we estimated past changes in sea surface salinity (SSS using Δδ18O, the difference between the modern, and the past δ18O of seawater (obtained by combining alkenone thermometer data with the δ18O of the planktonic foraminifera Globigerinoides rube (white and corrected for ice-sheet volume effects. Today, the lowest SSS values in the area studied are associated with the northernmost location of the Inter-Tropical Convergence Zone (ITCZ. The Δδ18O record obtained from core MD03-2628 exhibits glacial/interglacial cyclicity with higher values during all glacial periods spanning the last 940 ka, indicating increased SSS. A long-term trend was also observed in the Δδ18O values that exhibited a shift toward lower values for interglacial periods during the last 450 ka, as compared to interglacial stages older than 650 ka. A rise in SSS during glacial stages may be related to the southernmost location of the ITCZ, which is induced by a steeper cross-equator temperature gradient and associated with reduced northward cross-equatorial oceanic transport. Therefore, the results suggest a permanent link between the tropical salinity budget and the AMOC during the last 940 ka. Following the MPT, lower salinities during the last five interglacial stages indicated a northernmost ITCZ location that was forced by changes in the cross-equator temperature gradient and that was associated with the poleward position of Southern Oceanic Fronts that amplify the transport of heat and moisture to the North Atlantic. These processes may have contributed to the amplification of the

  8. Energy partitioning and surface resistance of a poplar plantation in northern China

    Science.gov (United States)

    M. Kang; Z. Zhang; A. Noormets; X. Fang; T. Zha; J. Zhou; G. Sun; S. G. McNulty; J. Chen

    2015-01-01

    Poplar (Populus sp.) plantations have been, on the one hand, broadly used in northern China for urban greening, combating desertification, as well as for paper and wood production. On the other hand, such plantations have been questioned occasionally for their possible negative impacts on water availability due to the higher water-use nature of...

  9. Near 7-day response of ocean bottom pressure to atmospheric surface pressure and winds in the northern South China Sea

    Science.gov (United States)

    Zhang, Kun; Zhu, Xiao-Hua; Zhao, Ruixiang

    2018-02-01

    Ocean bottom pressures, observed by five pressure-recording inverted echo sounders (PIESs) from October 2012 to July 2014, exhibit strong near 7-day variability in the northern South China Sea (SCS) where long-term in situ bottom pressure observations are quite sparse. This variability was strongest in October 2013 during the near two years observation period. By joint analysis with European Center for Medium-Range Weather Forecasts (ECMWF) data, it is shown that the near 7-day ocean bottom pressure variability is closely related to the local atmospheric surface pressure and winds. Within a period band near 7 days, there are high coherences, exceeding 95% significance level, of observed ocean bottom pressure with local atmospheric surface pressure and with both zonal and meridional components of the wind. Ekman pumping/suction caused by the meridional component of the wind in particular, is suggested as one driving mechanism. A Kelvin wave response to the near 7-day oscillation would propagate down along the continental slope, observed at the Qui Nhon in the Vietnam. By multiple and partial coherence analyses, we find that local atmospheric surface pressure and Ekman pumping/suction show nearly equal influence on ocean bottom pressure variability at near 7-day periods. A schematic diagram representing an idealized model gives us a possible mechanism to explain the relationship between ocean bottom pressure and local atmospheric forcing at near 7-day periods in the northern SCS.

  10. Cell-surface expression of Hsp70 on hematopoietic cancer cells after inhibition of HDAC activity

    DEFF Research Database (Denmark)

    Jensen, Helle; Andresen, Lars; Hansen, Karen Aagaard

    or normal ER/Golgi transport did not affect Hsp70 surface expression. Intracellular Calcium and the transcription factor Sp1, that has previously been shown to be important for the intracellular stress mediated by HDAC-inhibitors, were not involved in Hsp70 surface expression. We also found that HDAC...

  11. Cell-surface expression of Hsp70 on hematopoietic cancer cells after inhibition of HDAC activity

    DEFF Research Database (Denmark)

    Jensen, Helle; Andresen, Lars; Hansen, Karen Aagaard

    clinically applied reagents, such as alkyl-lysophospholipides, chemotherapeutic agents, and anti-inflammatory reagents, have been found to enhance Hsp70 surface expression on cancer cells. We have found that inhibition of histone deacetylase (HDAC) activity leads to surface expression of Hsp70 on various...

  12. Exploring the relationship between meteorology and surface PM2.5 in Northern India

    Science.gov (United States)

    Schnell, J.; Naik, V.; Horowitz, L. W.; Paulot, F.; Ginoux, P. A.

    2017-12-01

    Northern India is one of the most polluted and densely populated regions in world. Accurately modeling pollution in the region is difficult due to the extreme conditions with respect to emissions, meteorology, and topography, but it is paramount in order to understand how future changes in emissions and climate may alter the region's pollution regime. We evaluate a developmental version of the new-generation NOAA GFDL Atmospheric Model, version 4 (AM4) in its ability to simulate observed wintertime PM2.5 and its relationship to meteorology over the Northern India (23°N-31°N, 68°E-90°E). We perform two simulations of the GFDL-AM4 nudged to observed meteorology for the period (1980-2016) with two emission inventories developed for CMIP5 and CMIP6 and compare results with observations from India's Central Pollution Control Board (CPCB) for the period 1 October 2015 - 31 March 2016. Overall, our results indicate that the simulation with CMIP6 emissions has substantially reduced the low model bias in the region. The AM4, albeit biased low, generally simulates the magnitude and daily variability in observed total PM2.5. Ammonium nitrate and ammonium sulfate are the primary components of PM2.5 in the model, and although not directly observed, correlations of total observed PM2.5 and meteorology with the modeled individual PM2.5 components suggest the same for the observations. The model correctly reproduces the shape and magnitude of the seasonal cycle of PM2.5; but for the diurnal cycle, it misses the early evening rise and secondary maximum found in the observations. Observed PM2.5 abundances within the densely populated Indo-Gangetic Plain are by far the highest and are closely related to boundary layer meteorology, specifically relative humidity, wind speed, boundary layer height, and inversion strength. The GFDL-AM4 reproduces the observed pollution gradient over Northern India as well as the strength of the meteorology-PM2.5 relationship in most locations.

  13. Changes in gene expression linked with adult reproductive diapause in a northern malt fly species: a candidate gene microarray study

    Directory of Open Access Journals (Sweden)

    Hoikkala Anneli

    2010-02-01

    Full Text Available Abstract Background Insect diapause is an important biological process which involves many life-history parameters important for survival and reproductive fitness at both individual and population level. Drosophila montana, a species of D. virilis group, has a profound photoperiodic reproductive diapause that enables the adult flies to survive through the harsh winter conditions of high latitudes and altitudes. We created a custom-made microarray for D. montana with 101 genes known to affect traits important in diapause, photoperiodism, reproductive behaviour, circadian clock and stress tolerance in model Drosophila species. This array gave us a chance to filter out genes showing expression changes during photoperiodic reproductive diapause in a species adapted to live in northern latitudes with high seasonal changes in environmental conditions. Results Comparisons among diapausing, reproducing and young D. montana females revealed expression changes in 24 genes on microarray; for example in comparison between diapausing and reproducing females one gene (Drosophila cold acclimation gene, Dca showed up-regulation and 15 genes showed down-regulation in diapausing females. Down-regulation of seven of these genes was specific to diapause state while in five genes the expression changes were linked with the age of the females rather than with their reproductive status. Also, qRT-PCR experiments confirmed couch potato (cpo gene to be involved in diapause of D. montana. Conclusions A candidate gene microarray proved to offer a practical and cost-effective way to trace genes that are likely to play an important role in photoperiodic reproductive diapause and further in adaptation to seasonally varying environmental conditions. The present study revealed two genes, Dca and cpo, whose role in photoperiodic diapause in D. montana is worth of studying in more details. Also, further studies using the candidate gene microarray with more specific experimental

  14. Landscape and Seasonal Variability in CO2 Efflux from Soil and Water Surfaces in the Northern Pantanal

    Science.gov (United States)

    Couto, E. G.; Pinto-Jr, O. B.; Lathuilliere, M. J.; Dalmagro, H. J.; Johnson, M. S.

    2014-12-01

    The Pantanal is one of the largest wetlands in the world, with an area of 150,000 km2. It extends over three countries (Brazil, Paraguay and Bolivia) with 80% located in the Brazilian states of Mato Grosso and Mato Grosso do Sul. Ecosystems in the Northern Pantanal rely on a seasonal flood pulse in phase with the wet season, which inundates grasslands and forests between January and May. This pulse results in an important change in local biogeochemistry. Inundation saturates the Pantanal's soils with changes in the balance of carbon dioxide and other greenhouse gases. This study summarize the main CO2 effluxes measured from a variety of landscape in the Northern Pantanal during dry and wet seasons, fully or partially inundated soils, as part of Project 2.01 of the Brazilian Institute for Science and Technology in Wetlands (Monitoring aquatic carbon fluxes and water quality). Using a network of dataloggers installed in 4 environments, we have been modeling soil CO2 efflux on a half hourly basis through a combination of infrared gas analyzers measurements and laboratory soil physical parameter estimates. The selected environments presented unique biogeochemical behavior as they relate to inundation and soil type. So far, we have estimated average CO2 efflux in 3 environments with mean values of 3.53 µmol m-2 s-1 (soil CO2 efflux for the "Carrapatal" tree island), 3.41 µmol m-2 s-1 (soil CO2 efflux for the "Baia das Pedras" tree island), and 1.79 µmol m-2 s-1 (aquatic CO2 evasion from the water surface of the "Cambarazal" flooded forest). More measurements are currently underway to complete the landscape variability in CO2 effluxes in the Northern Pantanal.

  15. Evolution of Mars’ Northern Polar Seasonal CO2 deposits: variations in surface brightness and bulk density

    Science.gov (United States)

    Mount, Christopher P.; Titus, Timothy N.

    2015-01-01

    Small scale variations of seasonal ice are explored at different geomorphic units on the Northern Polar Seasonal Cap (NPSC). We use seasonal rock shadow measurements, combined with visible and thermal observations, to calculate density over time. The coupling of volume density and albedo allows us to determine the microphysical state of the seasonal CO2 ice. We find two distinct endmembers across the NPSC: 1) Snow deposits may anneal to form an overlying slab layer that fractures. These low density deposits maintain relatively constant densities over springtime. 2) Porous slab deposits likely anneal rapidly in early spring and fracture in late spring. These high density deposits dramatically increase in density over time. The endmembers appear to be correlated with latitude.

  16. Expressed Emotion and its Relationship to Adolescent Depression and Antisocial Behavior in Northern Taiwan

    OpenAIRE

    Bee-Horng Lue; Wen-Chi Wu; Lee-Lan Yen

    2010-01-01

    Despite widespread recognition of the occurrence of antisocial behavior and depression in adolescents, the specifics of the relationship between them have not been clarified. The purpose of this study was to investigate the role of expressed emotion as a proximal factor for depression and antisocial behavior among adolescents, by looking at direct and indirect relationships. Methods: Secondary data analysis using path analysis was carried out on 2004 data from the Child and Adolescent Beha...

  17. Spatial and temporal variability of N2O in the surface groundwater: a detailed analysis from a sandy aquifer in northern Germany

    OpenAIRE

    Heide, C.; Böttcher, J.; Deurer, M.; Duijnisveld, W.; Weymann, D.; Well, R.

    2009-01-01

    The knowledge of the spatial and temporal variability of N2O concentrations in surface groundwater is the first step towards upscaling of potential indirect N2O emissions from the scale of localized samples to aquifers. This study aimed to investigate the spatial and the temporal variability of N2O concentrations at different scales in the surface groundwater of a denitrifying aquifer in northern Germany. The spatial variability of N2O concentrations in the surface groundwater was analysed at...

  18. Cloud Properties Observed From the Surface and by Satellite at the Northern Edge of the Southern Ocean

    Science.gov (United States)

    Alexander, S. P.; Protat, A.

    2018-01-01

    A Raman depolarization lidar was deployed at Cape Grim, Australia (40.7°S, 144.7°E), at the northern edge of the Southern Ocean from July 2013 to February 2014 from which we determine cloud boundaries, cloud phase, ice virga, and cloud effective top heights. We compare surface-based lidar with results from the raDAR/liDAR (DARDAR) data set within 1,000 km of Cape Grim. DARDAR combines information from the CloudSat and Cloud-Aerosol Lidar with Orthogonal Polarization (CALIOP) instruments. We extract single-layer clouds that are sufficiently thin for signal to be present on the farside of the cloud and which have a liquid cloud top phase. These conditions maximize the likelihood that both surface-based lidar and DARDAR are observing the full vertical extent of the same clouds. Differences in low-level cloud occurrence frequencies for these single-layer clouds reveal that DARDAR underestimates cloud at 0.2-1.0 km altitude by a factor of 3 compared with the surface-based lidar. When multiple cloud decks are present, the underestimate in this altitude region is around 2.5 times. Heterogeneous glaciation observed by the Cape Grim lidar in midlevel stratiform supercooled water clouds is similar to that reported by previous surface-based observations adjacent to the Southern Ocean, with half of these clouds precipitating ice at cloud top temperatures of -20°C. This transition occurs around -15°C in the DARDAR data set, and this difference is likely due to the reduced sensitivity of surface-based lidar in detecting precipitating ice compared with what a surface-based radar could observe.

  19. Five Year Mean Surface Chlorophyll Estimates in the Northern Gulf of Mexico for 2005 through 2009

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — These images were created by combining the mean surface chlorophyll estimates to produce seasonal representations for winter, spring, summer and fall. Winter...

  20. Circulation and water characteristics on isanosteric surfaces in the Northern Arabian Sea during February-April

    Digital Repository Service at National Institute of Oceanography (India)

    Das, V.K.; Gouveia, A.D.; Varma, K.K.

    Distribution of water characteristics and circulation were studied on 260, 160, 140, 120 and 80 cl/t surfaces using isanosteric analysis and geostrophic computation The circulation pattern consisted of several eddies and meanders The most prominent...

  1. The surface expression of HLA-F on decidual trophoblasts increases from mid to term gestation.

    Science.gov (United States)

    Shobu, Takanori; Sageshima, Noriko; Tokui, Hiroshi; Omura, Motoko; Saito, Keigo; Nagatsuka, Yuka; Nakanishi, Mari; Hayashi, Yukio; Hatake, Katsuhiko; Ishitani, Akiko

    2006-12-01

    HLA-F has recently only begun to be studied in earnest, and has been thought not to be expressed on the cell surface. However, in our previous report, we demonstrated surface expression of HLA-F on extravillous trophoblasts (EVTs) invading the decidua in term placental tissues. To better understand its function, we attempted to determine when surface expression of HLA-F begins during normal pregnancy, and whether there is a difference in expression between normal and preeclamptic placentas, by comparing the expression of HLA-G and -E by immunohistochemical staining with anti-HLA-E, -F and -G antibodies (3D12, 3D11 and 87G, respectively). In EVTs, HLA-F was expressed only in the cytoplasm weakly during the first trimester, after which expression increased and moved to the cell surface with the progression of pregnancy from the second trimester, which was confirmed by the results of double-labeled immunofluorescence staining with anti-HLA-F and anti-HLA-G antibodies, and by flow cytometry using trophoblasts isolated from the decidua. HLA-E showed similar expression as HLA-F, though it was expressed on the cell surface from the first trimester, while HLA-G was expressed strongly in the cytoplasm and on the cell surface during all stages of pregnancy. The expressions of HLA-E, -F and -G in preeclamptic placentas were not different from those in normal placentas, though there were a greater number of necrotic EVTs in preeclampsia. The increase in expression of HLA-E and HLA-F from the second trimester to full term was coincident with the timing of rapid growth of the fetus. Our results suggest that these may function together to prepare an environment that supports fetal growth.

  2. Dynamic modeling of organophosphate pesticide load in surface water in the northern San Joaquin Valley watershed of California

    Science.gov (United States)

    Luo, Y.; Zhang, X.; Liu, X.; Ficklin, D. L.; Zhang, M.

    2008-12-01

    The hydrology, sediment, and pesticide transport components of the Soil and Water Assessment Tool (SWAT) were evaluated on the northern San Joaquin Valley watershed of California. The Nash Sutcliffe coefficients for monthly stream flow and sediment load ranged from 0.49 to 0.99 over the watershed during the study period of 1992 to 2005. The calibrated SWAT model was applied to simulate fate and transport processes of two organophosphate pesticides of diazinon and chlorpyrifos at watershed scale. The model generated satisfactory predictions of dissolved pesticide loads relative to the monitoring data. The model also showed great success in capturing spatial patterns of dissolved diazinon and chlorpyrifos loads according to the soil properties and landscape morphology over the large agricultural watershed. This study indicated that curve number was the major factor influencing the hydrology while pesticide fate and transport were mainly affected by surface runoff and pesticide application timing in the study area.

  3. Quantifying Surface Water Dynamics at 30 Meter Spatial Resolution in the North American High Northern Latitudes 1991-2011

    Science.gov (United States)

    Carroll, Mark; Wooten, Margaret; DiMiceli, Charlene; Sohlberg, Robert; Kelly, Maureen

    2016-01-01

    The availability of a dense time series of satellite observations at moderate (30 m) spatial resolution is enabling unprecedented opportunities for understanding ecosystems around the world. A time series of data from Landsat was used to generate a series of three maps at decadal time step to show how surface water has changed from 1991 to 2011 in the high northern latitudes of North America. Previous attempts to characterize the change in surface water in this region have been limited in either spatial or temporal resolution, or both. This series of maps was generated for the NASA Arctic and Boreal Vulnerability Experiment (ABoVE), which began in fall 2015. These maps show a nominal extent of surface water by using multiple observations to make a single map for each time step. This increases the confidence that any detected changes are related to climate or ecosystem changes not simply caused by short duration weather events such as flood or drought. The methods and comparison to other contemporary maps of the region are presented here. Initial verification results indicate 96% producer accuracy and 54% user accuracy when compared to 2-m resolution World View-2 data. All water bodies that were omitted were one Landsat pixel or smaller, hence below detection limits of the instrument.

  4. Transient Surface Deformation of Northern Taiwan, 2007-2011, Using Persistent Scattered InSAR with ALOS Data

    Science.gov (United States)

    Wang, C.; Chang, W.; Chang, C.

    2013-12-01

    The Taipei basin, triangular in shape and located in the northern Taiwan, is now developed into the most densely populated area and also the capital of politics and economics in Taiwan. North of the Taipei basin, the Tatun volcano group was proposed to be the cause of extensional collapse during the Pleistocene following the collision between the Luzon volcanic arc and the Eurasian continental margin at about 5 Ma. We investigated the contemporary surface deformation of the northern Taiwan using ALOS images that cover the Taipei basin and its surrounding mountainous area. The Differential Interferometric Synthetic Aperture Radar (DInSAR) technique has been widely used in the past ten years. However, the mountainous areas surrounding the basin are mostly covered with densely various vegetations that reduce signal-to-noise ratio in the interferograms. Therefore, the DInSAR technique is not effective for measuring the surface deformation in and around the Taipei basin, including the Tatun volcano area, and consequently the Persistent Scatterer (PS) and small baseline (SB) InSAR techniques have been employed to extract phase signals of the chosen PS points. In this study, we aim to measure the ground deformation of northern Taiwan by processing the spaceborne radar interferometry data of ALOS acquired from 2007 to 2011 using PSInSAR and SBInSAR techniques. Compared with the Envisat and ERS images used by previous studies, L-band PALSAR images can produce more PS points in the region covered by dense vegetation so that our results reveal a higher resolution of ground deformation. The mean Line of Sight (LOS) velocity field of up to 8 mm/yr in the central Tatun volcanic area, and up to 5 mm/yr in the Taipei basin with higher rate at the hanging wall of the Sanchiao fault than the footwall. (See the Figure.) While previous studies indicated that the Taipei basin had experienced ground uplift from 1993 to 2001 and subsidence from 2003 to 2008, our results show a return to

  5. Dissolved lipid production in the Northern Adriatic (Mediterranean) in response to sea surface warmin

    Science.gov (United States)

    Gasparovic, Blazenka; Novak, Tihana; Godrijan, Jelena; Mlakar, Marina; MAric, Daniela; Djakovac, Tamara

    2017-04-01

    Marine dissolved organic matter (OM) represents one of the largest active pools of organic carbon in the global carbon cycle. Oceans and seas are responsible for half of global primary production. Ocean warming caused by climate change is already starting to impact the marine life that necessary will have impact on ocean productivity. The partition of OM production by phytoplankton (major OM producer in seas and ocens) in the conditions of rising temperatures may considerably change. This has implications for the export of organic matter from the photic zone. In this study, we set out to see how annual temperature changes between 10 and 30 C in the Northern Adriatic (Mediterranean) affect production of DOM and particularly dissolved lipids and lipid classes. We have sampled at two stations being oligotrophic and mesotrophic where we expected different system reaction to temperature changes. In addition, we performed microcosm incubations covering temperature range of the NA with nutrient amendments to test whether changes in the available nutrients would reflect those of dissolved OM in the NA. We have selected to work with extracellular OM produced during growth of diatom Chaetoceros curvisetus cultures according to the criteria that genera Chaetoceros are important component of the phytoplankton in the NA and are often among bloom-forming taxa. Details on the dissolved lipid and lipid classes production as plankton responce to rising temperature will be discussed.

  6. Distribution of tetraether lipids in surface sediments of the northern South China Sea: Implications for TEX86 proxies

    Directory of Open Access Journals (Sweden)

    Huangmin Ge

    2013-03-01

    Full Text Available Archaea have unique glycerol dialkyl glycerol tetraether (GDGT lipids that can be used to develop paleotemperature proxies such as TEX86. This research is to validate proposed GDGT-proxies for paleotemperature determination in the South China Sea (SCS. Samples were collected from core-top sediments (0–5 cm in the northern SCS. Total lipids were extracted to obtain core GDGTs, which were identified and quantified using liquid chromatography-mass spectrometry (LC-MS. The abundance of isoprenoidal GDGTs (iGDGTs ranged from 271.5 ng/g dry sediment to 1266.3 ng/g dry sediment, whereas the branched GDGTs (bGDGTs, supposedly derived from terrestrial sources, ranged from 22.2 ng/g dry sediment to 56.7 ng/g dry sediment. The TEX86-derived sea surface temperatures ranged from 20.9 °C in the coast (water depth  1000 m. TEX86-derived temperatures near shore (<160 m water depth averaged 23.1 ± 2.5 °C (n = 4, which were close to the satellite-derived winter mean sea surface temperature (average 22.6 ± 1.0 °C, n = 4; whereas the TEX86-derived temperatures offshore averaged 27.4 ± 0.3 °C (n = 7 and were consistent with the satellite mean annual sea surface temperature (average 26.8 ± 0.4 °C, n = 7. These results suggest that TEX86 may record the sea surface mean annual temperature in the open ocean, while it likely records winter sea surface temperature in the shallower water.

  7. Vesicular stomatitis virus infection promotes immune evasion by preventing NKG2D-ligand surface expression.

    Science.gov (United States)

    Jensen, Helle; Andresen, Lars; Nielsen, Jens; Christensen, Jan Pravsgaard; Skov, Søren

    2011-01-01

    Vesicular stomatitis virus (VSV) has recently gained attention for its oncolytic ability in cancer treatment. Initially, we hypothesized that VSV infection could increase immune recognition of cancer cells through induction of the immune stimulatory NKG2D-ligands. Here we show that VSV infection leads to a robust induction of MICA mRNA expression, however the subsequent surface expression is potently hindered. Thus, VSV lines up with human cytomegalovirus (HCMV) and adenovirus, which actively subvert the immune system by negatively affecting NKG2D-ligand surface expression. VSV infection caused an active suppression of NKG2D-ligand surface expression, affecting both endogenous and histone deacetylase (HDAC)-inhibitor induced MICA, MICB and ULBP-2 expression. The classical immune escape mechanism of VSV (i.e., the M protein blockade of nucleocytoplasmic mRNA transport) was not involved, as the VSV mutant strain, VSV(ΔM51), which possess a defective M protein, prevented MICA surface expression similarly to wild-type VSV. The VSV mediated down modulation of NKG2D-ligand expression did not involve apoptosis. Constitutive expression of MICA bypassed the escape mechanism, suggesting that VSV affect NKG2D-ligand expression at an early post-transcriptional level. Our results show that VSV possess an escape mechanism, which could affect the immune recognition of VSV infected cancer cells. This may also have implications for immune recognition of cancer cells after combined treatment with VSV and chemotherapeutic drugs.

  8. Assessment of potential health impacts on surface water sources in Northern Nigeria

    Directory of Open Access Journals (Sweden)

    ZD Umar

    2014-05-01

    Full Text Available Northern Nigeria represents the most populous region in Nigeria and adequate water supply and sanitation typically leaves a lot to be desired in the region in order to prevent water related diseases to the vulnerable communities of the region. This research work was carried out in Kano state with specific emphasis on Tomas dam which is one of the largest dams established for multipurpose use in Nigeria. The research was aimed at evaluating Physico-chemical and Bacteriological quality of Tomas Dam in Kano State. 240 water samples from 5 sampling stations within eight months were analyzed for bacteriological and physico-chemical quality. The physico-chemical parameters indicated Biochemical Oxygen Demand (BOD and Water Current Speed results were not significantly different (P>0.05, while other physico-chemical parameters analyzed varied significantly (P<0.05. The study also revealed that Dissolved Oxygen (DO, BOD, Turbidity, and Nitrates recorded higher values than WHO and Federal Environmental Protection Agency, Nigeria (FEPA limits. Total aerobic bacterial counts (TBC and total coliform counts (TCC were determined using pour plate and Most Probable Number (MPN techniques. TBC and TCC were high and exceeded acceptable limits. Faecal coliforms, faecal Streptococciand Klebsiella species constituted 25% of the indicator organisms identified.The observed parameters indicated the Dam was contaminated with pathogenic bacteria. Variations in the weather conditions, animal and anthropogenic interferences were all directly or indirectly related to faecal contamination in the dam. Preventing indiscriminate faecal discharge close to the Dam and sufficient water treatment before consumption are of prime importance to public health as the dam was established for agricultural purposes. DOI: http://dx.doi.org/10.3126/ije.v3i2.10523 International Journal of the Environment Vol.3(2 2014: 125-136

  9. Potential Environmental Factors Affecting Oil-Degrading Bacterial Populations in Deep and Surface Waters of the Northern Gulf of Mexico.

    Science.gov (United States)

    Liu, Jiqing; Bacosa, Hernando P; Liu, Zhanfei

    2016-01-01

    Understanding bacterial community dynamics as a result of an oil spill is important for predicting the fate of oil released to the environment and developing bioremediation strategies in the Gulf of Mexico. In this study, we aimed to elucidate the roles of temperature, water chemistry (nutrients), and initial bacterial community in selecting oil degraders through a series of incubation experiments. Surface (2 m) and bottom (1537 m) waters, collected near the Deepwater Horizon site, were amended with 200 ppm light Louisiana sweet crude oil and bacterial inoculums from surface or bottom water, and incubated at 4 or 24°C for 50 days. Bacterial community and residual oil were analyzed by pyrosequencing and gas chromatography-mass spectrometry (GC-MS), respectively. The results showed that temperature played a key role in selecting oil-degrading bacteria. Incubation at 4°C favored the development of Cycloclasticus, Pseudoalteromonas , Sulfitobacter , and Reinekea , while 24°C incubations enhanced Oleibacter, Thalassobius, Phaeobacter, and Roseobacter. Water chemistry and the initial community also had potential roles in the development of hydrocarbon-degrading bacterial communities. Pseudoalteromonas , Oleibacter , and Winogradskyella developed well in the nutrient-enriched bottom water, while Reinekea and Thalassobius were favored by low-nutrient surface water. We revealed that the combination of 4°C, crude oil and bottom inoculum was a key factor for the growth of Cycloclasticus , while the combination of surface inoculum and bottom water chemistry was important for the growth of Pseudoalteromonas . Moreover, regardless of the source of inoculum, bottom water at 24°C was a favorable condition for Oleibacter. Redundancy analysis further showed that temperature and initial community explained 57 and 19% of the variation observed, while oil and water chemistry contributed 14 and 10%, respectively. Overall, this study revealed the relative roles of temperature, water

  10. Remote sensing of coastal sea-surface features off northern British Columbia

    International Nuclear Information System (INIS)

    Jardine, I.D.; Thomson, K.A.; LeBlond, P.H.; Foreman, M.G.

    1993-01-01

    This article presents an overview of surface oceanographic features identified by AVHRR imagery in Hecate Strait and adjacent waters surrounding the Queen Charlotte Islands, Canada, an area still poor in in situ observations. The observed features and their temporal variability are interpreted in terms of meteorological and hydrological forcing. The effects of tidal mixing are discussed through the application of a finite element numerical model

  11. The promoter for a variant surface glycoprotein gene expression site in Trypanosoma brucei

    NARCIS (Netherlands)

    Zomerdijk, J. C.; Ouellette, M.; ten Asbroek, A. L.; Kieft, R.; Bommer, A. M.; Clayton, C. E.; Borst, P.

    1990-01-01

    The variant-specific surface glycoprotein (VSG) gene 221 of Trypanosoma brucei is transcribed as part of a 60 kb expression site (ES). We have identified the promoter controlling this multigene transcription unit by the use of 221 chromosome-enriched DNA libraries and VSG gene 221 expression site

  12. Analysis of the precipitation and streamflow extremes in Northern Italy using high resolution reanalysis dataset Express-Hydro

    Science.gov (United States)

    Silvestro, Francesco; Parodi, Antonio; Campo, Lorenzo

    2017-04-01

    The characterization of the hydrometeorological extremes, both in terms of rainfall and streamflow, in a given region plays a key role in the environmental monitoring provided by the flood alert services. In last years meteorological simulations (both near real-time and historical reanalysis) were available at increasing spatial and temporal resolutions, making possible long-period hydrological reanalysis in which the meteo dataset is used as input in distributed hydrological models. In this work, a very high resolution meteorological reanalysis dataset, namely Express-Hydro (CIMA, ISAC-CNR, GAUSS Special Project PR45DE), was employed as input in the hydrological model Continuum in order to produce long time series of streamflows in the Liguria territory, located in the Northern part of Italy. The original dataset covers the whole Europe territory in the 1979-2008 period, at 4 km of spatial resolution and 3 hours of time resolution. Analyses in terms of comparison between the rainfall estimated by the dataset and the observations (available from the local raingauges network) were carried out, and a bias correction was also performed in order to better match the observed climatology. An extreme analysis was eventually carried on the streamflows time series obtained by the simulations, by comparing them with the results of the same hydrological model fed with the observed time series of rainfall. The results of the analysis are shown and discussed.

  13. FAP-1 Association with Fas (Apo-1) Inhibits Fas Expression on the Cell Surface

    Science.gov (United States)

    Ivanov, Vladimir N.; Bergami, Pablo Lopez; Maulit, Gabriel; Sato, Taka-Aki; Sassoon, David; Ronai, Ze'ev

    2003-01-01

    As revealed by intracellular pools of nonactive Fas (Apo-1), export of Fas to the cell surface is often impaired in human tumors, thereby inactivating Fas ligand-mediated apoptosis. Here, we demonstrate that association with Fas-associated phosphatase 1 (FAP-1) attenuates Fas export to the cell surface. Forced expression of FAP-1 reduces cell surface Fas levels and increases the intracellular pool of Fas within the cytoskeleton network. Conversely, expression of dominant-negative forms of FAP-1, or inhibition of FAP-1 expression by short interfering RNA, efficiently up-regulates surface expression of Fas. Inhibition of Fas surface expression by FAP-1 depends on its association with the C terminus of Fas. Mutation within amino acid 275 results in decreased association with FAP-1 and greater export of Fas to the cell surface in melanomas, normal fibroblasts, or Fas null cells. Identifying the role of FAP-1 in binding to, and consequently inhibition of, Fas export to the cell surface provides novel insight into the mechanism underlying the regulation of Fas trafficking, which is commonly impaired in advanced tumors with FAP-1 overexpression. PMID:12724420

  14. Systemic alteration of cell-surface and secreted glycoprotein expression in malignant breast cancer cell lines

    OpenAIRE

    Timpe, Leslie C; Yen, Roger; Haste, Nicole V; Litsakos-Cheung, Christina; Yen, Ten-Yang; Macher, Bruce A

    2013-01-01

    Breast cancer cell lines express fewer transmembrane and secreted glycoproteins than nonmalignant ones. The objective of these experiments was to characterize the changes in the expression of several hundred glycoproteins quantitatively. Secreted and cell-surface glycoproteins were isolated using a glycoprotein capture protocol and then identified by tandem mass spectrometry. Glycoproteins expressed by a group of cell lines originating from malignant tumors of the breast were compared with th...

  15. Dynamic modeling of organophosphate pesticide load in surface water in the northern San Joaquin Valley watershed of California

    International Nuclear Information System (INIS)

    Luo Yuzhou; Zhang Xuyang; Liu Xingmei; Ficklin, Darren; Zhang Minghua

    2008-01-01

    The hydrology, sediment, and pesticide transport components of the Soil and Water Assessment Tool (SWAT) were evaluated on the northern San Joaquin Valley watershed of California. The Nash-Sutcliffe coefficients for monthly stream flow and sediment load ranged from 0.49 to 0.99 over the watershed during the study period of 1992-2005. The calibrated SWAT model was applied to simulate fate and transport processes of two organophosphate pesticides of diazinon and chlorpyrifos at watershed scale. The model generated satisfactory predictions of dissolved pesticide loads relative to the monitoring data. The model also showed great success in capturing spatial patterns of dissolved diazinon and chlorpyrifos loads according to the soil properties and landscape morphology over the large agricultural watershed. This study indicated that curve number was the major factor influencing the hydrology while pesticide fate and transport were mainly affected by surface runoff and pesticide application and in the study area. - Major factors governing the instream loads of organophosphate pesticides are magnitude and timing of surface runoff and pesticide application

  16. Dynamic modeling of organophosphate pesticide load in surface water in the northern San Joaquin Valley watershed of California

    Energy Technology Data Exchange (ETDEWEB)

    Luo Yuzhou [Department of Land, Air and Water Resources, University of California, Davis, CA 95616 (United States); Institute of Watershed Science and Environmental Ecology, Wenzhou Medical College, Wenzhou, 325000 (China); Zhang Xuyang [Department of Land, Air and Water Resources, University of California, Davis, CA 95616 (United States); Liu Xingmei [Department of Land, Air and Water Resources, University of California, Davis, CA 95616 (United States); Institute of Soil, Water and Environmental Science, Zhejiang University, Hangzhou 310029 (China); Ficklin, Darren [Department of Land, Air and Water Resources, University of California, Davis, CA 95616 (United States); Zhang Minghua [Department of Land, Air and Water Resources, University of California, Davis, CA 95616 (United States); Institute of Watershed Science and Environmental Ecology, Wenzhou Medical College, Wenzhou, 325000 (China)], E-mail: mhzhang@ucdavis.edu

    2008-12-15

    The hydrology, sediment, and pesticide transport components of the Soil and Water Assessment Tool (SWAT) were evaluated on the northern San Joaquin Valley watershed of California. The Nash-Sutcliffe coefficients for monthly stream flow and sediment load ranged from 0.49 to 0.99 over the watershed during the study period of 1992-2005. The calibrated SWAT model was applied to simulate fate and transport processes of two organophosphate pesticides of diazinon and chlorpyrifos at watershed scale. The model generated satisfactory predictions of dissolved pesticide loads relative to the monitoring data. The model also showed great success in capturing spatial patterns of dissolved diazinon and chlorpyrifos loads according to the soil properties and landscape morphology over the large agricultural watershed. This study indicated that curve number was the major factor influencing the hydrology while pesticide fate and transport were mainly affected by surface runoff and pesticide application and in the study area. - Major factors governing the instream loads of organophosphate pesticides are magnitude and timing of surface runoff and pesticide application.

  17. Impacts of Irrigation on the Heat Fluxes and Near-Surface Temperature in an Inland Irrigation Area of Northern China

    Directory of Open Access Journals (Sweden)

    Li Jiang

    2014-03-01

    Full Text Available Irrigated agriculture has the potential to alter regional to global climate significantly. We investigate how irrigation will affect regional climate in the future in an inland irrigation area of northern China, focusing on its effects on heat fluxes and near-surface temperature. Using the Weather Research and Forecasting (WRF model, we compare simulations among three land cover scenarios: the control scenario (CON, the irrigation scenario (IRR, and the irrigated cropland expansion scenario (ICE. Our results show that the surface energy budgets and temperature are sensitive to changes in the extent and spatial pattern of irrigated land. Conversion to irrigated agriculture at the contemporary scale leads to an increase in annual mean latent heat fluxes of 12.10 W m−2, a decrease in annual mean sensible heat fluxes of 8.85 W m−2, and a decrease in annual mean temperature of 1.3 °C across the study region. Further expansion of irrigated land increases annual mean latent heat fluxes by 18.08 W m−2, decreases annual mean sensible heat fluxes by 12.31 W m−2, and decreases annual mean temperature by 1.7 °C. Our simulated effects of irrigation show that changes in land use management such as irrigation can be an important component of climate change and need to be considered together with greenhouse forcing in climate change assessments.

  18. Thermal infrared remote sensing in assessing groundwater and surface-water resources related to Hannukainen mining development site, northern Finland

    Science.gov (United States)

    Rautio, Anne B.; Korkka-Niemi, Kirsti I.; Salonen, Veli-Pekka

    2018-02-01

    Mining development sites occasionally host complicated aquifer systems with notable connections to natural surface water (SW) bodies. A low-altitude thermal infrared (TIR) imaging survey was conducted to identify hydraulic connections between aquifers and rivers and to map spatial surface temperature patterns along the subarctic rivers in the proximity of the Hannukainen mining development area, northern Finland. In addition to TIR data, stable isotopic compositions ( δ 18O, δD) and dissolved silica concentrations were used as tracers to verify the observed groundwater (GW) discharge into the river system. Based on the TIR survey, notable GW discharge into the main river channel and its tributaries (61 km altogether) was observed and over 500 GW discharge sites were located. On the basis of the survey, the longitudinal temperature patterns of the studied rivers were found to be highly variable. Hydrological and hydrogeological information is crucial in planning and siting essential mining operations, such as tailing areas, in order to prevent any undesirable environmental impacts. The observed notable GW discharge was taken into consideration in the planning of the Hannukainen mining development area. The results of this study support the use of TIR imagery in GW-SW interaction and environmental studies in extensive and remote areas with special concerns for water-related issues but lacking the baseline research.

  19. Sea surface cooling in the Northern South China Sea observed using Chinese Sea-wing Underwater Glider measurements

    Science.gov (United States)

    Qiu, C.; Mao, H.; Wu, J.

    2016-02-01

    Based on 26 days of Chinese Seawing underwater Glider measurements and satellite microwave data, we documented cooling of the upper mixed layer of the ocean in response to changes in the wind in the Northern South China Sea (NSCS) from September 19, 2014, to October 15, 2014. The Seawing underwater glider measured 177 profiles of temperature, salinity, and pressure within a 55 km נ55 km area, and reached a depth of 1000 m at a temporal resolution of 4 h. The study area experienced two cooling events, Cooling I and Cooling II, according to their timing. During Cooling I, water temperature at 1m depth (T1) decreased by 1.0°C, and the corresponding satellitederived surface winds increased locally by 4.2 m/s. During Cooling II, T1 decreased sharply by 1.7°C within a period of 4 days; sea surface winds increased by 7 m/s and covered the entire NSCS. The corresponding mixed layer depth (MLD) deepened sharply from 30 m to 60 m during Cooling II, and remained steady during Cooling I. We estimated temperature tendencies using a ML model. High resolution Seawing underwater glider measurements provided an estimation of MLD migration, allowing us to obtain the temporal entrainment rate of cool sub thermocline water. Quantitative analysis confirmed that the entrainment rate and latent heat flux were the two major components that regulated cooling of the ML, and that the Ekman advection and sensible heat flux were small.

  20. Mapping Surface Water DOC in the Northern Gulf of Mexico Using CDOM Absorption Coefficients and Remote Sensing Imagery

    Science.gov (United States)

    Kelly, B.; Chelsky, A.; Bulygina, E.; Roberts, B. J.

    2017-12-01

    Remote sensing techniques have become valuable tools to researchers, providing the capability to measure and visualize important parameters without the need for time or resource intensive sampling trips. Relationships between dissolved organic carbon (DOC), colored dissolved organic matter (CDOM) and spectral data have been used to remotely sense DOC concentrations in riverine systems, however, this approach has not been applied to the northern Gulf of Mexico (GoM) and needs to be tested to determine how accurate these relationships are in riverine-dominated shelf systems. In April, July, and October 2017 we sampled surface water from 80+ sites over an area of 100,000 km2 along the Louisiana-Texas shelf in the northern GoM. DOC concentrations were measured on filtered water samples using a Shimadzu TOC-VCSH analyzer using standard techniques. Additionally, DOC concentrations were estimated from CDOM absorption coefficients of filtered water samples on a UV-Vis spectrophotometer using a modification of the methods of Fichot and Benner (2011). These values were regressed against Landsat visible band spectral data for those same locations to establish a relationship between the spectral data, CDOM absorption coefficients. This allowed us to spatially map CDOM absorption coefficients in the Gulf of Mexico using the Landsat spectral data in GIS. We then used a multiple linear regressions model to derive DOC concentrations from the CDOM absorption coefficients and applied those to our map. This study provides an evaluation of the viability of scaling up CDOM absorption coefficient and remote-sensing derived estimates of DOC concentrations to the scale of the LA-TX shelf ecosystem.

  1. Heavy metals in the surface sediments of the northern portion of the South China Sea shelf: distribution, contamination, and sources.

    Science.gov (United States)

    Xu, Fangjian; Tian, Xu; Yin, Feng; Zhao, Yongfang; Yin, Xuebo

    2016-05-01

    The concentrations of seven heavy metals (Cr, Ni, Cu, Zn, As, Cd, and Pb) in the surface sediments of the northern portion of the South China Sea (SCS) shelf collected between 2012 and 2014 were measured to assess the potential contamination levels and determine the environmental risks that are associated with heavy metals in the area. The measured concentrations in the sediments were 12.4-72.5 mg kg(-1) for Cr, 4.4-29.2 mg kg(-1) for Ni, 7.1-38.1 mg kg(-1) for Cu, 19.3-92.5 mg kg(-1) for Zn, 1.3-12.1 mg kg(-1) for As, 0.03-0.24 mg kg(-1) for Cd, and 8.5-24.4 mg kg(-1) for Pb. These results indicate that the heavy metal concentrations in the sediments generally meet the China Marine Sediment Quality criteria and suggest that the overall sediment quality of the northern portion of the SCS shelf has not been significantly impacted by heavy metal pollution. However, the enrichment factor (EF) and geoaccumulation index (I geo) clearly show that elevated concentrations of Cd occur in the region. A Pearson's correlation analysis was performed, and the results suggest that Cr, Ni, Cu, and Zn have a natural origin; Cd is primarily sourced from anthropogenic activities, with partial lithogenic components, and As and Pb may be affected by factors such as varying input sources or pathways (i.e., coal burning activities and aerosol precipitation). Heavy metal contamination mostly occurred to the east of Hainan Island, mainly because of the rapid economic and social developments in the Hainan Island. The results of this study will be useful for marine environment managers for the remediation of pollution sources.

  2. Concentrations and sources of polycyclic aromatic hydrocarbons in surface coastal sediments of the northern Gulf of Mexico

    Science.gov (United States)

    2014-01-01

    Background Coastal sediments in the northern Gulf of Mexico have a high potential of being contaminated by petroleum hydrocarbons, such as polycyclic aromatic hydrocarbons (PAHs), due to extensive petroleum exploration and transportation activities. In this study we evaluated the spatial distribution and contamination sources of PAHs, as well as the bioavailable fraction in the bulk PAH pool, in surface marsh and shelf sediments (top 5 cm) of the northern Gulf of Mexico. Results PAH concentrations in this region ranged from 100 to 856 ng g−1, with the highest concentrations in Mississippi River mouth sediments followed by marsh sediments and then the lowest concentrations in shelf sediments. The PAH concentrations correlated positively with atomic C/N ratios of sedimentary organic matter (OM), suggesting that terrestrial OM preferentially sorbs PAHs relative to marine OM. PAHs with 2 rings were more abundant than those with 5–6 rings in continental shelf sediments, while the opposite was found in marsh sediments. This distribution pattern suggests different contamination sources between shelf and marsh sediments. Based on diagnostic ratios of PAH isomers and principal component analysis, shelf sediment PAHs were petrogenic and those from marsh sediments were pyrogenic. The proportions of bioavailable PAHs in total PAHs were low, ranging from 0.02% to 0.06%, with higher fractions found in marsh than shelf sediments. Conclusion PAH distribution and composition differences between marsh and shelf sediments were influenced by grain size, contamination sources, and the types of organic matter associated with PAHs. Concentrations of PAHs in the study area were below effects low-range, suggesting a low risk to organisms and limited transfer of PAHs into food web. From the source analysis, PAHs in shelf sediments mainly originated from direct petroleum contamination, while those in marsh sediments were from combustion of fossil fuels. PMID:24641695

  3. Quantitative Expression Analysis in Brassica napus by Northern Blot Analysis and Reverse Transcription-Quantitative PCR in a Complex Experimental Setting.

    Directory of Open Access Journals (Sweden)

    Annekathrin Rumlow

    Full Text Available Analysis of gene expression is one of the major ways to better understand plant reactions to changes in environmental conditions. The comparison of many different factors influencing plant growth challenges the gene expression analysis for specific gene-targeted experiments, especially with regard to the choice of suitable reference genes. The aim of this study is to compare expression results obtained by Northern blot, semi-quantitative PCR and RT-qPCR, and to identify a reliable set of reference genes for oilseed rape (Brassica napus L. suitable for comparing gene expression under complex experimental conditions. We investigated the influence of several factors such as sulfur deficiency, different time points during the day, varying light conditions, and their interaction on gene expression in oilseed rape plants. The expression of selected reference genes was indeed influenced under these conditions in different ways. Therefore, a recently developed algorithm, called GrayNorm, was applied to validate a set of reference genes for normalizing results obtained by Northern blot analysis. After careful comparison of the three methods mentioned above, Northern blot analysis seems to be a reliable and cost-effective alternative for gene expression analysis under a complex growth regime. For using this method in a quantitative way a number of references was validated revealing that for our experiment a set of three references provides an appropriate normalization. Semi-quantitative PCR was prone to many handling errors and difficult to control while RT-qPCR was very sensitive to expression fluctuations of the reference genes.

  4. Cell-surface expression of Hsp70 on hematopoietic cancer cells after inhibition of HDAC activity

    DEFF Research Database (Denmark)

    Jensen, Helle; Andresen, Lars; Hansen, Karen Aagaard

    2009-01-01

    surface-negative despite effective induction of apoptosis. Interestingly, inhibition of endolysosomes or normal ER/Golgi transport did not affect Hsp70 surface expression. Intracellular calcium and the transcription factor Sp1, which has been shown previously to be important for the intracellular stress...

  5. Reconstructing Holocene sea surface salinity changes in the Northern Aegean Sea: evidence from morphological variations of Emiliania huxleyi-coccoliths

    Science.gov (United States)

    Herrle, Jens O.; Gebühr, Christina; Bollmann, Jörg; Giesenberg, Annika; Kranzdorf, Philip

    2013-04-01

    The Aegean Sea is a key area for our understanding of the impact of changes in the hydrological cycle on ocean circulation in the Mediterranean Sea. The Aegean Sea appears to be very sensitive to climate changes in Europe because of its small volume and the position between high- and low-latitude climate regimes. Therefore, it is assumed to record environmental change, especially changes in sea surface water salinity (SSS) without a significant time lag with respect to the forcing process (Rohling et al., 2002). However, up to date, SSS cannot be easily reconstructed from geological archives because several assumptions need to be made that lead to a significant error of the salinity estimates (e.g. Rohling, 2000). Here, we present the first high resolution SSS reconstruction from a Holocene sediment core based on a recently developed transfer function using the morphological variation of Emiliania huxleyi coccoliths (Bollmann & Herrle 2007, Bollmann et al., 2009). The core is located in the northern Aegean Sea (eastern Mediterranean Basin) and covers the time period 3 -11ka ago. Sea surface water salinity in the Aegean Sea has varied in concert with temperature oscillations as recorded in Greenland ice cores (iGISP2 ice core δ18O record) with a periodicity of about 900 years (Schulz & Paull, 2002). Four major SSS events can be identified at about 3.9, 4.7, 6.4, 7.4, and 8.2 ka in the northern Aegean Sea that correlate with increases in GISP2 δ18O (Schulz & Paull, 2002) as well as decreasing percentages of tree pollen studied at the same core expect for 3.9 ka (Kotthoff et al., 2008). The most prominent salinity increase occurred during the short-lived 8.2 kyr cold event (e.g., Rohling & Pälike, 2005), which was most likely triggered by a melt-water related perturbation of the Atlantic Meridional Overturning and associated decrease of ocean heat transport to the North Atlantic. We suggest that the salinity fluctuations in the northern Aegean Sea are related to

  6. Tracking Dynamic Northern Surface Water Changes with High-Frequency Planet CubeSat Imagery

    Directory of Open Access Journals (Sweden)

    Sarah W. Cooley

    2017-12-01

    Full Text Available Recent deployments of CubeSat imagers by companies such as Planet may advance hydrological remote sensing by providing an unprecedented combination of high temporal and high spatial resolution imagery at the global scale. With approximately 170 CubeSats orbiting at full operational capacity, the Planet CubeSat constellation currently offers an average revisit time of <1 day for the Arctic and near-daily revisit time globally at 3 m spatial resolution. Such data have numerous potential applications for water resource monitoring, hydrologic modeling and hydrologic research. Here we evaluate Planet CubeSat imaging capabilities and potential scientific utility for surface water studies in the Yukon Flats, a large sub-Arctic wetland in north central Alaska. We find that surface water areas delineated from Planet imagery have a normalized root mean square error (NRMSE of <11% and geolocation accuracy of <10 m as compared with manual delineations from high resolution (0.3–0.5 m WorldView-2/3 panchromatic satellite imagery. For a 625 km2 subarea of the Yukon Flats, our time series analysis reveals that roughly one quarter of 268 lakes analyzed responded to changes in Yukon River discharge over the period 23 June–1 October 2016, one half steadily contracted, and one quarter remained unchanged. The spatial pattern of observed lake changes is heterogeneous. While connections to Yukon River control the hydrologically connected lakes, the behavior of other lakes is complex, likely driven by a combination of intricate flow paths, underlying geology and permafrost. Limitations of Planet CubeSat imagery include a lack of an automated cloud mask, geolocation inaccuracies, and inconsistent radiometric calibration across multiple platforms. Although these challenges must be addressed before Planet CubeSat imagery can achieve its full potential for large-scale hydrologic research, we conclude that CubeSat imagery offers a powerful new tool for the study and

  7. Synthetic Strategies for Semiconductor Nanocrystals Expressing Localized Surface Plasmon Resonance.

    Science.gov (United States)

    Niezgoda, J Scott; Rosenthal, Sandra J

    2016-03-03

    The field of semiconductor plasmonics has grown rapidly since its outset, only roughly six years ago, and now includes many crystalline substances ranging from GeTe to wide-bandgap transition-metal oxides. One byproduct of this proliferation is the sea of differing synthetic methods to realize localized surface plasmon resonances (LSPRs) based on the studied material. Strategies vary widely from material to material, but all have the common goal of introducing extremely high carrier densities to the semiconductor system. This doping results in tunable, size-quantized, and on/off-switchable LSPR modes, which are a complete departure from traditional metal-nanoparticle-based plasmon resonances. This Minireview will provide an overview of the current state of nanocrystal and quantum-dot plasmonics and the physical basis thereof, however its main purpose is to summarize the methods for realizing LSPRs in the various syntheses and systems that have been reported to date. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. Mapping of near surface fold structures with GPR and ERT near Steinbrunn (Northern Burgenland, Austria)

    Science.gov (United States)

    Kreutzer, Ingrid; Chwatal, Werner; Häusler, Hermann; Scheibz, Jürgen; Steirer, Fritz

    2014-05-01

    In the transition zone between the southern Vienna Basin and the Eisenstadt basin, close to Wr. Neustadt, spectacular fold structures are exposed in the former sand pit of Steinbrunn. The succession of Upper Pannonian age consists of decimetre to meter thick sandy, silty and clayey beds, which are overlain by sandstone beds (Grundtner et al., 2009). The anticline and syncline structures were interpreted as of gravitational origin by Exner et al. (2009), and reinterpreted as of tectonic origin by Häusler (2012a). In order to gain a more detailed insight to the three dimensional distribution and orientation of the folds high resolution geophysics such as electrical resistivity tomography (ERT), ground penetrating radar (GPR) and electromagnetics (EM) were applied to map the surroundings of the sandpit. The ERT- and EM-profiles show that the uppermost layer is more clayey northwest and sandier southeast of the sandpit. This is important for the GPR because clay attenuates the radar signals and therefore no clear layering of the subsurface could be mapped in these areas. In order to directly compare ERT and GPR results with the lithology of the fold structures observed in the sandpit, a reference profile on top of the 140 m long wall of the sandpit was performed. Both methods clearly reveal fold structures paralleling the folded Pannonian strata of the outcrop. While the GPR data displays boundaries and their geometry in the succession, the resistivities in the ERT portrays a more smoothened image of the observed fold structure. In almost all GPR profiles wavelike structures are visible with axes in northern direction and dome-shaped structures with axes in eastern direction, deepening towards the west. In conclusion this pattern is comparable to sections of rounded buckle folds. Although there are clayey areas wave-like and dome-like reflections can be followed in the GPR profiles over a distance of several hundred meters. This is confirmed by the ERT profiles

  9. Alzheimer's amyloid precursor protein is expressed on the surface of hematopoietic cells upon activation.

    Science.gov (United States)

    Bullido, M J; Muñoz-Fernandez, M A; Recuero, M; Fresno, M; Valdivieso, F

    1996-08-21

    A4-amyloid is the major component of senile plaques and neurofibrillary tangles found in the brain of patients suffering Alzheimer's disease. This 39-42 amino acid peptide is derived from a larger precursor protein (APP). Since APP gene encodes for a putative membrane protein, the study of APP expression at the cell surface may provide useful data for understanding its physiological function. In this report, we present data on APP expression, that was detected by APP specific mAbs in cells of the hematopoietic system. APP was weakly expressed on the cell surface of resting human lymphocytes and monocytes, but it could be induced to the surface of those cells upon stimulation. The cell activators capable of inducing APP membrane expression comprehended mitogenic lectins, calcium ionophores, phosphatase inhibitors, and anti mu-chain or anti-CD3 antibodies in B and T cells, respectively. Interestingly, phorbol esters were able to induce APP membrane expression in monocytic, but not in lymphoid cells. In contrast to lymphocytes and monocytes, granulocytes never expressed cell surface or cytoplasmic APP, even after the activation. The induction of membrane APP in response to lymphocyte activation signals, including antibodies to the antigen receptor of B and T cells, raises the possibility that APP might play the role of a cell surface receptor in the immune system.

  10. Evaluation of high-resolution satellite precipitation products with surface rain gauge observations from Laohahe Basin in northern China

    Directory of Open Access Journals (Sweden)

    Shan-hu Jiang

    2010-12-01

    Full Text Available Three high-resolution satellite precipitation products, the Tropical Rainfall Measuring Mission (TRMM standard precipitation products 3B42V6 and 3B42RT and the Climate Precipitation Center's (CPC morphing technique precipitation product (CMORPH, were evaluated against surface rain gauge observations from the Laohahe Basin in northern China. Widely used statistical validation indices and categorical statistics were adopted. The evaluations were performed at multiple time scales, ranging from daily to yearly, for the years from 2003 to 2008. The results show that all three satellite precipitation products perform very well in detecting the occurrence of precipitation events, but there are some different biases in the amount of precipitation. 3B42V6, which has a bias of 21%, fits best with the surface rain gauge observations at both daily and monthly scales, while the biases of 3B42RT and CMORPH, with values of 81% and 67%, respectively, are much higher than a normal receivable threshold. The quality of the satellite precipitation products also shows monthly and yearly variation: 3B42RT has a large positive bias in the cold season from September to April, while CMORPH has a large positive bias in the warm season from May to August, and they all attained their best values in 2006 (with 10%, 50%, and −5% biases for 3B42V6, 3B42RT, and CMORPH, respectively. Our evaluation shows that, for the Laohahe Basin, 3B42V6 has the best correspondence with the surface observations, and CMORPH performs much better than 3B42RT. The large errors of 3B42RT and CMORPH remind us of the need for new improvements to satellite precipitation retrieval algorithms or feasible bias adjusting methods.

  11. Application of near surface geophysical methods to image water table response in an Alpine Meadow, Northern California.

    Science.gov (United States)

    Ayers, M.; Blacic, T. M.; Craig, M. S.; Yarnell, S. M.

    2015-12-01

    Meadows are recognized for their value to the ecological, hydrologic, and aesthetic functions of a watershed. As natural water retention sinks, meadows attenuate floods, improve water quality and support herbaceous vegetation that stabilize streambanks and promote high biodiversity. Alpine meadows are especially vital, serving as freshwater sources and distributing to lower lying provinces through ground and surface water interaction. These complexes are highly vulnerable to drought conditions, altered seasonal precipitation patterns, and mismanaged land use. One such location, Van Norden meadow located in the Donner Summit area west of Lake Tahoe, is one of the largest sub-alpine meadows in the Sierra Nevada mountain range of Northern California. Van Norden meadow offers a natural hydrologic laboratory. Ownership transfer of the area from a local land trust to the Forestry Service requires restoration toward natural meadow conditions, and involves notching the dam in 2016 to reduce currently impounded water volumes from 250 to less than 50 acre-feet. To monitor the effects of notching the dam on the upstream meadow conditions, better understanding of the surface and groundwater hydrology both pre-and post-base level alteration is required. Comprehensive understanding of groundwater flux that supports meadow reaches relies on knowledge of their often complex stratigraphic and structural subsurface framework. In recent years hydrogeophysics has emphasized the combination of near surface geophysical techniques, collaborated with well and borehole measures, to qualitatively define these parameters. Building on a preliminary GPR investigation conducted in 2014, in which 44 270 MHz transect lines were collected, we returned to Van Norden meadow in late summer 2015 to collect lower frequency GPR (50 and 100 MHz) and electrical resistivity profiles to better define the groundwater table, sedimentary, and structural features of the meadow.

  12. Northern dwarf and low surface brightness galaxies. II - The Green Bank neutral hydrogen survey

    Science.gov (United States)

    Schneider, Stephen E.; Thuan, Trinh X.; Mangum, Jeffrey G.; Miller, John

    1992-01-01

    The paper reports neutral hydrogen observations of a large sample of dwarf and other low surface brightness galaxies. A detailed discussion and error analysis of the observations are presented, and spectra are displayed for 329 galaxies detected for the first time, or detected with substantially better signal-to-noise ratios than achieved previously. The positions on the sky of 667 galaxies meeting the present selection criteria north of delta = 38 deg are shown. The distribution of the redshifts of galaxies detected at Green Bank is illustrated. The Green Bank detections tapered off strongly below the median H I flux of 3.7 Jy km/s detected at Arecibo: only 12 percent of the Green Bank sample was detected with smaller fluxes.

  13. Measuring wintertime surface fluxes at the Tiksi observatory in northern Sakha (Yakutia)

    Science.gov (United States)

    Laurila, Thomas; Aurela, Mika; Hatakka, Juha; Tuovinen, Juha-Pekka; Asmi, Eija; Kondratyev, Vladimir; Ivakhov, Victor; Reshetnikov, Alexander; Makshtas, Alexander; Uttal, Taneil

    2013-04-01

    Tiksi hydrometeorological observatory has been equipped by new instrumentation for meteorology, turbulence, trace gas and aerosols studies as a joint effort by National Oceanic and Atmospheric Administration (NOAA), Roshydromet (Yakutian Hydrometeorological Service, Arctic and Antarctic Research Institute and Voeikov Main Geophysical Observatory units) and the Finnish Meteorological Institute (FMI). The site is close to the coast of the Laptev Sea on deep permafrost soil with low tundra vegetation and patches of arctic semidesert. Near-by terrain is gently sloping to the south. Further away they are hills in the NE- and W-directions. Turbulence (3-d wind components and sonic temperature) was measured at 10 Hz by USA-1Scientific sonic by Metek, Gmbh. Concentrations of CO2 and H2O were measured by LiCor LI7000 analyzer and CH4 concentrations by Los Gatos RMT200 analyzer. Measurement height was 2.5m. Active layer freeze up took place in extended October period. Methane and carbon dioxide emissions were observed up to early December. Emissions to the atmosphere were enhanced by turbulence created by high wind speeds. Midwinter conditions existed from the end of October to the beginning of April based on rather constant negative net radiation between 20-30 Wm-2 that cools the surface and forms highly stable stratification. Weather conditions are characterized by either low or high wind speed modes. Roughly half of the time wind speed was low, below 2 ms-1. Then, katabatic winds were common and air temperature was between -40..-30°C. High wind speeds, up to 24 ms-1, were observed during synoptic disturbances which lasted typically a few days. In this presentation we will show climatology of surface layer characteristics in late autumn and winter. We will show frequency of well-developed turbulence vs. katabatic low wind speed conditions and related atmospheric stability. The effect of wind speed on methane and carbon dioxide emissions during the freezing period will be

  14. Surface expression of NMDA receptor changes during memory consolidation in the crab Neohelice granulata

    Science.gov (United States)

    Hepp, Yanil; Salles, Angeles; Carbo-Tano, Martin

    2016-01-01

    The aim of the present study was to analyze the surface expression of the NMDA-like receptors during the consolidation of contextual learning in the crab Neohelice granulata. Memory storage is based on alterations in the strength of synaptic connections between neurons. The glutamatergic synapses undergo various forms of N-methyl-D aspartate receptor (NMDAR)-dependent changes in strength, a process that affects the abundance of other receptors at the synapse and underlies some forms of learning and memory. Here we propose a direct regulation of the NMDAR. Changes in NMDAR's functionality might be induced by the modification of the subunit's expression or cellular trafficking. This trafficking does not only include NMDAR's movement between synaptic and extra-synaptic localizations but also the cycling between intracellular compartments and the plasma membrane, a process called surface expression. Consolidation of contextual learning affects the surface expression of the receptor without affecting its general expression. The surface expression of the GluN1 subunit of the NMDAR is down-regulated immediately after training, up-regulated 3 h after training and returns to naïve and control levels 24 h after training. The changes in NMDAR surface expression observed in the central brain are not seen in the thoracic ganglion. A similar increment in surface expression of GluN1 in the central brain is observed 3 h after administration of the competitive GABAA receptor antagonist, bicuculline. These consolidation changes are part of a plasticity event that first, during the down-regulation, stabilizes the trace and later, at 3-h post-training, changes the threshold for synapse activation. PMID:27421895

  15. Long-term evolution of the surface environment of the Campine area, Northern Belgium

    International Nuclear Information System (INIS)

    Beerten, K.; De Craen, M.; Brassinnes, S.; Wouters, L.

    2012-01-01

    Document available in extended abstract form only. The Boom Clay in the Campine Basin is studied as the reference host formation for the geological disposal of radioactive waste. Near Mol, it is covered by approximately 200 m of sediment which protects the clay layer. Here, we will give a preliminary assessment of the long-term evolution of the surface environment in the Campine area for the next 1 Ma, with respect to geomorphological, pedological and hydrological processes. The current surface environment started to develop after the last major marine regression around 2 Ma ago. Then, the climatic and tectonic evolution caused significant changes in surficial geology, relief intensity, soils and hydrography. The palaeo-geographical evolution during the Quaternary is characterized by important stream divergences, relative uplift and erosion. Tectonic uplift of the Campine Plateau forced the river Meuse to incise into the substrate. To the west, erosional processes shaped the Nete and the Lower Scheldt basins as a result of combined uplift and gradual sea-level drop. The Campine Plateau being covered by coarse fluvial deposits from the Early and Middle Pleistocene, is largely protected from erosion. The same is true for the Campine Cuesta, that stands out as a result of the cohesive and erosion resistant Kempen Clay substrate. Burial graphs covering the last few million years from different locations in the Boom Clay sub-crop zone reflect processes of subsidence, uplift, sea level variations and climate change in relation to the erodibility of the substrate. Continuous burial during the Quaternary as a result of subsidence is observed near the Dutch border, and for the Roer Valley Graben. Burial graphs for the Campine Plateau do not show important erosional phases during the last several million years. This is due to the protecting cover of coarse fluvial deposits. The area that is now occupied by the Lower Scheldt and the Nete basin has experienced erosion during

  16. Surface ozone seasonality under global change: Influence from dry deposition and isoprene emissions at northern mid-latitudes

    Science.gov (United States)

    Clifton, O.; Paulot, F.; Fiore, A. M.; Horowitz, L. W.; Malyshev, S.; Shevliakova, E.; Correa, G. J. P.; Lin, M.

    2017-12-01

    Identifying the contributions of nonlinear chemistry and transport to observed surface ozone seasonal cycles over land using global models relies on an accurate representation of ozone uptake by vegetation (dry deposition). It is well established that in the absence of ozone precursor emission changes, a warming climate will increase surface ozone in polluted regions, and that a rise in temperature-dependent isoprene emissions would exacerbate this "climate penalty". However, the influence of changes in ozone dry deposition, expected to evolve with climate and land use, is often overlooked in air quality projections. With a new scheme that represents dry deposition within the NOAA GFDL dynamic vegetation land model (LM3) coupled to the NOAA GFDL atmospheric chemistry-climate model (AM3), we simulate the impact of 21st century climate and land use on ozone dry deposition and isoprene emissions. This dry deposition parameterization is a version of the Wesely scheme, but uses parameters explicitly calculated by LM3 that respond to climate and land use (e.g., stomatal conductance, canopy interception of water, leaf area index). The parameterization includes a nonstomatal deposition dependence on humidity. We evaluate climatological present-day seasonal cycles of ozone deposition velocities and abundances with those observed at northern mid-latitude sites. With a set of 2010s and 2090s decadal simulations under a high climate warming scenario (RCP8.5) and a sensitivity simulation with well-mixed greenhouse gases following RCP8.5 but air pollutants held at 2010 levels (RCP8.5_WMGG), we examine changes in surface ozone seasonal cycles. We build on our previous findings, which indicate that strong reductions in anthropogenic NOx emissions under RCP8.5 cause the surface ozone seasonal cycle over the NE USA to reverse, shifting from a summer peak at present to a winter peak by 2100. Under RCP8.5_WMGG, we parse the separate effects of climate and land use on ozone dry

  17. Pool patterning in a northern peatland using near surface geophysics: the role of glacial deposition

    Science.gov (United States)

    Comas, X.; Slater, L.; Reeve, A.; Nolan, J.; Jol, H.

    2008-05-01

    The potential role of stratigraphy and lithology on the processes leading to pool formation were examined in Caribou Bog, a 2200-hectare peatland in central Maine. The area is surrounded by esker deposit outcrops from the Katahdin system, that extends 150 km from central Maine to the coast (orientated approximately N-S), and is dominated by sharp-crested eskers with poorly sorted sand, gravel and boulders. A combination of hydrogeophysical techniques were used to examine the correlation between pool location within the bog and subsurface stratigraphy and lithology that included: ground penetrating radar (GPR), electrical resistivity, EM-31, hydrological measurements and direct sampling. Previous studies in the area showed certain correspondence between elevated mineral soil surfaces (interpreted as buried eskers) and pool location. The work presented here expands upon our previous results by including a wider array of measurements to better constrain that correspondence. Hydrological measurements showed spatial correlation between stronger downward hydraulic gradients and proximity to the esker crests. Geophysical data along the pool area consistently showed two buried esker crests with dipping and undulating bedding, collapse structures and presence of boulders in sediment. A conceptual model for pool development that accounts for the initial (e.g. heterogeneous peat growth due to local enhanced decomposition) and final stages (e.g. lateral spreading) of pool development is proposed based on these findings.

