WorldWideScience

Sample records for characterizing metallogenic factors

  1. Hyperspectral information identification for characterizing metallogenic factors of Taoshan uranium ore-field

    International Nuclear Information System (INIS)

    Zhang Jielin

    2007-01-01

    The application of high resolution remote sensing technology to uranium mineralization is discussed in detail. By utilizing field hyperspectrum measurement method (VIS/NIR-SWIR-FIR) and fusion technology of ETM image and Radarsat data, the spectral characteristic of uranium mineralization factors including rock mass, ore-control fault and hydrothermal alteration are studied in Taoshan granite type uranium deposits. The main ore-control structure has been identified based on remote sensing image feature. Those results and experiences provide the new remote sensing approaches to the prospecting and evaluation of uranium resources. (authors)

  2. Regional metallogenic essential factor of granite-type uranium deposits in Guangdong province

    International Nuclear Information System (INIS)

    Pan Yongzheng

    1987-12-01

    The uranium origin, activation region, red basin, and fault depressed zone constitute the regional metallogenic essential factor of the four united like one granite-type uranium deposits in the post-Caledonian rise area in China. In the development of sub-geosyncline in the Caledonian, the clastic formation with widely deposited carbon, silicon, mud rich bearing organic matter, which drow a great amount of uranium formed the uranium-bearing system in the Sinian-Cambrian period. The magmagranite activation in a large scale in the Indosinian-Yenshanian period caused the continental crust to be suffered strong reformation and the uranium-bearing basement system to be eroded and remelted, and formed the rich uranium granite body. The multiple structure-magmatic movement further made the uranium in the rock body suffered the endogenic, structure, supergene active reformation, and produced mobile uranium concentrated area. Under the dry and hot paleoclimate condition in the Cretaceous-Tertiary period, strong weathering and hot water leaching forced uranium to be concentrated into the 'rock origin activation' type uranium deposits in the fault depressed zone

  3. Uranium metallogenic features and prospecting potentialities in the areas around Shabazi uranium deposit in Nanling metallogenic belt

    International Nuclear Information System (INIS)

    Yang Shanghai

    2008-01-01

    Based on the actuality of exploration and research on Shabazi uranium deposit in Nanling metallogenic belt, the author analyzes and summarizes uranium metallogenic features of the deposit. Under the direction of modern metallogenic theories of uranium deposit, such as deep-source mineralization and deep prospecting for uranium deposits, it is shown that there is great mineralization and prospecting potentiality in the areas around Shabazi uranium deposit and high attention importance should be paid to the areas in the future exploration according to the synthetical analysis on geologic background of the deposit, uranium mineralization features, ore-controlling factors and systematic data of geology. (authors)

  4. Granite metallogenic specialization study based on RS information model-A case of hydrothermal uranium and tungsten deposits in Nanling region

    International Nuclear Information System (INIS)

    Huang Hongye; Qin Qiming

    2009-01-01

    According to the granite hydrothermal metallogenic principle, metallogenic specialization information model for uranium producing and tungsten producing granites in Nanling region is built up and the group factor system of granite metallogenic specialization is initially proposed by using RS information model. On the basis of the above aspects, the geographical index and coefficients of information model of granite metallogenic specialization are respectively analyzed, metallogenic specialization discrimination criterion is built up. After the non-discriminatory massif is forecasted, the results are basically accordant with geological fact, at the same time they are used in the geological metallogenic research, which indicates that metallogenic specialization information model is objective and operative, realizes quantitative appraisal on metallogenic specialization and provides a scientific basis for further discriminating the ore-forming massif. (authors)

  5. Overview of Qatecalovo-Kulaming uranium metallogenic region

    International Nuclear Information System (INIS)

    Zhao Fengmin

    2011-01-01

    Qatecalovo-Kulaming uranium metallogenic region is the earliest discovered volcanic-type uranium metallogenic region in the world, which which is located in the Kulaming uplift of Middle Tianshan Variscan orogenic belt. Ten deposits have been discovered in the mine area, over 20000 t metallic uranium has been mined out. Uranium deposits mainly occurred in Late Palaeozoic volcanic depression, and the volcanic rocks are characterized by the great lithologic change from andesite to rhyolite. Mineralization age is from 260 to 270 Ma. The tectonic unit in which the metallogenic region is located can extend eastward into China. Therefore, the ore-forming condition study on this tectonic unit can be used to guide the prospection for Palaeozoic volcanics uranium deposit in northwestern China. (author)

  6. Metallogenic events in the Coastal Cordillera of Copiapo region, northern Chile (26-28o S)

    International Nuclear Information System (INIS)

    Diaz, A.; Vivallo, W

    2001-01-01

    The metallogenic province of the Coastal Cordillera in the Copiapo region is characterized by Cu, Cu-Au,Cu-Fe-Au, Fe, Ag and Au deposits. The ore deposits are ranging in ages between Middle to Late Jurassic (1) and Early Cretaceous (2). Results of a geochronology program for ore deposits in this metallogenic province, in addition to existing geochronological control of the ore deposits host rocks (mainly intrusive) allowed to define four different Metallogenic Epochs which could be extended to other zones in the Coastal Cordillera (au)

  7. Preliminary discussion on uranium metallogenic models of China's in-situ leachable sandstone-type uranium deposits

    International Nuclear Information System (INIS)

    Zhang Jindai; Xu Gaozhong; Chen Anping; Wang Cheng

    2005-01-01

    By comprehensively analyzing metallogenic environments and main ore-controlling factors of important uranium metallogenic regions of in-situ leachable sandstone-type uranium deposits at the southern margin of Yili basin, at the south-western margin of Turpan-Hami basin and in the northeastern Ordos basin, the authors of this paper discuss the metallogenic models of China's in-situ leachable sandstone-type uranium deposits, and suggest that the interlayer oxidation zone type uranium deposits in Yili and Turpan-Hami basins are basically controlled by favourable structures, sedimentary formations and interlayer oxidation zone, and are characterized by multistage uranium concentration, namely the uranium pre-concentration of ore-hosting sedimentary formation, the uranium ore-formation in the stage of supergenic epigenetic reworking, and the further superimposition enrichment of post-ore tectonic activity. However, the interlayer oxidation zone type uranium deposit in the northeastern Ordos was formed after the formation of the secondary reduction. So, paleo-interlayer oxidation zone type uranium mineralization has the mineralization size much greater than the former two. (authors)

  8. Brief analysis on uranium metallogenic potential in Wunikeng area

    International Nuclear Information System (INIS)

    Wang Binghua

    2010-01-01

    Through the geological mapping, gamma total equivalent weight survey in Wunikeng area, the tectonics and gamma anomalies discovered in this area are analyzed, uranium metallogenic potential is analyzed by combining the regional metallogenic condition, contrasting the metallogenic geological background, massif condition, tectonic framework from these aspects of the regional geological background, massif condition on the work area, tectonic framework, etc.. (authors)

  9. Tectonic and metallogenic model for northeast Asia

    Science.gov (United States)

    Parfenov, Leonid M.; Nokleberg, Warren J.; Berzin, Nikolai A.; Badarch, Gombosuren; Dril, Sergy I.; Gerel, Ochir; Goryachev, Nikolai A.; Khanchuk, Alexander I.; Kuz'min, Mikhail I.; Prokopiev, Andrei V.; Ratkin, Vladimir V.; Rodionov, Sergey M.; Scotese, Christopher R.; Shpikerman, Vladimir I.; Timofeev, Vladimir F.; Tomurtogoo, Onongin; Yan, Hongquan; Nokleberg, Warren J.

    2011-01-01

    This document describes the digital files in this report that contains a tectonic and metallogenic model for Northeast Asia. The report also contains background materials. This tectonic and metallogenic model and other materials on this report are derived from (1) an extensive USGS Professional Paper, 1765, on the metallogenesis and tectonics of Northeast Asia that is available on the Internet at http://pubs.usgs.gov/pp/1765/; and (2) the Russian Far East parts of an extensive USGS Professional Paper, 1697, on the metallogenesis and tectonics of the Russian Far East, Alaska, and the Canadian Cordillera that is available on the Internet at http://pubs.usgs.gov/pp/pp1697/. The major purpose of the tectonic and metallogenic model is to provide, in movie format, a colorful summary of the complex geology, tectonics, and metallogenesis of the region. To accomplish this goal four steps were taken: (1) 13 time-stage diagrams, from the late Neoproterozoic (850 Ma) through the present (0 Ma), were adapted, generalized, and transformed into color static time-stage diagrams; (2) the 13 time-stage diagrams were placed in a computer morphing program to produce the model; (3) the model was examined and each diagram was successively adapted to preceding and subsequent diagrams to match the size and surface expression of major geologic units; and (4) the final version of the model was produced in successive iterations of steps 2 and 3. The tectonic and metallogenic model and associated materials in this report are derived from a project on the major mineral deposits, metallogenesis, and tectonics of the Northeast Asia and from a preceding project on the metallogenesis and tectonics of the Russian Far East, Alaska, and the Canadian Cordillera. Both projects provide critical information on bedrock geology and geophysics, tectonics, major metalliferous mineral resources, metallogenic patterns, and crustal origin and evolution of mineralizing systems for this region. The major

  10. Metallogenic epoch of the Jiapigou gold belt, Jilin Province, China

    Indian Academy of Sciences (India)

    Metallogenic epoch of the Jiapigou gold belt, Jilin Province, China: ... The Jiapigou gold belt is located on the northern margin of the North China Craton, and is one of the ... 29, Xueyuan Road, Beijing 100083, People's Republic of China.

  11. Metallogenic aspects of Itu intrusive suite

    International Nuclear Information System (INIS)

    Amaral, G.; Pascholati, E.M.

    1990-01-01

    The integrated use of geological, geochemical, geophysical and remote sensing data is providing interesting new information on the metallogenic characteristics of the Itu Intrusive Suite. During World War II, up to 1959, a wolframite deposit was mined near the border of the northernmost body (Itupeva Granite). This deposit is formed by greisen veins associated with cassiterite and topaz, clearly linked with later phases of magmatic differentiation. Generally those veins are related to hydrothermal alteration of the granites and the above mentioned shear zone. U, Th and K determinations by field and laboratory gammaspectrometry were used for regional distribution analysis of those elements and its ratios and calculation of radioactivity heat production. In this aspects, the Itupeva Granite is the hottest and presents several anomalies in the Th/U ratio, indicative of late or post magmatic oxidation processes. (author)

  12. Some tentative idea about uranium metallogenic prognosis on the basis of application of GIS

    International Nuclear Information System (INIS)

    Li Jianwei; Li Xianfu

    1998-01-01

    Geographic information system (GIS) is a new borderline subject. The principal components of MAPGIS are introduced, and the data needed, main procedures and outstanding advantages in uranium metallogenic prognosis applying MAPGIS are discussed. For uranium metallogenic areas with detailed research basis and abundant data, it is of great prospects to apply MAPGIS for metallogenic prognosis

  13. Discussion of metallogenic substance source of Xiangshan uranium orefield

    International Nuclear Information System (INIS)

    Shao Fei; Tang Xiangsheng; Zou Maoqin; Hu Maomei; He Xiaomei; Chen Xiaoming; Xu Hengli

    2008-01-01

    Analysis of uranium source is a key problem for study on uranium deposit genesis. Based on analysis of general implication for determination of uranium source on distribution characteristics of regional uranium abundance, according to temporal and spatial evolution of regional metallogenic substances in process of geological history, and combining with indication for analysis of uranium source by Pb isotopic composition of ores and REE geochemistry of both rocks and ores in Xiangshan orefield, Lower Cambrian strata are determined as regional uranium source bed, Xiangshan volcanic basin is the accumulation area for regional metallogenic substances, magma and hydrothermal solution of post magmatism are media for uranium. Magmatism realizes uranium migration from 'source' to 'accumulation'. In process of magmatic evolution, uranium transformed into gas phase to provide substance base for uranium mineralization. Fluid-rock interaction of post magmatism also promoted some uranium from schist of the basement and rhyodacite into metallogenic solution. (authors)

  14. Metallogenic hydrothermal solution system of post volcanic magma in Xiangshan ore field

    International Nuclear Information System (INIS)

    Xu Hengli; Shao Fei; Zou Maoqin

    2009-01-01

    This paper has systematically described uranium metallogenic characteristics of Xiangshan ore field.Sources of metallogenic materials are discussed in different temporal and spatial scale. Combining with background analysis of metallogenic tectonic-magmatic-geodynamics, formation and evolution of metallogenic hydrothermal solution system in Xiangshan volcanic basin are studied. Metallogenic hydrothermal solution system in Xiangshan ore field is considered as the objective product of systematic evolution of hydrothermal solution in post volcanic magma constrained by regional tectonic environment. In time scale, metallogenic hydrothermal solution system developed for about 50 Ma, but its active spaces varied in different time domains. So temporal and spatial distribution of uranium mineralization is constrained. Further exploration for the ore field is also suggested in this paper. (authors)

  15. Analysis on metallogenic conditions of paleochannel sandstone type uranium deposits in kelulun region

    Energy Technology Data Exchange (ETDEWEB)

    Mingyue, Feng [Beijing Research Inst. of Uranium Geology, Beijing (China)

    1999-07-01

    On the basis of comprehensively analyzing metallogenic conditions of paleochannel sandstone type uranium deposits, the author discusses regional geologic background, characteristics of the basement and sedimentary cover of Kelulun basin and Huchawula-Hulun Lake basin, and the metallogenic potential of paleochannel sandstone type uranium deposits is proposed.

  16. Analysis on metallogenic conditions of paleochannel sandstone type uranium deposits in kelulun region

    International Nuclear Information System (INIS)

    Feng Mingyue

    1999-01-01

    On the basis of comprehensively analyzing metallogenic conditions of paleochannel sandstone type uranium deposits, the author discusses regional geologic background, characteristics of the basement and sedimentary cover of Kelulun basin and Huchawula-Hulun Lake basin, and the metallogenic potential of paleochannel sandstone type uranium deposits is proposed

  17. A comment on the metallogenic theory of exogenetic uranium ore deposits

    International Nuclear Information System (INIS)

    Liu Xiaodong; Yu Dagan

    2010-01-01

    The theory of exogenetic sandstone-type uranium followed the form process of construction in the early time, and discussed the uranium metallization by chemical enrichment during the phase of syn-deposition and diagenesis. Later, the epigenetic theory was put forward by emphasizing hydrodynamic influence on mineralization. The idea of uranium mineralization in open systems is a renovated metallogenic theory for uranium, which confirms the role of exogenesis playing in uranium mineralization. For open systems, this paper underlines that, as the most critical factors for uranium mineralization, both uranium sources and reduce agents should be open to form a dual-open system. Uranium ore deposits in the tectonic zone of eastern China formed in dual-open system, where uranium has been associated with coal, petroleum and natural gas in the sandstone sequence. (authors)

  18. Metallogenic characteristics of volcanic hydrothermal type U-Au-polymetallic deposits in Yanshan-Liaoning region

    International Nuclear Information System (INIS)

    Luo Yi; Zhou Dean; He Yiqiang; Tao Quan; Xia Yuliang; Cui Huanmin; Zhu Deling

    1996-03-01

    Yanshan-Liaoning area is located in the east part of the northern margin of North-China platform. It is a famous metallogenic region of Mesozoic volcanic hydrothermal type U-Au-polymetallic deposits in the country. The metallogenesis is controlled by a united Late Mesozoic continental taphrogenic volcano-magmatic activity. The metallogenic epochs are concentrated in Late Jurassic-Early Cretaceous periods. The metallogenic media are moderate and moderate-low temperature volcanic hydrothermal solutions originated from the mixing of volcano-magmatic water, metamorphic water and atmospheric water. The ore-forming materials are mainly derived from enrichment type upper mantle and lower crust. (8 refs., 5 figs.)

  19. Analysis on uranium mineralization-formation condition and prospecting potential in Xidamingshan metallogenic belt

    International Nuclear Information System (INIS)

    Li Zhixing; Qi Fucheng; He Zhongbo; Zhang Zilong

    2012-01-01

    There are many different opinions about the source of uranium and metallogenic mechanism in Xidamingshan metallogenic belt. therefore it blocked uranium resources potential evaluation and ore exploration. Through absorb in- formation and investigate Daxin deposit and systematic analysis on samples. It is believed that uranium source mainly came from Cambrian System, a little came from the deep. The Devonian System is favorable room for saving ores in addition to be pre-concentrated room for uranium. Also, there are new cognition about uranium metallogenic mechanism, establish ore-forming series of Xidamingshan metallogenic belt, It is proposed that uranium mineralization have experienced 4 stages; It is cleared that hydrothermal fluid superposition transform type uranium deposit is main genetic type, ancient karst accumulate type is secondary genetic type, the later is formed by leaching the former and then precipitate, enrichment in ancient karst congeries, which is formed nearby faults and with the movement of Xishan structural movement. It is proven that metallogenic potential of Xidamingshan metallogenic belt is good. Tectonic rock controlled by subsidiary fracture nearby regional main fault, which connected with the Cambrian System and the Devonian System, and the deep of the deposit is guideline of mineral prospecting next stage. (authors)

  20. Paired Magmatic-Metallogenic Belts in Myanmar - an Andean Analogue?

    Science.gov (United States)

    Gardiner, Nicholas; Robb, Laurence; Searle, Michael; Morley, Christopher

    2015-04-01

    contrasting minerals endowment. The Mogok-Mandalay-Mergui (MMM) Belt hosts crustal-melt S-type granites with significant tin-tungsten mineralization, and contains the historically major tungsten deposit of Mawchi. The Wuntho-Popa Arc comprises I-type granites and granodiorites with porphyry-type copper-gold and epithermal gold mineralization, and includes the world-class Monywa copper mine. Recent U-Pb radiometric age dating has shown the potential for the two belts to be both active from the Late Cretaceous to Eocene. The spatial juxtaposition of these two sub-parallel belts, the implication of contemporary magmatism, and their distinct but consistent metallogenic endowment bears strong similarities to the metallogenic belts of the South American Cordillera. Here we investigate whether they together represent the magmatic and metallogenic expression of an Andean-type setting in Myanmar during the subduction of Neo-Tethys. In this analogue the Wuntho-Popa Arc represents a proximal I-type magmatic belt sited immediately above the eastwards-verging Neo-Tethys subduction zone. Exhibiting porphyry-type copper-gold and epithermal gold mineralization, this would therefore be the Myanmar equivalent of the Andean coastal copper belts. Conversely, the parallel MMM Belt, comprised of more distal crustal-melt S-type tin granites, would have an analogue in the Bolivian tin belt.

  1. Metallogenic relationships to tectonic evolution - the Lachlan Orogen, Australia

    Science.gov (United States)

    Bierlein, Frank P.; Gray, David R.; Foster, David A.

    2002-08-01

    Placing ore formation within the overall tectonic framework of an evolving orogenic system provides important constraints for the development of plate tectonic models. Distinct metallogenic associations across the Palaeozoic Lachlan Orogen in SE Australia are interpreted to be the manifestation of interactions between several microplates and three accretionary complexes in an oceanic back-arc setting. In the Ordovician, significant orogenic gold deposits formed within a developing accretionary wedge along the Pacific margin of Gondwana. At the same time, major porphyry Cu-Au systems formed in an oceanic island arc outboard of an evolved magmatic arc that, in turn, gave rise to granite-related Sn-W deposits in the Early Silurian. During the ongoing evolution of the orogen in the Late Silurian to Early Devonian, sediment-hosted Cu-Au and Pb-Zn deposits formed in short-lived intra-arc basins, whereas a developing fore-arc system provided the conditions for the formation of several volcanogenic massive sulphide deposits. Inversion of these basins and accretion to the Australian continental margin triggered another pulse of orogenic gold mineralisation during the final consolidation of the orogenic belt in the Middle to Late Devonian.

  2. Metallogenesis and metallogenic model of Nuheting uranium deposit in Erlian Basin

    International Nuclear Information System (INIS)

    Li Hongjun; Kuang Wenzhan

    2010-01-01

    Based on the study on geological characteristics, metallogesis and geochemical features in Nuheting uranium deposit, it is considered that the deposit belongs to syn-sedimentary and epigenetic reworking type. The deposit position was controlled by the lake area developed during Erlian period in Late Cretaceous. The metallognesis has experienced three stages, they are syn-sedimentary metallogenesis, epigenetic reworking metallogenesis and exogenic metallogenesis. The ore-forming ages are respectively 85 Ma, (41±5)Ma and 6-13 Ma. Based on the summary of metallogenic geological features,metallogenesis and geochemical features, the metallogenic model of Nuheting uranium deposit has been established. (authors)

  3. Discussion on metallogenic prospect of sandstone-type uranium deposit in Yabulai basin

    International Nuclear Information System (INIS)

    Wang Lianshe; Li Xiangping

    2003-01-01

    Based on characteristics of initial basin type and tectonic reworking process, this article analyses the distribution features of depositional system and subsequent alteration of the target horizon of sandstone-type uranium deposits in Yabulai basin. Guided by prognostic criteria of sandstone-type uranium deposits, authors suggest that the post-depositional tectonic reworking in the basin was quite intense, and uranium metallogenic prospects are unfavorable. However, the Lower Cretaceous in the paleo-slope at the middle of the basin does show certain metallogenic prospects for sandstone-type uranium deposits

  4. Main geologic characteristics and metallogenic models of uranium deposits in Zhejiang

    International Nuclear Information System (INIS)

    Tang Qitao

    2000-01-01

    Uranium resources in Zhejiang is abundant with numerous mineralization types. According to the genesis they can be classified into: sedimentary-reworking type, hydrothermal type and infiltration type. The author briefly describes main geologic characteristics and metallogenic models of different type uranium deposits

  5. Study on integrated evaluation of sandstone-hosted uranium metallogenic potential in southern Yili basin

    International Nuclear Information System (INIS)

    Han Shaoyang; Ke Dan; Xu Jianguo; Zheng Enjiu; Li Shengxiang

    2008-01-01

    Plenty of geological data have been accumulated during mineral resource survey in China; under the guidance of new metallogenic theories, it is an important task of how to use these data most effectively for the new cycle uranium survey. In this paper, the flow of establishing the integrated mineral deposits prospecting model for sandstone-hosted uranium deposits is put forward. Based on studying geologic, hydrogeologic and regional geophysical field characteristics of representative uranium deposits No. 512 in southern Yili basin, its multi-source information descriptive model has been established, from which 512-type integrated prospecting models of sandstone-hosted uranium orefield and deposits are summarized. According to the established integrated prospecting models, the metallogenic information extraction of sandstone-hosted uranium deposits has completed in the study area. Finally, the integrated quantitative evaluation of sandstone-hosted uranium metallogenic potential is performed by using the evidence weighing method to integrate middle scale multi-source metallogenic information in the southern Yili basin, and good prediction effect is obtained. (authors)

  6. Application of Research on the Metallogenic Background in the Assessment of Mineral Resources Potentiality

    Science.gov (United States)

    Jia, D.; Feng, Y.; Liu, J.; Yao, X.; Zhang, Z.; Ye, T.

    2017-12-01

    1. Working BackgroundCurrent Status of Geological Prospecting: Detecting boundaries and bottoms, making ore search nearby; Seeing the stars, not seeing the Moon; Deep prospecting, undesirable results. The reasons of these problems are the regional metallogenic backgroud unclear and the metallogenic backgroud of the exploration regions unknown. Accordingly, Development and Research Center, CGS organized a geological setting research, in detail investigate metallogenic geological features and acquire mineralization information. 2. Technical SchemeCore research content is prediction elements of Metallogenic Structure. Adopt unified technical requirements from top to bottom, and technical route from bottom to top; Divide elements of mineral forecast and characteristics of geological structure into five elements for research and expression; Make full use of geophysical, geochemical and remote sensing inferences for the interpretation of macro information. After eight years the great project was completed. 3. Main AchievementsInnovation of basic maps compilation content of geological background, reinforce of geological structure data base of potentiality valuation. Preparation of geotectonic facies maps in different scales and professions, providing brand-new geologic background for potentiality assessment, promoting Chinese geotectonic research to the new height. Preparation of 3,375 geological structure thematic base maps of detecting working area in 6 kinds of prediction methods, providing base working maps, rock assemblage, structure of the protolith of geologic body / mineralization / ore controlling for mineral prediction of 25 ores. Enrichment and development of geotectonic facies analysis method, establishment of metallogenic background research thoughts and approach system for assessment of national mineral resources potentiality for the first time. 4. Application EffectOrientation——More and better results with less effort. Positioning——Have a definite

  7. Study on fractal characteristics of remote sensing image in the typical volcanic uranium metallogenic areas

    International Nuclear Information System (INIS)

    Pan Wei; Ni Guoqiang; Li Hanbo

    2010-01-01

    Computing Methods of fractal dimension and multifractal spectrum about the remote sensing image are briefly introduced. The fractal method is used to study the characteristics of remote sensing images in Xiangshan and Yuhuashan volcanic uranium metallogenic areas in southern China. The research results indicate that the Xiangshan basin in which lots of volcanic uranium deposits occur,is of bigger fractal dimension based on remote sensing image texture than that of the Yuhuashan basin in which two uranium ore occurrences exist, and the multifractal spectrum in the Xiangshan basin obviously leans to less singularity index than in the Yuhuashan basin. The relation of the fractal dimension and multifractal singularity of remote sensing image to uranium metallogeny are discussed. The fractal dimension and multifractal singularity index of remote sensing image may be used to predict the volcanic uranium metallogenic areas. (authors)

  8. ALPINE MAGMATIC-METALLOGENIC FORMATIONS OF THE NORTHWESTERN AND CENTRAL DINARIDES

    Directory of Open Access Journals (Sweden)

    Jakob Pamić

    1997-12-01

    Full Text Available In the paper are presented basic geological, petrologieca1, geochemi-cal and mineral deposit data for five main magmatic-metallogenic formations of the northwestern and central Dinarides: (lThe Permo Triassic rifting related andesite-diorite formations; (2 The Jurassic-Lower Cretaceous accretionary (ophiolite formations; (3 The Upper Cretaceous-Paleogene subduction related basalt-rhyohite formations; (4 The Paleogene collisional granite formations, and (5 The Oligo-cene-Neogene postsubduction andesite formations. All these magmatic-metallogenic formations originated in different geotectonic settings during the Alpine evolution of the Dinaridic parts of thc Tethys and the postorogenic evolution of the Paratethys and the Pannonian Basin, respectively.

  9. Uranium metallogenic geological conditions in the south central section of da hinggan mountains

    International Nuclear Information System (INIS)

    Wang Qing; Liu Qing

    2014-01-01

    The south central section of Da Hinggan Mountains, where the Zha Lantun prospecting zones of volcanic type uranium ore, is a high density concentrated distribution area of uranium and polymetallic mineral. This article elaborated uranium metallogenic geological conditions in the south central section of Da Hinggan Mountain, from the tectonic conditions, the source of uranium, the heat source, the space for ore-forming, hydrothermal alteration, the mineralization, and ect. This area has a good prospecting foreground and potentiality. (authors)

  10. Uranium deposits of Gabon and Oklo reactors. Metallogenic model for rich deposits of the lower proterozoic

    International Nuclear Information System (INIS)

    Gauthier-Lafaye, F.

    1986-05-01

    The geology of the Franceville basin (Gabon) is examined: stratigraphy, tectonics and geodynamics. The mobile zone of the Ogooue is specially studied: lithology, metamorphism and tectonics, isotopic geochronologic data are given. The different uranium deposits are described. A whole chapter is devoted to the study of Oklo natural nuclear reactor. A metallogenic model is proposed evidencing conditions required for deposit genesis. Tectonics, microstructures sedimentology, organic matter, diagenesis and uraniferous mineralizations are examined [fr

  11. Metallogenic aspects of the feldspars and micas geochemistry in pegmatite from Alto-Ligonha (Mocambique)

    International Nuclear Information System (INIS)

    Neves, J.M.C.