  18. Geologic mapping of near-surface sediments in the northern Mississippi Embayment, McCracken County, KY

    Energy Technology Data Exchange (ETDEWEB)

    Sexton, Joshua L [JL Sexton and Son; Fryar, Alan E [Dept of Earth and Geoligical Sciences, Univ of KY,; Greb, s F [Univ of KY, KY Geological Survey

    2006-04-01

    POSTER: The Jackson Purchase region of western Kentucky consists of Coastal Plain sediments near the northern margin of the Mississippi Embayment. Within this region is the Paducah Gaseous Diffusion Plant (PGDP), a uranium enrichment facility operated by the US Department of Energy. At PGDP, a Superfund site, soil and groundwater studies have provided subsurface lithologic data from hundreds of monitoring wells and borings. Despite preliminary efforts by various contractors, these data have not been utilized to develop detailed stratigraphic correlations of sedimentary units across the study area. In addition, sedimentary exposures along streams in the vicinityof PGDP have not been systematically described beyond the relatively simple geologic quadrangle maps published by the US Geological Survey in 1966-67. This study integrates lithologic logs, other previous site investigation data, and outcrop mapping to provide a compilation of near-surface lithologic and stratigraphic data for the PGDP area. A database of borehole data compiled during this study has been provided to PGDP for future research and archival.

  19. Whole genome expression profiling using DNA microarray for determining biocompatibility of polymeric surfaces

    DEFF Research Database (Denmark)

    Stangegaard, Michael; Wang, Zhenyu; Kutter, Jörg Peter

    2006-01-01

    conventional methods to determine biocompatibility such as cellular growth rate, morphology and the hydrophobicity of the surfaces. HeLa cells grown on polymethylmethacrylate (PMMA) or a SU-8 surface treated with HNO3-ceric ammonium nitrate (HNO3-CAN) and ethanolamine showed no differences in growth rate......, morphology or gene expression profiles as compared to HeLa cells grown in cell culture flasks. Cells grown on SU-8 treated with only HNO3-CAN showed almost the same growth rate (36 ¡ 1 h) and similar morphology as cells grown in cell culture flasks (32 ¡ 1 h), indicating good biocompatibility. However, more...... than 200 genes showed different expression levels in cells grown on SU-8 treated with HNO3-CAN compared to cells grown in cell culture flasks. This shows that gene expression profiling is a simple and precise method for determining differences in cells grown on different surfaces that are otherwise...

  20. Estimating surface pCO2 in the northern Gulf of Mexico: Which remote sensing model to use?

    Science.gov (United States)

    Chen, Shuangling; Hu, Chuanmin; Cai, Wei-Jun; Yang, Bo

    2017-12-01

    Various approaches and models have been proposed to remotely estimate surface pCO2 in the ocean, with variable performance as they were designed for different environments. Among these, a recently developed mechanistic semi-analytical approach (MeSAA) has shown its advantage for its explicit inclusion of physical and biological forcing in the model, yet its general applicability is unknown. Here, with extensive in situ measurements of surface pCO2, the MeSAA, originally developed for the summertime East China Sea, was tested in the northern Gulf of Mexico (GOM) where river plumes dominate water's biogeochemical properties during summer. Specifically, the MeSAA-predicted surface pCO2 was estimated by combining the dominating effects of thermodynamics, river-ocean mixing and biological activities on surface pCO2. Firstly, effects of thermodynamics and river-ocean mixing (pCO2@Hmixing) were estimated with a two-endmember mixing model, assuming conservative mixing. Secondly, pCO2 variations caused by biological activities (ΔpCO2@bio) was determined through an empirical relationship between sea surface temperature (SST)-normalized pCO2 and MODIS (Moderate Resolution Imaging Spectroradiometer) 8-day composite chlorophyll concentration (CHL). The MeSAA-modeled pCO2 (sum of pCO2@Hmixing and ΔpCO2@bio) was compared with the field-measured pCO2. The Root Mean Square Error (RMSE) was 22.94 μatm (5.91%), with coefficient of determination (R2) of 0.25, mean bias (MB) of - 0.23 μatm and mean ratio (MR) of 1.001, for pCO2 ranging between 316 and 452 μatm. To improve the model performance, a locally tuned MeSAA was developed through the use of a locally tuned ΔpCO2@bio term. A multi-variate empirical regression model was also developed using the same dataset. Both the locally tuned MeSAA and the regression models showed improved performance comparing to the original MeSAA, with R2 of 0.78 and 0.84, RMSE of 12.36 μatm (3.14%) and 10.66 μatm (2.68%), MB of 0.00 μatm and - 0

  1. Land surface temperature changes in Northern Iberia since 4000 yr BP, based on δ13C of speleothems

    Science.gov (United States)

    Martín-Chivelet, Javier; Muñoz-García, M. Belén; Edwards, R. Lawrence; Turrero, María J.; Ortega, Ana I.

    2011-05-01

    The surface temperature changes for the last 4000 years in northern inland Iberia (an area particularly sensitive to climate change) are determined by a high resolution study of carbon stable isotope records of stalagmites from three caves (Kaite, Cueva del Cobre, and Cueva Mayor) separated several tens of kilometers away in N Spain. Despite the local conditions of each cave, the isotopic series show a good overall coherence, and resulted to be strongly sensitive to surface temperature changes. The record reflects alternating warmer and colder intervals, always within a temperature range of 1.6 °C. The timing and duration of the intervals were provided by 43 230Th- 234U (ICP-MS) ages. Main climatic recognized periods are: (1) 3950-3000 yr BP: warm period punctuated by cool events around ~ 3950, 3550 and 3250 yr BP; (2) 2850-2500 yr BP cold interval (Iron Age Cold Period); (3) 2500-1650 yr BP moderate warm period (Roman Warm Period), with maximum temperatures between 2150 and 1750 yr BP; (4) 1650-1350 yr BP cold interval (Dark Ages Cold Period), with a thermal minimum at ~ 1500 yr BP; (5) 1350-750 yr BP warm period (Medieval Warm Period) punctuated by two cooler events at ~ 1250 and ~ 850 yr BP; (6) 750-100 yr BP cold period (Little Ice Age) with extremes occurring at 600-500 yr BP, 350-300 yr BP, and 150-100 yr BP; and (7) the last 150 years, characterized by rapid but no linear warming (Modern Warming). Remarkably, the presented records allow direct comparison of recent warming with former warm intervals such as the Roman or the Medieval periods. That comparison reveals the 20th century as the time with highest surface temperatures of the last 4000 years for the studied area. Spectral analysis of the time series shows consistent climatic cycles of ~ 400, ~ 900 and ~ 1300 yr, comparable with those recognized in the North Atlantic marine record, the Greenland ice cores, and other terrestrial records for the middle-late Holocene, suggesting common climate forcing

  2. AMP-activated protein kinase downregulates Kv7.1 cell surface expression

    DEFF Research Database (Denmark)

    Andersen, Martin N; Krzystanek, Katarzyna; Jespersen, Thomas

    2012-01-01

    The potassium channel Kv7.1 is expressed in the heart, where it contributes to the repolarization of the cardiac action potential. Additionally, Kv7.1 is expressed in epithelial tissues playing a role in salt and water transport. We recently demonstrated that surface-expressed Kv7.1 is internalized...... in response to polarization of the epithelial Madin-Darby canine kidney (MDCK) cell line and that this was mediated by activation of protein kinase C (PKC). In this study, the pathway downstream of PKC, which leads to internalization of Kv7.1 upon cell polarization, is elucidated. We show by confocal...

  3. Uropathogenic Escherichia coli Express Type 1 Fimbriae Only in Surface Adherent Populations Under Physiological Growth Conditions.

    Science.gov (United States)

    Stærk, Kristian; Khandige, Surabhi; Kolmos, Hans Jørn; Møller-Jensen, Jakob; Andersen, Thomas Emil

    2016-02-01

    Most uropathogenic Escherichia coli (UPEC) strains harbor genes encoding adhesive type 1 fimbria (T1F). T1F is a key factor for successful establishment of urinary tract infection. However, UPEC strains typically do not express T1F in the bladder urine, and little is understood about its induction in vivo. A flow chamber infection model was used to grow UPEC under conditions simulating distinct infection niches in the bladder. Type 1 fimbriation on isolated UPEC was subsequently determined by yeast cell agglutination and immunofluorescence microscopy, and the results were correlated with the ability to adhere to and invade cultured human bladder cells. Although inactive during planktonic growth in urine, T1F expression occurs when UPEC settles on and infects bladder epithelial cells or colonizes catheters. As a result, UPEC in these sessile populations enhances bladder cell adhesion and invasion potential. Only T1F-negative UPEC are subsequently released to the urine, thus limiting T1F expression to surface-associated UPEC alone. Our results demonstrate that T1F expression is strictly regulated under physiological growth conditions with increased expression during surface growth adaptation and infection of uroepithelial cells. This leads to separation of UPEC into low-expression planktonic populations and high-expression sessile populations. © The Author 2015. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail journals.permissions@oup.com.

  4. A dual tag system for facilitated detection of surface expressed proteins in Escherichia coli

    Directory of Open Access Journals (Sweden)

    Jarmander Johan

    2012-09-01

    Full Text Available Abstract Background The discovery of the autotransporter family has provided a mechanism for surface expression of proteins in laboratory strains of Escherichia coli. We have previously reported the use of the AIDA-I autotransport system to express the Salmonella enterica serovar Enteritidis proteins SefA and H:gm. The SefA protein was successfully exposed to the medium, but the orientation of H:gm in the outer membrane could not be determined due to proteolytic cleavage of the N-terminal detection-tag. The goal of the present work was therefore to construct a vector containing elements that facilitates analysis of surface expression, especially for proteins that are sensitive to proteolysis or otherwise difficult to express. Results The surface expression system pAIDA1 was created with two detection tags flanking the passenger protein. Successful expression of SefA and H:gm on the surface of E. coli was confirmed with fluorescently labeled antibodies specific for the N-terminal His6-tag and the C-terminal Myc-tag. While both tags were detected during SefA expression, only the Myc-tag could be detected for H:gm. The negative signal indicates a proteolytic cleavage of this protein that removes the His6-tag facing the medium. Conclusions Expression levels from pAIDA1 were comparable to or higher than those achieved with the formerly used vector. The presence of the Myc- but not of the His6-tag on the cell surface during H:gm expression allowed us to confirm the hypothesis that this fusion protein was present on the surface and oriented towards the cell exterior. Western blot analysis revealed degradation products of the same molecular weight for SefA and H:gm. The size of these fragments suggests that both fusion proteins have been cleaved at a specific site close to the C-terminal end of the passenger. This proteolysis was concluded to take place either in the outer membrane or in the periplasm. Since H:gm was cleaved to a much greater extent

  5. Linking crop structure, throughfall, soil surface conditions, runoff and soil detachment: 10 land uses analyzed in Northern Laos.

    Science.gov (United States)

    Lacombe, Guillaume; Valentin, Christian; Sounyafong, Phabvilay; de Rouw, Anneke; Soulileuth, Bounsamai; Silvera, Norbert; Pierret, Alain; Sengtaheuanghoung, Oloth; Ribolzi, Olivier

    2018-03-01

    In Montane Southeast Asia, deforestation and unsuitable combinations of crops and agricultural practices degrade soils at an unprecedented rate. Typically, smallholder farmers gain income from "available" land by replacing fallow or secondary forest by perennial crops. We aimed to understand how these practices increase or reduce soil erosion. Ten land uses were monitored in Northern Laos during the 2015 monsoon, using local farmers' fields. Experiments included plots of the conventional system (food crops and fallow), and land uses corresponding to new market opportunities (e.g. commercial tree plantations). Land uses were characterized by measuring plant cover and plant mean height per vegetation layer. Recorded meteorological variables included rainfall intensity, throughfall amount, throughfall kinetic energy (TKE), and raindrop size. Runoff coefficient, soil loss, and the percentage areas of soil surface types (free aggregates and gravel; crusts; macro-faunal, vegetal and pedestal features; plant litter) were derived from observations and measurements in 1-m 2 micro-plots. Relationships between these variables were explored with multiple regression analyses. Our results indicate that TKE induces soil crusting and soil loss. By reducing rainfall infiltration, crusted area enhances runoff, which removes and transports soil particles detached by splash over non-crusted areas. TKE is lower under land uses reducing the velocity of raindrops and/or preventing an increase in their size. Optimal vegetation structures combine minimum height of the lowest layer (to reduce drop velocity at ground level) and maximum coverage (to intercept the largest amount of rainfall), as exemplified by broom grass (Thysanolaena latifolia). In contrast, high canopies with large leaves will increase TKE by enlarging raindrops, as exemplified by teak trees (Tectona grandis), unless a protective understorey exists under the trees. Policies that ban the burning of multi-layered vegetation

  6. SRC Inhibition Reduces NR2B Surface Expression and Synaptic Plasticity in the Amygdala

    Science.gov (United States)

    Sinai, Laleh; Duffy, Steven; Roder, John C.

    2010-01-01

    The Src protein tyrosine kinase plays a central role in the regulation of N-methyl-d-aspartate receptor (NMDAR) activity by regulating NMDAR subunit 2B (NR2B) surface expression. In the amygdala, NMDA-dependent synaptic plasticity resulting from convergent somatosensory and auditory inputs contributes to emotional memory; however, the role of Src…

  7. Processing OMEGA/Mars Express hyperspectral imagery from radiance-at-sensor to surface reflectance

    NARCIS (Netherlands)

    Bakker, W.H.; Ruitenbeek, F.J.A. van; Werff, H.M.A. van der; Zegers, T.E.; Oosthoek, J.H.P.; Marsh, S.H.; Meer, F.D. van der

    2014-01-01

    OMEGA/Mars Express hyperspectral imagery is an excellent source of data for exploring the surface composition of the planet Mars. Compared to terrestrial hyperspectral imagery, the data are challenging to work with; scene-specific transmission models are lacking, spectral features are shallow making

  8. Basement to surface expressions and critical factors in the genesis of unconformity-related deposits

    International Nuclear Information System (INIS)

    Potter, Eric

    2014-01-01

    Two subprojects: 1) Basement to surface expressions of deep mineralization and refinement of critical factors leading to the genesis of unconformity-related uranium deposits; and 2) Recognition of uranium ore system alteration signatures in complex terranes: IOCG vs albite-hosted uranium vs volcanic-hosted uranium.

  9. Surface Expression of NMDA Receptor Changes during Memory Consolidation in the Crab "Neohelice granulata"

    Science.gov (United States)

    Hepp, Yanil; Salles, Angeles; Carbo-Tano, Martin; Pedreira, Maria Eugenia; Freudenthal, Ramiro

    2016-01-01

    The aim of the present study was to analyze the surface expression of the NMDA-like receptors during the consolidation of contextual learning in the crab "Neohelice granulata". Memory storage is based on alterations in the strength of synaptic connections between neurons. The glutamatergic synapses undergo various forms of…

  10. The Effect of Laminin-1-Doped Nanoroughened Implant Surfaces: Gene Expression and Morphological Evaluation

    Directory of Open Access Journals (Sweden)

    Humberto Osvaldo Schwartz-Filho

    2012-01-01

    Full Text Available Aim. This study aimed to observe the morphological and molecular effect of laminin-1 doping to nanostructured implant surfaces in a rabbit model. Materials and Methods. Nanostructured implants were coated with laminin-1 (test; dilution, 100 μg/mL and inserted into the rabbit tibiae. Noncoated implants were used as controls. After 2 weeks of healing, the implants were removed and subjected to morphological analysis using scanning electron microscopy (SEM and gene expression analysis using the real-time reverse transcriptase-polymerase chain reaction (RT-PCR. Results. SEM revealed bony tissue attachment for both control and test implants. Real-time RT-PCR analysis showed that the expression of osteoblast markers RUNX-2, osteocalcin, alkaline phosphatase, and collagen I was higher (1.62-fold, 1.53-fold, 1.97-fold, and 1.04-fold, resp. for the implants modified by laminin-1 relative to the control. All osteoclast markers investigated in the study presented higher expression on the test implants than controls as follows: tartrate-resistant acid phosphatase (1.67-fold, calcitonin receptor (1.35-fold, and ATPase (1.25-fold. The test implants demonstrated higher expression of inflammatory markers interleukin-10 (1.53-fold and tumour necrosis factor-α (1.61-fold relative to controls. Conclusion. The protein-doped surface showed higher gene expression of typical genes involved in the osseointegration cascade than the control surface.

  11. Dissolved methane concentrations in the water column and surface sediments of Hanna Shoal and Barrow Canyon, Northern Chukchi Sea

    Science.gov (United States)

    Lapham, Laura; Marshall, Kathleen; Magen, Cédric; Lyubchich, Viacheslav; Cooper, Lee W.; Grebmeier, Jacqueline M.

    2017-10-01

    Current estimates of methane (CH4) flux suggest that Arctic shelves may be a significant source of atmospheric CH4, a potent greenhouse gas. However, little information is known about the CH4 flux from most Arctic shelves, other than the East Siberian Arctic Shelf. We report here dissolved CH4 concentrations in the water column and within surface sediments of the Northern Chukchi Sea. We hypothesized that this area contains high concentrations of CH4 because it receives nutrient rich waters through the Bering Strait, promoting primary production that enhances an organic-rich material flux to the seafloor and eventual microbial methanogenesis in the sediments. In August 2012, as part of the Chukchi Sea Offshore Monitoring in Drilling Area (COMIDA) project, fourteen stations were sampled on Hanna Shoal, a shallow feature on the shelf, and ten stations across the undersea Barrow Canyon. On Hanna Shoal, water column CH4 concentrations ranged from 14 to 74 nM, and surface concentrations were up to 15 times supersaturated in CH4 compared to equilibrium with the average atmospheric concentrations (3 nM). CH4 concentrations at the sediment-water interface were around 1,500 nM, and typically increased with depth in the sediment. At the head of Barrow Canyon, water column CH4 concentrations ranged from 5 to 46 nM, with the highest concentrations in the deepest waters that were sampled (118 m). Overall, the calculated fluxes to the atmosphere ranged from 1 to 80 μmol CH4 m-2 d-1 for Hanna Shoal and 4 to 17 μmol CH4 m-2 d-1 across the Barrow Canyon stations. Although there was a large range in these fluxes, the average atmospheric flux (20 μmol CH4 m-2 d-1) across Hanna Shoal was 12 times lower than the flux reported from the East Siberian Arctic Shelf in summer. We conclude that while there is a positive flux of CH4 to the atmosphere, this part of the Chukchi Sea is not a significant source of atmospheric CH4 compared to the East Siberian Sea shelf.

  12. Expression and surface display of Cellulomonas endoglucanase in the ethanologenic bacterium Zymobacter palmae.

    Science.gov (United States)

    Kojima, Motoki; Akahoshi, Tomohiro; Okamoto, Kenji; Yanase, Hideshi

    2012-11-01

    In order to reduce the cost of bioethanol production from lignocellulosic biomass, we developed a tool for cell surface display of cellulolytic enzymes on the ethanologenic bacterium Zymobacter palmae. Z. palmae is a novel ethanol-fermenting bacterium capable of utilizing a broad range of sugar substrates, but not cellulose. Therefore, to express and display heterologous cellulolytic enzymes on the Z. palmae cell surface, we utilized the cell-surface display motif of the Pseudomonas ice nucleation protein Ina. The gene encoding Ina from Pseudomonas syringae IFO3310 was cloned, and its product was comprised of three functional domains: an N-terminal domain, a central domain with repeated amino acid residues, and a C-terminal domain. The N-terminal domain of Ina was shown to function as the anchoring motif for a green fluorescence protein fusion protein in Escherichia coli. To express a heterologous cellulolytic enzyme extracellularly in Z. palmae, we fused the N-terminal coding sequence of Ina to the coding sequence of an N-terminal-truncated Cellulomonas endoglucanase. Z. palmae cells carrying the fusion endoglucanase gene were shown to degrade carboxymethyl cellulose. Although a portion of the expressed fusion endoglucanase was released from Z. palmae cells into the culture broth, we confirmed the display of the protein on the cell surface by immunofluorescence microscopy. The results indicate that the N-terminal anchoring motif of Ina from P. syringae enabled the translocation and display of the heterologous cellulase on the cell surface of Z. palmae.

  13. Expression and surface display of Cellulomonas endoglucanase in the ethanologenic bacterium Zymobacter palmae

    Energy Technology Data Exchange (ETDEWEB)

    Kojima, Motoki; Akahoshi, Tomohiro; Okamoto, Kenji; Yanase, Hideshi [Tottori Univ. (Japan). Dept. of Chemistry and Biotechnology

    2012-11-15

    In order to reduce the cost of bioethanol production from lignocellulosic biomass, we developed a tool for cell surface display of cellulolytic enzymes on the ethanologenic bacterium Zymobacter palmae. Z. palmae is a novel ethanol-fermenting bacterium capable of utilizing a broad range of sugar substrates, but not cellulose. Therefore, to express and display heterologous cellulolytic enzymes on the Z. palmae cell surface, we utilized the cell-surface display motif of the Pseudomonas ice nucleation protein Ina. The gene encoding Ina from Pseudomonas syringae IFO3310 was cloned, and its product was comprised of three functional domains: an N-terminal domain, a central domain with repeated amino acid residues, and a C-terminal domain. The N-terminal domain of Ina was shown to function as the anchoring motif for a green fluorescence protein fusion protein in Escherichia coli. To express a heterologous cellulolytic enzyme extracellularly in Z. palmae, we fused the N-terminal coding sequence of Ina to the coding sequence of an N-terminal-truncated Cellulomonas endoglucanase. Z. palmae cells carrying the fusion endoglucanase gene were shown to degrade carboxymethyl cellulose. Although a portion of the expressed fusion endoglucanase was released from Z. palmae cells into the culture broth, we confirmed the display of the protein on the cell surface by immunofluorescence microscopy. The results indicate that the N-terminal anchoring motif of Ina from P. syringae enabled the translocation and display of the heterologous cellulase on the cell surface of Z. palmae. (orig.)

  14. Comparing the effects of atrazine and an environmentally relevant mixture on estrogen-responsive gene expression in the northern leopard frog and the fathead minnow.

    Science.gov (United States)

    Ali, Jonathan M; Knight, Lindsey A; D'Souza, Del L; Kolok, Alan S

    2018-04-01

    In Nebraska, fish are exposed to herbicides in agricultural runoff. The study objectives were to determine 1) if fathead minnows and northern leopard frogs exposed to atrazine experience alterations in gene expression, and 2) whether these changes are elicited by a simulated herbicide mixture. Following a 7-d exposure to atrazine, female minnows were defeminized, whereas male frogs were feminized. The mixture did not elicit statistically significant effects in either species. Environ Toxicol Chem 2018;37:1182-1188. © 2018 SETAC. © 2018 SETAC.

  15. Cell surface expression and function of the macromolecular C1 complex on the surface of human monocytes

    Directory of Open Access Journals (Sweden)

    Kinga K Hosszu

    2012-03-01

    Full Text Available The synthesis of the subunits of the C1 complex (C1q, C1s, C1r, and its regulator C1 inhibitor (C1-Inh by human monocytes has been previously established. However, surface expression of these molecules by monocytes has not been shown. Using flow cytometry and antigen-capture ELISA, we show here for the first time that, in addition to C1q, PB monocytes and the monocyte-derived U937 cells express C1s and C1r, as well as Factor B and C1-Inh on their surface. C1s and C1r immunoprecipitated with C1q, suggesting that at least some of the C1q on these cells is part of the C1 complex. Furthermore, the C1 complex on U937 cells was able to trigger complement activation via the classical pathway. The presence of C1-Inh may ensure that an unwarranted autoactivation of the C1 complex does not take place. Since C1-Inh closely monitors the activation of the C1 complex in a sterile or infectious inflammatory environment, further elucidation of the role of C1 complex is crucial to dissect its function in monocyte, DC and T cell activities, and its implications in host defense and tolerance.

  16. Natural radionuclides tracing in marine surface waters along the northern coast of Oman Sea by combining the radioactivity analysis, oceanic currents and the SWAN model results

    International Nuclear Information System (INIS)

    Zare, Mohammad Reza; Mostajaboddavati, Mojtaba; Kamali, Mahdi; Tari, Marziyeh; Mosayebi, Sanaz; Mortazavi, Mohammad Seddigh

    2015-01-01

    Highlights: • This study estimates radioactive pollution diffusion in coastline of the Oman Sea. • 36 high volume surface water samples were analyzed using a portable HPGe detector. • Oceanic currents in the northern coast of Oman Sea were investigated. • The spectral wave model SWAN was used for wave parameters simulation. • Currents and preferable wave directions were coupled with higher radioactivity. - Abstract: This study aims to establish a managed sampling plan for rapid estimate of natural radio-nuclides diffusion in the northern coast of the Oman Sea. First, the natural radioactivity analysis in 36 high volume surface water samples was carried out using a portable high-resolution gamma-ray spectrometry. Second, the oceanic currents in the northern coast were investigated. Then, the third generation spectral SWAN model was utilized to simulate wave parameters. Direction of natural radioactivity propagation was coupled with the preferable wave vectors and oceanic currents direction that face to any marine pollution, these last two factors will contribute to increase or decrease of pollution in each grid. The results were indicated that the natural radioactivity concentration between the grids 8600 and 8604 is gathered in the grid 8600 and between the grids 8605 and 8608 is propagated toward middle part of Oman Sea

  17. Regulation of ADAM12 cell-surface expression by protein kinase C epsilon

    DEFF Research Database (Denmark)

    Sundberg, Christina; Thodeti, Charles Kumar; Kveiborg, Marie

    2004-01-01

    The ADAM (a disintegrin and metalloprotease) family consists of multidomain cell-surface proteins that have a major impact on cell behavior. These transmembrane-anchored proteins are synthesized as proforms that have (from the N terminus): a prodomain; a metalloprotease-, disintegrin......-like-, cysteine-rich, epidermal growth factor-like, and transmembrane domain; and a cytoplasmic tail. The 90-kDa mature form of human ADAM12 is generated in the trans-Golgi through cleavage of the prodomain by a furin-peptidase and is stored intracellularly until translocation to the cell surface...... as a constitutively active protein. However, little is known about the regulation of ADAM12 cell-surface translocation. Here, we used human RD rhabdomyosarcoma cells, which express ADAM12 at the cell surface, in a temporal pattern. We report that protein kinase C (PKC) epsilon induces ADAM12 translocation to the cell...

  18. 20 January 2014 - Members of the Regional Assemblies and Parliaments United Kingdom of Great Britain and Northern Ireland visiting the LHC tunnel at Point 8 with Technology Department, Vacuum, Surfaces and Coatings Group P. Cruikshank.

    CERN Document Server

    Pantelia, Anna

    2014-01-01

    20 January 2014 - Members of the Regional Assemblies and Parliaments United Kingdom of Great Britain and Northern Ireland visiting the LHC tunnel at Point 8 with Technology Department, Vacuum, Surfaces and Coatings Group P. Cruikshank.

  19. Hydrogeochemistry and stable isotopes of ground and surface waters from two adjacent closed basins, Atacama Desert, northern Chile

    Science.gov (United States)

    Alpers, Charles N.; Whittemore, D.O.

    1990-01-01

    The geochemistry and stable isotopes of groundwaters, surface waters, and precipitation indicate different sources of some dissolved constituents, but a common source of recharge and other constituents in two adjacent closed basins in the Atacama Desert region of northern Chile (24??15???-24??45???S). Waters from artesian wells, trenches, and ephemeral streams in the Punta Negra Basin are characterized by concentrations of Na>Ca>Mg and Cl ???SO4, with TDS Mg ??? Ca and SO4 > Cl, with TDS also Mg ??? Ca and SO4 > Cl, but with TDS up to 40 g/l. The deep mine waters have pH between 3.2 and 3.9, and are high in dissolved CO2 (??13 C = -4.8%PDB), indicating probable interaction with oxidizing sulfides. The deep mine waters have ??18O values of ???-1.8%.compared with values < -3.5??? for other Hamburgo Basin waters; thus the mine waters may represent a mixture of meteoric waters with deeper "metamorphic" waters, which had interacted with rocks and exchanged oxygen isotopes at elevated temperatures. Alternatively, the deep mine waters may represent fossil meteoric waters which evolved isotopically along an evaporative trend starting from values quite depleted in ??18O and ??Dd relative to either precipitation or shallow groundwaters. High I/Br ratios in the Hamburgo Basin waters and La Escondida mine waters are consistent with regionally high I in surficial deposits in the Atacama Desert region and may represent dissolution of a wind-blown evaporite component. Rain and snow collected during June 1984, indicate systematic ??18O and ??D fractionation with increasing elevation between 3150 and 4180 m a.s.l. (-0.21??.??18O and -1.7??.??D per 100 m). Excluding the deep mine waters from La Escondida, the waters from the Hamburgo and Punta Negra Basins have similar ??D and ??18O values and together show a distinct evaporative trend (??D = 5.0 ??18O - 20.2). Snowmelt from the central Andes Cordillera to the east is the most likely source of recharge to both basins. Some of the

  20. 3D facial expression recognition based on histograms of surface differential quantities

    KAUST Repository

    Li, Huibin

    2011-01-01

    3D face models accurately capture facial surfaces, making it possible for precise description of facial activities. In this paper, we present a novel mesh-based method for 3D facial expression recognition using two local shape descriptors. To characterize shape information of the local neighborhood of facial landmarks, we calculate the weighted statistical distributions of surface differential quantities, including histogram of mesh gradient (HoG) and histogram of shape index (HoS). Normal cycle theory based curvature estimation method is employed on 3D face models along with the common cubic fitting curvature estimation method for the purpose of comparison. Based on the basic fact that different expressions involve different local shape deformations, the SVM classifier with both linear and RBF kernels outperforms the state of the art results on the subset of the BU-3DFE database with the same experimental setting. © 2011 Springer-Verlag.

  1. Analytical expressions of the imaging and aberration coefficients of a general form surface.

    Science.gov (United States)

    Yang, Liu; Qi, Jin Wei; Bin, Zhu

    2017-12-01

    A theoretical development is presented in this paper for describing and understanding the imaging and aberrations of a general form surface. The development is based on the Taylor expansion of an arbitrary ray trace from the object reference plane to the image reference plane, which is called the base mapping of the general form surface in this paper. The base mapping is expressed as two Taylor series of the object and pupil coordinates and the imaging and aberration coefficients in the third-order scope are derived and presented as analytical expressions relevant to the optic parameters, invoking no approximations. The situation with tilted object and observing plane is also considered, and the mapping from a tilted object to a tilted observing plane is derived via simple mathematical manipulations based on the base mapping.

  2. Systemic alteration of cell-surface and secreted glycoprotein expression in malignant breast cancer cell lines.

    Science.gov (United States)

    Timpe, Leslie C; Yen, Roger; Haste, Nicole V; Litsakos-Cheung, Christina; Yen, Ten-Yang; Macher, Bruce A

    2013-11-01

    Breast cancer cell lines express fewer transmembrane and secreted glycoproteins than nonmalignant ones. The objective of these experiments was to characterize the changes in the expression of several hundred glycoproteins quantitatively. Secreted and cell-surface glycoproteins were isolated using a glycoprotein capture protocol and then identified by tandem mass spectrometry. Glycoproteins expressed by a group of cell lines originating from malignant tumors of the breast were compared with those expressed by a nonmalignant set. The average number of spectral counts (proportional to relative protein abundance) and the total number of glycopeptides in the malignant samples were reduced to about two-thirds of the level in the nonmalignant samples. Most glycoproteins were expressed at a different level in the malignant samples, with nearly as many increasing as decreasing. The glycoproteins with reduced expression accounted for a larger change in spectral counts, and hence for the net loss of spectral counts in the malignant lines. Similar results were found when the glycoproteins were studied via identified glycosylation sites only, or through identified sites together with non-glycopeptides. The overall reduction is largely due to the loss of integrins, laminins and other proteins that form or interact with the basement membrane.

  3. Cell-surface expression of Hsp70 on hematopoietic cancer cells after inhibition of HDAC activity

    DEFF Research Database (Denmark)

    Jensen, Helle

    frequently express Hsp70 on their cell surface, whereas the corresponding normal tissues do not. In addition, several clinically applied reagents, such as alkyl-lysophospholipides, chemotherapeutic agents, and anti-inflammatory reagents, have been found to enhance Hsp70 cell surface expression on cancer...

  4. Predicting the minimum liquid surface tension activity of pseudomonads expressing biosurfactants.

    Science.gov (United States)

    Mohammed, I U; Deeni, Y; Hapca, S M; McLaughlin, K; Spiers, A J

    2015-01-01

    Bacteria produce a variety of biosurfactants capable of significantly reducing liquid (aqueous) surface tension (γ) with a range of biological roles and biotechnological uses. To determine the lowest achievable surface tension (γMin ), we tested a diverse collection of Pseudomonas-like isolates from contaminated soil and activated sludge and identified those expressing biosurfactants by drop-collapse assay. Liquid surface tension-reducing ability was quantitatively determined by tensiometry, with 57 isolates found to significantly lower culture supernatant surface tensions to 24·5-49·1 mN m(-1) . Differences in biosurfactant behaviour determined by foaming, emulsion and oil-displacement assays were also observed amongst isolates producing surface tensions of 25-27 mN m(-1) , suggesting that a range of structurally diverse biosurfactants were being expressed. Individual distribution identification (IDI) analysis was used to identify the theoretical probability distribution that best fitted the surface tension data, which predicted a γMin of 24·24 mN m(-1) . This was in agreement with predictions based on earlier work of published mixed bacterial spp. data, suggesting a fundamental limit to the ability of bacterial biosurfactants to reduce surface tensions in aqueous systems. This implies a biological restriction on the synthesis and export of these agents or a physical-chemical restriction on their functioning once produced. Numerous surveys of biosurfactant-producing bacteria have been conducted, but only recently has an attempt been made to predict the minimum liquid surface tension these surface-active agents can achieve. Here, we determine a theoretical minimum of 24 mN m(-1) by statistical analysis of tensiometry data, suggesting a fundamental limit for biosurfactant activity in bacterial cultures incubated under standard growth conditions. This raises a challenge to our understanding of biosurfactant expression, secretion and function, as well as

  5. Different Culture Media Affect Proliferation, Surface Epitope Expression, and Differentiation of Ovine MSC

    Directory of Open Access Journals (Sweden)

    Carina Adamzyk

    2013-01-01

    Full Text Available Orthopedic implants including engineered bone tissue are commonly tested in sheep. To avoid rejection of heterologous or xenogeneic cells, autologous cells are preferably used, that is, ovine mesenchymal stem cells (oMSC. Unlike human MSC, ovine MSC are not well studied regarding isolation, expansion, and characterization. Here we investigated the impact of culture media composition on growth characteristics, differentiation, and surface antigen expression of oMSC. The culture media varied in fetal calf serum (FCS content and in the addition of supplements and/or additional epidermal growth factor (EGF. We found that FCS strongly influenced oMSC proliferation and that specific combinations of supplemental factors (MCDB-201, ITS-plus, dexamethasone, and L-ascorbic acid determined the expression of surface epitopes. We compared two published protocols for oMSC differentiation towards the osteogenic, adipogenic, and chondrogenic fate and found (i considerable donor to donor variations, (ii protocol-dependent variations, and (iii variations resulting from the preculture medium composition. Our results indicate that the isolation and culture of oMSC in different growth media are highly variable regarding oMSC phenotype and behaviour. Furthermore, variations from donor to donor critically influence growth rate, surface marker expression, and differentiation.

  6. Orf virus interferes with MHC class I surface expression by targeting vesicular transport and Golgi

    Directory of Open Access Journals (Sweden)

    Rohde Jörg

    2012-07-01

    Full Text Available Abstract Background The Orf virus (ORFV, a zoonotic Parapoxvirus, causes pustular skin lesions in small ruminants (goat and sheep. Intriguingly, ORFV can repeatedly infect its host, despite the induction of a specific immunity. These immune modulating and immune evading properties are still unexplained. Results Here, we describe that ORFV infection of permissive cells impairs the intracellular transport of MHC class I molecules (MHC I as a result of structural disruption and fragmentation of the Golgi apparatus. Depending on the duration of infection, we observed a pronounced co-localization of MHC I and COP-I vesicular structures as well as a reduction of MHC I surface expression of up to 50%. These subversion processes are associated with early ORFV gene expression and are accompanied by disturbed carbohydrate trimming of post-ER MHC I. The MHC I population remaining on the cell surface shows an extended half-life, an effect that might be partially controlled also by late ORFV genes. Conclusions The presented data demonstrate that ORFV down-regulates MHC I surface expression in infected cells by targeting the late vesicular export machinery and the structure and function of the Golgi apparatus, which might aid to escape cellular immune recognition.

  7. Kv7.1 surface expression is regulated by epithelial cell polarization

    DEFF Research Database (Denmark)

    Andersen, Martin N; Olesen, Søren-Peter; Rasmussen, Hanne Borger

    2011-01-01

    The potassium channel K(V)7.1 is expressed in the heart where it contributes to the repolarization of the cardiac action potential. In addition, K(V)7.1 is expressed in epithelial tissues where it plays a role in salt and water transport. Mutations in the kcnq1 gene can lead to long QT syndrome...... and deafness, and several mutations have been described as trafficking mutations. To learn more about the basic mechanisms that regulate K(V)7.1 surface expression, we have investigated the trafficking of K(V)7.1 during the polarization process of the epithelial cell line Madin-Darby Canine Kidney (MDCK) using...... is regulated by signaling mechanisms involved in epithelial cell polarization in particular signaling cascades involving protein kinase C and PI3K....

  8. Altered expression of epithelial cell surface glycoconjugates and intermediate filaments at the margins of mucosal wounds

    DEFF Research Database (Denmark)

    Dabelsteen, Erik; Grøn, B.; Mandel, U.

    1998-01-01

    Alterations in cell to cell adhesion are necessary to enable the type of cell movements that are associated with epithelial wound healing and malignant invasion. Several studies of transformed cells have related epithelial cell movement to changes in the cell surface expression of the carbohydrate......-T antigen. The changes induced by wounding in the expression of collagen IV, laminin gamma2-chain (laminin-5), and laminin alpha5-chain were similar to those found in skin wounds and served to define the region of epithelial movement. This region was found to show a marked increase in staining for both...... epithelium, a pattern of expression similar to K16, which was also strongly upregulated in both the outgrowth and the adjacent nonwounded epithelium. These findings provide further support for an influence of such carbohydrate structures on the migratory behavior of epithelial cells....

  9. Repeated exposure to morphine alters surface expression of AMPA receptors in the rat medial prefrontal cortex.

    Science.gov (United States)

    Mickiewicz, Amanda L; Napier, T Celeste

    2011-01-01

    Behavioral sensitization describes the intensification of motor activity that results from repeated exposure to drugs of misuse, and the underlying neuronal adaptations are hypothesized to model aspects of the brain changes that occur in humans misusing such drugs. The α-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate (AMPA) receptor is an ionotropic glutamate receptor involved in the neuroplasticity that accompanies acute and repeated drug administration. Changing surface expression is one means to regulate AMPA receptor function, and the present study tested the hypothesis that behavioral sensitization to the μ-opioid receptor agonist morphine is accompanied by changes in the subcellular distribution of AMPA receptors in limbic brain regions. To test this hypothesis, we used a protein cross-linking assay to assess cell surface and intracellular levels of GluA1 and GluA2 subunits in the nucleus accumbens, medial prefrontal cortex and ventral pallidum. Repeated morphine treatment decreased surface expression of GluA1 in the medial prefrontal cortex without affecting levels of GluA2. In contrast, surface levels of GluA1 or GluA2 were unchanged in the nucleus accumbens and ventral pallidum, demonstrating that although AMPA receptors in accumbal and pallidal regions are critical mediators of behaviors induced by repeated opiate exposure, these effects are not accompanied by changes in surface expression. The findings reveal that the involvement of AMPA receptor trafficking in opiate-induced behavioral sensitization is relegated to selective regions and that AMPA receptors in the medial prefrontal cortex may be particularly sensitive to these actions. © 2010 The Authors. European Journal of Neuroscience © 2010 Federation of European Neuroscience Societies and Blackwell Publishing Ltd.

  10. Dynamic expression of cell surface hydrophobicity during initial yeast cell growth and before germ tube formation of Candida albicans.

    OpenAIRE

    Hazen, B W; Hazen, K C

    1988-01-01

    Expression of cell surface hydrophobicity (CSH) during initial growth of Candida albicans was monitored. CSH of hydrophobic and hydrophilic yeast cells changed within 30 min upon subculture into fresh medium. Morphologic evidence of germination was preceded by expression of CSH. These results indicate that CSH expression is important in C. albicans growth.

  11. Salmonella regulates polyubiquitination and surface expression of MHC class II antigens.

    Science.gov (United States)

    Lapaque, Nicolas; Hutchinson, James L; Jones, Des C; Méresse, Stéphane; Holden, David W; Trowsdale, John; Kelly, Adrian P

    2009-08-18

    Salmonella typhimurium is a facultative pathogen capable of entering and replicating in both professional and non-professional antigen presenting cells. Control of infection requires MHC class II restricted CD4 T-helper cell responses. Here we show that Salmonella infection induced polyubiquitination of HLA-DR, a post-translational modification that led to removal of mature, peptide loaded, alphabeta dimers from the cell surface. Immature alphabetaIi complexes were unaffected. Surface expression of all class II isotypes, HLA-DP, -DQ, and -DR, was reduced in infected cells, but other cell-surface molecules that traffic through class II peptide loading compartments were unaffected. A Salmonella strain carrying a mutation in ssaV did not induce ubiquitination of class II, implicating Salmonella T3SS-2 effector proteins in the process. T3SS-2 effectors, with established or proposed roles in ubiquitination, were not required for class II down-regulation, suggesting that an additional T3SS-2 effector is involved in regulating MHC class II ubiquitination. Although recognized as a viral immune evasion strategy, here, we demonstrate that bacteria can control surface MHC expression through ubiquitination.

  12. Signal peptide cleavage is essential for surface expression of a regulatory T cell surface protein, leucine rich repeat containing 32 (LRRC32

    Directory of Open Access Journals (Sweden)

    Sugiyama Hideaki

    2011-05-01

    Full Text Available Abstract Background Elevated numbers of regulatory T cells (Tregs have been implicated in certain cancers. Depletion of Tregs has been shown to increase anti-tumor immunity. Tregs also play a critical role in the suppression of autoimmune responses. The study of Tregs has been hampered by a lack of adequate surface markers. Leucine Rich Repeat Containing 32 (LRRC32, also known as Glycoprotein A Repetitions Predominant (GARP, has been postulated as a novel surface marker of activated Tregs. However, there is limited information regarding the processing of LRRC32 or the regulatory phenotype and functional activity of Tregs expressing LRRC32. Results Using naturally-occurring freshly isolated Tregs, we demonstrate that low levels of LRRC32 are present intracellularly prior to activation and that freshly isolated LRRC32+ Tregs are distinct from LRRC32- Tregs with respect to the expression of surface CD62L. Using LRRC32 transfectants of HEK cells, we demonstrate that the N-terminus of LRRC32 is cleaved prior to expression of the protein at the cell surface. Furthermore, we demonstrate using a construct containing a deleted putative signal peptide region that the presence of a signal peptide region is critical to cell surface expression of LRRC32. Finally, mixed lymphocyte assays demonstrate that LRRC32+ Tregs are more potent suppressors than LRRC32- Tregs. Conclusions A cleaved signal peptide site in LRRC32 is necessary for surface localization of native LRRC32 following activation of naturally-occurring freshly-isolated regulatory T cells. LRRC32 expression appears to alter the surface expression of activation markers of T cells such as CD62L. LRRC32 surface expression may be useful as a marker that selects for more potent Treg populations. In summary, understanding the processing and expression of LRRC32 may provide insight into the mechanism of action of Tregs and the refinement of immunotherapeutic strategies aimed at targeting these cells.

  13. Relationship of CD86 surface marker expression and cytotoxicity on dendritic cells exposed to chemical allergen

    International Nuclear Information System (INIS)

    Hulette, Ben C.; Ryan, Cindy A.; Gildea, Lucy A.; Gerberick, G. Frank

    2005-01-01

    Human peripheral blood-derived dendritic cells (DC) respond to a variety of chemical allergens by up-regulating expression of the co-stimulatory molecule CD86. It has been postulated that this measure might provide the basis for an in vitro alternative approach for the identification of skin sensitizing chemicals. We recently reported that DC, exposed in culture to the highest non-cytotoxic concentrations of various chemical allergens, displayed marginal up-regulation of membrane CD86 expression; the interpretation being that such changes were insufficiently sensitive for the purposes of hazard identification. For the work presented here, immature DC were derived from human monocytes and treated with the chemical allergens 2,4-dinitrobenzenesulfonic acid (DNBS), nickel sulfate (NiSO 4 ), p-phenylenediamine (PPD), Bandrowski's base (BB), hydroquinone (HQ) and propyl gallate (PG) for 48 h at concentrations which induced both no to slight to moderate cytotoxicity. For comparison, DC were treated with the irritants sodium dodecyl sulfate (SDS), benzoic acid (BA), and benzalkonium chloride (BZC) at concentrations resulting in comparable levels of cytotoxicity. CD86 expression, as measured by flow cytometry, was consistently up-regulated (ranging from 162 to 386% control) on DC treated with concentrations of chemical allergens that induced approximately 10-15% cytotoxicity. The irritants BA and BZC did not induce up-regulation of CD86 expression when tested at concentrations that induced similar levels of cytotoxicity. SDS, however, up-regulated CD86 expression to 125-138% of control in 2/4 preparations when tested at concentrations which induced similar toxicity. Our results confirm that chemical allergens up-regulate CD86 expression on blood-derived DC and illustrate further that up-regulation of CD86 surface marker expression is more robust when DC are treated with concentrations of chemical allergen that induce slight to moderate cytotoxicity

  14. Bacillus Cellulase Molecular Cloning, Expression, and Surface Display on the Outer Membrane of Escherichia coli

    Directory of Open Access Journals (Sweden)

    Daehwan Kim

    2018-02-01

    Full Text Available One of the main challenges of using recombinant enzymes is that they are derived from genetically-modified microorganisms commonly located in the intracellular region. The use of these recombinant enzymes for commercial purposes requires the additional processes of cell disruption and purification, which may result in enzyme loss, denaturation, and increased total production cost. In this study, the cellulase gene of Bacillus licheniformis ATCC 14580 was cloned, over-expressed, and surface displayed in recombinant Escherichia coli using an ice-nucleation protein (INP. INP, an outer membrane-bound protein from Pseudomonas syringae, was utilized as an anchor linker, which was cloned with a foreign cellulase gene into the pET21a vector to develop a surface display system on the outer membrane of E. coli. The resulting strain successfully revealed cellulase on the host cell surface. The over-expressed INP-cellulase fusion protein was confirmed via staining assay for determining the extracellular cellulase and Western blotting method for the molecular weight (MW of cellulase, which was estimated to be around 61.7 kDa. Cell fractionation and localization tests demonstrated that the INP-cellulase fusion protein was mostly present in the supernatant (47.5% and outer membrane (19.4%, while the wild-type strain intracellularly retained enzymes within cytosol (>61%, indicating that the INP gene directed the cellulase expression on the bacteria cell surface. Further studies of the optimal enzyme activity were observed at 60 °C and pH 7.0, and at least 75% of maximal enzyme activity was preserved at 70 °C.

  15. TGF-β induces surface LAP expression on murine CD4 T cells independent of Foxp3 induction.

    Directory of Open Access Journals (Sweden)

    Takatoku Oida

    2010-11-01

    Full Text Available It has been reported that human FOXP3(+ CD4 Tregs express GARP-anchored surface latency-associated peptide (LAP after activation, based on the use of an anti-human LAP mAb. Murine CD4 Foxp3(+ Tregs have also been reported to express surface LAP, but these studies have been hampered by the lack of suitable anti-mouse LAP mAbs.We generated anti-mouse LAP mAbs by immunizing TGF-β(-/- animals with a mouse Tgfb1-transduced P3U1 cell line. Using these antibodies, we demonstrated that murine Foxp3(+ CD4 Tregs express LAP on their surface. In addition, retroviral transduction of Foxp3 into mouse CD4(+CD25(- T cells induced surface LAP expression. We then examined surface LAP expression after treating CD4(+CD25(- T cells with TGF-β and found that TGF-β induced surface LAP not only on T cells that became Foxp3(+ but also on T cells that remained Foxp3(- after TGF-β treatment. GARP expression correlated with the surface LAP expression, suggesting that surface LAP is GARP-anchored also in murine T cells.Unlike human CD4 T cells, surface LAP expression on mouse CD4 T cells is controlled by Foxp3 and TGF-β. Our newly described anti-mouse LAP mAbs will provide a useful tool for the investigation and functional analysis of T cells that express LAP on their surface.

  16. The Cell Surface Markers Expression in Postmenopausal Women and Relation to Obesity and Bone Status

    Directory of Open Access Journals (Sweden)

    Mira Horváthová

    2017-07-01

    Full Text Available The age-related changes and hormonal deprivation in postmenopausal women are associated with the immune response alteration. The excessive fat accumulation, local and systemic inflammation may lead to dysregulation in immune function and relevant health problems, including obesity and osteoporosis. We analyzed the expression of cell surface markers in the venous blood specimens, stained with fluorophores-conjugated monoclonal antibodies and analysed by multicolour flow cytometry. The significant changes of cytotoxic, naive, and memory T-lymphocytes, plasmacytoid dendritic cells (DCs were in postmenopausal women versus fertile women. Body mass index (BMI affected markedly the cell surface expression of CD265/RANK. Osteoporosis is linked to reduced percentage of plasmacytoid DCs, and elevated natural Treg cells (p < 0.05. The confounding factors such as women age, BMI, bone mineral density (BMD, waist size and tissue fat affect the expression of RANK on myeloid DCs and CD40L on T-lymphocytes that might be the immunophenotypic modulators after menopause.

  17. Biofilm-Forming Staphylococcus epidermidis Expressing Vancomycin Resistance Early after Adhesion to a Metal Surface

    Directory of Open Access Journals (Sweden)

    Toshiyuki Sakimura

    2015-01-01

    Full Text Available We investigated biofilm formation and time of vancomycin (VCM resistance expression after adhesion to a metal surface in Staphylococcus epidermidis. Biofilm-forming Staphylococcus epidermidis with a VCM MIC of 1 μg/mL was used. The bacteria were made to adhere to a stainless steel washer and treated with VCM at different times and concentrations. VCM was administered 0, 2, 4, and 8 hours after adhesion. The amount of biofilm formed was evaluated based on the biofilm coverage rates (BCRs before and after VCM administration, bacterial viability in biofilm was visually observed using the fluorescence staining method, and the viable bacterial count in biofilm was measured. The VCM concentration required to decrease BCR significantly compared with that of VCM-untreated bacteria was 4 μg/mL, even in the 0 hr group. In the 4 and 8 hr groups, VCM could not inhibit biofilm growth even at 1,024 μg/mL. In the 8 hr group, viable bacteria remained in biofilm at a count of 104 CFU even at a high VCM concentration (1,024 μg/mL. It was suggested that biofilm-forming Staphylococcus epidermidis expresses resistance to VCM early after adhesion to a metal surface. Resistance increased over time after adhesion as the biofilm formed, and strong resistance was expressed 4–8 hours after adhesion.

  18. Regulation of NKG2D-ligand cell surface expression by intracellular calcium after HDAC-inhibitor treatment

    DEFF Research Database (Denmark)

    Jensen, Helle; Hagemann-Jensen, Michael Henrik; Lauridsen, Felicia Kathrine Bratt

    2013-01-01

    cell surface expression on melanoma cells and Jurkat T-cells. A NKG2D-dependent cytolytic assay and staining with a recombinant NKG2D-Fc fusion protein showed that calcium chelation impaired the functional ability of NKG2D-ligands induced by HDAC-inhibitor treatment. The HDAC-inhibitor induced cell...... surface expression of ULBP2, but not MICA/B, was sensitive to treatment calmidazolium and trifluoperazine, two agents known to block calcium signaling. siRNA-mediated knock-down of the calcium-regulated proteins calmodulin or calpain did however not affect NKG2D-ligand cell surface expression on Jurkat T...

  19. A universal empirical expression for the isotope surface exchange coefficients (k*) of acceptor-doped perovskite and fluorite oxides.

    Science.gov (United States)

    De Souza, R A

    2006-02-21

    The isotope surface exchange coefficient k* determined in an 18O/16O exchange experiment characterises the exchange flux of the dynamic equilibrium between oxygen in the gas phase and oxygen in a solid oxide. At present there is no atomistic expression that relates measured exchange coefficients to materials' parameters. In this study an empirical, atomistic expression is developed that describes the exchange kinetics of gaseous oxygen with diverse acceptor-doped perovskite and fluorite oxides at temperatures above T approximately 900 K. The expression is used to explain the observed correlations between surface exchange coefficients k* and oxygen tracer diffusion coefficients D* and to identify compounds that exhibit high surface exchange coefficients.

  20. Transcriptomic landscape of acute promyelocytic leukemia reveals aberrant surface expression of the platelet aggregation agonist Podoplanin.

    Science.gov (United States)

    Lavallée, Vincent-Philippe; Chagraoui, Jalila; MacRae, Tara; Marquis, Miriam; Bonnefoy, Arnaud; Krosl, Jana; Lemieux, Sébastien; Marinier, Anne; Pabst, Caroline; Rivard, Georges-Étienne; Hébert, Josée; Sauvageau, Guy

    2018-02-23

    Acute promyelocytic leukemia (APL) is a medical emergency because of associated lethal early bleeding, a condition preventable by prompt diagnosis and therapeutic intervention. The mechanisms underlying the hemostatic anomalies of APL are not completely elucidated. RNA-sequencing-based characterization of APL (n = 30) was performed and compared to that of other acute myeloid leukemia (n = 400) samples and normal promyelocytes. Perturbations in the transcriptome of coagulation and fibrinolysis-related genes in APL extend beyond known culprits and now include Thrombin, Factor X and Urokinase Receptor. Most intriguingly, the Podoplanin (PDPN) gene, involved in platelet aggregation, is aberrantly expressed in APL promyelocytes and is the most distinctive transcript for this disease. Using an antibody panel optimized for AML diagnosis by flow cytometry, we also found that PDPN was the most specific surface marker for APL, and that all-trans retinoic acid therapy rapidly decreases its expression. Functional studies showed that engineered overexpression of this gene in human leukemic cells causes aberrant platelet binding, activation and aggregation. PDPN-expressing primary APL cells, but not PDPN-negative primary leukemias, specifically induce platelet binding, activation and aggregation. Finally, PDPN expression on leukemia cells in a xenograft model was associated with thrombocytopenia and prolonged bleeding time in vivo. Together our results suggest that PDPN may contribute to the hemostatic perturbations found in APL.

  1. Investigation of integrin expression on the surface of osteoblast-like cells by atomic force microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Caneva Soumetz, Federico [Department of Communication, Computer and System Sciences, University of Genova, Via Opera Pia, 13-16145 Genova (Italy); Saenz, Jose F. [Biophysical and Electronic Engineering Department, University of Genova, Via All' Opera Pia 11a, 16145 Genova (Italy); Pastorino, Laura; Ruggiero, Carmelina [Department of Communication, Computer and System Sciences, University of Genova, Via Opera Pia, 13-16145 Genova (Italy); Nosi, Daniele [Department of Anatomy, Histology and Forensic Medicine, Bio-photonic Laboratory, University of Florence, viale Morgagni, 85 Firenze, CAP 50134 Florence (Italy); Raiteri, Roberto, E-mail: rr@unige.it [Biophysical and Electronic Engineering Department, University of Genova, Via All' Opera Pia 11a, 16145 Genova (Italy)

    2010-03-15

    The transforming growth factor {beta}1 (TGF-{beta}1) is a human cytokine which has been demonstrated to modulate cell surface integrin repertoire. In this work integrin expression in response to TGF-{beta}1 stimulation has been investigated on the surface of human osteoblast-like cells. We used atomic force microscopy (AFM) and confocal laser scanning microscopy to assess integrin expression and to evaluate their distribution over the dorsal side of the plasma membrane. AFM probes have been covalently functionalised with monoclonal antibodies specific to the {beta}1 integrin subunit. Force curves have been collected in order to obtain maps of the interaction between the immobilized antibody and the respective cell membrane receptors. Adhesion peaks have been automatically detected by means of an ad hoc developed data analysis software. The specificity of the detected interactions has been assessed by adding free antibody in the solution and monitoring the dramatic decrease in the recorded interactions. In addition, the effect of TGF-{beta}1 treatment on both the fluorescence signal and the adhesion events has been tested. The level of expression of the {beta}1 integrin subunit was enhanced by TGF-{beta}1. As a further analysis, the adhesion force of the single living cells to the substrate was measured by laterally pushing the cell with the AFM tip and measuring the force necessary to displace it. The treatment with TGF-{beta}1 resulted in a decrease of the cell/substrate adhesion force. Results obtained by AFM have been validated by confocal laser scanning microscopy thus demonstrating the high potential of the AFM technique for the investigation of cell surface receptors distribution and trafficking at the nanoscale.

  2. Investigation of integrin expression on the surface of osteoblast-like cells by atomic force microscopy

    International Nuclear Information System (INIS)

    Caneva Soumetz, Federico; Saenz, Jose F.; Pastorino, Laura; Ruggiero, Carmelina; Nosi, Daniele; Raiteri, Roberto

    2010-01-01

    The transforming growth factor β1 (TGF-β1) is a human cytokine which has been demonstrated to modulate cell surface integrin repertoire. In this work integrin expression in response to TGF-β1 stimulation has been investigated on the surface of human osteoblast-like cells. We used atomic force microscopy (AFM) and confocal laser scanning microscopy to assess integrin expression and to evaluate their distribution over the dorsal side of the plasma membrane. AFM probes have been covalently functionalised with monoclonal antibodies specific to the β1 integrin subunit. Force curves have been collected in order to obtain maps of the interaction between the immobilized antibody and the respective cell membrane receptors. Adhesion peaks have been automatically detected by means of an ad hoc developed data analysis software. The specificity of the detected interactions has been assessed by adding free antibody in the solution and monitoring the dramatic decrease in the recorded interactions. In addition, the effect of TGF-β1 treatment on both the fluorescence signal and the adhesion events has been tested. The level of expression of the β1 integrin subunit was enhanced by TGF-β1. As a further analysis, the adhesion force of the single living cells to the substrate was measured by laterally pushing the cell with the AFM tip and measuring the force necessary to displace it. The treatment with TGF-β1 resulted in a decrease of the cell/substrate adhesion force. Results obtained by AFM have been validated by confocal laser scanning microscopy thus demonstrating the high potential of the AFM technique for the investigation of cell surface receptors distribution and trafficking at the nanoscale.

  3. Inflammation promotes oral squamous carcinoma immune evasion via induced programmed death ligand-1 surface expression.

    Science.gov (United States)

    Lu, Wanlu; Lu, Libing; Feng, Yun; Chen, Jiao; Li, Yan; Kong, Xiangli; Chen, Sixiu; Li, Xiaoyu; Chen, Qianming; Zhang, Ping

    2013-05-01

    The association between inflammation and cancer provides a new target for tumor biotherapy. The inflammatory cells and molecules within the tumor microenvironment have decisive dual roles in antitumor immunity and immune evasion. In the present study, phytohemagglutinin (PHA) was used to stimulate peripheral blood mononuclear cells (PBMCs) to simulate the tumor inflammatory microenvironment. The effect of immune cells and inflammatory cytokines on the surface expression of programmed cell death-1 ligand 1 (PD-L1) and tumor immune evasion was investigated using flow cytometry (FCM) and an in vivo xenotransplantation model. Based on the data, PHA-activated, but not resting, immune cells were able to promote the surface expression of PD-L1 in Tca8113 oral squamous carcinoma cells via the secretion of inflammatory cytokines, but not by cell-cell contact. The majority of the inflammatory cytokines had no significant effect on the proliferation, cell cycle progression and apoptosis of the Tca8113 cells, although they each induced the expression of PD-L1 in a dose-dependent manner. In total, 99% of the Tca8113 cells expressed PD-L1 following treatment with the supernatant of PHA-stimulated PBMCs. The PHA-supernatant pretreated Tca8113 cells unusually induced Tca8113 antigen-specific CD8 + T cell apoptosis in vitro and the evasion of antigen-specific T cell attraction in a nude mouse tumor-bearing model. These results indicate a new mechanism for the promotion of tumor immune evasion by the tumor inflammatory microenvironment.

  4. Expressions to Rayleigh circumferential phase velocity and dispersion relation for a cylindrical surface under mechanical pressure

    Science.gov (United States)

    Sebold, Jean Eduardo; de Lacerda, Luiz Alkimin

    2018-04-01

    This paper describes a substantiated mathematical theory for Rayleigh waves propagated on some types of metal cylinders. More specifically, it presents not only a new way to express the dispersion relation of Rayleigh waves propagated on the cylindrical surface, but also how it can be used to construct a mathematical equation showing that the applied static mechanical pressure affects the shear modulus of the metal cylinder. All steps, required to conclude the process, consider the equation of motion as a function of radial and circumferential coordinates only, while the axial component can be overlooked without causing any problems. Some numerical experiments are done to illustrate the changes in the Rayleigh circumferential phase velocity in a metal cylindrical section due to static mechanical pressure around its external surface.

  5. Processes of 30–90 days sea surface temperature variability in the northern Indian Ocean during boreal summer

    Digital Repository Service at National Institute of Oceanography (India)

    Vialard, J.; Jayakumar, A.; Gnanaseelan, C.; Lengaigne, M.; Sengupta, D.; Goswami, B.N.

    1998). The model also reproduces the right range of D20 depth values (100–130 m) in the Bay of Bengal. On the other hand, the model suffers from a severe bias at the northern end of the Arabian Sea, maybe due to a bad treatment of Persian Gulf and Red... reproduces quite well the spatial pattern of these heat flux perturbations, although again with a smaller amplitude (*20% underestimation). This underestimation is largely accounted by latent heat fluxes, and is linked to the wind bias mentioned above...

  6. Observations of inner shelf cross-shore surface material transport adjacent to a coastal inlet in the northern Gulf of Mexico

    Science.gov (United States)

    Roth, Mathias K.; MacMahan, Jamie; Reniers, Ad; Özgökmen, Tamay M.; Woodall, Kate; Haus, Brian

    2017-04-01

    Motivated by the Deepwater Horizon oil spill, the Surfzone and Coastal Oil Pathways Experiment obtained Acoustic Doppler Current Profiler (ADCP) Eulerian and GPS-drifter based Lagrangian "surface" (ADCP velocities, but both differ significantly from depth-averaged subsurface pathlines. Near-surface currents are linearly correlated with wind velocities (r =0.76 in the alongshore and r =0.85 in the cross-shore) at the 95% confidence level, and are 4-7 times larger than theoretical estimates of wind and wave-driven surface flow in an un-stratified water column. Differences in near-surface flow are attributed to the presence of a buoyant river plume forced by winds from passing extratropical storms. Plume boundary fronts induce a horizontal velocity gradient where drifters deployed outside of the plume in oceanic water routinely converge, slow, and are re-directed. When the plume flows west parallel to the beach, the seaward plume boundary front acts as a coastal barrier that prevents 100% of oceanic drifters from beaching within 27 km of the inlet. As a result, small-scale, wind-driven river plumes in the northern Gulf of Mexico act as coastal barriers that prevent offshore surface pollution from washing ashore west of river inlets.

  7. Plasma FGF21 concentrations, adipose fibroblast growth factor receptor-1 and β-klotho expression decrease with fasting in northern elephant seals.

    Science.gov (United States)

    Suzuki, Miwa; Lee, Andrew Y; Vázquez-Medina, José Pablo; Viscarra, Jose A; Crocker, Daniel E; Ortiz, Rudy M

    2015-05-15

    Fibroblast growth factor (FGF)-21 is secreted from the liver, pancreas, and adipose in response to prolonged fasting/starvation to facilitate lipid and glucose metabolism. Northern elephant seals naturally fast for several months, maintaining a relatively elevated metabolic rate to satisfy their energetic requirements. Thus, to better understand the impact of prolonged food deprivation on FGF21-associated changes, we analyzed the expression of FGF21, FGF receptor-1 (FGFR1), β-klotho (KLB; a co-activator of FGFR) in adipose, and plasma FGF21, glucose and 3-hydroxybutyrate in fasted elephant seal pups. Expression of FGFR1 and KLB mRNA decreased 98% and 43%, respectively, with fasting duration. While the 80% decrease in mean adipose FGF21 mRNA expression with fasting did not reach statistical significance, it paralleled the 39% decrease in plasma FGF21 concentrations suggesting that FGF21 is suppressed with fasting in elephant seals. Data demonstrate an atypical response of FGF21 to prolonged fasting in a mammal suggesting that FGF21-mediated mechanisms have evolved differentially in elephant seals. Furthermore, the typical fasting-induced, FGF21-mediated actions such as the inhibition of lipolysis in adipose may not be required in elephant seals as part of a naturally adapted mechanism to support their unique metabolic demands during prolonged fasting. Copyright © 2015 Elsevier Inc. All rights reserved.

  8. Surface expression of Helicobacter pylori HpaA adhesion antigen on Vibrio cholerae, enhanced by co-expressed enterotoxigenic Escherichia coli fimbrial antigens.

    Science.gov (United States)

    Tobias, Joshua; Lebens, Michael; Wai, Sun Nyunt; Holmgren, Jan; Svennerholm, Ann-Mari

    2017-04-01

    Helicobacter pylori infection can cause peptic ulceration and is associated with gastric adenocarcinoma. This study aimed to construct and characterize a non-virulent Vibrio cholerae O1 strain, which grows more rapidly than H. pylori, as vector for H. pylori antigens for possible use as a vaccine strain against H. pylori. This was done by recombinant expression of the H. pylori adhesion antigen HpaA alone or, as a proof of principle, together with different colonization factor (CF) antigens of enterotoxigenic Escherichia coli (ETEC) which may enhance immune responses against HpaA. A recombinant V. cholerae strain co-expressing HpaA and a fimbrial CF antigens CFA/I or CS5, but not the non-fimbrial CF protein CS6, was shown to express larger amounts of HpaA on the surface when compared with the same V. cholerae strain expressing HpaA alone. Mutations in the CFA/I operon showed that the chaperon, possibly together with the usher, was involved in enhancing the surface expression of HpaA. Oral immunization of mice with formaldehyde-inactivated recombinant V. cholerae expressing HpaA alone or together with CFA/I induced significantly higher serum antibody responses against HpaA than mice similarly immunized with inactivated HpaA-expressing H. pylori bacteria. Our results demonstrate that a non-virulent V. cholerae strain can be engineered to allow strong surface expression of HpaA, and that the expression can be further increased by co-expressing it with ETEC fimbrial antigens. Such recombinant V. cholerae strains expressing HpaA, and possibly also other H. pylori antigens, may have the potential as oral inactivated vaccine candidates against H. pylori. Copyright © 2017 Elsevier Ltd. All rights reserved.

  9. Fasting increases the phosphorylation of AMPK and expression of sirtuin1 in muscle of adult male northern elephant seals (Mirounga angustirostris).

    Science.gov (United States)

    Lee, Debby; Martinez, Bridget; Crocker, Daniel E; Ortiz, Rudy M

    2017-02-01

    Fasting typically suppresses thyroid hormone (TH)-mediated cellular events and increases sirtuin 1 (SIRT1) activity. THs may regulate metabolism through nongenomic pathways and directly through activation of adenosine monophosphate-activated protein kinase (AMPK). Adult male elephant seals ( Mirounga angustirostris ) are active, hypermetabolic, and normothermic during their annual breeding fast, which is characterized by stable TH levels. However, the contribution of TH to maintenance of their fasting metabolism is unknown. To investigate the fasting effects on cellular TH-mediated events and its potential association with SIRT1 and AMPK, we quantified plasma TH levels, mRNA expressions of muscle SIRT1 and TH-associated genes as well as the phosphorylation of AMPK in adult, male northern elephant seals ( n  = 10/fasting period) over 8 weeks of fasting (early vs. late). Deiodinase type I (DI1) expression increased twofold with fasting duration suggesting that the potential for TH-mediated cellular signaling is increased. AMPK phosphorylation increased 61 ± 21% with fasting suggesting that cellular metabolism is increased. The mRNA expression of the TH transporter, monocarboxylate transporter 10 (MCT10), increased 2.4-fold and the TH receptor (THr β -1) decreased 30-fold suggesting that cellular uptake of T 4 is increased, but its subsequent cellular effects such as activation of AMPK are likely nongenomic. The up-regulation of SIRT1 mRNA expression (2.6-fold) likely contributes to the nongenomic activation of AMPK by TH, which may be necessary to maintain the expression of PGC-1 α These coordinated changes likely contribute to the up-regulation of mitochondrial metabolism to support the energetic demands associated with prolonged fasting in adult seals. © 2017 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on behalf of The Physiological Society and the American Physiological Society.

  10. Mendelian and non-mendelian mutations affecting surface antigen expression in Paramecium tetraurelia

    Energy Technology Data Exchange (ETDEWEB)

    Epstein, L.M.; Forney, J.D.

    1984-08-01

    A screening procedure was devised for the isolation of X-ray-induced mutations affecting the expression of the A immobilization antigen (i-antigen) in Paramecium tetraurelia. Two of the mutations isolated by this procedure proved to be in modifier genes. The two genes are unlinked to each other and unlinked to the structural A i-antigen gene. These are the first modifier genes identified in a Paramecium sp. that affect surface antigen expression. Another mutation was found to be a deletion of sequences just downstream from the A i-antigen gene. In cells carrying this mutation, the A i-antigen gene lies in close proximity to the end of a macronuclear chromosome. The expression of the A i-antigen is not affected in these cells, demonstrating that downstream sequences are not important for the regulation and expression of the A i-antigen gene. A stable cell line was also recovered which shows non-Mendelian inheritance of a macronuclear deletion of the A i-antigen gene. This mutant does not contain the gene in its macronucleus, but contains a complete copy of the gene in its micronucleus. In the cytoplasm of wild-type animals, the micronuclear gene is included in the developing macronucleus; in the cytoplasm of the mutant, the incorporation of the A i-antigen gene into the macronucleus is inhibited. This is the first evidence that a mechanism is available in ciliates to control the expression of a gene by regulating its incorporation into developing macronuclei.

  11. Mendelian and non-mendelian mutations affecting surface antigen expression in Paramecium tetraurelia

    International Nuclear Information System (INIS)

    Epstein, L.M.; Forney, J.D.

    1984-01-01

    A screening procedure was devised for the isolation of X-ray-induced mutations affecting the expression of the A immobilization antigen (i-antigen) in Paramecium tetraurelia. Two of the mutations isolated by this procedure proved to be in modifier genes. The two genes are unlinked to each other and unlinked to the structural A i-antigen gene. These are the first modifier genes identified in a Paramecium sp. that affect surface antigen expression. Another mutation was found to be a deletion of sequences just downstream from the A i-antigen gene. In cells carrying this mutation, the A i-antigen gene lies in close proximity to the end of a macronuclear chromosome. The expression of the A i-antigen is not affected in these cells, demonstrating that downstream sequences are not important for the regulation and expression of the A i-antigen gene. A stable cell line was also recovered which shows non-Mendelian inheritance of a macronuclear deletion of the A i-antigen gene. This mutant does not contain the gene in its macronucleus, but contains a complete copy of the gene in its micronucleus. In the cytoplasm of wild-type animals, the micronuclear gene is included in the developing macronucleus; in the cytoplasm of the mutant, the incorporation of the A i-antigen gene into the macronucleus is inhibited. This is the first evidence that a mechanism is available in ciliates to control the expression of a gene by regulating its incorporation into developing macronuclei

  12. The brain-specific Beta4 subunit downregulates BK channel cell surface expression.

    Directory of Open Access Journals (Sweden)

    Sonal Shruti

    Full Text Available The large-conductance K(+ channel (BK channel can control neural excitability, and enhanced channel currents facilitate high firing rates in cortical neurons. The brain-specific auxiliary subunit β4 alters channel Ca(++- and voltage-sensitivity, and β4 knock-out animals exhibit spontaneous seizures. Here we investigate β4's effect on BK channel trafficking to the plasma membrane. Using a novel genetic tag to track the cellular location of the pore-forming BKα subunit in living cells, we find that β4 expression profoundly reduces surface localization of BK channels via a C-terminal ER retention sequence. In hippocampal CA3 neurons from C57BL/6 mice with endogenously high β4 expression, whole-cell BK channel currents display none of the characteristic properties of BKα+β4 channels observed in heterologous cells. Finally, β4 knock-out animals exhibit a 2.5-fold increase in whole-cell BK channel current, indicating that β4 also regulates current magnitude in vivo. Thus, we propose that a major function of the brain-specific β4 subunit in CA3 neurons is control of surface trafficking.

  13. Effect of flagella expression on adhesion of Achromobacter piechaudii to chalk surfaces.

    Science.gov (United States)

    Nejidat, A; Saadi, I; Ronen, Z

    2008-12-01

    To examine flagella role and cell motility in adhesion of Achromobacter piechaudii to chalk. Transmission electron microscopy revealed that stationary cells have thicker and longer flagella than logarithmic cells. SDS-PAGE analysis showed that flagellin was more abundant in stationary cells than logarithmic ones. Sonication or inhibition of flagellin synthesis caused a 30% reduction in adhesion to chalk. Preincubation of chalk with flagella extracts reduced adhesion, by 50%. Three motility mutants were isolated. Mutants 94 and 153 were nonmotile, expressed normal levels of flagellin, have regular flagella and exhibited reduced adhesion. Mutant 208 expressed low levels of flagellin, no flagella and a spherical cell shape but with normal adhesion capacity. Multiple cell surface factors affect the adhesion efficiency to chalk. Flagella per se through physical interaction and through cell motility contribute to the adhesion process. The adhesion behaviour of mutant 208 suggests that cell shape can compensate for flagellar removal and motility. Physiological status affects bacterial cell surface properties and hence adhesion efficiency to chalk. This interaction is essential to sustain biodegradation activities and thus, remediation of contaminated chalk aquifers.

  14. Optimization of recombinant β-NGF expression in Escherichia coli using response surface methodology.

    Science.gov (United States)

    Gholami Tilko, Pouria; Hajihassan, Zahra; Moghimi, Hamid

    2017-04-21

    Human nerve growth factor a member of the neurotrophin family can be used to treat neurodegenerative diseases. As it has disulfide bonds in its structure, periplasmic expression of it using appropriate signal sequence is beneficial. Therefore, in this work β-nerve growth factor (β-NGF) was expressed in Escherichia coli using pET39b expression vector containing DsbA signal sequence. In an initial step, the effect of isopropyl β-D-1-thiogalactopyranoside (IPTG) and lactose concentration as inducer on protein production was investigated using response surface methodology. Then the effect of different postinduction time and temperature on protein production was studied. Our results indicated that the highest β-NGF production was achieved with 1 mM IPTG and low concentrations of lactose (0-2% w/v), low cultivation temperature of 25°C and postinduction time of 2 hr. Also following β-NGF purification, bioassay test using PC12 cell line was done. The biological activity of the purified β-NGF showed a similar cell proliferation activity with the standard recombinant human β-NGF. In conclusion, the results indicated an optimized upstream process to obtain high yields of biologically active β-NGF.

  15. Increased parasite surface antigen-2 expression in clinical isolates of Leishmania donovani augments antimony resistance.

    Science.gov (United States)

    Bhandari, Vasundhra; Kumar, Dhiraj; Verma, Sandeep; Srividya, Gurumurthy; Negi, Narendra Singh; Singh, Ruchi; Salotra, Poonam

    2013-11-01

    Resistance to sodium antimony gluconate (SAG) is a major cause of therapeutic failure in a large proportion of visceral leishmaniasis (VL) cases. Determinants of SAG resistance have been widely studied; however, the mechanism operating in clinical isolates is poorly understood. In the present study, expression of parasite surface antigen-2 (PSA-2) gene was studied in clinical isolates of Leishmania donovani comprising of antimony resistant (n=10) and sensitive (n=4) parasites. The expression of PSA-2 gene was found to be consistently high in SAG resistant clinical isolates (≥1.5-fold) at both transcript and protein level. Further, over-expression of PSA-2 in L. donovani isolates (LdPSA-2(++)) resulted in conversion of SAG sensitive phenotype to resistant. The LdPSA-2(++) parasites showed significantly decreased susceptibility towards SAG (>12-fold), amphotericin B (>4-fold) and miltefosine (>2.5-fold). Marked decrease in antimony accumulation and enhanced tolerance towards complement mediated lysis was evident in LdPSA-2(++) parasites. The study established the role of PSA-2 gene in SAG resistance and its potential as a biomarker to distinguish resistant and sensitive clinical isolates of L. donovani. Copyright © 2013 Elsevier Inc. All rights reserved.

  16. Exploring codon optimization and response surface methodology to express biologically active transmembrane RANKL in E. coli.

    Science.gov (United States)

    Maharjan, Sushila; Singh, Bijay; Bok, Jin-Duck; Kim, Jeong-In; Jiang, Tao; Cho, Chong-Su; Kang, Sang-Kee; Choi, Yun-Jaie

    2014-01-01

    Receptor activator of nuclear factor (NF)-κB ligand (RANKL), a master cytokine that drives osteoclast differentiation, activation and survival, exists in both transmembrane and extracellular forms. To date, studies on physiological role of RANKL have been mainly carried out with extracellular RANKL probably due to difficulties in achieving high level expression of functional transmembrane RANKL (mRANKL). In the present study, we took advantage of codon optimization and response surface methodology to optimize the soluble expression of mRANKL in E. coli. We optimized the codon usage of mRANKL sequence to a preferred set of codons for E. coli changing its codon adaptation index from 0.64 to 0.76, tending to increase its expression level in E. coli. Further, we utilized central composite design to predict the optimum combination of variables (cell density before induction, lactose concentration, post-induction temperature and post-induction time) for the expression of mRANKL. Finally, we investigated the effects of various experimental parameters using response surface methodology. The best combination of response variables was 0.6 OD600, 7.5 mM lactose, 26°C post-induction temperature and 5 h post-induction time that produced 52.4 mg/L of fusion mRANKL. Prior to functional analysis of the protein, we purified mRANKL to homogeneity and confirmed the existence of trimeric form of mRANKL by native gel electrophoresis and gel filtration chromatography. Further, the biological activity of mRANKL to induce osteoclast formation on RAW264.7 cells was confirmed by tartrate resistant acid phosphatase assay and quantitative real-time polymerase chain reaction assays. Importantly, a new finding from this study was that the biological activity of mRANKL is higher than its extracellular counterpart. To the best of our knowledge, this is the first time to report heterologous expression of mRANKL in soluble form and to perform a comparative study of functional properties of both

  17. Exploring codon optimization and response surface methodology to express biologically active transmembrane RANKL in E. coli.

    Directory of Open Access Journals (Sweden)

    Sushila Maharjan

    Full Text Available Receptor activator of nuclear factor (NF-κB ligand (RANKL, a master cytokine that drives osteoclast differentiation, activation and survival, exists in both transmembrane and extracellular forms. To date, studies on physiological role of RANKL have been mainly carried out with extracellular RANKL probably due to difficulties in achieving high level expression of functional transmembrane RANKL (mRANKL. In the present study, we took advantage of codon optimization and response surface methodology to optimize the soluble expression of mRANKL in E. coli. We optimized the codon usage of mRANKL sequence to a preferred set of codons for E. coli changing its codon adaptation index from 0.64 to 0.76, tending to increase its expression level in E. coli. Further, we utilized central composite design to predict the optimum combination of variables (cell density before induction, lactose concentration, post-induction temperature and post-induction time for the expression of mRANKL. Finally, we investigated the effects of various experimental parameters using response surface methodology. The best combination of response variables was 0.6 OD600, 7.5 mM lactose, 26°C post-induction temperature and 5 h post-induction time that produced 52.4 mg/L of fusion mRANKL. Prior to functional analysis of the protein, we purified mRANKL to homogeneity and confirmed the existence of trimeric form of mRANKL by native gel electrophoresis and gel filtration chromatography. Further, the biological activity of mRANKL to induce osteoclast formation on RAW264.7 cells was confirmed by tartrate resistant acid phosphatase assay and quantitative real-time polymerase chain reaction assays. Importantly, a new finding from this study was that the biological activity of mRANKL is higher than its extracellular counterpart. To the best of our knowledge, this is the first time to report heterologous expression of mRANKL in soluble form and to perform a comparative study of functional

  18. Surface expression of protein A on magnetosomes and capture of pathogenic bacteria by magnetosome/ antibody complexes

    Directory of Open Access Journals (Sweden)

    Jun eXu

    2014-04-01

    Full Text Available Magnetosomes are membrane-enclosed magnetite nanocrystals synthesized by magnetotactic bacteria (MTB. They display chemical purity, narrow size ranges, and species-specific crystal morphologies. Specific transmembrane proteins are sorted to the magnetosome membrane (MM. MamC is the most abundant MM protein of Magnetospirillum gryphiswaldense strain MSR-1. MamF is the second most abundant MM protein of MSR-1 and forms stable oligomers. We expressed staphylococcal protein A (SPA, an immunoglobulin-binding protein from the cell wall of Staphylococcus aureus, on MSR-1 magnetosomes by fusion with MamC or MamF. The resulting recombinant magnetosomes were capable of self-assembly with the Fc region of mammalian antibodies (Abs and were therefore useful for functionalization of magnetosomes. Recombinant plasmids pBBR-mamC-spa and pBBR-mamF-spa were constructed by fusing spa (the gene that encodes SPA with mamC and mamF, respectively. Recombinant magnetosomes with surface expression of SPA were generated by introduction of these fusion genes into wild-type MSR-1 or a mamF mutant strain. Studies with a Zeta Potential Analyzer showed that the recombinant magnetosomes had hydrated radii significantly smaller than those of WT magnetosomes and zeta potentials less than -30 mV, indicating that the magnetosome colloids were relatively stable. Observed conjugation efficiencies were as high as 71.24 µg Ab per mg recombinant magnetosomes, and the conjugated Abs retained most of their activity. Numbers of Vibrio parahaemolyticus (a common pathogenic bacterium in seafood captured by recombinant magnetosome/ Ab complexes were measured by real-time fluorescence-based quantitative PCR. One mg of complex was capable of capturing as many as 1.74×107 Vibrio cells. The surface expression system described here will be useful for design of functionalized magnetosomes from MSR-1 and other MTB.

  19. N-linked glycans are required on epithelial Na+ channel subunits for maturation and surface expression.

    Science.gov (United States)

    Kashlan, Ossama B; Kinlough, Carol L; Myerburg, Michael M; Shi, Shujie; Chen, Jingxin; Blobner, Brandon M; Buck, Teresa M; Brodsky, Jeffrey L; Hughey, Rebecca P; Kleyman, Thomas R

    2018-03-01

    Epithelial Na + channel (ENaC) subunits undergo N-linked glycosylation in the endoplasmic reticulum where they assemble into an αβγ complex. Six, 13, and 5 consensus sites (Asn-X-Ser/Thr) for N-glycosylation reside in the extracellular domains of the mouse α-, β-, and γ-subunits, respectively. Because the importance of ENaC N-linked glycans has not been fully addressed, we examined the effect of preventing N-glycosylation of specific subunits on channel function, expression, maturation, and folding. Heterologous expression in Xenopus oocytes or Fischer rat thyroid cells with αβγ-ENaC lacking N-linked glycans on a single subunit reduced ENaC activity as well as the inhibitory response to extracellular Na + . The lack of N-linked glycans on the β-subunit also precluded channel activation by trypsin. However, channel activation by shear stress was N-linked glycan independent, regardless of which subunit was modified. We also discovered that the lack of N-linked glycans on any one subunit reduced the total and surface levels of cognate subunits. The lack of N-linked glycans on the β-subunit had the largest effect on total levels, with the lack of N-linked glycans on the γ- and α-subunits having intermediate and modest effects, respectively. Finally, channels with wild-type β-subunits were more sensitive to limited trypsin proteolysis than channels lacking N-linked glycans on the β-subunit. Our results indicate that N-linked glycans on each subunit are required for proper folding, maturation, surface expression, and function of the channel.

  20. Spectral detection of near-surface moisture content and water-table position in northern peatland ecosystems

    Science.gov (United States)

    Karl M. Meingast; Michael J. Falkowski; Evan S. Kane; Lynette R. Potvin; Brian W. Benscoter; Alistair M.S. Smith; Laura L. Bourgeau-Chavez; Mary Ellen. Miller

    2014-01-01

    Wildland fire occurrence has been increasing in peatland ecosystems during recent decades. As such, there is a need for broadly applicable tools to detect and monitor controls on combustion such as surface peat moisture and water-table position. A field portable spectroradiometer was used to measure surface reflectance of two Sphagnum moss-dominated...

  1. Effects of Fertilization on Surface Runoff Loss of Nitrogen and Phosphorus from Mulberry in the Northern Zhejiang Plain, China

    Directory of Open Access Journals (Sweden)

    SHI Yan-ping

    2016-11-01

    Full Text Available In 2012 and 2013, the situ experimental plots in mulberry under two different kinds of treatment(control fertilization and farmer's conventional fertilization were conducted, and the runoff water in each plot were collected and tested in a period of two years to investigate the law of runoff, the regular pattern and the influential factors of nitrogen and phosphorus losses, and to study the coefficient of fertilizer losses from mulberry in northern Zhejiang plain. The results showed that the annual rainfall runoff coefficient was about 0.253 in mulberry field in northern Zhejiang plain. In those two years, TN and TP cumulative loss load in the conventional fertilization area reached 36.13 kg·hm-2 and 3.49 kg·hm-2, of which the N, P nutrient losses of fertilizer reached 6.415 kg·hm-2 and 1.090 kg·hm-2, respectively. N, P loss coefficients of fertilizer (the difference of nitrogen or phosphorus loss in the conventional fertilization area and the control area was divided by the total amount of fertilizer application were 0.744% and 3.047%. Nitrogen loss were mainly in soluble form, in which the NO3-N and NH4-N, accounting for about 38.3% and 14.4% respectively; while the phosphorus loss were in particulate form, accounting for about 68.9%. The larger amount of nitrogen and phosphorus nutrient loss, were within the first year of the fertilization period, and the P loss was more serious than N. Within a period of fertilization, the cumulative loss of nitrogen and phosphorus caused by rainfall and the occurrence times of runoff increased with power function(R2>0.95.

  2. Enteroendocrine cells are specifically marked by cell surface expression of claudin-4 in mouse small intestine.

    Directory of Open Access Journals (Sweden)

    Takahiro Nagatake

    Full Text Available Enteroendocrine cells are solitary epithelial cells scattered throughout the gastrointestinal tract and produce various types of hormones, constituting one of the largest endocrine systems in the body. The study of these rare epithelial cells has been hampered by the difficulty in isolating them because of the lack of specific cell surface markers. Here, we report that enteroendocrine cells selectively express a tight junction membrane protein, claudin-4 (Cld4, and are efficiently isolated with the use of an antibody specific for the Cld4 extracellular domain and flow cytometry. Sorted Cld4+ epithelial cells in the small intestine exclusively expressed a chromogranin A gene (Chga and other enteroendocrine cell-related genes (Ffar1, Ffar4, Gpr119, and the population was divided into two subpopulations based on the activity of binding to Ulex europaeus agglutinin-1 (UEA-1. A Cld4+UEA-1- cell population almost exclusively expressed glucose-dependent insulinotropic polypeptide gene (Gip, thus representing K cells, whereas a Cld4+UEA-1+ cell population expressed other gut hormone genes, including glucagon-like peptide 1 (Gcg, pancreatic polypeptide-like peptide with N-terminal tyrosine amide (Pyy, cholecystokinin (Cck, secretin (Sct, and tryptophan hydroxylase 1 (Tph1. In addition, we found that orally administered luminal antigens were taken up by the solitary Cld4+ cells in the small intestinal villi, raising the possibility that enteroendocrine cells might also play a role in initiation of mucosal immunity. Our results provide a useful tool for the cellular and functional characterization of enteroendocrine cells.

  3. Imbalanced expression of functional surface molecules in regulatory and effector T cells in systemic lupus erythematosus

    International Nuclear Information System (INIS)

    Mesquita Júnior, D.; Cruvinel, W.M.; Araujo, J.A.P.; Salmazi, K.C.; Kallas, E.G.; Andrade, L.E.C.

    2014-01-01

    Regulatory T (TREG) cells play an important role in maintaining immune tolerance and avoiding autoimmunity. We analyzed the expression of membrane molecules in TREG and effector T cells in systemic lupus erythematosus (SLE). TREG and effector T cells were analyzed for the expression of CTLA-4, PD1, CD28, CD95, GITR, HLA-DR, OX40, CD40L, and CD45RO in 26 patients with active disease, 31 with inactive disease, and 26 healthy controls. TREG cells were defined as CD25 +/high CD127 Ø/low FoxP3 + , and effector T cells were defined as CD25 + CD127 + FoxP3 Ø . The ratio of TREG to effector T cells expressing GITR, PD1, HLA-DR, OX40, CD40L, and CD45RO was determined in the three groups. The frequency of TREG cells was similar in patients with SLE and controls. However, SLE patients had a decreased frequency of CTLA-4 + TREG and CD28 + TREG cells and an increased frequency of CD40L + TREG cells. There was a decrease in the TREG/effector-T ratio for GITR + , HLA-DR + , OX40 + , and CD45RO + cells, and an increased ratio of TREG/effector-T CD40L + cells in patients with SLE. In addition, CD40L + TREG cell frequency correlated with the SLE disease activity index (P=0.0163). In conclusion, our findings showed several abnormalities in the expression of functionally critical surface molecules in TREG and effector T cells in SLE that may be relevant to the pathogenesis of this disease

  4. Refrigerated storage of platelets initiates changes in platelet surface marker expression and localization of intracellular proteins.

    Science.gov (United States)

    Wood, Ben; Padula, Matthew P; Marks, Denese C; Johnson, Lacey

    2016-10-01

    Platelets (PLTs) are currently stored at room temperature (22°C), which limits their shelf life, primarily due to the risk of bacterial growth. Alternatives to room temperature storage include PLT refrigeration (2-6°C), which inhibits bacterial growth, thus potentially allowing an extension of shelf life. Additionally, refrigerated PLTs appear more hemostatically active than conventional PLTs, which may be beneficial in certain clinical situations. However, the mechanisms responsible for this hemostatic function are not well characterized. The aim of this study was to assess the protein profile of refrigerated PLTs in an effort to understand these functional consequences. Buffy coat PLTs were pooled, split, and stored either at room temperature (20-24°C) or under refrigerated (2-6°C) conditions (n = 8 in each group). PLTs were assessed for changes in external receptor expression and actin filamentation using flow cytometry. Intracellular proteomic changes were assessed using two-dimensional gel electrophoresis and Western blotting. PLT refrigeration significantly reduced the abundance of glycoproteins (GPIb, GPIX, GPIIb, and GPIV) on the external membrane. However, refrigeration resulted in the increased expression of high-affinity integrins (αIIbβ3 and β1) and activation and apoptosis markers (CD62P, CD63, and phosphatidylserine). PLT refrigeration substantially altered the abundance and localization of several cytoskeletal proteins and resulted in an increase in actin filamentation, as measured by phalloidin staining. Refrigerated storage of PLTs induces significant changes in the expression and localization of both surface-expressed and intracellular proteins. Understanding these proteomic changes may help to identify the mechanisms resulting in the refrigeration-associated alterations in PLT function and clearance. © 2016 AABB.

  5. Imbalanced expression of functional surface molecules in regulatory and effector T cells in systemic lupus erythematosus

    Energy Technology Data Exchange (ETDEWEB)

    Mesquita Júnior, D. [Disciplina de Reumatologia, Departamento de Medicina, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, SP (Brazil); Cruvinel, W.M. [Disciplina de Reumatologia, Departamento de Medicina, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, SP (Brazil); Departamento de Biomedicina, Universidade Católica de Goiás, Goiânia, GO (Brazil); Araujo, J.A.P. [Disciplina de Reumatologia, Departamento de Medicina, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, SP (Brazil); Salmazi, K.C.; Kallas, E.G. [Disciplina de Imunologia Clínica e Alergia, Departamento de Clínica Médica, Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP (Brazil); Andrade, L.E.C. [Disciplina de Reumatologia, Departamento de Medicina, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, SP (Brazil)

    2014-08-22

    Regulatory T (TREG) cells play an important role in maintaining immune tolerance and avoiding autoimmunity. We analyzed the expression of membrane molecules in TREG and effector T cells in systemic lupus erythematosus (SLE). TREG and effector T cells were analyzed for the expression of CTLA-4, PD1, CD28, CD95, GITR, HLA-DR, OX40, CD40L, and CD45RO in 26 patients with active disease, 31 with inactive disease, and 26 healthy controls. TREG cells were defined as CD25{sup +/high}CD127{sup Ø/low}FoxP3{sup +}, and effector T cells were defined as CD25{sup +}CD127{sup +}FoxP3{sup Ø}. The ratio of TREG to effector T cells expressing GITR, PD1, HLA-DR, OX40, CD40L, and CD45RO was determined in the three groups. The frequency of TREG cells was similar in patients with SLE and controls. However, SLE patients had a decreased frequency of CTLA-4{sup +}TREG and CD28{sup +}TREG cells and an increased frequency of CD40L{sup +}TREG cells. There was a decrease in the TREG/effector-T ratio for GITR{sup +}, HLA-DR{sup +}, OX40{sup +}, and CD45RO{sup +} cells, and an increased ratio of TREG/effector-T CD40L{sup +} cells in patients with SLE. In addition, CD40L{sup +}TREG cell frequency correlated with the SLE disease activity index (P=0.0163). In conclusion, our findings showed several abnormalities in the expression of functionally critical surface molecules in TREG and effector T cells in SLE that may be relevant to the pathogenesis of this disease.

  6. Flumazenil decreases surface expression of α4β2δ GABAA receptors by increasing the rate of receptor internalization.

    Science.gov (United States)

    Kuver, Aarti; Smith, Sheryl S

    2016-01-01

    Increases in expression of α4βδ GABAA receptors (GABARs), triggered by fluctuations in the neurosteroid THP (3α-OH-5α[β]-pregnan-20-one), are associated with changes in mood and cognition. We tested whether α4βδ trafficking and surface expression would be altered by in vitro exposure to flumazenil, a benzodiazepine ligand which reduces α4βδ expression in vivo. We first determined that flumazenil (100 nM-100 μM, IC50=∼1 μM) acted as a negative modulator, reducing GABA (10 μM)-gated current in the presence of 100 nM THP (to increase receptor efficacy), assessed with whole cell patch clamp recordings of recombinant α4β2δ expressed in HEK-293 cells. Surface expression of recombinant α4β2δ receptors was detected using a 3XFLAG reporter at the C-terminus of α4 (α4F) using confocal immunocytochemical techniques following 48 h exposure of cells to GABA (10 μM)+THP (100 nM). Flumazenil (10 μM) decreased surface expression of α4F by ∼60%, while increasing its intracellular accumulation, after 48 h. Reduced surface expression of α4β2δ after flumazenil treatment was confirmed by decreases in the current responses to 100 nM of the GABA agonist gaboxadol. Flumazenil-induced decreases in surface expression of α4β2δ were prevented by the dynamin blocker, dynasore, and by leupeptin, which blocks lysosomal enzymes, suggesting that flumazenil is acting to increase endocytosis and lysosomal degradation of the receptor. Flumazenil increased the rate of receptor removal from the cell surface by 2-fold, assessed using botulinum toxin B to block insertion of new receptors. These findings may suggest new therapeutic strategies for regulation of α4β2δ expression using flumazenil. Copyright © 2015 Elsevier Inc. All rights reserved.

  7. Effect of UV radiation on the surface of mammalian immunocompetent cells. 1. The change in expression of some antigens and receptors of murine spleen lymphocyte surface

    Energy Technology Data Exchange (ETDEWEB)

    Krylenkov, V.A.; Malygin, A.M. (AN SSSR, Leningrad. Inst. Tsitologii)

    1982-12-01

    Short-wave (254nm) and long-wave (365 nm) UV rays (ShUS and LUV rays) induce the increase in the expression of surface markers of T lymphocytes-THETA(Thy-1) antigens and B lymphocytes-MBLA-antigens and EAS receptors when affecting mouse spleen cells in nonlethal and small lethal doses. Total cell content with T and B lymphocyte characters in an irradiated suspension exceeds even the total cell quantity in non-irradiated suspension (100%) which points to the possibility of the expression of plasmatic membrane antigens and receptors not manifested on the surface of nonirradiated lymphocytes. In the isolethal dose range (LD/sup 15/-LD/sup 28/) ShUV rays suppress and LUV rays induce further increase of THETA and MBLA antigens expression. Among B lymphocytes surface markers the MBLA antigens are more resistant to ShUV an LUV radiation as compared with the EAC receptors.

  8. Expressions of the radius and the surface tension of surface of tension in terms of the pressure distribution for nanoscale liquid threads

    International Nuclear Information System (INIS)

    Yan Hong; Wei Jiu-An; Cui Shu-Wen; Zhu Ru-Zeng

    2013-01-01

    The expressions of the radius and the surface tension of surface of tension R s and γ s in terms of the pressure distribution for nanoscale liquid threads are of great importance for molecular dynamics (MD) simulations of the interfacial phenomena of nanoscale fluids; these two basic expressions are derived in this paper. Although these expressions were derived first in the literature [Kim B G, Lee J S, Han M H, and Park S, 2006 Nanoscale and Microscale Thermophysical Engineering, 10, 283] and used widely thereafter, the derivation is wrong both in logical structure and physical thought. In view of the importance of these basic expressions, the logic and physical mistakes appearing in that derivation are pointed out. (condensed matter: structural, mechanical, and thermal properties)

  9. Efficacy of pyramided Bt proteins Cry1F, Cry1A.105, and cry2Ab2 expressed in Smartstax corn hybrids against lepidopteran insect pests in the northern United States.

    NARCIS (Netherlands)

    Rule, D.M.; Nolting, S.P.; Prasfika, P.L.; Storer, N.P.; Hopkins, B.W.; Scherder, E.J.A.; Siebert, M.W.; Hendrix, W.H.

    2014-01-01

    Commercial field corn (Zea mays L.) hybrids transformed to express some or all of the lepidopteran insect-resistant traits present in SmartStax corn hybrids were evaluated for insecticidal efficacy against a wide range of lepidopteran corn pests common to the northern United States, during 2008 to

  10. Five Year Mean Bottom to Surface Temperature Differences in the Northern Gulf of Mexico for 2005 through 2009

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — These images were created by combining the estimates of the mean bottom to surface temperature differences to produce seasonal representations for winter, spring,...

  11. Five Year Mean Sea-surface Salinity in the Northern Gulf of Mexico for 2005 through 2009

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — These images were created by combining the mean sea-surface salinity values to produce seasonal representations for winter, spring, summer and fall. Winter includes...

  12. Five Year Mean Sea-Surface Temperature in the Northern Gulf of Mexico for 2005 through 2009

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — These images were created by combining the mean sea-surface temperature values to produce seasonal representations for winter, spring, summer and fall. Winter...

  13. Pyrosequencing-based assessment of the bacteria diversity in surface and subsurface peat layers of a northern wetland, with focus on poorly studied phyla and candidate divisions.

    Directory of Open Access Journals (Sweden)

    Yulia M Serkebaeva

    Full Text Available Northern peatlands play a key role in the global carbon and water budget, but the bacterial diversity in these ecosystems remains poorly described. Here, we compared the bacterial community composition in the surface (0-5 cm depth and subsurface (45-50 cm peat layers of an acidic (pH 4.0 Sphagnum-dominated wetland, using pyrosequencing of 16S rRNA genes. The denoised sequences (37,229 reads, average length ∼430 bp were affiliated with 27 bacterial phyla and corresponded to 1,269 operational taxonomic units (OTUs determined at 97% sequence identity. Abundant OTUs were affiliated with the Acidobacteria (35.5±2.4% and 39.2±1.2% of all classified sequences in surface and subsurface peat, respectively, Alphaproteobacteria (15.9±1.7% and 25.8±1.4%, Actinobacteria (9.5±2.0% and 10.7±0.5%, Verrucomicrobia (8.5±1.4% and 0.6±0.2%, Planctomycetes (5.8±0.4% and 9.7±0.6%, Deltaproteobacteria (7.1±0.4% and 4.4%±0.3%, and Gammaproteobacteria (6.6±0.4% and 2.1±0.1%. The taxonomic patterns of the abundant OTUs were uniform across all the subsamples taken from each peat layer. In contrast, the taxonomic patterns of rare OTUs were different from those of the abundant OTUs and varied greatly among subsamples, in both surface and subsurface peat. In addition to the bacterial taxa listed above, rare OTUs represented the following groups: Armatimonadetes, Bacteroidetes, Chlamydia, Chloroflexi, Cyanobacteria, Elusimicrobia, Fibrobacteres, Firmicutes, Gemmatimonadetes, Spirochaetes, AD3, WS1, WS4, WS5, WYO, OD1, OP3, BRC1, TM6, TM7, WPS-2, and FCPU426. OTU richness was notably higher in the surface layer (882 OTUs than in the anoxic subsurface peat (483 OTUs, with only 96 OTUs common to both data sets. Most members of poorly studied phyla, such as the Acidobacteria, Verrucomicrobia, Planctomycetes and the candidate division TM6, showed a clear preference for growth in either oxic or anoxic conditions. Apparently, the bacterial communities in surface and

  14. Distribution, sources and contamination assessment of heavy metals in surface sediments of the South Yellow Sea and northern part of the East China Sea.

    Science.gov (United States)

    Lu, Jian; Li, Anchun; Huang, Peng

    2017-11-15

    Surface sediment samples collected from the South Yellow Sea and northern part of the East China Sea during spring and autumn, respectively, were analyzed for grain size, aluminum, and heavy metals (Cr, Ni, Cu, Zn, and Pb) to evaluate heavy metal levels and the contamination status. The results showed that all of the heavy metal concentrations met the standard criteria of the Chinese National Standard Criteria for Marine Sediment Quality. Both the EFs and a multivariate analysis (PCA) indicated that Cr, Ni, Cu, and Zn were mainly from natural contributions, while Pb was influenced by anthropogenic inputs, especially during autumn. The geoaccumulation index of Pb near the mouth of the Yangtze River suggested that the pollution degree in autumn was heavier than that in spring, which might be caused by the greater river discharge in summer and more heavy metal adsorption with finer grain sizes. Copyright © 2017 Elsevier Ltd. All rights reserved.

  15. El Niño-Southern Oscillation-time scale covariation of sea surface salinity and freshwater flux in the western tropical and northern subtropical Pacific

    Science.gov (United States)

    Nagano, Akira; Hasegawa, Takuya; Ueki, Iwao; Ando, Kentaro

    2017-07-01

    We examined the covariation of sea surface salinity (SSS) and freshwater flux in the western tropical and northern subtropical Pacific on the El Niño-Southern Oscillation time scale, using a canonical correlation analysis of monthly data between 2001 and 2013. The dominant covariation, i.e., the first canonical mode, has large positive and negative amplitudes in regions east of the Philippines and New Guinea, respectively, and reaches peaks in autumn to winter of El Niño years. The positive SSS anomaly east of the Philippines is advected to the Kuroshio Extension region. We found that the second canonical mode is another coupled variation with localized amplitudes of SSS under the atmospheric convergence zones in winter to spring of La Niña years. However, the negative SSS anomaly is annihilated possibly by the evaporation in the subtropical region.

  16. Effect on HIV-1 gene expression, Tat-Vpr interaction and cell apoptosis by natural variants of HIV-1 Tat exon 1 and Vpr from Northern India.

    Directory of Open Access Journals (Sweden)

    Sneh Lata

    Full Text Available BACKGROUND: Since HIV-1 Tat and Vpr genes are involved in promoter transactivation, apoptosis, etc, we carried out studies to find out nature and extent of natural variation in the two genes from seropositive patients from Northern India and determined their functional implications. METHODS: HIV-1 tat exon 1 and vpr were amplified from the genomic DNA isolated from the blood of HIV-1 infected individuals using specific primers by Polymerase Chain reaction (PCR and subjected to extensive genetic analysis (CLUSTAL W, Simplot etc. Their expression was monitored by generating myc fusion clones. Tat exon 1 and Vpr variants were co-transfected with the reporter gene construct (LTR-luc and their transactivation potential was monitored by measuring luciferase activity. Apoptosis and cell cycle analysis was done by Propidium Iodide (PI staining followed by FACS. RESULTS: Exon 1 of tat was amplified from 21 samples and vpr was amplified from 16 samples. One of the Tat exon 1 variants showed phylogenetic relatedness to subtype B & C and turned out to be a unique recombinant. Two of the Vpr variants were B/C/D recombinants. These natural variations were found to have no impact on the stability of Tat and Vpr. These variants differed in their ability to transactivate B LTR and C LTR promoters. B/C recombinant Tat showed better co-operative interaction with Vpr. B/C/D recombination in Vpr was found to have no effect on its co-operativity with Tat. Recombinant Tat (B/C induced more apoptosis than wild type B and C Tat. The B/C/D recombination in Vpr did not affect its G2 arrest induction potential but reduced its apoptosis induction ability. CONCLUSIONS: Extensive sequence and region-specific variations were observed in Tat and Vpr genes from HIV-1 infected individuals from Northern India. These variations have functional implications & therefore important for the pathogenicity of virus.

  17. Two ovine mitochondrial DNAs harboring a fifth 75/76 bp repeat motif without altered gene expression in Northern Spain.

    Science.gov (United States)

    Lopez-Oceja, A; Gamarra, D; Cardoso, S; Palencia-Madrid, L; Juste, R A; De Pancorbo, M M

    2017-03-01

    The Basque Country is home to the Latxa sheep breed, which is divided in several varieties such as Latxa Black Face (LBKF) and Latxa Blonde Face (LBLF). Mitochondrial DNA control region analysis of 174 male sheep (97 LBKF and 77 LBLF) was performed with the objective of characterizing the maternal lineages of these two varieties that are the basis to produce the cheese with Idiazabal quality label. The percentage of unique haplotypes was 77.32% in LBKF and 67.53% in LBLF. Most of the individuals were classified into B haplogroup (98.85%), while A haplogroup was much less frequent. Two Latxa individuals (one LBKF and one LBLF), both belonging to B haplogroup, displayed an additional 75/76 bp tandem repeat motif. Only 33 other sequences with this repeat motif were found among 11 061 sheep sequences included in the GenBank database. Gene expression was analyzed in peripheral blood leukocytes since the additional 75/76 bp repeat motif falls within ETAS1, a domain with a possible function in regulation of replication and transcription. The mRNA expression from four mitochondrial genes (COI, cyt b, ND1, and ND2) was analyzed in the two individuals of this study with a fifth repeat motif and in four without it. Although lower transcription was observed when the additional 75/76 bp repeat motif was present, no statistically significant differences were observed. Therefore, the variation in the number of the 75/76 repeat motif does not seem to modify the gene expression rate in mitochondrial genes. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. Using surface creep rate to infer fraction locked for sections of the San Andreas fault system in northern California from alignment array and GPS data

    Science.gov (United States)

    Lienkaemper, James J.; McFarland, Forrest S.; Simpson, Robert W.; Caskey, S. John

    2014-01-01

    Surface creep rate, observed along five branches of the dextral San Andreas fault system in northern California, varies considerably from one section to the next, indicating that so too may the depth at which the faults are locked. We model locking on 29 fault sections using each section’s mean long‐term creep rate and the consensus values of fault width and geologic slip rate. Surface creep rate observations from 111 short‐range alignment and trilateration arrays and 48 near‐fault, Global Positioning System station pairs are used to estimate depth of creep, assuming an elastic half‐space model and adjusting depth of creep iteratively by trial and error to match the creep observations along fault sections. Fault sections are delineated either by geometric discontinuities between them or by distinctly different creeping behaviors. We remove transient rate changes associated with five large (M≥5.5) regional earthquakes. Estimates of fraction locked, the ratio of moment accumulation rate to loading rate, on each section of the fault system provide a uniform means to inform source parameters relevant to seismic‐hazard assessment. From its mean creep rates, we infer the main branch (the San Andreas fault) ranges from only 20%±10% locked on its central creeping section to 99%–100% on the north coast. From mean accumulation rates, we infer that four urban faults appear to have accumulated enough seismic moment to produce major earthquakes: the northern Calaveras (M 6.8), Hayward (M 6.8), Rodgers Creek (M 7.1), and Green Valley (M 7.1). The latter three faults are nearing or past their mean recurrence interval.

  19. The surface expression of Inertial Oscillations in the south shore of Oahu, Hawaii

    Science.gov (United States)

    Castillo-Trujillo, A. C.; Flament, P. J.

    2016-12-01

    HF radars are used to analyze the surface expression of Near Inertial Oscillations (NIO) on the south shore of Oahu, Hawaii over a two year period. The spatial coverage and high resolution of HF radars allows us to focus on the spatial structure of the NIOs in relation to the wind forcing and background circulation. Preliminary results show larger NIOs amplitudes in spring months when sporadic southwest wind events are found. These NIOs mostly display a red frequency shift, often associated with anticylonic background vorticity. The characteristics of the observed NIOs will be compared to simulations of the classic slab layer model. We will also present the energy exchange between these NIOs and the background flow.

  20. VAR2CSA expression on the surface of placenta-derived Plasmodium falciparum-infected erythrocytes

    DEFF Research Database (Denmark)

    Magistrado, Pamela; Salanti, Ali; Tuikue Ndam, Nicaise G

    2008-01-01

    intervillous space and parasite antigens. Both placental and chondroitin sulphate A-selected parasites have high-level transcripts of a unique var gene named var2csa. However, VAR2CSA has not been consistently found by proteomic analysis of placental parasites. Contrary to this, we found VAR2CSA expressed...... on the surface of infected erythrocytes from placenta. Importantly, this was achieved with cross-reactive antibodies against VAR2CSA.......Malaria remains a major threat, in sub-Saharan Africa primarily, and the most deadly infections are those with Plasmodium falciparum. Pregnancy-associated malaria is a clinically important complication of infection; it results from a unique interaction between proteoglycans in the placental...

  1. Interrelationship between chromosome 8 aneuploidy, C-MYC amplification and increased expression in individuals from northern Brazil with gastric adenocarcinoma

    Science.gov (United States)

    Calcagno, Danielle Queiroz; Leal, Mariana Ferreira; Seabra, Aline Damaceno; Khayat, André Salim; Chen, Elizabeth Suchi; Demachki, Samia; Assumpção, Paulo Pimentel; Faria, Mario Henrique Girão; Rabenhorst, Silvia Helena Barem; Ferreira, Márcia Valéria Pitombeira; Smith, Marília de Arruda Cardoso; Burbano, Rommel Rodríguez

    2006-01-01

    AIM: To investigate chromosome 8 numerical aberrations, C-MYC oncogene alterations and its expression in gastric cancer and to correlate these findings with histopathological characteristics of gastric tumors. METHODS: Specimens were collected surgically from seven patients with gastric adenocarcinomas. Immunostaining for C-MYC and dual-color fluorescence in situ hybridization (FISH) for C-MYC gene and chromosome 8 centromere were performed. RESULTS: All the cases showed chromosome 8 aneuploidy and C-MYC amplification, in both the diffuse and intestinal histopathological types of Lauren. No significant difference (P < 0.05) was observed between the level of chromosome 8 ploidy and the site, stage or histological type of the adenocarcinomas. C-MYC high amplification, like homogeneously stained regions (HSRs) and double minutes (DMs), was observed only in the intestinal-type. Structural rearrangement of C-MYC, like translocation, was observed only in the diffuse type. Regarding C-MYC gene, a significant difference (P < 0.05) was observed between the two histological types. The C-MYC protein was expressed in all the studied cases. In the intestinal-type the C-MYC immunoreactivity was localized only in the nucleus and in the diffuse type in the nucleus and cytoplasm. CONCLUSION: Distinct patterns of alterations between intestinal and diffuse types of gastric tumors support the hypothesis that these types follow different genetic pathways. PMID:17036397

  2. [Assessment of heavy metal pollution in surface sediments of rivers in northern area of Haihe River Basin, China].

    Science.gov (United States)

    Shang, Lin-Yuan; Sun, Ran-Hao; Wang, Zhao-Ming; Ji, Yu-He; Chen, Li-Ding

    2012-02-01

    Using Håkanson potential ecological risk index, the paper assesses heavy metal risk levels in northern parts of Haihe River basin based on 39 sampling sites. The results indicate that, the concentrations of Cu, Zn, Cd, and Cr in Haihe River basin are higher than the background values of heavy metals in China mainland, while the concentration of Pb is close to the background value in China mainland. Based on the potential ecological risk index for single heavy metal, the risk of Cu, Pb, Zn and Cr belongs to the "slight" level, while Cd has various risk levels at different sampling sites. Generally, the risk order of the heavy metals is Cd > Pb > Cu > Cr > Zn. According to Håkanson potential ecological risk index, 32 monitoring sites belong to "slight" level, 5 sampling sites belong to "middle" level, and 2 monitoring sites belong to "very strong" level. The most polluted sites are Tang River and Dashi River of Beijing, Juma River in Baoding. Therefore, these rivers should be taken more considerations in the river management.

  3. Campylobacter jejuni cocultured with epithelial cells reduces surface capsular polysaccharide expression.

    LENUS (Irish Health Repository)

    Corcionivoschi, N

    2012-02-01

    The host cell environment can alter bacterial pathogenicity. We employed a combination of cellular and molecular techniques to study the expression of Campylobacter jejuni polysaccharides cocultured with HCT-8 epithelial cells. After two passages, the amount of membrane-bound high-molecular-weight polysaccharide was considerably reduced. Microarray profiling confirmed significant downregulation of capsular polysaccharide (CPS) locus genes. Experiments using conditioned media showed that sugar depletion occurred only when the bacterial and epithelial cells were cocultured. CPS depletion occurred when C. jejuni organisms were exposed to conditioned media from a different C. jejuni strain but not when exposed to conditioned media from other bacterial species. Proteinase K or heat treatment of conditioned media under coculture conditions abrogated the effect on the sugars, as did formaldehyde fixation and cycloheximide treatment of host cells or chloramphenicol treatment of the bacteria. However, sugar depletion was not affected in flagellar export (fliQ) and quorum-sensing (luxS) gene mutants. Passaged C. jejuni showed reduced invasiveness and increased serum sensitivity in vitro. C. jejuni alters its surface polysaccharides when cocultured with epithelial cells, suggesting the existence of a cross talk mechanism that modulates CPS expression during infection.

  4. Stem Cell Surface Marker Expression Defines Late Stages of Reprogramming to Pluripotency in Human Fibroblasts.

    Science.gov (United States)

    Pomeroy, Jordan E; Hough, Shelley R; Davidson, Kathryn C; Quaas, Alex M; Rees, Jordan A; Pera, Martin F

    2016-07-01

    Our current understanding of the induction of pluripotency by defined factors indicates that this process occurs in discrete stages characterized by specific alterations in the cellular transcriptome and epigenome. However, the final phase of the reprogramming process is incompletely understood. We sought to generate tools to characterize the transition to a fully reprogramed state. We used combinations of stem cell surface markers to isolate colonies emerging after transfection of human fibroblasts with reprogramming factors and then analyzed their expression of genes associated with pluripotency and early germ lineage specification. We found that expression of a subset of these genes, including the cell-cell adhesion molecule CDH3, characterized a late stage in the reprogramming process. Combined live-cell staining with the antibody GCTM-2 and anti-CDH3 during reprogramming identified colonies of cells that showed gene expression patterns very similar to those of embryonic stem cell or established induced pluripotent stem cell lines, and gave rise to stable induced pluripotent stem cell lines at high frequency. Our findings will facilitate studies of the final stages of reprogramming of human cells to pluripotency and will provide a simple means for prospective identification of fully reprogrammed cells. Reprogramming of differentiated cells back to an embryonic pluripotent state has wide ranging applications in understanding and treating human disease. However, how cells traverse the barriers on the journey to pluripotency still is not fully understood. This report describes tools to study the late stages of cellular reprogramming. The findings enable a more precise approach to dissecting the final phases of conversion to pluripotency, a process that is particularly poorly defined. The results of this study also provide a simple new method for the selection of fully reprogrammed cells, which could enhance the efficiency of derivation of cell lines for research

  5. Kettle holes formed by glacial outburst floods: identification when their surface expression has been removed?

    Science.gov (United States)

    Marren, Philip; Fay, Helen; Duller, Robert

    2014-05-01

    Kettle holes and obstacle marks formed by the transport, deposition and burial of ice-blocks during glacial outburst floods (jökulhlaups) are a common geomorphological feature on proglacial outwash plains. Indeed, they represent one of the few features which can unequivocally identify glacially-sourced flood deposits in the geomorphological and sedimentary record. Despite an abundance of work on the surface expression of jökulhlaup-generated ice-block structures, descriptions of the subsurface expression of these features in the sedimentary record are limited. There is currently no comprehensive model of the sedimentary characteristics of these features. This is a major gap in our knowledge, as the positive identification of ice-block features constitutes an unambiguous criterion for the identification of former jökulhlaup deposits in the Quaternary sedimentary record. We address this by describing several examples of ice-block impact in the sedimentary record from southern Iceland. Our work recognizes key criteria for the identification of ice-block impact in the sedimentary record, enabling them to be identified in sedimentary sections where their geomorphological expression has since been removed or buried. These key criterion combine: (1) structures formed by the interaction of water flow with the ice-block body during transportation and immobilization; (2) distinctive sedimentological features of surrounding deposits; and, (3) the post-burial mechanical disruption on the deposits. Formulating a suite of key criteria with which to positively identify the sedimentary impact of ice-blocks limits the possibility of misidentification in the sedimentary record, and provides a means of identifying previously unrecognized Quaternary catastrophic glacial floods.

  6. Role of Site-Specific N-Glycans Expressed on GluA2 in the Regulation of Cell Surface Expression of AMPA-Type Glutamate Receptors.

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    Yusuke Takeuchi

    Full Text Available The AMPA-type glutamate receptor (AMPAR, which is a tetrameric complex composed of four subunits (GluA1-4 with several combinations, mediates the majority of rapid excitatory synaptic transmissions in the nervous system. Cell surface expression levels of AMPAR modulate synaptic plasticity, which is considered one of the molecular bases for learning and memory formation. To date, a unique trisaccharide (HSO3-3GlcAβ1-3Galβ1-4GlcNAc, human natural killer-1 (HNK-1 carbohydrate, was found expressed specifically on N-linked glycans of GluA2 and regulated the cell surface expression of AMPAR and the spine maturation process. However, evidence that the HNK-1 epitope on N-glycans of GluA2 directly affects these phenomena is lacking. Moreover, it is thought that other N-glycans on GluA2 also have potential roles in the regulation of AMPAR functions. In the present study, using a series of mutants lacking potential N-glycosylation sites (N256, N370, N406, and N413 within GluA2, we demonstrated that the mutant lacking the N-glycan at N370 strongly suppressed the intracellular trafficking of GluA2 from the endoplasmic reticulum (ER in HEK293 cells. Cell surface expression of GluA1, which is a major subunit of AMPAR in neurons, was also suppressed by co-expression of the GluA2 N370S mutant. The N370S mutant and wild-type GluA2 were co-immunoprecipitated with GluA1, suggesting that N370S was properly associated with GluA1. Moreover, we found that N413 was the main potential site of the HNK-1 epitope that promoted the interaction of GluA2 with N-cadherin, resulting in enhanced cell surface expression of GluA2. The HNK-1 epitope on N-glycan at the N413 of GluA2 was also involved in the cell surface expression of GluA1. Thus, our data suggested that site-specific N-glycans on GluA2 regulate the intracellular trafficking and cell surface expression of AMPAR.

  7. Gene expression of human osteoblasts cells on chemically treated surfaces of Ti–6Al–4V–ELI

    Energy Technology Data Exchange (ETDEWEB)

    Oliveira, D.P., E-mail: dpedreira@ufscar.br [Department of Materials Engineering, Federal University of São Carlos, São Carlos (Brazil); Palmieri, A.; Carinci, F. [Department of D.M.C.C.C., Section of Maxillofacial and Plastic Surgery, University of Ferrara, Ferrara (Italy); Bolfarini, C. [Department of Materials Engineering, Federal University of São Carlos, São Carlos (Brazil)

    2015-06-01

    Surface modifications of titanium alloys are useful methods to enhance the biological stability of intraosseous implants and to promote a well succeeded osseointegration in the early stages of implantation. This work aims to investigate the influence of chemically modified surfaces of Ti–6Al–4V–ELI (extra-low interstitial) on the gene expression of human osteoblastic (HOb) cells. The surface treatments by acid etching or acid etching plus alkaline treatment were carried out to modify the topography, effective area, contact angle and chemical composition of the samples. The surface morphology was investigated using: scanning electron microscopy (SEM) and confocal laser-scanning microscope (CLSM). Roughness measurements and effective surface area were obtained using the CLSM. Surface composition was analysed by energy dispersive X-ray spectroscopy (EDX) and by X-Ray Diffraction (XRD). The expression levels of some bone related genes (ALPL, COL1A1, COL3A1, SPP1, RUNX2, and SPARC) were analysed using real-time Reverse Transcription Polymerase Chain Reaction (real-time RT-PCR). The results showed that all the chemical modifications studied in this work influenced the surface morphology, wettability, roughness, effective area and gene expression of human osteoblasts. Acid phosphoric combined to alkaline treatment presented a more accelerated gene expression after 7 days while the only phosphoric etching or chloride etching combined to alkaline treatment presented more effective responses after 15 days. - Highlights: • Chemical treatments were effective for surface modification of Ti–6Al–4V. • Alkaline and phosphoric treatments induced osteopontin up-regulation. • Topographic formation on surface can induce RUNX2 up regulation. • Acid etch plus alkaline treatment accelerated the expression of some bone related genes.

  8. Clostridium thermocellum Nitrilase Expression and Surface Display on Bacillus subtilis Spores.

    Science.gov (United States)

    Chen, Huayou; Zhang, Tianxi; Sun, Tengyun; Ni, Zhong; Le, Yilin; Tian, Rui; Chen, Zhi; Zhang, Chunxia

    2015-01-01

    Nitrilases are an important class of industrial enzymes. They require mild reaction conditions and are highly efficient and environmentally friendly, so they are used to catalyze the synthesis of carboxylic acid from nitrile, a process considered superior to conventional chemical syntheses. Nitrilases should be immobilized to overcome difficulties in recovery after the reaction and to stabilize the free enzyme. The nitrilase from Clostridium thermocellum was expressed, identified and displayed on the surface of Bacillus subtilis spores by using the spore coat protein G of B. subtilis as an anchoring motif. In a free state, the recombinant nitrilase catalyzed the conversion of 3-cyanopyridine to niacin and displayed maximum catalytic activity (8.22 units/mg protein) at 40 °C and pH 7.4. SDS-PAGE and Western blot were used to confirm nitrilase display. Compared with the free enzyme, the spore-immobilized nitrilase showed a higher tolerance for adverse environmental conditions. After the reaction, recombinant spores were recovered via centrifugation and reused 3 times to catalyze the conversion of 3-cyanopyridine with 75.3% nitrilase activity. This study demonstrates an effective means of nitrilase immobilization via spore surface display, which can be applied in biological processes or conversion. © 2015 S. Karger AG, Basel.

  9. Surface Expression of Precursor N-cadherin Promotes Tumor Cell Invasion

    Directory of Open Access Journals (Sweden)

    Deborah Maret

    2010-12-01

    Full Text Available The expression of N-cadherin (NCAD has been shown to correlate with increased tumor cell motility and metastasis. However, NCAD-mediated adhesion is a robust phenomenon and therefore seems to be inconsistent with the “release” from intercellular adhesion required for invasion. We show that in the most invasive melanoma and brain tumor cells, altered posttranslational processing results in abundant nonadhesive precursor N-cadherin (proNCAD at the cell surface, although total NCAD levels remain constant. We demonstrate that aberrantly processed proNCAD promotes cell migration and invasion in vitro. Furthermore, in human tumor specimens, we find high levels of proNCAD as well, supporting an overall conclusion that proNCAD and mature NCAD coexist on these tumor cell surfaces and that it is the ratio between these functionally antagonistic moieties that directly correlates with invasion potential. Our work provides insight into what may be a widespread mechanism for invasion and metastasis and challenges the current dogma of the functional roles played by classic cadherins in tumor progression.

  10. Analysis of Structures and Epitopes of Surface Antigen Glycoproteins Expressed in Bradyzoites of Toxoplasma gondii

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    Hua Cong

    2013-01-01

    Full Text Available Toxoplasma gondii is a protozoan parasite capable of infecting humans and animals. Surface antigen glycoproteins, SAG2C, -2D, -2X, and -2Y, are expressed on the surface of bradyzoites. These antigens have been shown to protect bradyzoites against immune responses during chronic infections. We studied structures of SAG2C, -2D, -2X, and -2Y proteins using bioinformatics methods. The protein sequence alignment was performed by T-Coffee method. Secondary structural and functional domains were predicted using software PSIPRED v3.0 and SMART software, and 3D models of proteins were constructed and compared using the I-TASSER server, VMD, and SWISS-spdbv. Our results showed that SAG2C, -2D, -2X, and -2Y are highly homologous proteins. They share the same conserved peptides and HLA-I restricted epitopes. The similarity in structure and domains indicated putative common functions that might stimulate similar immune response in hosts. The conserved peptides and HLA-restricted epitopes could provide important insights on vaccine study and the diagnosis of this disease.

  11. Turbulent characteristics of a semiarid atmospheric surface layer from cup anemometers – effects of soil tillage treatment (Northern Spain

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    S. Yahaya

    Full Text Available This paper deals with the characteristics of turbulent flow over two agricultural plots with various tillage treatments in a fallow, semiarid area (Central Aragon, Spain. The main dynamic characteristics of the Atmospheric Surface Layer (ASL measured over the experimental site (friction velocity, roughness length, etc., and energy budget, have been presented previously (Frangi and Richard, 2000. The current study is based on experimental measurements performed with cup anemometers located in the vicinity of the ground at 5 different levels (from 0.25 to 4 m and sampled at 1 Hz. It reveals that the horizontal wind variance, the Eulerian integral scales, the frequency range of turbulence and the turbulent kinetic energy dissipation rate are affected by the surface roughness. In the vicinity of the ground surface, the horizontal wind variance logarithmically increases with height, directly in relation to the friction velocity and the roughness length scale. It was found that the time integral scale (and subsequently the length integral scale increased with the surface roughness and decreased with the anemometer height. These variations imply some shifts in the meteorological spectral gap and some variations of the spectral peak length scale. The turbulent energy dissipation rate, affected by the soil roughness, shows a z-less stratification behaviour under stable conditions. In addition to the characterization of the studied ASL, this paper intends to show which turbulence characteristics, and under what conditions, are accessible through the cup anemometer.

    Key words. Meteorology and atmospheric dynamics (climatology, turbulence, instruments and techniques

  12. Turbulent characteristics of a semiarid atmospheric surface layer from cup anemometers – effects of soil tillage treatment (Northern Spain

    Directory of Open Access Journals (Sweden)

    S. Yahaya

    2003-10-01

    Full Text Available This paper deals with the characteristics of turbulent flow over two agricultural plots with various tillage treatments in a fallow, semiarid area (Central Aragon, Spain. The main dynamic characteristics of the Atmospheric Surface Layer (ASL measured over the experimental site (friction velocity, roughness length, etc., and energy budget, have been presented previously (Frangi and Richard, 2000. The current study is based on experimental measurements performed with cup anemometers located in the vicinity of the ground at 5 different levels (from 0.25 to 4 m and sampled at 1 Hz. It reveals that the horizontal wind variance, the Eulerian integral scales, the frequency range of turbulence and the turbulent kinetic energy dissipation rate are affected by the surface roughness. In the vicinity of the ground surface, the horizontal wind variance logarithmically increases with height, directly in relation to the friction velocity and the roughness length scale. It was found that the time integral scale (and subsequently the length integral scale increased with the surface roughness and decreased with the anemometer height. These variations imply some shifts in the meteorological spectral gap and some variations of the spectral peak length scale. The turbulent energy dissipation rate, affected by the soil roughness, shows a z-less stratification behaviour under stable conditions. In addition to the characterization of the studied ASL, this paper intends to show which turbulence characteristics, and under what conditions, are accessible through the cup anemometer.Key words. Meteorology and atmospheric dynamics (climatology, turbulence, instruments and techniques

  13. Cell cycle phase-specific surface expression of nerve growth factor receptors TrkA and p75(NTR).

    Science.gov (United States)

    Urdiales, J L; Becker, E; Andrieu, M; Thomas, A; Jullien, J; van Grunsven, L A; Menut, S; Evan, G I; Martín-Zanca, D; Rudkin, B B

    1998-09-01

    Expression of the nerve growth factor (NGF) receptors TrkA and p75(NTR) was found to vary at the surface of PC12 cells in a cell cycle phase-specific manner. This was evidenced by using flow cytometric and microscopic analysis of cell populations labeled with antibodies to the extracellular domains of both receptors. Differential expression of these receptors also was evidenced by biotinylation of surface proteins and Western analysis, using antibodies specific for the extracellular domains of TrkA and p75(NTR). TrkA is expressed most strongly at the cell surface in M and early G1 phases, whereas p75(NTR) is expressed mainly in late G1, S, and G2 phases. This expression reflects the molecular and cellular responses to NGF in specific phases of the cell cycle; in the G1 phase NGF elicits both the anti-mitogenic effect, i.e., inhibition of the G1 to S transition, and the differentiation response whereas a survival effect is provoked elsewhere in the cell cycle. A model is proposed relating these responses to the surface expression of the two receptors. These observations open the way for novel approaches to the investigation of the mechanism of NGF signal transduction.

  14. Differential Expression of Osteo-Modulatory Molecules in Periodontal Ligament Stem Cells in Response to Modified Titanium Surfaces

    Directory of Open Access Journals (Sweden)

    So Yeon Kim

    2014-01-01

    Full Text Available This study assessed differential gene expression of signaling molecules involved in osteogenic differentiation of periodontal ligament stem cells (PDLSCs subjected to different titanium (Ti surface types. PDLSCs were cultured on tissue culture polystyrene (TCPS, and four types of Ti discs (PT, SLA, hydrophilic PT (pmodPT, and hydrophilic SLA (modSLA with no osteoinductive factor and then osteogenic activity, including alkaline phosphatase (ALP activity, mRNA expression of runt-related gene 2, osterix, FOSB, FRA1, and protein levels of osteopontin and collagen type IA, were examined. The highest osteogenic activity appeared in PDLSCs cultured on SLA, compared with the TCPS and other Ti surfaces. The role of surface properties in affecting signaling molecules to modulate PDLSC behavior was determined by examining the regulation of Wnt pathways. mRNA expression of the canonical Wnt signaling molecules, Wnt3a and β-catenin, was higher on SLA and modSLA than on smooth surfaces, but gene expression of the calcium-dependent Wnt signaling molecules Wnt5a, calmodulin, and NFATc1 was increased significantly on PT and pmodPT. Moreover, integrin α2/β1, sonic hedgehog, and Notch signaling molecules were affected differently by each surface modification. In conclusion, surface roughness and hydrophilicity can affect differential Wnt pathways and signaling molecules, targeting the osteogenic differentiation of PDLSCs.

  15. Silkworm Apolipophorin Protein Inhibits Hemolysin Gene Expression of Staphylococcus aureus via Binding to Cell Surface Lipoteichoic Acids*

    Science.gov (United States)

    Omae, Yosuke; Hanada, Yuichi; Sekimizu, Kazuhisa; Kaito, Chikara

    2013-01-01

    We previously reported that a silkworm hemolymph protein, apolipophorin (ApoLp), binds to the cell surface of Staphylococcus aureus and inhibits expression of the saePQRS operon encoding a two-component system, SaeRS, and hemolysin genes. In this study, we investigated the inhibitory mechanism of ApoLp on S. aureus hemolysin gene expression. ApoLp bound to lipoteichoic acids (LTA), an S. aureus cell surface component. The addition of purified LTA to liquid medium abolished the inhibitory effect of ApoLp against S. aureus hemolysin production. In an S. aureus knockdown mutant of ltaS encoding LTA synthetase, the inhibitory effects of ApoLp on saeQ expression and hemolysin production were attenuated. Furthermore, the addition of anti-LTA monoclonal antibody to liquid medium decreased the expression of S. aureus saeQ and hemolysin genes. In S. aureus strains expressing SaeS mutant proteins with a shortened extracellular domain, ApoLp did not decrease saeQ expression. These findings suggest that ApoLp binds to LTA on the S. aureus cell surface and inhibits S. aureus hemolysin gene expression via a two-component regulatory system, SaeRS. PMID:23873929

  16. Silkworm apolipophorin protein inhibits hemolysin gene expression of Staphylococcus aureus via binding to cell surface lipoteichoic acids.

    Science.gov (United States)

    Omae, Yosuke; Hanada, Yuichi; Sekimizu, Kazuhisa; Kaito, Chikara

    2013-08-30

    We previously reported that a silkworm hemolymph protein, apolipophorin (ApoLp), binds to the cell surface of Staphylococcus aureus and inhibits expression of the saePQRS operon encoding a two-component system, SaeRS, and hemolysin genes. In this study, we investigated the inhibitory mechanism of ApoLp on S. aureus hemolysin gene expression. ApoLp bound to lipoteichoic acids (LTA), an S. aureus cell surface component. The addition of purified LTA to liquid medium abolished the inhibitory effect of ApoLp against S. aureus hemolysin production. In an S. aureus knockdown mutant of ltaS encoding LTA synthetase, the inhibitory effects of ApoLp on saeQ expression and hemolysin production were attenuated. Furthermore, the addition of anti-LTA monoclonal antibody to liquid medium decreased the expression of S. aureus saeQ and hemolysin genes. In S. aureus strains expressing SaeS mutant proteins with a shortened extracellular domain, ApoLp did not decrease saeQ expression. These findings suggest that ApoLp binds to LTA on the S. aureus cell surface and inhibits S. aureus hemolysin gene expression via a two-component regulatory system, SaeRS.

  17. Evaluation of Land Surface Models in Reproducing Satellite-Derived LAI over the High-Latitude Northern Hemisphere. Part I: Uncoupled DGVMs

    Directory of Open Access Journals (Sweden)

    Ning Zeng

    2013-10-01

    Full Text Available Leaf Area Index (LAI represents the total surface area of leaves above a unit area of ground and is a key variable in any vegetation model, as well as in climate models. New high resolution LAI satellite data is now available covering a period of several decades. This provides a unique opportunity to validate LAI estimates from multiple vegetation models. The objective of this paper is to compare new, satellite-derived LAI measurements with modeled output for the Northern Hemisphere. We compare monthly LAI output from eight land surface models from the TRENDY compendium with satellite data from an Artificial Neural Network (ANN from the latest version (third generation of GIMMS AVHRR NDVI data over the period 1986–2005. Our results show that all the models overestimate the mean LAI, particularly over the boreal forest. We also find that seven out of the eight models overestimate the length of the active vegetation-growing season, mostly due to a late dormancy as a result of a late summer phenology. Finally, we find that the models report a much larger positive trend in LAI over this period than the satellite observations suggest, which translates into a higher trend in the growing season length. These results highlight the need to incorporate a larger number of more accurate plant functional types in all models and, in particular, to improve the phenology of deciduous trees.