    1990-01-01

    This paper deals with metallogenic aspects concerning the huge Alto Ligonha pegmatite Province. The geological setting of the pegmatites is briefly reviewed and the metamorphic grade of the country rocks of the pegmatites, ranging from granulitic to greenschist facies, has been considered. The economically most interesting pegmatites are those emplaced within rocks with lighter metamorphism. The available geochronological data allow us to link, the most interesting pegmatites from Alto Ligonha, to the Pan-African granitoid magmatism, about 500 Ma ago. (author)

  12. Analysis on uranium resource situations and metallogenic potential of Heyuan mineralization belt of Guangdong province

    International Nuclear Information System (INIS)

    Chen Zhuhai; Zheng Mingliang; Song Shizhu; Liang Yewu; Zhao Wei

    2008-01-01

    Heyuan mineralization belt is a structure-magmatic activities belt which is charcterized by strong plastic deformation and shearing. The squeeze (overthrust) structure is formed early. Later stretch produced slide shovel-shaped normal fault which control the red basin and uranium mineralization. Comprehensive study shows that this area is of favorable uranium metallogenic condition due to the rich uranium source and higher degree of geology work, it is a target for the new round resources exploration. (authors)

  13. Sequence stratigraphy and uranium metallogenic characteristics in Xinminpu group, lower cretaceous, in Gongpoquan basin

    International Nuclear Information System (INIS)

    Gong Binli; Wang Jingping

    2006-01-01

    Characteristics of sequence stratigraphy, the distribution and the geologic time of sequence, boundary features and internal composition, the sedimentary facies, as well as the characteristics of interlayer oxidation zone and U-mineralization are expounded. It is suggested that the Gongpoquan area is of certain prospect for U-prospecting, and uranium metallogenic conditions are not favorable in the Beiluotuoquan area, and the Nalinsuhuai area has no prospect either. (authors)

  14. Metallogenic characteristics, model and exploration prospect for the paleo-interlayer-oxidation type sandstone-hosted uranium deposits in China

    International Nuclear Information System (INIS)

    Huang Jingbai; Li Shengxiang

    2007-01-01

    In this paper, the paleo-interlayer-oxidation type sandstone-hosted uranium deposits occurred in the Meso-Cenozoic continental basins in China are divided into 3 subtype, they are stratum over lapping buried subtype, structure-uplifting destroy subtype and faulted-folding conserved subtype. The metallogenic characteristics, metallogenic model and exploration prospect for these 3 subtypes uranium deposits are discussed. It is proposed that the paleo-interlayer-oxidation type sandstone-hosted uranium deposits, besides the recent interlayer oxidation type sandstone-hosted uranium deposits, are of great prospecting potential in the Meso-Cenozoic continental basins in China. Therefore, the metallogenic theory of these types uranium deposits should be conscientiously summarized and replenished continuously so as to propel forward the exploration of the sandstone-hosted uranium deposits in China. (authors)

  15. Analysis on tectonomagmatic evolution and metallogenic geological conditions of rich-large uranium deposits in Nanling uranium-polymetal metallogenic belt, South Jiangxi province

    International Nuclear Information System (INIS)

    Chen Guihua

    2001-01-01

    Nanling is a complicated tectonomagmatic belt. It is also an important uranium-polymetallic metallogenic belt. On the basis of the research achievements of previous researchers, through the detailed stratigraphic, lithologic, litho-paleo-geographic, geochronologic and geochemical studies and the processing of gravimetric and magnetic data, as well as the information extraction of remote sensing data, the authors come to the following conclusions: (1) The Sannan-Xunwu fault is a long-lived (from the Early Sinian to recent time) EW-trending fault; (2) The earth crust in Nanling experienced three time opening-closing with different scales; (3) The Anyuan-Xunwu area is a large thermo-upwelling extensional structure; (4) The above area is considered to be favourable for the formation of rich-large uranium deposit

  16. Metallogenic geologic prerequisites of sandstone-type uranium deposits and target area selection. Taking Erlian and Ordos basins as examples

    International Nuclear Information System (INIS)

    Chen Fazheng

    2002-01-01

    Sandstone-type uranium deposit is the main target of recent uranium prospecting and exploration. According to the metallogenic characteristics, sandstone-type uranium deposits are divided into three groups: paleo-channel type, interlayer oxidation zone type and phreatic interlayer oxidation type. The author makes an analysis on the geologic prerequisites of the three types of uranium deposits, the similarities and difference, and preliminarily summarizes genetic models of different types of uranium deposits. Finally, taking Erlian and Ordos basins as examples, the author makes an evaluation and a strategic analysis on the uranium metallogenic prospect of the above two basins

  17. Analysis on deep metallogenic trace and simulation experiment in xiangshan large-scale volcanic hydrothermal type uranium deposit

    International Nuclear Information System (INIS)

    Liu Zhengyi; Liu Zhangyue; Wen Zhijian; Du Letian

    2010-01-01

    Based on series experiments on field geologic analysis, and associated with deep metallogenic trace experiment model transformed from establishment of field deep metallogenic trace model, this paper come to the conclusion that distribution coefficients of U and Th first domestic from the magmatic experiment, and then discuss the geochemical behaviors of U, Th, K during magmatic evolution stage. The experiment shows that close relationship between U and Na during the hydrothermal alteration stage; and relationship between U and K during metallogenic stage, which prove that U and K are incompatible and regularity of variation between K and Na. The conclusion of uranium dissolving ability increased accompany with pressure increasing in basement metamorphic rocks and host rocks, is obtained from this experiment, which indicate a good deep metallogenic prospect. Furthermore, Pb, Sr, Nd, He isotopes show that the volcanic rocks and basement rocks are ore source beds; due to the combined functions of volcanic hydrothermal and mantle ichor, uranium undergo multi-migration and enrichment and finally concentrated to large rich deposit. (authors)

  18. Analysis of uranium metallogenic conditions and prospective prognosis on the south-central part of the Xikang-Yunnan Axis

    International Nuclear Information System (INIS)

    Guo Baochi; Qian Farong; Cai Yuqi; Zhang Daishi

    1996-08-01

    Three types of uranium mineralizations (i.e., sandstone type, sodium metasomatite type and Proterozoic epimetamorphic rock type) are distinguished in the south-central part of the Xikang-Yunnan Axis, and their major characteristics are expounded. It is proposed that the Early Proterozoic crust has three elevation stages: primitive old land formation stage, subsidence stage and base mental reconsolidation stage, and that the Middle Proterozoic Kunyang Group is different from typical carbonate-siliceous-politic sedimentary formation which contains many regional uranium-rich horizons. The primitive uranium contents of some rocks in the region have been studied for the first time, and proposed a new knowledge that some relative rocks be recognized as the uranium source rocks of the region. It is pointed out that there are two kinds of hydrothermal alterations, namely, alkaline alteration and acidic alteration, and some areas with development of alkaline alteration have promising uranium-metallogenic potential. As for the U-metallogenic prospect of the region, three conclusions are summarized: (1) This region does not have favourable geologic conditions for the Australian-Canadian type Proterozoic unconformity-related uranium deposit; (2) The Proterozoic unconformity different from that of Australian-Canadian type does not have permission U-metallogenic potential either; (3) The alkaline (sodium) metasomatite type uranium mineralization in the region has some prospecting potential. Therefore on the basis of above-mentioned conclusions five relatively promising uranium-metallogenic prospects are selected. (4 refs.)

  19. Temporal evolution of granitic magmas in the Luanchuan metallogenic belt, east Qinling Orogen, central China: Implications for Mo metallogenesis

    Science.gov (United States)

    Li, Dong; Han, Jiangwei; Zhang, Shouting; Yan, Changhai; Cao, Huawen; Song, Yaowu

    2015-11-01

    The Luanchuan metallogenic belt, located within the eastern part of the Qinling Orogen, central China, hosts a number of world-class Mo deposits that are closely related to small late Mesozoic granitic plutons. Zircon U-Pb dating of distinct plutons in the Luanchuan metallogenic belt has yielded ages of 153 ± 1, 154 ± 2, 152 ± 2, and 148 ± 1 Ma. Molybdenite Re-Os isotopic compositions of Yuku ore district in the southern part of Luanchuan metallogenic belt has yielded an isochron age of 146 ± 1 Ma, which is consistent with the large-scale mineralization ages in the northern part of the Luanchuan metallogenic belt. A combination of previous studies and new geochronological and isotopic data show a concordant temporal and genetic link between granitic magmatism and Mo mineralization in the Luanchuan metallogenic belt, suggesting that this mineralization episode formed the most extensive Mo mineralization belt in the east Qinling Orogen. Zircon grains from Mo-related granitic plutons show similar trace element distributions. High-precision Multi Collector-Inductively Coupled Plasma-Mass Spectrometry (MC-ICP-MS) Pb isotope analysis of K-feldspar megacrysts from mineralization-related granites suggest that they were derived from the lower crust. Similarly, the Pb isotopic compositions of pyrite coprecipitated with molybdenite also suggest that the metals were derived form the lower crust, with probably minor mantle contribution. A continuum mineralization model that describes the sourcing of Mo from an evolving granitic magma over successive differentiation events, possibly in separate but connected magma chambers, could explain the remarkable Mo enrichment in the Luanchuan metallogenic belt. The volatile- and Mo-bearing granitic magmas ascended as diapirs from the deep crust, and were emplaced as dikes in the upper crust. Lithological differences between these Mo-bearing granites may relate to different stages in the evolution of individual magmas. Finally, ore

  20. Loss Factor Characterization Methodology for Piezoelectric Ceramics

    International Nuclear Information System (INIS)

    Zhuang Yuan; Ural, Seyit O; Uchino, Kenji

    2011-01-01

    The key factor for the miniaturization of piezoelectric devices is power density, which is limited by the heat generation or loss mechanisms. There are three loss components for piezoelectric vibrators, i.e., dielectric, elastic and piezoelectric losses. The mechanical quality factor, determined by these three factors, is the figure of merit in the sense of loss or heat generation. In this paper, quality factors of resonance and antiresonance for k 31 , k 33 , and k 15 vibration modes are derived, and the methodology to determine loss factors in various directions is provided. For simplicity, we focus on materials with ∞mm (equivalent to 6mm) crystal symmetry for deriving the loss factors of polycrystalline ceramics, and 16 different loss factors among total 20 can be obtained from the admittance/ impedance measurements.

  1. Magmatism and polymetallic mineralization in southwestern Qinzhou-Hangzhou metallogenic belt, South China

    Science.gov (United States)

    Huang, Xudong; Lu, Jianjun; Wang, Rucheng; Ma, Dongsheng

    2016-04-01

    As Neoproterozoic suture zone between the Yangtze Block and Cathaysia Block, Qinzhou-Hangzhou metallogenic belt is one of the 21 key metallogenic belts in China. Intensive multiple-aged felsic magmatism and related polymetallic mineralization take place in this belt. Although Neoproterozoic, Paleozoic, Triassic granites and associated deposits have been found in southwestern Qinzhou-Hangzhou metallogenic belt, Middle-Late Jurassic (150-165 Ma) magmatism and related mineralization is of the most importance. Three major kinds of Middle-Late Jurassic granitoids have been distinguished. (Cu)-Pb-Zn-bearing granitoids are slightly differentiated, calc-alkaline and metaluminous dioritic to granodioritic rocks. Sn-(W)-bearing granites contain dark microgranular enclaves and have high contents of REE and HFSE, suggesting affinities of aluminous A-type (A2) granites. W-bearing granites are highly differentiated and peraluminous rocks. (Cu)-Pb-Zn-bearing granitoids have ɛNd(t) values of -11 ˜ -4 and ɛHf(t) values of -12 ˜ -7, corresponding to TDMC(Nd) from 1.4 to 1.8 Ga and TDMC(Hf) from 1.6 to 2.0 Ga, respectively. The ɛNd(t) values of W-bearing granites vary from -11 to -8 with TDMC(Nd) of 1.6 ˜ 1.9 Ga and ɛHf(t) values change from -16 to -7 with TDMC(Hf) of 1.5 ˜ 2.0 Ga. Compared with (Cu)-Pb-Zn-bearing granitoids and W-bearing granites, the Sn-(W)-bearing granites have higher ɛNd(t) (-8 ˜ -2) and ɛHf(t) (-8 ˜ -2) values and younger TDMC(Nd) (1.1 ˜ 1.6 Ga) and TDMC(Hf) (1.2 ˜ 1.8 Ga) values, showing a more juvenile isotopic character. Sn-(W)-bearing granites originate from partial melting of granulitized lower crust involved with some mantle-derived materials. W-bearing granites are derived from partial melting of crust. (Cu)-Pb-Zn-bearing granitoids are also derived from crust but may be influenced by more mantle-derived materials. For (Cu)-Pb-Zn deposits, skarn and carbonate replacement are the most important mineralization types. Cu ore bodies mainly

  2. Nature, diversity of deposit types and metallogenic relations of South China

    Science.gov (United States)

    Zaw, K.; Peters, S.G.; Cromie, P.; Burrett, C.; Hou, Z.

    2007-01-01

    The South China Region is rich in mineral resources and has a wide diversity of deposit types. The region has undergone multiple tectonic and magmatic events and related metallogenic processes throughout the earth history. These tectonic and metallogenic processes were responsible for the formation of the diverse styles of base and precious metal deposits in South China making it one of the resource-rich regions in the world. During the Proterozoic, the South China Craton was characterised by rifting of continental margin before eruption of submarine volcanics and development of platform carbonate rocks, and the formation of VHMS, stratabound copper and MVT deposits. The Phanerozoic metallogeny of South China was related to opening and closing of the Tethyan Ocean involving multiple orogenies by subduction, back-arc rifting, arc-continent collision and post-collisional extension during the Indosinian (Triassic), Yanshanian (Jurassic to Cretaceous) and Himalayan (Tertiary) Orogenies. The Late Palaeozoic was a productive metallogenic period for South China resulting from break-up and rifting of Gondwana. Significant stratabound base and precious metal deposits were formed during the Devonian and Carboniferous (e.g., Fankou and Dabaoshan deposits). These Late Palaeozoic SEDEX-style deposits have been often overprinted by skarn systems associated with Yanshanian magmatism (e.g., Chengmenshan, Dongguashan and Qixiashan). A number of Late Palaeozoic to Early Mesozoic VHMS deposits also developed in the Sanjiang fold belt in the western part of South China (e.g., Laochang and Gacun). South China has significant sedimentary rock-hosted Carlin-like deposits, which occur in the Devonian- to Triassic-aged accretionary wedge or rift basins at the margin of the South China Craton. They are present in a region at the junction of Yunnan, Guizhou, and Guangxi Provinces called the 'Southern Golden Triangle', and are also present in NW Sichuan, Gansu and Shaanxi, in an area known as

  3. Hydrothermal Evolution of the Giant Cenozoic Kadjaran porphyry Cu-Mo deposit, Tethyan metallogenic belt, Armenia, Lesser Caucasus: mineral paragenetic, cathodoluminescence and fluid inclusion constraints

    Science.gov (United States)

    Hovakimyan, Samvel; Moritz, Robert; Tayan, Rodrik; Rezeau, Hervé

    2016-04-01

    The Lesser Caucasus belongs to the Central segment of the Tethyan metallogenic belt and it is a key area to understand the metallogenic evolution between the Western & Central parts of the Tethyan belt and its extension into Iran. Zangezur is the most important mineral district in the southernmost Lesser Caucasus. It is a component of the South Armenian block, and it was generated during the convergence and collision of the southern margin of the Eurasian plate and the northern margin of the Arabian plate, and terranes of Gondwana origin (Moritz et al., in press). The Zangezur ore district consists of the Tertiary Meghri-Ordubad composite pluton, which is characterized by a long-lasting Eocene to Pliocene magmatic, tectonic and metallogenic evolution. It hosts major porphyries Cu-Mo and epithermal Au - polymetallic deposits and occurrences, including the giant world class Kadjaran porphyry Cu-Mo deposit (2244 Mt reserves, 0.3% Cu, 0.05% Mo and 0.02 g/t Au). The Kadjaran deposit is hosted by a monzonite intrusion (31.83±0.02Ma; Moritz et al., in press). Detailed field studies of the porphyry stockwork and veins of the different mineralization stages, their crosscutting and displacement relationships and the age relationship between different paragenetic mineral associations were the criteria for distinction of the main stages of porphyry mineralization at the Kadjaran deposit. The economic stages being: quartz- molybdenite, quartz-molybdenite-chalcopyrite, and quartz-chalcopyrite. The main paragenetic association of the Kadjaran porphyry deposit includes pyrite, molybdenite, chalcopyrite, bornite, chalcocite, pyrrhotite, covellite, sphalerite, and galena. Recent field observations in the Kadjaran open pit revealed the presence of epithermal veins with late vuggy silica and advanced argillic alteration in the north-eastern and eastern parts of the deposit. They are distributed as separate veins and have also been recognized in re-opened porphyry veins and in

  4. Research on metallogenic conditions of intersection-type uranium ore-deposits in Zhongdong area, Northern Guangdong province

    International Nuclear Information System (INIS)

    Wang Zhengqi; Wu Lieqin; Zhang Guoyu

    2007-12-01

    The methods following as field geological investigation, trace element geo- chemistry and isotope geochemistry were used in this project. Based on geological and geochemical characteristics of Xiaoshui uranium ore deposits in Zhongdong area, Xiazhuang ore-field, Guangdong province, it could be concluded that: (1) The Provenance of Cretaceous mantle is a enriched mantle; (2) Silicified zone-type and intersection-type uranium ore are distinctness in the metallogenic period and mineralization process, and main metallogenic period of Xiaoshui uranium ore-deposit is 73.5 Ma; (3) The sources of uranium mineralization substance derived from enriched mantle; and (4)The intersection-type high grade uranium deposits were controlled by substances derived from mantle (contain with U, CO 2 , F, et al), tracks of intersection of NWW-across with NNE-trending faults and lithology of diabase. (authors)

  5. Research on metallogenic conditions of intersection-type uranium ore-deposits in Zhongdong area, Northern Guangdong province

    Energy Technology Data Exchange (ETDEWEB)

    Zhengqi, Wang [East China Inst. of Technology, Fuzhou (China); [Beijing Research Inst. of Uranium Geology, Beijing (China); Lieqin, Wu [Institute No.290, CNNC, Shaoguan (China); Guoyu, Zhang [East China Inst. of Technology, Fuzhou (China)

    2007-12-15

    The methods following as field geological investigation, trace element geo- chemistry and isotope geochemistry were used in this project. Based on geological and geochemical characteristics of Xiaoshui uranium ore deposits in Zhongdong area, Xiazhuang ore-field, Guangdong province, it could be concluded that: (1) The Provenance of Cretaceous mantle is a enriched mantle; (2) Silicified zone-type and intersection-type uranium ore are distinctness in the metallogenic period and mineralization process, and main metallogenic period of Xiaoshui uranium ore-deposit is 73.5 Ma; (3) The sources of uranium mineralization substance derived from enriched mantle; and (4)The intersection-type high grade uranium deposits were controlled by substances derived from mantle (contain with U, CO{sub 2}, F, et al), tracks of intersection of NWW-across with NNE-trending faults and lithology of diabase. (authors)

  6. Acute otitis media: characterization and risk factors

    Directory of Open Access Journals (Sweden)

    Amalia Nieves Álvarez

    2011-03-01

    Full Text Available La otitis media aguda es una de las infecciones respiratorias superiores más frecuentes en la infancia, fundamentalmente en niños menores de seis años. Su elevada incidencia y recidiva se deben a la interacción entre diferentes factores de riesgo. El signo más específico de presentación es la otalgia, acompañándose de fiebre, malestar y disminución de la audición.Por la frecuencia de niños con otitis media aguda en las consultas externas y de urgencia del hospital, nos planteamos conocer su comportamiento en el grupo estudiado. Se realizó un estudio descriptivo longitudinal retrospectivo a los pacientes que acudieron a consulta de Otorrinolaringología con otitis media aguda del Hospital Pediátrico Universitario Centro Habana, ubicado en La Habana, en el período comprendido entre enero de 2006 hasta diciembre de 2009. Se caracterizaron los pacientes en cuanto a edad, sexo, factores predisponentes, manifestaciones clínicas, etapa clínica de la otitis, recurrencia y tratamiento. Resultados: El sexo masculino y los menores de cinco años fueron los más afectados. Los factores de riesgo predominantes fueron el tabaquismo pasivo, asistencia a círculos infantiles y lactancia materna insuficiente. Las manifestaciones clínicas más frecuentes fueron los síntomas catarrales, otalgia y fiebre. La otitis media con trasudado fue la etapa clínica más observada. El grupo de edades que mayor recurrencia presentó fue el de uno a cuatro años y la terapia antimicrobiana con amoxicilina fue la más utilizada.

  7. Re–Os geochronology of Cu and W–Mo deposits in the Balkhash metallogenic belt, Kazakhstan and its geological significance

    Directory of Open Access Journals (Sweden)

    Xuanhua Chen

    2010-10-01

    Full Text Available The Central Asian metallogenic domain (CAMD is a multi-core metallogenic system controlled by boundary strike-slip fault systems. The Balkhash metallogenic belt in Kazakhstan, in which occur many large and super-large porphyritic Cu–Mo deposits and some quartz vein- and greisen-type W–Mo deposits, is a well-known porphyritic Cu–Mo metallogenic belt in the CAMD. In this paper 11 molybdenite samples from the western segment of the Balkhash metallogenic belt are selected for Re–Os compositional analyses and Re–Os isotopic dating. Molybdenites from the Borly porphyry Cu deposit and the three quartz vein-greisen W–Mo deposits—East Kounrad, Akshatau and Zhanet—all have relatively high Re contents (2712–2772 μg/g for Borly and 2.267–31.50 μg/g for the other three W–Mo deposits, and lower common Os contents (0.670–2.696 ng/g for Borly and 0.0051–0.056 ng/g for the other three. The molybdenites from the Borly porphyry Cu–Mo deposit and the East Kounrad, Zhanet, and Akshatau quartz vein- and greisen-type W–Mo deposits give average model Re–Os ages of 315.9 Ma, 298.0 Ma, 295.0 Ma, and 289.3 Ma respectively. Meanwhile, molybdenites from the East Kounrad, Zhanet, and Akshatau W–Mo deposits give a Re–Os isochron age of 297.9 Ma, with an MSWD value of 0.97. Re–Os dating of the molybdenites indicates that Cu–W–Mo metallogenesis in the western Balkhash metallogenic belt occurred during Late Carboniferous to Early Permian (315.9–289.3 Ma, while the porphyry Cu–Mo deposits formed at ∼316 Ma, and the quartz vein-greisen W–Mo deposits formed at ∼298 Ma. The Re–Os model and isochron ages thus suggest that Late Carboniferous porphyry granitoid and pegmatite magmatism took place during the late Hercynian movement. Compared to the Junggar-East Tianshan porphyry Cu metallogenic belt in northwestern China, the formation of the Cu–Mo metallogenesis in the Balkhash metallogenic belt occurred between that of

  8. Uranium mineralization environment and prospecting potential of Dawan ore field in Nanling metallogenic belt

    International Nuclear Information System (INIS)

    Yang Shanghai

    2011-01-01

    Located in the middle part of Jiuyishan complex pluton, Nanling metallogenic belt, Dawan uranium ore field in Hunan Province is an important uranium-producing and rare metal, nonferrous metal cluster area due to the favourable mineralization environment. The Cambrian is the main uranium source bed and their contact zone to the pluton is the favorable part for mineralization. The uranium deposits which have been explored are all located in the exocontact zone of Jinjiling pluton in the middle part of Jiuyishan complex pluton which is composed of the independent eastern and western magma evolution centers. In the west center, Jinjiling pluton is closely related to uranium mineralization where the trinity geologic setting was formed with magma evolution, hydrothermal fluid action and mineralization. The deep slitted and large faults provide the pathway and thermodynamic source for circulating migration of mineralizing fluid. The uranium mineralization mainly occurred in crustal stress conversion period of Late Cretaceous and related to the tensive NW extending fault and deep originated fluid. The gravity, aero magnetic, airborne gamma-ray spectrometry anomalies and radioactivity hydrochemical anomaly are important criteria for uranium prospecting. Based on the analysis of regional uranium mineralization environment, the prospecting potential is forecasted. (authors)

  9. Metallogenic model for continental volcanic-type rich and large uranium deposits

    International Nuclear Information System (INIS)

    Chen Guihua

    1998-01-01

    A metallogenic model for continental volcanic-type rich and large/super large uranium deposits has been established on the basis of analysis of occurrence features and ore-forming mechanism of some continental volcanic-type rich and large/super large uranium deposits in the world. The model proposes that uranium-enriched granite or granitic basement is the foundation, premetallogenic polycyclic and multistage volcanic eruptions are prerequisites, intense tectonic-extensional environment is the key for the ore formation, and relatively enclosed geologic setting is the reliable protection condition of the deposit. By using the model the author explains the occurrence regularities of some rich and large/super large uranium deposits such as Strelichof uranium deposit in Russia, Dornot uranium deposit in Mongolia, Olympic Dam Cu-U-Au-REE deposit in Australia, uranium deposit No.460 and Zhoujiashan uranium deposit in China, and then compares the above deposits with a large poor uranium deposit No.661 as well

  10. Characterization factors for thermal pollution in freshwater aquatic environments.

    Science.gov (United States)

    Verones, Francesca; Hanafiah, Marlia Mohd; Pfister, Stephan; Huijbregts, Mark A J; Pelletier, Gregory J; Koehler, Annette

    2010-12-15

    To date the impact of thermal emissions has not been addressed in life cycle assessment despite the narrow thermal tolerance of most aquatic species. A method to derive characterization factors for the impact of cooling water discharges on aquatic ecosystems was developed which uses space and time explicit integration of fate and effects of water temperature changes. The fate factor is calculated with a 1-dimensional steady-state model and reflects the residence time of heat emissions in the river. The effect factor specifies the loss of species diversity per unit of temperature increase and is based on a species sensitivity distribution of temperature tolerance intervals for various aquatic species. As an example, time explicit characterization factors were calculated for the cooling water discharge of a nuclear power plant in Switzerland, quantifying the impact on aquatic ecosystems of the rivers Aare and Rhine. The relative importance of the impact of these cooling water discharges was compared with other impacts in life cycle assessment. We found that thermal emissions are relevant for aquatic ecosystems compared to other stressors, such as chemicals and nutrients. For the case of nuclear electricity investigated, thermal emissions contribute between 3% and over 90% to Ecosystem Quality damage.

  11. Geochronology and geochemistry of the Badaguan porphyry Cu-Mo deposit in Derbugan metallogenic belt of the NE China, and their geological significances

    Science.gov (United States)

    Gao, Bingyu; Zhang, Lianchang; Jin, Xindi; Li, Wenjun; Chen, Zhiguang; Zhu, Mingtian

    2016-03-01

    The Badaguan porphyry Cu-Mo deposit belongs to the Derbugan metallogenic belt, which is located in the Ergun block, NE China. In the mining area, the Cu-Mo mineralization mainly occurs in quartz diorite porphyry and is hosted within silicified-sericitized and sericite alteration zone. Geochemical results of the host porphyry is characterized by high SiO2, high Al2O3, low MgO, weak positive Eu anomalies and clearly HREE depletion, high Sr, low Y and low Yb, similar to those of adakite. The Sr-Nd isotopic composition of the host porphyry displays an initial (87Sr/86Sr)i ratio of 0.7036-0.7055 and positive Nd( t) values of +0.1 to +0.6, which are similar to the OIB, reflecting the source of the host porphyry may derive from subducted ocean slab, and the new lower crust also had some contribution to the magma sources. The SIMS zircon U-Pb age from the host porphyry is 229 ± 2 Ma. The Re-Os isochron age for the molybdenite in the deposit is 225 ± 2 Ma closed to zircon U-Pb age of the host porphyry, indicating that Cu-Mo mineralization event occurred in Triassic. Combining the geology-geochemistry of the host porphyry and the regional tectonic evolution, we infer that the subduction processes of Mongol-Okhotsk oceanic slab under the Ergun block led to the formation of the Badaguan porphyry Cu-Mo deposit during the Triassic.

  12. Characterizing the transcriptome upon depletion of RNA processing factors

    DEFF Research Database (Denmark)

    Herudek, Jan

    , it is not clear how they target and discriminate their RNA substrates. Moreover, many novel RNA species are poorly characterized and their function is not understood. Over the last decade, protein function has been studied using RNA interference. However, this approach does not allow investigation of instant......The human genome is pervasively transcribed and produces an enormous amount of non-coding RNA (ncRNA). Compared to protein-coding transcripts, many classes of ncRNAs are very unstable and rapidly degraded by the RNA decay machinery. The RNA exosome complex is a main RNA ‘degrader’ in the human...... nucleus and is responsible for the proper processing and decay of a wide range of RNA molecules. Notably, the RNA exosome complex associates with a plethora of co-factors and activators that assist in the recognition of specific RNA substrates. Although many exosome partners have been characterized...

  13. Characterization of a human antigen specific helper factor

    International Nuclear Information System (INIS)

    Richardson, B.

    1986-01-01

    While antigen (Ag) specific helper factors have been characterized in mice, similar molecules have not been identified in humans. To characterize human antigen specific helper molecules, an IL-2 dependent tetanus toxoid (T.T.) reactive T cell line was fused with a 6-thioguanine resistant CEM line, and hybrids selected in medium containing hypoxanthine and azaserine. Hybrids were screened by culturing the cells with 35 S-Met then reacting the supernatants with T.T. or hepatitis vaccine immobilized on nitrocellulose. One hybrid, TT6BA-O, was identified which secreted a Met-containing molecule which bound T.T. but not hepatitis vaccine. Supernatants from TT6BA-O, but not the parent CEM line, when added to autologous peripheral blood mononuclear cells (PBMC's) stimulated secretion of T.T. specific antibodies (Abs). Specificity controls demonstrated that TT6BA-O supernatant did not induce antibodies to diphtheria toxoid, hepatitis vaccine or pneumococcal polysaccharide, and total immunoglobulin (lg) synthesis was minimally increased. In contrast, pokeweed mitogen stimulated significant lg synthesis as well as Ab's to pneumococcal polysaccharide and T.T. TT6BA-O supernatant induced anti-T.T.Ab's in autologous PBMC's but not PBMC's from 3 unrelated donors, suggesting that the activity of the helper factor is restricted, possibly by the MHC. The molecular weight of the helper factor was estimated at 100,000-150,000 by Sephacryl S-300 chromatography. Finally, the helper factor could be demonstrated to bind and elute from sephorose-immobilized T.T. and anti-DR antisera, but not anti-lg antisera or the T40/25 monoclonal antibody, which binds a nonpolymorphic determinant on the human T cell receptor. These results demonstrate that human Ag specific helper factors exist, bind antigen and bear class II MHC determinants

  14. Factors Influencing the Phenotypic Characterization of the Oral Marker, PROP

    Directory of Open Access Journals (Sweden)

    Beverly J. Tepper

    2017-11-01

    Full Text Available In the last several decades, the genetic ability to taste the bitter compound, 6-n-propyltiouracil (PROP has attracted considerable attention as a model for understanding individual differences in taste perception, and as an oral marker for food preferences and eating behavior that ultimately impacts nutritional status and health. However, some studies do not support this role. This review describes common factors that can influence the characterization of this phenotype including: (1 changes in taste sensitivity with increasing age; (2 gender differences in taste perception; and (3 effects of smoking and obesity. We suggest that attention to these factors during PROP screening could strengthen the associations between this phenotype and a variety of health outcomes ranging from variation in body composition to oral health and cancer risk.