  18. Expression Level of the DREB2-Type Gene, Identified with Amplifluor SNP Markers, Correlates with Performance, and Tolerance to Dehydration in Bread Wheat Cultivars from Northern Kazakhstan

    Science.gov (United States)

    Shavrukov, Yuri; Zhumalin, Aibek; Serikbay, Dauren; Botayeva, Makpal; Otemisova, Ainur; Absattarova, Aiman; Sereda, Grigoriy; Sereda, Sergey; Shvidchenko, Vladimir; Turbekova, Arysgul; Jatayev, Satyvaldy; Lopato, Sergiy; Soole, Kathleen; Langridge, Peter

    2016-01-01

    A panel of 89 local commercial cultivars of bread wheat was tested in field trials in the dry conditions of Northern Kazakhstan. Two distinct groups of cultivars (six cultivars in each group), which had the highest and the lowest grain yield under drought were selected for further experiments. A dehydration test conducted on detached leaves indicated a strong association between rates of water loss in plants from the first group with highest grain yield production in the dry environment relative to the second group. Modern high-throughput Amplifluor Single Nucleotide Polymorphism (SNP) technology was applied to study allelic variations in a series of drought-responsive genes using 19 SNP markers. Genotyping of an SNP in the TaDREB5 (DREB2-type) gene using the Amplifluor SNP marker KATU48 revealed clear allele distribution across the entire panel of wheat accessions, and distinguished between the two groups of cultivars with high and low yield under drought. Significant differences in expression levels of TaDREB5 were revealed by qRT-PCR. Most wheat plants from the first group of cultivars with high grain yield showed slight up-regulation in the TaDREB5 transcript in dehydrated leaves. In contrast, expression of TaDREB5 in plants from the second group of cultivars with low grain yield was significantly down-regulated. It was found that SNPs did not alter the amino acid sequence of TaDREB5 protein. Thus, a possible explanation is that alternative splicing and up-stream regulation of TaDREB5 may be affected by SNP, but these hypotheses require additional analysis (and will be the focus of future studies). PMID:27917186

  19. Expression level of the DREB2-type gene, identified with Amplifluor SNP markers, correlates with performance and tolerance to dehydration in bread wheat cultivars from Northern Kazakhstan

    Directory of Open Access Journals (Sweden)

    Yuri Shavrukov

    2016-11-01

    Full Text Available A panel of 89 local commercial cultivars of bread wheat was tested in field trials in the dry conditions of Northern Kazakhstan. Two distinct groups of cultivars (six cultivars in each group, which had the highest and the lowest grain yield under drought were selected for further experiments. A dehydration test conducted on detached leaves indicated a strong association between rates of water loss in plants from the first group with highest grain yield production in the dry environment relative to the second group. Modern high-throughput Amplifluor SNP technology was applied to study allelic variations in a series of drought-responsive genes using 19 SNP markers. Genotyping of an SNP in the TaDREB5 (DREB2-type gene using the Amplifluor SNP marker KATU48 revealed clear allele distribution across the entire panel of wheat accessions, and distinguished between the two groups of cultivars with high and low yield under drought. Significant differences in expression levels of TaDREB5 were revealed by qRT-PCR. Most wheat plants from the first group of cultivars with high grain yield showed strong up-regulation of TaDREB5 transcript in dehydrated leaves. In contrast, expression of TaDREB5 in plants from the second group of cultivars with low grain yield was significantly down-regulated. It was found that SNPs did not alter the amino acid sequence of TaDREB5 protein. Thus, a possible explanation is that alternative splicing and up-stream regulation of TaDREB5 may be affected by SNP, but these hypotheses require additional analysis (and will be the focus of future studies.

  20. Surface glycoprotein PSA (GP46) expression during short- and long-term culture of Leishmania chagasi.

    Science.gov (United States)

    Beetham, Jeffrey K; Donelson, John E; Dahlin, Rebecca R

    2003-10-01

    The mRNAs encoding promastigote surface antigen (PSA) of Leishmania chagasi have previously been shown to increase about 30-fold as in vitro cultured parasites progress from logarithmic to stationary phase, growth phases that are, respectively associated with parasites having low and high infectivity to mammals. Experiments reported here establish by western blot analysis that PSA proteins of 44 and 66 kDa also increase about 30-fold as parasite cultures reach stationary phase. Serial passage of parasite cultures resulted in a progressive reduction in PSA protein and RNA abundance to levels less than 3% that of cultures newly-initiated with parasites derived from a parasitized rodent. Loss of PSA mRNA abundance in serially passaged cells was not due to reduced PSA gene transcription rates, as determined by nuclear run-on assays. Neither was the loss associated with a marked decrease in PSA mRNA stability. Analysis of PSA RNA stability in the presence of actinomycin D, an inhibitor of transcription elongation, failed to detect a difference in fully processed cytosolic PSA mRNA stability regardless of the number of times a culture was passaged or the growth phase of the culture. Based on the lack of detectable difference in (cytosolic) mature PSA mRNA stability during promastigote development, the data indirectly suggest that the regulated expression of PSA in cells from low-passage cultures and the loss of PSA expression in high-passage cultures may be mediated by nuclear events that occur after transcription of the PSA genes and before arrival of the mature mRNAs in the cytoplasm.

  1. Variations in pCO2 during summer in the surface water of an unproductive lake in northern Sweden

    International Nuclear Information System (INIS)

    Jonsson, A.; Aaberg, J.; Jansson, M.

    2007-01-01

    Unproductive lakes are generally supersaturated with carbon dioxide (CO 2 ) and emit CO 2 to the atmosphere continuously during ice-free periods. However, temporal variation of the partial pressure of CO 2 (pCO 2 ) and thus of CO 2 evasion to atmosphere is poorly documented. We therefore carried out temporally high-resolution (every 6 h) measurements of the pCO 2 using an automated logger system in the surface water of a subarctic, unproductive, lake in the birch forest belt. The study period was June-September 2004. We found that the pCO 2 showed large seasonal variation, but low daily variation. The seasonal variation was likely mainly caused by variations in input and mineralization of allochthonous organic matter. Stratification depth probably also influenced pCO 2 of the surface water by controlling the volume in which mineralization of dissolved organic carbon (DOC) occurred. In lakes, with large variations in pCO 2 , as in our study lake a high (weekly) sampling intensity is recommended for obtaining accurate estimates of the evasion of CO 2

  2. The impact of land initialization on seasonal forecasts of surface air temperature: role of snow data assimilation in the Northern Hemisphere

    Science.gov (United States)

    Lin, P.; Wei, J.; Zhang, Y.; Yang, Z. L.

    2015-12-01

    Land initializations (i.e., snow, soil moisture, leaf area index) have been recognized as important sources of seasonal climate predictability besides ocean and atmosphere initializations. However, studies focusing on assessing how land data assimilation (DA) contributes to seasonal forecast skills are still lacking due to the limited number of large-scale land DA studies. In this study, taking advantage of the snow outputs from a multivariate global land DA system (i.e., DART/CLM), we systematically investigated the role of large-scale snow DA in influencing seasonal forecasts of surface air temperature. Three suites of ensemble seasonal forecast experiments were performed using the Community Earth System Model (CESM v1.2.1), in which three different snow initialization datasets were used. They are (1) CLM4 simulation without DA, (2) CLM4 simulation with MODIS snow cover DA, and (3) CLM4 simulation with joint GRACE and MODIS snow DA. Each suite of the experiment starts from multiple initialization dates of eight years from 2003 to 2010 and has three-month lead times. All experiments used the same atmosphere initializations from ERA-Interim (perturbed to get 8 ensembles) and the same prescribed SSTs. Our results show that snow DA plays an important role in surface air temperature predictions in regions such as Europe, western Canada, northern Alaska, Mongolia Plateau, Tibetan Plateau, and the Rocky Mountains. The analyses also account for multiple lead times as snow can influence the atmosphere through immediate snow-albedo effect and through delayed snow hydrological effect after snow melts and wets the soil. This is a first study to quantify the impacts of snow initializations on seasonal forecasts of surface air temperature with an emphasis on large-scale snow DA. The insights are helpful to both land DA studies as well as research on seasonal climate forecasts.

  3. Contribution of GIS to evaluate surface water pollution by heavy metals: Case of Ichkeul Lake (Northern Tunisia)

    Science.gov (United States)

    Yazidi, Amira; Saidi, Salwa; Ben Mbarek, Nabiha; Darragi, Fadila

    2017-10-01

    The concentrations of nutrients and heavy elements in the surface water of the lake Ichkeul, main wadis which feed directly and thermal springs that flow into the lake, are measured to evaluate these chemical elements. There are used to highlight the interactions between these different aquatic compartments of Ichkeul. All metal concentrations in lake water, except Cu, were lower than the maximum permitted concentration for the protection of aquatic life. The results show that the highest concentrations are located in the eastern and south-eastern part of the lake where the polluted water comes from the lagoon of Bizerte through the wadi Tinja as well as from the city of Mateur through the wadi Joumine. The pollution indices and especially the heavy metal evaluation index (HEI) show high pollution specially located at the mouths of wadis and an increase of heavy metal concentrations, as a result of uncontrolled releases of domestic and industrial wastewater.

  4. Surface engineered magnetic nanoparticles for specific immunotargeting of cadherin expressing cells

    International Nuclear Information System (INIS)

    Moros, Maria; Puertas, Sara; Saez, Berta; Grazú, Valeria; Delhaes, Flavien; Feracci, Helene; De la Fuente, Jesús M

    2016-01-01

    In spite of historic advances in cancer biology and recent development of sophisticated chemotherapeutics, the outlook for patients with advanced cancer is still grim. In this sense nanoparticles (NPs), through their unique physical properties, enable the development of new approaches for cancer diagnosis and treatment. Thus far the most used active targeting scheme involves NPs functionalization with antibodies specific to molecules overexpressed on cancer cell’s surface. Therefore, such active targeting relies on differences in NPs uptake kinetics rates between tumor and healthy cells. Many cancers of epithelial origin are associated with the inappropriate expression of non-epithelial cadherins (e.g. N-, P-, -11) with concomitant loss of E-cadherin. Such phenomenon named cadherin switching favors tumor development and metastasis via interactions of tumor cells with stromal components. That is why we optimized the oriented functionalization of fluorescently labelled magnetic NPs with a novel antibody specific for the extracellular domain of cadherin-11. The obtained Ab-NPs exhibited high specificity when incubated with two cell lines used as models of tumor and healthy cells. Thus, cadherin switching offers a great opportunity for the development of active targeting strategies aimed to improve the early detection and treatment of cancer. (paper)

  5. Impaired cell surface expression of HLA-B antigens on mesenchymal stem cells and muscle cell progenitors

    DEFF Research Database (Denmark)

    Isa, Adiba; Nehlin, Jan; Sabir, Hardee Jawad

    2010-01-01

    HLA class-I expression is weak in embryonic stem cells but increases rapidly during lineage progression. It is unknown whether all three classical HLA class-I antigens follow the same developmental program. In the present study, we investigated allele-specific expression of HLA-A, -B, and -C...... at the mRNA and protein levels on human mesenchymal stem cells from bone marrow and adipose tissue as well as striated muscle satellite cells and lymphocytes. Using multicolour flow cytometry, we found high cell surface expression of HLA-A on all stem cells and PBMC examined. Surprisingly, HLA-B was either...... undetectable or very weakly expressed on all stem cells protecting them from complement-dependent cytotoxicity (CDC) using relevant human anti-B and anti-Cw sera. IFNgamma stimulation for 48-72 h was required to induce full HLA-B protein expression. Quantitative real-time RT-PCR showed that IFNgamma induced...

  6. Transcript expression analysis of putative Trypanosoma brucei GPI-anchored surface proteins during development in the tsetse and mammalian hosts.

    Directory of Open Access Journals (Sweden)

    Amy F Savage

    Full Text Available Human African Trypanosomiasis is a devastating disease caused by the parasite Trypanosoma brucei. Trypanosomes live extracellularly in both the tsetse fly and the mammal. Trypanosome surface proteins can directly interact with the host environment, allowing parasites to effectively establish and maintain infections. Glycosylphosphatidylinositol (GPI anchoring is a common posttranslational modification associated with eukaryotic surface proteins. In T. brucei, three GPI-anchored major surface proteins have been identified: variant surface glycoproteins (VSGs, procyclic acidic repetitive protein (PARP or procyclins, and brucei alanine rich proteins (BARP. The objective of this study was to select genes encoding predicted GPI-anchored proteins with unknown function(s from the T. brucei genome and characterize the expression profile of a subset during cyclical development in the tsetse and mammalian hosts. An initial in silico screen of putative T. brucei proteins by Big PI algorithm identified 163 predicted GPI-anchored proteins, 106 of which had no known functions. Application of a second GPI-anchor prediction algorithm (FragAnchor, signal peptide and trans-membrane domain prediction software resulted in the identification of 25 putative hypothetical proteins. Eighty-one gene products with hypothetical functions were analyzed for stage-regulated expression using semi-quantitative RT-PCR. The expression of most of these genes were found to be upregulated in trypanosomes infecting tsetse salivary gland and proventriculus tissues, and 38% were specifically expressed only by parasites infecting salivary gland tissues. Transcripts for all of the genes specifically expressed in salivary glands were also detected in mammalian infective metacyclic trypomastigotes, suggesting a possible role for these putative proteins in invasion and/or establishment processes in the mammalian host. These results represent the first large-scale report of the differential

  7. Rifting mechanisms constrained by InSAR, seismicity, GPS, and surface rupture from the Karonga earthquake sequence in northern Lake Malawi (Nyasa)

    Science.gov (United States)

    Zheng, W.; Pritchard, M. E.; Henderson, S. T.; Gaherty, J. B.; Shillington, D. J.; Oliva, S. J.; Ebinger, C.; Nooner, S. L.; Elliott, J.; Saria, E.; Ntambila, D.; Chindandali, P. R. N.

    2017-12-01

    The Malawi rift is part of the archetypal East African rift where early-stage crustal extension is dominated by faulting. In the Karonga region of northern Malawi, a sequence of earthquakes in late 2009, with 15 teleseismically detected (Mw 4.5-6.0) over 13 days, provides a uniqueopportunity to evaluate faulting processes controlling present-day extension in an early-stage rift. We describe observations of this sequence including hundreds of aftershocks located by a temporary seismic array installed in 2010, ground deformation from satellite interferograms, and surface rupture from field surveys published by others. We use all of these data to model fault geometry and slip. The aftershocks from January-May 2010 suggest the involvement of multiple faults, and we test the extent that this can be resolved by the InSAR data. The InSAR and surface rupture both suggest that the major slip occurred at shallow depth (Study of Extension and maGmatism in Malawi aNd Tanzania) project, which includes the Karonga region and spans 2013-2015. We find 3 cm of potential ground movement at the location of the earthquake as determined by the SEGMeNT seismic array from Sentinel-1. Geodetic fault slip is consistent with the focal mechanism and depth determined by the local array. The location is at the northern end of the 2009-2010 aftershock zone, and aftershocks suggest some linkage with faults that slipped in 2009. InSAR observations do not provide any evidence for large aseismic slip or fluid movements during or after the 2014 sequence, which had <200 aftershocks above the network threshold. For example, we do not observe any deformation at Rungwe volcano above the 2 cm/yr detection threshold with InSAR time series from ALOS (2007-2010) or Sentinel-1 (10/2014 - 04/2017). The time series from SEGMeNT and other continuous GPS stations do not show transients related to the earthquakes, but are not optimally located in space or time.

  8. Expression and Purification of Glycosyltransferases in Pichia Pastoris: Towards Improving the Migration of Stem Cells by Enhancing Surface Expression of Sialyl Lewis X

    KAUST Repository

    Al-Amoodi, Asma S.

    2017-05-01

    Recruitment of circulating cells towards target sites is primarily dependent on E-selectin receptor/ligand adhesive interactions. Glycosyltransferase (GTs) are involved in the creation of E-selectin ligands. A sialofucosylated terminal tetrasaccharide like glycan structure known as sialyl Lewis x (sLex), is the most recognized ligand by selectins. This structure is found on the surface of cancer cells and leukocytes but is often absent on the surface of many adult stem cell populations. In order to synthesize sLex, GTs must be endogenously expressed and remain active within the cells. Generally, these stem cells express terminal sialylated lactosamine structures on their glycoproteins which require the addition of alpha-(1,3)-fucose to be converted into an E-selectin ligand. There are a number of fucosyltransferases (FUTs) that are able to modify terminal lactosamine structures to create sLex such as FUT6. In this work we focused on expressing and purifying active recombinant FUTs as a tool to help create sLex structures on the surface of adult stem cells in order to enhance their migration.

  9. The Synergic Characteristics of Surface Water Pollution and Sediment Pollution with Heavy Metals in the Haihe River Basin, Northern China

    Directory of Open Access Journals (Sweden)

    Peiru Kong

    2018-01-01

    Full Text Available Aquatic environmental deterioration is becoming a serious problem due to rapid urbanization and economic development, particularly in developing countries. As two important components of the aquatic environment, water quality and sediment pollution are widely considered to be concerns; however, they are considered separately in most cases. The relationship between water quality and sediment pollution with heavy metals has been little addressed. In this study, the Haihe River Basin (HRB, one of the most polluted areas in China, was used as a case study, and the eutrophication index (EI and the potential ecological risk index (RI were employed to evaluate water quality and sediment pollution of heavy metals, respectively. The results showed that generally in the HRB, the water quality was poor, while the risk of heavy metal pollution was relatively low. Surface water quality was mainly influenced by sewage discharges from human daily life, and heavy metal pollution was affected by industry structure, in that the areas with resource/energy consumption industries and high-pollution industries often have high risks of heavy metal pollution Synergic pollution from water eutrophication and sediment pollution with heavy metals was found, especially in the central areas of the HRB, and it was largely dependent on the type of human activities. In the places with intensive human activities, such as secondary industry, eutrophication occurred simultaneously with heavy metal pollution, other than in less human-affected areas. These findings are useful for planning aquatic environment protections and river ecosystem management.

  10. Prostaglandin D2 and Interleukin-5 Reduce Crth2 Surface Expression on Human Eosinophils

    Directory of Open Access Journals (Sweden)

    Kazuyuki Hamada

    2004-01-01

    Conclusions: These results suggest that PGD2 and IL-5 regulate CRTH2 expression on eosinophils through CRTH2 internalization. The decreased expression of CRTH2 on tissue eosinophils may make these cells remain at the site of allergic inflammation.

  11. Benzene and toluene in the surface air of northern Eurasia from TROICA-12 campaign along the Trans-Siberian Railway

    Science.gov (United States)

    Skorokhod, Andrey I.; Berezina, Elena V.; Moiseenko, Konstantin B.; Elansky, Nikolay F.; Belikov, Igor B.

    2017-05-01

    Volatile organic compounds (VOCs) were measured by proton transfer reaction mass spectrometry (PTR-MS) on a mobile laboratory in a transcontinental TROICA-12 (21 July-4 August 2008) campaign along the Trans-Siberian Railway from Moscow to Vladivostok. Surface concentrations of benzene (C6H6) and toluene (C7H8) along with non-methane hydrocarbons (NMHCs), CO, O3, SO2, NO, NO2 and meteorology are analyzed in this study to identify the main sources of benzene and toluene along the Trans-Siberian Railway. The most measurements in the TROICA-12 campaign were conducted under low-wind/stagnant conditions in moderately ( ˜ 78 % of measurements) to weakly polluted ( ˜ 20 % of measurements) air directly affected by regional anthropogenic sources adjacent to the railway. Only 2 % of measurements were identified as characteristic of highly polluted urban atmosphere. Maximum values of benzene and toluene during the campaign reached 36.5 and 45.6 ppb, respectively, which is significantly less than their short-term exposure limits (94 and 159 ppb for benzene and toluene, respectively). About 90 % of benzene and 65 % of toluene content is attributed to motor vehicle transport and 10 and 20 %, respectively, provided by the other local- and regional-scale sources. The highest average concentrations of benzene and toluene are measured in the industrial regions of the European Russia (up to 0.3 and 0.4 ppb for benzene and toluene, respectively) and south Siberia (up to 0.2 and 0.4 ppb for benzene and toluene, respectively). Total contribution of benzene and toluene to photochemical ozone production along the Trans-Siberian Railway is about 16 % compared to the most abundant organic VOC - isoprene. This contribution, however, is found to be substantially higher (up to 60-70 %) in urbanized areas along the railway, suggesting an important role of anthropogenic pollutant sources in regional ozone photochemistry and air quality.

  12. ِIncreased expression of T- cell- surface CXCR4 in asthmatic children.

    African Journals Online (AJOL)

    Ehab

    express CXCR 3 and CCR 5, whereas Th-2 subset expresses CCR3, CCR4, CCR8 and CXCR4 receptors10. Of these, Th-2 cell-expressed chemokine receptors, CXCR4, and its ligand SDF-. 1, have been shown to be the most relevant for Th-. 2-type allergic airway responses 11. The results presented demonstrate that.

  13. Divalent cations and the protein surface co-ordinate the intensity of human platelet adhesion and P-selectin surface expression.

    Science.gov (United States)

    Whiss, P A; Andersson, R G G

    2002-07-01

    At sites of blood vessel injury, platelets adhere to exposed vessel components, such as collagen, or immobilized fibrinogen derived from plasma or activated platelets. The divalent cations Mg(2+) and Ca(2+) are essential for platelet adhesion and activation, but Mg(2+) can also inhibit platelet activation. The present study evaluates, by an enzymatic method, the effects of various divalent cations on the adhesion of isolated human platelets to collagen, fibrinogen, albumin or plastic in vitro. By enzyme-linked immunosorbent assay, platelet surface expression of P-selectin was measured to estimate the state of activation on adherence. Mg(2+) increased platelet adhesion exclusively to collagen and fibrinogen at physiologically relevant concentrations. At higher concentrations, the adhesion declined. Ca(2+) induced a weak adhesion only to fibrinogen at physiological doses and a peak of increased adhesion to all protein-coated surfaces at 10 mmol/l. Mn(2+) elicited dose-dependent adhesion only to collagen and fibrinogen. Zn(2+), Ni(2+) and Cu(2+) increased the adhesion of platelets independently of the surface. Ca(2+) dose-dependently inhibited adhesion elicited by Mg(2+) to collagen and fibrinogen. No other combination of divalent cations elicited such an effect. Mg(2+)-dependent platelet adhesion to collagen and Ca(2+)-dependent adhesion to fibrinogen increased P-selectin expression. Thus, the present study shows that the outcome of the platelet adhesion depends on the surface and the access of divalent cations, which co-ordinate the intensity of platelet adhesion and P-selectin surface expression.

  14. Comparison of surface energy budgets and feedbacks to microclimate among different land use types in an agro-pastoral ecotone of northern China.

    Science.gov (United States)

    Zhao, Wei; Hu, Zhongmin; Li, Shenggong; Guo, Qun; Liu, Zhengjia; Zhang, Leiming

    2017-12-01

    The biophysical effect of land use conversion plays a significant role in regulating climate change. Owing to albedo and evapotranspiration (ET) change, the effect of energy budget difference on land surface temperature (LST) is important but unclear among contrasting land use types, especially in temperate semi-arid regions. Based on moderate-resolution imaging spectroradiometer (MODIS) data, we compared the differences in albedo, ET, and LST between cropland and grassland (CR-GR), and between planted forest and grassland (PF-GR) in the Horqin Sandy Land of Inner Mongolia, an agro-pastoral ecotone of northern China. Our main objective was to explore the magnitude and direction of albedo and ET change during the growing season and, subsequently, to estimate the biophysical effects on LST as a result of land use and land cover change. Our results indicate no significant difference in mean monthly albedo for CR-GR and PF-GR. Cropland lost more water through ET and significantly decreased daytime LST compared with grassland from July to September, but no significant differences in ET and LST were observed for PF-GR in any month. The biophysical climate effects were more pronounced for CR-GR compared with PF-GR. The response of LST to the changes in energy budget confirmed that ET was the critical driving factor relative to albedo. Compared with grassland, cropland and planted forest tended to cool the land surface by 5.15°C and 1.51°C during the growing season, respectively, because of the biophysical effects. Our findings suggest the significance of local-scale biophysical effect on climate variation after land use conversion in semi-arid regions. Copyright © 2017 Elsevier B.V. All rights reserved.

  15. How to build the Eiger: Surface expression of litho-tectonic preconditioning

    Science.gov (United States)

    Mair, David; Lechmann, Alessandro; Schlunegger, Fritz

    2017-04-01

    The north face of the Eiger has exerted a strong attraction on alpinists, but also on geologists during the past decades, mainly because of its triangular, nearly vertical shape. We build on this tradition and investigate the relationship between the shape of this mountain and its underlying lithology, and its history of folding and thrusting. To this extent, we constructed a geometric 3D geological model of the Eiger-Moench-Jungfrau mountain chain in the central Swiss Alps. We proceeded through compilations of geological maps that we combined with new mapping in the field and collection of structural data such as the orientation of lineaments and faults. The model itself was constructed by interpolation of interfaces between geological formations, thrust- and fold-geometries between several NW-SE running, balanced, cross-sections. In addition, new geological data from the Jungfraubahn railway tunnel was used to verify surface data and improve the resulting model in the depth. The analyzed units of the Hercynian crystalline basement of the Aar massif and the Mesozoic cover rocks of the Helvetics form a foliated and thrusted stack. Multiple ductile structure sets bear witness of Alpine deformation and are dominant amid the mark of later brittle deformation across the whole mountain. There are two major outcomes of this analysis. First, the thrust contact between two stacks, which comprise a foliated basement and cover rocks, are responsible for the shape and overall architecture of the Eiger and its famous north face. Second, the high-resolution 3D structural model paired with petrological data shows that second-order, horizontally aligned morphological steps in the north face are related to the foliation within the bedrock. We suspect the inherited fabric significantly modified the susceptibility to erosion mechanisms which in turn further amplified the morphological differences (expressed in e.g. terrain roughness or slope).

  16. Surface EMG-based Sketching Recognition Using Two Analysis Windows and Gene Expression Programming

    Science.gov (United States)

    Yang, Zhongliang; Chen, Yumiao

    2016-01-01

    Sketching is one of the most important processes in the conceptual stage of design. Previous studies have relied largely on the analyses of sketching process and outcomes; whereas surface electromyographic (sEMG) signals associated with sketching have received little attention. In this study, we propose a method in which 11 basic one-stroke sketching shapes are identified from the sEMG signals generated by the forearm and upper arm muscles from 4 subjects. Time domain features such as integrated electromyography, root mean square and mean absolute value were extracted with analysis windows of two length conditions for pattern recognition. After reducing data dimensionality using principal component analysis, the shapes were classified using Gene Expression Programming (GEP). The performance of the GEP classifier was compared to the Back Propagation neural network (BPNN) and the Elman neural network (ENN). Feature extraction with the short analysis window (250 ms with a 250 ms increment) improved the recognition rate by around 6.4% averagely compared with the long analysis window (2500 ms with a 2500 ms increment). The average recognition rate for the eleven basic one-stroke sketching patterns achieved by the GEP classifier was 96.26% in the training set and 95.62% in the test set, which was superior to the performance of the BPNN and ENN classifiers. The results show that the GEP classifier is able to perform well with either length of the analysis window. Thus, the proposed GEP model show promise for recognizing sketching based on sEMG signals. PMID:27790083

  17. Enhanced expression of rabies virus surface G-protein in Escherichia coli using SUMO fusion.

    Science.gov (United States)

    Singh, Ankit; Yadav, Dinesh; Rai, Krishan Mohan; Srivastava, Meenal; Verma, Praveen C; Singh, Pradhyumna K; Tuli, Rakesh

    2012-01-01

    Fusion systems are known to increase the expression of difficult to express recombinant proteins in soluble form to facilitate their purification. Rabies glycoprotein was also tough to express at sufficient level in soluble form in both E. coli and plant. The present work was aimed to over-express and purify this membrane protein from soluble extract of E. coli. Fusion of Small Ubiqutin like Modifier (SUMO) with rabies glycoprotein increased ~1.5 fold higher expression and ~3.0 fold solubility in comparison to non-fused in E. coli. The SUMO fusion also simplified the purification process. Previously engineered rabies glycoprotein gene in tobacco plants provides complete protection to mice, but the expression was very low for purification. Our finding demonstrated that the SUMO-fusion was useful for enhancing expression and solubility of the membrane protein and again proves to be a good alternative technology for applications in biomedical and pharmaceutical research.

  18. Elucidating the Kinetics of Expression and Immune Cell Infiltration Resulting from Plasmid Gene Delivery Enhanced by Surface Dermal Electroporation

    Directory of Open Access Journals (Sweden)

    Kate E. Broderick

    2013-08-01

    Full Text Available The skin is an attractive tissue for vaccination in a clinical setting due to the accessibility of the target, the ease of monitoring and most importantly the immune competent nature of the dermal tissue. While skin electroporation offers an exciting and novel future methodology for the delivery of DNA vaccines in the clinic, little is known about the actual mechanism of the approach and the elucidation of the resulting immune responses. To further understand the mechanism of this platform, the expression kinetics and localization of a reporter plasmid delivered via a surface dermal electroporation (SEP device as well as the effect that this treatment would have on the resident immune cells in that tissue was investigated. Initially a time course (day 0 to day 21 of enhanced gene delivery with electroporation (EP was performed to observe the localization of green fluorescent protein (GFP expression and the kinetics of its appearance as well as clearance. Using gross imaging, GFP expression was not detected on the surface of the skin until 8 h post treatment. However, histological analysis by fluorescent microscopy revealed GFP positive cells as early as 1 h after plasmid delivery and electroporation. Peak GFP expression was observed at 24 h and the expression was maintained in skin for up to seven days. Using an antibody specific for a keratinocyte cell surface marker, reporter gene positive keratinocytes in the epidermis were identified. H&E staining of treated skin sections demonstrated an influx of monocytes and granulocytes at the EP site starting at 4 h and persisting up to day 14 post treatment. Immunological staining revealed a significant migration of lymphocytic cells to the EP site, congregating around cells expressing the delivered antigen. In conclusion, this study provides insights into the expression kinetics following EP enhanced DNA delivery targeting the dermal space. These findings may have implications in the future to design

  19. Low-latitude expressions of high-latitude forcing during Heinrich Stadial 1 and the Younger Dryas in northern South America

    Science.gov (United States)

    Bahr, André; Hoffmann, Julia; Schönfeld, Joachim; Schmidt, Matthew W.; Nürnberg, Dirk; Batenburg, Sietske J.; Voigt, Silke

    2018-01-01

    Changes in Atlantic Meridional Overturning Circulation (AMOC) strength exert a major influence on global atmospheric circulation patterns. However, the pacing and mechanisms of low-latitude responses to high-latitude forcing are insufficiently constrained so far. To elucidate the interaction of atmospheric and oceanic forcing in tropical South America during periods of major AMOC reductions (Heinrich Stadial 1 and the Younger Dryas) we generated a high-resolution foraminiferal multi-proxy record from off the Orinoco River based on Ba/Ca and Mg/Ca ratios, as well as stable isotope measurements. The data clearly indicate a three-phased structure of HS1 based on the reconfiguration of ocean currents in the tropical Atlantic Ocean. The initial phase (HS1a) is characterized by a diminished North Brazil Current, a southward displacement of the ITCZ, and moist conditions dominating northeastern Brazil. During subsequent HS1b, the NBC was even more diminished or yet reversed and the ITCZ shifted to its southernmost position. Hence, dryer conditions prevailed in northern South America, while eastern Brazil experienced maximally wet conditions. During the final stage, HS1c, conditions are similar to HS1a. The YD represents a smaller amplitude version of HS1 with a southward-shifted ITCZ. Our findings imply that the low-latitude continental climate response to high-latitude forcing is mediated by reconfigurations of surface ocean currents in low latitudes. Our new records demonstrate the extreme sensitivity of the terrestrial realm in tropical South America to abrupt perturbations in oceanic circulation during periods of unstable climate conditions.

  20. Salmonella flagellin acted as an effective fusion partner for expression of Plasmodium falciparum surface protein 25 in Escherichia coli.

    Science.gov (United States)

    Qian, Feng; Li, Mengmeng; Chen, Yong; Jiang, Lin; Xu, Huji

    2016-09-01

    Plasmodium falciparum surface protein 25 (Pfs25) is a hard-to-express and hard-to-solubilize protein in Escherichia coli. To overcome this problem, the phase 1 flagellin of Salmonella enterica serovar Typhimurium (FliC) was used as a fusion partner for Pfs25. The fusion expression of Pfs25 with FliC greatly enhanced the expression level and solubility of Pfs25 in E. coli BL21(DE3). The Ni-purified fusion protein of FliC-Pfs25 was recognized by two anti-Pfs25 monoclonal antibodies. By comparison, it was shown that the Pfs25 within FliC-Pfs25 contained epitopes similar or identical to those on Pichia pastoris-produced Pfs25. The data obtained from this study demonstrated that the fusion with Salmonella flagellin greatly improved the expression of Pfs25 in E. coli.

  1. Comparative analysis of the expression of surface markers on fibroblasts and fibroblast-like cells isolated from different human tissues.

    Science.gov (United States)

    Lupatov, A Yu; Vdovin, A S; Vakhrushev, I V; Poltavtseva, R A; Yarygin, K N

    2015-02-01

    Expression of 20 surface markers was analyzed in cultures of mesenchymal stromal cells of the umbilical cord, fibroblasts from adult and fetal human skin, and fibroblast-like cells of fetal liver was analyzed by fl ow cytometry. The studied cultures did not express hemopoietic cells markers, but were positive for CD73, CD90, and CD105 markers recommended by the International Society of Cell Therapy for the identification of the multipotent mesenchymal stromal cells. Fetal liver fibroblast-like cells were positive for CD54; this marker was absent in skin fibroblast cultures, but was expressed by umbilical cord mesenchymal stromal cells. Further study of these cells revealed a minor subpopulation of cells co-expressing CD24 and CD90 or CD24 and CD54. We hypothesized that these cells probably participate in epithelial mesenchymal transition.

  2. Expression

    Directory of Open Access Journals (Sweden)

    Wang-Xia Wang

    2014-02-01

    Full Text Available The miR-15/107 family comprises a group of 10 paralogous microRNAs (miRNAs, sharing a 5′ AGCAGC sequence. These miRNAs have overlapping targets. In order to characterize the expression of miR-15/107 family miRNAs, we employed customized TaqMan Low-Density micro-fluid PCR-array to investigate the expression of miR-15/107 family members, and other selected miRNAs, in 11 human tissues obtained at autopsy including the cerebral cortex, frontal cortex, primary visual cortex, thalamus, heart, lung, liver, kidney, spleen, stomach and skeletal muscle. miR-103, miR-195 and miR-497 were expressed at similar levels across various tissues, whereas miR-107 is enriched in brain samples. We also examined the expression patterns of evolutionarily conserved miR-15/107 miRNAs in three distinct primary rat brain cell preparations (enriched for cortical neurons, astrocytes and microglia, respectively. In primary cultures of rat brain cells, several members of the miR-15/107 family are enriched in neurons compared to other cell types in the central nervous system (CNS. In addition to mature miRNAs, we also examined the expression of precursors (pri-miRNAs. Our data suggested a generally poor correlation between the expression of mature miRNAs and their precursors. In summary, we provide a detailed study of the tissue and cell type-specific expression profile of this highly expressed and phylogenetically conserved family of miRNA genes.

  3. Ionizing radiation modulates the surface expression of human leukocyte antigen-G in a human melanoma cell line

    Energy Technology Data Exchange (ETDEWEB)

    Michelin, S.; Gallegos, C.E.; Dubner, D. [Radiopathology Laboratory, Nuclear Regulatory Authority, Buenos Aires (Argentina); Favier, B.; Carosella, E.D. [CEA, I2BM, Hopital Saint-Louis, IUH, Service de Recherches en Hemato-Immunologie, Paris (France)

    2009-07-01

    Human leukocyte antigen G (HLA-G) is a nonclassical HLA class I molecule involved in fetus protection from the maternal immune system, transplant tolerance, and viral and tumoral immune escape. Tumor-specific HLA-G expression has been described for a wide variety of malignancies, including melanomas. The aim of this study was to evaluate whether ionizing radiation (IR) could modulate the surface expression of HLA-G1 in a human melanoma cell line that expresses endogenously membrane-bound HLA-G1. For this purpose, cells were exposed to increasing doses of {gamma}-irradiation (0-20 Gy) and HLA-G1 levels at the plasma membrane were analyzed at different times postirradiation by flow cytometry. HLA-G total expression and the presence of the soluble form of HLA-G1 (sHLA-G1) in the culture medium of irradiated cells were also evaluated. IR was capable of down regulating cell surface and total HLA-G levels, with a concomitant increase of sHLA-G1 in the medium. These results could indicate that {gamma}-irradiation decreases HLA-G1 surface levels by enhancing the proteolytic cleavage of this molecule. (authors)

  4. Occupational levels of radiation exposure induce surface expression of interleukin-2 receptors in stimulated human peripheral blood lymphocytes

    International Nuclear Information System (INIS)

    Xu Yindong; Greenstock, C.L.; Trivedi, A.; Mitchel, R.E.J.

    1996-01-01

    Interleukin-2 (IL-2) is a cytokine responsible for a variety of immune and non-immune stimulatory and regulatory functions, including the activation and stimulation of cytotoxic cells able to recognize and kill human tumour cells and T-cell proliferation and differentiation. We show that low doses of radiation, in the range commonly received by atomic radiation workers or as a result of minor medical diagnostic procedures (0.25 to 10 mGy), stimulate the expression of IL-2 receptors (IL-2R) on the surface of peripheral blood lymphocytes (PBL) taken from normal human donors. This stimulated surface expression after in vitro irradiation is an indirect effect, resulting from the secretion into the medium of a soluble factor from the irradiated cells. This factor can also stimulate IL-2R surface expression in unirradiated cells. Consequently, radiation stimulation of IL-2R expression in a large population of PBL shows a triggered-type response rather than being proportional to dose. These results demonstrate that normal human cells can respond to doses of radiation in the range of common occupational or medical exposures. The data also demonstrate a possible defence mechanism against environmental stress by which a radiation-exposed cell can use an indirect signalling mechanism to communicate with and influence the biological processes in an unexposed cell. (orig.). With 1 fig., 4 tabs

  5. S1PR1 Tyr143 phosphorylation downregulates endothelial cell surface S1PR1 expression and responsiveness

    Science.gov (United States)

    Chavez, Alejandra; Schmidt, Tracy Thennes; Yazbeck, Pascal; Rajput, Charu; Desai, Bhushan; Sukriti, Sukriti; Giantsos-Adams, Kristina; Knezevic, Nebojsa; Malik, Asrar B.; Mehta, Dolly

    2015-01-01

    ABSTRACT Activation of sphingosine-1-phosphate receptor 1 (S1PR1) plays a key role in repairing endothelial barrier function. We addressed the role of phosphorylation of the three intracellular tyrosine residues of S1PR1 in endothelial cells in regulating the receptor responsiveness and endothelial barrier function regulated by sphingosine 1-phosphate (S1P)-mediated activation of S1PR1. We demonstrated that phosphorylation of only Y143 site was required for S1PR1 internalization in response to S1P. Maximal S1PR1 internalization was seen in 20 min but S1PR1 returned to the cell surface within 1 h accompanied by Y143-dephosphorylation. Cell surface S1PR1 loss paralleled defective endothelial barrier enhancement induced by S1P. Expression of phospho-defective (Y143F) or phospho-mimicking (Y143D) mutants, respectively, failed to internalize or showed unusually high receptor internalization, consistent with the requirement of Y143 in regulating cell surface S1PR1 expression. Phosphorylation of the five S1PR1 C-terminal serine residues did not affect the role of Y143 phosphorylation in signaling S1PR1 internalization. Thus, rapid reduction of endothelial cell surface expression of S1PR1 subsequent to Y143 phosphorylation is a crucial mechanism of modulating S1PR1 signaling, and hence the endothelial barrier repair function of S1P. PMID:25588843

  6. Effect of nitrogen-rich cell culture surfaces on type X collagen expression by bovine growth plate chondrocytes

    Directory of Open Access Journals (Sweden)

    Wertheimer Michael R

    2011-01-01

    Full Text Available Abstract Background Recent evidence indicates that osteoarthritis (OA may be a systemic disease since mesenchymal stem cells (MSCs from OA patients express type X collagen, a marker of late stage chondrocyte hypertrophy (associated with endochondral ossification. We recently showed that the expression of type X collagen was suppressed when MSCs from OA patients were cultured on nitrogen (N-rich plasma polymer layers, which we call "PPE:N" (N-doped plasma-polymerized ethylene, containing up to 36 atomic percentage (at.% of N. Methods In the present study, we examined the expression of type X collagen in fetal bovine growth plate chondrocytes (containing hypertrophic chondrocytes cultured on PPE:N. We also studied the effect of PPE:N on the expression of matrix molecules such as type II collagen and aggrecan, as well as on proteases (matrix metalloproteinase-13 (MMP-13 and molecules implicated in cell division (cyclin B2. Two other culture surfaces, "hydrophilic" polystyrene (PS, regular culture dishes and nitrogen-containing cation polystyrene (Primaria®, were also investigated for comparison. Results Results showed that type X collagen mRNA levels were suppressed when cultured for 4 days on PPE:N, suggesting that type X collagen is regulated similarly in hypertrophic chondrocytes and in human MSCs from OA patients. However, the levels of type X collagen mRNA almost returned to control value after 20 days in culture on these surfaces. Culture on the various surfaces had no significant effects on type II collagen, aggrecan, MMP-13, and cyclin B2 mRNA levels. Conclusion Hypertrophy is diminished by culturing growth plate chondrocytes on nitrogen-rich surfaces, a mechanism that is beneficial for MSC chondrogenesis. Furthermore, one major advantage of such "intelligent surfaces" over recombinant growth factors for tissue engineering and cartilage repair is potentially large cost-saving.

  7. Developmental expression of a cell surface protein involved in sea urchin skeleton formation. [Strongylocentrotus purpuratus; Lytechinus pictus

    Energy Technology Data Exchange (ETDEWEB)

    Farach, M.C.; Valdizan, M.; Park, H.R.; Decker, G.L.; Lennarz, W.J.

    1986-05-01

    The authors have previously used a monoclonal antibody (1223) to identify a 130 Kd cell surface protein involved in skeleton formation is sea urchin embryos. In the current study the authors have examined the expression of the 1223 antigen over the course of development of embryos of two species, Strongylocentrotus purpuratus and Lytechinus pictus. The 130 Kd protein is detected in S. purp eggs on immunoblots. Labeling with (/sup 3/H) leucine and immunoaffinity chromatography show that it also is synthesized shortly after fertilization. Immunofluroescence reveals that at this early stage the 1223 antigen is uniformly distributed on all of the cells. Synthesis decreases to a minimum by the time of hatching (18 h), as does the total amount of antigen present in the embryo. A second period of synthesis commences at the mesenchyme blastula stage, when the spicule-forming primary mesenchyme cells (PMCs) have appeared. During this later stage, synthesis and cell surface expression are restricted to the PMCs. In contrast to S. purp., in L. pictus the 130 Kd protein does not appear until the PMCs are formed. Hybrid embryos demonstrate a pattern of expression of the maternal species. These results suggest that early expression of 1223 antigen in S. purp. is due to utilization of maternal transcripts present in the egg. In both species later expression in PMCs appears to be the result of cell-type specific synthesis, perhaps encoded by embryonic transcripts.

  8. Kinetoplastid membrane protein-11 is present in promastigotes and amastigotes of Leishmania amazonensis and its surface expression increases during metacyclogenesis

    Directory of Open Access Journals (Sweden)

    Denise CS Matos

    2010-05-01

    Full Text Available Kinetoplastid membrane protein-11 (KMP-11, a protein present in all kinetoplastid protozoa, is considered a potential candidate for a leishmaniasis vaccine. A suitable leishmaniasis vaccine candidate molecule must be expressed in amastigotes, the infective stage for mammals. However, the expression of KMP-11 in Leishmania amastigotes has been a subject of controversy. We evaluated the expression of this molecule in logarithmic and stationary growth phase promastigotes, as well as in amastigotes, of Leishmania amazonensis by immunoblotting, flow cytometry and immunocytochemistry, using a monoclonal antibody against KMP-11. We found that KMP-11 is present in promastigotes and amastigotes. In both stages, the protein was found in association with membrane structures (at the cell surface, flagellar pocket and intracellular vesicles. More importantly, its surface expression is higher in amastigotes than in promastigotes and increases during metacyclogenesis. The increased expression of KMP-11 in metacyclic promastigotes, and especially in amastigotes, indicates a role for this molecule in the parasite relationship with the mammalian host. The presence of this molecule in amastigotes is consistent with the previously demonstrated immunoprotective capacity of vaccine prototypes based on the KMP-11-coding gene and the presence of humoral and cellular immune responses to KMP-11 in Leishmania-infected humans and animals.

  9. The Optimisation of the Expression of Recombinant Surface Immunogenic Protein of Group B Streptococcus in Escherichia coli by Response Surface Methodology Improves Humoral Immunity.

    Science.gov (United States)

    Díaz-Dinamarca, Diego A; Jerias, José I; Soto, Daniel A; Soto, Jorge A; Díaz, Natalia V; Leyton, Yessica Y; Villegas, Rodrigo A; Kalergis, Alexis M; Vásquez, Abel E

    2018-03-01

    Group B Streptococcus (GBS) is the leading cause of neonatal meningitis and a common pathogen in livestock and aquaculture industries around the world. Conjugate polysaccharide and protein-based vaccines are under development. The surface immunogenic protein (SIP) is a conserved protein in all GBS serotypes and has been shown to be a good target for vaccine development. The expression of recombinant proteins in Escherichia coli cells has been shown to be useful in the development of vaccines, and the protein purification is a factor affecting their immunogenicity. The response surface methodology (RSM) and Box-Behnken design can optimise the performance in the expression of recombinant proteins. However, the biological effect in mice immunised with an immunogenic protein that is optimised by RSM and purified by low-affinity chromatography is unknown. In this study, we used RSM for the optimisation of the expression of the rSIP, and we evaluated the SIP-specific humoral response and the property to decrease the GBS colonisation in the vaginal tract in female mice. It was observed by NI-NTA chromatography that the RSM increases the yield in the expression of rSIP, generating a better purification process. This improvement in rSIP purification suggests a better induction of IgG anti-SIP immune response and a positive effect in the decreased GBS intravaginal colonisation. The RSM applied to optimise the expression of recombinant proteins with immunogenic capacity is an interesting alternative in the evaluation of vaccines in preclinical phase, which could improve their immune response.

  10. Elastic Wavespeed Images of Northern Chile Subduction Zone from the Joint Inversion of Body and Surface Waves: Structure of the Andean Forearc and the Double Seismic Zone

    Science.gov (United States)

    Comte, D.; Carrizo, D.; Roecker, S. W.; Peyrat, S.; Arriaza, R.; Chi, R. K.; Baeza, S.

    2015-12-01

    Partly in anticipation of an imminent megathrust earthquake, a significant amount of seismic data has been collected over the past several years in northern Chile by local deployments of seismometers. In this study we generate elastic wavespeed images of the crust and upper mantle using a combination of body wave arrival times and surface wave dispersion curves. The body wave data set consists of 130000 P and 108000 S wave arrival times generated by 12000 earthquakes recorded locally over a period of 25 years by networks comprising about 360 stations. The surface wave data set consists of Rayleigh wave dispersion curves determined from ambient noise recorded by 60 broad band stations from three different networks over a period of three years. Transit time biases due to an uneven distribution of noise were estimated using a technique based on that of Yao and van der Hilst (2009) and found to be as high as 5% for some station pairs. We jointly invert the body and surface wave observations to both improve the overall resolution of the crustal images and reduce the trade-off between shallow and deep structures in the images of the subducted slab. Of particular interest in these images are three regions of anomalous Vp/Vs: (1) An extensive zone of low Vp/Vs (1.68) correlates with trench-parallel magmatic belts emplaced in the upper continental crust. In the region of the coast and continental slope, low Vp/Vs corresponds to batholithic structures in the Jurassic-Cretaceous magmatic arc. Between the central depression and Domeyko Cordillera, low Vp/Vs correlates with the distribution of magmatic arcs of Paleocene-Oligocene and Eocene-Oligocene age. Low Vp/Vs also correlates with the location of the Mejillones Peninsula. (2) A region of high Vp/Vs occurs in what is most likely the serpentinized wedge of the subduction zone. (3) An additional zone of low Vp/Vs is located in the middle of the double seismic zone at depths of 90-110 km. This region may exist all along the

  11. Expression of hepatitis B surface antigen (HBsAg) gene in ...

    African Journals Online (AJOL)

    The transformed nature of the plants was confirmed by polymerase chain reaction (PCR) analysis and Southern blot hybridization. Expression of HBsAg was confirmed by western blotting and levels of expression were assayed by enzyme-linked immunosorbent assay (ELISA). Southern blot hybridization confirmed the ...

  12. Comparison of Gene Expression Programming with neuro-fuzzy and neural network computing techniques in estimating daily incoming solar radiation in the Basque Country (Northern Spain)

    International Nuclear Information System (INIS)

    Landeras, Gorka; López, José Javier; Kisi, Ozgur; Shiri, Jalal

    2012-01-01

    Highlights: ► Solar radiation estimation based on Gene Expression Programming is unexplored. ► This approach is evaluated for the first time in this study. ► Other artificial intelligence models (ANN and ANFIS) are also included in the study. ► New alternatives for solar radiation estimation based on temperatures are provided. - Abstract: Surface incoming solar radiation is a key variable for many agricultural, meteorological and solar energy conversion related applications. In absence of the required meteorological sensors for the detection of global solar radiation it is necessary to estimate this variable. Temperature based modeling procedures are reported in this study for estimating daily incoming solar radiation by using Gene Expression Programming (GEP) for the first time, and other artificial intelligence models such as Artificial Neural Networks (ANNs), and Adaptive Neuro-Fuzzy Inference System (ANFIS). A comparison was also made among these techniques and traditional temperature based global solar radiation estimation equations. Root mean square error (RMSE), mean absolute error (MAE) RMSE-based skill score (SS RMSE ), MAE-based skill score (SS MAE ) and r 2 criterion of Nash and Sutcliffe criteria were used to assess the models’ performances. An ANN (a four-input multilayer perceptron with 10 neurons in the hidden layer) presented the best performance among the studied models (2.93 MJ m −2 d −1 of RMSE). The ability of GEP approach to model global solar radiation based on daily atmospheric variables was found to be satisfactory.

  13. Co-expression of TIMP-1 and its cell surface binding partner CD63 in glioblastomas

    DEFF Research Database (Denmark)

    Aaberg-Jessen, Charlotte; Sørensen, Mia D.; Matos, Ana L.S.A.

    2018-01-01

    , Iba1 did not appear to impact the prognostic value of CD63. A significant correlation was found between TIMP-1 and CD63, and the TIMP-1 and CD63 proteins were co-expressed at the cellular level and located in close molecular proximity, suggesting that TIMP-1 and CD63 could be co-players...... of this study was to assess CD63 expression in astrocytomas focusing on the prognostic potential of CD63 alone and in combination with TIMP-1. Methods: CD63 expression was investigated immunohistochemically in a cohort of 111 astrocytomas and correlated to tumor grade and overall survival by semi-quantitative...... scoring. CD63 expression in tumor-associated microglia/macrophages was examined by double-immunofluorescence with ionized calcium-binding adapter molecule 1 (Iba1). The association between CD63 and TIMP-1 was investigated using previously obtained TIMP-1 data from our astrocytoma cohort. Cellular co-expression...

  14. Surface TRAIL decoy receptor-4 expression is correlated with TRAIL resistance in MCF7 breast cancer cells

    Directory of Open Access Journals (Sweden)

    Aydin Cigdem

    2005-05-01

    Full Text Available Abstract Background Tumor Necrosis Factor (TNF-Related Apoptosis-Inducing Ligand (TRAIL selectively induces apoptosis in cancer cells but not in normal cells. Despite this promising feature, TRAIL resistance observed in cancer cells seriously challenged the use of TRAIL as a death ligand in gene therapy. The current dispute concerns whether or not TRAIL receptor expression pattern is the primary determinant of TRAIL sensitivity in cancer cells. This study investigates TRAIL receptor expression pattern and its connection to TRAIL resistance in breast cancer cells. In addition, a DcR2 siRNA approach and a complementary gene therapy modality involving IKK inhibition (AdIKKβKA were also tested to verify if these approaches could sensitize MCF7 breast cancer cells to adenovirus delivery of TRAIL (Ad5hTRAIL. Methods TRAIL sensitivity assays were conducted using Molecular Probe's Live/Dead Cellular Viability/Cytotoxicity Kit following the infection of breast cancer cells with Ad5hTRAIL. The molecular mechanism of TRAIL induced cell death under the setting of IKK inhibition was revealed by Annexin V binding. Novel quantitative Real Time RT-PCR and flow cytometry analysis were performed to disclose TRAIL receptor composition in breast cancer cells. Results MCF7 but not MDA-MB-231 breast cancer cells displayed strong resistance to adenovirus delivery of TRAIL. Only the combinatorial use of Ad5hTRAIL and AdIKKβKA infection sensitized MCF7 breast cancer cells to TRAIL induced cell death. Moreover, novel quantitative Real Time RT-PCR assays suggested that while the level of TRAIL Decoy Receptor-4 (TRAIL-R4 expression was the highest in MCF7 cells, it was the lowest TRAIL receptor expressed in MDA-MB-231 cells. In addition, conventional flow cytometry analysis demonstrated that TRAIL resistant MCF7 cells exhibited substantial levels of TRAIL-R4 expression but not TRAIL decoy receptor-3 (TRAIL-R3 on surface. On the contrary, TRAIL sensitive MDA-MB-231 cells

  15. Surface TRAIL decoy receptor-4 expression is correlated with TRAIL resistance in MCF7 breast cancer cells

    International Nuclear Information System (INIS)

    Sanlioglu, Ahter D; Dirice, Ercument; Aydin, Cigdem; Erin, Nuray; Koksoy, Sadi; Sanlioglu, Salih

    2005-01-01

    Tumor Necrosis Factor (TNF)-Related Apoptosis-Inducing Ligand (TRAIL) selectively induces apoptosis in cancer cells but not in normal cells. Despite this promising feature, TRAIL resistance observed in cancer cells seriously challenged the use of TRAIL as a death ligand in gene therapy. The current dispute concerns whether or not TRAIL receptor expression pattern is the primary determinant of TRAIL sensitivity in cancer cells. This study investigates TRAIL receptor expression pattern and its connection to TRAIL resistance in breast cancer cells. In addition, a DcR2 siRNA approach and a complementary gene therapy modality involving IKK inhibition (AdIKKβKA) were also tested to verify if these approaches could sensitize MCF7 breast cancer cells to adenovirus delivery of TRAIL (Ad5hTRAIL). TRAIL sensitivity assays were conducted using Molecular Probe's Live/Dead Cellular Viability/Cytotoxicity Kit following the infection of breast cancer cells with Ad5hTRAIL. The molecular mechanism of TRAIL induced cell death under the setting of IKK inhibition was revealed by Annexin V binding. Novel quantitative Real Time RT-PCR and flow cytometry analysis were performed to disclose TRAIL receptor composition in breast cancer cells. MCF7 but not MDA-MB-231 breast cancer cells displayed strong resistance to adenovirus delivery of TRAIL. Only the combinatorial use of Ad5hTRAIL and AdIKKβKA infection sensitized MCF7 breast cancer cells to TRAIL induced cell death. Moreover, novel quantitative Real Time RT-PCR assays suggested that while the level of TRAIL Decoy Receptor-4 (TRAIL-R4) expression was the highest in MCF7 cells, it was the lowest TRAIL receptor expressed in MDA-MB-231 cells. In addition, conventional flow cytometry analysis demonstrated that TRAIL resistant MCF7 cells exhibited substantial levels of TRAIL-R4 expression but not TRAIL decoy receptor-3 (TRAIL-R3) on surface. On the contrary, TRAIL sensitive MDA-MB-231 cells displayed very low levels of surface TRAIL-R4

  16. Projected sea surface temperatures over the 21st century: Changes in the mean, variability and extremes for large marine ecosystem regions of Northern Oceans

    Directory of Open Access Journals (Sweden)

    Michael A. Alexander

    2018-01-01

    Full Text Available Global climate models were used to assess changes in the mean, variability and extreme sea surface temperatures (SSTs in northern oceans with a focus on large marine ecosystems (LMEs adjacent to North America, Europe, and the Arctic Ocean. Results were obtained from 26 models in the Community Model Intercomparison Project Phase 5 (CMIP5 archive and 30 simulations from the National Center for Atmospheric Research Large Ensemble Community Project (CESM-LENS. All of the simulations used the observed greenhouse gas concentrations for 1976–2005 and the RCP8.5 “business as usual” scenario for greenhouse gases through the remainder of the 21st century. In general, differences between models are substantially larger than among the simulations in the CESM-LENS, indicating that the SST changes are more strongly affected by model formulation than internal climate variability. The annual SST trends over 1976–2099 in the 18 LMEs examined here are all positive ranging from 0.05 to 0.5°C decade–1. SST changes by the end of the 21st century are primarily due to a positive shift in the mean with only modest changes in the variability in most LMEs, resulting in a substantial increase in warm extremes and decrease in cold extremes. The shift in the mean is so large that in many regions SSTs during 2070–2099 will 'always' be warmer than the 'warmest' year during 1976–2005. The SST trends are generally stronger in summer than in winter, as greenhouse gas heating is integrated over a much shallower climatological mixed layer depth in summer than in winter, which amplifies the seasonal cycle of SST over the 21st century. In the Arctic, the mean SST and its variability increases substantially during summer, when it is ice free, but not during winter when a thin layer of ice reforms and SSTs remain near the freezing point.

  17. Analysis of the impacts of major anion variations on surface water acidity particularly with regard to conifer harvesting: case studies from Wales and Northern England

    Directory of Open Access Journals (Sweden)

    C. Neal

    1998-01-01

    Full Text Available Data on the water quality of streams draining a range of acidic and acid sensitive, mainly afforested, upland catchments in mid- and north-Wales and northern-England are described to investigate the acidification effects of conifer harvesting in relation to natural variability. Most sites show a large range in pH and major cation and major anion concentrations. The waters draining from the smaller catchments are more acidic and aluminium bearing reflecting a higher proportion of runoff from the acidic soils in each area. However, there is often a less acidic component of runoff under base-flow conditions due to ground-water contributions particularly within the larger streams. Higher concentrations of nitrate occur for sites which have been felled although declines in concentration occur several years after felling. Multiple regression analysis reveals the importance of cation exchange and within catchment acidification associated with sulphate and nitrate generation. Sulphate also has a component associated with weathering but the patterns vary from catchment to catchment. Analysis of the influence of changing anion concentrations associated with tree harvesting reveals that the acidification induced by increases in nitrate can be offset or reversed by the lowering of chloride and sulphate concentrations due to decreased atmospheric scavenging by the vegetation, reduced evapotranspiration and increased surface runoff diluting the acidity generated. It is concluded that contemporary UK forestry guidelines with an emphasis on phased harvesting of catchments over several years and careful harvesting methodologies can alleviate most problems of stream acidification associated with felling activities and in some cases can reverse the acidification pattern.

  18. Prevalence of Veterinary Antibiotics and Antibiotic-Resistant Escherichia coli in the Surface Water of a Livestock Production Region in Northern China

    Science.gov (United States)

    Zhang, Xuelian; Li, Yanxia; Liu, Bei; Wang, Jing; Feng, Chenghong; Gao, Min; Wang, Lina

    2014-01-01

    This study investigated the occurrence of 12 veterinary antibiotics (VAs) and the susceptibility of Escherichia coli (E. coli) in a rural water system that was affected by livestock production in northern China. Each of the surveyed sites was determined with at least eight antibiotics with maximum concentration of up to 450 ng L−1. The use of VAs in livestock farming probably was a primary source of antibiotics in the rivers. Increasing total antibiotics were measured from up- to mid- and downstream in the two tributaries. Eighty-eight percent of the 218 E. coli isolates that were derived from the study area exhibited, in total, 48 resistance profiles against the eight examined drugs. Significant correlations were found among the resistance rates of sulfamethoxazole-trimethoprim, chloromycetin and ampicillin as well as between tetracycline and chlortetracycline, suggesting a possible cross-selection for resistance among these drugs. The E. coli resistance frequency also increased from up- to midstream in the three rivers. E. coli isolates from different water systems showed varying drug numbers of resistance. No clear relationship was observed in the antibiotic resistance frequency with corresponding antibiotic concentration, indicating that the antibiotic resistance for E. coli in the aquatic environment might be affected by factors besides antibiotics. High numbers of resistant E. coli were also isolated from the conserved reservoir. These results suggest that rural surface water may become a large pool of VAs and resistant bacteria. This study contributes to current information on VAs and resistant bacteria contamination in aquatic environments particularly in areas under intensive agriculture. Moreover, this study indicates an urgent need to monitor the use of VAs in animal production, and to control the release of animal-originated antibiotics into the environment. PMID:25372873

  19. Prevalence of veterinary antibiotics and antibiotic-resistant Escherichia coli in the surface water of a livestock production region in northern China.

    Science.gov (United States)

    Zhang, Xuelian; Li, Yanxia; Liu, Bei; Wang, Jing; Feng, Chenghong; Gao, Min; Wang, Lina

    2014-01-01

    This study investigated the occurrence of 12 veterinary antibiotics (VAs) and the susceptibility of Escherichia coli (E. coli) in a rural water system that was affected by livestock production in northern China. Each of the surveyed sites was determined with at least eight antibiotics with maximum concentration of up to 450 ng L(-1). The use of VAs in livestock farming probably was a primary source of antibiotics in the rivers. Increasing total antibiotics were measured from up- to mid- and downstream in the two tributaries. Eighty-eight percent of the 218 E. coli isolates that were derived from the study area exhibited, in total, 48 resistance profiles against the eight examined drugs. Significant correlations were found among the resistance rates of sulfamethoxazole-trimethoprim, chloromycetin and ampicillin as well as between tetracycline and chlortetracycline, suggesting a possible cross-selection for resistance among these drugs. The E. coli resistance frequency also increased from up- to midstream in the three rivers. E. coli isolates from different water systems showed varying drug numbers of resistance. No clear relationship was observed in the antibiotic resistance frequency with corresponding antibiotic concentration, indicating that the antibiotic resistance for E. coli in the aquatic environment might be affected by factors besides antibiotics. High numbers of resistant E. coli were also isolated from the conserved reservoir. These results suggest that rural surface water may become a large pool of VAs and resistant bacteria. This study contributes to current information on VAs and resistant bacteria contamination in aquatic environments particularly in areas under intensive agriculture. Moreover, this study indicates an urgent need to monitor the use of VAs in animal production, and to control the release of animal-originated antibiotics into the environment.

  20. Prevalence of veterinary antibiotics and antibiotic-resistant Escherichia coli in the surface water of a livestock production region in northern China.

    Directory of Open Access Journals (Sweden)

    Xuelian Zhang

    Full Text Available This study investigated the occurrence of 12 veterinary antibiotics (VAs and the susceptibility of Escherichia coli (E. coli in a rural water system that was affected by livestock production in northern China. Each of the surveyed sites was determined with at least eight antibiotics with maximum concentration of up to 450 ng L(-1. The use of VAs in livestock farming probably was a primary source of antibiotics in the rivers. Increasing total antibiotics were measured from up- to mid- and downstream in the two tributaries. Eighty-eight percent of the 218 E. coli isolates that were derived from the study area exhibited, in total, 48 resistance profiles against the eight examined drugs. Significant correlations were found among the resistance rates of sulfamethoxazole-trimethoprim, chloromycetin and ampicillin as well as between tetracycline and chlortetracycline, suggesting a possible cross-selection for resistance among these drugs. The E. coli resistance frequency also increased from up- to midstream in the three rivers. E. coli isolates from different water systems showed varying drug numbers of resistance. No clear relationship was observed in the antibiotic resistance frequency with corresponding antibiotic concentration, indicating that the antibiotic resistance for E. coli in the aquatic environment might be affected by factors besides antibiotics. High numbers of resistant E. coli were also isolated from the conserved reservoir. These results suggest that rural surface water may become a large pool of VAs and resistant bacteria. This study contributes to current information on VAs and resistant bacteria contamination in aquatic environments particularly in areas under intensive agriculture. Moreover, this study indicates an urgent need to monitor the use of VAs in animal production, and to control the release of animal-originated antibiotics into the environment.

  1. Mucosal Immunogenicity of Genetically Modified Lactobacillus acidophilus Expressing an HIV-1 Epitope within the Surface Layer Protein.

    Directory of Open Access Journals (Sweden)

    Akinobu Kajikawa

    Full Text Available Surface layer proteins of probiotic lactobacilli are theoretically efficient epitope-displaying scaffolds for oral vaccine delivery due to their high expression levels and surface localization. In this study, we constructed genetically modified Lactobacillus acidophilus strains expressing the membrane proximal external region (MPER from human immunodeficiency virus type 1 (HIV-1 within the context of the major S-layer protein, SlpA. Intragastric immunization of mice with the recombinants induced MPER-specific and S-layer protein-specific antibodies in serum and mucosal secretions. Moreover, analysis of systemic SlpA-specific cytokines revealed that the responses appeared to be Th1 and Th17 dominant. These findings demonstrated the potential use of the Lactobacillus S-layer protein for development of oral vaccines targeting specific peptides.

  2. Mucosal Immunogenicity of Genetically Modified Lactobacillus acidophilus Expressing an HIV-1 Epitope within the Surface Layer Protein

    Science.gov (United States)

    Kajikawa, Akinobu; Zhang, Lin; LaVoy, Alora; Bumgardner, Sara; Klaenhammer, Todd R.; Dean, Gregg A.

    2015-01-01

    Surface layer proteins of probiotic lactobacilli are theoretically efficient epitope-displaying scaffolds for oral vaccine delivery due to their high expression levels and surface localization. In this study, we constructed genetically modified Lactobacillus acidophilus strains expressing the membrane proximal external region (MPER) from human immunodeficiency virus type 1 (HIV-1) within the context of the major S-layer protein, SlpA. Intragastric immunization of mice with the recombinants induced MPER-specific and S-layer protein-specific antibodies in serum and mucosal secretions. Moreover, analysis of systemic SlpA-specific cytokines revealed that the responses appeared to be Th1 and Th17 dominant. These findings demonstrated the potential use of the Lactobacillus S-layer protein for development of oral vaccines targeting specific peptides. PMID:26509697

  3. Pectinesterase Inhibitor from Jelly Fig (Ficus awkeotsang Makino Achene Inhibits Surface Antigen Expression by Human Hepatitis B Virus

    Directory of Open Access Journals (Sweden)

    Yu-Chuen Huang

    2013-01-01

    Full Text Available Pectinesterase inhibitor (PEI isolated from jelly fig (Ficus awkeotsang Makino is an edible component of a popular drink consumed in Asia. Hepatitis B virus (HBV infection is prevalent in Asia, and current treatments for HBV infection need improvement. This study aimed to evaluate the effect of PEI on the surface antigen expression by HBV (HBsAg. Human hepatoma cell lines Hep3B and Huh7 served as in vitro models for assessing the cytotoxicity and HBsAg expression. A culture of primary hepatocytes cultured from mice served as the normal counterpart. Cell viability was measured by 3-(4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide (MTT colorimetric assay. HBsAg expression was evaluated by measuring HBsAg secretion into the culture medium using an enzyme-linked immunosorbent assay. The results showed that PEI did not affect the viability of the human hepatoma cell lines or primary mouse hepatocytes. PEI inhibited the expression of HBsAg in hepatoma cell lines harboring endogenous (Hep3B and integrated (Huh7 HBV genomes in a concentration- and time-dependent manner, thus implicating a universal activity against HBV gene expression. In conclusion, it suggests that PEI from jelly fig inhibits the expression of human HBsAg in host cells without toxic effects on normal primary hepatocytes.

  4. Pectinesterase Inhibitor from Jelly Fig (Ficus awkeotsang Makino) Achene Inhibits Surface Antigen Expression by Human Hepatitis B Virus.

    Science.gov (United States)

    Huang, Yu-Chuen; Jiang, Chii-Ming; Chen, Yu-Jen; Chen, Yu-Yawn

    2013-01-01

    Pectinesterase inhibitor (PEI) isolated from jelly fig (Ficus awkeotsang Makino) is an edible component of a popular drink consumed in Asia. Hepatitis B virus (HBV) infection is prevalent in Asia, and current treatments for HBV infection need improvement. This study aimed to evaluate the effect of PEI on the surface antigen expression by HBV (HBsAg). Human hepatoma cell lines Hep3B and Huh7 served as in vitro models for assessing the cytotoxicity and HBsAg expression. A culture of primary hepatocytes cultured from mice served as the normal counterpart. Cell viability was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric assay. HBsAg expression was evaluated by measuring HBsAg secretion into the culture medium using an enzyme-linked immunosorbent assay. The results showed that PEI did not affect the viability of the human hepatoma cell lines or primary mouse hepatocytes. PEI inhibited the expression of HBsAg in hepatoma cell lines harboring endogenous (Hep3B) and integrated (Huh7) HBV genomes in a concentration- and time-dependent manner, thus implicating a universal activity against HBV gene expression. In conclusion, it suggests that PEI from jelly fig inhibits the expression of human HBsAg in host cells without toxic effects on normal primary hepatocytes.

  5. 2-deoxy D-glucose prevents cell surface expression of NKG2D ligands through inhibition of N-linked glycosylation

    DEFF Research Database (Denmark)

    Andresen, Lars; Skovbakke, Sarah Line; Persson, Gry

    2012-01-01

    NKG2D ligand surface expression is important for immune recognition of stressed and neotransformed cells. In this study, we show that surface expression of MICA/B and other NKG2D ligands is dependent on N-linked glycosylation. The inhibitor of glycolysis and N-linked glycosylation, 2-deoxy...

  6. Generation of murine tumor cell lines deficient in MHC molecule surface expression using the CRISPR/Cas9 system.

    Science.gov (United States)

    Das, Krishna; Eisel, David; Lenkl, Clarissa; Goyal, Ashish; Diederichs, Sven; Dickes, Elke; Osen, Wolfram; Eichmüller, Stefan B

    2017-01-01

    In this study, the CRISPR/Cas9 technology was used to establish murine tumor cell lines, devoid of MHC I or MHC II surface expression, respectively. The melanoma cell line B16F10 and the murine breast cancer cell line EO-771, the latter stably expressing the tumor antigen NY-BR-1 (EO-NY), were transfected with an expression plasmid encoding a β2m-specific single guide (sg)RNA and Cas9. The resulting MHC I negative cells were sorted by flow cytometry to obtain single cell clones, and loss of susceptibility of peptide pulsed MHC I negative clones to peptide-specific CTL recognition was determined by IFNγ ELISpot assay. The β2m knockout (KO) clones did not give rise to tumors in syngeneic mice (C57BL/6N), unless NK cells were depleted, suggesting that outgrowth of the β2m KO cell lines was controlled by NK cells. Using sgRNAs targeting the β-chain encoding locus of the IAb molecule we also generated several B16F10 MHC II KO clones. Peptide loaded B16F10 MHC II KO cells were insusceptible to recognition by OT-II cells and tumor growth was unaltered compared to parental B16F10 cells. Thus, in our hands the CRISPR/Cas9 system has proven to be an efficient straight forward strategy for the generation of MHC knockout cell lines. Such cell lines could serve as parental cells for co-transfection of compatible HLA alleles together with human tumor antigens of interest, thereby facilitating the generation of HLA matched transplantable tumor models, e.g. in HLAtg mouse strains of the newer generation, lacking cell surface expression of endogenous H2 molecules. In addition, our tumor cell lines established might offer a useful tool to investigate tumor reactive T cell responses that function independently from MHC molecule surface expression by the tumor.

  7. Generation of murine tumor cell lines deficient in MHC molecule surface expression using the CRISPR/Cas9 system.

    Directory of Open Access Journals (Sweden)

    Krishna Das

    Full Text Available In this study, the CRISPR/Cas9 technology was used to establish murine tumor cell lines, devoid of MHC I or MHC II surface expression, respectively. The melanoma cell line B16F10 and the murine breast cancer cell line EO-771, the latter stably expressing the tumor antigen NY-BR-1 (EO-NY, were transfected with an expression plasmid encoding a β2m-specific single guide (sgRNA and Cas9. The resulting MHC I negative cells were sorted by flow cytometry to obtain single cell clones, and loss of susceptibility of peptide pulsed MHC I negative clones to peptide-specific CTL recognition was determined by IFNγ ELISpot assay. The β2m knockout (KO clones did not give rise to tumors in syngeneic mice (C57BL/6N, unless NK cells were depleted, suggesting that outgrowth of the β2m KO cell lines was controlled by NK cells. Using sgRNAs targeting the β-chain encoding locus of the IAb molecule we also generated several B16F10 MHC II KO clones. Peptide loaded B16F10 MHC II KO cells were insusceptible to recognition by OT-II cells and tumor growth was unaltered compared to parental B16F10 cells. Thus, in our hands the CRISPR/Cas9 system has proven to be an efficient straight forward strategy for the generation of MHC knockout cell lines. Such cell lines could serve as parental cells for co-transfection of compatible HLA alleles together with human tumor antigens of interest, thereby facilitating the generation of HLA matched transplantable tumor models, e.g. in HLAtg mouse strains of the newer generation, lacking cell surface expression of endogenous H2 molecules. In addition, our tumor cell lines established might offer a useful tool to investigate tumor reactive T cell responses that function independently from MHC molecule surface expression by the tumor.

  8. Actual Evapotranspiration in the Al-Khazir Gomal Basin (Northern Iraq Using the Surface Energy Balance Algorithm for Land (SEBAL and Water Balance

    Directory of Open Access Journals (Sweden)

    Hussein Jassas

    2015-04-01

    Full Text Available Increasing dependence on groundwater requires a detailed determination of the different outputs and inputs of a basin for better water management. Determination of spatial and temporal actual evapotranspiration (ETa, in this regard, is of vital importance as there is significant water loss from drainage basins. This research paper uses the Surface Energy Balance Algorithm for Land (SEBAL, as well as the water balance, to estimate the spatial and temporal ETa in the Al-Khazir Gomal Basin, Northern Iraq. To compensate for the shortage in rainfall, and to irrigate summer crops, farmers in this basin have been depending, to a large extent, on groundwater extracted from the underlying unconfined aquifer, which is considered the major source for both domestic and agricultural uses in this basin. Rainfed farming of wheat and barley is one of the most important activities in the basin in the winter season, while in the summer season, agricultural activity is limited to small rice fields and narrow strips of vegetable cultivation along the Al-Khazir River. The Landsat Thematic Mapper images (TM5 acquired on 21 November 2006, 9 March 2007, 5 May 2007, 21 July 2007, and 23 September 2007 were used, along with a digital elevation model (DEM and ground-based meteorological data, measured within the area of interest. Estimation of seasonal ETa from periods between satellite overpasses was computed using the evaporative fraction (Ʌ. The water balance approach was utilized, using meteorological data and river hydrograph analysis, to estimate the ETa as the only missing input in the predefined water balance equation. The results of the two applied methods were comparable. SEBAL results were compared with the land use land cover (LULC map. The river showed the highest ETa, as evaporation from the free-water surface. Rice fields, irrigated in the summer season, have a high ETa in the images, as these fields are immersed in water during June, July and August

  9. Development of a flagellin surface display expression system in a moderate thermophile, Bacillus halodurans Alk36

    CSIR Research Space (South Africa)

    Crampton, Michael C

    2007-06-01

    Full Text Available inserted as in-frame, chimeric, flagellin sandwich fusions to identify the permissive insertion sites corresponding to the variable regions of the flagellin protein. Flagellin expression and motility were evaluated for these constructs. Two sites were...

  10. Platelet surface expression of SDF-1 is associated with clinical outcomes in the patients with cardiovascular disease.

    Science.gov (United States)

    Rath, Dominik; Chatterjee, Madhumita; Bongartz, Angela; Müller, Karin; Droppa, Michal; Stimpfle, Fabian; Borst, Oliver; Zuern, Christine; Vogel, Sebastian; Gawaz, Meinrad; Geisler, Tobias

    2017-01-01

    Platelet surface expression levels of stromal cell derived factor 1 (SDF-1) are elevated in acute coronary syndrome and associated with LVEF% improvement after myocardial infarction (MI). Platelet SDF-1 might facilitate thrombus formation and endomyocardial expression of SDF-1 is enhanced in inflammatory cardiomyopathy and positively correlates with myocardial fibrosis. The influence of platelet SDF-1 on outcome in the patients with symptomatic coronary artery disease (CAD) is to the best of our knowledge unknown. Blood samples of 608 consecutive CAD patients were collected during the percutaneous coronary intervention and analyzed for surface expression of SDF-1 by flow cytometry. The primary combined endpoint was defined as the composite of either MI, or ischemic stroke, or all-cause death. Secondary endpoints were defined as the aforementioned single events. The patients with baseline platelet SDF-1 levels above the third quartile showed a significantly worse cumulative event-free survival when compared to the patients with lower baseline SDF-1 levels (first to third quartile) (log rank 0.009 for primary combined endpoint and log rank 0.016 for secondary endpoint all-cause death). Multivariate Cox regression analysis showed that SDF-1 levels above the third quartile were independently associated with the primary combined endpoint and the secondary endpoint all-cause death. We provide first clinical evidence that high platelet expression levels of SDF-1 influence clinical outcomes in CAD patients in a negative way.

  11. Cell-surface serglycin promotes adhesion of myeloma cells to collagen type I and affects the expression of matrix metalloproteinases.

    Science.gov (United States)

    Skliris, Antonis; Labropoulou, Vassiliki T; Papachristou, Dionysios J; Aletras, Alexios; Karamanos, Nikos K; Theocharis, Achilleas D

    2013-05-01

    Serglycin (SG) is mainly expressed by hematopoetic cells as an intracellular proteoglycan. Multiple myeloma cells constitutively secrete SG, which is also localized on the cell surface in some cell lines. In this study, SG isolated from myeloma cells was found to interact with collagen type I (Col I), which is a major bone matrix component. Notably, myeloma cells positive for cell-surface SG (csSG) adhered significantly to Col I, compared to cells lacking csSG. Removal of csSG by treatment of the cells with chondroitinase ABC or blocking of csSG by an SG-specific polyclonal antibody significantly reduced the adhesion of myeloma cells to Col I. Significant up-regulation of expression of the matrix metalloproteinases MMP-2 and MMP-9 at both the mRNA and protein levels was observed when culturing csSG-positive myeloma cells on Col I-coated dishes or in the presence of soluble Col I. MMP-9 and MMP-2 were also expressed in increased amounts by myeloma cells in the bone marrow of patients with multiple myeloma. Our data indicate that csSG of myeloma cells affects key functional properties, such as adhesion to Col I and the expression of MMPs, and imply that csSG may serve as a potential prognostic factor and/or target for pharmacological interventions in multiple myeloma. © 2013 The Authors Journal compilation © 2013 FEBS.

  12. Lipopolysaccharide-induced expression of cell surface receptors and cell activation of neutrophils and monocytes in whole human blood

    Directory of Open Access Journals (Sweden)

    N.E. Gomes

    2010-09-01

    Full Text Available Lipopolysaccharide (LPS activates neutrophils and monocytes, inducing a wide array of biological activities. LPS rough (R and smooth (S forms signal through Toll-like receptor 4 (TLR4, but differ in their requirement for CD14. Since the R-form LPS can interact with TLR4 independent of CD14 and the differential expression of CD14 on neutrophils and monocytes, we used the S-form LPS from Salmonella abortus equi and the R-form LPS from Salmonella minnesota mutants to evaluate LPS-induced activation of human neutrophils and monocytes in whole blood from healthy volunteers. Expression of cell surface receptors and reactive oxygen species (ROS and nitric oxide (NO generation were measured by flow cytometry in whole blood monocytes and neutrophils. The oxidative burst was quantified by measuring the oxidation of 2',7'-dichlorofluorescein diacetate and the NO production was quantified by measuring the oxidation of 4-amino-5-methylamino-2',7'-difluorofluorescein diacetate. A small increase of TLR4 expression by monocytes was observed after 6 h of LPS stimulation. Monocyte CD14 modulation by LPS was biphasic, with an initial 30% increase followed by a 40% decrease in expression after 6 h of incubation. Expression of CD11b was rapidly up-regulated, doubling after 5 min on monocytes, while down-regulation of CXCR2 was observed on neutrophils, reaching a 50% reduction after 6 h. LPS induced low production of ROS and NO. This study shows a complex LPS-induced cell surface receptor modulation on human monocytes and neutrophils, with up- and down-regulation depending on the receptor. R- and S-form LPS activate human neutrophils similarly, despite the low CD14 expression, if the stimulation occurs in whole blood.

  13. Plasmodium falciparum parasites expressing pregnancy-specific variant surface antigens adhere strongly to the choriocarcinoma cell line BeWo

    DEFF Research Database (Denmark)

    Haase, Rikke N; Megnekou, Rosette; Lundquist, Maja

    2006-01-01

    Placenta-sequestering Plasmodium falciparum parasites causing pregnancy-associated malaria express pregnancy-specific variant surface antigens (VSA(PAM)). We report here that VSA(PAM)-expressing patient isolates adhere strongly to the choriocarcinoma cell line BeWo and that the BeWo line can...... be used to efficiently select for VSA(PAM) expression in vitro....

  14. Observational evidence of mesoscale variability of the Northern Current (North-Western Mediterranean Sea): a combined study via gliders, HF RADAR, surface drifters, and vessel data

    Science.gov (United States)

    Bellomo, Lucio; Berta, Maristella; Pietro Gasparini, Gian; Griffa, Annalisa; Gatimu Magaldi, Marcello; Marmain, Julien; Molcard, Anne; Vetrano, Anna; Béguery, Laurent; Borghini, Mireno

    2013-04-01

    Results from a combined observational effort put forth in December 2011 are here presented. The focus is on the mesoscale variability of the Northern Current (NC), the branch of the general North-Western Mediterranean cyclonic circulation extending from the Ligurian to the Catalan Sea (Albérola et al., Oceanologica Acta, vol. 18, n. 2, 1995). The study area, located between the Ligurian Sea and the Gulf of Lions, includes the part of French coast between Nice and Toulon, where only a few hydrographic data have been collected in the past. Dynamic instabilities of the NC, observed and reported in literature (Picco et al., Ocean Science, vol. 6, 2010), make this region particularly important, with consequences in the recirculation of the Ligurian Gyre and in the NC intrusions in the Gulf of Lions (Millot and Wald, Oceanologica Acta, vol. 3, n. 4, 1980). This works aims at providing experimental evidence of the effects that mesoscale exerts on the NC dynamics via an innovative and complementary data set. Two Slocum Gliders (a Shallow and a Deep one), both equipped with CTD and dissolved oxygen sensors, sampled the area within 70 km from the coast for about 20 days. The shallow one (200 m) realized six transects describing a "W"-shaped pattern from Nice to Toulon, whereas the deep one (1000 m) performed repeated cross-current sections off Toulon. Concurrent observations were obtained via: a) CTD and both Lowered and Vessel-Mounted ADCP transects obtained during a 5-day oceanographic cruise on board of the Research Vessel Urania; b) repeated deployments of surface drifters; c) a continuously-recording High Frequency (HF) RADAR which measures surface currents off Toulon in a 40 × 25 km2 region with high resolution both in space (2 km) and in time (1 hour). The combined use of data from the shallow glider and the ship-based ADCP measurements reveals the presence of an instability of the offshore front of the NC. Its location is confirmed by high-resolution satellite

  15. Heterologous expression of carcinoembryonic antigen in Lactococcus lactis via LcsB-mediated surface displaying system for oral vaccine development.

    Science.gov (United States)

    Zhang, Xiaowei; Hu, Shumin; Du, Xue; Li, Tiejun; Han, Lanlan; Kong, Jian

    2016-12-01

    Carcinoembryonic antigen (CEA) is an attractive target for immunotherapy because it is expressed minimally in normal tissue, but is overexpressed in a wide variety of malignant epithelial tissues. Lactic acid bacteria (LABs), widely used in food processes, are attractive candidates for oral vaccination. Thus, we examined whether LABs could be used as a live vaccine vector to deliver CEA antigen. CEA was cloned into an Escherichia coli/Lactococcus lactis shuttle vector pSEC:LEISS under the control of a nisin promoter. For displaying the CEA on the cell surface of the L. lactis strain, the anchor motif LcsB from the S-layer protein of Lactobacillus crispatus was fused with CEA. Intracellular and cell surface expression of the CEA-LcsB fusion was confirmed by western blot analysis. Significantly higher levels of CEA-specific secretory immunoglobulin A in the sera of mice were observed upon oral administration of strain cultures containing the CEA-LcsB fused protein. In addition, the CEA-LcsB antigen group showed a higher spleen index compared to the CEA antigen alone or negative control, demonstrating that surface-displayed CEA antigen could induce a higher immune response. These results provided the first evidence for displaying CEA antigen on the cell surfaces of LABs as oral vaccines against cancer or infectious diseases. Copyright © 2014. Published by Elsevier B.V.

  16. Chemistry of Frozen NaCl and MgSO4 Brines - Implications for Surface Expression of Europa's Ocean Composition

    Science.gov (United States)

    Johnson, P. V.; Hodyss, R. P.; Choukroun, M.; Vu, T. H.

    2015-12-01

    The composition of Europa's subsurface ocean is a critical determinant of its habitability, but current analysis of the ocean composition is limited to its expression on the Europan surface. While there is observational evidence indicating that ocean materials make their way to the surface, our understanding of the chemical processes that can alter this material under Europan surface conditions is limited. We present experimental data on the chemistry of mixed solutions of NaCl and MgSO4 as they are frozen to 100 K, replicating the conditions that may occur when subsurface ocean fluids are emplaced onto Europa's surface. Confocal micro-Raman spectroscopy is used to study the formation of salts during the freezing process, and the interaction of ions in the frozen brines. Our data indicate that mixed aqueous solutions of NaCl and MgSO4 form Na2SO4 and MgCl2 preferentially when frozen, rather than making NaCl and MgSO4 precipitates. The detection of epsomite (MgSO4Ÿ•7H2O) on Europa's surface may therefore imply an ocean composition relatively low in sodium, unless radiolytic chemistry converts MgCl2 to MgSO4 as suggested by Hand and Brown 2013 (ApJ 145 110). These results have important implications for the interpretation of remote sensing data of Europa's surface.

  17. Expression robust 3D face recognition via mesh-based histograms of multiple order surface differential quantities

    KAUST Repository

    Li, Huibin

    2011-09-01

    This paper presents a mesh-based approach for 3D face recognition using a novel local shape descriptor and a SIFT-like matching process. Both maximum and minimum curvatures estimated in the 3D Gaussian scale space are employed to detect salient points. To comprehensively characterize 3D facial surfaces and their variations, we calculate weighted statistical distributions of multiple order surface differential quantities, including histogram of mesh gradient (HoG), histogram of shape index (HoS) and histogram of gradient of shape index (HoGS) within a local neighborhood of each salient point. The subsequent matching step then robustly associates corresponding points of two facial surfaces, leading to much more matched points between different scans of a same person than the ones of different persons. Experimental results on the Bosphorus dataset highlight the effectiveness of the proposed method and its robustness to facial expression variations. © 2011 IEEE.

  18. Pancreatic hormones are expressed on the surfaces of human and rat islet cells through exocytotic sites

    DEFF Research Database (Denmark)

    Larsson, L I; Hutton, J C; Madsen, O D

    1989-01-01

    . Electron microscopy reveals the labeling to occur at sites of exocytotic granule release, involving the surfaces of extruded granule cores. The surfaces of islet cells were labeled both by polyclonal and monoclonal antibodies, excluding that receptor-interacting, anti-idiotypic hormone antibodies were...... for these results. It is concluded that the staining reflects interactions between the appropriate antibodies and exocytotic sites of hormone release....

  19. Response surface studies that elucidate the role of infiltration conditions on Agrobacterium tumefaciens-mediated transient transgene expression in harvested switchgrass (Panicum virgatum)

    Energy Technology Data Exchange (ETDEWEB)

    VanderGheynst, J.S.; Guo, H.-Y.; Simmons, C.W. [Department of Biological and Agricultural Engineering, University of California, Davis, One Shields Avenue, Davis, CA 95616 (United States)

    2008-04-15

    Agrobacterium tumefaciens-mediated transient expression (agroinfiltration) experiments were performed in harvested switchgrass (Panicum virgatum) leaves to identify the effects of wounding by bead beating, surfactant concentration and vacuum application on in planta {beta}-glucuronidase expression and leaf decay. Expression was scored based on a consistent pattern of visual observations of histochemical staining over the leaf surface as might be observed in stable gene expression in switchgrass leaves. Assays on extracts from leaves were also performed to measure expression levels; however, these assays showed low expression levels, which may have been due to low recombinant protein recovery and decomposition in the leaf. Bead beating was successful for wounding the plant surface, but did not improve the consistency of expression based on histochemical staining observations. Surfactant was necessary for improving contact between the leaf surface and Agrobacterium suspension and consistently improved expression when vacuum application level was low (25 kPa). Increasing vacuum application from 25 to 5 kPa improved expression only when surfactant concentration was low. When a suspension of A. tumefaciens containing 1000 ppm Break-Thru surfactant was added to harvested leaves and 25 kPa vacuum applied, a fairly uniform expression was visualized across the leaf surface within 2-3 days of incubation, suggesting that agroinfiltration is a rapid tool for examining expression of transgenes in switchgrass leaves. (author)

  20. Determining Cell-surface Expression and Endocytic Rate of Proteins in Primary Astrocyte Cultures Using Biotinylation.

    Science.gov (United States)

    Tham, Daniel Kai Long; Moukhles, Hakima

    2017-07-03

    Cell-surface proteins mediate a wide array of functions. In many cases, their activity is regulated by endocytic processes that modulate their levels at the plasma membrane. Here, we present detailed protocols for 2 methods that facilitate the study of such processes, both of which are based on the principle of the biotinylation of cell-surface proteins. The first is designed to allow for the semi-quantitative determination of the relative levels of a particular protein at the cell-surface. In it, the lysine residues of the plasma membrane proteins of cells are first labeled with a biotin moiety. Once the cells are lysed, these proteins may then be specifically precipitated via the use of agarose-immobilized streptavidin by exploiting the natural affinity of the latter for biotin. The proteins isolated in such a manner may then be analyzed via a standard western blotting approach. The second method provides a means of determining the endocytic rate of a particular cell-surface target over a period of time. Cell-surface proteins are first modified with a biotin derivative containing a cleavable disulfide bond. The cells are then shifted back to normal culture conditions, which causes the endocytic uptake of a proportion of biotinylated proteins. Next, the disulfide bonds of non-internalized biotin groups are reduced using the membrane-impermeable reducing agent glutathione. Via this approach, endocytosed proteins may thus be isolated and quantified with a high degree of specificity.

  1. Gang Identity or Self-Expression? Researchers Look beyond the Surface of "Gang Clothing" and Appearance.

    Science.gov (United States)

    Hethorn, Janet

    1994-01-01

    A study that included observations and interviews in street and school settings throughout California and in selected cities nationwide documented the changing nature of gang identity within the broader context of street style and adolescent expression. Suggests that targeting student behavior, as opposed to style of clothing, would be more…

  2. Colonic epithelial cell expression of ICAM-1 relates to loss of surface continuity

    DEFF Research Database (Denmark)

    Vainer, Ben; Horn, Thomas; Nielsen, Ole Haagen

    2006-01-01

    OBJECTIVE: Intercellular adhesion molecule-1 (ICAM-1) is important in ulcerative colitis (UC) by mediating the arrest and further migration of neutrophils. In vitro studies have shown that colonocytes from chronically inflamed colon and cultured colon cancer cells are capable of expressing ICAM-1...

  3. Surface expression and limited proteolysis of ADAM10 are increased by a dominant negative inhibitor of dynamin

    Directory of Open Access Journals (Sweden)

    Slack Barbara E

    2011-05-01

    Full Text Available Abstract Background The amyloid precursor protein (APP is cleaved by β- and γ-secretases to generate toxic amyloid β (Aβ peptides. Alternatively, α-secretases cleave APP within the Aβ domain, precluding Aβ formation and releasing the soluble ectodomain, sAPPα. We previously showed that inhibition of the GTPase dynamin reduced APP internalization and increased release of sAPPα, apparently by prolonging the interaction between APP and α-secretases at the plasma membrane. This was accompanied by a reduction in Aβ generation. In the present study, we investigated whether surface expression of the α-secretase ADAM (a disintegrin and metalloprotease10 is also regulated by dynamin-dependent endocytosis. Results Transfection of human embryonic kidney (HEK cells stably expressing M3 muscarinic receptors with a dominant negative dynamin I mutant (dyn I K44A, increased surface expression of both immature, and mature, catalytically active forms of co-expressed ADAM10. Surface levels of ADAM10 were unaffected by activation of protein kinase C (PKC or M3 receptors, indicating that receptor-coupled shedding of the ADAM substrate APP is unlikely to be mediated by inhibition of ADAM10 endocytosis in this cell line. Dyn I K44A strongly increased the formation of a C-terminal fragment of ADAM10, consistent with earlier reports that the ADAM10 ectodomain is itself a target for sheddases. The abundance of this fragment was increased in the presence of a γ-secretase inhibitor, but was not affected by M3 receptor activation. The dynamin mutant did not affect the distribution of ADAM10 and its C-terminal fragment between raft and non-raft membrane compartments. Conclusions Surface expression and limited proteolysis of ADAM10 are regulated by dynamin-dependent endocytosis, but are unaffected by activation of signaling pathways that upregulate shedding of ADAM substrates such as APP. Modulation of ADAM10 internalization could affect cellular behavior in two

  4. Surface expression and limited proteolysis of ADAM10 are increased by a dominant negative inhibitor of dynamin.

    Science.gov (United States)

    Carey, Robyn M; Blusztajn, Jan K; Slack, Barbara E

    2011-05-17

    The amyloid precursor protein (APP) is cleaved by β- and γ-secretases to generate toxic amyloid β (Aβ) peptides. Alternatively, α-secretases cleave APP within the Aβ domain, precluding Aβ formation and releasing the soluble ectodomain, sAPPα. We previously showed that inhibition of the GTPase dynamin reduced APP internalization and increased release of sAPPα, apparently by prolonging the interaction between APP and α-secretases at the plasma membrane. This was accompanied by a reduction in Aβ generation. In the present study, we investigated whether surface expression of the α-secretase ADAM (a disintegrin and metalloprotease)10 is also regulated by dynamin-dependent endocytosis. Transfection of human embryonic kidney (HEK) cells stably expressing M3 muscarinic receptors with a dominant negative dynamin I mutant (dyn I K44A), increased surface expression of both immature, and mature, catalytically active forms of co-expressed ADAM10. Surface levels of ADAM10 were unaffected by activation of protein kinase C (PKC) or M3 receptors, indicating that receptor-coupled shedding of the ADAM substrate APP is unlikely to be mediated by inhibition of ADAM10 endocytosis in this cell line. Dyn I K44A strongly increased the formation of a C-terminal fragment of ADAM10, consistent with earlier reports that the ADAM10 ectodomain is itself a target for sheddases. The abundance of this fragment was increased in the presence of a γ-secretase inhibitor, but was not affected by M3 receptor activation. The dynamin mutant did not affect the distribution of ADAM10 and its C-terminal fragment between raft and non-raft membrane compartments. Surface expression and limited proteolysis of ADAM10 are regulated by dynamin-dependent endocytosis, but are unaffected by activation of signaling pathways that upregulate shedding of ADAM substrates such as APP. Modulation of ADAM10 internalization could affect cellular behavior in two ways: by altering the putative signaling activity of

  5. Subtilisin QK-2: secretory expression in Lactococcus lactis and surface display onto gram-positive enhancer matrix (GEM) particles.

    Science.gov (United States)

    Mao, Ruifeng; Zhou, Kangping; Han, Zhenwei; Wang, Yefu

    2016-05-12

    Purified from the supernatant of Bacillus subtilis QK02 culture broth, Subtilisin QK-2 is a type of effective thrombolytic reagent that has great exploitable potential. However, the unbearable flavor that occurs with fermentation and the complicated methods that are required to obtain pure products limit the application of this enzyme. Lactic acid bacteria (LAB)-based delivery vehicles are promising as cheap and safe options for medicinal compounds. The secretory expression and surface display using LAB may popularize Subtilisin QK-2 more easily and conveniently with minimal adverse effects. Subtilisin QK-2 was expressed successfully in two forms using lactic acid bacteria. For the secretory expression in Lactococcus lactis, Subtilisin QK-2 was efficiently secreted into the culture using the promoter P nisA and signal peptide SPUsp. The expression levels were not different in L. lactis NZ9000 and NZ3900 without the effect of different selection markers. However, leaky expression was only detected in L. lactis NZ3900. The biological activity of this secreted Subtilisin QK-2 was enhanced by modulating the pH of medium to slightly alkaline during induction and by codon optimization of either the entire gene sequence (qk') or only the propeptide gene sequence (qkpro'). For surface display onto gram-positive enhancer matrix (GEM) particles, n LysM repeats from the C-terminal region of the major autolysin AcmA of L. lactis were fused to either the C-terminus (n = 1, 3, 5) or the N-terminus (n = 1) of the Subtilisin QK-2. These fusion proteins were secreted into the culture medium, and the QK-3LysM was able to bind to the surface of various LAB GEM particles without a loss of fibrinolytic activity. Furthermore, the binding capacity significantly increased with a higher concentration of QK-3LysM. Compared to the free-form Subtilisin QK-2, the QK-3LysM displayed on the surface of GEM particles was more stable in the simulated gastric juice. Combined with the safety and

  6. Distinct regions in the C-Terminus required for GLP-1R cell surface expression, activity and internalisation.

    Science.gov (United States)

    Thompson, Aiysha; Kanamarlapudi, Venkateswarlu

    2015-09-15

    The glucagon-like peptide-1 (GLP-1) receptor (GLP-1R), an important drug target in the treatment of type 2 diabetes, is a G-protein coupled receptor (GPCR) that mediates insulin secretion by GLP-1. The N-terminus controls GLP-1R biosynthetic trafficking to the cell surface but the C-terminus involvement in that trafficking is unknown. The aim of this study was to identify distinct regions within the C-terminal domain required for human GLP-1R (hGLP-1R) cell surface expression, activity and internalisation using a number of C-terminal deletions and site-directed mutations. The results of this study revealed that the residues 411-418 within the C-terminal domain of the hGLP-1R are critical in targeting the newly synthesised receptor to the plasma membrane. The residues 419-430 are important for cAMP producing activity of the receptor, most likely by coupling to Gαs. However, the residues 431-450 within the C-terminus are essential for agonist-induced hGLP-1R internalisation. In conclusion, these findings demonstrate the hGLP-1R has distinct regions within the C-terminal domain required for its cell surface expression, activity and agonist-induced internalisation. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  7. Genetic structure and expression of the surface glycoprotein GP82, the main adhesin of Trypanosoma cruzi metacyclic trypomastigotes.

    Science.gov (United States)

    Correa, Paulo Roberto Ceridorio; Cordero, Esteban Mauricio; Gentil, Luciana Girotto; Bayer-Santos, Ethel; da Silveira, José Franco

    2013-01-01

    T. cruzi improves the likelihood of invading or adapting to the host through its capacity to present a large repertoire of surface molecules. The metacyclic stage-specific surface glycoprotein GP82 has been implicated in host cell invasion. GP82 is encoded by multiple genes from the trans-sialidase superfamily. GP82 shows a modular organization, with some variation of N-terminal region flanking a conserved central core where the binding sites to the mammalian cell and gastric mucin are located. The function of GP82 as adhesin in host cell invasion process could expose the protein to an intense conservative and selective pressure. GP82 is a GPI-anchored surface protein, synthesized as a 70 kDa precursor devoid of N-linked sugars. GPI-minus variants accumulate in the ER indicating that GPI anchor acts as a forward transport signal for progressing along the secretory pathway as suggested for T. cruzi mucins. It has been demonstrated that the expression of GP82 is constitutive and may be regulated at post-transcriptional level, for instance, at translational level and/or mRNA stabilization. GP82 mRNAs are mobilized to polysomes and consequently translated, but only in metacyclic trypomastigotes. Analysis of transgenic parasites indicates that the mechanism regulating GP82 expression involves multiple elements in the 3'UTR.

  8. A member of the CPW-WPC protein family is expressed in and localized to the surface of developing ookinetes.

    Science.gov (United States)

    Kangwanrangsan, Niwat; Tachibana, Mayumi; Jenwithisuk, Rachaneeporn; Tsuboi, Takafumi; Riengrojpitak, Suda; Torii, Motomi; Ishino, Tomoko

    2013-04-15

    Despite the development of malaria control programs, billions of people are still at risk for this infectious disease. Recently, the idea of the transmission-blocking vaccine, which works by interrupting the infection of mosquitoes by parasites, has gained attention as a promising strategy for malaria control and eradication. To date, a limited number of surface proteins have been identified in mosquito-stage parasites and investigated as potential targets for transmission-blocking vaccines. Therefore, for the development of effective transmission-blocking strategies in epidemic areas, it is necessary to identify novel zygote/ookinete surface proteins as candidate antigens. Since the expression of many zygote/ookinete proteins is regulated post-transcriptionally, proteins that are regulated by well-known translational mediators were focused. Through in silico screening, CPW-WPC family proteins were selected as potential zygote/ookinete surface proteins. All experiments were performed in the rodent malaria parasite, Plasmodium yoelii XNL. mRNA and protein expression profiles were examined by RT-PCR and western blotting, respectively, over the course of the life cycle of the malaria parasite. Protein function was also investigated by the generation of gene-disrupted transgenic parasites. The CPW-WPC protein family, named after the unique WxC repeat domains, is highly conserved among Plasmodium species. It is revealed that CPW-WPC mRNA transcripts are transcribed in gametocytes, while CPW-WPC proteins are expressed in zygote/ookinete-stage parasites. Localization analysis reveals that one of the CPW-WPC family members, designated as PyCPW-WPC-1, is a novel zygote/ookinete stage-specific surface protein. Targeted disruption of the pycpw-wpc-1 gene caused no obvious defects during ookinete and oocyst formation, suggesting that PyCPW-WPC-1 is not essential for mosquito-stage parasite development. It is demonstrated that PyCPW-WPC-1 can be classified as a novel, post

  9. northern Tanzania

    African Journals Online (AJOL)

    Department of Zoology and Marine Biology, University of Dar es Salaam,. P O Box 35064, Dar es Salaam, Tanzania ... National Parks and neighbouring villages in northern Tanzania between 1993 and l996 (Kabigumila 1998a). Most of ..... International Congress ofChelonian Conservation. SOPTOM,. Gonfaron France. pp: ...

  10. Lactobacilli differentially modulate expression of cytokines and maturation surface markers in murine dendritic cells

    DEFF Research Database (Denmark)

    Christensen, Hanne Risager; Frøkiær, Hanne; Pestka, J.J.

    2002-01-01

    in the capacity to induce IL-12 and TNF-a production in the DC. Similar but less pronounced differences were observed among lactobacilli in the induction of IL-6 and IL-10. Although all strains up-regulated surface MHC class II and B7-2 (CD86), which is indicative of DC maturation, those lactobacilli...

  11. Optimization of the Expression of DT386-BR2 Fusion Protein in Escherichia coli using Response Surface Methodology.

    Science.gov (United States)

    Shafiee, Fatemeh; Rabbani, Mohammad; Jahanian-Najafabadi, Ali

    2017-01-01

    The aim of this study was to determine the best condition for the production of DT386-BR2 fusion protein, an immunotoxin consisting of catalytic and translocation domains of diphtheria toxin fused to BR2, a cancer specific cell penetrating peptide, for targeted eradication of cancer cells, in terms of the host, cultivation condition, and culture medium. Recombinant pET28a vector containing the codons optimized for the expression of the DT386-BR2 gene was transformed to different strains of Escherichia coli ( E. coli BL21 DE3, E. coli Rosetta DE3 and E. coli Rosetta-gami 2 DE3), followed by the induction of expression using 1 mM IPTG. Then, the strain with the highest ability to produce recombinant protein was selected and used to determine the best expression condition using response surface methodology (RSM). Finally, the best culture medium was selected. Densitometry analysis of sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the expressed fusion protein showed that E. coli Rosetta DE3 produced the highest amounts of the recombinant fusion protein when quantified by 1 mg/ml bovine serum albumin (178.07 μg/ml). Results of RSM also showed the best condition for the production of the recombinant fusion protein was induction with 1 mM IPTG for 2 h at 37°C. Finally, it was established that terrific broth could produce higher amounts of the fusion protein when compared to other culture media. In this study, we expressed the recombinant DT386-BR2 fusion protein in large amounts by optimizing the expression host, cultivation condition, and culture medium. This fusion protein will be subjected to purification and evaluation of its cytotoxic effects in future studies.

  12. Optimization of the Expression of DT386-BR2 Fusion Protein in Escherichia coli using Response Surface Methodology

    Directory of Open Access Journals (Sweden)

    Fatemeh Shafiee

    2017-01-01

    Full Text Available Background: The aim of this study was to determine the best condition for the production of DT386-BR2 fusion protein, an immunotoxin consisting of catalytic and translocation domains of diphtheria toxin fused to BR2, a cancer specific cell penetrating peptide, for targeted eradication of cancer cells, in terms of the host, cultivation condition, and culture medium. Materials and Methods: Recombinant pET28a vector containing the codons optimized for the expression of the DT386-BR2 gene was transformed to different strains of Escherichia coli (E. coli BL21 DE3, E. coli Rosetta DE3 and E. coli Rosetta-gami 2 DE3, followed by the induction of expression using 1 mM IPTG. Then, the strain with the highest ability to produce recombinant protein was selected and used to determine the best expression condition using response surface methodology (RSM. Finally, the best culture medium was selected. Results: Densitometry analysis of sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the expressed fusion protein showed that E. coli Rosetta DE3 produced the highest amounts of the recombinant fusion protein when quantified by 1 mg/ml bovine serum albumin (178.07 μg/ml. Results of RSM also showed the best condition for the production of the recombinant fusion protein was induction with 1 mM IPTG for 2 h at 37°C. Finally, it was established that terrific broth could produce higher amounts of the fusion protein when compared to other culture media. Conclusion: In this study, we expressed the recombinant DT386-BR2 fusion protein in large amounts by optimizing the expression host, cultivation condition, and culture medium. This fusion protein will be subjected to purification and evaluation of its cytotoxic effects in future studies.

  13. CXCR3 surface expression in human airway epithelial cells: cell cycle dependence and effect on cell proliferation.

    Science.gov (United States)

    Aksoy, Mark O; Yang, Yi; Ji, Rong; Reddy, P J; Shahabuddin, Syed; Litvin, Judith; Rogers, Thomas J; Kelsen, Steven G

    2006-05-01

    We recently demonstrated that human bronchial epithelial cells (HBEC) constitutively express the CXC chemokine receptor CXCR3, which when activated, induces directed cell migration. The present study in HBEC examined the relative expression of the CXCR3 splice variants CXCR3-A and -B, cell cycle dependence of CXCR3 expression, and the effects of the CXCR3 ligand, the interferon-gamma-inducible CXC chemokine I-TAC/CXCL11, on DNA synthesis and cell proliferation. Both CXCR3-A and -B mRNA, assessed by real-time RT-PCR, were expressed in normal HBEC (NHBEC) and the HBEC line 16-HBE. However, CXCR3-B mRNA was 39- and 6-fold greater than CXCR3-A mRNA in NHBEC and 16-HBE, respectively. Although most HBEC (>80%) assessed by flow cytometry and immunofluorescence microscopy contained intracellular CXCR3, only a minority (75%) were in the S + G(2)/M phases of the cell cycle. Stimulation of CXCR3 with I-TAC enhanced thymidine incorporation and cell proliferation and increased p38 and ERK1/2 phosphorylation. These data indicate that 1) human airway epithelial cells primarily express CXCR3-B mRNA, 2) surface expression of CXCR3 is largely confined to the S + G(2)/M phases of the cell cycle, and 3) activation of CXCR3 induces DNA synthesis, cell proliferation, and activation of MAPK pathways. We speculate that activation of CXCR3 exerts a mitogenic effect in HBEC, which may be important during airway mucosal injury in obstructive airway diseases such as asthma and chronic obstructive pulmonary disease.

  14. Protein phosphatase 2A isotypes regulate cell surface expression of the T cell receptor

    DEFF Research Database (Denmark)

    Lauritsen, Jens Peter Holst; Menné, C; Kastrup, J

    2001-01-01

    show that inhibition of the serine/threonine protein phosphatase PP2A family had a biphasic effect on TCR expression. Thus, low concentrations of PP2A inhibitors induced TCR down-regulation, whereas higher concentrations of PP2A inhibitors induced TCR up-regulation. The effect of PP2A inhibition...... regulatory role for PP2A in both exocytosis and endocytosis....

  15. Expression of LY6D is induced at the surface of MCF10A cells by X-ray irradiation.

    Science.gov (United States)

    Kurosawa, Maiko; Jeyasekharan, Anand D; Surmann, Eva-Maria; Hashimoto, Noboru; Venkatraman, Vignesh; Kurosawa, Gene; Furukawa, Koichi; Venkitaraman, Ashok R; Kurosawa, Yoshikazu

    2012-12-01

    In order to identify membrane proteins whose expression is induced by X-ray irradiation, we developed an antibody (Ab)-directed strategy using a phage Ab library. X-Ray-irradiated cells were screened with a phage Ab library in the presence of a large excess of polyclonal Abs prepared against membrane proteins that are commonly present at the surface of both X-ray-irradiated and nonirradiated cells. After isolation of Ab that bound only to X-ray-irradiated cells, the antigen was identified using MS. Using this approach, we found that expression of LY6D is induced in MCF10A cells by X-ray irradiation. The induction of LY6D expression is triggered through a pathway regulated by ATM, CHK2 and p53. This method is a new Ab-directed proteomic strategy for analysis of membrane proteins, and is applicable to various biological phenomena in situations in which both target molecule-expressing cells and nonexpressing cells are available. © 2012 The Authors Journal compilation © 2012 FEBS.

  16. Plasmodium falciparum variant surface antigen expression varies between isolates causing severe and nonsevere malaria and is modified by acquired immunity

    DEFF Research Database (Denmark)

    Nielsen, Morten A; Staalsoe, Trine; Kurtzhals, Jørgen

    2002-01-01

    of immunity, as disease-causing parasites appear to be those not controlled by preexisting VSA-specific Abs. In this work we report that VSA expressed by parasites from young Ghanaian children with P. falciparum malaria were commonly and strongly recognized by plasma Abs from healthy children in the same area......, and that the VSA expression pattern is modulated by immunity. This opens the possibility of developing morbidity-reducing vaccines targeting a limited subset of common and particularly virulent VSA.......In areas of endemic parasite transmission, protective immunity to Plasmodium falciparum malaria is acquired over several years with numerous disease episodes. Acquisition of Abs to parasite-encoded variant surface Ags (VSA) on the infected erythrocyte membrane is important in the development...

  17. Cooperative degradation of chitin by extracellular and cell surface-expressed chitinases from Paenibacillus sp. strain FPU-7.

    Science.gov (United States)

    Itoh, Takafumi; Hibi, Takao; Fujii, Yutaka; Sugimoto, Ikumi; Fujiwara, Akihiro; Suzuki, Fumiko; Iwasaki, Yukimoto; Kim, Jin-Kyung; Taketo, Akira; Kimoto, Hisashi

    2013-12-01

    Chitin, a major component of fungal cell walls and invertebrate cuticles, is an exceedingly abundant polysaccharide, ranking next to cellulose. Industrial demand for chitin and its degradation products as raw materials for fine chemical products is increasing. A bacterium with high chitin-decomposing activity, Paenibacillus sp. strain FPU-7, was isolated from soil by using a screening medium containing α-chitin powder. Although FPU-7 secreted several extracellular chitinases and thoroughly digested the powder, the extracellular fluid alone broke them down incompletely. Based on expression cloning and phylogenetic analysis, at least seven family 18 chitinase genes were found in the FPU-7 genome. Interestingly, the product of only one gene (chiW) was identified as possessing three S-layer homology (SLH) domains and two glycosyl hydrolase family 18 catalytic domains. Since SLH domains are known to function as anchors to the Gram-positive bacterial cell surface, ChiW was suggested to be a novel multimodular surface-expressed enzyme and to play an important role in the complete degradation of chitin. Indeed, the ChiW protein was localized on the cell surface. Each of the seven chitinase genes (chiA to chiF and chiW) was cloned and expressed in Escherichia coli cells for biochemical characterization of their products. In particular, ChiE and ChiW showed high activity for insoluble chitin. The high chitinolytic activity of strain FPU-7 and the chitinases may be useful for environmentally friendly processing of chitin in the manufacture of food and/or medicine.

  18. High-risk human papillomavirus E7 expression reduces cell-surface MHC class I molecules and increases susceptibility to natural killer cells

    DEFF Research Database (Denmark)

    Bottley, G; Watherston, O G; Hiew, Y-L

    2007-01-01

    a role for E7 in tumour immune evasion. We show that knockdown of E7 expression in HPV16- and HPV18-transformed cervical carcinoma cells by RNA interference increased expression of major histocompatibility complex (MHC) class I at the cell surface and reduced susceptibility of these cells to natural...... killer (NK) cells. Tetracycline-regulated induction of HPV16 E7 resulted in reduced expression of cell surface MHC class I molecules and increased NK cell killing. Our results suggest that, for HPV-associated malignancies, reduced MHC class I expression is the result of an active immune evasion strategy...

  19. Ultraviolet radiation (UVR) induces cell-surface Ro/SSA antigen expression by human keratinocytes in vitro: a possible mechanism for the UVR induction of cutaneous lupus lesions

    International Nuclear Information System (INIS)

    Jones, S.K.

    1992-01-01

    Antinuclear antibodies are useful markers of connective tissue disease. In this study, UVB but not UVA induced the expression of Ro/SSA antigen on keratinocyte surfaces in vitro. This expression was also found with the extractable nuclear antigens RnP and Sm, but not with single or double-stranded DNA. The expression was prevented by blocking protein synthesis, suggesting that it was an active process. The results suggest that UVB exposure may result in the expression of Ro/SSA antigen on the surfaces of basal keratinocytes in vivo. This antigen could then bind circulating antibody leading to the cutaneous lesions in neonatal and subacute cutaneous lupus erythematosus. (Author)

  20. Nanostructured implant surface effect on osteoblast gene expression and bone-to-implant contact in vivo

    Energy Technology Data Exchange (ETDEWEB)

    Mendonca, Gustavo, E-mail: gustavo_mendonca@dentistry.unc.edu [Universidade Catolica de Brasilia, Pos-Graduacao em Ciencias Genomicas e Biotecnologia, SGAN Quadra 916, Modulo B, Av. W5 Norte 70.790-160-Asa Norte Brasilia/DF (Brazil); Bone Biology and Implant Therapy Laboratory, Department of Prosthodontics, University of North Carolina at Chapel Hill, 404 Brauer Hall, CB 7450, Chapel Hill, NC 27511 (United States); Universidade Catolica de Brasilia, Curso de Odontologia, Taguatinga/DF (Brazil); Baccelli Silveira Mendonca, Daniela [Universidade Catolica de Brasilia, Pos-Graduacao em Ciencias Genomicas e Biotecnologia, SGAN Quadra 916, Modulo B, Av. W5 Norte 70.790-160-Asa Norte Brasilia/DF (Brazil) and Bone Biology and Implant Therapy Laboratory, Department of Prosthodontics, University of North Carolina at Chapel Hill, 404 Brauer Hall, CB 7450, Chapel Hill, NC 27511 (United States); Pagotto Simoes, Luis Gustavo; Araujo, Andre Luis; Leite, Edson Roberto [Departmento de Quimica, Universidade Federal de Sao Carlos-UFSCAR, Rod. Washington Luiz, 13565-905 Sao Carlos, SP (Brazil); Golin, Alexsander Luiz [Departmento de Engenharia Mecanica, Faculdade de Engenharia Mecanica, Pontificia Universidade Catolica de Curitiba, Curitiba, PR (Brazil); Aragao, Francisco J.L. [Universidade Catolica de Brasilia, Pos-Graduacao em Ciencias Genomicas e Biotecnologia, SGAN Quadra 916, Modulo B, Av. W5 Norte 70.790-160-Asa Norte Brasilia/DF (Brazil); Embrapa Recursos Geneticos e Biotecnologia, Laboratorio de Introducao e Expressao de Genes, PqEB W5 Norte, 70770-900, Brasilia, DF (Brazil); Cooper, Lyndon F., E-mail: lyndon_cooper@dentistry.unc.edu [Bone Biology and Implant Therapy Laboratory, Department of Prosthodontics, University of North Carolina at Chapel Hill, 404 Brauer Hall, CB 7450, Chapel Hill, NC 27511 (United States)

    2011-12-01

    The aim of this study was to investigate the response of nanostructured implant surfaces at the level of osteoblast differentiation and its effects in bone-to-implant contact (BIC) and removal-torque values (RTV). CpTi grade IV implants (1.6 x 4.0 mm) were machined or machined and subsequently coated with an oxide solution. The surfaces were divided into: machined (M), titania-anatase (An), titania-rutile (Ru), and zirconia (Zr). Surfaces were examined by scanning electron microscopy, atomic force microscopy, and by X-ray microanalysis. Implants were inserted in rat tibia and harvested from 0 to 21 days for measurement of Alkaline Phosphatase, Bone Sialoprotein, Osteocalcin, Osteopontin, and RUNX-2 mRNA levels by real time PCR; from 0 to 56 days for RTV; and from 0 to 56 days for BIC. The roughness parameter (Sa) was compared by one-way ANOVA followed by Tukey Test. Comparison of Torque removal values and histomorphometric measurements on implants in vivo was performed by Kruskal-Wallis test and the significance level for all statistical analyses was set at p {<=} 0.05. mRNA levels on all nanostructured surfaces were increased compared to M. At 56 days, the mean RTV in Ncm was 11.6 {+-} 2.5, 11.3 {+-} 2.4, 11.1 {+-} 3.5, 9.7 {+-} 1.4 for An, Ru, Zr, and M, respectively. Higher BIC (%) was measured for all the nanostructured surfaces versus M at 21 and 56 days (p < 0.05). Nanostructured topographic features composed of TiO{sub 2} or ZrO{sub 2} applied to machined cpTi implant promoted greater mesenchymal stem cell commitment to the osteoblast phenotype and associated increased BIC and physical association with bone. Highlights: {yields} Nanostructured surfaces using a sol-gel technique coated cpTi with TiO{sub 2} or ZrO{sub 2}. {yields} Evaluated molecular and mechanical effect of nanofeatures in vivo in rat tibiae. {yields} Nanofeatures improved the differentiation of rat MSCs into osteoblasts. {yields} Nanofeatures improved increased bone-to-implant contact and

  1. Nanostructured implant surface effect on osteoblast gene expression and bone-to-implant contact in vivo

    International Nuclear Information System (INIS)

    Mendonca, Gustavo; Baccelli Silveira Mendonca, Daniela; Pagotto Simoes, Luis Gustavo; Araujo, Andre Luis; Leite, Edson Roberto; Golin, Alexsander Luiz; Aragao, Francisco J.L.; Cooper, Lyndon F.

    2011-01-01

    The aim of this study was to investigate the response of nanostructured implant surfaces at the level of osteoblast differentiation and its effects in bone-to-implant contact (BIC) and removal-torque values (RTV). CpTi grade IV implants (1.6 x 4.0 mm) were machined or machined and subsequently coated with an oxide solution. The surfaces were divided into: machined (M), titania-anatase (An), titania-rutile (Ru), and zirconia (Zr). Surfaces were examined by scanning electron microscopy, atomic force microscopy, and by X-ray microanalysis. Implants were inserted in rat tibia and harvested from 0 to 21 days for measurement of Alkaline Phosphatase, Bone Sialoprotein, Osteocalcin, Osteopontin, and RUNX-2 mRNA levels by real time PCR; from 0 to 56 days for RTV; and from 0 to 56 days for BIC. The roughness parameter (Sa) was compared by one-way ANOVA followed by Tukey Test. Comparison of Torque removal values and histomorphometric measurements on implants in vivo was performed by Kruskal-Wallis test and the significance level for all statistical analyses was set at p ≤ 0.05. mRNA levels on all nanostructured surfaces were increased compared to M. At 56 days, the mean RTV in Ncm was 11.6 ± 2.5, 11.3 ± 2.4, 11.1 ± 3.5, 9.7 ± 1.4 for An, Ru, Zr, and M, respectively. Higher BIC (%) was measured for all the nanostructured surfaces versus M at 21 and 56 days (p 2 or ZrO 2 applied to machined cpTi implant promoted greater mesenchymal stem cell commitment to the osteoblast phenotype and associated increased BIC and physical association with bone. Highlights: → Nanostructured surfaces using a sol-gel technique coated cpTi with TiO 2 or ZrO 2 . → Evaluated molecular and mechanical effect of nanofeatures in vivo in rat tibiae. → Nanofeatures improved the differentiation of rat MSCs into osteoblasts. → Nanofeatures improved increased bone-to-implant contact and removal torque values. → TiO 2 or ZrO 2 nanofeatures improved the biological response of machined titanium.

  2. The Staphylococcus aureus Global Regulator MgrA Modulates Clumping and Virulence by Controlling Surface Protein Expression.

    Directory of Open Access Journals (Sweden)

    Heidi A Crosby

    2016-05-01

    Full Text Available Staphylococcus aureus is a human commensal and opportunistic pathogen that causes devastating infections in a wide range of locations within the body. One of the defining characteristics of S. aureus is its ability to form clumps in the presence of soluble fibrinogen, which likely has a protective benefit and facilitates adhesion to host tissue. We have previously shown that the ArlRS two-component regulatory system controls clumping, in part by repressing production of the large surface protein Ebh. In this work we show that ArlRS does not directly regulate Ebh, but instead ArlRS activates expression of the global regulator MgrA. Strains lacking mgrA fail to clump in the presence of fibrinogen, and clumping can be restored to an arlRS mutant by overexpressing either arlRS or mgrA, indicating that ArlRS and MgrA constitute a regulatory pathway. We used RNA-seq to show that MgrA represses ebh, as well as seven cell wall-associated proteins (SraP, Spa, FnbB, SasG, SasC, FmtB, and SdrD. EMSA analysis showed that MgrA directly represses expression of ebh and sraP. Clumping can be restored to an mgrA mutant by deleting the genes for Ebh, SraP and SasG, suggesting that increased expression of these proteins blocks clumping by steric hindrance. We show that mgrA mutants are less virulent in a rabbit model of endocarditis, and virulence can be partially restored by deleting the genes for the surface proteins ebh, sraP, and sasG. While mgrA mutants are unable to clump, they are known to have enhanced biofilm capacity. We demonstrate that this increase in biofilm formation is partially due to up-regulation of SasG, a surface protein known to promote intercellular interactions. These results confirm that ArlRS and MgrA constitute a regulatory cascade, and that they control expression of a number of genes important for virulence, including those for eight large surface proteins.

  3. Analysis of surface protein expression reveals the growth pattern of the gram-negative outer membrane.

    Directory of Open Access Journals (Sweden)

    Tristan S Ursell

    Full Text Available The outer membrane (OM of Gram-negative bacteria is a complex bilayer composed of proteins, phospholipids, lipoproteins, and lipopolysaccharides. Despite recent advances revealing the molecular pathways underlying protein and lipopolysaccharide incorporation into the OM, the spatial distribution and dynamic regulation of these processes remain poorly understood. Here, we used sequence-specific fluorescent labeling to map the incorporation patterns of an OM-porin protein, LamB, by labeling proteins only after epitope exposure on the cell surface. Newly synthesized LamB appeared in discrete puncta, rather than evenly distributed over the cell surface. Further growth of bacteria after labeling resulted in divergence of labeled LamB puncta, consistent with a spatial pattern of OM growth in which new, unlabeled material was also inserted in patches. At the poles, puncta remained relatively stationary through several rounds of division, a salient characteristic of the OM protein population as a whole. We propose a biophysical model of growth in which patches of new OM material are added in discrete bursts that evolve in time according to Stokes flow and are randomly distributed over the cell surface. Simulations based on this model demonstrate that our experimental observations are consistent with a bursty insertion pattern without spatial bias across the cylindrical cell surface, with approximately one burst of ≈ 10(-2 µm(2 of OM material per two minutes per µm(2. Growth by insertion of discrete patches suggests that stochasticity plays a major role in patterning and material organization in the OM.

  4. Autodisplay for the co-expression of lipase and foldase on the surface of E. coli: washing with designer bugs

    Science.gov (United States)

    2014-01-01

    Background Lipases including the lipase from Burkholderia cepacia are in a main focus in biotechnology research since many years because of their manifold possibilities for application in industrial processes. The application of Burkholderia cepacia lipase for these processes appears complicated because of the need for support by a chaperone, the lipase specific foldase. Purification and reconstitution protocols therefore interfere with an economic implementation of such enzymes in industry. Autodisplay is a convenient method to express a variety of passenger proteins on the surface of E. coli. This method makes subsequent purification steps to obtain the protein of interest unnecessary. If enzymes are used as passengers, the corresponding cells can simply be applied as whole cell biocatalysts. Furthermore, enzymes surface displayed in this manner often acquire stabilization by anchoring within the outer membrane of E. coli. Results The lipase and its chaperone foldase from B. cepacia were co-expressed on the surface of E. coli via autodisplay. The whole cell biocatalyst obtained thereby exhibited an enzymatic activity of 2.73 mU mL-1 towards the substrate p-nitrophenyl palmitate when applied in an OD578 =1. Outer membrane fractions prepared from the same culture volume showed a lipase activity of 4.01 mU mL-1. The lipase-whole cell biocatalyst as well as outer membrane preparations thereof were used in a standardized laundry test, usually adopted to determine the power of washing agents. In this test, the lipase whole cell biocatalyst and the membrane preparation derived thereof exhibited the same lipolytic activity as the purified lipase from B. cepacia and a lipase preparation which is already applied in commercial washing agents. Conclusions Co-expression of both the lipase and its chaperone foldase on the surface of E. coli yields a lipid degrading whole cell biocatalyst. Therefore the chaperone supported folding process, absolutely required for the lipolytic

  5. Bioengineered titanium surfaces affect the gene-expression and phenotypic response of osteoprogenitor cells derived from mouse calvarial bones

    Directory of Open Access Journals (Sweden)

    J Isaac

    2010-09-01

    Full Text Available This study investigated the in vitro effects of bioactive titanium surfaces on osteoblast differentiation. Three titanium substrates were tested: a commercially pure titanium (Cp Ti, an alkali- and heat-treated titanium (AH Ti, and an apatite-formed titanium (Ap Ti generated by soaking AH Ti in a simulated body fluid. Chemical evaluation of the surface reactivity was analysed at nanometre scale by X-ray photoelectron spectroscopy (XPS, and at micrometre scale by energy dispersive X-ray microanalysis (EDX. It showed that the estimated proportion of the surface covered by adsorbed serum proteins differed between the three substrates and confirmed the bioactivity of AH Ti, illustrated by surface calcium and phosphate deposition when immersed in biological fluids. Mouse calvaria osteoblasts were cultured on the substrates for 15 days with no sign of cytotoxicity. Enzyme immunoassay and Real-Time RT-PCR were used to follow osteoblast differentiation through the production of osteocalcin (OC and expression of several bone markers. At day 15, a significant up-regulation of Runx2, Osx, Dlx5, ALP, BSP, OC and DMP1 mRNA levels associated with an increase of OC production were observed on AH Ti and Ap Ti when compared to Cp Ti. These results suggest that bioengineered titanium has a great potential for dental applications in enhancing osseointegration.

  6. Northern Pintail

    Science.gov (United States)

    Clark, Robert G.; Fleskes, Joseph P.; Guyn, Karla L.; Haukos, David A.; Austin, Jane E.; Miller, Michael R.

    2014-01-01

    This medium-sized dabbling duck of slender, elegant lines and conservative plumage coloration is circumpolar in distribution and abundant in North America, with core nesting habitat in Alaska and the Prairie Pothole Region of southern Canada and the northern Great Plains. Breeders favor shallow wetlands interspersed throughout prairie grasslands or arctic tundra. An early fall migrant, the species arrives on wintering areas beginning in August, after wing molt, often forming large roosting and feeding flocks on open, shallow wetlands and flooded agricultural fields. The birds consume grains, marsh plant seeds, and aquatic invertebrates throughout fall and winter.Northern Pintails are among the earliest nesting ducks in North America, beginning shortly after ice-out in many northern areas. Individuals form new pair bonds each winter but are highly promiscuous during the nesting season, with mated and unmated males often involved in vigorous, acrobatic Pursuit Flights. Annual nest success and productivity vary with water conditions, predation, and weather. Females build nests on the ground, often far from water. Only the female incubates; her mate leaves shortly after incubation begins. Ducklings hatch together in one day, follow the female to water after a day in the nest, and fledge by July or August. Adults and ducklings consume mainly aquatic invertebrates during the breeding season.Predators and farming operations destroy many thousands of Northern Pintail nests annually; farming has also greatly reduced the amount of quality nesting cover available. Winter habitats are threatened by water shortages, agricultural development, contamination, and urbanization. Periods of extended drought in prairie nesting regions have caused dramatic population declines, usually followed by periods of recovery. Over the long term, however, the continental population of Northern Pintails has declined significantly from 6 million birds in the early 1970s to less than 3 million in

  7. Polymorphic expression in the CD8alpha chain surface receptor of African lions (Panthera leo).

    Science.gov (United States)

    Bull, Marta E; Gebhard, Douglas G; Tompkins, Wayne A F; Kennedy-Stoskopf, Suzanne

    2002-01-15

    Free-ranging African lion (Panthera leo) peripheral blood mononuclear cells (PBMC) were examined using flow cytometry and antibodies developed for use in the domestic cat to determine if phenotypic changes occurred in lion lymphocytes as a result of feline immunodeficiency virus (FIV) infection. The percentage of CD8 cells from lion peripheral blood was considerably lower than in the domestic cat. Lions with elevated levels of CD8+ cells were typically infected with FIV, similar to observations in the domestic cat. Antibodies against the alpha chain of the CD8 receptor (monoclonal antibody (mAb) 3.357) did not react consistently in all lions examined. Flow cytometric analysis determined that approximately 82 and 80% of the animals from Kruger and Hluhluwe-Umfolozi National Parks in South Africa reacted with the monoclonal antibody against the alpha chain of CD8 receptor, while only 17% of the lions in Etosha National Park in Namibia cross-reacted with the CD8alpha chain. There was no apparent correlation between FIV status and CD8alpha chain reactivity. The relative isolation of Etosha from the other two parks could explain the marked difference in CD8alpha chain expression and suggests that lions similar to other mammalian species demonstrate polymorphic expression of the CD8alpha chain (197).

  8. Comprehensive evaluation of dioxins and dioxin-like compounds in surface soils and river sediments from e-waste-processing sites in a village in northern Vietnam: Heading towards the environmentally sound management of e-waste

    Directory of Open Access Journals (Sweden)

    Go Suzuki

    2016-06-01

    Full Text Available The management of electronic waste (e-waste, which can be a source of both useful materials and toxic substances, depending on the processing method, is important for promoting material cycling. In this study, we used the dioxin-responsive chemical-activated luciferase gene expression (DR-CALUX assay combined with gas chromatography–high-resolution mass spectrometry to evaluate the levels of dioxin-like compounds in surface soils and river sediments collected in and around an e-waste-processing village in northern Vietnam. The WHO-TEQs (Toxic equivalents of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs, coplanar polychlorinated biphenyls (Co-PCBs, and polybrominated dibenzo-p-dioxins and dibenzofurans (PBDD/Fs in soils collected in January 2012 ranged from 0.29 to 310 pg/g (median 2.9 pg/g, n = 32, and the WHO-TEQs in sediments ranged from 0.96 to 58 pg/g (median 4.4 pg/g, n = 8. Dioxin-like activities (CALUX-TEQs [2,3,7,8-TCDD equivalent] in soils collected in January 2012, 2013, and 2014 ranged from <30 to 4300 pg/g (median <30 pg/g, n = 96, and the activities in sediments ranged from <30 to 4000 pg/g (median 33 pg/g, n = 24. Dioxin-like compounds accumulated in samples collected around e-waste-processing areas such as open-burning sites and e-waste-processing workshops, and the compounds may be transported from their sources to surrounding areas over the course of several years. Some of the CALUX-TEQs, but not WHO-TEQs, values were higher than the maximum acceptable WHO-TEQs promulgated by various authorities, indicating that all dioxin-like compounds should be evaluated in samples collected from e-waste-processing areas. Our findings suggest that open burning and open storage of e-waste should be prohibited and that wastewater treatment should be implemented at each workshop to reduce contamination by dioxin-like compounds from e-waste.

  9. The Brugada syndrome mutation A39V does not affect surface expression of neuronal rat Cav1.2 channels

    Directory of Open Access Journals (Sweden)

    Simms Brett A

    2012-03-01

    Full Text Available Abstract Background A loss of function of the L-type calcium channel, Cav1.2, results in a cardiac specific disease known as Brugada syndrome. Although many Brugada syndrome channelopathies reduce channel function, one point mutation in the N-terminus of Cav1.2 (A39V has been shown to elicit disease a phenotype because of a loss of surface trafficking of the channel. This lack of cell membrane expression could not be rescued by the trafficking chaperone Cavβ. Findings We report that despite the striking loss of trafficking described previously in the cardiac Cav1.2 channel, the A39V mutation while in the background of the brain isoform traffics and functions normally. We detected no differences in biophysical properties between wild type Cav1.2 and A39V-Cav1.2 in the presence of either a cardiac (Cavβ2b, or a neuronal beta subunit (Cavβ1b. In addition, the A39V-Cav1.2 mutant showed a normal Cavβ2b mediated increase in surface expression in tsA-201 cells. Conclusions The Brugada syndrome mutation A39V when introduced into rat brain Cav1.2 does not trigger the loss-of-trafficking phenotype seen in a previous study on the human heart isoform of the channel.

  10. Covalent and Oriented Surface Immobilization of Antibody Using Photoactivatable Antibody Fc-Binding Protein Expressed in Escherichia coli.