  15. Analysis of metallogenic conditions of sandstone type uranium deposits in interlayer oxidation zone in the northwest of Junggar basin, Xinjiang

    International Nuclear Information System (INIS)

    Lin Shuangxing

    1997-01-01

    From various aspects such as the basin structures, structure features of the sedimentary cover, lithofacies features of basin sediments, geochemical characteristics of sedimentary rocks, regional hydrogeologic conditions and epigenetic reworking of the sedimentary cover and so on, the author analyzes the metallogenic conditions of sandstone type uranium deposits in interlayer oxidation zone in the northwest of Junggar basin. The author proposes that the area has abundant uranium sources, and possesses favourable stratigraphic combinations for the development of interlayer oxidation zone. Secondarily oxidized sandstone bodies present universally at places from the source area to stream channel facies. Reducing beds and secondarily reducing barrier are developed at the front of the secondarily oxidized sandstone body. The tectonic features of the sedimentary cover indicate that the area belongs to a relatively-stable suborogenic region and possesses secondary mobilization and reworking conditions during the uplifting and contraction stage of the basin. Epigenetic metallization is evident in the sedimentary cover of the basin

  16. Characterization of five partial deletions of the factor VIII gene

    International Nuclear Information System (INIS)

    Youssoufian, H.; Antonarakis, S.E.; Aronis, S.; Tsiftis, G.; Phillips, D.G.; Kazazian, H.H. Jr.

    1987-01-01

    Hemophilia A is an X-linked disorder of coagulation caused by a deficiency of factor VIII. By using cloned DNA probes, the authors have characterized the following five different partial deletions of the factor VIII gene from a panel of 83 patients with hemophilia A: (i) a 7-kilobase (kb) deletion that eliminates exon 6; (ii) a 2.5-kb deletion that eliminates 5' sequences of exon 14; (iii) a deletion of at least 7 kb that eliminates exons 24 and 25; (iv) a deletion of at least 16 kb that eliminates exons 23-25; and (v) a 5.5-kb deletion that eliminates exon 22. The first four deletions are associated with severe hemophilia A. By contrast, the last deletion is associated with moderate disease, possibly because of in-frame splicing from adjacent exons. None of those patients with partial gene deletions had circulating inhibitors to factor VIII. One deletion occurred de novo in a germ cell of the maternal grandmother, while a second deletion occurred in a germ cell of the maternal grandfather. These observations demonstrate that de novo deletions of X-linked genes can occur in either male or female gametes

  17. Characterization of a diffusible signaling factor from Xylella fastidiosa.

    Science.gov (United States)

    Beaulieu, Ellen D; Ionescu, Michael; Chatterjee, Subhadeep; Yokota, Kenji; Trauner, Dirk; Lindow, Steven

    2013-01-08

    Cell-cell signaling in Xylella fastidiosa has been implicated in the coordination of traits enabling colonization in plant hosts as well as insect vectors. This cell density-dependent signaling has been attributed to a diffusible signaling factor (DSF) produced by the DSF synthase RpfF. DSF produced by related bacterial species are unsaturated fatty acids, but that of X. fastidiosa was thought to be different from those of other taxa. We describe here the isolation and characterization of an X. fastidiosa DSF (XfDSF) as 2(Z)-tetradecenoic acid. This compound was isolated both from recombinant Erwinia herbicola expressing X. fastidiosa rpfF and from an X. fastidiosa rpfC deletion mutant that overproduces DSF. Since an rpfF mutant is impaired in biofilm formation and underexpresses the hemagglutinin-like protein-encoding genes hxfA and hxfB, we demonstrate that these traits can be restored by ca. 0.5 µM XfDSF but not by myristic acid, the fully saturated tetradecenoic acid. A phoA-based X. fastidiosa biosensor that assesses DSF-dependent expression of hxfA or hxfB revealed a high level of molecular specificity of DSF signaling. X. fastidiosa causes diseases in many important plants, including grape, where it incites Pierce's disease. Virulence of X. fastidiosa for grape is coordinated by cell-cell signaling molecules, designated DSF (Diffusible Signaling Factor). Mutants blocked in DSF production are hypervirulent for grape, suggesting that virulence is suppressed upon DSF accumulation and that disease could be controlled by artificial elevation of the DSF level in plants. In this work, we describe the isolation of the DSF produced by X. fastidiosa and the verification of its biological activity as an antivirulence factor. We also have developed X. fastidiosa DSF biosensors to evaluate the specificity of cell-cell signaling to be investigated.

  18. Complexity and Productivity Differentiation Models of Metallogenic Indicator Elements in Rocks and Supergene Media Around Daijiazhuang Pb–Zn Deposit in Dangchang County, Gansu Province

    International Nuclear Information System (INIS)

    He, Jin-zhong; Yao, Shu-zhen; Zhang, Zhong-ping; You, Guan-jin

    2013-01-01

    With the help of complexity indices, we quantitatively studied multifractals, frequency distributions, and linear and nonlinear characteristics of geochemical data for exploration of the Daijiazhuang Pb–Zn deposit. Furthermore, we derived productivity differentiation models of elements from thermodynamics and self-organized criticality of metallogenic systems. With respect to frequency distributions and multifractals, only Zn in rocks and most elements except Sb in secondary media, which had been derived mainly from weathering and alluviation, exhibit nonlinear distributions. The relations of productivity to concentrations of metallogenic elements and paragenic elements in rocks and those of elements strongly leached in secondary media can be seen as linear addition of exponential functions with a characteristic weak chaos. The relations of associated elements such as Mo, Sb, and Hg in rocks and other elements in secondary media can be expressed as an exponential function, and the relations of one-phase self-organized geological or metallogenic processes can be represented by a power function, each representing secondary chaos or strong chaos. For secondary media, exploration data of most elements should be processed using nonlinear mathematical methods or should be transformed to linear distributions before processing using linear mathematical methods.

  19. Development and characterization of recombinant ovine coagulation factor VIII.

    Directory of Open Access Journals (Sweden)

    Philip M Zakas

    Full Text Available Animal models of the bleeding disorder, hemophilia A, have been an integral component of the biopharmaceutical development process and have facilitated the development of recombinant coagulation factor VIII (fVIII products capable of restoring median survival of persons with hemophilia A to that of the general population. However, there remain several limitations to recombinant fVIII as a biotherapeutic, including invasiveness of intravenous infusion, short half-life, immunogenicity, and lack of availability to the majority of the world's population. The recently described ovine model of hemophilia A is the largest and most accurate phenocopy. Affected sheep die prematurely due to bleeding-related pathogenesis and display robust adaptive humoral immunity to non-ovine fVIII. Herein, we describe the development and characterization of recombinant ovine fVIII (ofVIII to support further the utility of the ovine hemophilia A model. Full-length and B-domain deleted (BDD ofVIII cDNAs were generated and demonstrated to facilitate greater biosynthetic rates than their human fVIII counterparts while both BDD constructs showed greater expression rates than the same-species full-length versions. A top recombinant BDD ofVIII producing baby hamster kidney clone was identified and used to biosynthesize raw material for purification and biochemical characterization. Highly purified recombinant BDD ofVIII preparations possess a specific activity nearly 2-fold higher than recombinant BDD human fVIII and display a differential glycosylation pattern. However, binding to the carrier protein, von Willebrand factor, which is critical for stability of fVIII in circulation, is indistinguishable. Decay of thrombin-activated ofVIIIa is 2-fold slower than human fVIII indicating greater intrinsic stability. Furthermore, intravenous administration of ofVIII effectively reverses the bleeding phenotype in the murine model of hemophilia A. Recombinant ofVIII should facilitate

  20. Medication Desensitization: Characterization of Outcomes and Risk Factors for Reactions.

    Science.gov (United States)

    Murray, Taryn S; Rice, Todd W; Wheeler, Arthur P; Phillips, Elizabeth J; Dworski, Ryszard T; Stollings, Joanna L

    2016-03-01

    Although its mechanisms are poorly understood, desensitization has been used to induce a temporary state of immune unresponsiveness in patients who have IgE-, non-IgE-, or pharmacologically mediated reactions when a drug has no alternatives. The purpose of this study was to characterize the outcomes and identify risk factors for reactions during drug desensitization. A retrospective review of electronic medical records of adult patients undergoing drug desensitization from January 1, 2011, to December 31, 2013, was conducted in 2 intensive care units at a tertiary medical center. We used multivariate analysis to determine if specified risk factors were associated with reacting during the desensitization. Reactions were classified according to the pretest probability prior to desensitization, and then, reactions during desensitization were classified based on the occurrence of cutaneous reactions as follows: successful with no reaction, mild reaction, moderate reaction, or failed. Failure could result from any systemic allergic or cutaneous reaction resulting in procedure termination. The desensitizations were also assessed to determine if the patient required de-escalation secondary to a reaction. A total of 88 desensitizations were performed in 69 patients. Desensitization was completed with no cutaneous reaction in 85% of patients. No baseline characteristic, medication class (P = 0.46), or indication for desensitization (P = 0.59) was associated with having a reaction. Reported histories of urticaria (P desensitization. However, neither history of urticaria nor labored breathing was independently associated with having a reaction in multivariate analysis (OR = 0.979, 95% CI = 0.325-2.952, P = 0.970, and OR = 1.626, 95% CI = 0.536-4.931, P = 0.739, respectively). Drug desensitization is safe for patients who have no alternative for therapy. Reported allergy histories of urticaria and labored breathing are both associated with having a reaction during the

  1. Characterizing risk factors for pediatric lamp oil product exposures.

    Science.gov (United States)

    Sheikh, S; Chang, A; Kieszak, S; Law, R; Bennett, H K W; Ernst, E; Bond, G R; Spiller, H A; Schurz-Rogers, H; Chu, A; Bronstein, A C; Schier, J G

    2013-11-01

    Poisonings from lamp oil ingestion continue to occur worldwide among the pediatric population despite preventive measures such as restricted sale of colored and scented lamp oils. This suggests that optimal prevention practices for unintentional pediatric exposures to lamp oil have yet to be identified and/or properly implemented. To characterize demographic, health data, and potential risk factors associated with reported exposures to lamp oil by callers to poison centers (PCs) in the US and discuss their public health implications. This was a two part study in which the first part included characterizing all exposures to a lamp oil product reported to the National Poison Data System (NPDS) with regard to demographics, exposure, health, and outcome data from 1/1/2000 to 12/31/2010. Regional penetrance was calculated using NPDS data by grouping states into four regions and dividing the number of exposure calls by pediatric population per region (from the 2000 US census). Temporal analyses were performed on NPDS data by comparing number of exposures by season and around the July 4th holiday. Poisson regression was used to model the count of exposures for these analyses. In the second part of this project, in order to identify risk factors we conducted a telephone-based survey to the parents of children from five PCs in five different states. The 10 most recent lamp oil product exposure calls for each poison center were systematically selected for inclusion. Calls in which a parent or guardian witnessed a pediatric lamp oil product ingestion were eligible for inclusion. Data on demographics, exposure information, behavioral traits, and health were collected. A descriptive analysis was performed and Fisher's exact test was used to evaluate associations between variables. All analyses were conducted using SAS v9.3. Among NPDS data, 2 years was the most common patient age reported and states in the Midwestern region had the highest numbers of exposure calls compared to

  2. Environmental risk assessment of lead-zinc mining: a case study of Adudu metallogenic province, middle Benue Trough, Nigeria.

    Science.gov (United States)

    Igwe, Ogbonnaya; Una, Chuku Okoro; Abu, Ezekiel; Adepehin, Ekundayo Joseph

    2017-09-07

    Assessment of the impacts of lead-zinc mining in Adudu-Imon metallogenic province was carried out. Reconnaissance and detailed field studies were done. Lithologies, stream sediments, farmland soils, mine tailings, artificial pond water, stream water, well water, and borehole water were collected and subjected to atomic absorption spectrometry (AAS) and X-ray fluorescence (XRF) analyses. Geochemical maps were generated using ArcGIS 10.1. Significant contamination with cadmium (Cd), iron (Fe), and lead (Pb) was recorded in the collected water samples. Virtually all collected soil samples were observed to be highly contaminated when compared with the European Union environmental policy standard. The discharge of mining effluents through farmlands to the Bakebu stream, which drains the area, further exposes the dwellers of this environment to the accumulation of potentially harmful metals (PHMs) in their bodies through the consumption of food crops, aquatic animals, and domestic uses of the water collected from the stream channels. The study revealed non-conformity of past mining operations in the Adudu-Imon province to existing mining laws in Nigeria. Inhabitants of this region should stop farming in the vicinity of the mines, fishing from the Bakebu stream channels should be discouraged, and domestic use of the water should be condemned, even as concerned government agencies put necessary mercenaries in place to ensure conformity of miners to standard mining regulations in Nigeria.

  3. The lithosphere structure and deep processes of the Mesozoic metallogenic belt in eastern China: constraints from passive and active seismic methods

    Science.gov (United States)

    Lu, Q.; Shi, D.; Jiang, G.; Yan, J.

    2013-12-01

    The lithosphere structure and deep processes are keys to understanding mineral system and ore-forming processes. Lithosphere-scale process could create big footprints or signatures which can be observed by geophysics methods. SinoProbe-03 has conducted a Transect exploration across middle and lower Yangtze Metallogenic Belt (YMT) in Eastern China. Broadband seismic, reflection seismic, wide-angle reflection and magnetotellurics survey were carried out along the Transect. Seismic reflection profiles and MT survey were also performed in Luzong, Tongling and Ningwu ore districts to construct 3D geological model. The resulting geophysical data provides new information which help to better understanding the lithosphere structure, deep processes and deformation history of the Metallogenic Belt. The major results are: (1) Lower velocity body at the top of upper mantle and a SE dipping high velocity body were imaged by teleseismic tomography beneath YMB; (2) Shear wave splitting results show NE parallel fast-wave polarization direction which parallel with tectonic lineament; (3) The reflection seismic data support the crustal-detachment model, the lower and upper crust was detached during contraction deformation near Tanlu fault and Ningwu volcanic basin; (4) Broadband and reflection seismic confirm the shallow Moho beneath YMB; (5) Strong correlation of lower crust reflectivity with magmatism; (6) The lower crust below Luzong Volcanics shows obvious reflective anisotropy both at the crust-mantle transition and the brittle-ductile transition in the crust. All these features suggest that introcontinental subduction, lithosphere delamination, mantle sources magmatic underplating, and MASH process are responsible for the formation of this Mesozoic metallogenic belt. Acknowledgment: We acknowledge the financial support of SinoProbe by the Ministry of Finance and Ministry of Land and Resources, P. R. China, under Grant sinoprobe-03, and financial support by National Natural

  4. The lithosphere architecture and geodynamic of the Middle and Lower Yangtze metallogenic belt in eastern China: constraints from integrated geophysical data

    Science.gov (United States)

    Lü, Qingtian; Shi, Danian; Jiang, Guoming; Dong, Shuwen

    2014-05-01

    The lithosphere structure and deep processes are keys to understanding mineral system and ore-forming processes. Lithosphere-scale process could create big footprints or signatures which can be observed by geophysics methods. SinoProbe has conducted an integrated deep exploration across middle and lower reaches of Yangtze Metallogenic Belt (YMB) in Eastern China, these included broadband seismic, reflection seismic, wide-angle reflection and magnetotellurics survey. Seismic reflection profiles and MT survey were also performed in Luzong, Tongling and Ningwu ore districts to construct 3D geological model. The resulting geophysical data provides new information which help to better understanding the lithosphere structure, geodynamic, deformation and heat and mass transportation that lead to the formation of the Metallogenic Belt. The major results are: (1) Lower velocity body at the top of upper mantle and a SE dipping high velocity body were imaged by teleseismic tomography beneath YMB; (2) Shear wave splitting results show NE parallel fast-wave polarization direction which parallel with tectonic lineament; (3) The reflection seismic data support the crustal-detachment model, the lower and upper crust was detached during contraction deformation near Tanlu fault and Ningwu volcanic basin; (4) Broadband and reflection seismic confirm the shallow Moho beneath YMB; (5) Strong correlation of lower crust reflectivity with magmatism; (6) The lower crust below Luzong Volcanics shows obvious reflective anisotropy both at the crust-mantle transition and the brittle-ductile transition in the crust. All these features suggest that introcontinental subduction, lithosphere delamination, mantle sources magmatic underplating, and MASH process are responsible for the formation of this Mesozoic metallogenic belt. Acknowledgment: We acknowledge the financial support of SinoProbe by the Ministry of Finance and Ministry of Land and Resources, P. R. China, under Grant sinoprobe-03, and

  5. Perspective and resource evaluation and metallogenic studies on sandstone-type uranium deposit in Qianjiadian depression of Songliao Basin

    Energy Technology Data Exchange (ETDEWEB)

    Yuliang, Xia; Jinrong, Lin; Ziying, Li; Shengxiang, Li; Hanbin, Liu; Zhiming, Wang; Guang, Fan [Beijing Research Inst. of Uranium Geology, Beijing (China); Jiwei, Zheng; Zhenji, Li; Mingyu, Zhang [Liaohe Oil Field, Panjin (China)

    2003-07-01

    The geotectonic evolution history of the southeastern part of Songliao Basin has been clearly described and it is pointed out that both of the provenance rocks and evolution features of the studied area are favorable to the formation of U-rich sandstone bodies, development of interlayered oxidation and providing uranium source for mineralization. Yaojia Formation in Qianjiadian depression has been found out to be the favorable target ore bed for looking for sandstone-type uranium deposit. On the basis of analysis of metallogenetic conditions, the perspective target area has been circled and a sandstone-type uranium deposit with a certain amounts of uranium tonnages has been discovered. The achievements and data have been gotten in the following aspects: constitution and features of ore-forming beds and sandstone bodies, uranium existence forms and mineralogical and chemical compositions of the ores, associated elements and their economic values for comprehensive mining. The study of metallogenetic features and mechanism of the uranium deposit suggested that pre-enrichment of uranium during the depositional-diagenetic stage provide a good basis for uranium mineralization, and hereafter interlayered oxidation as well as oil-gas reduction processes played a decisive role to uranium mineralization. U-Pb isotopic studies indicate that the ores have two isochron ages of 53{+-}3 Ma and 7.0{+-}0 Ma, corresponding to the periods of arid and semiarid paleo-climates which are favorable to interlayered oxidation development and uranium mineralization. It is concluded that the Qianjiadian sandstone-type uranium deposit is genetically related to interlayered oxidation and secondary reduction of oil-gas. The metallogenic model of Qianjiadian uranium deposit was set up. (authors)

  6. Perspective and resource evaluation and metallogenic studies on sandstone-type uranium deposit in Qianjiadian depression of Songliao Basin

    International Nuclear Information System (INIS)

    Xia Yuliang; Lin Jinrong; Li Ziying; Li Shengxiang; Liu Hanbin; Wang Zhiming; Fan Guang; Zheng Jiwei; Li Zhenji; Zhang Mingyu

    2003-01-01

    The geotectonic evolution history of the southeastern part of Songliao Basin has been clearly described and it is pointed out that both of the provenance rocks and evolution features of the studied area are favorable to the formation of U-rich sandstone bodies, development of interlayered oxidation and providing uranium source for mineralization. Yaojia Formation in Qianjiadian depression has been found out to be the favorable target ore bed for looking for sandstone-type uranium deposit. On the basis of analysis of metallogenetic conditions, the perspective target area has been circled and a sandstone-type uranium deposit with a certain amounts of uranium tonnages has been discovered. The achievements and data have been gotten in the following aspects: constitution and features of ore-forming beds and sandstone bodies, uranium existence forms and mineralogical and chemical compositions of the ores, associated elements and their economic values for comprehensive mining. The study of metallogenetic features and mechanism of the uranium deposit suggested that pre-enrichment of uranium during the depositional-diagenetic stage provide a good basis for uranium mineralization, and hereafter interlayered oxidation as well as oil-gas reduction processes played a decisive role to uranium mineralization. U-Pb isotopic studies indicate that the ores have two isochron ages of 53±3 Ma and 7.0±0 Ma, corresponding to the periods of arid and semiarid paleo-climates which are favorable to interlayered oxidation development and uranium mineralization. It is concluded that the Qianjiadian sandstone-type uranium deposit is genetically related to interlayered oxidation and secondary reduction of oil-gas. The metallogenic model of Qianjiadian uranium deposit was set up. (authors)

  7. Pb-Pb and U-Pb zircon ages of archean syntetocnic granites of the Carajas metallogenic province, northern Brazil

    International Nuclear Information System (INIS)

    Barros, Carlos Eduardo de Mesquita; Sardinha, Alex Souza; Barbosa, Jaime dos Passos de Oliveira; Krimski, Robert; Macambira, Moacir Jose Buenano

    2001-01-01

    The Carajas Metallogenic Province is located in the southeastern Amazonian Craton. It has been divided in two domains, the southernmost comprises the Rio Maria region and the northernmost corresponds to Caraj region (Souza et al. 1996). The former domain is made up of Archean greenstone sequences (2,97 Ga), TTG (2,9 Ga) and calc-alkaline granitoids (2,87 Ga) (Macambira and Lafon 1995, Leite et al. 1999, Althoff et al. 2000). The Carajas block is constituted of minor mafic granulites (3,00 Ga) and quartzofeldspathic gneisses (2,81 Ga), metavolcanosedimentary sequences (2,76 Ga) and granites (2,76 to 2,56 Ga) (Machado et al. 1991; Huhn et al. 1999, Pidgeon et al. 2000). Widespread anorogenic A-type granites are found in both areas (Docegeo 1988; Dall'Agnol et al. 1994). In the last two decades several authors (Lindenamyer et al. 1994, Barros and Barbey 1998, Huhn et al. 1999 and others) have emphasized the role of the Archean granite magmatism in the tectonicthermal evolution in the Carajas Province. In this paper we discuss the tectonic significance of the Pb- Pb and U-Pb ages obtained in some granitoids from the Carajas region. The Estrela Granite Complex and the granitoids located to the north of Parauapebas were dated by Pb- Pb evaporation zircon method (cf. Kober 1987). Data are presented considering 2σ ∼ . The Pb corrections have been done in the basis of the evolution model of Pb in double stage (cf. Stacey and Kramers 1975). U-Pb zircon method (cf. Krogh 1973, Stacey and Kramers 1975, Parrish 1987, Ludwuig 1999), recently put on routine in the Para-Iso laboratories, was employed to date the granite from the Serra do Rabo area. Analyses were carried on the Finnigan Mat 262 spectrometer (au)

  8. Characterization of a Cytokinin Response Factor in Arabidopsis thaliana

    OpenAIRE

    Ketelsen, Bernd

    2012-01-01

    The papers of this thesis are not available in Munin: 1. Bernd Ketelsen, Rainer Schwacke, Kirsten Krause and Karsten Fischer: 'Transcriptional activation by Cytokinin Response Factor 5 is governed by an acidic Cterminus containing two conserved domains' (manuscript) 2. Bernd Ketelsen, Stian Olsen, Kirsten Krause and Karsten Fischer: 'Cytokinin responsive factor 5 (CRF5) is involved in root development, hormonal crosstalk and sugar metabolism in Arabidopsis thaliana' (manuscript) 3. Bernd K...

  9. Characterization of neuritin as a novel angiogenic factor

    Energy Technology Data Exchange (ETDEWEB)

    Han, Dingding; Qin, Bo; Liu, Guoqing; Liu, Tingting; Ji, Guoqing; Wu, Yanhua [State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, Shanghai 200433 (China); Yu, Long, E-mail: longyu@fudan.edu.cn [State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, Shanghai 200433 (China)

    2011-12-02

    Highlights: Black-Right-Pointing-Pointer Neuritin protein has no effect on the endothelial cell proliferation and adhesion. Black-Right-Pointing-Pointer Neuritin protein increases endothelial cell migration. >Neuritin does not increase tumor cell proliferation in vitro. Black-Right-Pointing-Pointer Overexpression of neuritin induces tumor angiogenesis. >Overexpression of neuritin inhibits tumorigenesis. -- Abstract: Neuritin (NRN1), a neurotrophic factor, plays an important role in neurite growth and neuronal survival. In this study, we identify a new function of neuritin as a novel angiogenic factor in vitro and in vivo. Recombinant neuritin protein had no effect on the proliferation and adhesion of human umbilical vein endothelial cells (HUVEC), but it dose-dependently increased endothelial cell migration. Furthermore, overexpression of neuritin significantly promoted tumor angiogenesis, and surprisingly, it inhibited tumor growth in a xenograft tumor model. Thus, our results indicate that neuritin may act as an important angiogenic factor and serve as a potential target for cancer therapy.

  10. Molecular characterization of transforming growth factor-beta3

    NARCIS (Netherlands)

    Dijke, ten P.

    1991-01-01

    Normal tissue homeostasis is controlled by a critical balance of positive and negative modulators. Chapter 2 gives an overview of the molecular aspects of growth control, in particular the role of growth factors and oncogene and anti-oncogene products. Uncontrolled growth of cancer cells

  11. Anaerobic granular sludge : characterization, and factors affecting its functioning

    NARCIS (Netherlands)

    Alphenaar, P.A.

    1994-01-01

    Many UASB reactors are designed in such a fashion that the presence of granular sludge is necessary for a proper purification process. For achieving an optimum wastewater purification with such reactors, knowledge of the factors that determine the growth, retention and disintegration of

  12. Modern concepts for basic radiobiological factors characterizing tumor tissue radiosensitivity

    International Nuclear Information System (INIS)

    Gocheva, L.; Sergieva, K.

    2002-01-01

    Traditionally radiotherapy is prescribed at doses consistent with the expected therapeutic response and tolerance of tumor and normal tissues without consideration to individual differences in radiosensitivity. However, the basic radiobiological knowledge and clinical experience along this line point to significant variations in the observed therapeutic results. It has been established that cells and tissues under experimental and clinical conditions manifest a wide spectrum of individual radiosensitivity. The aim of this survey is to outline the current concepts for the basic radiobiological factors influencing tumor radiosensitivity. A thorough discussion is done of the essence, mechanisms of action, methods of determination and measurement, and effect on the prognosis in patients with malignant diseases of a number of radiobiological factors, such as: tumor-cell proliferation, apoptosis, tumor hypoxia and neovascularization. Although the knowledge of the mechanisms of radiosensitivity is constantly expanding, its clinical implementation is still rather limited. The true role of radiosensitivity in predicting the therapeutic response should be more accurately defined. (authors)

  13. Which Factors Characterize Sustainable Behavior of Defense Forces?

    DEFF Research Database (Denmark)

    Atlason, Reynir Smari; Gerstlberger, Wolfgang

    2017-01-01

    the behavior of 81 military forces with regard to Corporate Social Responsibility (CSR) and sustainability reporting. Furthermore, the characteristics of militaries with a CSR/sustainability strategy are identified. By using hierarchical cluster analysis and exploratory factor analysis, we locate......Defense agencies are increasingly identifying the benefits of environmentally benign solutions on multiple types of operations. Energy use, material use, and environmental damage are factors that influence military operations in both the short and the long term. In this paper, we examine...... militaries have a CSR or sustainability strategy and what the perceived benefits of such strategies are. Furthermore, we have identified militaries that are likely to implement such strategies in the future. Our findings indicate that countries with a strong regulatory environment, high Gross Domestic...

  14. Big Hitters: Important Factors Characterizing Team Effectiveness in Professional Cricket

    Directory of Open Access Journals (Sweden)

    Leonie V. Webster

    2017-07-01

    Full Text Available While organizational psychology attests to the multidimensional nature of team effectiveness, insight regarding the most important factors contributing to the effectiveness of sports teams, especially elite teams, is lacking. An abductive method of qualitative enquiry was adopted to capture participants' construal of team effectiveness, drawing on the extant literature in both sport and organizational psychology. Semi-structured interviews were conducted with 21 players, coaches, and psychologists involved in elite cricket, with resultant data analyzed inductively initially, before being reanalyzed deductively. Although, the narratives endorsed the value of many of the deductively derived factors, other constructs more prominent in organizational psychology (e.g., trust and intra-group conflict appeared to be more important than traditional sport psychology group factors. The results revealed six broad themes; culture and environment, values, communication, understanding, leadership, and unique individuals, with some gender differences apparent throughout. Based on our elite sample's construal of team effectiveness, we propose a new model representing a practical, parsimonious, and novel conceptualization of the most important attributes of team effectiveness in cricket, with conceivable transferability to other team sports.

  15. Ore-forming environment identification and uranium metallogenic features in Manite depression

    International Nuclear Information System (INIS)

    Liu Zhengyi; Liu Wusheng; Jia Licheng; Shi Qingping; Peng Cong; Chen Hua

    2014-01-01

    By using limonitization, the important indicator of uranium ore-forming identification as the product of acid water transportation from the reducing environment to the oxidation environment, based on the specimen examination naked eye, microscopic identification, electron microscopy, electron microprobe were used to study the major elements, trace elements, organic carbon, total sulfur, uranium valence and uranium, thorium isotope "2"3"4U/"2"3"8U, "2"3"0Th/"2"3"2Th ratio, hydrocarbons especially clay mineral species and the total amount, V/(V + Ni) water stratification and bottom hydrodynamic environment and other factors, which can indicate the microphase environment of all types of rocks in Manite depression, and uranium ore-forming factors were identified and evaluation are discussed and some suggestions were submitted. (authors)

  16. Intermediate sulfidation type base metal mineralization at Aliabad-Khanchy, Tarom-Hashtjin metallogenic belt, NW Iran

    Science.gov (United States)

    Kouhestani, Hossein; Mokhtari, Mir Ali Asghar; Chang, Zhaoshan; Johnson, Craig A.

    2018-01-01

    The Aliabad-Khanchy epithermal base metal deposit is located in the Tarom-Hashtjin metallogenic belt (THMB) of northwest Iran. The mineralization occurs as Cu-bearing brecciated quartz veins hosted by Eocene volcanic and volcaniclastic rocks of the Karaj Formation. Ore formation can be divided into five stages, with most ore minerals, such as pyrite and chalcopyrite being formed in the early stages. The main wall-rock alteration is silicification, and chlorite, argillic and propylitic alteration. Microthermometric measurements of fluid inclusion assemblages show that the ore-forming fluids have eutectic temperatures between −30 and −52 °C, trapping temperatures of 150–290 °C, and salinities of 6.6–12.4 wt% NaCl equiv. These data demonstrate that the ore-forming fluids were medium- to high-temperature, medium- to low-salinity, and low-density H2O–NaCl–CaCl2 fluids. Calculated δ18O values indicate that ore-forming hydrothermal fluids had δ18Owater ranging from +3.6‰ to +0.8‰, confirming that the ore–fluid system evolved from dominantly magmatic to dominantly meteoric. The calculated 34SH2S values range from −8.1‰ to −5.0‰, consistent with derivation of the sulfur from either magma or possibly from local volcanic wall-rock. Combined, the fluid inclusion and stable isotope data indicate that the Aliabad-Khanchy deposit formed from magmatic-hydrothermal fluids. After rising to a depth of between 790 and 500 m, the fluid boiled and subsequent hydraulic fracturing may have led to inflow and/or mixing of early magmatic fluids with circulating groundwater causing deposition of base metals due to dilution and/or cooling. The Aliabad-Khanchy deposit is interpreted as an intermediate-sulfidation style of epithermal mineralization. Our data suggest that the mineralization at Aliabad-Khanchy and other epithermal deposits of the THMB formed by hydrothermal activity related to shallow late Eocene magmatism. The altered Eocene volcanic and

  17. Metallogenic geologic conditions and prospecting direction of sandstone type uranium mineralizations in Yili basin of Xinjiang

    International Nuclear Information System (INIS)

    Chen Daisheng; Wang Ruiying; Li Shengxiang; Zhang Kefang

    1994-09-01

    Yili basin is a Mesozoic down-warped basin superimposed on the late Paleozoic volcanic taphrogenic basin. Uranium mineralizations are hosted in the Middle-Lower Jurassic coal-bearing series. The depositions environment in the basin is turbulent in the east and relatively stable in the west. It is characterized by coarse-grained sequence with thin thickness in the eastern part and fine-grained with thick thickness in the western part. On the analytical basis of sedimentary facies indices, it is the first time to present a sedimentary model of 'alluvial fan-braided stream-(narrow) lakeshore delta-lacustrine facies and marsh facies' for the coal-bearing series. The authors have summarized the basic geologic features of U-mineralizations in the interlayer oxidation zone, analyzed the difference and cause of U-mineralizations between the south and north, as well as the east and west. The genetic mechanism of U-mineralizations in the basin is discussed. Finally, seven items of geologic prerequisites for the formation of in-situ leachable sandstone type uranium deposits have been suggested and the potential of sandstone type U-mineralizations in the basin has been evaluated. Four promising target areas are selected

  18. Reviews on the metallogenic and geological features of sandstone-type uranium deposits in Japan

    International Nuclear Information System (INIS)

    Pei Chengkai; Huang Xianfang; Zhang Baoju

    2006-01-01

    Regional geologic settings of sandstone-type uranium deposits in Japan are firstly analyzed. The regional tectonic evolution characteristics of 'Green tuff region' and 'Non green tuff region' and their relationship with uranium mineralization are elaborated in depth. Based on those mentioned above, the uranium sources of sandstone-type uranium deposits in Japan are discussed deeply and the most favorable uranium sources are considered to come from the basement and the surrounding granites. Their intrusive epochs range from Later Cretaceous to Palaeogene (about 60 to 70 Ma ago). The characteristics of ore-bearing host rocks, matter compositions of the deposits, ore formation enrichment factors, the hydrogeologic conditions and so on are described by taking Ningyo-Toge and Tono deposits as examples. Finally, the prospecting measures for the palaeo-channel sandstone-type uranium deposits (basal type) are reviewed. (authors)

  19. Characterization of the Far Transcription Factor Family in Aspergillus flavus.

    Science.gov (United States)

    Luo, Xingyu; Affeldt, Katharyn J; Keller, Nancy P

    2016-10-13

    Metabolism of fatty acids is a critical requirement for the pathogenesis of oil seed pathogens including the fungus Aspergillus flavus Previous studies have correlated decreased ability to grow on fatty acids with reduced virulence of this fungus on host seed. Two fatty acid metabolism regulatory transcription factors, FarA and FarB, have been described in other filamentous fungi. Unexpectedly, we find A. flavus possesses three Far homologs, FarA, FarB, and FarC, with FarA and FarC showing a greater protein similarity to each other than FarB. farA and farB are located in regions of colinearity in all Aspergillus spp. sequenced to date, whereas farC is limited to a subset of species where it is inserted in an otherwise colinear region in Aspergillus genomes. Deletion and overexpression (OE) of farA and farB, but not farC, yielded mutants with aberrant growth patterns on specific fatty acids as well as altered expression of genes involved in fatty acid metabolism. Marked differences included significant growth defects of both ∆farA and ∆farB on medium-chain fatty acids and decreased growth of OE::farA on unsaturated fatty acids. Loss of farA diminished expression of mitochondrial β-oxidation genes whereas OE::farA inhibited expression of genes involved in unsaturated fatty acid catabolism. FarA also positively regulated the desaturase genes required to generate polyunsaturated fatty acids. Aflatoxin production on toxin-inducing media was significantly decreased in the ∆farB mutant and increased in the OE::farB mutant, with gene expression data supporting a role for FarB in tying β-oxidation processes with aflatoxin accumulation. Copyright © 2016 Luo et al.

  20. Characterization of the Far Transcription Factor Family in Aspergillus flavus

    Directory of Open Access Journals (Sweden)

    Xingyu Luo

    2016-10-01

    Full Text Available Metabolism of fatty acids is a critical requirement for the pathogenesis of oil seed pathogens including the fungus Aspergillus flavus. Previous studies have correlated decreased ability to grow on fatty acids with reduced virulence of this fungus on host seed. Two fatty acid metabolism regulatory transcription factors, FarA and FarB, have been described in other filamentous fungi. Unexpectedly, we find A. flavus possesses three Far homologs, FarA, FarB, and FarC, with FarA and FarC showing a greater protein similarity to each other than FarB. farA and farB are located in regions of colinearity in all Aspergillus spp. sequenced to date, whereas farC is limited to a subset of species where it is inserted in an otherwise colinear region in Aspergillus genomes. Deletion and overexpression (OE of farA and farB, but not farC, yielded mutants with aberrant growth patterns on specific fatty acids as well as altered expression of genes involved in fatty acid metabolism. Marked differences included significant growth defects of both ∆farA and ∆farB on medium-chain fatty acids and decreased growth of OE::farA on unsaturated fatty acids. Loss of farA diminished expression of mitochondrial β-oxidation genes whereas OE::farA inhibited expression of genes involved in unsaturated fatty acid catabolism. FarA also positively regulated the desaturase genes required to generate polyunsaturated fatty acids. Aflatoxin production on toxin-inducing media was significantly decreased in the ∆farB mutant and increased in the OE::farB mutant, with gene expression data supporting a role for FarB in tying β-oxidation processes with aflatoxin accumulation.

  1. Geochronology and petrogenesis of the Qibaoshan Cu-polymetallic deposit, northeastern Hunan Province: Implications for the metal source and metallogenic evolution of the intracontinental Qinhang Cu-polymetallic belt, South China

    Science.gov (United States)

    Yuan, Shunda; Mao, Jingwen; Zhao, Panlao; Yuan, Yabin

    2018-03-01

    The recently recognized Qinhang metallogenic belt (QHMB) is an economically important intracontinental Mesozoic porphyry-skarn Cu-polymetallic metallogenic belt in South China. However, the origin of the ore-bearing magma and the major factors controlling the different metal assemblages in the QHMB are still unclear. The Qibaoshan deposit is a large Cu-Au-Pb-Zn-Ag-Fe deposit located at the juncture between the northern and central parts of the QHMB. In this study, new zircon U-Pb ages, Hf-O isotopic data, molybdenite Re-Os ages, and whole-rock geochemical data are combined to constrain the timing of the mineralization and the origin and petrogenesis of the ore-bearing porphyry in the Qibaoshan deposit. The ages obtained from both zircon U-Pb and molybdenite Re-Os dating fall in the Late Jurassic (between 152.7 and 148.3 Ma), revealing that this deposit is significantly younger than previously estimated (227-184 Ma). The Qibaoshan ore-bearing quartz porphyry shows variable negative zircon εHf(t) values (-14.8 to -5.5), high δ18O values (8.4 to 10.8‰), and high Mg# values (69.1 to 73.0), indicating that it formed via the partial melting of ancient crust triggered by the injection of mantle-derived magma. Zircon Hf-O isotopic modeling of the mixing of two extreme endmembers indicates that the magmatic source comprised 70-80% reworked ancient crustal components and 20-30% depleted mantle components. Based on comparisons with other ore-bearing porphyries in the QHMB, a magmatic source dominated by crust-derived material and relatively low oxygen fugacities (ΔFMQ -1.8 to ΔFMQ +0.8) was responsible for the high (Pb + Zn)/Cu ratio in the Qibaoshan deposit, and the Pb, Zn and Ag were mainly derived from the reworked ancient crust. Although four analyses of inherited Neoproterozoic zircons ( 800 Ma) have variable positive εHf(t) values (0.72 to 11.21), indicating that Neoproterozoic juvenile crust was involved in the formation of the Qibaoshan ore-bearing quartz

  2. Regional evaluation and primary geological structural and metallogenical research of great Kavir basin as view of possibility formation of sedimentary-surficial Uranium mineralization

    International Nuclear Information System (INIS)

    Kamali Sadr, S.

    2006-01-01

    Great Kavir basin is the largest inner basin in Iran that extended about 90000 km 2. This basin is situated in the centre of lran , to the south from Alborz mountain range and elongated in the sub- latitudinal trend and its construction is asymmetric. The basin cover consists generally of complicated sequence of continental - marine Oligocene - Miocene molasses. According to drainage systems - conditions, molassoid cycles, alluvial, alluvial - deltaic and lacustrine sediments, climate, morphological conditions and metallogenic and structural features, Great Kavir depression generally is favorable for exigence and surficial uranium deposits (vally - fill, flood plain, deltaic and playa). Uranium occurrences that are Known in the southern and north eastern part of the margent Great Kavir basin, are Arosan, Irekan and Mohammad Abad. Similar geological - structural conditions for uranium mineralization is possible in the margent of Great Kavir basin

  3. Controlling factors of uranium mineralization and prospect prediction in Qimantage area

    International Nuclear Information System (INIS)

    Yao Chunling; Zhu Pengfei; Cai Yuqi; Zhang Wenming; Zhao Yong'an; Song Jiye; Zhang Xiaojin

    2011-01-01

    Based on the analysis of regional geology in Qimantage area, the condition for uranium mineralization is summarized in regional geology setting, volcanic, granite and faults. This study shows that this area has favorable prospect for uranium mineralization. The metallogenic model is built up according to the controlling factors over uranium mineralization. Under this model, six potential areas are predicted in MRAS software with mineralization factors of synthetically geological information method. (authors)

  4. Molecular and functional characterization of pigeon (Columba livia) tumor necrosis factor receptor-associated factor 3.

    Science.gov (United States)

    Zhou, Yingying; Kang, Xilong; Xiong, Dan; Zhu, Shanshan; Zheng, Huijuan; Xu, Ying; Guo, Yaxin; Pan, Zhiming; Jiao, Xinan

    2017-04-01

    Tumor necrosis factor receptor-associated factor 3 (TRAF3) plays a key antiviral role by promoting type I interferon production. We cloned the pigeon TRAF3 gene (PiTRAF3) according to its predicted mRNA sequence to investigate its function. The 1704-bp full-length open reading frame encodes a 567-amino acid protein. One Ring finger, two TRAF-type Zinc fingers, one Coiled coil, and one MATH domain were inferred. RT-PCR showed that PiTRAF3 was expressed in all tissues, with relatively weak expression in the heart and liver. In HEK293T cells, over-expression of wild-type, △Ring, △Zinc finger, and △Coiled coil PiTRAF3, but not a △MATH form, significantly increased IFN-β promoter activity. Zinc finger and Coiled coil domains were essential for NF-κB activation. In chicken HD11 cells, PiTRAF3 increased IFN-β promoter activity and four domains were all contributing. R848 stimulation of pigeon peripheral blood mononuclear cells and splenocytes significantly increased expression of PiTRAF3 and the inflammatory cytokine genes CCL5, IL-8, and IL-10. These data demonstrate TRAF3's innate immune function and improve understanding of its involvement in poultry antiviral defense. Copyright © 2016 Elsevier Ltd. All rights reserved.

  5. Molecular characterization of Candida in the oral cavity and factors involved in biofilm formation and virulence

    NARCIS (Netherlands)

    Kraneveld, E.A.

    2014-01-01

    The research described in this thesis addresses current issues related to oral Candida infections. Interactions of Candida with the oral microbiome were characterized and factors involved in biofilm formation and virulence were studied. All in all, the work described in this thesis contributes

  6. Partial characterization of a putative new growth factor present in pathological human vitreous.

    Science.gov (United States)

    Pombo, C; Bokser, L; Casabiell, X; Zugaza, J; Capeans, M; Salorio, M; Casanueva, F

    1996-03-01

    Several growth factors have been implicated in the development of proliferative eye diseases, and some of those are present in human vitreous (HV). The effects of HV on cellular responses which modulate proliferative cell processes were studied. This study describes the partial characterization of a vitreous factor activity which does not correspond to any of the previously reported growth factors in pathological HV. Vitreous humour was obtained from medical vitrectomies, from patients with PDR and PVR. The biological activity of the vitreous factor was determined by its ability to increase cytosolic calcium concentration ([Ca2+]i), increase production of inositol phosphates, and induce cell proliferation in the cell line EGFR T17. In some experiments other cell lines, such as NIH 3T3, 3T3-L1, FRTL5, A431, PC12, Y79, and GH3, were also employed. Measurement of [Ca2+]i in cell suspensions was performed using the fluorescent Ca2+ indicator fura-2. The activity of the factor present in HV was compared with other growth factors by means of: (a) [Ca2+]i mobilization pattern, (b) sequential homologous and heterologous desensitization of receptors, (c) effects of phorbol esters on their action, and (d) inactivation after treatment with different proteolytic enzymes. The HV-induced cell proliferation and increases in [Ca2+]i concentration were characterized by a peculiar time pattern. The different approaches used ruled out its identity with PDGF, bFGF, EGF, TGF-beta, IGFs, TNF-alpha, NGF, and other compounds such as ATP, angiotensin I, and bradykinin. Vitreous factor actions are mediated by specific receptors apparently regulated by PKC. This factor is able to induce [Ca2+]i mobilization in most of the cell lines studied, indicating that its effects are not tissue specific. These results suggest the presence of a growth factor activity in pathological HV which may be due to the presence of an undescribed growth factor in the eye.

  7. Geochemistry and geochronology of ore-bearing and barren intrusions in the Luanchuan ore fields of East Qinling metallogenic belt, China: Diverse tectonic evolution and implications for mineral exploration

    Science.gov (United States)

    Xue, Fei; Wang, Gongwen; Santosh, M.; Yang, Fan; Shen, Zhiwei; Kong, Liang; Guo, Nana; Zhang, Xuhuang; Jia, Wenjuan

    2018-05-01

    The Luanchuan ore fields form part of the East Qinling metallogenic belt in central China. In this study, we compare two ore-bearing intrusions, the Shibaogou granitic pluton (SBG) and the Zhongyuku granitic pluton (ZYK), with the ore-barren Laojunshan intrusion (LJS) from the Luanchuan ore field. Geochemically, all the three intrusions are characterized by high-Si, high-K, and alkalis, together with moderate-ASI, exhibiting I-type granite features. The rocks, especially the ore-related plutons also show enrichment in LREEs. Mineral chemistry of biotite from the intrusions exhibits similar features of high Si and Mg, and low Al and Fe. Zircon grains from the ZYK intrusion yielded a U-Pb age of 149.6 ± 2.4 Ma. The zircon grains show εHf (t) values and two stage model ages (TDM2) in the range of -16.8 to -19.7 and 1998-2156 Ma respectively. The biotite composition and Hf isotopic data indicate that the magma was derived by re-melting of deep crustal material with minor input of mantle components. We evaluate the results to understand the physico-chemical conditions, petrogenesis, and tectonic setting, and their implications for mineral exploration. The ore-bearing plutons show wide ranges of temperature and oxygen fugacity, favoring Mo-W mineralization. In addition, estimates on pressure and depth of emplacement suggest that lower solidification pressure in a decompressional setting contributed to the evolution of magmatic hydrothermal deposits. Our data suggest that the ZYK has the highest potential for Mo-W mineralization. The ore-bearing plutons of ZYK and SBG were formed in a transitional tectonic setting from compression to extension, with the large-scale metallogeny triggered by slab melts at ca. 145 Ma. However, the ore-barren LJS batholith formed in an extension-related geodynamic setting at ∼115 Ma. Our study shows that different tectonic settings and consequent physico-chemical conditions dictated the ore potential of the intrusions in the Luanchuan ore

  8. Investigation on the age of mineralization in the Sungun porphyry Cu-Mo deposit, NW Iran with a regional metallogenic perspective

    Science.gov (United States)

    Simmonds, Vartan; Moazzen, Mohssen; Mathur, Ryan

    2016-04-01

    The Sungun porphyry copper deposit (PCD) is located in NW Iran, neighbouring several other PCDs and prospects in the region and the Lesser Caucasus (south Armenia). It lies on the Urumieh-Dokhtar magmatic arc (UDMA), which formed through the northeast-ward subduction of the Neo-Tethyan oceanic crust beneath the Central Iranian plate during late-Mesozoic and early-Cenozoic [1], and hosts the porphyry copper metallogenic belt of Iran. The Sungun PCD is the second largest deposit in Iran with ore reserves of about 850 Mt at 0.62 wt% Cu and 0.01 wt% Mo and probable reserves over 1Gt. The monzonitic to quartz monzonitic porphyry stock intruded the upper Cretaceous carbonates and Eocene volcano-sedimentary rocks. It produced a skarn-type mineralization at its contact zone with the carbonate rocks, as well as vast hydrothermal alteration zones and porphyry-type Cu and Mo mineralization. The zircon U-Pb age of the host porphyry stock is about 22.5±0.4 to 20.1±0.4 Ma [2]. Re-Os dating of four molybdenite separates from this PCD shows ages ranging between 22.9±0.2 to 21.7±0.2 Ma, with an average of 22.57±0.2 Ma, corresponding to the early Miocene (Aquitanian). These ages indicate that both the porphyry stock and the Cu-Mo mineralization are post-collisional events, similar to many other deposits and prospects in NW and central Iran and south Armenia, and the mineralization occurred shortly after the emplacement of the host stock, corresponding better to the ages obtained from the marginal parts of the stock. Magmatism and mineralization in Sungun coincides with the third metallogenic epoch in the Lesser Caucasus (Eocene to Miocene; [3]), though it is considerably younger than all of the dated PCDs and prospects in the south Armenia. It also postdates Cu-Mo mineralizations in the Saheb Divan (35 Ma), Qaradagh batholith (31.22±0.28 to 25.19±0.19 Ma), as well as Haft Cheshmeh PCD (28.18±0.42 to 27.05±0.37 Ma) in NW Iran, while it seems to be coeval with the Kighal

  9. Towards factor analysis exploration applied to positron emission tomography functional imaging for breast cancer characterization

    International Nuclear Information System (INIS)

    Rekik, W.; Ketata, I.; Sellami, L.; Ben slima, M.; Ben Hamida, A.; Chtourou, K.; Ruan, S.

    2011-01-01

    This paper aims to explore the factor analysis when applied to a dynamic sequence of medical images obtained using nuclear imaging modality, Positron Emission Tomography (PET). This latter modality allows obtaining information on physiological phenomena, through the examination of radiotracer evolution during time. Factor analysis of dynamic medical images sequence (FADMIS) estimates the underlying fundamental spatial distributions by factor images and the associated so-called fundamental functions (describing the signal variations) by factors. This method is based on an orthogonal analysis followed by an oblique analysis. The results of the FADMIS are physiological curves showing the evolution during time of radiotracer within homogeneous tissues distributions. This functional analysis of dynamic nuclear medical images is considered to be very efficient for cancer diagnostics. In fact, it could be applied for cancer characterization, vascularization as well as possible evaluation of response to therapy.

  10. Characterization factors for global warming in life cycle assessment based on damages to humans and ecosystems.

    Science.gov (United States)

    De Schryver, An M; Brakkee, Karin W; Goedkoop, Mark J; Huijbregts, Mark A J

    2009-03-15

    Human and ecosystem health damage due to greenhouse gas (GHG) emissions is generally poorly quantified in the life cycle assessment of products, preventing an integrated comparison of the importance of GHGs with other stressor types, such as ozone depletion and acidifying emissions. In this study, we derived new characterization factors for 63 GHGs that quantify the impact of an emission change on human and ecosystem health damage. For human health damage, the Disability Adjusted Life Years (DALYs) per unit emission related to malaria, diarrhea, malnutrition, drowning, and cardiovascular diseases were quantified. For ecosystem health damage, the Potentially Disappeared Fraction (PDF) over space and time of various species groups, including plants, butterflies, birds, and mammals, per unit emission was calculated. The influence of value choices in the modeling procedure was analyzed by defining three coherent scenarios, based on Cultural theory perspectives. It was found that the characterization factor for human health damage by carbon dioxide (CO2) ranges from 1.1 x 10(-2) to 1.8 x 10(+1) DALY per kton of emission, while the characterization factor for ecosystem damage by CO2 ranges from 5.4 x 10(-2) to 1.2 x 10(+1) disappeared fraction of species over space and time ((km2 x year)/kton), depending on the scenario chosen. The characterization factor of a GHG can change up to 4 orders of magnitude, depending on the scenario. The scenario-specific differences are mainly explained by the choice for a specific time horizon and stresses the importance of dealing with value choices in the life cycle impact assessment of GHG emissions.

  11. Loregic: A Method to Characterize the Cooperative Logic of Regulatory Factors

    Science.gov (United States)

    Wang, Daifeng; Yan, Koon-Kiu; Sisu, Cristina; Cheng, Chao; Rozowsky, Joel; Meyerson, William; Gerstein, Mark B.

    2015-01-01

    The topology of the gene-regulatory network has been extensively analyzed. Now, given the large amount of available functional genomic data, it is possible to go beyond this and systematically study regulatory circuits in terms of logic elements. To this end, we present Loregic, a computational method integrating gene expression and regulatory network data, to characterize the cooperativity of regulatory factors. Loregic uses all 16 possible two-input-one-output logic gates (e.g. AND or XOR) to describe triplets of two factors regulating a common target. We attempt to find the gate that best matches each triplet’s observed gene expression pattern across many conditions. We make Loregic available as a general-purpose tool (github.com/gersteinlab/loregic). We validate it with known yeast transcription-factor knockout experiments. Next, using human ENCODE ChIP-Seq and TCGA RNA-Seq data, we are able to demonstrate how Loregic characterizes complex circuits involving both proximally and distally regulating transcription factors (TFs) and also miRNAs. Furthermore, we show that MYC, a well-known oncogenic driving TF, can be modeled as acting independently from other TFs (e.g., using OR gates) but antagonistically with repressing miRNAs. Finally, we inter-relate Loregic’s gate logic with other aspects of regulation, such as indirect binding via protein-protein interactions, feed-forward loop motifs and global regulatory hierarchy. PMID:25884877

  12. Metal toxicity characterization factors for marine ecosystems: considering the importance of the estuary for freshwater emissions

    DEFF Research Database (Denmark)

    Dong, Yan; Rosenbaum, Ralph K.; Hauschild, Michael Zwicky

    2017-01-01

    The study develops site-dependent characterization factors (CFs) for marine ecotoxicity of metals emitted to freshwater, taking their passage of the estuary into account. To serve life cycle assessment (LCA) studies where emission location is often unknown, site-generic marine CFs were developed...... with an estuary removal process to calculate FF. BF and EF were taken from Dong et al. Environ Sci Technol 50:269–278 (2016). Site-generic marine CFs were derived from site-dependent marine CFs. Different averaging principles were tested, and the approach representing estuary discharge rate was identified...... between both methods. Accounting for estuary removal particularly influences marine ecotoxicity CFs for emission to freshwater of metals that have a strong tendency to complex-bind to particles. It indicates the importance of including estuary in the characterization modelling when dealing with those...

  13. Identification and Characterization of Soluble Factors Involved in Delayed Effects of Low Dose Radiation. Final report

    International Nuclear Information System (INIS)

    Baulch, Janet

    2013-01-01

    This is a 'glue grant' that was part of a DOE Low Dose project entitled 'Identification and Characterization of Soluble Factors Involved in Delayed Effects of Low Dose Radiation'. This collaborative program has involved Drs. David L. Springer from Pacific Northwest National Laboratory (PNNL), John H. Miller from Washington State University, Tri-cities (WSU) and William F. Morgan then from the University of Maryland, Baltimore (UMB). In July 2008, Dr. Morgan moved to PNNL and Dr. Janet E. Baulch became PI for this project at University of Maryland. In November of 2008, a one year extension with no new funds was requested to complete the proteomic analyses. The project stemmed from studies in the Morgan laboratory demonstrating that genomically unstable cells secret a soluble factor or factors into the culture medium, that cause cytogenetic aberrations and apoptosis in normal parental GM10115 cells. The purpose of this project was to identify the death inducing effect (DIE) factor or factors, estimate their relative abundance, identify the cell signaling pathways involved and finally recapitulate DIE in normal cells by exogenous manipulation of putative DIE factors in culture medium. As reported in detail in the previous progress report, analysis of culture medium from the parental cell line, and stable and unstable clones demonstrated inconsistent proteomic profiles as relate to candidate DIE factors. While the proposed proteomic analyses did not provide information that would allow DIE factors to be identified, the analyses provided another important set of observations. Proteomic analysis suggested that proteins associated with the cellular response to oxidative stress and mitochondrial function were elevated in the medium from unstable clones in a manner consistent with mitochondrial dysfunction. These findings correlate with previous studies of these clones that demonstrated functional differences between the mitochondria of stable and unstable clones. These

  14. Identification and Characterization of Soluble Factors Involved in Delayed Effects of Low Dose Radiation. Final report

    Energy Technology Data Exchange (ETDEWEB)

    Baulch, Janet

    2013-09-11

    This is a 'glue grant' that was part of a DOE Low Dose project entitled 'Identification and Characterization of Soluble Factors Involved in Delayed Effects of Low Dose Radiation'. This collaborative program has involved Drs. David L. Springer from Pacific Northwest National Laboratory (PNNL), John H. Miller from Washington State University, Tri-cities (WSU) and William F. Morgan then from the University of Maryland, Baltimore (UMB). In July 2008, Dr. Morgan moved to PNNL and Dr. Janet E. Baulch became PI for this project at University of Maryland. In November of 2008, a one year extension with no new funds was requested to complete the proteomic analyses. The project stemmed from studies in the Morgan laboratory demonstrating that genomically unstable cells secret a soluble factor or factors into the culture medium, that cause cytogenetic aberrations and apoptosis in normal parental GM10115 cells. The purpose of this project was to identify the death inducing effect (DIE) factor or factors, estimate their relative abundance, identify the cell signaling pathways involved and finally recapitulate DIE in normal cells by exogenous manipulation of putative DIE factors in culture medium. As reported in detail in the previous progress report, analysis of culture medium from the parental cell line, and stable and unstable clones demonstrated inconsistent proteomic profiles as relate to candidate DIE factors. While the proposed proteomic analyses did not provide information that would allow DIE factors to be identified, the analyses provided another important set of observations. Proteomic analysis suggested that proteins associated with the cellular response to oxidative stress and mitochondrial function were elevated in the medium from unstable clones in a manner consistent with mitochondrial dysfunction. These findings correlate with previous studies of these clones that demonstrated functional differences between the mitochondria of stable and

  15. Purification and partial characterization of an early pregnancy factor-induced suppressor factor (EPF-S1).

    Science.gov (United States)

    Rolfe, B A; Athanasas-Platsis, S; Hoskin, M J; Morton, H; Cavanagh, A C

    1995-06-01

    The immunomodulatory properties of early pregnancy factor (EPF) are mediated through induction of at least two lymphokines, designated EPF-S1 and EPF-S2 (previously estimated M(r) 15,000 and 55,000 respectively). The activity of the former is MHC-restricted while the latter is restricted to a locus (or loci) outside the MHC. The present study established further criteria by which EPF-S1 and EPF-S2 might be distinguished from each other and compared with other suppressor factors. In addition, techniques have been developed to purify EPF-S1 to homogeneity. Congenic mouse strains were used to map the genetic restriction of EPF-S2 in the rosette inhibition test and high performance gel permeation chromatography was used to demonstrate that EPF-S1 induces EPF-S2 but not vice versa. Further studies then focused on isolation of this first component of the cascade, EPF-S1, from immune ascites (from growth in athymic mice of the anti-EPF-S1 producing rat-mouse hybridoma R2T gamma, in which EPF-S1 is complexed to antibody). Techniques used were acidification followed by application to Sep-pak C18 cartridges, high performance cation-exchange chromatography and two reversed-phased HPLC steps on a C3 column. Purified material was analyzed by SDS-PAGE and Edman degradation. Approximately 10 micrograms EPF-S1 were isolated fom 60 ml ascitic fluid. Homogeneity of the purified material was demonstrated by SDS-PAGE, where it ran as a single band of approximate M(r) 12,000 coincident with biological activity. Attempts at Edman degradation indicate that the molecule is N-blocked. Definitive primary characterization of EPF-S1 must await the preparation and isolation of proteolytic fragments of the molecule, but the present studies establish conditions which make such structural analysis possible.