    Science.gov (United States)

    Lee, Yeolin; Jeong, Jiyun; Lee, Gabi; Moon, Jeong Hee; Lee, Myung Kyu

    2016-10-04

    Fc-specific antibody binding proteins (FcBPs) with the minimal domain of protein G are widely used for immobilization of well-oriented antibodies onto solid surfaces, but the noncovalently bound antibodies to FcBPs are unstable in sera containing large amounts of antibodies. Here we report novel photoactivatable FcBPs with photomethionine (pMet) expressed in E. coli, which induce Fc-specific photo-cross-linking with antibodies upon UV irradiation. Unfortunately, pMet did not support protein expression in the native E. coli system, and therefore we also developed an engineered methionyl tRNA synthetase (MRS5m). Coexpression of MRS5m proteins successfully induced photoactivatable FcBP overexpression in methionine-auxotroph E. coli cells. The photoactivatable FcBPs could be easily immobilized on beads and slides via their N-terminal cysteine residues and 6xHis tag. The antibodies photo-cross-linked onto the photoactivatable FcBP-beads were resistant from serum-antibody mediated dissociation and efficiently captured antigens in human sera. Furthermore, photo-cross-linked antibody arrays prepared using this system allowed sensitive detection of antigens in human sera by sandwich immunoassay. The photoactivatable FcBPs will be widely applicable for well-oriented antibody immobilization on various surfaces of microfluidic chips, glass slides, and nanobeads, which are required for development of sensitive immunosensors.

  11. Regulation of Pannexin 1 Surface Expression by Extracellular ATP: Potential Implications for Nervous System Function in Health and Disease

    Directory of Open Access Journals (Sweden)

    Leigh A. Swayne

    2017-08-01

    Full Text Available Pannexin 1 (Panx1 channels are widely recognized for their role in ATP release, and as follows, their function is closely tied to that of ATP-activated P2X7 purinergic receptors (P2X7Rs. Our recent work has shown that extracellular ATP induces clustering of Panx1 with P2X7Rs and their subsequent internalization through a non-canonical cholesterol-dependent mechanism. In other words, we have demonstrated that extracellular ATP levels can regulate the cell surface expression of Panx1. Here we discuss two situations in which we hypothesize that ATP modulation of Panx1 surface expression could be relevant for central nervous system function. The first scenario involves the development of new neurons in the ventricular zone. We propose that ATP-induced Panx1 endocytosis could play an important role in regulating the balance of cell proliferation, survival, and differentiation within this neurogenic niche in the healthy brain. The second scenario relates to the spinal cord, in which we posit that an impairment of ATP-induced Panx1 endocytosis could contribute to pathological neuroplasticity. Together, the discussion of these hypotheses serves to highlight important outstanding questions regarding the interplay between extracellular ATP, Panx1, and P2X7Rs in the nervous system in health and disease.

  12. Escherichia coli Nissle 1917 bacterial ghosts retain crucial surface properties and express chlamydial antigen: an imaging study of a delivery system for the ocular surface.

    Science.gov (United States)

    Montanaro, Jacqueline; Inic-Kanada, Aleksandra; Ladurner, Angela; Stein, Elisabeth; Belij, Sandra; Bintner, Nora; Schlacher, Simone; Schuerer, Nadine; Mayr, Ulrike Beate; Lubitz, Werner; Leisch, Nikolaus; Barisani-Asenbauer, Talin

    2015-01-01

    To target chronic inflammatory ocular surface diseases, a drug delivery platform is needed that is safe, possesses immunomodulatory properties, and can be used either for drug delivery, or as a foreign antigen carrier. A new therapeutic approach that we have previously proposed uses nonliving bacterial ghosts (BGs) as a carrier-delivery system which can be engineered to carry foreign antigens and/or be loaded with therapeutic drugs. The parent strain chosen for development of our BG delivery system is the probiotic Escherichia coli strain Nissle 1917 (EcN), whose intrinsic properties trigger the innate immune system with the flagella and fimbriae used to attach and stimulate epithelial cells. In previous studies, we have shown that EcN BGs are safe for the ocular surface route, but evidence that EcN BGs retain flagella and fimbriae after transformation, has never been visually confirmed. In this study, we used different visualization techniques to determine whether flagella and fimbriae are retained on EcN BGs engineered either for drug delivery or as a foreign antigen carrier. We have also shown by immunoelectron microscopy that EcN retains two foreign antigens after processing to become EcN BGs. Furthermore, we demonstrated that BGs derived from EcN and expressing a foreign antigen attachment to conjunctival epithelial cells in vitro without causing reduced cell viability. These results are an important step in constructing a delivery system based on a nonliving probiotic that is suitable for use in ocular surface diseases pairing immunomodulation and targeted delivery.

  13. RNA-Eluting Surfaces for the Modulation of Gene Expression as A Novel Stent Concept

    Directory of Open Access Journals (Sweden)

    Olivia Koenig

    2017-02-01

    Full Text Available Presently, a new era of drug-eluting stents is continuing to improve late adverse effects such as thrombosis after coronary stent implantation in atherosclerotic vessels. The application of gene expression–modulating stents releasing specific small interfering RNAs (siRNAs or messenger RNAs (mRNAs to the vascular wall might have the potential to improve the regeneration of the vessel wall and to inhibit adverse effects as a new promising therapeutic strategy. Different poly (lactic-co-glycolic acid (PLGA resomers for their ability as an siRNA delivery carrier against intercellular adhesion molecule (ICAM-1 with a depot effect were tested. Biodegradability, hemocompatibility, and high cell viability were found in all PLGAs. We generated PLGA coatings with incorporated siRNA that were able to transfect EA.hy926 and human vascular endothelial cells. Transfected EA.hy926 showed significant siICAM-1 knockdown. Furthermore, co-transfection of siRNA and enhanced green fluorescent protein (eGFP mRNA led to the expression of eGFP as well as to the siRNA transfection. Using our PLGA and siRNA multilayers, we reached high transfection efficiencies in EA.hy926 cells until day six and long-lasting transfection until day 20. Our results indicate that siRNA and mRNA nanoparticles incorporated in PLGA films have the potential for the modulation of gene expression after stent implantation to achieve accelerated regeneration of endothelial cells and to reduce the risk of restenosis.

  14. RANKL downregulates cell surface CXCR6 expression through JAK2/STAT3 signaling pathway during osteoclastogenesis

    Energy Technology Data Exchange (ETDEWEB)

    Li, Changhong; Zhao, Jinxia; Sun, Lin; Yao, Zhongqiang; Liu, Rui [Department of Rheumatology and Immunology, Peking University Third Hospital, Beijing 100191 (China); Huang, Jiansheng [Department of Pediatrics, Washington University School of Medicine, St. Louis, MO (United States); Liu, Xiangyuan, E-mail: liu-xiangyuan@263.net [Department of Rheumatology and Immunology, Peking University Third Hospital, Beijing 100191 (China)

    2012-12-14

    Highlights: Black-Right-Pointing-Pointer CXCR6 is down-regulated during RANKL-induced osteoclastogenesis in RAW264.7 cells. Black-Right-Pointing-Pointer CXCR6 reduction was nearly reversed by inhibition of JAK2/STAT3 signaling pathway. Black-Right-Pointing-Pointer CXCL16 alone does not positively regulate osteoclastogenesis. -- Abstract: The receptor activator of nuclear factor-{kappa}B ligand (RANKL), as a member of the tumor necrosis factor (TNF) family, plays an essential role in osteoclast differentiation and function. Chemokines and their receptors have recently been shown to play critical roles in osteoclastogenesis, however, whether CXCL16-CXCR6 plays role in RANKL-mediated osteoclastogenesis is unknown. In this study, we first reported that RANKL decreased CXCR6 in a dose-dependent manner, which may be through deactivation of Akt and STAT3 signaling induced by CXCL16. Interestingly, RANKL-mediated CXCR6 reduction may be associated to the activation of STAT3 by phosphorylation. When STAT3 activation was blocked by JAK2/STAT3 inhibitor AG490, RANKL failed to shut down CXCR6 expression during osteoclastogenesis. However, CXCL16 alone did not augment RANKL-mediated osteoclast differentiation and did not alter RANKL-receptor RANK mRNA expression. These results demonstrate that reduction of CXCL16-CXCR6 is critical in RANKL-mediated osteoclastogenesis, which is mainly through the activation of JAK2/STAT3 signaling. CXCL16-CXCR6 axis may become a novel target for the therapeutic intervention of bone resorbing diseases such as rheumatoid arthritis and osteoporosis.

  15. The mouse gamete adhesin, SED1, is expressed on the surface of acrosome-intact human sperm.

    Science.gov (United States)

    Copland, Susannah D; Murphy, Ana A; Shur, Barry D

    2009-12-01

    To determine whether SED1, a protein secreted by the mouse epididymis that coats sperm and participates in sperm adhesion to the zona pellucida, is present on human sperm and in human epididymal tissue. SED1 expression was analyzed by immunoblot and indirect immunofluorescence assays. Academic clinical and research laboratories. Human breast milk was donated. Unused semen was donated by men presenting for semen analysis or in vitro fertilization (IVF). Cadaveric epididymal tissue was obtained from the institutional body donor program. Human milk fat globule membranes and human seminal plasma proteins were analyzed by immunoblot. Human sperm and epididymis were analyzed by indirect immunofluorescence microscopy. Acrosomal status was determined by staining with fluorescein isothiocyanate-Pisum sativum agglutinin. Immunoblot and indirect immunofluorescence assays. Human SED1 is recognized by two different polyclonal anti-SED1 antisera. SED1 is localized to the plasma membrane of human sperm overlying the intact acrosome. In acrosome-reacted sperm, SED1 is localized to the equatorial segment. SED1 is expressed by the epithelium of the anterior caput epididymis. SED1 is expressed on the surface of acrosome-intact human sperm and in the anterior caput of the human epididymis, similar to that seen in mouse.

  16. Expression of variant surface antigens by Plasmodium falciparum parasites in the peripheral blood of clinically immune pregnant women indicates ongoing placental infection

    DEFF Research Database (Denmark)

    Ofori, Michael F; Staalsoe, Trine; Bam, Victoria

    2003-01-01

    Placenta-sequestered Plasmodium falciparum parasites that cause pregnancy-associated malaria (PAM) in otherwise clinically immune women express distinct variant surface antigens (VSA(PAM)) not expressed by parasites in nonpregnant individuals. We report here that parasites from the peripheral blood...

  17. CC chemokine receptor 5 cell-surface expression in relation to CC chemokine receptor 5 genotype and the clinical course of HIV-1 infection

    NARCIS (Netherlands)

    de Roda Husman, A. M.; Blaak, H.; Brouwer, M.; Schuitemaker, H.

    1999-01-01

    CCR5 cell-surface expression was studied in relation to CCR5 genotype and clinical course of HIV-1 infection. HIV-1 infected CCR5+/+ individuals had higher percentages of CCR5-expressing CD4+ T cells as compared with HIV-1-infected CCR532/+ individuals. For both genotypic groups, the percentages of

  18. How Important Is Connectivity for Surface Water Fluxes? A Generalized Expression for Flow Through Heterogeneous Landscapes

    Science.gov (United States)

    Larsen, Laurel G.; Ma, Jie; Kaplan, David

    2017-10-01

    How important is hydrologic connectivity for surface water fluxes through heterogeneous floodplains, deltas, and wetlands? While significant for management, this question remains poorly addressed. Here we adopt spatial resistance averaging, based on channel and patch configuration metrics quantifiable from aerial imagery, to produce an upscaled rate law for discharge. Our model suggests that patch coverage largely controls discharge sensitivity, with smaller effects from channel connectivity and vegetation patch fractal dimension. However, connectivity and patch configuration become increasingly important near the percolation threshold and at low water levels. These effects can establish positive feedbacks responsible for substantial flow change in evolving landscapes (14-36%, in our Everglades case study). Connectivity also interacts with other drivers; flow through poorly connected hydroscapes is less resilient to perturbations in other drivers. Finally, we found that flow through heterogeneous patches is alone sufficient to produce non-Manning flow-depth relationships commonly observed in wetlands but previously attributed to depth-varying roughness.

  19. Expression of variant surface antigens by Plasmodium falciparum parasites in the peripheral blood of clinically immune pregnant women indicates ongoing placental infection

    DEFF Research Database (Denmark)

    Ofori, Michael F; Staalsoe, Trine; Bam, Victoria

    2003-01-01

    Placenta-sequestered Plasmodium falciparum parasites that cause pregnancy-associated malaria (PAM) in otherwise clinically immune women express distinct variant surface antigens (VSA(PAM)) not expressed by parasites in nonpregnant individuals. We report here that parasites from the peripheral blood...... of clinically immune pregnant women also express VSA(PAM), making them a convenient source of VSA(PAM) expressors for PAM vaccine research....

  20. Treponema pallidum Lipoprotein TP0435 Expressed in Borrelia burgdorferi Produces Multiple Surface/Periplasmic Isoforms and mediates Adherence

    Science.gov (United States)

    Chan, Kamfai; Nasereddin, Thayer; Alter, Laura; Centurion-Lara, Arturo; Giacani, Lorenzo; Parveen, Nikhat

    2016-01-01

    The ability of Treponema pallidum, the syphilis spirochete to colonize various tissues requires the presence of surface-exposed adhesins that have been difficult to identify due to the inability to culture and genetically manipulate T. pallidum. Using a Borrelia burgdorferi-based heterologous system and gain-in-function approach, we show for the first time that a highly immunogenic lipoprotein TP0435 can be differentially processed into multiple isoforms with one variant stochastically displayed on the spirochete surface. TP0435 was previously believed to be exclusively located in T. pallidum periplasm. Furthermore, non-adherent B. burgdorferi strain expressing TP0435 acquires the ability to bind to a variety of host cells including placental cells and exhibits slow opsonophagocytosis in vitro similar to poor ex vivo phagocytosis of T. pallidum by host macrophages reported previously. This phenomenon of production of both surface and periplasmic immunogenic lipoprotein isoforms has possible implications in immune evasion of the obligate pathogen T. pallidum during infection. PMID:27161310

  1. Optimized transitory ectopic expression of promastigote surface antigen protein in Nicotiana benthamiana, a potential anti-leishmaniasis vaccine candidate.

    Science.gov (United States)

    Lacombe, Séverine; Bangratz, Martine; Brizard, Jean-Paul; Petitdidier, Elodie; Pagniez, Julie; Sérémé, Drissa; Lemesre, Jean-Loup; Brugidou, Christophe

    2018-01-01

    In recent years, plants have been shown to be an efficient alternative expression system for high-value pharmaceuticals such as vaccines. However, constitutive expression of recombinant protein remains uncertain on their level of production and biological activity. To overcome these problems, transitory expression systems have been developed. Here, a series of experiments were performed to determine the most effective conditions to enhance vaccine antigen transient accumulation in Nicotiana benthamiana leaves using the promastigote surface antigen (PSA) from the parasitic protozoan Leishmania infantum. This protein has been previously identified as the major antigen of a licensed canine anti-leishmaniasis vaccine. The classical prokaryote Escherichia coli biosystem failed in accumulating PSA. Consequently, the standard plant system based on N. benthamiana has been optimized for the production of putatively active PSA. First, the RNA silencing defense mechanism set up by the plant against PSA ectopic expression was abolished by using three viral suppressors acting at different steps of the RNA silencing pathway. Then, we demonstrated that the signal peptide at the N-terminal side of the PSA is required for its accumulation. The PSA ER signaling and retention with the PSA signal peptide and the KDEL motif, respectively were optimized to significantly increase its accumulation. Finally, we demonstrate that the production of recombinant PSA in N. benthamiana leaves allows the conservation of its immunogenic property. These approaches demonstrate that based on these optimizations, plant based systems can be used to effectively produce the biological active PSA protein. Copyright © 2017 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  2. A Bacterial Surface Display System Expressing Cleavable Capsid Proteins of Human Norovirus: A Novel System to Discover Candidate Receptors

    Directory of Open Access Journals (Sweden)

    Qian Xu

    2017-12-01

    Full Text Available Human noroviruses (HuNoVs are the dominant cause of food-borne outbreaks of acute gastroenteritis. However, fundamental researches on HuNoVs, such as identification of viral receptors have been limited by the currently immature system to culture HuNoVs and the lack of efficient small animal models. Previously, we demonstrated that the recombinant protruding domain (P domain of HuNoVs capsid proteins were successfully anchored on the surface of Escherichia coli BL21 cells after the bacteria were transformed with a plasmid expressing HuNoVs P protein fused with bacterial transmembrane anchor protein. The cell-surface-displayed P proteins could specifically recognize and bind to histo-blood group antigens (HBGAs, receptors of HuNoVs. In this study, an upgraded bacterial surface displayed system was developed as a new platform to discover candidate receptors of HuNoVs. A thrombin-susceptible “linker” sequence was added between the sequences of bacterial transmembrane anchor protein and P domain of HuNoV (GII.4 capsid protein in a plasmid that displays the functional P proteins on the surface of bacteria. In this new system, the surface-displayed HuNoV P proteins could be released by thrombin treatment. The released P proteins self-assembled into small particles, which were visualized by electron microscopy. The bacteria with the surface-displayed P proteins were incubated with pig stomach mucin which contained HBGAs. The bacteria-HuNoV P proteins-HBGAs complex could be collected by low speed centrifugation. The HuNoV P proteins-HBGAs complex was then separated from the recombinant bacterial surface by thrombin treatment. The released viral receptor was confirmed by using the monoclonal antibody against type A HBGA. It demonstrated that the new system was able to capture and easily isolate receptors of HuNoVs. This new strategy provides an alternative, easier approach for isolating unknown receptors/ligands of HuNoVs from different samples

  3. A Pilot Study on the Influence of Facial Expression on Measurements in Three-Dimensional Digital Surfaces of the Face in Infants With Cleft Lip and Palate

    DEFF Research Database (Denmark)

    Hermann, Nuno Vibe; Darvann, Tron Andre; Larsen, P.

    2016-01-01

    Objective Three-dimensional surface imaging is an increasingly popular modality for face measurements in infants with cleft lip and palate. Infants are noncompliant toward producing specific facial expressions, and selecting the appropriate moment of acquisition is challenging. The objective...... was to estimate amount and spatial distribution of deformation of the face due to facial expression in infants with cleft lip and palate and provide recommendations for an improved acquisition protocol, including a method of quality control in terms of obtaining images with true neutral expression. Material...... and Method s Three-dimensional surface images of ten 4-month-old infants with unrepaired cleft lip and palate were obtained using a 3dMDface stereophotogrammetric system. For each subject, five surface images judged as representing a neutral expression were obtained during the same photo session. Mean...

  4. β-Secretase BACE1 Promotes Surface Expression and Function of Kv3.4 at Hippocampal Mossy Fiber Synapses.

    Science.gov (United States)

    Hartmann, Stephanie; Zheng, Fang; Kyncl, Michele C; Karch, Sandra; Voelkl, Kerstin; Zott, Benedikt; D'Avanzo, Carla; Lomoio, Selene; Tesco, Giuseppina; Kim, Doo Y; Alzheimer, Christian; Huth, Tobias

    2018-04-04

    The β-secretase β-site APP-cleaving enzyme 1 (BACE1) is deemed a major culprit in Alzheimer's disease, but accumulating evidence indicates that there is more to the enzyme than driving the amyloidogenic processing of the amyloid precursor protein. For example, BACE1 has emerged as an important regulator of neuronal activity through proteolytic and, most unexpectedly, also through nonproteolytic interactions with several ion channels. Here, we identify and characterize the voltage-gated K + channel 3.4 (Kv3.4) as a new and functionally relevant interaction partner of BACE1. Kv3.4 gives rise to A-type current with fast activating and inactivating kinetics and serves to repolarize the presynaptic action potential. We found that BACE1 and Kv3.4 are highly enriched and remarkably colocalized in hippocampal mossy fibers (MFs). In BACE1 -/- mice of either sex, Kv3.4 surface expression was significantly reduced in the hippocampus and, in synaptic fractions thereof, Kv3.4 was specifically diminished, whereas protein levels of other presynaptic K + channels such as K Ca 1.1 and K Ca 2.3 remained unchanged. The apparent loss of presynaptic Kv3.4 affected the strength of excitatory transmission at the MF-CA3 synapse in hippocampal slices of BACE1 -/- mice when probed with the Kv3 channel blocker BDS-I. The effect of BACE1 on Kv3.4 expression and function should be bidirectional, as predicted from a heterologous expression system, in which BACE1 cotransfection produced a concomitant upregulation of Kv3.4 surface level and current based on a physical interaction between the two proteins. Our data show that, by targeting Kv3.4 to presynaptic sites, BACE1 endows the terminal with a powerful means to regulate the strength of transmitter release. SIGNIFICANCE STATEMENT The β-secretase β-site APP-cleaving enzyme 1 (BACE1) is infamous for its crucial role in the pathogenesis of Alzheimer's disease, but its physiological functions in the intact nervous system are only gradually

  5. Escherichia coli Nissle 1917 bacterial ghosts retain crucial surface properties and express chlamydial antigen: an imaging study of a delivery system for the ocular surface

    Directory of Open Access Journals (Sweden)

    Montanaro J

    2015-07-01

    Full Text Available Jacqueline Montanaro,1 Aleksandra Inic-Kanada,1 Angela Ladurner,1 Elisabeth Stein,1 Sandra Belij,1 Nora Bintner,1 Simone Schlacher,1 Nadine Schuerer,1 Ulrike Beate Mayr,2 Werner Lubitz,2 Nikolaus Leisch,3 Talin Barisani-Asenbauer11Laura Bassi Centres of Expertise, OCUVAC – Centre of Ocular Inflammation and Infection, Centre for Pathophysiology, Infectiology, and Immunology, Medical University of Vienna, Vienna, Austria; 2BIRD-C GmbH & Co KG, Kritzendorf, Austria; 3Department of Ecogenomics and Systems Biology, University of Vienna, Vienna, AustriaAbstract: To target chronic inflammatory ocular surface diseases, a drug delivery platform is needed that is safe, possesses immunomodulatory properties, and can be used either for drug delivery, or as a foreign antigen carrier. A new therapeutic approach that we have previously proposed uses nonliving bacterial ghosts (BGs as a carrier-delivery system which can be engineered to carry foreign antigens and/or be loaded with therapeutic drugs. The parent strain chosen for development of our BG delivery system is the probiotic Escherichia coli strain Nissle 1917 (EcN, whose intrinsic properties trigger the innate immune system with the flagella and fimbriae used to attach and stimulate epithelial cells. In previous studies, we have shown that EcN BGs are safe for the ocular surface route, but evidence that EcN BGs retain flagella and fimbriae after transformation, has never been visually confirmed. In this study, we used different visualization techniques to determine whether flagella and fimbriae are retained on EcN BGs engineered either for drug delivery or as a foreign antigen carrier. We have also shown by immunoelectron microscopy that EcN retains two foreign antigens after processing to become EcN BGs. Furthermore, we demonstrated that BGs derived from EcN and expressing a foreign antigen attachment to conjunctival epithelial cells in vitro without causing reduced cell viability. These results

  6. Uroguanylin inhibits H-ATPase activity and surface expression in renal distal tubules by a PKG-dependent pathway.

    Science.gov (United States)

    da Silva Lima, Vanessa; Crajoinas, Renato O; Carraro-Lacroix, Luciene R; Godinho, Alana N; Dias, João L G; Dariolli, Rafael; Girardi, Adriana C C; Fonteles, Manassés C; Malnic, Gerhard; Lessa, Lucília M A

    2014-09-15

    Cumulative evidence suggests that guanylin peptides play an important role on electrolyte homeostasis. We have previously reported that uroguanylin (UGN) inhibits bicarbonate reabsorption in a renal distal tubule. In the present study, we tested the hypothesis that the bicarbonaturic effect of UGN is at least in part attributable to inhibition of H(+)-ATPase-mediated hydrogen secretion in the distal nephron. By in vivo stationary microperfusion experiments, we were able to show that UGN inhibits H(+)-ATPase activity by a PKG-dependent pathway because KT5823 (PKG inhibitor) abolished the UGN effect on distal bicarbonate reabsorption and H89 (PKA inhibitor) was unable to prevent it. The in vivo results were confirmed by the in vitro experiments, where we used fluorescence microscopy to measure intracellular pH (pHi) recovery after an acid pulse with NH4Cl. By this technique, we observed that UGN and 8 bromoguanosine-cGMP (8Br-cGMP) inhibited H(+)-ATPase-dependent pHi recovery and that the UGN inhibitory effect was abolished in the presence of the PKG inhibitor. In addition, by using RT-PCR technique, we verified that Madin-Darby canine kidney (MDCK)-C11 cells express guanylate cyclase-C. Besides, UGN stimulated an increase of both cGMP content and PKG activity but was unable to increase the production of cellular cAMP content and PKA activity. Furthermore, we found that UGN reduced cell surface abundance of H+-ATPase B1 subunit in MDCK-C11 and that this effect was abolished by the PKG inhibitor. Taken together, our data suggest that UGN inhibits H(+)-ATPase activity and surface expression in renal distal cells by a cGMP/PKG-dependent pathway. Copyright © 2014 the American Physiological Society.

  7. Repression of CD24 surface protein expression by oncogenic Ras is relieved by inhibition of Raf but not MEK or PI3K

    Directory of Open Access Journals (Sweden)

    Nikitha K. Pallegar

    2015-08-01

    Full Text Available CD24 is a dynamically regulated cell surface protein. High expression of CD24 leads to progression of lung, prostrate, colon and pancreatic cancers, among others. In contrast, low expression of CD24 leads to cell proliferation and metastasis of breast cancer stem cells (BCSCs. Activating mutations in Ras are found in 30% of all human cancers. Oncogenic Ras constitutively stimulates the Raf, PI3K and Ral GDS signaling pathways, leading to cellular transformation. Previous studies have shown that expression of oncogenic Ras in breast cancer cells generates CD24- cells from CD24+ cells. However, the molecular mechanisms involved in the generation of CD24- cells were not determined. Here, we demonstrate that oncogenic Ras (RasV12 expression suppresses CD24 mRNA, protein, and promoter levels when expressed in NIH/3T3 cells. Furthermore, activation of only the Raf pathway was sufficient to downregulate CD24 mRNA and protein expression to levels similar to those seen in with RasV12 expression. In contrast, activation of the PI3K pathway downregulated mRNA expression with a partial effect on protein expression whereas activation of the RalGDS pathway only partially affected protein expression. Surprisingly, inhibition of MEK with U0126 only partially restored CD24 mRNA expression but not surface protein expression. In contrast, inhibition of Raf with sorafenib did not restore CD24 mRNA expression but significantly increased the proportion of RasV12 cells expressing CD24. Therefore, the Raf pathway is the major repressor of CD24 mRNA and protein expression, with PI3K also able to substantially inhibit CD24 expression. Moreover, these data indicate that the levels of CD24 mRNA and surface protein are independently regulated. Although inhibition of Raf by sorafenib only partially restored CD24 expression, sorafenib should still be considered as a potential therapeutic strategy to alter CD24 expression in CD24- cells, such as BCSCs.

  8. Expression, immunogenicity and variation of iron-regulated surface protein A from bovine isolates of Staphylococcus aureus.

    Science.gov (United States)

    Misra, Neha; Wines, Tyler F; Knopp, Colton L; McGuire, Mark A; Tinker, Juliette K

    2017-05-01

    Staphylococcus aureus iron-regulated surface protein A (IsdA) is a fibrinogen and fibronectin adhesin that also contributes to iron sequestration and resistance to innate immunity. IsdA is conserved in human isolates and has been investigated as a human vaccine candidate. Here we report the expression of isdA, the efficacy of anti-IsdA responses and the existence of IsdA sequence variants from bovine Staphylococcus. Clinical staphylococci were obtained from US dairy farms and assayed by PCR for the presence and expression of isdA. isdA-positive species from bovines included S. aureus, S. haemolyticus and S. chromogenes. Immunoassays on bovine milk and serum confirmed the induction and opsonophagocytic activity of anti-IsdA humoral responses. The variable region of isdA was sequenced and protein alignments predicted the presence of two main variants consistent with those from human S. aureus. Mouse antibodies against one IsdA variant reduced staphylococcal binding to fibronectin in vitro in an isotype-dependent manner. Purified IsdA variants bound distinctly to fibronectin and fibrinogen. Our findings demonstrate that variability within the C-terminus of this adhesin affects immune reactivity and binding specificity, but are consistent with the significance of IsdA in bovine disease and relevant for vaccine development. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  9. Expression, immunogenicity and variation of iron-regulated surface protein A from bovine isolates of Staphylococcus aureus

    Science.gov (United States)

    Misra, Neha; Wines, Tyler F.; Knopp, Colton L.; McGuire, Mark A.; Tinker, Juliette K.

    2017-01-01

    Abstract Staphylococcus aureus iron-regulated surface protein A (IsdA) is a fibrinogen and fibronectin adhesin that also contributes to iron sequestration and resistance to innate immunity. IsdA is conserved in human isolates and has been investigated as a human vaccine candidate. Here we report the expression of isdA, the efficacy of anti-IsdA responses and the existence of IsdA sequence variants from bovine Staphylococcus. Clinical staphylococci were obtained from US dairy farms and assayed by PCR for the presence and expression of isdA. isdA-positive species from bovines included S. aureus, S. haemolyticus and S. chromogenes. Immunoassays on bovine milk and serum confirmed the induction and opsonophagocytic activity of anti-IsdA humoral responses. The variable region of isdA was sequenced and protein alignments predicted the presence of two main variants consistent with those from human S. aureus. Mouse antibodies against one IsdA variant reduced staphylococcal binding to fibronectin in vitro in an isotype-dependent manner. Purified IsdA variants bound distinctly to fibronectin and fibrinogen. Our findings demonstrate that variability within the C-terminus of this adhesin affects immune reactivity and binding specificity, but are consistent with the significance of IsdA in bovine disease and relevant for vaccine development. PMID:28430959

  10. Cloning and expression of the receptor for human urokinase plasminogen activator, a central molecule in cell surface, plasmin dependent proteolysis

    DEFF Research Database (Denmark)

    Roldan, A.L.; Cubellis, M.V.; Masucci, M.T.

    1990-01-01

    , and therefore the capacity of cells to migrate and invade neighboring tissues. We have isolated a 1.4 kb cDNA clone coding for the entire human uPAR. An oligonucleotide synthesized on the basis of the N-terminal sequence of the purified protein was used to screen a cDNA library made from SV40 transformed human......, a size very close to that of the cloned cDNA. Expression of the uPAR cDNA in mouse cells confirms that the clone is complete and expresses a functional uPA binding protein, located on the cell surface and with properties similar to the human uPAR. Caseinolytic plaque assay, immunofluorescence analysis...... fibroblasts [Okayama and Berg (1983) Mol. Cell Biol., 3, 280-289]. The cDNA encodes a protein of 313 amino acids, preceded by a 21 residue signal peptide. A hydrophobicity plot suggests the presence of a membrane spanning domain close to the C-terminus. The cDNA hybridizes to a 1.4 kb mRNA from human cells...

  11. Expression of activated PIK3CA in ovarian surface epithelium results in hyperplasia but not tumor formation.

    Directory of Open Access Journals (Sweden)

    Shun Liang

    Full Text Available The Phosphatidylinositol 3'-kinase is a key regulator in various cancer-associated signal transduction pathways. Genetic alterations of its catalytic subunit alpha, PIK3CA, have been identified in ovarian cancer. Our in vivo data suggests that PIK3CA activation is one of the early genetic events in ovarian cancer. However, its role in malignant transformation of ovarian surface epithelium (OSE is largely unclear.Using the Müllerian inhibiting substance type II receptor (MISIIR promoter, we generated transgenic mice that expressed activated PIK3CA in the Müllerian epithelium. Overexpression of PIK3CA in OSE induced remarkable hyperplasia, but was not able to malignantly transform OSE in vivo. The consistent result was also observed in primary cultured OSEs. Although enforced expression of PIK3CA could not induce OSE anchorage-independent growth, it significantly increased anchorage-independent growth of OSE transformed by mutant K-ras.While PIK3CA activation may not be able to initiate OSE transformation, we conclude that activation of PIK3CA may be an important molecular event contributing to the maintenance of OSE transformation initiated by oncogenes such as K-ras.

  12. Alternate phase variation in expression of two major surface membrane proteins (MBA and UU376) of Ureaplasma parvum serovar 3.

    Science.gov (United States)

    Zimmerman, Carl-Ulrich R; Stiedl, Thomas; Rosengarten, Renate; Spergser, Joachim

    2009-03-01

    Ureaplasma urealyticum and Ureaplasma parvum are commensals and pathogens of the human urogenital tract and of newborn infants. There are four distinct U. parvum serovars and 10 distinct U. urealyticum serovars. Both species possess a distinct immunodominant variable surface protein, the multiple banded antigen (MBA), which shows size variability among isolates as a result of changes in the number of C-terminal repeating units. Adjacent to the MBA gene (UU375) lies UU376, which was annotated as 'Ureaplasma-specific conserved hypothetical gene'. In four different strains of U. parvum serovar 3, we demonstrated expression of UU376 by Western blot analysis and phase variation between UU376, here designated Upvmp376 (Ureaplasma phase-variable membrane protein 376), and MBA after application of selective pressure with hyperimmune antisera directed against either protein. By Southern blot analysis, we found that the switch between MBA and Upvmp376 expression is associated with a DNA inversion event in which the nonrepetitive region of the MBA gene and its putative promoter region are opposed to either the repetitive region of MBA or UU376. We propose that in U. parvum serovar 3, and presumably in all U. parvum and U. urealyticum, an inversion event at specific sites effects an alternate ON/OFF switching of the genes UU375 and UU376.

  13. Lovastatin enhances ecto-5'-nucleotidase activity and cell surface expression in endothelial cells: implication of rho-family GTPases.

    Science.gov (United States)

    Ledoux, S; Laouari, D; Essig, M; Runembert, I; Trugnan, G; Michel, J B; Friedlander, G

    2002-03-08

    Extracellular adenosine production by the GPI-anchored Ecto-5'-Nucleotidase (Ecto-5'-Nu) plays an important role in the cardiovascular system, notably in defense against hypoxia. It has been previously suggested that HMG-CoA reductase inhibitors (HRIs) could potentiate the hypoxic stimulation of Ecto-5'Nu in myocardial ischemia. In order to elucidate the mechanism of Ecto-5'-Nu stimulation by HRIs, Ecto-5'-Nu activity and expression were determined in an aortic endothelial cell line (SVAREC) incubated with lovastatin. Lovastatin enhanced Ecto-5'-Nu activity in a dose-dependent manner. This increase was not supported by de novo synthesis of the enzyme because neither the mRNA content nor the total amount of the protein were modified by lovastatin. By contrast, lovastatin enhanced cell surface expression of Ecto-5'-Nu and decreased endocytosis of Ecto-5'-Nu, as evidenced by immunostaining. This effect appeared unrelated to modifications of cholesterol content or Ecto-5'-Nu association with detergent-resistant membranes. The effect of lovastatin was reversed by mevalonate, the substrate of HMG-CoA reductase, by its isoprenoid derivative, geranyl-geranyl pyrophosphate, and by cytotoxic necrotizing factor, an activator of Rho-GTPases. Stimulation of Ecto-5'-Nu by lovastatin enhanced the inhibition of platelet aggregation induced by endothelial cells. In conclusion, lovastatin enhances Ecto-5'-Nu activity and membrane expression in endothelial cells. This effect seems independent of lowering cholesterol content but could be supported by an inhibition of Ecto-5'-Nu endocytosis through a decrease of Rho-GTPases isoprenylation.

  14. Efficacy of pyramided Bt proteins Cry1F, Cry1A.105, and cry2Ab2 expressed in Smartstax corn hybrids against lepidopteran insect pests in the northern United States.

    Science.gov (United States)

    Rule, D M; Nolting, S P; Prasifka, P L; Storer, N P; Hopkins, B W; Scherder, E F; Siebert, M W; Hendrix, W H

    2014-02-01

    Commercial field corn (Zea mays L.) hybrids transformed to express some or all of the lepidopteran insect-resistant traits present in SmartStax corn hybrids were evaluated for insecticidal efficacy against a wide range oflepidopteran corn pests common to the northern United States, during 2008 to 2011 at locations in 15 states. SmartStax hybrids contain a pyramid of two Bacillus thuringiensis (Bt) derived events for lepidopteran control: event TC1507 expressing Cry1F protein and MON 89034 expressing CrylA.105 + Cry2Ab2. These studies focused on characterization of the relative efficacy of each event when expressed alone or in combination, and compared with non-Bt hybrid. Corn hybrids containing pyramided insecticidal proteins Cry1F + Cry1A.105 + Cry2Ab2 (SmartStax) consistently showed reduced plant feeding damage by a wide range of lepidopteran larvae compared with single event and non-Bt hybrids. Corn hybrids expressing TC1507 or MON 89034 as single or pyramided events were consistently efficacious against Ostrinia nubilalis (Hübner). SmartStax hybrids had less injury from Agrotis ipsilon (Hufnagel) and Striacosta albicosta (Smith) than corn hybrids containing only event MON 89034 but were not more efficacious than single event TC1507 hybrids. Corn hybrids with event MON 89034 provided better control of Helicoverpa zea (Boddie), than event TC1507 alone. Spodoptera frugiperda (J.E. Smith) efficacy was higher for hybrids with pyramid events and single events compared with the non-Bt hybrids. The spectra of activity of events TC1507 and MON 89034 differed. The combination of TC1507 + MON 89034 provided redundant control of some pests where the spectra overlapped and thereby are expected to confer a resistance management benefit.

  15. Expression of myeloid antigens on lymphoblast surface in childhood acute lymphoblastic leukemia at diagnosis and its effect on early response to treatment: a preliminary report.

    Science.gov (United States)

    Sobol-Milejska, Grazyna; Mizia-Malarz, Agnieszka; Wos, Halina

    2013-09-01

    Immunodiagnosis of acute lymphoblastic leukemia (ALL) is based on the assessment of surface antigens. There are also cases in which both lymphoid and myeloid antigens can be found on the surface of lymphoblasts. The purpose of our research was to assess the expression of myeloid and lymphoblastic antigens in children with ALL, and to determine the impact of surface antigens on early response to treatment. 58 children [33 girls (56.9 %), 25 boys (43.2 %)] with ALL were studied. Response to treatment was assessed on days 8, 15, and 33. Univariate logistic regression analysis of the effect of myeloid antigens (MyAg) on response to treatment on days 8 and 33 revealed expression of any MyAg on lymphoblast surface as a factor associated with poor response to treatment. The multivariate logistic regression analysis of treatment response on day 33, showed that the expression of CD13 antigen on lymphoblast surface is a key factor affecting delayed remission (p = 0.03; odds ratio 0.12; 95 % CI 0.01-0.81). The expression of MyAg in childhood ALL adversely affects early response to treatment. The expression of CD13 antigen on day 33 is a key factor affecting complete remission in ALL patients.

  16. Initial attachment, subsequent cell proliferation/viability and gene expression of epithelial cells related to attachment and wound healing in response to different titanium surfaces.

    Science.gov (United States)

    An, Na; Rausch-fan, Xiaohui; Wieland, Marco; Matejka, Michael; Andrukhov, Oleh; Schedle, Andreas

    2012-12-01

    A tight seal between the epithelium and the dental implant surface is required to prevent bacterial inflammation and soft tissue recession and therefore to demonstrate a long-term success. Surface hydrophilicity was recently shown to promote osseointegration. The aim of this study was to investigate the influence of surface hydrophilicity in combination with surface topography of Ti implant surfaces on the behavior and activation/differentiation of epithelial cells using a set of in vitro experiments mimicking the implant-soft tissue contact. Hydrophobic acid-etched (A) and coarse-grit-blasted, acid-etched (SLA) surfaces and hydrophilic acid-etched (modA) and modSLA surfaces were produced. The behavior of an oral squamous cell carcinoma cell line (HSC-2) grown on all surfaces was compared through determination of cell attachment and proliferation/viability (CCK-8 and MTT assay), time-lapse microscopy of fluorescence labeled cells and determination of gene expression by real time polymerase chain reaction. Within the surfaces with similar wettability cell spreading and cell movements observed by time-lapse microscopy after one day of incubation were most pronounced on smoother (A and modA) surfaces compared to rougher (SLA and modSLA) surfaces. Within the surfaces with similar roughness the hydrophilic surfaces (modA and modSLA) showed more cell spreading and cell activity compared to the hydrophobic surfaces (A and SLA). The relative gene expressions of cytokeratin14, integrin α6, integrin β4, vinculin, transforming growth factor (TGF)-β, TGF-β1, and TGF-β3 were decreased in HSC-2 on all four types of Ti surfaces compared to control surfaces (tissue culture polystyrene; pmodA). These results suggest that surface hydrophilicity might positively influence the epithelial seal around dental implants. All tested titanium surfaces downregulate cell attachment, cell proliferation, expression of adhesion promoters, and cytokines involved in wound healing in HSC-2

  17. Enhanced cell surface expression, immunogenicity and genetic stability resulting from a spontaneous truncation of HIV Env expressed by a recombinant MVA

    International Nuclear Information System (INIS)

    Wyatt, Linda S.; Belyakov, Igor M.; Earl, Patricia L.; Berzofsky, Jay A.; Moss, Bernard

    2008-01-01

    During propagation of modified vaccinia virus Ankara (MVA) encoding HIV 89.6 Env, a few viral foci stained very prominently. Virus cloned from such foci replicated to higher titers than the parent and displayed enhanced genetic stability on passage. Sequence analysis showed a single nucleotide deletion in the 89.6 env gene of the mutant that caused a frame shift and truncation of 115 amino acids from the cytoplasmic domain. The truncated Env was more highly expressed on the cell surface, induced higher antibody responses than the full-length Env, reacted with HIV neutralizing monoclonal antibodies and mediated CD4/co-receptor-dependent fusion. Intramuscular (IM), intradermal (ID) needleless, and intrarectal (IR) catheter inoculations gave comparable serum IgG responses. However, intraoral (IO) needleless injector route gave the highest IgA in lung washings and IR gave the highest IgA and IgG responses in fecal extracts. Induction of CTL responses in the spleens of individual mice as assayed by intracellular cytokine staining was similar with both the full-length and truncated Env constructs. Induction of acute and memory CTL in the spleens of mice immunized with the truncated Env construct by ID, IO, and IR routes was comparable and higher than by the IM route, but only the IR route induced CTL in the gut-associated lymphoid tissue. Thus, truncation of Env enhanced genetic stability as well as serum and mucosal antibody responses, suggesting the desirability of a similar modification in MVA-based candidate HIV vaccines

  18. Adverbial descriptions in Northern Sotho | Pretorius | South African ...

    African Journals Online (AJOL)

    Owing to a virtual lack of a 'basic adverbs' word class in Northern Sotho, the objective of this article is to investigate some language strategies which are exploited to express adverbial meanings in this language. In popular Northern Sotho grammar books, it is clearly stated that adverbs in Northern Sotho are usually derived ...

  19. Restricted Cell Surface Expression of Receptor Tyrosine Kinase ROR1 in Pediatric B-Lineage Acute Lymphoblastic Leukemia Suggests Targetability with Therapeutic Monoclonal Antibodies

    Science.gov (United States)

    Dave, Hema; Anver, Miriam R.; Butcher, Donna O.; Brown, Patrick; Khan, Javed; Wayne, Alan S.; Baskar, Sivasubramanian; Rader, Christoph

    2012-01-01

    Background Despite high cure rates for pediatric B-lineage acute lymphoblastic leukemia (B-ALL), short-term and long-term toxicities and chemoresistance are shortcomings of standard chemotherapy. Immunotherapy and chemoimmunotherapy based on monoclonal antibodies (mAbs) that target cell surface antigens with restricted expression in pediatric B-ALL may offer the potential to reduce toxicities and prevent or overcome chemoresistance. The receptor tyrosine kinase ROR1 has emerged as a candidate for mAb targeting in select B-cell malignancies. Methodology and Principal Findings Using flow cytometry, Western blotting, immunohistochemistry, and confocal immunofluorescence microscopy, we analyzed the cell surface expression of ROR1 across major pediatric ALL subtypes represented by 14 cell lines and 56 primary blasts at diagnosis or relapse as well as in normal adult and pediatric tissues. Cell surface ROR1 expression was found in 45% of pediatric ALL patients, all of which were B-ALL, and was not limited to any particular genotype. All cell lines and primary blasts with E2A-PBX1 translocation and a portion of patients with other high risk genotypes, such as MLL rearrangement, expressed cell surface ROR1. Importantly, cell surface ROR1 expression was found in many of the pediatric B-ALL patients with multiply relapsed and refractory disease and normal karyotype or low risk cytogenetics, such as hyperdiploidy. Notably, cell surface ROR1 was virtually absent in normal adult and pediatric tissues. Conclusions and Significance Collectively, this study suggests that ROR1 merits preclinical and clinical investigations as a novel target for mAb-based therapies in pediatric B-ALL. We propose cell surface expression of ROR1 detected by flow cytometry as primary inclusion criterion for pediatric B-ALL patients in future clinical trials of ROR1-targeted therapies. PMID:23285131

  20. Restricted cell surface expression of receptor tyrosine kinase ROR1 in pediatric B-lineage acute lymphoblastic leukemia suggests targetability with therapeutic monoclonal antibodies.

    Directory of Open Access Journals (Sweden)

    Hema Dave

    Full Text Available Despite high cure rates for pediatric B-lineage acute lymphoblastic leukemia (B-ALL, short-term and long-term toxicities and chemoresistance are shortcomings of standard chemotherapy. Immunotherapy and chemoimmunotherapy based on monoclonal antibodies (mAbs that target cell surface antigens with restricted expression in pediatric B-ALL may offer the potential to reduce toxicities and prevent or overcome chemoresistance. The receptor tyrosine kinase ROR1 has emerged as a candidate for mAb targeting in select B-cell malignancies.Using flow cytometry, Western blotting, immunohistochemistry, and confocal immunofluorescence microscopy, we analyzed the cell surface expression of ROR1 across major pediatric ALL subtypes represented by 14 cell lines and 56 primary blasts at diagnosis or relapse as well as in normal adult and pediatric tissues. Cell surface ROR1 expression was found in 45% of pediatric ALL patients, all of which were B-ALL, and was not limited to any particular genotype. All cell lines and primary blasts with E2A-PBX1 translocation and a portion of patients with other high risk genotypes, such as MLL rearrangement, expressed cell surface ROR1. Importantly, cell surface ROR1 expression was found in many of the pediatric B-ALL patients with multiply relapsed and refractory disease and normal karyotype or low risk cytogenetics, such as hyperdiploidy. Notably, cell surface ROR1 was virtually absent in normal adult and pediatric tissues.Collectively, this study suggests that ROR1 merits preclinical and clinical investigations as a novel target for mAb-based therapies in pediatric B-ALL. We propose cell surface expression of ROR1 detected by flow cytometry as primary inclusion criterion for pediatric B-ALL patients in future clinical trials of ROR1-targeted therapies.

  1. Monitoring Changes in the Abundance of Endogenously Expressed ATP-Sensitive Potassium (KATP) Channels in the Plasma Membrane Using Surface Biotinylation.

    Science.gov (United States)

    Ruan, Jing-Syuna; Chen, Pei-Chun

    2018-01-01

    The conductance of K ATP channel activity can be regulated by gating and/or surface expression. Gating analysis is usually performed by electrophysiological recording. Analysis of surface K ATP channel expression levels requires cell fractionation, protein separation, and quantification. Cell fractionation involves time-consuming high-speed centrifugation steps and skilled techniques for taking out specific layers. Moreover, contamination of intracellular membranes can confound results. Although commercial kits have been developed to make it easier for scientists, qualities of these kits vary which can give rise to variable results. Detection of membrane proteins using surface biotinylation technique consists of labeling cell surface proteins with a biotin reagent before lysing the cells, and isolating these tagged proteins by NeutrAvidin pulldown. Then, the samples are subjected to SDS-PAGE separation, transferred to PVDF membranes, and probed with specific antibodies. Quantification of cell surface expression is accomplished by densitometric measurement of the bands corresponding to the protein of interest and subsequent normalization by a membrane protein (as control). This alternative method for detecting expression of surface protein is relatively easy in steps and more economical in comparison to other methods such as cell fractionation.

  2. Gene expression profiling supports the hypothesis that human ovarian surface epithelia are multipotent and capable of serving as ovarian cancer initiating cells

    Directory of Open Access Journals (Sweden)

    Matyunina Lilya V

    2009-12-01

    Full Text Available Abstract Background Accumulating evidence suggests that somatic stem cells undergo mutagenic transformation into cancer initiating cells. The serous subtype of ovarian adenocarcinoma in humans has been hypothesized to arise from at least two possible classes of progenitor cells: the ovarian surface epithelia (OSE and/or an as yet undefined class of progenitor cells residing in the distal end of the fallopian tube. Methods Comparative gene expression profiling analyses were carried out on OSE removed from the surface of normal human ovaries and ovarian cancer epithelial cells (CEPI isolated by laser capture micro-dissection (LCM from human serous papillary ovarian adenocarcinomas. The results of the gene expression analyses were randomly confirmed in paraffin embedded tissues from ovarian adenocarcinoma of serous subtype and non-neoplastic ovarian tissues using immunohistochemistry. Differentially expressed genes were analyzed using gene ontology, molecular pathway, and gene set enrichment analysis algorithms. Results Consistent with multipotent capacity, genes in pathways previously associated with adult stem cell maintenance are highly expressed in ovarian surface epithelia and are not expressed or expressed at very low levels in serous ovarian adenocarcinoma. Among the over 2000 genes that are significantly differentially expressed, a number of pathways and novel pathway interactions are identified that may contribute to ovarian adenocarcinoma development. Conclusions Our results are consistent with the hypothesis that human ovarian surface epithelia are multipotent and capable of serving as the origin of ovarian adenocarcinoma. While our findings do not rule out the possibility that ovarian cancers may also arise from other sources, they are inconsistent with claims that ovarian surface epithelia cannot serve as the origin of ovarian cancer initiating cells.

  3. Construction and expression of hepatitis B surface antigen escape variants within the "a" determinant by site directed mutagenesis.

    Science.gov (United States)

    Golsaz Shirazi, Forough; Amiri, Mohammad Mehdi; Mohammadi, Hamed; Bayat, Ali Ahmad; Roohi, Azam; Khoshnoodi, Jalal; Zarnani, Amir Hassan; Jeddi-Tehrani, Mahmood; Kardar, Gholam Ali; Shokri, Fazel

    2013-09-01

    The antibody response to hepatitis B surface antigen (HBsAg) controls hepatitis B virus infection. The "a" determinant of HBsAg is the most important target for protective antibody response, diagnosis and immunoprophylaxis. Mutations in this area may induce immune escape mutants and affect the performance of HBsAg assays. To construct clinically relevant recombinant mutant forms of HBsAg and assessment of their reactivity with anti-HBs monoclonal antibodies (MAbs). Wild type (wt) and mutant (mt) HBsAg genes were constructed by site directed mutagenesis and SEOing PCR. The amplified genes were inserted into pCMV6-neo plasmid and transfected in CHO cell line. The expression of wt- and mtHBsAg was assessed by commercial ELISA assays and stable cells were established and cloned by limiting dilution. The recombinant mutants were further characterized using a panel of anti-HBs monoclonal antibodies (MAbs) and the pattern of their reactivity was assessed by ELISA. Ten HBsAg mutants having single mutation within the "a" determinant including P120E, T123N, Q129H, M133L, K141E, P142S, D144A, G145R, N146S and C147S together with a wt form were successfully constructed and expressed in CHO cells. Reactivity of anti-HBs MAbs with mtHBsAgs displayed different patterns. The effect of mutations on antibody binding differed depending on the amino acid involved and its location within the ''a'' determinant. Mutation at amino acids 123 and 145 resulted in either complete loss or significant reduction of binding to all anti-HBs MAbs. Our panel of mtHBsAgs is a valuable tool for assessment of the antibody response to HBV escape mutants and may have substantial implications in HBV immunological diagnostics.

  4. High-Level Systemic Expression of Conserved Influenza Epitope in Plants on the Surface of Rod-Shaped Chimeric Particles

    Directory of Open Access Journals (Sweden)

    Natalia V. Petukhova

    2014-04-01

    Full Text Available Recombinant viruses based on the cDNA copy of the tobacco mosaic virus (TMV genome carrying different versions of the conserved M2e epitope from influenza virus A cloned into the coat protein (CP gene were obtained and partially characterized by our group previously; cysteines in the human consensus M2e sequence were changed to serine residues. This work intends to show some biological properties of these viruses following plant infections. Agroinfiltration experiments on Nicotiana benthamiana confirmed the efficient systemic expression of M2e peptides, and two point amino acid substitutions in recombinant CPs significantly influenced the symptoms and development of viral infections. Joint expression of RNA interference suppressor protein p19 from tomato bushy stunt virus (TBSV did not affect the accumulation of CP-M2e-ser recombinant protein in non-inoculated leaves. RT-PCR analysis of RNA isolated from either infected leaves or purified TMV-M2e particles proved the genetic stability of TMV‑based viral vectors. Immunoelectron microscopy of crude plant extracts demonstrated that foreign epitopes are located on the surface of chimeric virions. The rod‑shaped geometry of plant-produced M2e epitopes is different from the icosahedral or helical filamentous arrangement of M2e antigens on the carrier virus-like particles (VLP described earlier. Thereby, we created a simple and efficient system that employs agrobacteria and plant viral vectors in order to produce a candidate broad-spectrum flu vaccine.

  5. Na+/H+ exchange regulatory factor 1 is required for ROMK1 K+ channel expression in the surface membrane of cultured M-1 cortical collecting duct cells.

    Science.gov (United States)

    Suzuki, Takashi; Nakamura, Kazuyoshi; Mayanagi, Taira; Sobue, Kenji; Kubokawa, Manabu

    2017-07-22

    The ROMK1 K + channel, a member of the ROMK channel family, is the major candidate for the K + secretion pathway in the renal cortical collecting duct (CCD). ROMK1 possesses a PDZ domain-binding motif at its C-terminus that is considered a modulator of ROMK1 expression via interaction with Na + /H + exchange regulatory factor (NHERF) 1 and NHERF2 scaffold protein. Although NHERF1 is a potential binding partner of the ROMK1 K + channel, the interaction between NHERF1 and K + channel activity remains unclear. Therefore, in this study, we knocked down NHERF1 in cultured M-1 cells derived from mouse CCD and investigated the surface expression and K + channel current in these cells after exogenous transfection with EGFP-ROMK1. NHERF1 knockdown resulted in reduced surface expression of ROMK1 as indicated by a cell biotinylation assay. Using the patch-clamp technique, we further found that the number of active channels per patched membrane and the Ba 2+ -sensitive whole-cell K + current were decreased in the knockdown cells, suggesting that reduced K + current was accompanied by decreased surface expression of ROMK1 in the NHERF1 knockdown cells. Our results provide evidence that NHERF1 mediates K + current activity through acceleration of the surface expression of ROMK1 K + channels in M-1 cells. Copyright © 2017 Elsevier Inc. All rights reserved.

  6. An investigation of interactions between hypocretin/orexin signaling and glutamate receptor surface expression in the rat nucleus accumbens under basal conditions and after cocaine exposure.

    Science.gov (United States)

    Plaza-Zabala, Ainhoa; Li, Xuan; Milovanovic, Mike; Loweth, Jessica A; Maldonado, Rafael; Berrendero, Fernando; Wolf, Marina E

    2013-12-17

    Hypocretin peptides are critical for the effects of cocaine on excitatory synaptic strength in the ventral tegmental area (VTA). However, little is known about their role in cocaine-induced synaptic plasticity in the nucleus accumbens (NAc). First, we tested whether hypocretin-1 by itself could acutely modulate glutamate receptor surface expression in the NAc, given that hypocretin-1 in the VTA reproduces cocaine's effects on glutamate transmission. We found no effect of hypocretin-1 infusion on AMPA or NMDA receptor surface expression in the NAc, measured by biotinylation, either 30 min or 3h after the infusion. Second, we were interested in whether changes in hypocretin receptor-2 (Hcrtr-2) expression contribute to cocaine-induced plasticity in the NAc. As a first step towards addressing this question, Hcrtr-2 surface expression was compared in the NAc after withdrawal from extended-access self-administration of saline (control) versus cocaine. We found that surface Hcrtr-2 levels remain unchanged following 14, 25 or 48 days of withdrawal from cocaine, a time period in which high conductance GluA2-lacking AMPA receptors progressively emerge in the NAc. Overall, our results fail to support a role for hypocretins in acute modulation of glutamate receptor levels in the NAc or a role for altered Hcrtr-2 expression in withdrawal-dependent synaptic adaptations in the NAc following cocaine self-administration. Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.

  7. Associations of IGFBP3 Gene Polymorphism and Gene Expression with the Risk of Osteonecrosis of the Femoral Head in a Han Population in Northern China.

    Science.gov (United States)

    Song, Yang; Du, Zhenwu; Yang, Qiwei; Ren, Ming; Sui, Yujie; Wang, Qingyu; Wang, Ao; Zhao, Haiyue; Wang, Jincheng; Zhang, Guizhen

    2016-12-01

    The critical roles of IGFBP3 in regulating the osteogenic and adipogenic differentiation of bone marrow mesenchymal stem cells strongly indicate its potential effects on the pathogenesis of osteonecrosis of the femoral head (ONFH). In this study, we investigated the association of IGFBP3 gene polymorphism and its protein expression with the development of ONFH to further explore its molecular pathogenesis. Ligase detection reactions and enzyme-linked immunosorbent assay were used to detect the polymorphisms of rs2453839[C/T] and rs3110697[A/G] and the serum protein expression of IGFBP3 gene in 182 cases and 179 controls, respectively. The serum lipids level was also measured by automatic biochemistry analyzer. The results revealed that the recessive model of rs3110697 and the dominant model of rs2453839 were significantly associated with the increased risk of ONFH (p = 0.048, p = 0.047, respectively). The genotypes of rs2453839 were also significantly related to the clinical stages of ONFH (p = 0.017). More importantly, the serum protein expression of IGFBP3 and insulin-like growth factor 1 (IGF1) in the ONFH group were statistically increased compared with the control group (p = 0.044, p = 0.007). The serum triglyceride and low-density lipoprotein cholesterol level in the ONFH group were significantly higher than the control group (p = 0.01, p = 0.005, respectively), but the serum high-density lipoprotein cholesterol level of the ONFH group was dramatically lower than the control group (p = 0.0001). Our results showed that both the gene polymorphisms of IGFBP3 and the abnormal protein expression of serum IGFBP3 and IGF1 closely associated with the development of ONFH.

  8. Significant Associations of SOX9 Gene Polymorphism and Gene Expression with the Risk of Osteonecrosis of the Femoral Head in a Han Population in Northern China

    Directory of Open Access Journals (Sweden)

    Yang Song

    2016-01-01

    Full Text Available Sex determining region Y-box 9 (SOX9 is a key transcription factor involved in cartilage formation during the embryonic development stage and cartilage growth and repair after birth. To explore the roles of polymorphism and expression of the SOX9 gene in the development of osteonecrosis of the femoral head (ONFH, we analyzed the polymorphism of rs12601701 [A/G] and rs1042667 [A/C] and the serum protein expression of the SOX9 gene in 182 patients with ONFH and 179 healthy control subjects. Results revealed that the A-A haplotype of SOX9 gene as well as the GG and AA genotypes of rs12601701 was significantly associated with increased ONFH risk (P=0.038 and the risk of bilateral hip lesions of ONFH (P=0.009, respectively. The C-A, A-A, and A-G haplotypes were also statistically associated with the decreased and increased risk of bilateral hip lesions of ONFH (P=0.03, P=0.048, and P=0.013, respectively, while the A-A haplotype closely related to the clinical stages of ONFH (P=0.041. More importantly, the serum SOX9 protein expression of the ONFH group was greatly decreased compared to control group (P=0.0001. Our results first showed that the gene polymorphism and gene expression of SOX9 were significantly associated with the risk and clinical phenotypes of ONFH and also indicate that the SOX9 gene may play a key role in the development of ONFH.

  9. Looped host defense peptide CLP-19 binds to microtubules and inhibits surface expression of TLR4 on mouse macrophages.

    Science.gov (United States)

    Li, Di; Liu, Yao; Yang, Ya; Chen, Jian-hong; Yang, Jie; Zou, Lin-yun; Tian, Zhi-qiang; Lv, Jun; Xia, Pei-yuan

    2013-06-15

    The looped host defense peptide CLP-19 is derived from a highly functional core region of the Limulus anti-LPS factor and exerts robust anti-LPS activity by directly interacting with LPS in the extracellular space. We previously showed that prophylactic administration of CLP-19 even 20 h prior to LPS challenge might significantly increase the survival rate in a lethal endotoxin shock mouse model. Such an effect may be associated with immune regulation of CLP-19. To investigate the underlying mechanisms, peptide affinity chromatography, immunofluorescence, and Western blotting procedures were used to identify α- and β-tubulin as direct and specific binding partners of CLP-19 in the mouse macrophage cell line RAW 264.7. Bioinformatic analysis using the AutoDock Vina molecular docking and PyMOL molecular graphics system predicted that CLP-19 would bind to the functional residues of both α- and β-tubulin and would be located within the groove of microtubules. Tubulin polymerization assay revealed that CLP-19 might induce polymerization of microtubules and prevent depolymerization. The immunoregulatory effect of CLP-19 involving microtubules was investigated by flow cytometry, immunofluorescence, and Western blotting, which showed that CLP-19 prophylactic treatment of RAW 264.7 cells significantly inhibited LPS-induced surface expression of TLR4. Taken together, these results suggest that CLP-19 binding to microtubules disrupts the dynamic equilibrium of microtubules, reducing the efficacy of microtubule-dependent vesicular transport that would otherwise translocate TLR4 from the endoplasmic reticulum to the cell surface.

  10. Differential down-modulation of HLA-G and HLA-A2 or -A3 cell surface expression following human cytomegalovirus infection.

    Science.gov (United States)

    Pizzato, Nathalie; Garmy-Susini, Barbara; Le Bouteiller, Philippe; Lenfant, Françoise

    2004-06-01

    During pregnancy, the non-classical major histocompatibility complex (MHC) class I HLA-G molecule is specifically expressed in trophoblast cells at the materno-fetal interface and may exert a local control of the immune response against viral infections. Human cytomegalovirus (HCMV) infection, which is the major cause of congenital defects, encodes multiple glycoproteins (US2, US3, US6, US10 and US11) that interrupt the MHC class I pathway of antigen presentation. The effect of some of these unique short (US) proteins on HLA-G expression has been previously studied, but little is known about the modulation of HLA-G cell surface expression during the course of HCMV infection which ensures expression of all of these US proteins. Using flow cytometry analysis, HLA-G cell surface expression was evaluated in HCMV-infected U373-HLA-G transfectant cells and compared with the modulation of the endogenous classical HLA-A2 molecules. The results indicated that HCMV infection down-modulated HLA-G cell surface expression, but later after infection and to a lesser extent than HLA-A2. Using various HLA-G/HLA-A2 chimeras, we showed that the unique structure of HLA-G cytoplasmic tail was partly involved in the resistance of HLA-G to viral down-modulation. Such limited down-modulation of HLA-G may have functional consequences in term of innate immunity against congenital HCMV infection.

  11. Using a coupled groundwater/surface-water model to predict climate-change impacts to lakes in the Trout Lake Watershed, northern Wisconsin

    Science.gov (United States)

    Hunt, Randall; Walker, John F.; Markstrom, Steven L.; Hay, Lauren E.; Doherty, John; Webb, Richard M.T.; Semmens, Darius J.

    2009-01-01

    A major focus of the U.S. Geological Survey’s Trout Lake Water, Energy, and Biogeochemical Budgets (WEBB) project is the development of a watershed model to allow predictions of hydrologic response to future conditions including land-use and climate change. The coupled groundwater/surface-water model GSFLOW was chosen for this purpose because it could easily incorporate an existing groundwater flow model and it provides for simulation of surface-water processes.

  12. Edmonston Measles Virus Prevents Increased Cell Surface Expression of Peptide-Loaded Major Histocompatibility Complex Class II Proteins in Human Peripheral Monocytes

    OpenAIRE

    Yilla, Mamadi; Hickman, Carole; McGrew, Marcia; Meade, Elizabeth; Bellini, William J.

    2003-01-01

    Gamma interferon (IFN-γ) induces expression of the gene products of the major histocompatibility complex (MHC), whereas IFN-α/β can interfere with or suppress class II protein expression. In separate studies, measles virus (MV) was reported to induce IFN-α/β and to up-regulate MHC class II proteins. In an attempt to resolve this paradox, we examined the surface expression of MHC class I and class II proteins in MV-infected peripheral monocytes in the presence and absence of IFN-α/β. Infection...

  13. Lithospheric structure of the Northern Ordos and adjacent regions from surface wave tomography: implications to the tectonics of the North China Craton

    Science.gov (United States)

    LI, S.; Guo, Z.; Chen, Y. J.

    2017-12-01

    We present a high-resolution upper mantle S velocity model of the northern Ordos block using ambient noise tomography and two-plane-wave tomography between 8 and 143 s. The Ordos block, regarded as the nuclei of the Archean craton of North China Craton, is underlain by high velocity down to 200 km, indicating the preservation of cratonic root at the interior. However, thick lithospheric keel (≥ 200 km) is not observed outside the Ordos, suggesting craton reworking around the Ordos. The most important findings is the prominent low velocity shown beneath the Datong volcano that migrates westward with depth. At 200 km depth, the low velocity locates almost 500 km west to the leading edge of the flat-lying Pacific slab in the mantle transition zone. This observation is in conflict with the previous interpretation that the Datong volcano is fed by the deep upwelling related to the subduction of the Pacific plate. The westward tilted low velocity beneath the Datong volcano, however, is in agreement with the predominant NW-SE trending alignment of fast direction revealed by SKS splitting in this area, suggesting the Datong volcano is likely due to the asthenospheric mantle flow from west. Two possible scenarios could be related to this mantle process. First, the low velocity beneath the Datong volcano may link to the large-scale, deep-rooted mantle upwelling beneath the Mongolia, northwest to the Datong volcano at deeper depth revealed by Zhang et al. (2016). We postulate that when the raising mantle materials reaches the shallow depth, it would be forced bent by the thick lithosphere beneath the Gobi in Mongolia and flow southeastward to Datong volcano. Second, it is also worth noting that the low velocity beneath the Datong volcano connects to the low velocity zone (LVZ) beneath the Ordos block below 200km, which further links the LVZ beneath the northeastern Tibet to the west. Therefore, the Datong volcano could be fed by the mantle flow from northeastern Tibet. The

  14. Induction of Boosted Immune Response in Mice by Leptospiral Surface Proteins Expressed in Fusion with DnaK

    Directory of Open Access Journals (Sweden)

    Marina V. Atzingen

    2014-01-01

    Full Text Available Leptospirosis is an important global disease of human and veterinary concern. Caused by pathogenic Leptospira, the illness was recently classified as an emerging infectious disease. Currently available veterinarian vaccines do not induce long-term protection against infection and do not provide cross-protective immunity. Several studies have suggested the use of DnaK as an antigen in vaccine formulation, due to an exceptional degree of immunogenicity. We focused on four surface proteins: rLIC10368 (Lsa21, rLIC10494, rLIC12690 (Lp95, and rLIC12730, previously shown to be involved in host-pathogen interactions. Our goal was to evaluate the immunogenicity of the proteins genetically fused with DnaK in animal model. The chosen genes were amplified by PCR methodology and cloned into pAE, an E. coli vector. The recombinant proteins were expressed alone or in fusion with DnaK at the N-terminus. Our results demonstrate that leptospiral proteins fused with DnaK have elicited an enhanced immune response in mice when compared to the effect promoted by the individual proteins. The boosted immune effect was demonstrated by the production of total IgG, lymphocyte proliferation, and significant amounts of IL-10 in supernatant of splenocyte cell cultures. We believe that this approach could be employed in vaccines to enhance presentation of antigens of Leptospira to professional immune cells.