  16. Low back pain characterized by muscle resistance and occupational factors associated with nursing

    Directory of Open Access Journals (Sweden)

    Rafael de Souza Petersen

    2014-06-01

    Full Text Available OBJECTIVE: to identify the occupational factors associated with low back pain using a surveillance tool and to characterize the low back pain by the resistance of the extensor muscles of the vertebral column among nursing professionals at an Intensive Care Unit.METHODS: Cross-sectional study. The workers answered a questionnaire about occupational factors and participated in a resistance test of the extensor muscles of the vertebral column. Associations were established through Student's T-test or Mann-Whitney's U-test and correlations using Pearson's test.RESULTS: Out of 48 participants, 32 (67% suffered from low pain. For the resistance test, the subjects suffering from low back pain endured less time in comparison with asymptomatic subjects, but without significant differences (p=0.147. The duration of the pain episode showed a significant negative correlation (p=0.016 with the results of the resistance test though. The main factors identified as causes of low back pain were biomechanical and postural elements, conditions of the muscle structure and physical and organizational conditions.CONCLUSIONS: the main occupational factors associated with the low back pain were the posture and the characteristics of the physical and organizational conditions. In addition, the extensor muscles of the column showed a trend towards lesser resistance for workers in pain. This evidence is important when considering prevention and treatment strategies.

  17. Using apatite to discriminate synchronous ore-associated and barren granitoid rocks: A case study from the Edong metallogenic district, South China

    Science.gov (United States)

    Duan, Deng-Fei; Jiang, Shao-Yong

    2018-06-01

    In order to find criteria to discriminate the synchronous ore-associated and barren granitoid rocks, we have determined apatite chemistry associated with ore-associated (Cu-Au) and barren granitoid rocks in the Edong district of the Middle and Lower Yangtze River metallogenic belt, South China. Both rock types give zircon U-Pb ages between 135.0 and 138.7 Ma. Apatite has a higher volatile and Li content (Cl: 0.19-0.57 wt%, average 0.35 wt%, SO3: 0.08-0.71 wt%, average 0.32 wt%, Li: 0.49-7.99 ppm, average 3.23 ppm) in ore-associated rocks than those in barren rocks (Cl: 0.09-0.31 wt%, average 0.16 wt%, SO3: 0.06-0.28 wt%, average 0.16 wt%, Li: 0.15-0.89 ppm, average 0.36 ppm). Apatite (La/Yb)N ratios and Eu/Eu* values are relatively high and show wider variation in ore-associated rocks than those in barren rocks. Apatite (La/Sm)N and (Yb/Sm)N show positive correlation in ore-associated rocks but negative in barren rocks. The higher volatile content occurs in ore-associated magma, favoring Cu-Au transportation and deposition. Furthermore, amphibole fractional crystallization in ore-associated magma further enriched the ore elements in the residual melt. Barren rocks may have undergone fluid exsolution before emplacement, which makes it barren in Cl, S and ore elements (Cu, S). These signatures emphases the significance of volatile and magma evolution in mineralization and indicate that analyses of magmatic apatite can serve to distinguish ore-associated from barren intrusions.

  18. Zircon U-Pb dating of Maherabad porphyry copper-gold prospect area: evidence for a late Eocene porphyry-related metallogenic epoch in east of Iran

    Directory of Open Access Journals (Sweden)

    Azadeh Malekzadeh Shafaroudi

    2011-04-01

    Full Text Available Eastern Iran has great potential for porphyry copper deposits, as a result of its past subduction zone tectonic setting that lead to extensive alkaline to calc-alkaline magmatic activity in Tertiary time. Maherabad is the first porphyry Cu-Au prospecting area which is discovered in eastern Iran. This is related to a succession o f monzonitic to dioritic porphyries stocks that were emplaced within volcanic rocks. Monzonitic porphyries have basic role in mineralization. Hydrothermal alteration zones are well developed including potassic, sericitic-potassic, quartz-sericite-carbonate-pyrite, quartz-carbonate-pyrite, silicified-propylitic, propylitic, carbonate and silicified zones. Mineralization occurs as Disseminated, stockwork and hydrothermal breccia. Based on early stage of exploration, Cu is between 179- 6830 ppm (ave. 3200 ppm and Au is up to 1000 ppb (ave. 570 ppb. This prospect is gold- rich porphyry copper deposit. Laser-ablation U-Pb dating of two samples from ore-related intrusive rocks indicate that these two monzonitic porphyries crystallized at 39.0 ± 0.8 Ma to 38.2 ± 0.8 Ma, within a short time span of less than ca. 1 Ma during the middle Eocene. This provides the first precise ages for metallogenic episode of porphyry-type mineralization. Also, the initial 87Sr/86Sr and (143Nd/144Ndi was recalculated to an age of 39 Ma. Initial 87Sr/86Sr ratios for monzonitic rocks are 0.7047-0.7048. The (143Nd/144Ndi isotope composition are 0.512694-0.512713. Initial ε Nd isotope values 1.45-1.81. Based on isotopic data the magma had originated beyond the continental crust. The study will be used for tectonic-magmatic setting and evolution of eastern Iran. Keywords: Lut block, Middle Eocene, Zircon, Geochronology, Laser ablation ICP-MS,

  19. Metallogenic condition and regularity of inter layered oxidation zone-type sandstone uranium deposit in southwestern part of Turpan-Hami basin, Northwestern China

    International Nuclear Information System (INIS)

    Xiang Weidong; Chen Zhaobo; Chen Zuyi; Yin Jinshuang

    2001-01-01

    Regional geological surveying and drilling evaluation in recent years show that there are very large potential resources of sandstone-type uranium deposits in the southwestern part of Turpan-Hami basin. According to the characteristics of tectonic evolution and sedimentary cover of the basin, the evolution stages and types of the basin are divided, and the favorable development stages for the ore-bearing formation and the formation of uranium deposits in the evolution process are identified. The metallogenic conditions of uranium deposits are deeply discussed from four aspects: basic tectonics, paleoclimate evolution, hydrogeology and uranium source of the region. All these have laid an important foundation for accurate prediction and evaluation of uranium resources in this region. The research indicates that the uranium metallogeny is a process of long-term, multi-stage and pulsation. The authors try to ascertain the role of organic matter in concentrating uranium. The organic matter is of humic type in sandstone host-rock in the studied area, whose original mother material mainly belongs to terrestrial high plant. The maturity of the organic matter is very low, being in low-grade stage of thermal evolution. Correlation analysis and separation experiments show that uranium concentration is closely related with the organic matter, and the organic matter in uranium ore is mainly in the form of humic acid adsorption and humate. For this reason the total organic carbon content is often increased in the geochemical redox zone in epigenetic sandstone-type uranium deposits. It is suggested that the north of China is of great potential for sandstone-type uranium resources

  20. Variation of molybdenum isotopes in molybdenite from porphyry and vein Mo deposits in the Gangdese metallogenic belt, Tibetan plateau and its implications

    Science.gov (United States)

    Wang, Yong; Zhou, Lian; Gao, Shan; Li, Jian-Wei; Hu, Zhi-Fang; Yang, Lu; Hu, Zhao-Chu

    2016-02-01

    We present Mo isotopic ratios of molybdenite from five porphyry molybdenum deposits (Chagele, Sharang, Jiru, Qulong, and Zhuonuo) and one quartz-molybdenite vein-type deposit (Jigongcun) along the Gangdese metallogenic belt in the Tibetan Plateau. These deposits represent a sequence of consecutive events of the India-Asia collision at different periods. Additional molybdenite samples from the Henderson Mo deposit (USA), the oceanic subduction-related El Teniente (Chile), and Bingham (USA) porphyry Cu-(Mo) deposits were analyzed for better understanding the controls on the Mo isotope systematics of molybdenite. The results show that molybdenite from Sharang, Jiru, Qulong, and Zhuonuo deposits have similar δ97Mo (˜0 ‰), in agreement with the values of the Henderson Mo deposit (-0.10 ‰). In contrast, samples from the Changle and Jigongcun deposit have δ97Mo of 0.85 ‰ to 0.88 ‰ and -0.48 %, respectively. Molybdenite from the El Teniente and Bingham deposits yields intermediate δ97Mo of 0.27 and 0.46 ‰, respectively. The Mo isotopes, combined with Nd isotope data of the ore-bearing porphyries, indicate that source of the ore-related magmas has fundamental effects on the Mo isotopic compositions of molybdenite. Our study indicates that molybdenite related to crustal-, and mantle-derived magmas has positive or negative δ97Mo values, respectively, whereas molybdenite from porphyries formed by crust-mantle mixing has δ97Mo close to 0 ‰. It is concluded that the Mo isotope composition in the porphyry system is a huge source signature, without relation to the tectonic setting under which the porphyry deposits formed.

  1. Functional Characterization of Tea (Camellia sinensis MYB4a Transcription Factor Using an Integrative Approach

    Directory of Open Access Journals (Sweden)

    Mingzhuo Li

    2017-06-01

    Full Text Available Green tea (Camellia sinensis, Cs abundantly produces a diverse array of phenylpropanoid compounds benefiting human health. To date, the regulation of the phenylpropanoid biosynthesis in tea remains to be investigated. Here, we report a cDNA isolated from leaf tissues, which encodes a R2R3-MYB transcription factor. Amino acid sequence alignment and phylogenetic analysis indicate that it is a member of the MYB4-subgroup and named as CsMYB4a. Transcriptional and metabolic analyses show that the expression profile of CsMYB4a is negatively correlated to the accumulation of six flavan-3-ols and other phenolic acids. GFP fusion analysis shows CsMYB4a’s localization in the nucleus. Promoters of five tea phenylpropanoid pathway genes are isolated and characterized to contain four types of AC-elements, which are targets of MYB4 members. Interaction of CsMYB4a and five promoters shows that CsMYB4a decreases all five promoters’ activity. To further characterize its function, CsMYB4a is overexpressed in tobacco plants. The resulting transgenic plants show dwarf, shrinking and yellowish leaf, and early senescence phenotypes. A further genome-wide transcriptomic analysis reveals that the expression levels of 20 tobacco genes involved in the shikimate and the phenylpropanoid pathways are significantly downregulated in transgenic tobacco plants. UPLC-MS and HPLC based metabolic profiling reveals significant reduction of total lignin content, rutin, chlorogenic acid, and phenylalanine in CsMYB4a transgenic tobacco plants. Promoter sequence analysis of the 20 tobacco genes characterizes four types of AC-elements. Further CsMYB4a-AC element and CsMYB4a-promoter interaction analyses indicate that the negative regulation of CsMYB4a on the shikimate and phenylpropanoid pathways in tobacco is via reducing promoter activity. Taken together, all data indicate that CsMYB4a negatively regulates the phenylpropanoid and shikimate pathways.Highlight: A tea (Camellia

  2. Molecular Cloning and Characterization of PnbHLH1 Transcription Factor in Panax notoginseng

    Directory of Open Access Journals (Sweden)

    Xiang Zhang

    2017-07-01

    Full Text Available Panax notoginseng has been extensively used as a traditional Chinese medicine. In the current study, molecular cloning and characterization of PnbHLH1 transcription factor were explored in Panax notoginseng. The full length of the PnbHLH1 gene obtained by splicing was 1430 bp, encoding 321 amino acids. Prokaryotic expression vector pET-28a-PnbHLH1 was constructed and transferred into the BL21 prokaryotic expression strain. An electrophoretic mobility shift assay of PnbHLH1 protein binding to E-box cis-acting elements verified that PnbHLH1 belonged to the bHLH class transcription factor which could interact with the promoter region of the E-box core sequence. The expression levels of key genes involved in the biosynthesis of triterpenoid saponins in PnbHLH1 transgenic cells were higher than those in the wild cells. Similarly, the total saponin contents were increased in the PnbHLH1 transgenic cell lines compared with the wild cell lines. Such results suggest that the PnbHLH1 transcription factor is a positive regulator in the biosynthesis of triterpenoid saponins in Panax notoginseng.

  3. Development of DNA affinity techniques for the functional characterization of purified RNA polymerase II transcription factors

    International Nuclear Information System (INIS)

    Garfinkel, S.; Thompson, J.A.; Cohen, R.B.; Brendler, T.; Safer, B.

    1987-01-01

    Affinity adsorption, precipitation, and partitioning techniques have been developed to purify and characterize RNA Pol II transcription components from whole cell extracts (WCE) (HeLa) and nuclear extracts (K562). The titration of these extracts with multicopy constructs of the Ad2 MLP but not pUC8, inhibits transcriptional activity. DNA-binding factors precipitated by this technique are greatly enriched by centrifugation. Using this approach, factors binding to the upstream promoter sequence (UPS) of the Ad2 MLP have been rapidly isolated by Mono Q, Mono S, and DNA affinity chromatography. By U.V. crosslinking to nucleotides containing specific 32 P-phosphodiester bonds within the recognition sequence, this factor is identified as a M/sub r/ = 45,000 polypeptide. To generate an assay system for the functional evaluation of single transcription components, a similar approach using synthetic oligonucleotide sequences spanning single promoter binding sites has been developed. The addition of a synthetic 63-mer containing the UPS element of the Ad2 MLP to HeLa WCE inhibited transcription by 60%. The addition of partially purified UPS binding protein, but not RNA Pol II, restored transcriptional activity. The addition of synthetic oligonucleotides containing other regulatory sequences not present in the Ad2 MLP was without effect

  4. Purification and characterization of factors produced by Aspergillus fumigatus which affect human ciliated respiratory epithelium.

    Science.gov (United States)

    Amitani, R; Taylor, G; Elezis, E N; Llewellyn-Jones, C; Mitchell, J; Kuze, F; Cole, P J; Wilson, R

    1995-09-01

    The mechanisms by which Aspergillus fumigatus colonizes the respiratory mucosa are unknown. Culture filtrates of eight of nine clinical isolates of A. fumigatus slowed ciliary beat frequency and damaged human respiratory epithelium in vitro. These changes appeared to occur concurrently. Culture filtrates of two clinical isolates of Candida albicans had no effect on ciliated epithelium. We have purified and characterized cilioinhibitory factors of a clinical isolate of A. fumigatus. The cilioinhibitory activity was heat labile, reduced by dialysis, and partially extractable into chloroform. The activity was associated with both high- and low-molecular-weight factors, as determined by gel filtration on Sephadex G-50. A low-molecular-weight cilioinhibitory factor was further purified by reverse-phase high-performance liquid chromatography and shown by mass spectrometry to be gliotoxin, a known metabolite of A. fumigatus. Gliotoxin significantly slowed ciliary beat frequency in association with epithelial damage at concentrations above 0.2 microgram/ml; other Aspergillus toxins, i.e., fumagillin and helvolic acid, were also cilioinhibitory but at much higher concentrations. High-molecular-weight (> or = 35,000 and 25,000) cilioinhibitory materials had neither elastolytic nor proteolytic activity and remain to be identified. Thus, A. fumigatus produces a number of biologically active substances which slow ciliary beating and damage epithelium and which may influence colonization of the airways.

  5. Corticotropin-releasing factor peptide antagonists: design, characterization and potential clinical relevance.

    Science.gov (United States)

    Rivier, Jean E; Rivier, Catherine L

    2014-04-01

    Elusive for more than half a century, corticotropin-releasing factor (CRF) was finally isolated and characterized in 1981 from ovine hypothalami and shortly thereafter, from rat brains. Thirty years later, much has been learned about the function and localization of CRF and related family members (Urocortins 1, 2 and 3) and their 2 receptors, CRF receptor type 1 (CRFR1) and CRF receptor type 2 (CRFR2). Here, we report the stepwise development of peptide CRF agonists and antagonists, which led to the CRFR1 agonist Stressin1; the long-acting antagonists Astressin2-B which is specific for CRFR2; and Astressin B, which binds to both CRFR1 and CRFR2.This analog has potential for the treatment of CRF-dependent diseases in the periphery, such as irritable bowel syndrome. Copyright © 2013 Elsevier Inc. All rights reserved.

  6. Land use impacts on biodiversity in LCA: proposal of characterization factors based on functional diversity

    DEFF Research Database (Denmark)

    Souza, Danielle Maia de; Flynn, Dan F. B.; DeClerck, Fabrice

    2013-01-01

    the distinctiveness of species; this paper proposes the use of an FD index to calculate characterization factors (CFs) for land use impacts. Furthermore, we compare the results of the CFs to current practice and assess the increase in complexity introduced by the use of the new indicator.The model proposed is based...... challenges. Such indices are data hungry (requiring species composition and traits) require more complex calculations than current common practice, including decisions on the choice of a method to calculate FD and the selection of traits.......The focus of land use modeling in life cycle impact assessment has been mainly on taxonomic measures of biodiversity, namely species richness (SR). However, increasing availability of trait data for species has led to the use of functional diversity (FD) as a promising metric to reflect...

  7. Human-factors-based implementation of the remote characterization system high-level control station

    International Nuclear Information System (INIS)

    Noakes, M.W.; Richardson, B.S.; Rowe, J.C.; Draper, J.V.; Sandness, G.R.

    1993-01-01

    The detection and characterization of buried objects and materials is an important first step in the restoration of the numerous US Department of Energy (DOE) and US Department of Defense waste disposal sites. DOE, through its Environmental Restoration and Waste Management Robotics and Technology Development Program, has developed the Remote Characterization System (RCS) to address the needs of remote subsurfacecharacterization. The RCS consists of a low-metal-content (low-metallic-signature) remotely piloted vehicle, a high-level control station (HLCS) where operators can remotely control the vehicle and analyze real-time data from sensors, and an array of sensors that can be chosen to meet the survey task at hand. Communication between the vehicle and the base station is handled by a radio link. Site mapping is made possible through the use of geopositioning satellite data. The primary mode of vehicle operation is teleoperation, but provision has been made for semiautonomous or supervisory control that allows for automated sitesurvey on simple sites. Data analysis and display is supported for both real-time observation and postprocessing of data. The particular emphasis of this paper documents the human-factors-based design influences on the HLCS and describes the design in detail

  8. Characterizing Stressors and Modifiable Health Risk Factors Among Homeless Smokers: An Exploratory Pilot Study.

    Science.gov (United States)

    Kendzor, Darla E; Reitzel, Lorraine R; Businelle, Michael S

    2015-10-01

    This pilot study was conducted to explore the associations between stressors related to homelessness and modifiable health risk factors (poor diet, insufficient physical activity, and overweight/obesity) and to provide direction for future research. Participants (N = 57) were homeless adults enrolled in a smoking cessation program. Analyses were conducted to characterize the sample as well as the relations between relevant stressors (discrimination, chronic stress, and fear and mistrust) and health risk factors. Inadequate daily consumption of fruits, vegetables, and fiber was common. High-fat diet and insufficient physical activity were also prevalent, and the majority of participants were overweight/obese. Participants commonly endorsed discrimination, fear of victimization, mistrust of others, and several other stressors. Greater endorsement of stressors was associated with a high-fat diet. Results suggest that lifestyle interventions and policy changes may be warranted in homeless shelters to attenuate the potential effects of stressors on high-fat dietary consumption among smokers. © 2015 Society for Public Health Education.

  9. Identification and network-enabled characterization of auxin response factor genes in Medicago truncatula

    Directory of Open Access Journals (Sweden)

    David J. Burks

    2016-12-01

    Full Text Available The Auxin Response Factor (ARF family of transcription factors is an important regulator of environmental response and symbiotic nodulation in the legume Medicago truncatula. While previous studies have identified members of this family, a recent spurt in gene expression data coupled with genome update and reannotation calls for a reassessment of the prevalence of ARF genes and their interaction networks in M. truncatula. We performed a comprehensive analysis of the M. truncatula genome and transcriptome that entailed search for novel ARF genes and the co-expression networks. Our investigation revealed 8 novel M. truncatula ARF (MtARF genes, of the total 22 identified, and uncovered novel gene co-expression networks as well. Furthermore, the topological clustering and single enrichment analysis of several network models revealed the roles of individual members of the MtARF family in nitrogen regulation, nodule initiation, and post-embryonic development through a specialized protein packaging and secretory pathway. In summary, this study not just shines new light on an important gene family, but also provides a guideline for identification of new members of gene families and their functional characterization through network analyses.

  10. Characterization and partial purification of Candida albicans Secretory IL-12 Inhibitory Factor

    Directory of Open Access Journals (Sweden)

    Chandra Jyotsna

    2008-02-01

    Full Text Available Abstract Background We have previously shown that supernatant from Candida albicans (CA culture contains a Secretory Interleukin (IL-12 Inhibitory Factor (CA-SIIF, which inhibits IL-12 production by human monocytes. However, the effect of CA-SIIF on secretion of other cytokines by monocytes is unknown, and detailed characterization of this factor has not been performed. Results In this study, we demonstrate that the IL-12 inhibitory activity of CA-SIIF was serum-independent, based on the reduction of IL-12 levels in monocytes stimulated under serum-independent conditions. The minimal inhibitory dose of CA-SIIF was found to be 200 μg/ml. Investigation of CA-SIIF's effect on macrophages IL-12 production in vitro and in vivo also showed that CA-SIIF inhibited IL-12 production by murine macrophages both in vitro (from 571 ± 24 pg/ml to 387 ± 87 pg/ml; P = 0.05 and in vivo (from 262 ± 6 pg/ml to 144 ± 30 pg/ml; P P P P Conclusion CA-SIIF is a glycoprotein which exhibits serum-independent inhibition of IL-12 production from monocytes in vitro and in vivo, and also modulates differentiation of monocytes into dendritic cells. These results suggest important role for CA-SIIF in interactions of C. albicans with the host immune system.

  11. Sector-wise midpoint characterization factors for impact assessment of regional consumptive and degradative water use.

    Science.gov (United States)

    Lin, Chia-Chun; Lin, Jia-Yu; Lee, Mengshan; Chiueh, Pei-Te

    2017-12-31

    Water availability, resulting from either a lack of water or poor water quality is a key factor contributing to regional water stress. This study proposes a set of sector-wise characterization factors (CFs), namely consumptive and degradative water stresses, to assess the impact of water withdrawals with a life cycle assessment approach. These CFs consider water availability, water quality, and competition for water between domestic, agricultural and industrial sectors and ecosystem at the watershed level. CFs were applied to a case study of regional water management of industrial water withdrawals in Taiwan to show that both regional or seasonal decrease in water availability contributes to a high consumptive water stress, whereas water scarcity due to degraded water quality not meeting sector standards has little influence on increased degradative water stress. Degradative water stress was observed more in the agricultural sector than in the industrial sector, which implies that the agriculture sector may have water quality concerns. Reducing water intensity and alleviating regional scale water stresses of watersheds are suggested as approaches to decrease the impact of both consumptive and degradative water use. The results from this study may enable a more detailed sector-wise analysis of water stress and influence water resource management policies. Copyright © 2017 Elsevier B.V. All rights reserved.

  12. Case Characterization, Clinical Features and Risk Factors in Drug-Induced Liver Injury

    Directory of Open Access Journals (Sweden)

    Aida Ortega-Alonso

    2016-05-01

    Full Text Available Idiosyncratic drug-induced liver injury (DILI caused by xenobiotics (drugs, herbals and dietary supplements presents with a range of both phenotypes and severity, from acute hepatitis indistinguishable of viral hepatitis to autoimmune syndromes, steatosis or rare chronic vascular syndromes, and from asymptomatic liver test abnormalities to acute liver failure. DILI pathogenesis is complex, depending on the interaction of drug physicochemical properties and host factors. The awareness of risk factors for DILI is arising from the analysis of large databases of DILI cases included in Registries and Consortia networks around the world. These networks are also enabling in-depth phenotyping with the identification of predictors for severe outcome, including acute liver failure and mortality/liver transplantation. Genome wide association studies taking advantage of these large cohorts have identified several alleles from the major histocompatibility complex system indicating a fundamental role of the adaptive immune system in DILI pathogenesis. Correct case definition and characterization is crucial for appropriate phenotyping, which in turn will strengthen sample collection for genotypic and future biomarkers studies.

  13. Characterization of responses and comparison of calibration factor for commercial MOSFET detectors.

    Science.gov (United States)

    Bharanidharan, Ganesan; Manigandan, Durai; Devan, Krishnamurthy; Subramani, Vellaiyan; Gopishankar, Natanasabapathi; Ganesh, Tharmar; Joshi, Rakeshchander; Rath, Gourakishore; Velmurugan, Jagadeesan; Aruna, Prakasarao; Ganesan, Singaravelu

    2005-01-01

    A commercial metal oxide silicon field effect transistor (MOSFET) dosimeter of model TN502-RD has been characterized for its linearity, reproducibility, field size dependency, dose rate dependency, and angular dependency for Cobalt-60 (60Co), 6-MV, and 15-MV beam energies. The performance of the MOSFET clearly shows that it is highly reproducible, independent of field size and dose rate. Furthermore, MOSFET has a very high degree of linearity, with r-value>0.9 for all 3 energies. The calibration factor for 2 similar MOSFET detectors of model TN502-RD were also estimated and compared for all 3 energies. The calibration factor between the 2 similar MOSFET detectors shows a variation of about 1.8% for 60Co and 15 MV, and for 6 MV it shows variation of about 2.5%, indicating that calibration should be done whenever a new MOSFET is used. However, the detector shows considerable angular dependency of about 8.8% variation. This may be due to the variation in radiation sensitivity between flat and bubble sides of the MOSFET, and indicates that positional care must be taken while using MOSFET for stereotactic radiosurgery and stereotactic radiotherapy dosimetric applications.

  14. Nanodiamond-based injectable hydrogel for sustained growth factor release: Preparation, characterization and in vitro analysis.