  15. Expression and Evaluation of Recombinant Plasmodium knowlesi Merozoite Surface Protein-3 (MSP-3 for Detection of Human Malaria.

    Directory of Open Access Journals (Sweden)

    Jeremy Ryan De Silva

    Full Text Available Malaria remains a major health threat in many parts of the globe and causes high mortality and morbidity with 214 million cases of malaria occurring globally in 2015. Recent studies have outlined potential diagnostic markers and vaccine candidates one of which is the merozoite surface protein (MSP-3. In this study, novel recombinant Plasmodium knowlesi MSP-3 was cloned, expressed and purified in an Escherichia coli system. Subsequently, the recombinant protein was evaluated for its sensitivity and specificity. The recombinant pkMSP-3 protein reacted with sera from patients with P. knowlesi infection in both Western blot (61% and ELISA (100%. Specificity-wise, pkMSP-3 did not react with healthy donor sera in either assay and only reacted with a few non-malarial parasitic patient sera in the ELISA assay (3 of 49. In conclusion, sensitivity and specificity of pkMSP-3 was found to be high in the ELISA and Western Blot assay and thus utilising both assays in tandem would provide the best sero-diagnostic result for P. knowlesi infection.

  16. Cigarette Smoke Suppresses the Surface Expression of c-kit and FcεRI on Mast Cells

    Directory of Open Access Journals (Sweden)

    M. E. Givi

    2013-01-01

    Full Text Available Chronic obstructive pulmonary disease (COPD is a multicomponent disease characterized by emphysema and/or chronic bronchitis. COPD is mostly associated with cigarette smoking. Cigarette smoke contains over 4,700 chemical compounds, including free radicals and LPS (a Toll-Like Receptor 4 agonist at concentrations which may contribute to the pathogenesis of diseases like COPD. We have previously shown that short-term exposure to cigarette smoke medium (CSM can stimulate several inflammatory cells via TLR4 and that CSM reduces the degranulation of bone-marrow-derived mast cells (BMMCs. In the current study, the effect of CSM on mast cells maturation and function was investigated. Coculturing of BMMC with CSM during the development of bone marrow progenitor cells suppressed the granularity and the surface expression of c-kit and FcεRI receptors. Stimulation with IgE/antigen resulted in decreased degranulation and release of Th1 and Th2 cytokines. The effects of CSM exposure could not be mimicked by the addition of LPS to the culture medium. In conclusion, this study shows that CSM may affect mast cell development and subsequent response to allergic activation in a TLR4-independent manner.

  17. Expression of the glycoprotein of viral haemorrhagic septicaemia virus (VHSV) on the surface of the fish cell line RTG-P1 induces type 1 interferon expression in neighbouring cells

    DEFF Research Database (Denmark)

    Acosta, F.; Collet, B.; Lorenzen, Niels

    2006-01-01

    In the present study using a luciferase/Mx promoter reporter system, it was shown that the rainbow trout gonad cell line (RTG-P1), a fibroblastic cell line, produces IFN when transfected with a plasmid encoding the glycoprotein of VHSV but not with plasmid vector alone. Only a small percentage...... of the cells expressed the G protein on the surface membrane as indicated by immunostaining of transfected cells. When transfection was performed in the presence of monoclonal antibodies (Mab) to the glycoprotein, the production of interferon mRNA transcripts was reduced by over 50%. This indicates...... that the surface expression of G protein was the major mechanism of interferon induction and that most of the interferon was being expressed by cells neighbouring the transfected cells. Crown...

  18. Hydrogeology, groundwater levels, and generalized potentiometric-surface map of the Green River Basin lower Tertiary aquifer system, 2010–14, in the northern Green River structural basin

    Science.gov (United States)

    Bartos, Timothy T.; Hallberg, Laura L.; Eddy-Miller, Cheryl

    2015-07-14

    In cooperation with the Bureau of Land Management, groundwater levels in wells located in the northern Green River Basin in Wyoming, an area of ongoing energy development, were measured by the U.S. Geological Survey from 2010 to 2014. The wells were completed in the uppermost aquifers of the Green River Basin lower Tertiary aquifer system, which is a complex regional aquifer system that provides water to most wells in the area. Except for near perennial streams, groundwater-level altitudes in most aquifers generally decreased with increasing depth, indicating a general downward potential for groundwater movement in the study area. Drilled depth of the wells was observed as a useful indicator of depth to groundwater such that deeper wells typically had a greater depth to groundwater. Comparison of a subset of wells included in this study that had historical groundwater levels that were measured during the 1960s and 1970s and again between 2012 and 2014 indicated that, overall, most of the wells showed a net decline in groundwater levels.

  19. Effects of the nanotopographic surface structure of commercially pure titanium following anodization-hydrothermal treatment on gene expression and adhesion in gingival epithelial cells.

    Science.gov (United States)

    Takebe, J; Miyata, K; Miura, S; Ito, S

    2014-09-01

    The long-term stability and maintenance of endosseous implants with anodized-hydrothermally treated commercially pure titanium surfaces and a nanotopographic structure (SA-treated c.p.Ti) depend on the barrier function provided by the interface between the transmucosal portion of the implant surface and the peri-implant epithelium. This study investigated the effects of extracellular and intracellular gene expression in adherent gingival epithelial cells cultured for 1-7 days on SA-treated c.p.Ti implant surfaces compared to anodic oxide (AO) c.p.Ti and c.p.Ti disks. Scanning electron microscopy (SEM) showed filopodium-like extensions bound closely to the nanotopographic structure of SA-treated c.p.Ti at day 7 of culture. Gene expressions of focal adhesion kinase, integrin-α6β4, and laminin-5 (α3, β3, γ2) were significantly higher on SA-treated c.p.Ti than on c.p.Ti or AO c.p.Ti after 7 days (Pcells adhere to SA-treated c.p.Ti as the transmucosal portion of an implant, and that this interaction markedly improves expression of focal adhesion molecules and enhances the epithelial cell phenotype. The cellular gene expression responses driving extracellular and intracellular molecular interactions thus play an important role in maintenance at the interface between SA-treated c.p.Ti implant surfaces and the gingival epithelial cells. Copyright © 2014 Elsevier B.V. All rights reserved.

  20. The importance of 1,2-dithiolane structure in α-lipoic acid for the downregulation of cell surface β1-integrin expression of human bladder cancer cells.

    Science.gov (United States)

    Yamasaki, Masao; Soda, Shozen; Sakakibara, Yoichi; Suiko, Masahito; Nishiyama, Kazuo

    2014-01-01

    Here, we show that cell surface β1-integrin expression, cell adhesion to fibronectin, migration, and invasion were all significantly inhibited by α-lipoic acid. These effects were not observed when cells were treated with dihydrolipoic acid or caprylic acid. These data reveal that the 1,2-dithiolane structure plays an important role in the action of α-lipoic acid.

  1. Hepatitis C Virus Activates a Neuregulin-Driven Circuit to Modify Surface Expression of Growth Factor Receptors of the ErbB Family.

    Directory of Open Access Journals (Sweden)

    Sabine Stindt

    Full Text Available Recently, the epidermal growth factor (EGF receptor (EGFR, a member of the ErbB receptor family, and its down-stream signalling have been identified as co-factors for HCV entry and replication. Since EGFR also functions as a heterodimer with other ErbB receptor family members, the subject of the present study was to investigate a possible viral interference with these cellular components. By using genotype 1b replicon cells as well as an infection-based system we found that while transcript and protein levels of EGFR and ErbB2 were up-regulated or unaffected, respectively, HCV induced a substantial reduction of ErbB3 and ErbB4 expression. Down-regulation of ErbB3 expression by HCV involves specificity protein (Sp1-mediated induction of Neuregulin (NRG1 expression as well as activation of Akt. Consistently, at transcript level disruption of ErbB3 expression by HCV can be prevented by knockdown of NRG1 or Sp1 expression, whereas reconstitution of ErbB3 protein levels requires inhibition of HCV-induced NRG1 expression and of Akt activity. Interestingly, the NRG1-mediated suppression of ErbB3 expression by HCV results in an enhanced expression of EGFR and ErbB2 on the cell surface, which can be mimicked by siRNA-mediated knockdown of ErbB3 expression. These data delineate a novel mechanism enabling HCV to sway the composition of the ErbB family members on the surface of its host cell by an NRG1-driven circuit and unravels a yet unknown cross-regulation between ErbB3 and the two other family members ErbB2 and EGFR. The shift of the receptor surface expression of the ErbB family towards enhanced expression of ErbB2 and EGFR triggered by HCV was found to promote viral RNA replication and infectivity. This suggests that HCV rearranges expression of ErbB family members to adapt the cellular environment to its requirements.

  2. Genomic Methylation Inhibits Expression of Hepatitis B Virus Envelope Protein in Transgenic Mice: A Non-Infectious Mouse Model to Study Silencing of HBV Surface Antigen Genes.

    Science.gov (United States)

    Graumann, Franziska; Churin, Yuri; Tschuschner, Annette; Reifenberg, Kurt; Glebe, Dieter; Roderfeld, Martin; Roeb, Elke

    2015-01-01

    The Hepatitis B virus genome persists in the nucleus of virus infected hepatocytes where it serves as template for viral mRNA synthesis. Epigenetic modifications, including methylation of the CpG islands contribute to the regulation of viral gene expression. The present study investigates the effects of spontaneous age dependent loss of hepatitis B surface protein- (HBs) expression due to HBV-genome specific methylation as well as its proximate positive effects in HBs transgenic mice. Liver and serum of HBs transgenic mice aged 5-33 weeks were analyzed by Western blot, immunohistochemistry, serum analysis, PCR, and qRT-PCR. From the third month of age hepatic loss of HBs was observed in 20% of transgenic mice. The size of HBs-free area and the relative number of animals with these effects increased with age and struck about 55% of animals aged 33 weeks. Loss of HBs-expression was strongly correlated with amelioration of serum parameters ALT and AST. In addition lower HBs-expression went on with decreased ER-stress. The loss of surface protein expression started on transcriptional level and appeared to be regulated epigenetically by DNA methylation. The amount of the HBs-expression correlated negatively with methylation of HBV DNA in the mouse genome. Our data suggest that methylation of specific CpG sites controls gene expression even in HBs-transgenic mice with truncated HBV genome. More important, the loss of HBs expression and intracellular aggregation ameliorated cell stress and liver integrity. Thus, targeted modulation of HBs expression may offer new therapeutic approaches. Furthermore, HBs-transgenic mice depict a non-infectious mouse model to study one possible mechanism of HBs gene silencing by hypermethylation.

  3. Expression of surface markers on the human monocytic leukaemia cell line, THP-1, as indicators for the sensitizing potential of chemicals.

    Science.gov (United States)

    An, Susun; Kim, Seoyoung; Huh, Yong; Lee, Tae Ryong; Kim, Han-Kon; Park, Kui-Lea; Eun, Hee Chul

    2009-04-01

    Evaluation of skin sensitization potential is an important part of the safety assessment of cosmetic ingredients and topical drugs. Recently, evaluation of changes in surface marker expression induced in dendritic cells (DC) or DC surrogate cell lines following exposure to chemicals represents one approach for in vitro test methods. The study aimed to test the change of expression patterns of surface markers on THP-1 cells by chemicals as a predictive in vitro method for contact sensitization. We investigated the expression of CD54, CD86, CD83, CD80, and CD40 after a 1-day exposure to sensitizers (1-chloro-2,4-dinitrobenzene; 2,4-dinitrofluorobenzene; benzocaine; 5-chloro-2-methyl-4-isothiazolin-3-one; hexyl cinnamic aldehyde; eugenol; nickel sulfate hexahydrate; potassium dichromate; cobalt sulfate; 2-mercaptobenzothiazole; and ammonium tetrachloroplatinate) and non-sensitizers (sodium lauryl sulfate, benzalkonium chloride, lactic acid, salicylic acid, isopropanol, and dimethyl sulphoxide). The test concentrations were 0.1x, 0.5x, and 1x of the 50% inhibitory concentration, and the relative fluorescence intensity was used as an expression indicator. By evaluating the expression patterns of CD54, CD86, and CD40, we could classify the chemicals as sensitizers or non-sensitizers, but CD80 and CD83 showed non-specific patterns of expression. These data suggest that the THP-1 cells are good model for screening contact sensitizers and CD40 could be a useful marker complementary to CD54 and CD86.

  4. Expression of cell adhesion and differentiation related genes in MC3T3 osteoblasts plated on titanium alloys: role of surface properties

    International Nuclear Information System (INIS)

    Sista, Subhash; Wen, Cuie; Hodgson, Peter D.; Pande, Gopal

    2013-01-01

    It is important to understand the cellular and molecular events that take place at the cell–material interface of implants used for bone repair. An understanding of the mechanisms involved in the initial stages of osteoblast interactions with the surface of the implant material is fundamental in deciding the fate of the cells that come in contact with it. In this study, we compared the relative gene expression of markers that are known to be associated with cell adhesion and differentiation in MC3T3 osteoblast cells, at various time points after plating the cells on surfaces of titanium (Ti) and its two alloys, titanium–zirconium (TiZr) and titanium–niobium (TiNb) by using Quantitative Real Time Polymerase Chain Reaction (RT-PCR). Our analysis indicated that expression of adhesion supporting genes was higher on TiZr surface as compared to Ti and TiNb. The behavior of these genes is possibly driven by a higher surface energy of TiZr. However no significant difference in the expression of differentiation related genes could be seen between the two alloys, although on both substrates it was higher as compared to unalloyed Ti. We propose that substrate composition of the alloys can influence the adhesion and differentiation related gene expression and that Ti alloys are better substrates for inducing osteogenesis as compared to unalloyed Ti. - Highlights: ► Methodology for comparing gene expression in osteoblasts plated on Ti, TiZr or TiNb ► Alloys with higher surface energy (TiZr) induce cell adhesion genes more efficiently ► Alloyed Ti is superior to unalloyed Ti to induce osteoblast differentiation genes

  5. Expression of CD64 on Surface of Circulating Monocytes in Systemic Lupus Erythematosus Patients: Relation to Disease Activity and Lupus Nephritis.

    Science.gov (United States)

    Abd-Elhamid, Yasser A; Eltanawy, Refaat M; Fawzy, Rasha M; Fouad, Nehad A; Atlm, Ann M

    2017-01-01

    CD64 is a type of integral membrane glycoprotein known as FC receptor that binds monomeric IgG-type antibodies with high affinity. It is more commonly known as FC gamma receptor 1 (FC?R1) and it is expressed on monocytes surface. The goal of this study was to investigate the association of CD64 expression on the surface of peripheral blood monocytes of systemic lupus erythematosus patients with disease activity, and lupus nephritis. 30 SLE patients were enrolled into this study. They were subdivided into: 15 SLE patients with lupus nephritis and 15 SLE patients without lupus nephritis. In addition,Together with 25 age and sex matched healthy volunteers were enrolled as controls group. Disease activity was defined by SLE Disease Activity Index (SLEDAI) score and the renal Systemic Lupus Erythematosus Disease Activity Index (rSLEDAI) score. Surface expression of CD64 on peripheral blood monocytes was evaluated by Flowcytometry. Renal biopsies of Lupus nephritis patients was obtained and evaluated using the International Society of Nephrology/Renal Pathology Society (ISN/RPS) classification scheme. There was a statistically significant difference in surface expression of CD64 on circulating monocytes (P > 0.001) in SLE patients with nephritis especially those with class II/III as compared to SLE without nephritis and healthy controls. The mean fluorescent intensity of CD64 staining correlated positively with markers of systemic inflammation, lupus nephritis, SLEDAI and rSLEDAI scores. In conclusions, surface expression of CD64 on circulating monocytes reflects systemic inflammation, renal injury and could be used as a rapid approach and good biomarker for disease activity and lupus nephritis in SLE patients. Copyright© by the Egyptian Association of Immunologists.

  6. Effects of calcium ion incorporation on osteoblast gene expression in MC3T3-E1 cells cultured on microstructured titanium surfaces.

    Science.gov (United States)

    Park, Jin-Woo; Suh, Jo-Young; Chung, Hyun-Ju

    2008-07-01

    The surface characteristics of a calcium ion (Ca)-incorporated titanium (Ti) surface, produced by hydrothermal treatment using an alkaline Ca-containing solution, and its effects on osteoblastic differentiation were investigated. MC3T3-E1 pre-osteoblastic cells were cultured on machined or grit-blasted Ti surfaces with and without Ca incorporation. The MTT assay was used to determine cell proliferation, and real-time PCR was used for quantitative analysis of osteoblastic gene expression. Hydrothermal treatment with a Ca-containing solution produced a crystalline CaTiO(3) nanostructure of approximately 100 nm in dimension, preserving original micron-scaled surface topographies and microroughness caused by machining, blasting, or blasting and etching treatments. After immersion in Hank's balanced salt solution, considerable apatite formation was observed on all surfaces of the Ca-incorporated samples. Significantly more cell proliferation was found on Ca-incorporated Ti surfaces than on untreated Ti surfaces (p < 0.001). Quantitative real-time PCR analysis showed notably higher alkaline phosphatase, osteopontin, and osteocalcin mRNA levels in cells grown on Ca-incorporated blasted surfaces than on other surfaces at an early time point. Thus, Ca incorporation may have a beneficial effect on osseointegration of microstructured Ti implants by accelerating osteoblast proliferation and differentiation during the early healing phase following implantation. (c) 2007 Wiley Periodicals, Inc.

  7. Comprehensive evaluation of dioxins and dioxin-like compounds in surface soils and river sediments from e-waste-processing sites in a village in northern Vietnam: Heading towards the environmentally sound management of e-waste

    OpenAIRE

    Go Suzuki; Masayuki Someya; Hidenori Matsukami; Nguyen Minh Tue; Natsuyo Uchida; Le Huu Tuyen; Pham Hung Viet; Shin Takahashi; Shinsuke Tanabe; Abraham Brouwer; Hidetaka Takigami

    2016-01-01

    The management of electronic waste (e-waste), which can be a source of both useful materials and toxic substances, depending on the processing method, is important for promoting material cycling. In this study, we used the dioxin-responsive chemical-activated luciferase gene expression (DR-CALUX) assay combined with gas chromatography–high-resolution mass spectrometry to evaluate the levels of dioxin-like compounds in surface soils and river sediments collected in and around an e-waste-proces...

  8. Evaluating bacterial community structures in oil collected from the sea surface and sediment in the northern Gulf of Mexico after the Deepwater Horizon oil spill

    OpenAIRE

    Liu, Zhanfei; Liu, Jiqing

    2013-01-01

    Bacterial community structures were evaluated in oil samples using culture-independent pyrosequencing, including oil mousses collected on sea surface and salt marshes during the Deepwater Horizon oil spill, and oil deposited in sediments adjacent to the wellhead 1 year after the spill. Phylogenetic analysis suggested that Erythrobacter, Rhodovulum, Stappia, and Thalassospira of Alphaproteobacteria were the prevailing groups in the oil mousses, which may relate to high temperatures and strong ...

  9. Transmembrane and ubiquitin-like domain-containing protein 1 (Tmub1/HOPS facilitates surface expression of GluR2-containing AMPA receptors.

    Directory of Open Access Journals (Sweden)

    Hyunjeong Yang

    Full Text Available Some ubiquitin-like (UBL domain-containing proteins are known to play roles in receptor trafficking. Alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptors (AMPARs undergo constitutive cycling between the intracellular compartment and the cell surface in the central nervous system. However, the function of UBL domain-containing proteins in the recycling of the AMPARs to the synaptic surface has not yet been reported.Here, we report that the Transmembrane and ubiquitin-like domain-containing 1 (Tmub1 protein, formerly known as the Hepatocyte Odd Protein Shuttling (HOPS protein, which is abundantly expressed in the brain and which exists in a synaptosomal membrane fraction, facilitates the recycling of the AMPAR subunit GluR2 to the cell surface. Neurons transfected with Tmub1/HOPS-RNAi plasmids showed a significant reduction in the AMPAR current as compared to their control neurons. Consistently, the synaptic surface expression of GluR2, but not of GluR1, was significantly decreased in the neurons transfected with the Tmub1/HOPS-RNAi and increased in the neurons overexpressing EGFP-Tmub1/HOPS. The altered surface expression of GluR2 was speculated to be due to the altered surface-recycling of the internalized GluR2 in our recycling assay. Eventually, we found that GluR2 and glutamate receptor interacting protein (GRIP were coimmunoprecipitated by the anti-Tmub1/HOPS antibody from the mouse brain. Taken together, these observations show that the Tmub1/HOPS plays a role in regulating basal synaptic transmission; it contributes to maintain the synaptic surface number of the GluR2-containing AMPARs by facilitating the recycling of GluR2 to the plasma membrane.

  10. Substance P Increases Cell-Surface Expression of CD74 (Receptor for Macrophage Migration Inhibitory Factor: In Vivo Biotinylation of Urothelial Cell-Surface Proteins

    Directory of Open Access Journals (Sweden)

    Katherine L. Meyer-Siegler

    2009-01-01

    N-hydroxysulfosuccinimide biotin ester-labeled surface urothelial proteins in rats treated either with saline or substance P (SP, 40 μg/kg. The bladder was examined by histology and confocal microscopy. Biotinylated proteins were purified by avidin agarose, immunoprecipitated with anti-MIF or anti-CD74 antibodies, and detected with strepavidin-HRP. Only superficial urothelial cells were biotinylated. These cells contained a biotinylated MIF/CD74 cell-surface complex that was increased in SP-treated animals. SP treatment increased MIF and CD74 mRNA in urothelial cells. Our data indicate that intraluminal MIF, released from urothelial cells as a consequence of SP treatment, interacts with urothelial cell-surface CD74. These results document that our previously described MIF-CD74 interaction occurs at the urothelial cell surface.

  11. Study of the impact of radionuclides discharges from nuclear industry in the northern Cotentin area on the oyster Crassostrea gigas: expression of cellular stress molecular markers

    International Nuclear Information System (INIS)

    Farcy, E.

    2006-11-01

    This thesis explores the potential radiological impact of radionuclide discharges from the nuclear industry on the Pacific oyster, Crassostrea gigas. One of the major goals of this research was to identify markers that could be used to monitor the effects of low-level chronic irradiation. We decided to focalize on the expression of stress-induced genes involved in the regulation of cellular stress, focusing on transcription. First, homology cloning was used to identify four new cDNAs encoding stress markers. Then data collected at various sites enabled to evidence that mRNA levels for each of the genes of interest naturally vary to a significant degree, based on individual differences and seasonal factors. Comparing oysters from exposed sites with those from a reference site located on the Atlantic coast did not suggest any relationship between mRNA levels changes and the oysters exposure to liquid radioactive waste from the A.R.E.V.A. reprocessing plant. In the environment, we found that those radionuclide releases resulted in a very small increase in radioactivity in oysters, especially compared with their natural radioactivity. In the laboratory, by exposing the oysters to higher radionuclide concentrations than those found in the environment, we were able to identify two genes as potential candidates for studying the effects of chronic exposure to low doses of ionizing radiations in the oyster: genes encoding MT and MXR. We confirmed that transcriptional induction of these two genes occurs in response to high doses of acute irradiation. Finally, the comparison between environmental results and the laboratory results underlined the complementarity of these two approaches. In particular, it revealed that seasonal variations in mRNA levels probably have a significant effect on the stress response. (author)

  12. Linkage between seasonal insolation gradient in the tropical northern hemisphere and the sea surface salinity of the equatorial Indian Ocean during the last glacial period

    Digital Repository Service at National Institute of Oceanography (India)

    Saraswat, R.; Nigam, R.; Mackensen, A.; Weldeab, S.

    oxygen isotopic composition to estimate the local seawater oxygen isotopic composition (δ18Osw-ivc). The change in δ18Osw-ivc relative to the present day value (∆δ18Osw-ivc) was then calculated by subtracting the modern surface seawater oxygen isotopic... the modern measurement (Fig. 5), match very well with the Levitus 7 annual SSS over the core site, showing values of 34.7 and 34.8, respectively. Delaygue et al (2001) also developed a seawater oxygen isotope/salinity relationship for the Bay of Bengal...

  13. Surface co-expression of two different PfEMP1 antigens on single Plasmodium falciparum-infected erythrocytes facilitates binding to ICAM1 and PECAM1

    DEFF Research Database (Denmark)

    Joergensen, Louise; Bengtsson, Dominique C; Bengtsson, Anja

    2010-01-01

    gene is expressed per cell at a time. We measured var mRNA transcript levels by real-time Q-PCR, analysed var gene transcripts by single-cell FISH and directly compared these with PfEMP1 antigen surface expression and cytoadhesion in three different antibody-selected P. falciparum 3D7 sub-lines using...... was found to bind both CD31/PECAM1 and CD54/ICAM1 and to adhere twice as efficiently to human endothelial cells, compared to infected cells having only one PfEMP1 variant on the surface. These new results on PfEMP1 antigen expression indicate that a re-evaluation of the molecular mechanisms involved in P...... live confocal microscopy, flow cytometry and in vitro adhesion assays. We found that one selected parasite sub-line simultaneously expressed two different var genes as surface antigens, on single IE. Importantly, and of physiological relevance to adhesion and malaria pathogenesis, this parasite sub-line...

  14. Inhibitory effects of two G protein-coupled receptor kinases on the cell surface expression and signaling of the human adrenomedullin receptor

    International Nuclear Information System (INIS)

    Kuwasako, Kenji; Sekiguchi, Toshio; Nagata, Sayaka; Jiang, Danfeng; Hayashi, Hidetaka; Murakami, Manabu; Hattori, Yuichi; Kitamura, Kazuo; Kato, Johji

    2016-01-01

    Receptor activity-modifying protein 2 (RAMP2) enables the calcitonin receptor-like receptor (CLR, a family B GPCR) to form the type 1 adrenomedullin receptor (AM 1 receptor). Here, we investigated the effects of the five non-visual GPCR kinases (GRKs 2 through 6) on the cell surface expression of the human (h)AM 1 receptor by cotransfecting each of these GRKs into HEK-293 cells that stably expressed hRAMP2. Flow cytometric analysis revealed that when coexpressed with GRK4 or GRK5, the cell surface expression of the AM 1 receptor was markedly decreased prior to stimulation with AM, thereby attenuating both the specific [ 125 I]AM binding and AM-induced cAMP production. These inhibitory effects of both GRKs were abolished by the replacement of the cytoplasmic C-terminal tail (C-tail) of CLR with that of the calcitonin receptor (a family B GPCR) or β 2 -adrenergic receptor (a family A GPCR). Among the sequentially truncated CLR C-tail mutants, those lacking the five residues 449–453 (Ser-Phe-Ser-Asn-Ser) abolished the inhibition of the cell surface expression of CLR via the overexpression of GRK4 or GRK5. Thus, we provided new insight into the function of GRKs in agonist-unstimulated GPCR trafficking using a recombinant AM 1 receptor and further determined the region of the CLR C-tail responsible for this GRK function. - Highlights: • We discovered a novel function of GRKs in GPCR trafficking using human CLR/RAMP2. • GRKs 4 and 5 markedly inhibited the cell surface expression of human CLR/RAMP2. • Both GRKs exhibited highly significant receptor signaling inhibition. • Five residues of the C-terminal tail of CLR govern this function of GRKs.

  15. Inhibitory effects of two G protein-coupled receptor kinases on the cell surface expression and signaling of the human adrenomedullin receptor

    Energy Technology Data Exchange (ETDEWEB)

    Kuwasako, Kenji, E-mail: kuwasako@med.miyazaki-u.ac.jp [Frontier Science Research Center, University of Miyazaki, Miyazaki, 889-1692 (Japan); Sekiguchi, Toshio [Noto Marine Laboratory, Division of Marine Environmental Studies, Institute of Nature and Environmental Technology, Kanazawa University, Ishikawa, 927-0553 (Japan); Nagata, Sayaka [Division of Circulatory and Body Fluid Regulation, Faculty of Medicine, University of Miyazaki, Miyazaki, 889-1692 (Japan); Jiang, Danfeng; Hayashi, Hidetaka [Frontier Science Research Center, University of Miyazaki, Miyazaki, 889-1692 (Japan); Murakami, Manabu [Department of Pharmacology, Hirosaki University, Graduate School of Medicine, Hirosaki, 036-8562 (Japan); Hattori, Yuichi [Department of Molecular and Medical Pharmacology, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, Toyama, 930-0194 (Japan); Kitamura, Kazuo [Division of Circulatory and Body Fluid Regulation, Faculty of Medicine, University of Miyazaki, Miyazaki, 889-1692 (Japan); Kato, Johji [Frontier Science Research Center, University of Miyazaki, Miyazaki, 889-1692 (Japan)

    2016-02-19

    Receptor activity-modifying protein 2 (RAMP2) enables the calcitonin receptor-like receptor (CLR, a family B GPCR) to form the type 1 adrenomedullin receptor (AM{sub 1} receptor). Here, we investigated the effects of the five non-visual GPCR kinases (GRKs 2 through 6) on the cell surface expression of the human (h)AM{sub 1} receptor by cotransfecting each of these GRKs into HEK-293 cells that stably expressed hRAMP2. Flow cytometric analysis revealed that when coexpressed with GRK4 or GRK5, the cell surface expression of the AM{sub 1} receptor was markedly decreased prior to stimulation with AM, thereby attenuating both the specific [{sup 125}I]AM binding and AM-induced cAMP production. These inhibitory effects of both GRKs were abolished by the replacement of the cytoplasmic C-terminal tail (C-tail) of CLR with that of the calcitonin receptor (a family B GPCR) or β{sub 2}-adrenergic receptor (a family A GPCR). Among the sequentially truncated CLR C-tail mutants, those lacking the five residues 449–453 (Ser-Phe-Ser-Asn-Ser) abolished the inhibition of the cell surface expression of CLR via the overexpression of GRK4 or GRK5. Thus, we provided new insight into the function of GRKs in agonist-unstimulated GPCR trafficking using a recombinant AM{sub 1} receptor and further determined the region of the CLR C-tail responsible for this GRK function. - Highlights: • We discovered a novel function of GRKs in GPCR trafficking using human CLR/RAMP2. • GRKs 4 and 5 markedly inhibited the cell surface expression of human CLR/RAMP2. • Both GRKs exhibited highly significant receptor signaling inhibition. • Five residues of the C-terminal tail of CLR govern this function of GRKs.

  16. Effects of the nanotopographic surface structure of commercially pure titanium following anodization–hydrothermal treatment on gene expression and adhesion in gingival epithelial cells

    International Nuclear Information System (INIS)

    Takebe, J.; Miyata, K.; Miura, S.; Ito, S.

    2014-01-01

    The long-term stability and maintenance of endosseous implants with anodized–hydrothermally treated commercially pure titanium surfaces and a nanotopographic structure (SA-treated c.p.Ti) depend on the barrier function provided by the interface between the transmucosal portion of the implant surface and the peri-implant epithelium. This study investigated the effects of extracellular and intracellular gene expression in adherent gingival epithelial cells cultured for 1–7 days on SA-treated c.p.Ti implant surfaces compared to anodic oxide (AO) c.p.Ti and c.p.Ti disks. Scanning electron microscopy (SEM) showed filopodium-like extensions bound closely to the nanotopographic structure of SA-treated c.p.Ti at day 7 of culture. Gene expressions of focal adhesion kinase, integrin-α6β4, and laminin-5 (α3, β3, γ2) were significantly higher on SA-treated c.p.Ti than on c.p.Ti or AO c.p.Ti after 7 days (P < 0.05). Our results confirmed that gingival epithelial cells adhere to SA-treated c.p.Ti as the transmucosal portion of an implant, and that this interaction markedly improves expression of focal adhesion molecules and enhances the epithelial cell phenotype. The cellular gene expression responses driving extracellular and intracellular molecular interactions thus play an important role in maintenance at the interface between SA-treated c.p.Ti implant surfaces and the gingival epithelial cells. - Highlights: • SA-treated Ti provides a nanotopographic structure for clinical oral implants. • This could regulate integrin-mediated epithelial cell adhesion and gene expression. • FAK mRNA was significantly higher on SA-treated Ti. • Integrin-α6β4 and laminin-5 mRNA were significantly higher on SA-treated Ti. • Extracellular/intracellular molecular interactions play a key role on SA-treated Ti

  17. A methodology for analysing temporal changes of forest surface using aerial photos for the application of Kyoto protocol: a study case in Northern Italy

    Directory of Open Access Journals (Sweden)

    Salvadori I

    2006-01-01

    Full Text Available According to art. 3.3 of the Kyoto Protocol, Italy, like other Parties included in Annex I, shall report the net changes in greenhouse gas emissions by sources and removals by sinks resulting from afforestation, reforestation and deforestation activities (ARD. To evaluate these activities, Italy has to elaborate methods to estimate the conversion of non-forested to forested land, occurred after 31 December 1989. The aim of this study was to test a methodology to estimate the ARD activities. The approach was experimented in the Comunità Montana del Grappa (about 10500 ha considered as a pilot area in the Prealpine region (NE Italy. The land-use change relative to the forest area was assessed by multitemporal classification of 1131 sampling points on orthocorrected aerial photos relative to 1991, 1996 and 1999. The forest area based on different definitions (minimum land cover equal to 10%, 20% or 30% and minimum surface equal to 2000 m2 or 5000 m2 was also assessed. Between 1991 and 1999, the total increment of the forest area was equal to 224 ha. However, the estimated increment was strongly related to the minimum surface (2000 m2 vs 5000 m2 of the forest definition. The proposed procedure was relatively easy to implement and highlighted the role of ARD and revegetation to attain the goals appointed from the Kyoto Protocol.

  18. Evaluating bacterial community structures in oil collected from the sea surface and sediment in the northern Gulf of Mexico after the Deepwater Horizon oil spill.

    Science.gov (United States)

    Liu, Zhanfei; Liu, Jiqing

    2013-06-01

    Bacterial community structures were evaluated in oil samples using culture-independent pyrosequencing, including oil mousses collected on sea surface and salt marshes during the Deepwater Horizon oil spill, and oil deposited in sediments adjacent to the wellhead 1 year after the spill. Phylogenetic analysis suggested that Erythrobacter, Rhodovulum, Stappia, and Thalassospira of Alphaproteobacteria were the prevailing groups in the oil mousses, which may relate to high temperatures and strong irradiance in surface Gulf waters. In the mousse collected from the leaves of Spartina alterniflora, Vibrio of Gammaproteobacteria represented 57% of the total operational taxonomic units, suggesting that this indigenous genus is particularly responsive to the oil contamination in salt marshes. The bacterial communities in oil-contaminated sediments were highly diversified. The relatively high abundance of the Methylococcus, Methylobacter, Actinobacteria, Firmicutes, and Chlorofexi bacteria resembles those found in certain cold-seep sediments with gas hydrates. Bacterial communities in the overlying water of the oil-contaminated sediment were dominated by Ralstonia of Betaproteobacteria, which can degrade small aromatics, and Saccharophagus degradans of Gammaproteobacteria, a cellulose degrader, suggesting that overlying water was affected by the oil-contaminated sediments, possibly due to the dissolution of small aromatics and biosurfactants produced during biodegradation. Overall, these results provided key information needed to evaluate oil degradation in the region and develop future bioremediation strategies. © 2013 The Authors. Microbiology Open published by John Wiley & Sons Ltd.

  19. Establishment of insect cell lines expressing green fluorescent protein on cell surface based on AcMNPV GP64 membrane fusion characteristic.

    Science.gov (United States)

    Qi, Ben-Xiang; Chen, Ying-Jian; Su, Rui; Li, Yi-Fei; Zheng, Gui-Ling; Li, Chang-You

    2017-10-01

    Displaying a protein on the surface of cells has been provided a very successful strategy to function research of exogenous proteins. Based on the membrane fusion characteristic of Autographa californica multiple nucleopolyhedrovirus envelope protein GP64, we amplified and cloned N-terminal signal peptide and C-terminal transmembrane domain as well as cytoplasmic tail domain of gp64 gene into vector pIZ/V5-His with multi-cloning sites to construct the cell surface expression vector pIZ/V5-gp64. To verify that the vector can be used to express proteins on the membrane of insect cells, a recombinant plasmid pIZ/V5-gp64-GFP was constructed by introducing the PCR amplified green fluorescent protein (GFP) gene and transfected into insect cell lines Sf9 and H5. The transected cells were screened with zeocin and cell cloning. PCR verification results showed that the GFP gene was successfully integrated into these cells. Green fluorescence in Sf9-GFP and H5-GFP cells was observed by using confocal laser scanning microscopy and immunofluorescence detection indicated that GFP protein was located on the cell membrane. Western blot results showed that a fusion protein GP64-GFP of about 40 kDa was expressed on the membrane of Sf9-GFP and H5-GFP cells. The expression system constructed in this paper can be used for localization and continuous expression of exogenous proteins on insect cell membrane.

  20. The Plasmodium falciparum transcriptome in severe malaria reveals altered expression of genes involved in important processes including surface antigen–encoding var genes

    Science.gov (United States)

    Tonkin-Hill, Gerry Q.; Trianty, Leily; Noviyanti, Rintis; Nguyen, Hanh H. T.; Sebayang, Boni F.; Lampah, Daniel A.; Marfurt, Jutta; Cobbold, Simon A.; Rambhatla, Janavi S.; McConville, Malcolm J.; Rogerson, Stephen J.; Brown, Graham V.; Day, Karen P.; Price, Ric N.; Anstey, Nicholas M.

    2018-01-01

    Within the human host, the malaria parasite Plasmodium falciparum is exposed to multiple selection pressures. The host environment changes dramatically in severe malaria, but the extent to which the parasite responds to—or is selected by—this environment remains unclear. From previous studies, the parasites that cause severe malaria appear to increase expression of a restricted but poorly defined subset of the PfEMP1 variant, surface antigens. PfEMP1s are major targets of protective immunity. Here, we used RNA sequencing (RNAseq) to analyse gene expression in 44 parasite isolates that caused severe and uncomplicated malaria in Papuan patients. The transcriptomes of 19 parasite isolates associated with severe malaria indicated that these parasites had decreased glycolysis without activation of compensatory pathways; altered chromatin structure and probably transcriptional regulation through decreased histone methylation; reduced surface expression of PfEMP1; and down-regulated expression of multiple chaperone proteins. Our RNAseq also identified novel associations between disease severity and PfEMP1 transcripts, domains, and smaller sequence segments and also confirmed all previously reported associations between expressed PfEMP1 sequences and severe disease. These findings will inform efforts to identify vaccine targets for severe malaria and also indicate how parasites adapt to—or are selected by—the host environment in severe malaria. PMID:29529020

  1. Decreased surface expression of the δ subunit of the GABAA receptor contributes to reduced tonic inhibition in dentate granule cells in a mouse model of fragile X syndrome.

    Science.gov (United States)

    Zhang, Nianhui; Peng, Zechun; Tong, Xiaoping; Lindemeyer, A Kerstin; Cetina, Yliana; Huang, Christine S; Olsen, Richard W; Otis, Thomas S; Houser, Carolyn R

    2017-11-01

    While numerous changes in the GABA system have been identified in models of Fragile X Syndrome (FXS), alterations in subunits of the GABA A receptors (GABA A Rs) that mediate tonic inhibition are particularly intriguing. Considering the key role of tonic inhibition in controlling neuronal excitability, reduced tonic inhibition could contribute to FXS-associated disorders such as hyperactivity, hypersensitivity, and increased seizure susceptibility. The current study has focused on the expression and function of the δ subunit of the GABA A R, a major subunit involved in tonic inhibition, in granule cells of the dentate gyrus in the Fmr1 knockout (KO) mouse model of FXS. Electrophysiological studies of dentate granule cells revealed a marked, nearly four-fold, decrease in tonic inhibition in the Fmr1 KO mice, as well as reduced effects of two δ subunit-preferring pharmacological agents, THIP and DS2, supporting the suggestion that δ subunit-containing GABA A Rs are compromised in the Fmr1 KO mice. Immunohistochemistry demonstrated a small but statistically significant decrease in δ subunit labeling in the molecular layer of the dentate gyrus in Fmr1 KO mice compared to wildtype (WT) littermates. The discrepancy between the large deficits in GABA-mediated tonic inhibition in granule cells in the Fmr1 KO mice and only modest reductions in immunolabeling of the δ subunit led to studies of surface expression of the δ subunit. Cross-linking experiments followed by Western blot analysis demonstrated a small, non-significant decrease in total δ subunit protein in the hippocampus of Fmr1 KO mice, but a four-fold decrease in surface expression of the δ subunit in these mice. No significant changes were observed in total or surface expression of the α4 subunit protein, a major partner of the δ subunit in the forebrain. Postembedding immunogold labeling for the δ subunit demonstrated a large, three-fold, decrease in the number of symmetric synapses with

  2. Transforming Growth Factor-β2 Downregulates Major Histocompatibility Complex (MHC) I and MHC II Surface Expression on Equine Bone Marrow-Derived Mesenchymal Stem Cells Without Altering Other Phenotypic Cell Surface Markers.

    Science.gov (United States)

    Berglund, Alix K; Fisher, Matthew B; Cameron, Kristin A; Poole, Emma J; Schnabel, Lauren V

    2017-01-01

    Allogeneic mesenchymal stem cells (MSCs) are a promising cell source for treating musculoskeletal injuries in horses. Effective and safe allogeneic therapy may be hindered, however, by recipient immune recognition and rejection of major histocompatibility complex (MHC)-mismatched MSCs. Development of strategies to prevent immune rejection of MHC-mismatched MSCs in vivo is necessary to enhance cell survival and potentially increase the efficacy and safety of allogeneic MSC therapy. The purposes of this study were to evaluate if transforming growth factor-β2 (TGF-β2) downregulated MHC expression on equine MSCs and to determine if TGF-β2 treatment altered the phenotype of MSCs. Equine bone marrow-derived MSCs from 12 horses were treated with 1, 5, or 10 ng/ml TGF-β2 from initial isolation until MHC expression analysis. TGF-β2-treated MSCs had reduced MHC I and MHC II surface expression compared to untreated controls. TGF-β2 treatment also partially blocked IFN-γ-induced upregulation of MHC I and MHC II. Constitutive and IFN-γ-induced MHC I and MHC II expression on equine MSCs was dynamic and highly variable, and the effect of TGF-β2 was significantly dependent on the donor animal and baseline MHC expression. TGF-β2 treatment did not appear to change morphology, surface marker expression, MSC viability, or secretion of TGF-β1, but did significantly increase the number of cells obtained from culture. These results indicate that TGF-β2 treatment has promise for regulating MHC expression on MSCs to facilitate allogeneic therapy, but further work is needed to maintain MHC stability when exposed to an inflammatory stimulus.

  3. Allelic imbalance modulates surface expression of the tolerance-inducing HLA-G molecule on primary trophoblast cells

    DEFF Research Database (Denmark)

    Djurisic, S; Teiblum, S; Tolstrup, C K

    2015-01-01

    The HLA-G molecule is expressed on trophoblast cells at the feto-maternal interface, where it interacts with local immune cells, and upholds tolerance against the semi-allogeneic fetus. Aberrant HLA-G expression in the placenta and reduced soluble HLA-G levels are observed in pregnancy complicati...

  4. Possibilities of new materials surface sensibility express determination based on ZnSe-CdS system by pH isoelectric state measurements of the surface state

    Science.gov (United States)

    Kirovskaya, I. A.; Mironova, E. V.; Ushakov, O. V.; Nor, P. E.; Yureva, A. V.; Matyash, Yu I.

    2018-01-01

    A method for determining the hydrogen index of the surfaces isoelectric state (pHiso) at various gases pressures -possible components of the surrounding and technological media has been developed. With its use, changes in pH of binary and more complex semiconductors-components of the new system-ZnSe-CdS under the influence of nitrogen dioxide-have been found. The limiting sensitivity of surfaces - minimum PNO2, causing a change in pH has been estimated. The most active components of ZnSe-CdS system, recommended as materials for measuring cells of NO2, have been revealed. The relationship between the changing patterns with the composition of surface (acid-base) and bulk (in particular, theoretical calculated crystal density) properties has been established, allowing to find the most effective materials for sensor technology and for semiconductor analysis.

  5. Genetic Variations in the Human G Protein-coupled Receptor Class C, Group 6, Member A (GPRC6A) Control Cell Surface Expression and Function

    DEFF Research Database (Denmark)

    Jorgensen, Stine; Have, Christian Theil; Underwood, Christina Rye

    2017-01-01

    GPRC6A is a G protein-coupled receptor activated by l-amino acids, which, based on analyses of knock-out mice, has been suggested to have physiological functions in metabolism and testicular function. The human ortholog is, however, mostly retained intracellularly in contrast to the cell surface...... of the human ortholog. Genetic analyses of the 1000 genome database and the Inter99 cohort of 6,000 Danes establish the distribution of genotypes among ethnic groups, showing that the cell surface-expressed and functional variant is much more prevalent in the African population than in European and Asian...

  6. Water pollution abatement programme. The Czech Republic. Project 4.2. Assessing critical loads of acidity to surface waters in The Czech Republic. Critical loads of acidity to surface waters, Northern Moravia and Silesia, The Czech Republic

    Energy Technology Data Exchange (ETDEWEB)

    Lien, L.; Raclavsky, K.; Henriksen, A.; Raclavska, H.; Matysek, D.

    1994-12-31

    The governments of Norway and Czech and Slovak Federal Republic have signed a bilateral environmental protection agreement. This report describes Project 4.2 of the agreement: Assessing critical loads of acidity to surface waters in The Czech Republic. Critical load of acidity to surface waters and exceedance of critical load were estimated by using standard methods modified for the sampling area. Water samples were mainly taken from small forest streams, which were the only available surface waters with negligible pollution from local sources. High critical loads were calculated, averaging 20 times higher than the corresponding value for southern Norway. The deposition of acidifying components in the region was high, but did not exceed the critical load and so there is a reserve for additional acid deposition. Scattered water analyses from several other parts of The Czech Republic indicate both low critical loads and exceedance of critical load in various regions (e.g. Bohemia). 21 refs., 14 figs., 3 tabs.

  7. Northern forests, Chapter 7

    Science.gov (United States)

    L.H. Pardo; C.L. Goodale; E.A. Lilleskov; L.H. Geiser

    2011-01-01

    The Northern Forests ecological region spans much of Canada, from Saskatchewan to Newfoundland; its southern portion extends into the northern United States (CEC 1997). The U.S. component includes the northern hardwood and spruce-fir forest types and encompasses parts of the Northeast (mountainous regions in Pennsylvania, New York, New Jersey, Connecticut,...

  8. Prostaglandin E2 Receptor Expression by Osteoblasts is Modulated by Implant Surface Roughness and Prostaglandin E2

    National Research Council Canada - National Science Library

    Campbell, MaCasey M

    2006-01-01

    .... Relatively little is known about the cellular receptors for prostaglandins, EP receptors, especially with regard to osteoblast response to implant surface roughness and early events preceding osseointegration...

  9. Distribution and levels of cell surface expression of CD33 and CD123 in acute myeloid leukemia

    Science.gov (United States)

    Ehninger, A; Kramer, M; Röllig, C; Thiede, C; Bornhäuser, M; von Bonin, M; Wermke, M; Feldmann, A; Bachmann, M; Ehninger, G; Oelschlägel, U

    2014-01-01

    Owing to the more recent positive results with the anti-CD33 immunotoxin gemtuzumab ozogamicin, therapy against acute myeloid leukemias (AMLs) targeting CD33 holds many promises. Here, CD33 and CD123 expression on AML blasts was studied by flow cytometry in a cohort of 319 patients with detailed information on French–American–British/World Health Organization (FAB/WHO) classification, cytogenetics and molecular aberrations. AMLs of 87.8% express CD33 and would therefore be targetable with anti-CD33 therapies. Additionally, 9.4% of AMLs express CD123 without concomitant CD33 expression. Thus, nearly all AMLs could be either targeted via CD33 or CD123. Simultaneous presence of both antigens was observed in 69.5% of patients. Most importantly, even AMLs with adverse cytogenetics express CD33 and CD123 levels comparable to those with favorable and intermediate subtypes. Some patient groups with unfavorable alterations, such as FMS-related tyrosine kinase 3-internal tandem duplication (FLT3-ITD) mutations, high FLT3-ITD mutant/wild-type ratios and monosomy 5 are even characterized by high expression of CD33 and CD123. In addition, blasts of patients with mutant nucleophosmin (NPM1) revealed significantly higher CD33 and CD123 expression pointing toward the possibility of minimal residual disease-guided interventions in mutated NPM1-positive AMLs. These results stimulate the development of novel concepts to redirect immune effector cells toward CD33- and CD123-expressing blasts using bi-specific antibodies or engineered T cells expressing chimeric antigen receptors. PMID:24927407

  10. Connecting the surface to near-shore bottom waters in the California Current ecosystem: a study of Northern California interannual to decadal oceanographic variability

    Science.gov (United States)

    Fish, C.; Hill, T. M.; Davis, C. V.; Lipski, D.; Jahncke, J.

    2017-12-01

    Elucidating both surface and bottom water ecosystem impacts of temperature change, acidification, and food web disruption are needed to understand anthropogenic processes in the ocean. The Applied California Current Ecosystem Studies (ACCESS) partnership surveys the California Current within the Greater Farallones and Cordell Bank National Marine Sanctuaries three times annually, sampling water column hydrography and discrete water samples from 0 m and 200 m depth at five stations along three primary transects. The transects span the continental shelf with stations as close as 13 km from the coastline to 65 km. This time series extends from 2004 to 2017, integrating information on climate, productivity, zooplankton abundance, oxygenation, and carbonate chemistry. We focus on the interpretation of the 2012-2017 carbonate chemistry data and present both long term trends over the duration of the time series as well as shorter term variability (e.g., ENSO, `warm blob' conditions) to investigate the region's changing oceanographic conditions. For example, we document oscillations in carbonate chemistry, oxygenation, and foraminiferal abundance in concert with interannual oceanographic variability and seasonal (upwelling) cycles. We concentrate on results from near Cordell Bank that potentially impact deep sea coral ecosystems.

  11. Bromine, iodine and sodium in surface snow along the 2013 Talos Dome-GV7 traverse (northern Victoria Land, East Antarctica)

    Science.gov (United States)

    Maffezzoli, Niccolò; Spolaor, Andrea; Barbante, Carlo; Bertò, Michele; Frezzotti, Massimo; Vallelonga, Paul

    2017-03-01

    Halogen chemistry in the polar regions occurs through the release of halogen elements from different sources. Bromine is primarily emitted from sea salt aerosols and other saline condensed phases associated with sea ice surfaces, while iodine is affected by the release of organic compounds from algae colonies living within the sea ice environment. Measurements of halogen species in polar snow samples are limited to a few sites although there is some evidence that they are related to sea ice extent. We examine here total bromine, iodine and sodium concentrations in a series of 2 m cores collected during a traverse from Talos Dome (72°48' S, 159°06' E) to GV7 (70°41' S, 158°51' E) analyzed by inductively coupled plasma-sector field mass spectrometry (ICP-SFMS) at a resolution of 5 cm. We find a distinct seasonality of the bromine enrichment signal in most of the cores, with maxima during the austral spring. Iodine shows average concentrations of 0.04 ppb with little variability. No distinct seasonality is found for iodine and sodium. The transect reveals homogeneous air-to-snow fluxes for the three chemical species along the transect due to competing effects of air masses originating from the Ross Sea and the Southern Ocean.

  12. Effects of charge and surface ligand properties of nanoparticles on oxidative stress and gene expression within the gut of Daphnia magna

    Energy Technology Data Exchange (ETDEWEB)

    Dominguez, Gustavo A.; Lohse, Samuel E.; Torelli, Marco; Murphy, Catherine; Hamers, Robert J.; Orr, Galya; Klaper, Rebecca D.

    2015-05-01

    Concern has been raised regarding the current and future release of engineered nanomaterials into aquatic environments from industry and other sources. However, not all nanomaterials may cause an environ-mental impact and identifying which nanomaterials may be of greatest concern has been difficult. It is thought that the surface groups of a functionalized nanoparticles (NPs) may play a significant role in determining their interactions with aquatic organisms, but the way in which surface properties of NPs impact their toxicity in whole organisms has been minimally explored. A major point of interaction of NPs with aquatic organisms is in the gastrointestinal tract as they ingest particulates from the water column or from the sediment. The main goal of this study was to use model gold NP (AuNPs) to evaluate the potential effects of the different surfaces groups on NPs on the gut of an aquatic model organism, Daphnia magna. In this study, we exposed daphnids to a range of AuNPs concentrations and assessed the impact of AuNP exposure in the daphnid gut by measuring reactive oxygen species (ROS) production and expression of genes associated with oxidative stress and general cellular stress: glutathione S-transferase(gst), catalase (cat), heat shock protein 70 (hsp70), and metallothionein1 (mt1). We found ROS formation and gene expression were impacted by both charge and the specific surface ligand used. We detected some degree of ROS production in all NP exposures, but positively charged AuNPs induced a greater ROS response. Similarly, we observed that, compared to controls, both positively charged AuNPs and only one negatively AuNP impacted expression of genes associated with cellular stress. Finally, ligand-AuNP exposures showed a different toxicity and gene expression profile than the ligand alone, indicating a NP specific effect.

  13. Short- and medium-chain fatty acids enhance the cell surface expression and transport capacity of the bile salt export pump (BSEP/ABCB11).

    Science.gov (United States)

    Kato, Takuya; Hayashi, Hisamitsu; Sugiyama, Yuichi

    2010-09-01

    The reduced expression of the bile salt export pump (BSEP/ABCB11) at the canalicular membrane is associated with cholestasis-induced hepatotoxicity due to the accumulation of bile acids in hepatocytes. We previously reported that 4-phenylbutyrate (4PBA), an approved drug for urea cycle disorders, is a promising agent for intrahepatic cholestasis because it increases both the cell surface expression and the transport capacity of BSEP. In the present study, we searched for effective compounds other than 4PBA by focusing on short- and medium-chain fatty acids, which have similar characteristics to 4PBA such as their low-molecular-weight and a carboxyl group. In transcellular transport studies using Madin-Darby canine kidney (MDCK) II cells, all short- and medium-chain fatty acids tested except for formate, acetate, and hexanoic acid showed more potent effects on wild type (WT) BSEP-mediated [3H]taurocholate transport than did 4PBA. The increase in WT BSEP transport with butyrate and octanoic acid treatment correlated with an increase in its expression at the cell surface. Two PFIC2-type variants, E297G and D482G BSEP, were similarly affected with both compounds treatment. The prolonged half-life of cell surface-resident WT BSEP was responsible for this increased octanoic acid-stimulated transport, but not for that of butyrate. In conclusion, short- and medium-chain fatty acids have potent effects on the increase in WT and PFIC2-type BSEP-mediated transport in MDCK II cells. Although both short- and medium-chain fatty acids enhance the transport capacity of WT and PFIC2-type BSEP by inducing those expressions at the cell surface, the underlying mechanism seems to differ between fatty acids. 2010 Elsevier B.V. All rights reserved.

  14. A single point in protein trafficking by Plasmodium falciparum determines the expression of major antigens on the surface of infected erythrocytes targeted by human antibodies.

    Science.gov (United States)

    Chan, Jo-Anne; Howell, Katherine B; Langer, Christine; Maier, Alexander G; Hasang, Wina; Rogerson, Stephen J; Petter, Michaela; Chesson, Joanne; Stanisic, Danielle I; Duffy, Michael F; Cooke, Brian M; Siba, Peter M; Mueller, Ivo; Bull, Peter C; Marsh, Kevin; Fowkes, Freya J I; Beeson, James G

    2016-11-01

    Antibodies to blood-stage antigens of Plasmodium falciparum play a pivotal role in human immunity to malaria. During parasite development, multiple proteins are trafficked from the intracellular parasite to the surface of P. falciparum-infected erythrocytes (IEs). However, the relative importance of different proteins as targets of acquired antibodies, and key pathways involved in trafficking major antigens remain to be clearly defined. We quantified antibodies to surface antigens among children, adults, and pregnant women from different malaria-exposed regions. We quantified the importance of antigens as antibody targets using genetically engineered P. falciparum with modified surface antigen expression. Genetic deletion of the trafficking protein skeleton-binding protein-1 (SBP1), which is involved in trafficking the surface antigen PfEMP1, led to a dramatic reduction in antibody recognition of IEs and the ability of human antibodies to promote opsonic phagocytosis of IEs, a key mechanism of parasite clearance. The great majority of antibody epitopes on the IE surface were SBP1-dependent. This was demonstrated using parasite isolates with different genetic or phenotypic backgrounds, and among antibodies from children, adults, and pregnant women in different populations. Comparisons of antibody reactivity to parasite isolates with SBP1 deletion or inhibited PfEMP1 expression suggest that PfEMP1 is the dominant target of acquired human antibodies, and that other P. falciparum IE surface proteins are minor targets. These results establish SBP1 as part of a critical pathway for the trafficking of major surface antigens targeted by human immunity, and have key implications for vaccine development, and quantifying immunity in populations.

  15. A monoclonal antibody recognizes undifferentiation-specific carbohydrate moieties expressed on cell surface of the human dental pulp cells.

    Science.gov (United States)

    Kang, Kyung-Jung; Ko, Seon-Yle; Ryu, Chun-Jeih; Jang, Young-Joo

    2017-05-01

    Human dental pulp cells are obtained from dental pulp tissue, and have the ability to form dentin and a pulp-like complex. Although adult stem cells have been identified from the primary culture by using specific cell surface markers, the identity of surface markers for the purification of stem cells within the dental pulp population are still unclear. Previously, we had constructed monoclonal antibodies against the undifferentiated cell-specific surface markers of human dental pulp cells (hDPCs) by performing decoy immunization. Among them, a monoclonal antibody against the cell surface antigen of the undifferentiated hDPCs (named UPSA-1) was purified and its heavy and light chain consensus regions were analyzed. The cell surface binding affinity of UPSA-1 mAb on the undifferentiated hDPCs was stronger than that on the differentiated cells. When tunicamycin was applied to hDPSCs during culture, the cell surface binding affinity of the antibody was dramatically decreased, and dentinogenic differentiation was reduced. The purified UPSA-1 antigen band resulting from immunoprecipitation disappeared or shifted down on the SDS-PAGE by deglycosylation. These data suggested that glycosylation on the cell surface might be a marker of an undifferentiated state, and that UPSA-1 mAb might be useful for identifying the carbohydrate moiety on the cell surface of undifferentiated pulp cells. Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.

  16. Microarray expression in peri-implant tissue next to different titanium implant surfaces predicts clinical outcomes: a split-mouth study.

    Science.gov (United States)

    Menini, M; Dellepiane, E; Baldi, D; Longobardi, M G; Pera, P; Izzotti, A

    2017-09-01

    This split-mouth study evaluated miRNA expression of tissues around implants with different surface treatments. Each patient of the sample (five men and five women) received two implants (one control and one test) into an edentulous quadrant to support fixed partial dentures. The control implants (Osseotite) had a dual acid-etched (DAE) surface in the apical portion and a machined coronal part, test implants (Full Osseotite, FOSS) were completely DAE. Machined healing abutments were placed on control implants and DAE abutments on test ones. All implants were assigned codes for blinding. Standardized periapical radiographs were taken at baseline, 2 and 6 months, and 1 year after surgery. Plaque index (PI), bleeding on probing (BOP), and probing depth (PD) were recorded at 3 and 6 weeks, and 2, 3, 6, and 12 months post-implant placement. After 3 months, a mini-invasive sample of soft tissue was collected from seven patients (four women and three men) for miRNA microarray analysis. Control implants showed greater bone resorption (BR) and lower PI: this was not statistically significant. No statistically significant differences in BOP and PD appeared. miRNA modulated by implant surfaces as well as by other clinical conditions has been identified. miRNA microarray analysis revealed that: (i) implant sites with low PI and absence of BOP had a miRNA expression profile similar to those with plaque and absence of BOP; sites with high PI and high BOP had a different profile. (ii) Implant sites with BOP presented similar profiles independently from implant surface. (iii) Implant sites with high PI and normal BR differed from others for miRNA expression profile. (iv) Implant sites with normal BR despite high BOP differed from others. This profile resembled that of FOSS implants. (v) Implant surface affected BR; groups having similar BR clusterized differently according to the implant type. DAE surfaces induced lower BR and more plaque accumulation: This did not affect the

  17. Simulation of surface temperature and ice cover of large northern lakes with 1-D models: a comparison with MODIS satellite data and in situ measurements

    Directory of Open Access Journals (Sweden)

    H. Kheyrollah Pour

    2012-03-01

    Full Text Available Lake surface temperature (LST and ice phenology were simulated for various points differing in depth on Great Slave Lake and Great Bear Lake, two large lakes located in the Mackenzie River Basin in Canada's Northwest Territories, using the 1-D Freshwater Lake model (FLake and the Canadian Lake Ice Model (CLIMo over the 2002–2010 period, forced with data from three weather stations (Yellowknife, Hay River and Deline. LST model results were compared to those derived from the Moderate Resolution Imaging Spectroradiometer (MODIS aboard the Earth Observing System Terra and Aqua satellite platforms. Simulated ice thickness and freeze-up/break-up dates were also compared to in situ observations. Both models showed a good agreement with daily average MODIS LSTs on an annual basis (0.935  ≤  relative index of agreement  ≤  0.984 and 0.94  ≤  mean bias error  ≤  4.83. The absence of consideration of snow on lake ice in FLake was found to have a large impact on estimated ice thicknesses (25 cm thicker on average by the end of winter compared to in situ measurements; 9 cm thicker for CLIMo and break-up dates (6 d earlier in comparison with in situ measurements; 3 d later for CLIMo. The overall agreement between the two models and MODIS LST products during both the open water and ice seasons was good. Remotely sensed data are a promising data source for assimilation into numerical weather prediction models, as they provide the spatial coverage that is not captured by in situ data.

  18. Low Constitutive Cell Surface Expression of HLA-B Is Caused by a Posttranslational Mechanism Involving Glu180 and Arg239

    DEFF Research Database (Denmark)

    Dellgren, Christoffer; Ekwelum, Vanessa A. C.; Ormhøj, Maria

    2016-01-01

    most of the variability of expression between common HLA-A and -B allomorphs. In this study, we present a fine mapping of the structural determinants that allow such variability by exchanging key amino acids located within the C-terminal part of the α2 domain and the α3 domain of HLA-A2 and -B8......, including Glu/Asp at position 177, Gln/Glu at position 180, Gly/Arg at position 239, and Pro/Ser at position 280. We found that the HLA-A2 and -B8 expression profiles could be interconverted to a large extent by mutual exchange of Gln/Glu at position 180 or by Gly/Arg at position 239. The presence of Gln......(180) and Gly(239), as in HLA-A2, led to higher cell surface expression levels when compared with the presence of Glu(180) and Arg(239), as in HLA-B8. This indicates that the amino acids at positions 180 and 239 determine the level of cell surface expression of common HLA-A and -B allomorphs, probably...

  19. HMME-based PDT restores expression and function of transporter associated with antigen processing 1 (TAP1) and surface presentation of MHC class I antigen in human glioma.

    Science.gov (United States)

    Zhang, Shan-Yi; Li, Jun-Liang; Xu, Xin-Ke; Zheng, Mei-Guang; Wen, Cheng-Cai; Li, Fang-Cheng

    2011-11-01

    Numerous studies have established that photodynamic therapy (PDT) can trigger tumor-specific immunity and cancer cell immunogenicity, both of which play a critical role in the long-term control of oncogenesis; however, the underlying mechanisms are largely unexplained. Deficiency of the transporter associated with antigen processing 1 (TAP1) has been observed in a variety of tumors, and the question has been raised whether the restoration of TAP1 could facilitate the activation of antitumor immunity. To elucidate the mechanisms underlying PDT-induced immunopotentiation, we examined the hypothesis that upregulating TAP1 via PDT may contribute to enhancement of antitumor immunity and cancer cell immunogenicity. In this study, we investigated the effects of PDT on the expression and function of TAP1 in glioma cells. We found that HMME-based PDT restored TAP1 expression in a rapid and transient manner. Furthermore, the newly synthesized TAP1 protein was capable of potentiating the activity of transporting antigen peptides. As a result, restoration of the expression and function of TAP1 translated into augmenting the presentation of surface MHC class I molecules. Overall, our data indicate that PDT enables glioma cells to recover both the expression of functional TAP1 and the presentation of surface MHC class I antigens, which are processes that may enhance antitumor immunity after PDT. These findings may have implications for PDT and provide new insights into the mechanisms underlying PDT-induced immunopotentiation.

  20. Histone Deacetylase Inhibitors Restore Cell Surface Expression of the Coxsackie Adenovirus Receptor and Enhance CMV Promoter Activity in Castration-Resistant Prostate Cancer Cells

    Directory of Open Access Journals (Sweden)

    Laura Kasman

    2012-01-01

    Full Text Available Adenoviral gene therapy using the death receptor ligand TRAIL as the therapeutic transgene can be safely administered via intraprostatic injection but has not been evaluated for efficacy in patients. Here we investigated the efficacy of adenoviral TRAIL gene therapy in a model of castration resistant prostate cancer and found that intratumoral injections can significantly delay tumor growth but cannot eliminate established lesions. We hypothesized that an underlying cause is inefficient adenoviral delivery. Using the LNCaP progression model of prostate cancer we show that surface CAR expression decreases with increasing tumorigenicity and that castration resistant C4-2b cells were more difficult to transduce with adenovirus than castration sensitive LNCaP cells. Many genes, including CAR, are epigenetically silenced during transformation but a new class of chemotherapeutic agents, known as histone deacetylase inhibitors (HDACi, can reverse this process. We demonstrate that HDACi restore CAR expression and infectivity in C4-2b cells and enhance caspase activation in response to infection with a TRAIL adenovirus. We also show that in cells with high surface CAR expression, HDACi further enhance transgene expression from the CMV promoter. Thus HDACi have multiple beneficial effects, which may enhance not only viral but also non-viral gene therapy of castration resistant prostate cancer.

  1. Nonstructural Protein 4 of Porcine Reproductive and Respiratory Syndrome Virus Modulates Cell Surface Swine Leukocyte Antigen Class I Expression by Downregulating β2-Microglobulin Transcription.

    Science.gov (United States)

    Qi, Pengfei; Liu, Ke; Wei, Jianchao; Li, Yuming; Li, Beibei; Shao, Donghua; Wu, Zhuanchang; Shi, Yuanyuan; Tong, Guangzhi; Qiu, Yafeng; Ma, Zhiyong

    2017-03-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) is the causative agent of PRRS, which has important impacts on the pig industry. PRRSV infection results in disruption of the swine leukocyte antigen class I (SLA-I) antigen presentation pathway. In this study, highly pathogenic PRRSV (HP-PRRSV) infection inhibited transcription of the β2-microglobulin (β2M) gene ( B2M ) and reduced cellular levels of β2M, which forms a heterotrimeric complex with the SLA-I heavy chain and a variable peptide and plays a critical role in SLA-I antigen presentation. HP-PRRSV nonstructural protein 4 (Nsp4) was involved in the downregulation of β2M expression. Exogenous expression of Nsp4 downregulated β2M expression at both the mRNA and the protein level and reduced SLA-I expression on the cell surface. Nsp4 bound to the porcine B2M promoter and inhibited its transcriptional activity. Domain III of Nsp4 and the enhancer PAM element of the porcine B2M promoter were identified as essential for the interaction between Nsp4 and B2M These findings demonstrate a novel mechanism whereby HP-PRRSV may modulate the SLA-I antigen presentation pathway and provide new insights into the functions of HP-PRRSV Nsp4. IMPORTANCE PRRSV modulates the host response by disrupting the SLA-I antigen presentation pathway. We show that HP-PRRSV downregulates SLA-I expression on the cell surface via transcriptional inhibition of B2M expression by viral Nsp4. The interaction between domain III of Nsp4 and the enhancer PAM element of the porcine B2M promoter is essential for inhibiting B2M transcription. These observations reveal a novel mechanism whereby HP-PRRSV may modulate SLA-I antigen presentation and provide new insights into the functions of viral Nsp4. Copyright © 2017 American Society for Microbiology.