    Science.gov (United States)

    Pacelli, Settimio; Acosta, Francisca; Chakravarti, Aparna R; Samanta, Saheli G; Whitlow, Jonathan; Modaresi, Saman; Ahmed, Rafeeq P H; Rajasingh, Johnson; Paul, Arghya

    2017-08-01

    Nanodiamonds (NDs) represent an emerging class of carbon nanomaterials that possess favorable physical and chemical properties to be used as multifunctional carriers for a variety of bioactive molecules. Here we report the synthesis and characterization of a new injectable ND-based nanocomposite hydrogel which facilitates a controlled release of therapeutic molecules for regenerative applications. In particular, we have formulated a thermosensitive hydrogel using gelatin, chitosan and NDs that provides a sustained release of exogenous human vascular endothelial growth factor (VEGF) for wound healing applications. Addition of NDs improved the mechanical properties of the injectable hydrogels without affecting its thermosensitive gelation properties. Biocompatibility of the generated hydrogel was verified by in vitro assessment of apoptotic gene expressions and anti-inflammatory interleukin productions. NDs were complexed with VEGF and the inclusion of this complex in the hydrogel network enabled the sustained release of the angiogenic growth factor. These results suggest for the first time that NDs can be used to formulate a biocompatible, thermosensitive and multifunctional hydrogel platform that can function both as a filling agent to modulate hydrogel properties, as well as a delivery platform for the controlled release of bioactive molecules and growth factors. One of the major drawbacks associated with the use of conventional hydrogels as carriers of growth factors is their inability to control the release kinetics of the loaded molecules. In fact, in most cases, a burst release is inevitable leading to diminished therapeutic effects and unsuccessful therapies. As a potential solution to this issue, we hereby propose a strategy of incorporating ND complexes within an injectable hydrogel matrix. The functional groups on the surface of the NDs can establish interactions with the model growth factor VEGF and promote a prolonged release from the polymer network

  15. The origin of the Avram Iancu U-Ni-Co-Bi-As mineralization, Băiţa (Bihor) metallogenic district, Bihor Mts., Romania

    Science.gov (United States)

    Zajzon, Norbert; Szentpéteri, Krisztián; Szakáll, Sándor; Kristály, Ferenc

    2015-10-01

    The Băiţa metallogenic district in the Bihor Mountains is a historically important mining area in Romania. Uranium mining took place between 1952 and 1998 from various deposits, but very little is known about the geology and mineralogy of these deposits. In this paper, we describe geology and mineralogy of uranium mineralization of the Avram Iancu uranium mine from waste dump samples collected before complete remediation of the site. Texturally and mineralogically complex assemblages of nickeline, cobaltite-gersdorffite solid solution, native Bi, Bi-sulfosalts, molybdenite, and pyrite-chalcopyrite-sphalerite occur with uraninite, "pitchblende," and brannerite in most of the ore samples. The association of nickel, cobalt, and arsenic with uranium is reminiscent of five-element association of vein type U-Ni-Co-Bi-As deposits; however, the Avram Iancu ores appear to be more replacement-type stratiform/stratabound. Avram Iancu ore samples contain multistage complex, skarn, uranium sulfide, arsenide assemblages that can be interpreted to have been formed in the retrograde cooling stages of the skarn hydrothermal system. This mineralizing system may have built-up along Upper Cretaceous-Paleogene "Banatite" intrusions of diorite-to-granite composition. The intrusions crosscut the underlying uraniferous Permian formations in the stacked NW-verging Biharia Nappe System. The mineralization forms stacked, multilayer replacement horizons, along carbonate-rich lithologies within the metavolcanic (tuffaceous) Muncel Series. Mineral paragenesis and some mineral chemistry suggest moderate-to-high <450, i.e., 350-310 °C, formation temperatures for the uranium sulfide stage along stratigraphically controlled replacement zones and minor veins. Uranium minerals formed abundantly in this early stage and include botryoidal, sooty and euhedral uraninite, brannerite, and coffinite. Later and/or lower-temperature mineral assemblages include heterogeneous, complexly zoned arsenide

  16. U-Pb, Re-Os and Ar-Ar dating of the Linghou polymetallic deposit, Southeastern China: Implications for metallogenesis of the Qingzhou-Hangzhou metallogenic belt

    Science.gov (United States)

    Tang, Yanwen; Xie, Yuling; Liu, Liang; Lan, Tingguan; Yang, Jianling; Sebastien, Meffre; Yin, Rongchao; Liang, Songsong; Zhou, Limin

    2017-04-01

    The Qingzhou-Hangzhou metallogenic belt (QHMB) in Southeastern China has gained increasingly attention in recent years. However, due to the lack of reliable ages on intrusions and associated deposits in this belt, the tectonic setting and metallogenesis of the QHMB have not been well understood. The Linghou polymetallic deposit in northwestern Zhejiang Province is one of the typical deposits of the QHMB. According to the field relationships, this deposit consists of the early Cu-Au-Ag and the late Pb-Zn-Cu mineralization stages. Molybdenite samples with a mineral assemblage of molybdenite-chalcopyrite-pyrite ± quartz are collected from the copper mining tunnel near the Cu-Au-Ag ore bodies. Six molybdenite samples give the Re-Os model ages varying from 160.3 to 164.1 Ma and yield a mean age of 162.2 ± 1.4 Ma for the Cu-Au-Ag mineralization. Hydrothermal muscovite gives a well-defined Ar-Ar isochron age of 160.2 ± 1.1 Ma for the Pb-Zn-Cu mineralization. Three phases of granodioritic porphyry have been distinguished in this deposit, and LA-ICP-MS zircon U-Pb dating shows that they have formed at 158.8 ± 2.4 Ma, 158.3 ± 1.9 Ma and 160.6 ± 2.1 Ma, comparable to the obtained ages of the Cu-Au-Ag and Pb-Zn-Cu mineralization. Therefore, these intrusive rocks have a close temporal and spatial relationship with the Cu-Au-Ag and Pb-Zn-Cu ore bodies. The presences of skarn minerals (e.g., garnet) and vein-type ores, together with the previous fluid inclusion and H-O-C-S-Pb isotopic data, clearly indicate that the Cu-Au-Ag and Pb-Zn-Cu mineralization are genetically related to these granodiorite porphyries. This conclusion excludes the possibility that this deposit is of ;SEDEX; type and formed in a sag basin of continental rifts setting as previously proposed. Instead, it is proposed that the Linghou polymetallic and other similar deposits in the QHMB, such as the 150-160 Ma Yongping porphyry-skarn Cu-Mo, Dongxiang porphyry? Cu, Shuikoushan/Kangjiawang skarn Pb

  17. Dazai super-large uranium-bearing germanium deposit in western Yunnan region metallogenic geological conditions and prospect

    International Nuclear Information System (INIS)

    Han Yanrong; Yuan Qingbang; Li Yonghua; Zhang Ling; Dai Jiemin

    1995-05-01

    The Dazai super-large uranium-bearing germanium deposit is located in Bangmai Fault Basin, Western Yunnan, China. The basin basement is migmatitic granite and the cover is miocene coal-bearing clastics, Bangmai Formation. The basin development had undergone faulted rhombus basin forming, synsedimentary structure-developing and up-lifted-denuded stages. Synsedimentary faults had controlled distribution of sedimentary formation and lithofacies, and uranium and germanium mineralization. Germanium ore-bodies occur mainly in master lignite-bed of lower rhythmite. Hosted germanium-lignite is taken as main ore-type. Germanium occurs in vitrinite of lignite in the form of metal-organic complex. The metallogenetic geological conditions of the deposit are that ground preparation is uplift zone-migmatitic granite-fault basin-geothermal anomaly area, rich and thick ore-body is controlled by synsedimentary fault, peat-bog phase is favorable to accumulation for ore-forming elements, and unconformity between overlying cover and underlying basement is a channel-way of mineralizing fluid. A multiperiodic composite, being regarded sedimentation and diagenesis as a major process, uranium and germanium ore deposit has been formed through two mineralization. Four prospecting areas have been forecasted and two deposits have been accordingly discovered again. Technical-economic provableness shows that the deposit is characterized by shallow-buried, rich grade, large scale, easy mining and smelting. (9 figs.)

  18. Characterization factors for water footprint considering the scarcity of green and blue water sources

    Science.gov (United States)

    Oki, T.; Kondo, T.; Pokhrel, Y. N.; Hanasaki, N.

    2011-12-01

    The original concept of virtual water trade was invented to illustrate how much water demand can be reduced by importing food products (Allan 1996), and expanded for meat products and industrial products (Oki and Kanae, 2004). However, there was a confusion between "virtual trade of water" (original) and "trade of virtual water" (misinterpretation but widely accepted), and "virtual water" has been recognized as how much water was used to produce the commodity. Then, the concept has some analogy to carbon footprint (CFP) which is an indicator of total emission of greenhouse gases, and nowadays called water footprint (WFP, Hoekstra, 2004). However, WFP itself is just an inventory of water usages under the framework of life cycle assessment (LCA), and the volume of WFP does not necessary reflect the environmental impacts of water usages because consumptive water use of 100 liter from ground water in arid regions just before rainy season would have more environmental impacts than consumptive water use of 100 liter from rain water in humid regions during snow melt season. In the case of CFP, the emissions of five greenhouse gases except for CO2 were converted into CO2 equivalent volumes by considering the sensitivity for the global warming potential, and summed up into CFP. Here, we propose a new idea objectively determining the weights (characterization factors) for blue water usages, such as from river and ground water, to be converted into green water equivalent in each region and time. The weights are inversely proportional to the area required to obtain the same amount of green water, and water balance model can provide the basic information. The new concept was applied to the WFP of Japan through the imports of major crops. As an inventory, WFP was 15.5 km3/y of rain water, 2.2 km3/y of river water, and 2.0 km3/y of non-renewable and non-local water (NRNL water) for year 2000, however, considering the proposed characterization factors in each region (0.5 x 0

  19. Phenotypic and genotypic characterization of four factor VII deficiency patients from central China.

    Science.gov (United States)

    Liu, Hui; Wang, Hua-Fang; Cheng, Zhi-peng; Wang, Qing-yun; Hu, Bei; Zeng, Wei; Wu, Ying-ying; Guo, Tao; Tang, Liang; Hu, Yu

    2015-06-01

    Hereditary coagulation factor VII deficiency (FVIID) is a rare autosomal, recessive inherited hemorrhagic disorder related to a variety of mutations or polymorphisms throughout the factor VII (FVII) gene (F7). The aims of this study were to characterize the molecular defect of the F7 gene in four unrelated patients with FVIID and to find the genotype-phenotype correlation. All nine exons, exon-intron boundaries, and 5' and 3'-untranslated regions of the F7 gene were amplified by PCR and the purified PCR products were sequenced directly. Suspected mutations were confirmed by another PCR and sequencing of the opposite strand. Family studies were also performed. A total of five unique lesions were identified, including three missense mutations (c.384A>G, c.839A>C, c.1163T>G, predicting p.Tyr128Cys, p.Glu280Ala and p.Phe388Cys substitution, respectively) and two splice junction mutations (c.572-1G>A, c.681+1G>T), among which two (p.Glu280Ala, p.Phe388Cys) were novel. A previously reported mutation p.Tyr128Cys was seen in the homozygous state in two unrelated patients. The other two cases were both compound heterozygotes of a missense mutation and a splicing site mutation. Multiple sequence alignment using DNAMAN analysis showed that all the missense mutations were found in residues that highly conserved across species and vitamin K-dependent serine proteases. Online software Polyphen and SIFT were used to confirm the pathogenic of the missense mutation. p.Tyr128Cys seems to be a hotspot of the F7 gene in ethnic Han Chinese population.

  20. Characterization of mechanical behavior of an epithelial monolayer in response to epidermal growth factor stimulation

    International Nuclear Information System (INIS)

    Yang, Ruiguo; Chen, Jennifer Y.; Xi, Ning; Lai, King Wai Chiu; Qu, Chengeng; Fung, Carmen Kar Man; Penn, Lynn S.; Xi, Jun

    2012-01-01

    Cell signaling often causes changes in cellular mechanical properties. Knowledge of such changes can ultimately lead to insight into the complex network of cell signaling. In the current study, we employed a combination of atomic force microscopy (AFM) and quartz crystal microbalance with dissipation monitoring (QCM-D) to characterize the mechanical behavior of A431 cells in response to epidermal growth factor receptor (EGFR) signaling. From AFM, which probes the upper portion of an individual cell in a monolayer of cells, we observed increases in energy dissipation, Young's modulus, and hysteresivity. Increases in hysteresivity imply a shift toward a more fluid-like mechanical ordering state in the bodies of the cells. From QCM-D, which probes the basal area of the monolayer of cells collectively, we observed decreases in energy dissipation factor. This result suggests a shift toward a more solid-like state in the basal areas of the cells. The comparative analysis of these results indicates a regionally specific mechanical behavior of the cell in response to EGFR signaling and suggests a correlation between the time-dependent mechanical responses and the dynamic process of EGFR signaling. This study also demonstrates that a combination of AFM and QCM-D is able to provide a more complete and refined mechanical profile of the cells during cell signaling. -- Highlights: ► The EGF-induced cellular mechanical response is regionally specific. ► The EGF-induced cellular mechanical response is time and dose dependent. ► A combination of AFM and QCM-D provides a more complete mechanical profile of cells.

  1. Nanotoxicity and Life Cycle Assessment: First attempt towards the determination of characterization factors for carbon nanotubes

    Science.gov (United States)

    Rodriguez-Garcia, Gonzalo; Zimmermann, Benedikt; Weil, Marcel

    2014-08-01

    Carbon materials, whether at macro, micro or at nanoscale, play an important role in the battery industry, as they can be used as electrodes, electrode enhancers, bipolar separators, or current collectors. When conducting a Life Cycle Assessment (LCA) of novel batteries manufacturing processes, we also need to consider the fate of potentially emitted carbon based nanomaterials. However, the knowledge generated in the last decade regarding the behavior of such materials in the environment and its toxicological effects has yet to be included in the Life Cycle Impact Assessment (LCIA) methodologies. Conventional databases of chemical products (e.g. ECHA, ECOTOX) offer little information regarding engineered nanomaterials (ENM). It is thus necessary to go one step further and compile physicochemical and toxicological data directly from scientific literature. Such studies do not only differ in their results, but also in their methodologies, and several calls have been made towards a more consistent approach that would allow us model the fate of ENM in the environment as well as their potentially harmful effects. Trying to overcome these limitations we have developed a tool based on Microsoft Excel® combining several methods for the estimation of physicochemical properties of carbon nanotubes (CNT). The information generated with this tool is combined with degradation rates and toxicological data consistent with the methods followed by the USEtox methodology. Thus, it is possible to calculate the characterization factors of CNTs and integrate them as a first proxy in future LCA of products including these ENM.

  2. Characterization of insulin-like growth factor I produced by fetal rat pancreatic islets

    International Nuclear Information System (INIS)

    Scharfmann, R.; Corvol, M.; Czernichow, P.

    1989-01-01

    Pancreatic islets were prepared from 22-day-old rat fetuses. After 5 days of culture in dishes allowing cell attachment, neoformed islets were kept free floating in RPMI-1640 medium (16.5 mM glucose, 1% fetal calf serum). The islets were then pulsed with [ 3 H]leucine and [ 35 S]methionine for 24 h. The conditioned medium was acidified with acetic acid (final pH 2.7), desalted, concentrated, and gel filtered on Bio-Gel P100 in acid conditions. The radioactive material that comigrated with immunoreactive insulinlike growth factor I (IGF-I) produced by the islets was pooled, concentrated, and further characterized by reverse-phase high-performance liquid chromatography on a C18 Bondapak column with a linear gradient of acetonitrile (20-80%). The radioactive material that eluted as pure IGF-I (40% acetonitrile) was further studied by chromatofocusing on a Pharmacia PBE 94 column. A sharp radioactive peak containing [ 3 H]leucine and [ 35 S]methionine was eluted at pH 8.55. This material was immunoprecipitated with an antiserum to IGF-I. This study demonstrated that fetal islet cells synthesize molecules that are, by several criteria, equivalent to native IGF-I

  3. Family-wide characterization of the DENN domain Rab GDP-GTP exchange factors.

    Science.gov (United States)

    Yoshimura, Shin-ichiro; Gerondopoulos, Andreas; Linford, Andrea; Rigden, Daniel J; Barr, Francis A

    2010-10-18

    A key requirement for Rab function in membrane trafficking is site-specific activation by GDP-GTP exchange factors (GEFs), but the majority of the 63 human Rabs have no known GEF. We have performed a systematic characterization of the 17 human DENN domain proteins and demonstrated that they are specific GEFs for 10 Rabs. DENND1A/1B localize to clathrin patches at the plasma membrane and activate Rab35 in an endocytic pathway trafficking Shiga toxin to the trans-Golgi network. DENND2 GEFs target to actin filaments and control Rab9-dependent trafficking of mannose-6-phosphate receptor to lysosomes. DENND4 GEFs target to a tubular membrane compartment adjacent to the Golgi, where they activate Rab10, which suggests a function in basolateral polarized sorting in epithelial cells that compliments the non-DENN GEF Sec2 acting on Rab8 in apical sorting. DENND1C, DENND3, DENND5A/5B, MTMR5/13, and MADD activate Rab13, Rab12, Rab39, Rab28, and Rab27A/27B, respectively. Together, these findings provide a basis for future studies on Rab regulation and function.

  4. Characterization of Bombyx mori mitochondrial transcription factor A, a conserved regulator of mitochondrial DNA.

    Science.gov (United States)

    Sumitani, Megumi; Kondo, Mari; Kasashima, Katsumi; Endo, Hitoshi; Nakamura, Kaoru; Misawa, Toshihiko; Tanaka, Hiromitsu; Sezutsu, Hideki

    2017-04-15

    In the present study, we initially cloned and characterized a mitochondrial transcription factor A (Tfam) homologue in the silkworm, Bombyx mori. Bombyx mori TFAM (BmTFAM) localized to mitochondria in cultured silkworm and human cells, and co-localized with mtDNA nucleoids in human HeLa cells. In an immunoprecipitation analysis, BmTFAM was found to associate with human mtDNA in mitochondria, indicating its feature as a non-specific DNA-binding protein. In spite of the low identity between BmTFAM and human TFAM (26.5%), the expression of BmTFAM rescued mtDNA copy number reductions and enlarged mtDNA nucleoids in HeLa cells, which were induced by human Tfam knockdown. Thus, BmTFAM compensates for the function of human TFAM in HeLa cells, demonstrating that the mitochondrial function of TFAM is highly conserved between silkworms and humans. BmTfam mRNA was strongly expressed in early embryos. Through double-stranded RNA (dsRNA)-based RNA interference (RNAi) in silkworm embryos, we found that the knockdown of BmTFAM reduced the amount of mtDNA and induced growth retardation at the larval stage. Collectively, these results demonstrate that BmTFAM is a highly conserved mtDNA regulator and may be a good candidate for investigating and modulating mtDNA metabolism in this model organism. Copyright © 2017 Elsevier B.V. All rights reserved.

  5. Characterization of an apparently synonymous F5 mutation causing aberrant splicing and factor V deficiency.

    Science.gov (United States)

    Nuzzo, F; Bulato, C; Nielsen, B I; Lee, K; Wielders, S J; Simioni, P; Key, N S; Castoldi, E

    2015-03-01

    Coagulation factor V (FV) deficiency is a rare autosomal recessive bleeding disorder. We investigated a patient with severe FV deficiency (FV:C mutation in exon 4 (c.578G>C, p.Cys193Ser), predicting the abolition of a conserved disulphide bridge, and an apparently synonymous variant in exon 8 (c.1281C>G). The observation that half of the patient's F5 mRNA lacked the last 18 nucleotides of exon 8 prompted us to re-evaluate the c.1281C>G variant for its possible effects on splicing. Bioinformatics sequence analysis predicted that this transversion would activate a cryptic donor splice site and abolish an exonic splicing enhancer. Characterization in a F5 minigene model confirmed that the c.1281C>G variant was responsible for the patient's splicing defect, which could be partially corrected by a mutation-specific morpholino antisense oligonucleotide. The aberrantly spliced F5 mRNA, whose stability was similar to that of the normal mRNA, encoded a putative FV mutant lacking amino acids 427-432. Expression in COS-1 cells indicated that the mutant protein is poorly secreted and not functional. In conclusion, the c.1281C>G mutation, which was predicted to be translationally silent and hence neutral, causes FV deficiency by impairing pre-mRNA splicing. This finding underscores the importance of cDNA analysis for the correct assessment of exonic mutations. © 2014 John Wiley & Sons Ltd.

  6. Characterization of the receptors for mycobacterial cord factor in Guinea pig.

    Directory of Open Access Journals (Sweden)

    Kenji Toyonaga

    Full Text Available Guinea pig is a widely used animal for research and development of tuberculosis vaccines, since its pathological disease process is similar to that present in humans. We have previously reported that two C-type lectin receptors, Mincle (macrophage inducible C-type lectin, also called Clec4e and MCL (macrophage C-type lectin, also called Clec4d, recognize the mycobacterial cord factor, trehalose-6,6'-dimycolate (TDM. Here, we characterized the function of the guinea pig homologue of Mincle (gpMincle and MCL (gpMCL. gpMincle directly bound to TDM and transduced an activating signal through ITAM-bearing adaptor molecule, FcRγ. Whereas, gpMCL lacked C-terminus and failed to bind to TDM. mRNA expression of gpMincle was detected in the spleen, lymph nodes and peritoneal macrophages and it was strongly up-regulated upon stimulation of zymosan and TDM. The surface expression of gpMincle was detected on activated macrophages by a newly established monoclonal antibody that also possesses a blocking activity. This antibody potently suppressed TNF production in BCG-infected macrophages. Collectively, gpMincle is the TDM receptor in the guinea pig and TDM-Mincle axis is involved in host immune responses against mycobacteria.

  7. Characterization of the receptors for mycobacterial cord factor in Guinea pig.

    Science.gov (United States)

    Toyonaga, Kenji; Miyake, Yasunobu; Yamasaki, Sho

    2014-01-01

    Guinea pig is a widely used animal for research and development of tuberculosis vaccines, since its pathological disease process is similar to that present in humans. We have previously reported that two C-type lectin receptors, Mincle (macrophage inducible C-type lectin, also called Clec4e) and MCL (macrophage C-type lectin, also called Clec4d), recognize the mycobacterial cord factor, trehalose-6,6'-dimycolate (TDM). Here, we characterized the function of the guinea pig homologue of Mincle (gpMincle) and MCL (gpMCL). gpMincle directly bound to TDM and transduced an activating signal through ITAM-bearing adaptor molecule, FcRγ. Whereas, gpMCL lacked C-terminus and failed to bind to TDM. mRNA expression of gpMincle was detected in the spleen, lymph nodes and peritoneal macrophages and it was strongly up-regulated upon stimulation of zymosan and TDM. The surface expression of gpMincle was detected on activated macrophages by a newly established monoclonal antibody that also possesses a blocking activity. This antibody potently suppressed TNF production in BCG-infected macrophages. Collectively, gpMincle is the TDM receptor in the guinea pig and TDM-Mincle axis is involved in host immune responses against mycobacteria.

  8. Nanotoxicity and Life Cycle Assessment: First attempt towards the determination of characterization factors for carbon nanotubes

    International Nuclear Information System (INIS)

    Rodriguez-Garcia, Gonzalo; Weil, Marcel; Zimmermann, Benedikt

    2014-01-01

    Carbon materials, whether at macro, micro or at nanoscale, play an important role in the battery industry, as they can be used as electrodes, electrode enhancers, bipolar separators, or current collectors. When conducting a Life Cycle Assessment (LCA) of novel batteries manufacturing processes, we also need to consider the fate of potentially emitted carbon based nanomaterials. However, the knowledge generated in the last decade regarding the behavior of such materials in the environment and its toxicological effects has yet to be included in the Life Cycle Impact Assessment (LCIA) methodologies. Conventional databases of chemical products (e.g. ECHA, ECOTOX) offer little information regarding engineered nanomaterials (ENM). It is thus necessary to go one step further and compile physicochemical and toxicological data directly from scientific literature. Such studies do not only differ in their results, but also in their methodologies, and several calls have been made towards a more consistent approach that would allow us model the fate of ENM in the environment as well as their potentially harmful effects. Trying to overcome these limitations we have developed a tool based on Microsoft Excel ® combining several methods for the estimation of physicochemical properties of carbon nanotubes (CNT). The information generated with this tool is combined with degradation rates and toxicological data consistent with the methods followed by the USEtox methodology. Thus, it is possible to calculate the characterization factors of CNTs and integrate them as a first proxy in future LCA of products including these ENM

  9. Characterization of cell-surface receptors for monoclonal-nonspecific suppressor factor (MNSF)

    International Nuclear Information System (INIS)

    Nakamura, M.; Ogawa, H.; Tsunematsu, T.

    1990-01-01

    Monoclonal-nonspecific suppressor factor (MNSF) is a lymphokine derived from murine T cell hybridoma. The target tissues are both LPS-stimulated B cells and Con A-stimulated T cells. Since the action of MNSF may be mediated by its binding to specific cell surface receptors, we characterized the mode of this binding. The purified MNSF was labeled with 125 I, using the Bolton-Hunter reagent. The labeled MNSF bound specifically to a single class of receptor (300 receptors per cell) on mitogen-stimulated murine B cells or T cells with an affinity of 16 pM at 24 degrees C, in the presence of sodium azide. Competitive experiments showed that MNSF bound to the specific receptor and that the binding was not shared with IL2, IFN-gamma, and TNF. Various cell types were surveyed for the capacity to specifically bind 125 I-MNSF. 125 I-MNSF bound to MOPC-31C (a murine plasmacytoma line) and to EL4 (a murine T lymphoma line). The presence of specific binding correlates with the capacity of the cells to respond to MNSF. These data support the view that like other polypeptide hormones, the action of MNSF is mediated by specific cell surface membrane receptor protein. Identification of these receptors will provide insight into the apparently diverse activities of MNSF

  10. Expression of human blood coagulation factor XI: characterization of the defect in factor XI type III deficiency

    NARCIS (Netherlands)

    Meijers, J. C.; Davie, E. W.; Chung, D. W.

    1992-01-01

    Human factor XI (FXI) is a blood coagulation factor participating in the early phase of the intrinsic pathway of blood coagulation. It circulates in blood as a glycoprotein composed of two identical chains held together by a single disulfide bond between the fourth apple domains. FXI has been

  11. Genome-wide identification of the regulatory targets of a transcription factor using biochemical characterization and computational genomic analysis

    Directory of Open Access Journals (Sweden)

    Jolly Emmitt R

    2005-11-01

    Full Text Available Abstract Background A major challenge in computational genomics is the development of methodologies that allow accurate genome-wide prediction of the regulatory targets of a transcription factor. We present a method for target identification that combines experimental characterization of binding requirements with computational genomic analysis. Results Our method identified potential target genes of the transcription factor Ndt80, a key transcriptional regulator involved in yeast sporulation, using the combined information of binding affinity, positional distribution, and conservation of the binding sites across multiple species. We have also developed a mathematical approach to compute the false positive rate and the total number of targets in the genome based on the multiple selection criteria. Conclusion We have shown that combining biochemical characterization and computational genomic analysis leads to accurate identification of the genome-wide targets of a transcription factor. The method can be extended to other transcription factors and can complement other genomic approaches to transcriptional regulation.

  12. Characterization factors for terrestrial acidification at the global scale: a systematic analysis of spatial variability and uncertainty.

    Science.gov (United States)

    Roy, Pierre-Olivier; Azevedo, Ligia B; Margni, Manuele; van Zelm, Rosalie; Deschênes, Louise; Huijbregts, Mark A J

    2014-12-01

    Characterization factors (CFs) are used in life cycle assessment (LCA) to quantify the potential impact per unit of emission. CFs are obtained from a characterization model which assess the environmental mechanisms along the cause-effect chain linking an emission to its potential damage on a given area of protection, such as loss in ecosystem quality. Up to now, CFs for acidifying emissions did not cover the global scale and were only representative of their characterization model geographical scope. Consequently, current LCA practices implicitly assume that all emissions from a global supply chain occur within the continent referring to the characterization method geographical scope. This paper provides worldwide 2°×2.5° spatially-explicit CFs, representing the change in relative loss of terrestrial vascular plant species due to an emission change of nitrogen oxides (NOx), ammonia (NH3) and sulfur dioxide (SO2). We found that spatial variability in the CFs is much larger compared to statistical uncertainty (six orders of magnitude vs. two orders of magnitude). Spatial variability is mainly caused by the atmospheric fate factor and soil sensitivity factor, while the ecological effect factor is the dominant contributor to the statistical uncertainty. The CFs provided in our study allow the worldwide spatially explicit evaluation of life cycle impacts related to acidifying emissions. This opens the door to evaluate regional life cycle emissions of different products in a global economy. Copyright © 2014 Elsevier B.V. All rights reserved.

  13. Characterization of connective tissue growth factor expression in primary cultures of human tubular epithelial cells: modulation by hypoxia

    NARCIS (Netherlands)

    Kroening, Sven; Neubauer, Emily; Wullich, Bernd; Aten, Jan; Goppelt-Struebe, Margarete

    2010-01-01

    Kroening S, Neubauer E, Wullich B, Aten J, Goppelt-Struebe M. Characterization of connective tissue growth factor expression in primary cultures of human tubular epithelial cells: modulation by hypoxia. Am J Physiol Renal Physiol 298:F796-F806, 2010. First published December 23, 2009;

  14. Structural, functional and evolutionary characterization of major drought transcription factors families in maize

    Science.gov (United States)

    Mittal, Shikha; Banduni, Pooja; Mallikarjuna, Mallana G.; Rao, Atmakuri R.; Jain, Prashant A.; Dash, Prasanta K.; Thirunavukkarasu, Nepolean

    2018-05-01

    Drought is one of the major threats to maize production. In order to improve the production and to breed tolerant hybrids, understanding the genes and regulatory mechanisms during drought stress is important. Transcription factors (TFs) play a major role in gene regulation and many TFs have been identified in response to drought stress. In our experiment, a set of 15 major TF families comprising 1436 genes was structurally and functionally characterized using in-silico tools and a gene expression assay. All 1436 genes were mapped on 10 chromosome of maize. The functional annotation indicated the involvement of these genes in ABA signaling, ROS scavenging, photosynthesis, stomatal regulation, and sucrose metabolism. Duplication was identified as the primary force in divergence and expansion of TF families. Phylogenetic relationship was developed individually for each TF family as well as combined TF families. Phylogenetic analysis grouped the TF family of genes into TF-specific and mixed groups. Phylogenetic analysis of genes belonging to various TF families suggested that the origin of TFs occurred in the lineage of maize evolution. Gene structure analysis revealed that more number of genes were intron-rich as compared to intronless genes. Drought-responsive CRE’s such as ABREA, ABREB, DRE1 and DRECRTCOREAT have been identified. Expression and interaction analyses identified leaf-specific bZIP TF, GRMZM2G140355, as a potential contributor toward drought tolerance in maize. We also analyzed protein-protein interaction network of 269 drought-responsive genes belonging to different drought-related TFs. The information generated on structural and functional characteristics, expression and interaction of the drought-related TF families will be useful to decipher the drought tolerance mechanisms and to derive drought-tolerant genotypes in maize.

  15. Use of geographic indicators of healthcare, environment and socioeconomic factors to characterize environmental health disparities.

    Science.gov (United States)

    Padilla, Cindy M; Kihal-Talantikit, Wahida; Perez, Sandra; Deguen, Severine

    2016-07-22

    An environmental health inequality is a major public health concern in Europe. However just few studies take into account a large set of characteristics to analyze this problematic. The aim of this study was to identify and describe how socioeconomic, health accessibility and exposure factors accumulate and interact in small areas in a French urban context, to assess environmental health inequalities related to infant and neonatal mortality. Environmental indicators on deprivation index, proximity to high-traffic roads, green space, and healthcare accessibility were created using the Geographical Information System. Cases were collected from death certificates in the city hall of each municipality in the Nice metropolitan area. Using the parental addresses, cases were geocoded to their census block of residence. A classification using a Multiple Component Analysis following by a Hierarchical Clustering allow us to characterize the census blocks in terms of level of socioeconomic, environmental and accessibility to healthcare, which are very diverse definition by nature. Relation between infant and neonatal mortality rate and the three environmental patterns which categorize the census blocks after the classification was performed using a standard Poisson regression model for count data after checking the assumption of dispersion. Based on geographic indicators, three environmental patterns were identified. We found environmental inequalities and social health inequalities in Nice metropolitan area. Moreover these inequalities are counterbalance by the close proximity of deprived census blocks to healthcare facilities related to mother and newborn. So therefore we demonstrate no environmental health inequalities related to infant and neonatal mortality. Examination of patterns of social, environmental and in relation with healthcare access is useful to identify census blocks with needs and their effects on health. Similar analyzes could be implemented and considered

  16. Characterization of putative virulence factors of Serratia marcescens strain SEN for pathogenesis in Spodoptera litura.