  2. The PPE domain of PPE17 is responsible for its surface localization and can be used to express heterologous proteins on the mycobacterial surface.

    Directory of Open Access Journals (Sweden)

    Valentina Donà

    Full Text Available PPE represent a peculiar family of mycobacterial proteins characterized by a 180 aminoacids conserved N-terminal domain. Several PPE genes are co-transcribed with a gene encoding for a protein belonging to another family of mycobacterial specific proteins named PE. Only one PE-PPE couple has been extensively characterized so far (PE25-PPE41 and it was shown that these two proteins form a heterodimer and that this interaction is essential for PPE41 stability and translocation through the mycobacterial cell wall. In this study we characterize the PE11-PPE17 couple. In contrast with what was found for PE25-PPE41, we show that PPE17 is not secreted but surface exposed. Moreover, we demonstrate that the presence of PE11 is not necessary for PPE17 stability or for its localization on the mycobacterial surface. Finally, we show that the PPE domain of PPE17 targets the mycobacterial cell wall and that this domain can be used as a fusion partner to expose heterologous proteins on the mycobacterial surface.

  3. Water pollution abatement programme. The Czech Republic. Project 4.2. Assessing critical loads of acidity to surface waters in the Czech Republic. Critical loads of acidity to surface waters, north-eastern Bohemia and northern Moravia, The Czech Republic

    Energy Technology Data Exchange (ETDEWEB)

    Lien, L.; Raclavsky, K.; Raclavska, H.; Matysek, D.; Hovind, H.

    1996-01-01

    This report discusses estimates of critical loads of acidity to surface waters and their exceedances, for north-eastern Bohemia and Moravia in The Czech Republic. The survey covers 13 400 km{sup 2}, or 17% of the area of the country. Varying critical loads were observed within the examined region. 19% of the examined area showed exceedance of critical load and another 11% was close to exceedance. The survey should continue in Bohemia. 24 refs., 20 figs., 4 tabs.

  4. Detection of vitellogenin and zona radiata protein expressions in surface mucus of immature juvenile Atlantic salmon (Salmo salar) exposed to waterborne nonylphenol

    International Nuclear Information System (INIS)

    Meucci, Valentina; Arukwe, Augustine

    2005-01-01

    Induction of blood plasma vitellogenin (Vtg) and zona radiata proteins (Zr-proteins) in male and juvenile of oviparous vertebrates was proposed and shown to be sensitive biomarkers for exposure to estrogen mimic. The time- and dose-dependent expression of Vtg and Zr-proteins in nonylphenol (NP) exposed juvenile Atlantic salmon (Salmo salar) is reported. Fish were exposed continuously to waterborne nonylphenol at 5, 15 and 50 μg/L. Blood and surface mucus samples were collected after 3 and 7 days post-exposure. Nonylphenol-induced plasma and surface mucus levels of Vtg and Zr-protein were analysed using immunochemical methods (Western blot and enzyme-linked immunosorbent assay; ELISA). Both Vtg and Zr-protein levels in plasma and surface mucus showed similar and parallel nonylphenol-induced expression patterns after waterborne nonylphenol exposure and in a concentration- and time-dependent manner. Zr-proteins were significantly induced at the lowest concentration of nonylphenol after 3 and 7 days of exposure both in plasma and in surface mucus. We conclude that the detection of Vtg and Zr-proteins directly in the surface mucus of fish, and the correlation of these values with plasma protein biomarker values in xenoestrogen-treated fish represents a sensitive non-invasive system for the detection of these known endocrine disruptor biomarkers. The demonstration of detectable Vtg and Zr-protein levels from surface mucus is a potential biomarker for estrogenic compounds, and their presence should be considered as an improvement in the methods for detection of endocrine disrupting chemicals (EDCs) and related pollutants in the environment

  5. Cultured Pearl Surface Quality Profiling by the Shell Matrix Protein Gene Expression in the Biomineralised Pearl Sac Tissue of Pinctada margaritifera.

    Science.gov (United States)

    Blay, Carole; Planes, Serge; Ky, Chin-Long

    2018-04-16

    Nucleated pearls are produced by molluscs of the Pinctada genus through the biomineralisation activity of the pearl sac tissue within the recipient oyster. The pearl sac originates from graft tissue taken from the donor oyster mantle and its functioning is crucial in determining key factors that impact pearl quality surface characteristics. The specific role of related gene regulation during gem biogenesis was unknown, so we analysed the expression profiles of eight genes encoding nacreous (PIF, MSI60, PERL1) or prismatic (SHEM5, PRISM, ASP, SHEM9) shell matrix proteins or both (CALC1) in the pearl sac (N = 211) of Pinctada margaritifera during pearl biogenesis. The pearls and pearl sacs analysed were from a uniform experimental graft with sequential harvests at 3, 6 and 9 months post-grafting. Quality traits of the corresponding pearls were recorded: surface defects, surface deposits and overall quality grade. Results showed that (1) the first 3 months of culture seem crucial for pearl quality surface determination and (2) all the genes (SHEM5, PRISM, ASP, SHEM9) encoding proteins related to calcite layer formation were over-expressed in the pearl sacs that produced low pearl surface quality. Multivariate regression tree building clearly identified three genes implicated in pearl surface quality, SHEM9, ASP and PIF. SHEM9 and ASP were clearly implicated in low pearl quality, whereas PIF was implicated in high quality. Results could be used as biomarkers for genetic improvement of P. margaritifera pearl quality and constitute a novel perspective to understanding the molecular mechanism of pearl formation.

  6. Detection of vitellogenin and zona radiata protein expressions in surface mucus of immature juvenile Atlantic salmon (Salmo salar) exposed to waterborne nonylphenol

    Energy Technology Data Exchange (ETDEWEB)

    Meucci, Valentina [Department of Veterinary Clinics, Section of Pharmacology and Toxicology, University of Pisa, V. le delle Piagge 2, 56124 Pisa (Italy); Arukwe, Augustine [Department of Biology, Norwegian University of Science and Technology (NTNU), Hoyskoleringen 5, 7491 Trondheim (Norway)]. E-mail: arukwe@bio.ntnu.no

    2005-06-01

    Induction of blood plasma vitellogenin (Vtg) and zona radiata proteins (Zr-proteins) in male and juvenile of oviparous vertebrates was proposed and shown to be sensitive biomarkers for exposure to estrogen mimic. The time- and dose-dependent expression of Vtg and Zr-proteins in nonylphenol (NP) exposed juvenile Atlantic salmon (Salmo salar) is reported. Fish were exposed continuously to waterborne nonylphenol at 5, 15 and 50 {mu}g/L. Blood and surface mucus samples were collected after 3 and 7 days post-exposure. Nonylphenol-induced plasma and surface mucus levels of Vtg and Zr-protein were analysed using immunochemical methods (Western blot and enzyme-linked immunosorbent assay; ELISA). Both Vtg and Zr-protein levels in plasma and surface mucus showed similar and parallel nonylphenol-induced expression patterns after waterborne nonylphenol exposure and in a concentration- and time-dependent manner. Zr-proteins were significantly induced at the lowest concentration of nonylphenol after 3 and 7 days of exposure both in plasma and in surface mucus. We conclude that the detection of Vtg and Zr-proteins directly in the surface mucus of fish, and the correlation of these values with plasma protein biomarker values in xenoestrogen-treated fish represents a sensitive non-invasive system for the detection of these known endocrine disruptor biomarkers. The demonstration of detectable Vtg and Zr-protein levels from surface mucus is a potential biomarker for estrogenic compounds, and their presence should be considered as an improvement in the methods for detection of endocrine disrupting chemicals (EDCs) and related pollutants in the environment.

  7. Prostaglandin E2 Receptor Expression by Osteoblasts is Modulated by Implant Surface Roughness and Prostaglandin E2

    Science.gov (United States)

    2006-05-01

    the surface of a load- carrying implant" ( Branemark et al. 1977). The current American Academy of Periodontology definition (2001) describes...inhabit and produce bone very close to its surface, allowing for the necessary proximity for integration of dental implants ( Branemark et al. 1969). The...component of titanium’s biocompatibility and suitability as a restorative metal ( Branemark et al. 1977). In titanium-aluminum-vanadium alloy (Ti-6A1-4V), it

  8. Estimating the Mean Annual Surface Air Temperature at Armagh Observatory, Northern Ireland, and the Global Land-Ocean Temperature Index for Sunspot Cycle 24, the Current Ongoing Sunspot Cycle

    Science.gov (United States)

    Wilson, Robert M.

    2013-01-01

    As noted by Gray et al., Sir William Herschel was the first to suggest a possible close connection between the Sun and the Earth’s climate. The Sun, being the source of energy that impacts and drives the Earth’s climate system, displays a variety of changes over both short and long term time scales, the most obvious examples being the somewhat regular waxing and waning of sunspots with time (i.e., the sunspot cycle (SC)), first described by Samuel Heinrich Schwabe, a German apothecary and amateur astronomer who observed the Sun from Dessau, Germany, and the now well established variation of the Sun’s irradiance over the SC. Other factors related to the SC have been linked to changes in climate as well. Some of these other factors include the role of cosmic rays and the solar wind (i.e., the geomagnetic cycle) on climate, as well as the apparent close association between trends in global and northern hemispheric temperature and the length of the SC, although some investigators have described the inferred association between climate and, in particular, SC length as now being weak. More recently, Solheim et al. have reported on the relation between SC length and the average temperature in the same and immediately following SC for a number of meteorological stations in Norway and in the North Atlantic region. They noted that while they found no significant trend (correlation) between SC length and the average temperature when measured for the same cycle, in contrast, they found a significant negative trend when SC length was compared with the following cycle’s average temperature. From this observation, they suggested that average northern hemispheric temperature during the present ongoing SC (SC24) will be lower by about 0.9 °C than was seen in SC23 (spanning 1996–2007, based on yearly averages of sunspot number (SSN), and onset for SC24 occurring in 2008). The purpose of this Technical Publication (TP) is to examine the annual variations of the Armagh

  9. Differential Susceptibility of Bighorn Sheep (Ovis canadensis) and Domestic Sheep (Ovis aries) Neutrophils to Mannheimia haemolytica Leukotoxin is not due to Differential Expression of Cell Surface CD18.

    Science.gov (United States)

    Dassanayake, Rohana P; Shanthalingam, Sudarvili; Liu, Weiguo; Casas, Eduardo; Srikumaran, Subramaniam

    2017-07-01

    Bighornsheep ( Ovis canadensis ) are more susceptible to pneumonia caused by Mannheimia haemolytica than are domestic sheep ( Ovis aries ). Leukotoxin produced by M. haemolytica is the principal virulence factor involved in pneumonia pathogenesis. Although leukotoxin is cytolytic to all subsets of ruminant leukocytes, neutrophils are the most susceptible subset. Bighorn sheep neutrophils are four- to eightfold more susceptible to leukotoxin-induced cytolysis than are domestic sheep neutrophils. We hypothesized that the higher susceptibility of bighorn sheep neutrophils, in comparison to domestic sheep neutrophils, is due to higher expression of CD18, the receptor for leukotoxin on leukocytes. Our objective was to quantify CD18 expression on neutrophils of bighorn sheep and domestic sheep. Cell-surface CD18 expression on bighorn sheep and domestic sheep neutrophils was measured as antibody binding capacity of cells by flow cytometric analysis with two fluorochrome-conjugated anti-CD18 monoclonal antibodies (BAQ30A and HUH82A) and microspheres. Contrary to our expectations, CD18 expression was higher (Psheep neutrophils in comparison to bighorn sheep neutrophils. These findings suggest that the higher in vitro susceptibility to leukotoxin of bighorn sheep neutrophils compared to domestic sheep neutrophils is not due to higher expression of the leukotoxin receptor CD18 on bighorn sheep neutrophils.

  10. Constitutive expression and characterization of a surface SRS (NcSRS67) protein of Neospora caninum with no orthologue in Toxoplasma gondii.

    Science.gov (United States)

    Bezerra, Marcos Alexandre; Pereira, Luiz Miguel; Bononi, Aline; Biella, Carla Agostino; Baroni, Luciana; Pollo-Oliveira, Leticia; Yatsuda, Ana Patrícia

    2017-04-01

    Neospora caninum is a parasite of the Apicomplexa phylum responsible for abortion and losses of fertility in cattle. As part of its intracellular cycle, the first interaction of the parasite with the target cell is performed with the surface proteins known as the SRS superfamily (Surface Antigen Glycoprotein - Related Sequences). SAG related or SRS proteins have been a target of intense research due to its immunodominant pattern, exhibiting potential as diagnostic and/or vaccine candidates. The aim of this study was the cloning, expression and characterization of the gene NcSRS67 of N. caninum using a novel designed plasmid. The coding sequence of NcSRS67 (without the signal peptide and the GPI anchor) was cloned and expressed constitutively instead of the ccdB system of pCR-Blunt II-TOPO. The protein was purified in a nickel sepharose column and identified by mass spectrometry (MS/MS). The constitutive expression did not affect the final bacterial growth, with a similar OD 600nm compared to the non-transformed strains. The recombinant NcSRS67 was over expressed and the native form was detected by the anti-rNcSRS67 serum on 1D western blot as a single band of approximately 38kDa as predicted. On an in vitro assay, the inhibitory effect of the polyclonal antiserum anti-rNcSRS67 was nearly 20% on adhesion/invasion of host cells. The NcSRS67 native protein was localised on part of the surface of N. caninum tachyzoite when compared to the nucleus by confocal immunofluorescence. Copyright © 2017 Elsevier B.V. All rights reserved.

  11. Recent climate changes in the northern hemisphere

    International Nuclear Information System (INIS)

    Trenberth, K.E.

    1990-01-01

    The consistency of analyzed changes in surface wind stress, sea level pressures and surface temperatures between 1980-86 and previous periods indicates the reality of statistically significant and substantial climate changes in the Northern Hemisphere, especially over the North Pacific, on decadal time scales. Cooling in North Pacific sea surface temperatures and warming along the west coast of North America and Alaska are ascribed mainly to the changes in thermal advection associated with a deeper and more extensive Aleutian Low

  12. A PDZ-Like Motif in the Biliary Transporter ABCB4 Interacts with the Scaffold Protein EBP50 and Regulates ABCB4 Cell Surface Expression.

    Directory of Open Access Journals (Sweden)

    Quitterie Venot

    Full Text Available ABCB4/MDR3, a member of the ABC superfamily, is an ATP-dependent phosphatidylcholine translocator expressed at the canalicular membrane of hepatocytes. Defects in the ABCB4 gene are associated with rare biliary diseases. It is essential to understand the mechanisms of its canalicular membrane expression in particular for the development of new therapies. The stability of several ABC transporters is regulated through their binding to PDZ (PSD95/DglA/ZO-1 domain-containing proteins. ABCB4 protein ends by the sequence glutamine-asparagine-leucine (QNL, which shows some similarity to PDZ-binding motifs. The aim of our study was to assess the potential role of the QNL motif on the surface expression of ABCB4 and to determine if PDZ domain-containing proteins are involved. We found that truncation of the QNL motif decreased the stability of ABCB4 in HepG2-transfected cells. The deleted mutant ABCB4-ΔQNL also displayed accelerated endocytosis. EBP50, a PDZ protein highly expressed in the liver, strongly colocalized and coimmunoprecipitated with ABCB4, and this interaction required the QNL motif. Down-regulation of EBP50 by siRNA or by expression of an EBP50 dominant-negative mutant caused a significant decrease in the level of ABCB4 protein expression, and in the amount of ABCB4 localized at the canalicular membrane. Interaction of ABCB4 with EBP50 through its PDZ-like motif plays a critical role in the regulation of ABCB4 expression and stability at the canalicular plasma membrane.

  13. Hydroxy decenoic acid down regulates gtfB and gtfC expression and prevents Streptococcus mutans adherence to the cell surfaces

    Directory of Open Access Journals (Sweden)

    Yousefi Behnam

    2012-07-01

    Full Text Available Abstract Background 10-Hydroxy-2-decenoic acid, an unsaturated fatty acid is the most active and unique component to the royal jelly that has antimicrobial properties. Streptococcus mutans is associated with pathogenesis of oral cavity, gingivoperiodontal diseases and bacteremia following dental manipulations. In the oral cavity, S. mutans colonize the soft tissues including tongue, palate, and buccal mucosa. When considering the role of supragingival dental plaque in caries, the proportion of acid producing bacteria (particularly S. mutans, has direct relevance to the pathogenicity of the plaque. The genes that encode glucosyltransferases (gtfs especially gtfB and gtfC are important in S. mutans colonization and pathogenesis. This study investigated the hydroxy-decenoic acid (HDA effects on gtfB and gtfC expression and S. mutans adherence to cells surfaces. Methods Streptococcus mutans was treated by different concentrations of HPLC purified HDA supplied by Iran Beekeeping and Veterinary Association. Real time RT-PCR and western blot assays were conducted to evaluate gtfB and gtfC genes transcription and translation before and after HDA treatment. The bacterial attachment to the cell surfaces was evaluated microscopically. Results 500 μg ml-1 of HDA inhibited gtfB and gtfC mRNA transcription and its expression. The same concentration of HDA decreased 60% the adherence of S. mutans to the surface of P19 cells. Conclusion Hydroxy-decenoic acid prevents gtfB and gtfC expression efficiently in the bactericide sub-concentrations and it could effectively reduce S. mutans adherence to the cell surfaces. In the future, therapeutic approaches to affecting S. mutans could be selective and it’s not necessary to put down the oral flora completely.

  14. Mutations within the Pathogenic Region of Herpes Simplex Virus 1 gK Signal Sequences Alter Cell Surface Expression and Neurovirulence

    Science.gov (United States)

    Matundan, Harry H.; Mott, Kevin R.; Akhtar, Aslam Abbasi; Breunig, Joshua J.

    2014-01-01

    ABSTRACT To investigate the role of the signal sequences of herpes simplex virus 1 (HSV-1) gK on virus replication and viral pathogenesis, we constructed recombinant viruses with or without mutations within the signal sequences of gK. These recombinant viruses expressed two additional copies of the mutated (MgK) or native (NgK) form of the gK gene in place of the latency-associated transcript with a myc epitope tag to facilitate detection at their 3′ ends. The replication of MgK virus was similar to that of NgK both in vitro and in vivo, as well as in the trigeminal ganglia (TG) of latently infected mice. The levels of gB and gK transcripts in the corneas, TG, and brains of infected mice on days 3 and 5 postinfection were markedly virus and time dependent, as well as tissue specific. Mutation in the signal sequence of gK in MgK virus blocked cell surface expression of gK-myc in rabbit skin cells, increased 50% lethal dose, and decreased corneal scarring in ocularly infected mice compared to the NgK or revertant (RgK) virus. MgK and NgK viruses, and not the RgK virus, showed a reduced extent of explant reactivation at the lower dose of ocular infection but not at the higher dose. However, the time of reactivation was not affected by overexpression of the different forms of gK. Taken together, these results strongly suggest that the 8mer peptide (ITAYGLVL) within the signal sequence of gK promotes cell surface expression of gK in infected cells and ocular pathogenesis in infected mice. IMPORTANCE In this study, we show for the first time that mutations within the signal sequence of gK blocked cell surface expression of inserted recombinant gK in vitro. Furthermore, this blockage in cell surface expression was correlated with higher 50% lethal dose and less corneal scarring in vivo. Thus, these studies point to a key role for the 8mer within the signal sequence of gK in HSV-1-induced pathogenicity. PMID:25505072

  15. Loss of ATP-Sensitive Potassium Channel Surface Expression in Heart Failure Underlies Dysregulation of Action Potential Duration and Myocardial Vulnerability to Injury.

    Directory of Open Access Journals (Sweden)

    Zhan Gao

    Full Text Available The search for new approaches to treatment and prevention of heart failure is a major challenge in medicine. The adenosine triphosphate-sensitive potassium (KATP channel has been long associated with the ability to preserve myocardial function and viability under stress. High surface expression of membrane KATP channels ensures a rapid energy-sparing reduction in action potential duration (APD in response to metabolic challenges, while cellular signaling that reduces surface KATP channel expression blunts APD shortening, thus sacrificing energetic efficiency in exchange for greater cellular calcium entry and increased contractile force. In healthy hearts, calcium/calmodulin-dependent protein kinase II (CaMKII phosphorylates the Kir6.2 KATP channel subunit initiating a cascade responsible for KATP channel endocytosis. Here, activation of CaMKII in a transaortic banding (TAB model of heart failure is coupled with a 35-40% reduction in surface expression of KATP channels compared to hearts from sham-operated mice. Linkage between KATP channel expression and CaMKII is verified in isolated cardiomyocytes in which activation of CaMKII results in downregulation of KATP channel current. Accordingly, shortening of monophasic APD is slowed in response to hypoxia or heart rate acceleration in failing compared to non-failing hearts, a phenomenon previously shown to result in significant increases in oxygen consumption. Even in the absence of coronary artery disease, failing myocardium can be further injured by ischemia due to a mismatch between metabolic supply and demand. Ischemia-reperfusion injury, following ischemic preconditioning, is diminished in hearts with CaMKII inhibition compared to wild-type hearts and this advantage is largely eliminated when myocardial KATP channel expression is absent, supporting that the myocardial protective benefit of CaMKII inhibition in heart failure may be substantially mediated by KATP channels. Recognition of Ca

  16. The extracellular membrane-proximal domain of membrane-bound IgE restricts B cell activation by limiting B cell antigen receptor surface expression.

    Science.gov (United States)

    Vanshylla, Kanika; Opazo, Felipe; Gronke, Konrad; Wienands, Jürgen; Engels, Niklas

    2018-03-01

    Immunoglobulin E (IgE) antibodies are key mediators of allergic reactions. Due to their potentially harmful anaphylactic properties, their production is tightly regulated. The membrane-bound isoform of IgE (mIgE), which is an integral component of the B cell antigen receptor, has been shown to be critical for the regulation of IgE responses in mice. In primate species including humans, mIgE can be expressed in two isoforms that are produced by alternative splicing of the primary ε Ig heavy chain transcript, and differ in the absence or presence of an extracellular membrane-proximal domain (EMPD) consisting of 52 amino acids. However, the function of the EMPD remains unclear. Here, we demonstrate that the EMPD restricts surface expression of mIgE-containing BCRs in human and murine B cells. The EMPD does not interfere with BCR assembly but acts as an autonomous endoplasmic reticulum retention domain. Limited surface expression of EMPD-containing mIgE-BCRs caused impaired activation of intracellular signaling cascades and hence represents a regulatory mechanism that may control the production of potentially anaphylactic IgE antibodies in primate species. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  17. Uropathogenic Escherichia coli Express Type 1 Fimbriae Only in Surface Adherent Populations Under Physiological Growth Conditions

    DEFF Research Database (Denmark)

    Stærk, Kristian; Kolmos, Hans Jørn; Khandige, Surabhi

    2016-01-01

    BACKGROUND:  Most uropathogenic Escherichia coli (UPEC) strains harbor genes encoding adhesive type 1 fimbria (T1F). T1F is a key factor for successful establishment of urinary tract infection. However, UPEC strains typically do not express T1F in the bladder urine, and little is understood about...... its induction in vivo. METHODS:  A flow chamber infection model was used to grow UPEC under conditions simulating distinct infection niches in the bladder. Type 1 fimbriation on isolated UPEC was subsequently determined by yeast cell agglutination and immunofluorescence microscopy, and the results...... were correlated with the ability to adhere to and invade cultured human bladder cells. RESULTS:  Although inactive during planktonic growth in urine, T1F expression occurs when UPEC settles on and infects bladder epithelial cells or colonizes catheters. As a result, UPEC in these sessile populations...

  18. Biosynthesis of intestinal microvillar proteins. Processing of N-linked carbohydrate is not required for surface expression

    DEFF Research Database (Denmark)

    Danielsen, Erik Michael; Cowell, G M

    1986-01-01

    microvillar enzymes, it does not receive post-translational O-linked carbohydrate. Castanospermine suppressed the synthesis of the four enzymes, but did not block their transport to the microvillar membrane, showing that processing of N-linked carbohydrate is not required for microvillar expression....... The proteinase inhibitor leupeptin partially restored the suppressed synthesis, indicating that the majority of the wrongly processed enzymes, probably because of conformational instability, become degraded soon after synthesis rather than being transported to the microvillar membrane....

  19. Subtilisin QK-2: secretory expression in Lactococcus lactis and surface display onto gram-positive enhancer matrix (GEM) particles

    OpenAIRE

    Mao, Ruifeng; Zhou, Kangping; Han, Zhenwei; Wang, Yefu

    2016-01-01

    Background Purified from the supernatant of Bacillus subtilis QK02 culture broth, Subtilisin QK-2 is a type of effective thrombolytic reagent that has great exploitable potential. However, the unbearable flavor that occurs with fermentation and the complicated methods that are required to obtain pure products limit the application of this enzyme. Lactic acid bacteria (LAB)-based delivery vehicles are promising as cheap and safe options for medicinal compounds. The secretory expression and sur...

  20. Intermediate monocytes in ANCA vasculitis: increased surface expression of ANCA autoantigens and IL-1β secretion in response to anti-MPO antibodies.

    LENUS (Irish Health Repository)

    O'Brien, Eóin C

    2015-01-01

    ANCA vasculitis encompasses several autoimmune conditions characterised by destruction of small vessels, inflammation of the respiratory tract and glomerulonephritis. Most patients harbour autoantibodies to myeloperoxidase (MPO) or proteinase 3 (PR3). Clinical and experimental data suggest that pathogenesis is driven by ANCA-mediated activation of neutrophils and monocytes. We investigated a potential role for distinct monocyte subsets. We found that the relative proportion of intermediate monocytes is increased in patients versus control individuals, and both MPO and PR3 are preferentially expressed on these cells. We demonstrate that MPO and PR3 are expressed independently of each other on monocytes and that PR3 is not associated with CD177. MPO expression correlates with that of Fc receptor CD16 on intermediate monocytes. Monocyte subsets respond differently to antibodies directed against MPO and PR3, with anti-MPO but not anti-PR3 leading to increased IL-1β, IL-6 and IL-8 production. In concordance with the observed higher surface expression of MPO on intermediate monocytes, this subset produces the highest quantity of IL-1β in response to anti-MPO stimulation. These data suggest that monocytes, specifically, the intermediate subset, may play a role in ANCA vasculitis, and also indicate that substantial differences exist between the effect of anti-MPO and anti-PR3 antibodies on these cells.

  1. Leukocytes-depleting filters preferentially remove activated leukocytes and reduce the expression of surface adhesion molecules during the simulated extracorporeal circulation of human blood.

    Science.gov (United States)

    Alexiou, Christos; Sheppard, Stuart; Tang, Augustine; Rengarajan, Arvind; Smith, David; Haw, Marcus; Gibbs, Roz

    2006-01-01

    The effect of leukocyte-depleting filters on the total and activated leukocyte counts and the expression of surface adhesion molecules CD11b, CD18, and CD62L during the in vitro extracorporeal circulation of human blood was studied. A 200 ml blood sample was taken from 10 patients undergoing CABG surgery. The blood was circulated for 60 minutes within an experimental extracorporeal circuit. A leukocyte-depleting filter was attached in five circuits (filtered group). In five other circuits, no filter was used (controls). Total leukocyte counts were determined manually. Activated leukocytes were identified using nitroblue tetrazolium staining. The expression of CD11b, CD18, and CD62L was measured with flow cytometry. At 60 minutes, total leukocyte counts were reduced by 49% from the baseline values in the filtered group and 10% in the control group (p filtered group and increased by 116% in the control group (p filtered group, the expression of CD11b, CD18, and CD612L decreased by 60%, 21%, and 79%, respectively, and in the control group it increased by 24%, 6%, and 28% (p filters preferentially remove activated leukocytes and reduce the expression of CD11b, CD18, and CD62L during the in vitro extracorporeal circulation of human blood.

  2. Use of T7 RNA polymerase to direct expression of outer Surface Protein A (OspA) from the Lyme disease Spirochete, Borrelia burgdorferi

    Science.gov (United States)

    Dunn, John J.; Lade, Barbara N.

    1991-01-01

    The OspA gene from a North American strain of the Lyme disease Spirochete, Borrelia burgdorferi, was cloned under the control of transciption and translation signals from bacteriophage T7. Full-length OspA protein, a 273 amino acid (31kD) lipoprotein, is expressed poorly in Escherichia coli and is associated with the insoluble membrane fraction. In contrast, a truncated form of OspA lacking the amino-terminal signal sequence which normally would direct localization of the protein to the outer membrane is expressed at very high levels (less than or equal to 100 mg/liter) and is soluble. The truncated protein was purified to homogeneity and is being tested to see if it will be useful as an immunogen in a vaccine against Lyme disease. Circular dichroism and fluorescence spectroscopy was used to characterize the secondary structure and study conformational changes in the protein. Studies underway with other surface proteins from B burgdorferi and a related spirochete, B. hermsii, which causes relapsing fever, leads us to conclude that a strategy similar to that used to express the truncated OspA can provide a facile method for producing variations of Borrelia lipoproteins which are highly expressed in E. coli and soluble without exposure to detergents.

  3. Separate Developmental Programs for HLA-A and -B Cell Surface Expression during Differentiation from Embryonic Stem Cells to Lymphocytes, Adipocytes and Osteoblasts

    DEFF Research Database (Denmark)

    Sabir, Hardee J; Nehlin, Jan O; Qanie, Diyako

    2013-01-01

    hematopoietic stem cells (hHSC), human mesenchymal stem cells (hMSC) and their fully-differentiated progenies such as lymphocytes, adipocytes and osteoblasts. hESC showed extremely low levels of HLA-A and no -B. In contrast, multipotent hMSC and hHSC generally expressed higher levels of HLA-A and clearly HLA......A major problem of allogeneic stem cell therapy is immunologically mediated graft rejection. HLA class I A, B, and Cw antigens are crucial factors, but little is known of their respective expression on stem cells and their progenies. We have recently shown that locus-specific expression (HLA......-A, but not -B) is seen on some multipotent stem cells, and this raises the question how this is in other stem cells and how it changes during differentiation. In this study, we have used flow cytometry to investigate the cell surface expression of HLA-A and -B on human embryonic stem cells (hESC), human...

  4. Protective role of surface Toll-like receptor 9 expressing neutrophils in local inflammation during systemic inflammatory response syndrome in mice.

    Science.gov (United States)

    Meng, Xiuping; Sun, Wei; Ren, Yunjia; Xiao, Yue; Zhao, Peiyan; Lu, Wenting; Hua, Li; Wang, Luowei; Wang, Liying; Yu, Yongli

    2017-10-01

    Clinically, systemic inflammatory response syndrome (SIRS) occurs after serious trauma or sepsis. In sepsis, neutrophils are the major effector cells responsible for eliminating pathogens. However, the role of neutrophils in development of SIRS, especially in local inflammatory area, is controversial. In this study, we established a SIRS mouse model characterized with cytokine-mediated lethal shock by intraperitoneal injection of oligodexynucleotides containing CpG motifs (CpG ODN) in D-galactosamine (D-GalN) sensitized mice based on our previous work and found that abundant neutrophils were rapidly recruited into the peritoneal cavity, where some neutrophils expressed surface TLR9 (sTLR9), defined as sTLR9 + neutrophils. Along with the progression of SIRS, the expression of sTLR9 in sTLR9 + neutrophils isolated from peritoneal lavage cells (PLCs) was declined in accompany with an increase in the level of the inflammatory cytokine TNFα and a decrease in the level of the anti-inflammatory cytokine IL-10 in Ly6G + PLCs. When using CCT ODN, an oligodeoxynucleotide with CCT repeats, which we have previously shown to be capable of rescuing mice from lethal shock, the expression of sTLR9 on neutrophils was significantly elevated. Adoptive therapy using early recruited neutrophil-rich PLCs containing sTLR9 + neutrophils that express high levels of sTLR9, could rescue mice from SIRS. Overall, the data reveal that the early recruited sTLR9 + neutrophils may, at least in the area of local inflammation, play a protective role during SIRS development and provide a method to rescue the patients with severe SIRS via the up-regulation of sTLR9 levels on the surface of neutrophils or via adoptive therapy with protective sub-populations of neutrophils. Copyright © 2017 Elsevier Ltd. All rights reserved.

  5. Changes of exoskeleton surface roughness and expression of crucial participation genes for chitin formation and digestion in the mud crab (Macrophthalmus japonicus) following the antifouling biocide irgarol.

    Science.gov (United States)

    Park, Kiyun; Nikapitiya, Chamilani; Kim, Won-Seok; Kwak, Tae-Soo; Kwak, Ihn-Sil

    2016-10-01

    Irgarol is a common antifoulant present in coastal sediment. The mud crab Macrophthalmus japonicus is one of the most abundant of the macrobenthos in the costal environment, and its exoskeleton has a protective function against various environmental threats. We evaluated the effects of irgarol toxicity on the exoskeleton of M. japonicus, which is the outer layer facing the environment. We analyzed transcriptional expression of exoskeleton, molting, and proteolysis-related genes in the gill and hepatopancreas of these exposed M. japonicus. In addition, changes in survival and exoskeleton surface characteristics were investigated. In the hepatopancreas, mRNA expression of chitinase 1 (Mj-chi1), chitinase 4 (Mj-chi4), and chitinase 5 (Mj-chi5) increased in M. japonicus exposed to all concentrations of irgarol. Mj-chi1 and Mj-chi4 expressions from 1 to 10μgL(-1) were dose- and time-dependent. Ecdysteroid receptor (Mj-EcR), trypsin (Mj-Tryp), and serine proteinase (Mj-SP) in the hepatopancreas were upregulated in response to different exposure levels of irgarol at day 1, 4, or 7. In contrast, gill Mj-chi5, Mj-Tryp, and Mj-SP exhibited late upregulated responses to 10μgL(-1) irgarol compared to the control at day 7. Mj-chi1 showed early upregulation upon exposure to 10μgL(-1) irgarol and Mj-chi4 showed no changes in transcription in the gill. Gill Mj-EcR presented generally downregulated expression patterns. In addition, decreased survival and change of exoskeleton surface roughness were observed in M. japonicus exposed to the three concentrations of irgarol. These results suggest that exposure to irgarol induces changes in the exoskeleton, molting, and proteolysis metabolism of M. japonicus. Copyright © 2016 Elsevier Inc. All rights reserved.

  6. Microbial Disease Spectrum Linked to a Novel IL-12Rβ1 N-Terminal Signal Peptide Stop-Gain Homozygous Mutation with Paradoxical Receptor Cell-Surface Expression

    Science.gov (United States)

    Louvain de Souza, Thais; de Souza Campos Fernandes, Regina C.; Azevedo da Silva, Juliana; Gomes Alves Júnior, Vladimir; Gomes Coelho, Adelia; Souza Faria, Afonso C.; Moreira Salomão Simão, Nabia M.; Souto Filho, João T.; Deswarte, Caroline; Boisson-Dupuis, Stéphanie; Torgerson, Dara; Casanova, Jean-Laurent; Bustamante, Jacinta; Medina-Acosta, Enrique

    2017-01-01

    Patients with Mendelian Susceptibility to Mycobacterial Diseases (MSMD) exhibit variable vulnerability to infections by mycobacteria and other intramacrophagic bacteria (e.g., Salmonella and Klebsiella) and fungi (e.g., Histoplasma, Candida, Paracoccidioides, Coccidioides, and Cryptococcus). The hallmark of MSMD is the inherited impaired production of interferon gamma (IFN-γ) or the lack of response to it. Mutations in the interleukin (IL)-12 receptor subunit beta 1 (IL12RB1) gene accounts for 38% of cases of MSMD. Most IL12RB1 pathogenic allele mutations, including ten known stop-gain variants, cause IL-12Rβ1 complete deficiency (immunodeficiency-30, IMD30) by knocking out receptor cell-surface expression. IL12RB1 loss-of-function genotypes impair both IL-12 and IL-23 responses. Here, we assess the health effects of a rare, novel IL12RB1 stop-gain homozygous genotype with paradoxical IL-12Rβ1 cell-surface expression. We appraise four MSMD children from three unrelated Brazilian kindreds by clinical consultation, medical records, and genetic and immunologic studies. The clinical spectrum narrowed down to Bacillus Calmette-Guerin (BCG) vaccine-related suppurative adenitis in all patients with one death, and recrudescence in two, histoplasmosis, and recurrence in one patient, extraintestinal salmonellosis in one child, and cutaneous vasculitis in another. In three patients, we established the homozygous Trp7Ter predicted loss-of-function inherited genotype and inferred it from the heterozygote parents of the fourth case. The Trp7Ter mutation maps to the predicted IL-12Rβ1 N-terminal signal peptide sequence. BCG- or phytohemagglutinin-blasts from the three patients have reduced cell-surface expression of IL-12Rβ1 with impaired production of IFN-γ and IL-17A. Screening of 227 unrelated healthy subjects from the same geographic region revealed one heterozygous genotype (allele frequency 0.0022) vs. one in over 841,883 public genome/exomes. We also show that the

  7. Aberrantly glycosylated MUC1 is expressed on the surface of breast cancer cells and a target for antibody-dependent cell-mediated cytotoxicity

    DEFF Research Database (Denmark)

    Lavrsen, Kirstine; Madsen, Caroline B; Rasch, Morten G

    2013-01-01

    not covered by immunological tolerance in MUC1 humanized mice and man. The objective of this study was to determine if mouse antibodies to this Tn-MUC1 epitope induce antibody-dependent cellular cytotoxicity (ADCC) pivotal for their potential use in cancer immunotherapy. Binding affinity of mAb 5E5 directed...... to Tn-MUC1 was investigated using BiaCore. The availability of Tn-MUC1 on the surface of breast cancer cells was evaluated by immunohistochemistry, confocal microscopy, and flow cytometry, followed by in vitro assessment of antibody-dependent cellular cytotoxicity by mAb 5E5. Biacore analysis...... is expressed on the surface of breast cancer cells and a target for antibody-dependent cell-mediated cytotoxicity suggesting that antibodies targeting glycopeptide epitopes on mucins are strong candidates for cancer-specific immunotherapies....

  8. Surface chemistry of gold nanoparticles determines the biocorona composition impacting cellular uptake, toxicity and gene expression profiles in human endothelial cells.

    Science.gov (United States)

    Chandran, Parwathy; Riviere, Jim E; Monteiro-Riviere, Nancy A

    2017-05-01

    This study investigated the role of nanoparticle size and surface chemistry on biocorona composition and its effect on uptake, toxicity and cellular responses in human umbilical vein endothelial cells (HUVEC), employing 40 and 80 nm gold nanoparticles (AuNP) with branched polyethyleneimine (BPEI), lipoic acid (LA) and polyethylene glycol (PEG) coatings. Proteomic analysis identified 59 hard corona proteins among the various AuNP, revealing largely surface chemistry-dependent signature adsorbomes exhibiting human serum albumin (HSA) abundance. Size distribution analysis revealed the relative instability and aggregation inducing potential of bare and corona-bound BPEI-AuNP, over LA- and PEG-AuNP. Circular dichroism analysis showed surface chemistry-dependent conformational changes of proteins binding to AuNP. Time-dependent uptake of bare, plasma corona (PC) and HSA corona-bound AuNP (HSA-AuNP) showed significant reduction in uptake with PC formation. Cell viability studies demonstrated dose-dependent toxicity of BPEI-AuNP. Transcriptional profiling studies revealed 126 genes, from 13 biological pathways, to be differentially regulated by 40 nm bare and PC-bound BPEI-AuNP (PC-BPEI-AuNP). Furthermore, PC formation relieved the toxicity of cationic BPEI-AuNP by modulating expression of genes involved in DNA damage and repair, heat shock response, mitochondrial energy metabolism, oxidative stress and antioxidant response, and ER stress and unfolded protein response cascades, which were aberrantly expressed in bare BPEI-AuNP-treated cells. NP surface chemistry is shown to play the dominant role over size in determining the biocorona composition, which in turn modulates cell uptake, and biological responses, consequently defining the potential safety and efficacy of nanoformulations.

  9. Post-translational Modification of the NKG2D Ligand RAET1G Leads to Cell Surface Expression of a Glycosylphosphatidylinositol-linked Isoform*

    Science.gov (United States)

    Ohashi, Maki; Eagle, Robert A.; Trowsdale, John

    2010-01-01

    NKG2D is an important activating receptor on lymphocytes. In human, it interacts with two groups of ligands: the major histocompatibility complex class I chain-related A/B (MICA/B) family and the UL-16 binding protein (ULBP) family, also known as retinoic acid early transcript (RAET1). MIC proteins are membrane-anchored, but all of the ULBP/RAET1 proteins, except for RAET1E and RAET1G, are glycosylphosphatidylinositol (GPI)-anchored. To address the reason for these differences we studied the association of RAET1G with the membrane. Using epitope-tagged RAET1G protein in conjunction with antibodies to different parts of the molecule and in pulse-chase experiments, we showed that the C terminus of the protein was cleaved soon after protein synthesis. Endoglycosidase H and peptide N-glycosidase treatment and cell surface immunoprecipitation indicated that most of the protein stayed in the endoplasmic reticulum, but some of the cleaved form was modified in the Golgi and transported to the cell surface. We examined the possibility of GPI anchoring of the protein in three ways: (i) Phosphatidylinositol (PI)-specific phospholipase C released the PI-linked form of the protein. (ii) The surface expression pattern of RAET1G decreased in cells defective in GPI anchoring through mutant GPI-amidase. (iii) Site-directed mutagenesis, to disrupt residues predicted to facilitate GPI-anchoring, resulted in diminished surface expression of RAET1G. Thus, a form of RAET1G is GPI-anchored, in line with most other ULBP/RAET1 family proteins. The cytoplasmic tail and transmembrane domains appear to result from gene duplication and frameshift mutation. Together with our previous results, our data suggest that RAET1G is regulated post-translationally to produce a GPI-anchored isoform. PMID:20304922

  10. Post-translational modification of the NKG2D ligand RAET1G leads to cell surface expression of a glycosylphosphatidylinositol-linked isoform.

    Science.gov (United States)

    Ohashi, Maki; Eagle, Robert A; Trowsdale, John

    2010-05-28

    NKG2D is an important activating receptor on lymphocytes. In human, it interacts with two groups of ligands: the major histocompatibility complex class I chain-related A/B (MICA/B) family and the UL-16 binding protein (ULBP) family, also known as retinoic acid early transcript (RAET1). MIC proteins are membrane-anchored, but all of the ULBP/RAET1 proteins, except for RAET1E and RAET1G, are glycosylphosphatidylinositol (GPI)-anchored. To address the reason for these differences we studied the association of RAET1G with the membrane. Using epitope-tagged RAET1G protein in conjunction with antibodies to different parts of the molecule and in pulse-chase experiments, we showed that the C terminus of the protein was cleaved soon after protein synthesis. Endoglycosidase H and peptide N-glycosidase treatment and cell surface immunoprecipitation indicated that most of the protein stayed in the endoplasmic reticulum, but some of the cleaved form was modified in the Golgi and transported to the cell surface. We examined the possibility of GPI anchoring of the protein in three ways: (i) Phosphatidylinositol (PI)-specific phospholipase C released the PI-linked form of the protein. (ii) The surface expression pattern of RAET1G decreased in cells defective in GPI anchoring through mutant GPI-amidase. (iii) Site-directed mutagenesis, to disrupt residues predicted to facilitate GPI-anchoring, resulted in diminished surface expression of RAET1G. Thus, a form of RAET1G is GPI-anchored, in line with most other ULBP/RAET1 family proteins. The cytoplasmic tail and transmembrane domains appear to result from gene duplication and frameshift mutation. Together with our previous results, our data suggest that RAET1G is regulated post-translationally to produce a GPI-anchored isoform.

  11. In Vitro Variant Surface Antigen Expression in Plasmodium falciparum Parasites from a Semi-Immune Individual Is Not Correlated with Var Gene Transcription

    Science.gov (United States)

    Tschan, Serena; Flötenmeyer, Matthias; Koch, Iris; Berger, Jürgen; Kremsner, Peter; Frank, Matthias

    2016-01-01

    Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) is considered to be the main variant surface antigen (VSA) of Plasmodium falciparum and is mainly localized on electron-dense knobs in the membrane of the infected erythrocyte. Switches in PfEMP1 expression provide the basis for antigenic variation and are thought to be critical for parasite persistence during chronic infections. Recently, strain transcending anti-PfEMP1 immunity has been shown to develop early in life, challenging the role of PfEMP1 in antigenic variation during chronic infections. In this work we investigate how P. falciparum achieves persistence during a chronic asymptomatic infection. The infected individual (MOA) was parasitemic for 42 days and multilocus var gene genotyping showed persistence of the same parasite population throughout the infection. Parasites from the beginning of the infection were adapted to tissue culture and cloned by limiting dilution. Flow cytometry using convalescent serum detected a variable surface recognition signal on isogenic clonal parasites. Quantitative real-time PCR with a field isolate specific var gene primer set showed that the surface recognition signal was not correlated with transcription of individual var genes. Strain transcending anti-PfEMP1 immunity of the convalescent serum was demonstrated with CD36 selected and PfEMP1 knock-down NF54 clones. In contrast, knock-down of PfEMP1 did not have an effect on the antibody recognition signal in MOA clones. Trypsinisation of the membrane surface proteins abolished the surface recognition signal and immune electron microscopy revealed that antibodies from the convalescent serum bound to membrane areas without knobs and with knobs. Together the data indicate that PfEMP1 is not the main variable surface antigen during a chronic infection and suggest a role for trypsin sensitive non-PfEMP1 VSAs for parasite persistence in chronic infections. PMID:27907004

  12. Co-expression of foreign proteins tethered to HIV-1 envelope glycoprotein on the cell surface by introducing an intervening second membrane-spanning domain.

    Directory of Open Access Journals (Sweden)

    Hongyun Wang

    Full Text Available The envelope glycoprotein (Env of human immunodeficiency virus type I (HIV-1 mediates membrane fusion. To analyze the mechanism of HIV-1 Env-mediated membrane fusion, it is desirable to determine the expression level of Env on the cell surface. However, the quantification of Env by immunological staining is often hampered by the diversity of HIV-1 Env and limited availability of universal antibodies that recognize different Envs with equal efficiency. To overcome this problem, here we linked a tag protein called HaloTag at the C-terminus of HIV-1 Env. To relocate HaloTag to the cell surface, we introduced a second membrane-spanning domain (MSD between Env and HaloTag. The MSD of transmembrane protease serine 11D, a type II transmembrane protein, successfully relocated HaloTag to the cell surface. The surface level of Env can be estimated indirectly by staining HaloTag with a specific membrane-impermeable fluorescent ligand. This tagging did not compromise the fusogenicity of Env drastically. Furthermore, fusogenicity of Env was preserved even after the labeling with the ligands. We have also found that an additional foreign peptide or protein such as C34 or neutralizing single-chain variable fragment (scFv can be linked to the C-terminus of the HaloTag protein. Using these constructs, we were able to determine the required length of C34 and critical residues of neutralizing scFv for blocking membrane fusion, respectively.

  13. Poleward intrusion in the northern Galician shelf

    Science.gov (United States)

    Alvarez, I.; Ospina-Alvarez, N.; deCastro, M.; Varela, M.; Gomez-Gesteira, M.; Prego, R.

    2010-05-01

    The evolution of a warm water mass related to the Iberian Poleward Current (IPC) was characterized along the northern Galician shelf in November 2008 by means of Sea Surface Temperature and wind data. It was observed that under upwelling favorable conditions water temperature decreased along the northern coast and a temperature break appeared between Cape Vilano and Cape Ortegal showing a relaxation of the poleward intrusion. The effect of the IPC was also analyzed inside the Northern Galician Rias taking into account the hydrographical and biogeochemical properties measured on November 18. Water driven by the IPC was observed close to the mouth of the rias, around Cape Estaca de Bares, causing a nutrient salts decrease. Inside the rias a slight biological activity was found near surface resulting from fluvial contributions.

  14. Cadherin-6 type 2, K-cadherin (CDH6) is regulated by mutant p53 in the fallopian tube but is not expressed in the ovarian surface.

    Science.gov (United States)

    Karthikeyan, Subbulakshmi; Lantvit, Daniel D; Chae, Dam Hee; Burdette, Joanna E

    2016-10-25

    High-grade serous ovarian cancer (HGSOC) is the most lethal gynecological malignancy and may arise in either the fallopian tube epithelium (FTE) or ovarian surface epithelium (OSE). A mutation in p53 is reported in 96% of HGSOC, most frequently at R273 and R248. The goal of this study was to identify specific gene targets in the FTE that are altered by mutant p53, but not in the OSE. Gene analysis revealed that both R273 and R248 mutant p53 reduces CDH6 expression in the oviduct, but CDH6 was not detected in murine OSE cells. p53R273H induced SLUG and FOXM1 while p53R248W did not induce SLUG and only modestly increased FOXM1, which correlated with less migration as compared to p53R273H. An oviduct specific PAX8Cre/+/p53R270H/+ mouse model was created and confirmed that in vivo mutant p53 repressed CDH6 but was not sufficient to stabilize p53 expression alone. Overexpression of mutant p53 in the p53 null OVCAR5 cells decreased CDH6 levels indicating this was a gain-of-function. SLUG knockdown in murine oviductal cells with p53R273H restored CDH6 repression and a ChIP analysis revealed direct binding of mutant p53 on the CDH6 promoter. NSC59984, a small molecule that degrades mutant p53R273H, rescued CDH6 expression. In summary, CDH6 is expressed in the oviduct, but not the ovary, and is repressed by mutant p53. CDH6 expression with further validations may aide in establishing markers that inform upon the cell of origin of high grade serous tumors.

  15. The microRNAs in an ancient protist repress the variant-specific surface protein expression by targeting the entire coding sequence.

    Directory of Open Access Journals (Sweden)

    Ashesh A Saraiya

    2014-02-01

    Full Text Available microRNAs (miRNA have been detected in the deeply branched protist, Giardia lamblia, and shown to repress expression of the family of variant-specific surface proteins (VSPs, only one of which is expressed in Giardia trophozoite at a given time. Three next-generation sequencing libraries of Giardia Argonaute-associated small RNAs were constructed and analyzed. Analysis of the libraries identified a total of 99 new putative miRNAs with a size primarily in the 26 nt range similar to the size previously predicted by the Giardia Dicer crystal structure and identified by our own studies. Bioinformatic analysis identified multiple putative miRNA target sites in the mRNAs of all 73 VSPs. The effect of miRNA target sites within a defined 3'-region were tested on two vsp mRNAs. All the miRNAs showed partial repression of the corresponding vsp expression and were additive when the targeting sites were separately located. But the combined repression still falls short of 100%. Two other relatively short vsp mRNAs with 15 and 11 putative miRNA target sites identified throughout their ORFs were tested with their corresponding miRNAs. The results indicate that; (1 near 100% repression of vsp mRNA expression can be achieved through the combined action of multiple miRNAs on target sites located throughout the ORF; (2 the miRNA machinery could be instrumental in repressing the expression of vsp genes in Giardia; (3 this is the first time that all the miRNA target sites in the entire ORF of a mRNA have been tested and shown to be functional.

  16. [Construction of RNA-containing virus-like nanoparticles expression vector with cysteine residues on surface and fluorescent decoration].

    Science.gov (United States)

    Cheng, Yang-Jian; Liang, Ji-Xuan; Li, Qing-Ge

    2005-08-01

    Site-directed mutagenesis was performed at the codon 15 of the MS2 bacteriophage coat protein gene,which had been cloned to the virus-like particles expression vector containing non-self RNA fragment. The produced expression vector,termed pARSC, was transformed to E. coli DH5alpha. The positive clones were selected and proliferated. The harvested cells were treated with sonication and the supernatant was then subjected to linear sucrose density gradients centrifugation (15% to 60%) at 32000 r/min for 4 h at 4 degrees C. The virus-like particles, VLP-Cy, were collected at 35% sucrose density. The particles were examined by transmission electron microscopy and the spherical viral particles of approximately 27 nm in diameter were found. The thiolated VLP-Cy was then chemically modified with fluorescein -5'-maleimide. The covalent fluorescent labeling was confirmed by absorption analysis, SDS-PAGE and MALDI-TOF mass spectroscopy. This is the first report of preparation of RNA-containing natural fluorescent nanoparticles. The study highlight the versatility of MS2 bacteriophage capsids as building blocks for functional nanomaterials construction for a variety of application purposes.

  17. Whole-Genome Expression Analysis of Human Mesenchymal Stromal Cells Exposed to Ultrasmooth Tantalum vs. Titanium Oxide Surfaces

    DEFF Research Database (Denmark)

    Stiehler, C.; Bunger, C.; Overall, R. W.

    2013-01-01

    of cultivation and genes upregulated by MSCs exposed to Ta and Ti were predominantly related to the processes of differentiation and transcription, respectively. Functional annotation analysis of the 1,000 temporally most significantly regulated genes suggests earlier cellular differentiation on Ta compared...... to Ti surface. Key genes related to osteogenesis and cell adhesion were upregulated by MSCs exposed to Ta. We further identified differentially regulated candidate transcription factors, e.g., NRF2, EGR1, IRF-1, IRF-8, NF-Y, and p53 as well as relevant signaling pathways, e.g., p53 and mTOR, indicating...

  18. EXPRESS-ASSESSMENT OF TOXISITY OF WATERS FROM SURFACE WATER SUPPLY SOURCES WITH THE USE OF DANIO RERIO

    OpenAIRE

    Е. Arystarkhova

    2017-01-01

    Purpose. An increase in the informativeness and shortening of the timing required for biotesting of water from surface water sources in the city of Zhytomyr with the use of Danio rerio Hamilton-Buchanan instead of the traditional test object Poecilia reticulata Peters and observation of their ethology using the «time sampling» method. Methods. Water samples were collected from reservoirs of the Teteriv river in April 2016. They were used placed in aquaria (10 dm3), where control and exper...

  19. EXPRESS-ASSESSMENT OF TOXISITY OF WATERS FROM SURFACE WATER SUPPLY SOURCES WITH THE USE OF DANIO RERIO

    Directory of Open Access Journals (Sweden)

    Е. Arystarkhova

    2017-09-01

    Full Text Available Purpose. An increase in the informativeness and shortening of the timing required for biotesting of water from surface water sources in the city of Zhytomyr with the use of Danio rerio Hamilton-Buchanan instead of the traditional test object Poecilia reticulata Peters and observation of their ethology using the «time sampling» method. Methods. Water samples were collected from reservoirs of the Teteriv river in April 2016. They were used placed in aquaria (10 dm3, where control and experimental groups were formed according to the following scheme: control group – samples with settled (24 hours tap water; experimental group D-1 - water samples from the Denishivsky reservoir; experimental group D-2 – from water intake Vidsichne. Test specimens were females of P. reticulata and D. rerio, in which the parameters of typical and non-typical motor activity and feeding behavior were determined. Fish were exposed to water for 12 hours. Water toxicity indexes were determined for both experimental and control groups. Findings. The studies showed the possibility of using the "time sampling" method for rapid assessment of the water toxicity of surface water sources of Zhytomyr with the use of P. reticulata and D. rerio, which consists in the instantaneous fixation of the number of individuals preferring different behavior acts. D. rerio were more susceptible to the combined action of pollutants in experimental waters than the traditional test object guppies regarding the test reactions of increasing the vector movement and reducing the activity of consumed feed (in 12 hours after the start of biotesting on average by 8,43% in the group D1 and by 11,71% in the group D2. The calculation of toxicity indexes based on the above-mentioned ethological acts, which exceeded 50%, detected an average level of pollution of the experimental water. Originality. For the first time, the rapid assessment of the water toxicity of surface water sources in the city

  20. Early developmental expression of a normally tumor-associated and drug-inhibited cell surface-located NADH oxidase (ENOX2) in non-cancer cells.

    Science.gov (United States)

    Cho, NaMi; Morré, D James

    2009-04-01

    Full length mRNA to a drug-inhibited cell surface NADH oxidase, tNOX or ENOX2, is present in both non-cancer and cancer cells but is translated only in cancer cells as alternatively spliced variants. ENOX2 is a growth-related protein of the external plasma membrane surface that is shed into the circulation and is inhibited by a series of quinone site inhibitors with anticancer activity. To test the possibility that ENOX2 expression might be important to early stages of non-cancer cell development, the expression of the protein was monitored in chicken embryos during their development. Polyclonal antisera to a 34 kDa human serum form of ENOX2 cross-immunoreactive with the drug-responsive NADH oxidase of chicken hepatoma cells was used. The protein was identified based on drug-responsive enzymatic activities and analyses by western blots. The drug-responsive activity was associated with plasma membranes and sera of early chicken embryos and with chicken hepatoma plasma membranes but was absent from plasma membranes prepared from livers or from sera of normal adult chickens and from late embryo stages. The findings suggest that ENOX2 may fulfill some functions essential to the growth of early embryos which are lost in late embryo stages and absent from normal adult cells but which then reappear in cancer.

  1. Retention in the Golgi apparatus and expression on the cell surface of Cfr/Esl-1/Glg-1/MG-160 are regulated by two distinct mechanisms.

    Science.gov (United States)

    Miyaoka, Yuichiro; Kato, Hidenori; Ebato, Kazuki; Saito, Shigeru; Miyata, Naoko; Imamura, Toru; Miyajima, Atsushi

    2011-11-15

    Cfr (cysteine-rich fibroblast growth factor receptor) is an Fgf (fibroblast growth factor)-binding protein without a tyrosine kinase. We have shown previously that Cfr is involved in Fgf18 signalling via Fgf receptor 3c. However, as Cfr is also known as Glg (Golgi apparatus protein)-1 or MG-160 and occurs in the Golgi apparatus, it remains unknown how the distribution of Cfr is regulated. In the present study, we performed a mutagenic analysis of Cfr to show that two distinct regions contribute to its distribution and stability. First, the C-terminal region retains Cfr in the Golgi apparatus. Secondly, the Cfr repeats in the extracellular juxtamembrane region destabilizes Cfr passed through the Golgi apparatus. This destabilization does not depend on the cleavage and secretion of the extracellular domain of Cfr. Furthermore, we found that Cfr with a GPI (glycosylphosphatidylinositol) anchor was predominantly expressed on the cell surface in Ba/F3 cells and affected Fgf18 signalling in a similar manner to the full-length Cfr, indicating that the interaction of Cfr with Fgfs on the cell surface is important for its function in Fgf signalling. These results suggest that the expression of Cfr in the Golgi apparatus and on the plasma membrane is finely tuned through two distinct mechanisms for exhibiting different functions.

  2. Cell-Surface Displayed Expression of Trehalose Synthase from Pseudomonas putida ATCC 47054 in Pichia Pastoris Using Pir1p as an Anchor Protein

    Directory of Open Access Journals (Sweden)

    Shaojie Yang

    2017-12-01

    Full Text Available Yeast cell-surface display technologies have been widely applied in the fields of food, medicine, and feed enzyme production, including lipase, α-amylase, and endoglucanase. In this study, a treS gene was fused with the yeast cell-surface anchor protein gene Pir1p by overlap PCR, the Pir1p-treS fusion gene was ligated into pPICZαA and pGAPZαA and transformed into P. pastoris GS115 to obtain recombinant yeast strains that displays trehalose synthase(TreS on its cell surface as an efficient and recyclable whole-cell biocatalyst. Firstly, the enhanced green fluorescence protein gene (egfp was used as the reporter protein to fusion the Pir1p gene and treS gene to construct the recombinant plasmids containing treS-egfg-Pir1p fusion gene, and electrotransformed into P. pastoris GS115 to analyze the surface display characteristics of fusion gene by Western blot, fluorescence microscopy and flow cytometry. The analysis shown that the treS-egfg-Pir1p fusion protein can be successfully displayed on the surface of yeast cell, and the expression level increased with the extension of fermentation time. These results implied that the Pir1p-treS fusion gene can be well displayed on the cell surface. Secondly, in order to obtain surface active cells with high enzyme activity, the enzymatic properties of TreS displayed on the cell surface was analyzed, and the fermentation process of recombinant P. patoris GS115 containing pPICZαA-Pir1p-treS and pGAPZαA-Pir1p-treS was studied respectively. The cell surface display TreS was stable over a broad range of temperatures (10–45°C and pH (6.0–8.5. The activity of TreS displayed on cell surface respectively reached 1,108 Ug−1 under PAOX1 control for 150 h, and 1,109 Ug−1 under PGAP control for 75h in a 5 L fermenter, respectively. Lastly, the cell-surface displayed TreS was used to product trehalose using high maltose syrup as substrate at pH 8.0 and 15°C. The surface display TreS cells can be recycled for

  3. Mesenchymal stromal cells induce epithelial-to-mesenchymal transition in human colorectal cancer cells through the expression of surface-bound TGF-β.

    Science.gov (United States)

    Mele, Valentina; Muraro, Manuele G; Calabrese, Diego; Pfaff, Dennis; Amatruda, Nunzia; Amicarella, Francesca; Kvinlaug, Brynn; Bocelli-Tyndall, Chiara; Martin, Ivan; Resink, Therese J; Heberer, Michael; Oertli, Daniel; Terracciano, Luigi; Spagnoli, Giulio C; Iezzi, Giandomenica

    2014-06-01

    Mesenchymal stem/stromal cells (MSC) are multipotent precursors endowed with the ability to home to primary and metastatic tumor sites, where they can integrate into the tumor-associated stroma. However, molecular mechanisms and outcome of their interaction with cancer cells have not been fully clarified. In this study, we investigated the effects mediated by bone marrow-derived MSC on human colorectal cancer (CRC) cells in vitro and in vivo. We found that MSC triggered epithelial-to-mesenchymal transition (EMT) in tumor cells in vitro, as indicated by upregulation of EMT-related genes, downregulation of E-cadherin and acquisition of mesenchymal morphology. These effects required cell-to-cell contact and were mediated by surface-bound TGF-β newly expressed on MSC upon coculture with tumor cells. In vivo tumor masses formed by MSC-conditioned CRC cells were larger and characterized by higher vessel density, decreased E-cadherin expression and increased expression of mesenchymal markers. Furthermore, MSC-conditioned tumor cells displayed increased invasiveness in vitro and enhanced capacity to invade peripheral tissues in vivo. Thus, by promoting EMT-related phenomena, MSC appear to favor the acquisition of an aggressive phenotype by CRC cells. © 2013 The Authors. Published by Wiley Periodicals, Inc. on behalf of UICC.

  4. Cell surface expression level variation between two common Human Leukocyte Antigen alleles, HLA-A2 and HLA-B8, is dependent on the structure of the C terminal part of the alpha 2 and the alpha 3 domains

    DEFF Research Database (Denmark)

    Dellgren, Christoffer; Nehlin, Jan O; Barington, Torben

    2015-01-01

    Constitutive cell surface expression of Human Leukocyte Antigen (HLA) class I antigens vary extremely from tissue to tissue and individual antigens may differ widely in expression levels. Down-regulation of class I expression is a known immune evasive mechanism used by cancer cells and viruses....... Moreover, recent observations suggest that even minor differences in expression levels may influence the course of viral infections and the frequency of complications to stem cell transplantation. We have shown that some human multipotent stem cells have high expression of HLA-A while HLA-B is only weakly...... expressed, and demonstrate here that this is also the case for the human embryonic kidney cell line HEK293T. Using quantitative flow cytometry and quantitative polymerase chain reaction we found expression levels of endogenous HLA-A3 (median 71,204 molecules per cell) 9.2-fold higher than the expression of...

  5. Protein kinase A stimulates Kv7.1 surface expression by regulating Nedd4-2-dependent endocytic trafficking

    DEFF Research Database (Denmark)

    Andersen, Martin Nybo; Hefting, Louise Leth; Steffensen, Annette Buur

    2015-01-01

    The potassium channel Kv7.1 plays critical physiological roles in both heart and epithelial tissues. In heart, Kv7.1 and the accessory subunit KCNE1 forms the IKs current, which is enhanced by PKA mediated phosphorylation. The observed current increase requires both phosphorylation of Kv7.......1 and the presence of KCNE1. However, PKA also stimulates Kv7.1 currents in epithelial tissues, such as colon, where the channel does not co-assemble with KCNE1. Here, we demonstrate that PKA activity significantly impacts the subcellular localization of Kv7.1 in Madin Darby Canine Kidney cells. While PKA inhibition...... reduced the fraction of channels at the cell surface, PKA activation increased it. We show that PKA inhibition lead to intracellular accumulation of Kv7.1 in late endosomes/lysosomes. By mass spectroscopy we identified eight phosphorylated residues on Kv7.1, however, none appeared to play a role...

  6. Quantification of cell surface receptor expression in live tissue culture media using a dual-tracer stain and rinse approach

    Science.gov (United States)

    Xu, Xiaochun; Sinha, Lagnojita; Singh, Aparna; Yang, Cynthia; Xiang, Jialing; Tichauer, Kenneth M.

    2015-03-01

    Immunofluorescence staining is a robust way to visualize the distribution of targeted biomolecules invasively in in fixed tissues and tissue culture. Despite the fact that these methods has been a well-established method in fixed tissue imaging for over 70 years, quantification of receptor concentration still simply assumes that the signal from the targeted fluorescent marker after incubation and sufficient rinsing is directly proportional to the concentration of targeted biomolecules, thus neglecting the experimental inconsistencies in incubation and rinsing procedures and assuming no, nonspecific binding of the fluorescent markers. This work presents the first imaging approach capable of quantifying the concentration of cell surface receptor on cancer cells grown in vitro based on compartment modeling in a nondestructive way. The approach utilizes a dual-tracer protocol where any non-specific retention or variability in incubation and rinsing of a receptor-targeted imaging agent is corrected by simultaneously imaging the retention of a chemically similar, "untargeted" imaging agent. Various different compartment models were used to analyze the data in order to find the optimal procedure for extracting estimates of epidermal growth factor receptor (EGFR) concentration (a receptor overexpressed in many cancers and a key target for emerging molecular therapies) in tissue cultures with varying concentrations of human glioma cells (U251). Preliminary results demonstrated a need to model nonspecific binding of both the targeted and untargeted imaging agents used. The approach could be used to carry out the first repeated measures of cell surface receptor dynamics during 3D tumor mass development, in addition to the receptor response to therapies.

  7. Human platelet glycoprotein Ia. One component is only expressed on the surface of activated platelets and may be a granule constituent

    International Nuclear Information System (INIS)

    Bienz, D.; Clemetson, K.J.

    1989-01-01

    Glycoprotein Ia (GP Ia) is a relatively minor component of human blood platelets thought to be a receptor involved in collagen-induced platelet activation. However, some difficulties exist with the definition of this glycoprotein. The expression of GP Ia on resting (prostacyclin analogue-treated) and thrombin-activated platelets was compared by surface labeling with 125 I-lactoperoxidase. Intact platelets or platelets solubilized in sodium dodecyl sulfate were labeled with periodate/[ 3 H]NaBH 4 . Analysis on two-dimensional isoelectric focusing/sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels showed that GP Ia is very poorly labeled in resting platelets. After activation a new spot (GP Ia*) appears with the same relative molecular mass as GP Ia under reducing conditions. GP Ia and Ia* can be clearly separated by two-dimensional nonreduced/reduced gel electrophoresis. Therefore, two glycoproteins which have been termed GP Ia exist in platelets with similar molecular weight and pI under reducing conditions. One of these (GP Ia*) is only surface-labeled when platelets are activated, indicating that it is only exposed on the surface of activated platelets. Supernatant from activated platelets contains this glycoprotein as well as other granule components. This glycoprotein is missing in platelets from two patients with collagen-response defects

  8. Nasal immunization of mice with Lactobacillus casei expressing the Pneumococcal Surface Protein A: induction of antibodies, complement deposition and partial protectio