    Science.gov (United States)

    Aggarwal, Chetana; Paul, Sangeeta; Tripathi, Vishwas; Paul, Bishwajeet; Khan, Md Aslam

    2017-02-01

    Two Serratia marcescens strains, SEN and ICC-4, isolated from diseased insect cadavers were observed to differ considerably in their virulence towards Spodoptera litura. The present study was aimed to characterize the possible virulence factors present in the virulent Serratia marcescens strain SEN. Both the S. marcescens strains were evaluated for the presence of various lytic enzymes such as chitinase, lipase, protease and phospholipase. The virulent S. marcescens strain SEN was observed to possess considerably higher activity of chitinase and protease enzymes; activity of phospholipase enzyme was also higher. Although, all the three toxin genes shlA, phlA and swr could be detected in both the S. marcescens strains, there was a higher expression of these genes in the virulent strain SEN. S. marcescens strain ICC-4 showed greater reduction in overall growth yield in the post-exponential phase in the presence of midgut juice and hemolymph of S. litura larvae, as compared to S. marcescens strain SEN. Proliferation of the S. marcescens strain SEN was also considerably higher in foregut, midgut and hemolymph of S. litura larvae, as compared to strain ICC-4. Peritrophic membrane treated with broth culture of the S. marcescens strain SEN showed higher damage as compared to strain ICC-4. The peritrophic membrane of larvae fed on diet treated with the virulent strain showed considerable damage while the peritrophic membrane of larvae fed on diet treated with the non-virulent strain showed no damage. This is the first report documenting the fate of ingested S. marcescens in S. litura gut and the relative expression of toxin genes from two S. marcescens strains differing in their virulence towards S. litura. Copyright © 2016 Elsevier Inc. All rights reserved.

  17. Characterization of the epidermal growth factor receptor associated with cytoskeletons of A431 cells

    International Nuclear Information System (INIS)

    Roy, L.M.; Gittinger, C.K.; Landreth, G.E.

    1989-01-01

    Epidermal growth factor receptors (EGF-R) have been shown to be associated with the detergent-insoluble cytoskeleton of A431 cells, where they retained both a functional ligand-binding domain and tyrosine kinase activity. In the present study we have characterized the tyrosine kinase and ligand binding activities of this cytoskeletally associated EGF-R. The tyrosine kinase activity of the cytoskeletally associated EGF-R was stimulated by EGF treatment of intact cells as evidenced by increased autophosphorylation and phosphorylation of the exogenous substrate angiotensin II (AII). The kinetic behavior of the EGF-R associated with cytoskeletons of EGF-treated cells was similar to that of purified receptors. The stimulation of the receptor kinase activity required EGF treatment of intact cells prior to Triton extraction. If cytoskeletons were prepared from untreated cells and then incubated with EGF, there was no stimulation of the detergent-insoluble receptor kinase activity, indicating that the immobilized receptor was unable to undergo EGF-stimulated activation. Comparison of peptide maps from soluble and cytoskeletally associated EGF-R revealed qualitatively similar patterns; however, they are distinguished by a prominent 46 kD band in digests of the cytoskeletal EGF-R. Saturable binding of 125I-EGF to A431 cytoskeletons prepared from adherent and suspended cells demonstrated the presence of specific receptors on the cytoskeleton. High-affinity EGF-R were preferentially retained upon detergent extraction of adherent cells, whereas both low- and high-affinity receptors were solubilized from the cytoskeletons of suspended cells. Suspension of cells resulted in the solubilization of an additional 15% of the EGF-R to that solubilized in adherent cells, indicating that EGF-R can reversibly associate with the structural elements of the cell

  18. Molecular characterization of Quercus suber MYB1, a transcription factor up-regulated in cork tissues.

    Science.gov (United States)

    Almeida, Tânia; Menéndez, Esther; Capote, Tiago; Ribeiro, Teresa; Santos, Conceição; Gonçalves, Sónia

    2013-01-15

    The molecular processes associated with cork development in Quercus suber L. are poorly understood. A previous molecular approach identified a list of genes potentially important for cork formation and differentiation, providing a new basis for further molecular studies. This report is the first molecular characterization of one of these candidate genes, QsMYB1, coding for an R2R3-MYB transcription factor. The R2R3-MYB gene sub-family has been described as being involved in the phenylpropanoid and lignin pathways, both involved in cork biosynthesis. The results showed that the expression of QsMYB1 is putatively mediated by an alternative splicing (AS) mechanism that originates two different transcripts (QsMYB1.1 and QsMYB1.2), differing only in the 5'-untranslated region, due to retention of the first intron in one of the variants. Moreover, within the retained intron, a simple sequence repeat (SSR) was identified. The upstream regulatory region of QsMYB1 was extended by a genome walking approach, which allowed the identification of the putative gene promoter region. The relative expression pattern of QsMYB1 transcripts determined by reverse transcription quantitative polymerase chain reaction (RT-qPCR) revealed that both transcripts were up-regulated in cork tissues; the detected expression was several times higher in newly formed cork harvested from trees producing virgin, second or reproduction cork when compared with wood. Moreover, the expression analysis of QsMYB1 in several Q. suber organs showed very low expression in young branches and roots, whereas in leaves, immature acorns or male flowers, no expression was detected. These preliminary results suggest that QsMYB1 may be related to secondary growth and, in particular, with the cork biosynthesis process with a possible alternative splicing mechanism associated with its regulatory function. Copyright © 2012 Elsevier GmbH. All rights reserved.

  19. Functional Characterization of the Maize Phytochrome-Interacting Factors PIF4 and PIF5

    Directory of Open Access Journals (Sweden)

    Qingbiao Shi

    2018-01-01

    Full Text Available Phytochrome-interacting factors (PIFs play important roles in photomorphogenesis, the shade avoidance response, and other aspects of plant growth and development. PIF family proteins have been well-studied in Arabidopsis thaliana, but little is known about their physiological functions and molecular mechanisms in maize (Zea mays. In this study, we investigated the physiological functions of ZmPIF4 and ZmPIF5, two highly conserved members of the PIF gene family. RT-qPCR and western blot analyses revealed that ZmPIF4 and ZmPIF5 expression and ZmPIF4 and ZmPIF5 levels peak at night and remain low during the day. Overexpression of ZmPIF4 and ZmPIF5 in Arabidopsis partially rescued the reduced hypocotyl elongation and defective response to gravity in pif1 pif3 pif4 pif5 quadruple mutants (pifq. In addition, under high red: far-red light conditions, Arabidopsis lines overexpressing ZmPIF4 exhibited a constitutive shade avoidance response, including early flowering, slender leaves and inflorescences, plant lodging and precocious leaf senescence. Furthermore, ZmPIF4 physically interacted with the Arabidopsis DELLA protein REPRESSOR OF GA1-3 (RGA, indicating a potential interaction between ZmPIF4 and gibberellin signaling pathway on plant growth. Taken together, our results revealed that ZmPIF4 and ZmPIF5 are functionally conserved proteins that may play conserved roles in the response to phytochrome signaling in plants.Highlights:In this study, the functions of ZmPIF4 and ZmPIF5 were characterized by expression in Arabidopsis, revealing conserved roles of PIF family proteins in photomorphogenesis and the shade avoidance response in land plants.

  20. CARACTERIZACIÓN DE LOS FACTORES DE RIESGO VASCULAR EN PACIENTES ADULTOS / Characterization of vascular risk factors in adult patients

    OpenAIRE

    Estrella M. Hernández Hurtado; Yannelis Rodríguez Valido; Valentina Carpio García; Isis Yera Alós; Liset Valdés Manresa; Nercy O. Díaz Águila; Orestes Díaz Castro; Orestes Díaz Águila

    2013-01-01

    Resumen: Introducción: La elevada prevalencia de factores de riesgo vascular en la población y su relación con el desarrollo de la cardiopatía isquémica representan un problema de salud pública. Objetivo: Caracterizar estos factores en adultos. Método: Se realizó un estudio observacional, descriptivo, transversal en pacientes que asistieron a la consulta de riesgo vascular del municipio de Ranchuelo, en Villa Clara, entre abril de 2010 y abril de 2011. La información se procesó con el program...

  1. Phenylpropanoids accumulation in eggplant fruit: characterization of biosynthetic genes and regulation by a MYB transcription factor

    Directory of Open Access Journals (Sweden)

    Teresa eDocimo

    2016-01-01

    Full Text Available Phenylpropanoids are major secondary metabolites in eggplant (Solanum melongena fruits. Chlorogenic acid (CGA accounts for 70 to 90% of total phenolics in flesh tissues, while anthocyanins are mainly present in the fruit skin. As a contribution to the understanding of the peculiar accumulation of these health-promoting metabolites in eggplant, we report on metabolite abundance, regulation of CGA and anthocyanin biosynthesis, and characterization of candidate CGA biosynthetic genes in S. melongena.Higher contents of CGA, Delphinidin 3-rutinoside and rutin were found in eggplant fruits compared to other tissues, associated to an elevated transcript abundance of structural genes such as PAL, HQT, DFR and ANS, suggesting that active in situ biosynthesis contributes to anthocyanin and CGA accumulation in fruit tissues. Putative orthologs of the two CGA biosynthetic genes PAL and HQT, as well as a variant of a MYB1 transcription factor showing identity with group 6 MYBs, were isolated from an Occidental S. melongena traditional variety and demonstrated to differ from published sequences from Asiatic varieties.In silico analysis of the isolated SmPAL1, SmHQT1, SmANS, and SmMyb1 promoters revealed the presence of several Myb regulatory elements for the biosynthetic genes and unique elements for the TF, suggesting its involvement in other physiological roles beside phenylpropanoid biosynthesis regulation.Transient overexpression in Nicotiana benthamiana leaves of SmMyb1 and of a C-terminal SmMyb1 truncated form (SmMyb1Δ9 resulted in anthocyanin accumulation only of SmMyb1 agro-infiltrated leaves. A yeast two-hybrid assay confirmed the interaction of both SmMyb1 and SmMyb1Δ9 with an anthocyanin-related potato bHLH1 TF. Interestingly, a doubled amount of CGA was detected in both SmMyb1 and SmMyb1Δ9 agro-infiltrated leaves, thus suggesting that the N-terminal region of SmMyb1 is sufficient to activate its synthesis. These data suggest that a deletion of

  2. Energy technology characterizations handbook: environmental pollution and control factors. Third edition

    Energy Technology Data Exchange (ETDEWEB)

    1983-03-01

    This Handbook deals with environmental characterization information for a range of energy-supply systems and provides supplementary information on environmental controls applicable to a select group of environmentally characterized energy systems. Environmental residuals, physical-resource requirements, and discussion of applicable standards are the principal information provided. The quantitative and qualitative data provided are useful for evaluating alternative policy and technical strategies and for assessing the environmental impact of facility siting, energy production, and environmental controls.

  3. Energy technology characterizations handbook: environmental pollution and control factors. Third edition

    International Nuclear Information System (INIS)

    1983-03-01

    This Handbook deals with environmental characterization information for a range of energy-supply systems and provides supplementary information on environmental controls applicable to a select group of environmentally characterized energy systems. Environmental residuals, physical-resource requirements, and discussion of applicable standards are the principal information provided. The quantitative and qualitative data provided are useful for evaluating alternative policy and technical strategies and for assessing the environmental impact of facility siting, energy production, and environmental controls

  4. In vitro characterization of high purity factor IX concentrates for the treatment of hemophilia B.

    Science.gov (United States)

    Limentani, S A; Gowell, K P; Deitcher, S R

    1995-04-01

    This study employed sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis and immunoblotting to assess the purity of seven high purity factor IX concentrates: Aimafix (Aima), AlphaNine-SD (Alpha Therapeutic), Factor IX VHP (Biotransfusion), Immunine (Immuno), Mononine (Armour Pharmaceutical), Nanotiv (Kabi Pharmacia), and 9MC (Blood Products Laboratory). The mean specific activity of these products ranged from 68 U factor IX/mg (Aimafix) to 246 U factor IX/mg (Mononine). SDS-PAGE analysis showed that the highest purity product, Mononine, had a single contaminating band under non-reducing conditions. Two additional bands were detected when this product was analyzed under reducing conditions. All other products had multiple contaminating bands that were more apparent under reducing than non-reducing conditions. The immunoblot for factor IX showed a dominant factor IX band for all products. In addition, visible light chain of factor IX was detected for AlphaNine-SD, Factor IX VHP, Immunine, Mononine, Nanotiv, and 9MC, suggesting that the factor IX in these products had undergone partial activation to factor IXa. Another contaminating band was visible at 49,500 for all of the products except 9MC. In addition to this band, high molecular weight contaminants were apparent for some products, most notably AlphaNine-SD. The identity of these bands is unknown. Immunoblotting failed to demonstrate factor VII as a contaminant of any of the high purity products, although factor VIIa could be detected in some lots of Immunine, Nanotiv, and 9MC by a clot-based assay. Factor X contaminated Aimafix, AlphaNine-SD, Factor IX VHP, Immunine, Nanotiv, and 9MC, but activation products of factor X were not detected.(ABSTRACT TRUNCATED AT 250 WORDS)

  5. Panel Data with Cross-Sectional Dependence Characterized by a Multi-Level Factor Structure

    DEFF Research Database (Denmark)

    Rodríguez-Caballero, Carlos Vladimir

    A panel data model with a multi-level cross-sectional dependence is proposed. The factor structure is driven by top-level common factors as well as non-pervasive factors. I propose a simple method to filter out the full factor structure that overcomes limitations in standard procedures which may...... mix up both levels of unobservable factors and may hamper the identification of the model. The model covers both stationary and non-stationary cases and takes into account other relevant features that make the model well suited to the analysis of many types of time series frequently addressed...

  6. Characterization of a Canine Tetranucleotide Microsatellite Marker Located in the First Intron of the Tumor Necrosis Factor Alpha Gene

    OpenAIRE

    WATANABE, Masashi; TANAKA, Kazuaki; TAKIZAWA, Tatsuya; SEGAWA, Kazuhito; NEO, Sakurako; TSUCHIYA, Ryo; MURATA, Michiko; MURAKAMI, Masaru; HISASUE, Masaharu

    2013-01-01

    ABSTRACT A polymorphic tetranucleotide (GAAT)n microsatellite in the first intron of the canine tumor necrosis factor alpha (TNFA) gene was characterized in this study; 139 dogs were analyzed: 22 Beagles, 26 Chihuahuas, 20 Miniature Dachshunds, 24 Miniature Poodles, 22 Pembroke Welsh Corgis and 25 Shiba Inus. We detected the presence of the 4 alleles (GAAT)5, (GAAT)6, (GAAT)7 and (GAAT)8, including 9 of the 10 expected genotypes. The expected heterozygosity (He) and the polymorphic informatio...

  7. Characterization of pancreatic transcription factor Pdx-1 binding sites using promoter microarray and serial analysis of chromatin occupancy

    OpenAIRE

    Keller, David M; McWeeney, Shannon; Arsenlis, Athanasios; Drouin, Jacques; Wright, Christopher V E; Wang, Haiyan; Wollheim, Claes; White, Peter; Kaestner, Klaus H; Goodman, Richard H

    2007-01-01

    The homeobox transcription factor Pdx-1 is necessary for pancreas organogenesis and beta cell function, however, most Pdx-1-regulated genes are unknown. To further the understanding of Pdx-1 in beta cell biology, we have characterized its genomic targets in NIT-1 cells, a mouse insulinoma cell line. To identify novel targets, we developed a microarray that includes traditional promoters as well as non-coding conserved elements, micro-RNAs, and elements identified through an unbiased approach ...

  8. Spondyloarthropaty characterization and factors determination of bad prognostic in a Colombian patients population

    International Nuclear Information System (INIS)

    Londono P, John Dario; Gonzalez, Luis Alonso; Ramirez, Luis Alberto; Santos, Pedro; Avila, Mabel; Santos, Ana Maria; Romero, Consuelo; Valle, Rafael Raul

    2005-01-01

    Purpose: to characterized a group of Colombian patients with diagnosis of Spondiloarthropaty (SpA) and establish the factors related with progression to Ankylosing Spondylitis (AS) in patients with recent diagnosis. Methods: a total of 139 consecutive (January 2000 to September 2004) patients with diagnosis of SpA (ESSG): 55 AS, 49 with undifferentiated SpA (u-Spa), 22 with Reactive Arthritis (ReA) and 13 with Psoriatic arthritis (PsA), were evaluated following the ASAS recommendations. The mean duration of disease was 8,5 +/- 9, 2 years. A familial history of SpA was registered in 18%: 25% AS, 11.5 u-SpA, 10% ReA, 25% PsA. Uveitis was noticed in 7.2%: 14.5% in AS and 4.2% in u-SpA. Clinical characteristics at onset were different among SpA subgroups. The Age of onset of disease (years) was 26.8 +/- 10 for all group and 22.4 +/- 7.2: AS, 27.8 +/- 8 u-SpA, 27.1 +/- 7.1 Are, 41.1:t 15.7 APs. Duration of arthritis at first episode o disease was 3.3 +/- 3.8 for all group and 4.2:t 5.2 for AS, 2.9 +/- 2.7 for u-SpA, 2.5 +/- 2.1 for Are and 3.4+/- 3.4 for APs. The Spinal Pain Duration at the first episode of disease was 5.8 +/- 6.3 for all group, 8.1 +/- 7.9q * for AS, 4.9+/-5 for u-SpA, 3.4+/- 2.6 for Are, 3.2+/- 2.3 for APs (q ANOVA P < 0.05* AS vs u-SpA, p < 0.025; :j: AS vs u-SpA p < 0.000; AS vs. total group p = 0.05). A total of 51 (29 u-SpA and 22 Are) patients began the illness after 2000 and they were followed by regular schedule: 11 patients progress to AS (5 u-SpA and 6 Are) in a period of 3.53+/-0.9 year. 10, 10 (77%) were HLA-B27 positive, the age of onset of the disease was 19.7+/-3.5 years, the duration of the first arthritis episode was 3,2+/-1,4 months, spinal pain 8,7+/-2.8 months, entesopathy 51+/-2.3 months. Our results indicate that demographic and clinical characteristic at onset (age, sex, duration of symptoms) may determine evolution over the time (prognosis) in patients with SpA

  9. Characterization of coagulation factor synthesis in nine human primary cell types

    NARCIS (Netherlands)

    Dashty, Monireh; Akbarkhanzadeh, Vishtaseb; Zeebregts, Clark J.; Spek, C. Arnold; Sijbrands, Eric J.; Peppelenbosch, Maikel P.; Rezaee, Farhad

    2012-01-01

    The coagulation/fibrinolysis system is essential for wound healing after vascular injury. According to the standard paradigm, the synthesis of most coagulation factors is restricted to liver, platelets and endothelium. We challenged this interpretation by measuring coagulation factors in nine human

  10. Characterization of senscence-associated NAC transcription factors in Barley (Hordeum Vulgare L.)

    DEFF Research Database (Denmark)

    Podzimska, Dagmara Agata

    , such as yield, biomass production and nutrient quality, and NAC (NAM, ATAF1/2 and CUC2) transcription factors are promising targets for the breeding. The aim of this thesis was thus to assess the role of NAC transcription factors in regulation of senescence in barley (Hordeum vulgare L.) and to contribute...

  11. Functional and Biochemical Characterization of Human Eukaryotic Translation Initiation Factor 3 in Living Cells

    Czech Academy of Sciences Publication Activity Database

    Wagner, Susan; Herrmannová, Anna; Malík, Radek; Peclinovská, L.; Valášek, Leoš Shivaya

    2014-01-01

    Roč. 34, č. 16 (2014), s. 3041-3052 ISSN 0270-7306 R&D Projects: GA ČR GAP305/10/0335 Institutional support: RVO:61388971 ; RVO:68378050 Keywords : MESSENGER-RNA RECRUITMENT * FACTOR EIF3 * IN-VIVO Subject RIV: CE - Biochemistry Impact factor: 4.777, year: 2014

  12. Characterizing Quality Factor of Niobium Resonators Using a Markov Chain Monte Carlo Approach

    Energy Technology Data Exchange (ETDEWEB)

    Basu Thakur, Ritoban; Tang, Qing Yang; McGeehan, Ryan; Carter, Faustin; Shirokoff, Erik

    2017-06-01

    The next generation of radiation detectors in high precision Cosmology, Astronomy, and particle-astrophysics experiments will rely heavily on superconducting microwave resonators and kinetic inductance devices. Understanding the physics of energy loss in these devices, in particular at low temperatures and powers, is vital. We present a comprehensive analysis framework, using Markov Chain Monte Carlo methods, to characterize loss due to two-level system in concert with quasi-particle dynamics in thin-film Nb resonators in the GHz range.

  13. Export of the Virulence Factors from Shigella Flexneri and Characterization of the mxi loci

    Science.gov (United States)

    1992-07-20

    studied in a monkey model for Shigella infection. The in vitro assays just described have been routinely used to closely examine each of the steps in...to Shigella protein antigens in Rhesus monkeys and humans infected with Shigella spp. Infect. Immun_ 53:57-63. Oaks, E. V. , and K. R. Turbyfill...thesis manuscript ent itled: ~Export of Virulence Faclors from Shigella f/exneri and characterization of the mxi loci. ~ beyond brief excerpts is

  14. Determination and use of scaling factors for waste characterization in nuclear power plants

    International Nuclear Information System (INIS)

    2009-01-01

    The management of radioactive waste requires knowledge of the amounts and concentrations of specific radionuclides in the waste. The particular radionuclides of interest or concern may vary according to the safety case or other requirements of the waste management facility and/or the regulations of the Member State. Many of the radionuclides important for long term management of the waste are difficult to measure (DTM) from the outside of a waste package. Identification and quantification of these DTM nuclides require methods that, in general, involve analysis of waste samples using complex radiochemical analysis to separate the various radionuclides for measurement. This method of direct measurement is generally not practical to employ for large numbers of waste packages or for many heterogeneous waste streams. An alternative approach that can be used in many cases is to exploit the relationship between some easy to measure (ETM) key nuclides, such as certain gamma emitters, and the DTM nuclides to derive information for the DTM nuclides of interest. The scaling factor (SF) method is such an approach that is widely used to evaluate these DTM nuclides. The SF method is based on developing a correlation between ETM and DTM nuclides. The activities of DTM nuclides in waste packages are then estimated by measuring the ETM nuclides based on gamma measurements from outside the package and applying the SFs to calculate the DTM activities. The international standard ISO 21238:2007 gives guidelines for the general methodology for empirically determining SFs to evaluate the radioactivity of DTM nuclides in low and intermediate level radioactive waste (LILW) packages. The objective of this report is to provide information on the international experience in the determination and use of SFs. This report shows actual examples of the ways in which SFs have been derived and applied in various Member States. Although the main focus of the ISO standard is on LILW from the operation

  15. Production of nodulation factors by Rhizobium meliloti: fermentation, purification and characterization of glycolipids.

    Science.gov (United States)

    Kohring, B; Baier, R; Niehaus, K; Pühler, A; Flaschel, E

    1997-12-01

    Lipooligosaccharides, synthesized by soil bacteria of the genera Rhizobium, are known to have multifunctional effects on a wide variety of plants as signal substances in symbiosis initiation, cell response elicitation and growth regulation. These so called nodulation (Nod-) factors represent interesting biotechnological products with respect to fundamental studies of symbiotic interactions as well as for potential applications. Therefore, a batch fermentation process on a scale of 30 l has been developed by means of the Rhizobium meliloti strain R.m. 1021 (pEK327) strongly overexpressing the genes for the synthesis of Nod factors. Induction by the flavone luteolin led to growth associated production of the lipooligosaccharides. Ultrafiltration was used for separating the biomass from the filtrate containing the extracellular Nod factors. Simultaneously, ultrafiltration reduced the amount of lipophilic substances, which would otherwise interfere with processes downstream. The second separation step consisted in adsorption on XAD-2, a nonspecific hydrophobic adsorptive resin. Adsorption of Nod factors was carried out by batch operation of a stirred tank. Desorption was performed by elution with methanol in a fixed bed column. A semi-preparative reversed phase HPLC (Polygoprep 100-30 C18) was chosen as the final purification step. The Nod factors were obtained after evaporation and lyophilization. Thus, about 600 mg of Nod factors were produced from 20 l of fermentation broth. The Nod factors produced by Rhizobium meliloti R.m. 1021 (pEK327) were identified by liquid secondary ion mass spectrometry and by reversed-phase HPLC as fluorescent derivatives of 2-aminobenzamide. The biological activity of the products was demonstrated by means of the root hair deformation (HAD-) assay.

  16. Synthesis and characterization of 18F-labeled active site inhibited factor VII (ASIS)

    DEFF Research Database (Denmark)

    Erlandsson, Maria; Nielsen, Carsten Haagen; Jeppesen, Troels Elmer

    2015-01-01

    Activated factor VII blocked in the active site with Phe-Phe-Arg-chloromethyl ketone (active site inhibited factor VII (ASIS)) is a 50-kDa protein that binds with high affinity to its receptor, tissue factor (TF). TF is a transmembrane glycoprotein that plays an important role in, for example......, thrombosis, metastasis, tumor growth, and tumor angiogenesis. The aim of this study was to develop an 18F-labeled ASIS derivative to assess TF expression in tumors. Active site inhibited factor VII was labeled using N-succinimidyl-4-[18F]fluorobenzoate, and the [18F]ASIS was purified on a PD-10 desalting...... column. The radiochemical yield was 25 ± 6%, the radiochemical purity was >97%, and the pseudospecific radioactivity was 35 ± 9 GBq/µmol. The binding efficacy was evaluated in pull-down experiments, which monitored the binding of unlabeled ASIS and [18F]ASIS to TF and to a specific anti-factor VII...

  17. Characterization of emission factors related to source activity for trichloroethylene degreasing and chrome plating processes.

    Science.gov (United States)

    Wadden, R A; Hawkins, J L; Scheff, P A; Franke, J E

    1991-09-01

    A study at an automotive parts fabrication plant evaluated four metal surface treatment processes during production conditions. The evaluation provides examples of how to estimate process emission factors from activity and air concentration data. The processes were open tank and enclosed tank degreasing with trichloroethylene (TCE), chromium conversion coating, and chromium electroplating. Area concentrations of TCE and chromium (Cr) were monitored for 1-hr periods at three distances from each process. Source activities at each process were recorded during each sampling interval. Emission rates were determined by applying appropriate mass balance models to the concentration patterns around each source. The emission factors obtained from regression analysis of the emission rate and activity data were 16.9 g TCE/basket of parts for the open-top degreaser; 1.0 g TCE/1000 parts for the enclosed degreaser; 1.48-1.64 mg Cr/1000 parts processed in the hot CrO3/HNO3 tank for the chrome conversion coating; and 5.35-9.17 mg Cr/rack of parts for chrome electroplating. The factors were also used to determine the efficiency of collection for the local exhaust systems serving each process. Although the number of observations were limited, these factors may be useful for providing initial estimates of emissions from similar processes in other settings.

  18. Characterizing Stressors and Modifiable Health Risk Factors among Homeless Smokers: An Exploratory Pilot Study

    Science.gov (United States)

    Kendzor, Darla E.; Reitzel, Lorraine R.; Businelle, Michael S.

    2015-01-01

    This pilot study was conducted to explore the associations between stressors related to homelessness and modifiable health risk factors (poor diet, insufficient physical activity, and overweight/obesity) and to provide direction for future research. Participants (N = 57) were homeless adults enrolled in a smoking cessation program. Analyses were…

  19. An OMERACT Initiative Toward Consensus to Identify and Characterize Candidate Contextual Factors

    DEFF Research Database (Denmark)

    Finger, Monika E; Boonen, Annelies; Woodworth, Thasia G

    2017-01-01

    OBJECTIVE: The importance of contextual factors (CF) for appropriate patient-specific care is widely acknowledged. However, evidence in clinical trials on how CF influence outcomes remains sparse. The 2014 Outcome Measures in Rheumatology (OMERACT) Handbook introduced the role of CF in outcome as...

  20. Synthesis and characterization of (18)F-labeled active site inhibited factor VII (ASIS).

    Science.gov (United States)

    Erlandsson, Maria; Nielsen, Carsten H; Jeppesen, Troels E; Kristensen, Jesper B; Petersen, Lars C; Madsen, Jacob; Kjaer, Andreas

    2015-05-15

    Activated factor VII blocked in the active site with Phe-Phe-Arg-chloromethyl ketone (active site inhibited factor VII (ASIS)) is a 50-kDa protein that binds with high affinity to its receptor, tissue factor (TF). TF is a transmembrane glycoprotein that plays an important role in, for example, thrombosis, metastasis, tumor growth, and tumor angiogenesis. The aim of this study was to develop an (18)F-labeled ASIS derivative to assess TF expression in tumors. Active site inhibited factor VII was labeled using N-succinimidyl-4-[(18)F]fluorobenzoate, and the [(18)F]ASIS was purified on a PD-10 desalting column. The radiochemical yield was 25 ± 6%, the radiochemical purity was >97%, and the pseudospecific radioactivity was 35 ± 9 GBq/µmol. The binding efficacy was evaluated in pull-down experiments, which monitored the binding of unlabeled ASIS and [(18)F]ASIS to TF and to a specific anti-factor VII antibody (F1A2-mAb). No significant difference in binding efficacy between [(18)F]ASIS and ASIS could be detected. Furthermore, [(18)F]ASIS was relatively stable in vitro and in vivo in mice. In conclusion, [(18)F]ASIS has for the first time been successfully synthesized as a possible positron emission tomography tracer to image TF expression levels. In vivo positron emission tomography studies to evaluate the full potential of [(18)F]ASIS are in progress. Copyright © 2015 John Wiley & Sons, Ltd.

  1. Mineralogy, occurrence of mineralization and temperature-pressure conditions of the Agh-Daragh polymetallic deposit in the Ahar-Arasbaran metallogenic area

    Directory of Open Access Journals (Sweden)

    Heydar Asgharzadeh Asl

    2017-07-01

    Full Text Available Introduction The Ahar-Arasbaran metallogenic area (Qara Dagh zone is located in the northwest of Iran and the western part of the Mazandaran Sea. At the base of the structural classification from Nabavi (1976, this area is situated in the Alborz-Azerbijan magmatic belt. Sheviar Dagh (Ahar batholite intrusive suite, subvolcanic and volcanic accompanied rocks with east-west trending and 30 km long serves as the main Eocene-Oligocene magmatic event in the north of the Ahar province. Considering geochemistry, this assemblage includes two shoshonitic and calc-alkaline to high-K calc-alkaline series which are the shoshonitic series in the central part and the calc-alkaline series outcrop in the western and eastern part (Aghazadeh, 2009. Textural characteristic, mineral chemistry and fluid inclusion studies were carried out at the tree part of the Agh-Daragh prospecting area. Material and methods A total of 50 samples were collected from the host rocks (including 4 trench and 250 m, ore deposit and altered rocks. Ten altered samples were analyzed for their mineral recognition by X-ray diffraction in the Iran Mineral Process and Research Center (IMPRC. Electron microprobe analyses (EMPA and backscattered electron (BSE images of minerals were obtained using a Cameca SX100 electron microprobe in the Iran Mineral Process and Research Center (IMPRC. An accelerating voltage of 15 to 25 kV and beam current of 20 mA was used for all analyses. Typical spot sizes ranged from 2 to 5 μm. A total of 5 double-polish thin sections representative of quartz samples were selected from mineralized veins after petrographic and field studies. Fluid inclusion microthermometry was conducted using a Linkam THMS600 heating–freezing stage (-190°C to +600°C mounted on a ZEISS Axioplan2 microscope in the fluid inclusion laboratory of the school of Earth Sciences of the Kharazmi University. The heating rate was 5–10 °C/min at higher temperatures (>100°C, with a

  2. Environmental, morphological, and productive characterization of Sardinian goats and use of latent explanatory factors for population analysis.

    Science.gov (United States)

    Vacca, G M; Paschino, P; Dettori, M L; Bergamaschi, M; Cipolat-Gotet, C; Bittante, G; Pazzola, M

    2016-09-01

    Dairy goat farming is practiced worldwide, within a range of different farming systems. Here we investigated the effects of environmental factors and morphology on milk traits of the Sardinian goat population. Sardinian goats are currently reared in Sardinia (Italy) in a low-input context, similar to many goat farming systems, especially in developing countries. Milk and morphological traits from 1,050 Sardinian goats from 42 farms were recorded. We observed a high variability regarding morphological traits, such as coat color, ear length and direction, horn presence, and udder shape. Such variability derived partly from the unplanned repeated crossbreeding of the native Sardinian goats with exotic breeds, especially Maltese goats. The farms located in the mountains were characterized by the traditional farming system and the lowest percentage of crossbred goats. Explanatory factors analysis was used to summarize the interrelated measured milk variables. The explanatory factor related to fat, protein, and energy content of milk (the "Quality" latent variable) explained about 30% of the variance of the whole data set of measured milk traits followed by the "Hygiene" (19%), "Production" (19%), and "Acidity" (11%) factors. The "Quality" and "Hygiene" factors were not affected by any of the farm classification items, whereas "Production" and "Acidity" were affected only by altitude and size of herds, respectively, indicating the adaptation of the local goat population to different environmental conditions. The use of latent explanatory factor analysis allowed us to clearly explain the large variability of milk traits, revealing that the Sardinian goat population cannot be divided into subpopulations based on milk attitude The factors, properly integrated with genetic data, may be useful tools in future selection programs.

  3. Candida Species From Eye Infections: Drug Susceptibility, Virulence Factors, and Molecular Characterization.

    Science.gov (United States)

    Ranjith, Konduri; Sontam, Bhavani; Sharma, Savitri; Joseph, Joveeta; Chathoth, Kanchana N; Sama, Kalyana C; Murthy, Somasheila I; Shivaji, Sisinthy

    2017-08-01

    To determine the type of Candida species in ocular infections and to investigate the relationship of antifungal susceptibility profile to virulence factors. Fifty isolates of yeast-like fungi from patients with keratitis, endophthalmitis, and orbital cellulitis were identified by Vitek-2 compact system and DNA sequencing of ITS1-5.8S-ITS2 regions of the rRNA gene, followed by phylogenetic analysis for phenotypic and genotypic identification, respectively. Minimum inhibitory concentration of six antifungal drugs was determined by E test/microbroth dilution methods. Phenotypic and genotypic methods were used to determine the virulence factors. Phylogenetic analysis showed the clustering of all isolates into eight distinct groups with a major cluster formed Candida parapsilosis (n = 21), which was the most common species by both Vitek 2 and DNA sequencing. Using χ2 test no significant difference was noted between the techniques except that Vitek 2 did not identify C. viswanathii, C. orthopsilosis, and two non-Candida genera. Of 43 tested Candida isolates high susceptibility to amphotericin B (39/43, 90.6%) and natamycin (43/43, 100%) was noted. While none of the isolates produced coagulase, all produced esterase and catalase. The potential to form biofilm was detected in 23/43 (53.4%) isolates. Distribution of virulence factors by heat map analysis showed difference in metabolic activity of biofilm producers from nonbiofilm producers. Identified by Vitek 2 and DNA sequencing methods C. parapsilosis was the most common species associated with eye infections. Irrespective of the virulence factors elaborated, the Candida isolates were susceptible to commonly used antifungal drugs such as amphotericin B and natamycin.

  4. Characterization of a receptor for human monocyte-derived neutrophil chemotactic factor/interleukin-8

    International Nuclear Information System (INIS)

    Grob, P.M.; David, E.; Warren, T.C.; DeLeon, R.P.; Farina, P.R.; Homon, C.A.

    1990-01-01

    Monocyte-derived neutrophil chemotactic factor/interleukin-8 (MDNCF/IL-8) is an 8,000-dalton protein produced by monocytes which exhibits activity as a chemoattractant for neutrophils with maximal activity achieved at a concentration of 50 ng/ml. This polypeptide has been iodinated by chloramine-T methodology (350 Ci/mM), and specific receptors for MDNCF/IL-8 have been detected on human neutrophils, U937 cells, THP-1 cells, and dimethyl sulfoxide-differentiated HL-60 cells. The binding of MDNCF/IL-8 to human neutrophils is not inhibited by interleukin-1 alpha, tumor necrosis factor-alpha, insulin, or epidermal growth factor. In addition, chemoattractants such as C5a, fMet-Leu-Phe, leukotriene B4, and platelet-activating factor fail to inhibit binding, suggesting that MDNCF/IL-8 utilizes a unique receptor. The receptor for MDNCF/IL-8 is apparently glycosylated since ligand binding is inhibited by the presence of wheat germ agglutinin, a lectin with a binding specificity for N-acetylglucosamine and neuraminic acid. Steady state binding experiments indicate Kd values of 4 and 0.5 nM and receptor numbers of 75,000 and 7,400 for human neutrophils and differentiated HL-60 cells, respectively. 125I-MDNCF/IL-8 bound to human neutrophils is rapidly internalized and subsequently released from cells as trichloroacetic acid-soluble radioactivity. Affinity labeling experiments suggest that the human neutrophil MDNCF/IL-8 receptor exhibits a mass of approximately 58,000 daltons

  5. Reactivity to Daily Stressors in Adulthood: The Importance of Stressor Type in Characterizing Risk Factors

    OpenAIRE

    Hay, Elizabeth L.; Diehl, Manfred

    2010-01-01

    This study examined daily stressors in adults aged 18 to 89 years (M = 49.6 years) over 30 days. We examined the role of individual factors (i.e., age, self-concept differentiation, perceived control) in physical and psychological reactivity to interpersonal, network, home, and health stressors. Findings were consistent with the perspective that adults were less reactive to stress on days they felt in control and that younger adults and adults with high self-concept differentiation (SCD) were...

  6. Study on Factors for Accurate Open Circuit Voltage Characterizations in Mn-Type Li-Ion Batteries

    Directory of Open Access Journals (Sweden)

    Natthawuth Somakettarin

    2017-03-01

    Full Text Available Open circuit voltage (OCV of lithium batteries has been of interest since the battery management system (BMS requires an accurate knowledge of the voltage characteristics of any Li-ion batteries. This article presents an OCV characteristic for lithium manganese oxide (LMO batteries under several experimental operating conditions, and discusses factors for accurate OCV determination. A test system is developed for OCV characterization based on the OCV pulse test method. Various factors for the OCV behavior, such as resting period, step-size of the pulse test, testing current amplitude, hysteresis phenomena, and terminal voltage relationship, are investigated and evaluated. To this end, a general OCV model based on state of charge (SOC tracking is developed and validated with satisfactory results.

  7. Partial Purification and Characterization of Anticoagulant Factor from the Snake (Echis Carinatus) Venom

    Science.gov (United States)

    Amrollahi Byoki, Elham; Zare Mirakabadi, Abbas

    2013-01-01

    Objective(s): Snake venoms contain complex mixture of proteins with biological activities. Some of these proteins affect blood coagulation and platelet function in different ways. Snake venom toxin may serve as a starting material for drug design to combat several pathophysiological problems such as cardiovascular disorders. In the present study, purification of anticoagulation factor from venom of snake (Echis carinatus) was studied. Materials and Methods: Anticoagulation activity of crude venom, fractions and purified peptide were determined by using prothrombin time (PT) and thrombin time (TT). Three fractions were partially purified from the venom of E. Carinatus by gel filtration on sephadex G-75 and final purification was performed by high-performance liquid chromatography (HPLC) with C18 column. A purified anticoagulant factor was derived which showed a single protein band in SDS-PAGE electrophoresis under reducing condition. Results: Results of PT and TT tests for purified peptide (EC217) were found to be 102±4.242 and < 5 min. respectively. Determination of molecular weight revealed that the active purified peptide (EC217) was about 30 KD. Conclusion: The present study showed that the venom of E. carinatus contains at least one anticoagulant factor. PMID:24494065

  8. Partial Purification and Characterization of Anticoagulant Factor from the Snake (Echis carinatus Venom

    Directory of Open Access Journals (Sweden)

    Elham Amrollahi Byoki

    2013-11-01

    Full Text Available   Objective(s: Snake venoms contain complex mixture of proteins with biological activities. Some of these proteins affect blood coagulation and platelet function in different ways. Snake venom toxin may serve as a starting material for drug design to combat several pathophysiological problems such as cardiovascular disorders. In the present study, purification of anticoagulation factor from venom of snake (Echis carinatus was studied. Anticoagulation activity of crude venom, fractions and purified peptide were determined by using prothrombin time (PT and thrombin time (TT. Three fractions were partially purified from the venom of E. Carinatus by gel filtration on sephadex G-75 and final purification was performed by high-performance liquid chromatography (HPLC with C18 column. A purified anticoagulant factor was derived which showed a single protein band in SDS-PAGE electrophoresis under reducing condition. Results of PT and TT tests for purified peptide (EC217 were found to be 102±4.242 and < 5 min. respectively. Determination of molecular weight revealed that the active purified peptide (EC217 was about 30 KD. In conclusion, the present study showed that the venom of E. carinatus contains at least one anticoagulant factor.

  9. Discovery and information-theoretic characterization of transcription factor binding sites that act cooperatively.

    Science.gov (United States)

    Clifford, Jacob; Adami, Christoph

    2015-09-02

    Transcription factor binding to the surface of DNA regulatory regions is one of the primary causes of regulating gene expression levels. A probabilistic approach to model protein-DNA interactions at the sequence level is through position weight matrices (PWMs) that estimate the joint probability of a DNA binding site sequence by assuming positional independence within the DNA sequence. Here we construct conditional PWMs that depend on the motif signatures in the flanking DNA sequence, by conditioning known binding site loci on the presence or absence of additional binding sites in the flanking sequence of each site's locus. Pooling known sites with similar flanking sequence patterns allows for the estimation of the conditional distribution function over the binding site sequences. We apply our model to the Dorsal transcription factor binding sites active in patterning the Dorsal-Ventral axis of Drosophila development. We find that those binding sites that cooperate with nearby Twist sites on average contain about 0.5 bits of information about the presence of Twist transcription factor binding sites in the flanking sequence. We also find that Dorsal binding site detectors conditioned on flanking sequence information make better predictions about what is a Dorsal site relative to background DNA than detection without information about flanking sequence features.

  10. HIV AND HCV COINFECTION: PREVALENCE, ASSOCIATED FACTORS AND GENOTYPE CHARACTERIZATION IN THE MIDWEST REGION OF BRAZIL

    Directory of Open Access Journals (Sweden)

    Solange Zacalusni Freitas

    2014-12-01

    Full Text Available A cross-sectional study on prevalence, associated factors and genotype distribution of HCV infection was conducted among 848 HIV-infected patients recruited at reference centers in the Midwest Region of Brazil. The prevalence rate of HIV-HCV coinfection was 6.9% (95% CI: 5.2 to 8.6. In multivariable analysis, increasing age, use of illicit drugs (injection and non-injection, a history of blood transfusion before 1994, and the absence of a steady partnership were significant independent associated factors for HIV-HCV coinfection. The phylogenetic analysis based on the NS5B region revealed the presence of two major circulating genotypes of HCV: genotypes 1 (58.3% and 3 (41.7%. The prevalence of HIV-HCV coinfection was lower than those reported in studies conducted with HIV-infected patients in different regions of Brazil, due to the fact that illicit drug use is not a frequent mode of HIV transmission in this region of Brazil. Serologic screening of HIV-patients for HCV before initiating antiretroviral treatment, a comprehensive identification of associated factors, and the implementation of effective harm reduction programs are highly recommended to provide useful information for treatment and to prevent HCV coinfection in these patients.

  11. HIV and HCV coinfection: prevalence, associated factors and genotype characterization in the Midwest Region of Brazil.

    Science.gov (United States)

    Freitas, Solange Zacalusni; Teles, Sheila Araújo; Lorenzo, Paulo Cesar; Puga, Marco Antonio Moreira; Tanaka, Tayana Serpa Ortiz; Thomaz, Danilo Yamamoto; Martins, Regina Maria Bringel; Druzian, Angelita Fernandes; Lindenberg, Andréa Siqueira Campos; Torres, Marina Sawada; Pereira, Sérgio A; Villar, Livia Melo; Lampe, Elisabete; Motta-Castro, Ana Rita Coimbra

    2014-01-01

    A cross-sectional study on prevalence, associated factors and genotype distribution of HCV infection was conducted among 848 HIV-infected patients recruited at reference centers in the Midwest Region of Brazil. The prevalence rate of HIV-HCV coinfection was 6.9% (95% CI: 5.2 to 8.6). In multivariable analysis, increasing age, use of illicit drugs (injection and non-injection), a history of blood transfusion before 1994, and the absence of a steady partnership were significant independent associated factors for HIV-HCV coinfection. The phylogenetic analysis based on the NS5B region revealed the presence of two major circulating genotypes of HCV: genotypes 1 (58.3%) and 3 (41.7%). The prevalence of HIV-HCV coinfection was lower than those reported in studies conducted with HIV-infected patients in different regions of Brazil, due to the fact that illicit drug use is not a frequent mode of HIV transmission in this region of Brazil. Serologic screening of HIV-patients for HCV before initiating antiretroviral treatment, a comprehensive identification of associated factors, and the implementation of effective harm reduction programs are highly recommended to provide useful information for treatment and to prevent HCV coinfection in these patients.

  12. Characterization of the interaction between the heavy and light chains of bovine factor Va.

    Science.gov (United States)

    Walker, F J

    1992-10-05

    Bovine factor Va has been previously been shown to consist of heavy (M(r) = 94,000) and light chains (M(r) = 81,000), that interact in a manner dependent upon the presence of either calcium or manganese ions. In an attempt to understand the mechanism of subunit interaction we have studied the effects of temperature and ions on factor Va stability. The rates of formation of factor Va from isolated chains and dissociation were temperature-dependent with an energy of activation of 6.2 and 1.3 kcal mol-1, respectively. The yield of factor Va from isolated chains was inversely related to the amount of time the chains were incubated at 4 degrees C. Incubation of individual chains revealed that the heavy chain is cold-labile, an effect that is reversible. Manganese ion was observed to prevent the conversion to the inactive form. High salt tends to stabilize the two-chain structure of factor Va, but is inhibitory to its formation from isolated chains. High concentrations of either manganese or calcium ions also inhibited reconstitution of activity. The light chain, in particular, was sensitive to the presence of manganese or calcium ion. Heavy chain that had been cleaved by activated protein C had a weakened interaction with the light chain, and the resulting complex had no procoagulant activity. Cooling of the heavy chain to 4 degrees C enhanced its intrinsic fluorescence. Manganese ion prevented some of this enhancement. The heavy chain fluorescence returned to the room temperature value with a half-life of approximately 10 min. In the presence of manganese ion relaxation was accelerated. The intrinsic fluorescence of activated protein C-cleaved heavy chain was not increased when the temperature was decreased. These data suggest that the heavy chain can exist in two forms. Elevated temperature converts it to a form that can bind ions and have a productive interaction with the light chain. However, conditions that prevent the heavy chain from combining with the light

  13. Development of characterization factors for metals in 7 EU water archetypes

    DEFF Research Database (Denmark)

    Dong, Yan; Hauschild, Michael Zwicky

    of the metals, WHAM 7.0 is used to model the metal speciation, Kd values and bioavailability, while the Free Ion Activity Model (FIAM) is used to model the ecotoxicity effect. The resulting archetype-specific CFs show up to ~4 orders of magnitude difference for Cr and Be. This indicates that the toxicity...... in the frequent situation where no information is available of the specific water type into which the metal emission occurs, site generic average factors are also calculated and different approaches to averaging across archetypes are investigated and discussed....

  14. Characterizing adult attention-deficit/hyperactivity-disorder and comorbid borderline personality disorder: ADHD symptoms, psychopathology, cognitive functioning and psychosocial factors.

    Science.gov (United States)

    O'Malley, G K; McHugh, L; Mac Giollabhui, N; Bramham, J

    2016-01-01

    To characterize adults with comorbid attention-deficit/hyperactivity-disorder (ADHD) and borderline personality disorder (BPD) with regard to ADHD symptoms, psychopathology, cognitive functioning and psychosocial factors. A between-group design compared a group of individuals diagnosed with ADHD (n=40) with a group diagnosed with BPD and who also met the criteria for ADHD (ADHD+BPD) (n=20). Significant differences were observed for both childhood and current impulsivity symptoms, whereby ADHD+BPD exhibited increased impulsivity; no differences on self-report and cognitive measures of impulsivity were reported. The ADHD+BPD group scored significantly higher on measures of depression, anxiety and numerous other axis I and II conditions. The ADHD+BPD group scored significantly lower on most measures of intellectual functioning and attention, however largely not on those relating to response inhibition. Furthermore, group differences were observed for psychosocial factors, including education, substance use and criminal record. Comorbid ADHD and BPD is characterized by more symptoms of impulsivity, additional psychopathology, comparatively lower intellectual and attentional functioning and increased psychosocial difficulties. Copyright © 2015. Published by Elsevier Masson SAS.

  15. The primary extinction and static Debye-Waller factor in the characterization of textured nickel by X-ray diffraction

    International Nuclear Information System (INIS)

    Kryshtab, T.; Palacios G, J.; Cadena A, A.; Kryvko, A.

    2015-01-01

    The texture analysis using X-ray diffraction (XRD) implies measurement of pole figures (Pf) from the diffracted intensities considering the model of kinematical dispersion. The extinction phenomenon results in a decrease of diffracted intensity and that in turn in a decrease of pole densities (Pds). The phenomenon appears in the kinematical theory of XRD as the primary extinction and the secondary extinction to characterize the loss of intensity of kinematical dispersion. In turn, the static Debye-Waller factor is an integral characteristic of defects in crystals that is introduced in the kinematical theory of XRD and also is used in dynamical theory of XRD. In this work the correlation between the primary extinction coefficient and the static Debye-Waller factor in the case of textured nickel was determined. The value of static Debye-Waller factor was determined from the value of the calculated primary extinction coefficient. For the evaluation there were used Pds in the maxima of Pf obtained for 111 and 200 reflections with Mo Kα radiation, and the Pds in the maxima of Pf obtained for the first and second orders of these reflections with Cu Kα and Co Kα radiations. There were calculated the dislocation densities in grains using values of static Debye-Waller factor and the extinction coefficients. The dislocation densities calculated from these two characteristics are practically equal. (Author)

  16. Falls in institutions for older adults: characterization of fall occurrences and associated risk factors

    Directory of Open Access Journals (Sweden)

    Cristina Rosa Soares Lavareda Baixinho

    2015-12-01

    Full Text Available Falls are the main accident for older adults, with consequences on functionality. Older adults impose restrictions or have restrictions imposed on their activities for fear of new falls. This prospective longitudinal study was conducted with 104 institutionalized older adults during six months with the following goals: to determine the prevalence of falls, to characterize the falls according to place, time, resulting injuries, supervision of the older adult, action performed at the time of the fall, and to relate the occurrence of the fall to the risk of falling, medical diagnoses, number of medications in use, type of medication, degree of dependency, age, and gender. The prevalence of falls was 37.5%, and they happened mostly in the bedroom, while walking after getting up from the bed. Those under risk in the Morse Fall Scale (p=0.034 and on sedatives (p=0.007 face a higher prevalence of falls. This study enables the possibility of making suggestions for practice, training and investigation.

  17. Genome-wide identification and characterization of GRAS transcription factors in tomato (Solanum lycopersicum).

    Science.gov (United States)

    Niu, Yiling; Zhao, Tingting; Xu, Xiangyang; Li, Jingfu

    2017-01-01

    Solanum lycopersicum , belonging to Solanaceae, is one of the commonly used model plants. The GRAS genes are transcriptional regulators, which play a significant role in plant growth and development, and the functions of several GRAS genes have been recognized, such as, axillary shoot meristem formation, radial root patterning, phytohormones (gibberellins) signal transduction, light signaling, and abiotic/biotic stress; however, only a few of these were identified and functionally characterized. In this study, a gene family was analyzed comprehensively with respect to phylogeny, gene structure, chromosomal localization, and expression pattern; the 54 GRAS members were screened from tomato by bioinformatics for the first time. The GRAS genes among tomato, Arabidopsis , rice, and grapevine were rebuilt to form a phylogenomic tree, which was divided into ten groups according to the previous classification of Arabidopsis and rice. A multiple sequence alignment exhibited the typical GRAS domain and conserved motifs similar to other gene families. Both the segmental and tandem duplications contributed significantly to the expansion and evolution of the GRAS gene family in tomato; the expression patterns across a variety of tissues and biotic conditions revealed potentially different functions of GRAS genes in tomato development and stress responses. Altogether, this study provides valuable information and robust candidate genes for future functional analysis for improving the resistance of tomato growth.

  18. Molecular cloning, characterization and expression analysis of coagulation factor VII gene in grass carp (Ctenopharyngodon idella).

    Science.gov (United States)

    Liu, Qiaolin; Xu, Baohong; Xiao, Tiaoyi; Su, Jianming; Zhong, Lei

    2013-08-01

    Coagulation factor VII has been studied in several species but, to date, not in grass carp (Ctenopharyngodon idella), a commercially important freshwater fish found in China. In this study, the full-length cDNA of grass carp coagulation factor VII (GcCFVII) was cloned using a RACE-Ready cDNA Kit, grass carp were challenged with a hemorrhagic virus, and temporal expression profiles of GcCFVII in the thymus, gills, liver, spleen, and head kidney were examined at 0 h, 24 h, 48 h, 72 h, 96 h, and 138 h using fluorescence quantitative PCR. The results showed the 1480 bp GcCFVII to contain three conservative motifs: Gla, EGF-CA, and Tryp-SPc, similar to other species. Phylogenetic analysis showed the evolution of GcCFVII gene to be consistent with the evolution of the species. After viral challenge, GcCFVII expression in five tissues of grass carp showed different patterns of fluctuation. These results provide a solid basis for further investigation of GcCFVII and its relationship with grass carp hemorrhage. Copyright © 2013 Elsevier Ltd. All rights reserved.

  19. Development and characterization of antiserum to murine granulocyte-macrophage colony-stimulating factor

    International Nuclear Information System (INIS)

    Mochizuki, D.Y.; Eisenman, J.R.; Conlon, P.J.; Park, L.S.; Urdal, D.L.

    1986-01-01

    The expression in yeast of a cDNA clone encoding murine granulocyte-macrophage colony-stimulating factor (GM-CSF) has made possible the purification of large quantities of this recombinant protein. Rabbits immunized with pure recombinant GM-CSF generated antibodies that were shown to be specific for both recombinant GM-CSF and GM-CSF isolated from natural sources. Other lymphokines, including IL 1β, IL 2, IL 3, and recombinant human GM-CSF did not react with the antiserum. The antiserum together with recombinant GM-CSF that had been radiolabeled with 125 I to high specific activity, formed the foundation for a rapid, sensitive, and quantitative radioimmunoassay specific for murine GM-CSF. Furthermore, the antiserum was found to inhibit the biologic activities of GM-CSF as measured in both a bone marrow proliferation assay and a colony assay, and thus should prove to be a useful reagent for dissecting the complex growth factor activities involved in murine hematopoiesis

  20. Isolation and characterization of StERF transcription factor genes from potato (Solanum tuberosum L.).

    Science.gov (United States)

    Wang, Zemin; Zhang, Ning; Zhou, Xiangyan; Fan, Qiang; Si, Huaijun; Wang, Di

    2015-04-01

    Ethylene response factor (ERF) is a major subfamily of the AP2/ERF family and plays significant roles in the regulation of abiotic- and biotic-stress responses. ERF proteins can interact with the GCC-box cis-element and then initiate a transcriptional cascade activating downstream ethylene response and enhancing plant stress tolerance. In this research, we cloned five StERF genes from potato (Solanum tuberosum L.). The expressional analysis of StERF genes revealed that they showed tissue- or organ-specific expression patterns and the expression levels in leaf, stem, root, flower, and tuber were different. The assays of quantitative real-time polymerase chain reaction (qRT-PCR) and the reverse transcription-PCR (RT-PCR) showed that the expression of five StERF genes was regulated by ethephon, methyl jasmonate (MeJA), salt and drought stress. The result from the yeast one-hybrid experiment showed that five StERFs had trans-activation activity and could specifically bind to the GCC-box cis-elements. The StERFs responded to abiotic factors and hormones suggested that they possibly had diverse roles in stress and hormone regulation of potato. Copyright © 2015 Académie des sciences. Published by Elsevier SAS. All rights reserved.

  1. Characterization and risk factors for pressure ulcers in the hospitalized elderly

    Directory of Open Access Journals (Sweden)

    Chrystiany Plácido de Brito Vieira

    2014-10-01

    Full Text Available This study aimed to identify the socio-demographic and clinical characteristics; obtain the risk factors for pressure ulcers identified in hospitalized senior citizens. This is a prospective, descriptive, and quantitative study. The sample consisted of 215 hospitalized elderly people in a public hospital in Teresina, PI, Brazil, in orthopedic, neurological, medical, and surgical clinics, and Intensive Care Units from November/2012 to February/2013. Data collection happened through interviews, physical examinations, and medical records. The average age was 71.8 years old, most patients were female (55.3%, with low education, and married. The majority were taken into orthopedics (42.3%. For the length of hospital stay, the average was of 12.4 days. The most prevalent risk factors were hypertension, use of analgesics, antihypertensives, and anticoagulants. Eighty-seven senior citizens (40.5% presented low risk. One highlights the importance of nursing care in the prevention and assessment of risk for pressure ulcers in order to reduce its incidence in hospitals.

  2. Characterization of clinical and genetic risk factors associated with dyslipidemia after kidney transplantation.

    Science.gov (United States)

    Numakura, Kazuyuki; Kagaya, Hideaki; Yamamoto, Ryohei; Komine, Naoki; Saito, Mitsuru; Hiroshi, Tsuruta; Akihama, Susumu; Inoue, Takamitsu; Narita, Shintaro; Tsuchiya, Norihiko; Habuchi, Tomonori; Niioka, Takenori; Miura, Masatomo; Satoh, Shigeru

    2015-01-01

    We determined the prevalence of dyslipidemia in a Japanese cohort of renal allograft recipients and investigated clinical and genetic characteristics associated with having the disease. In total, 126 patients that received renal allograft transplants between February 2002 and August 2011 were studied, of which 44 recipients (34.9%) were diagnosed with dyslipidemia at 1 year after transplantation. Three clinical factors were associated with a risk of having dyslipidemia: a higher prevalence of disease observed among female than male patients (P = 0.021) and treatment with high mycophenolate mofetil (P = 0.012) and prednisolone (P = 0.023) doses per body weight at 28 days after transplantation. The genetic association between dyslipidemia and 60 previously described genetic polymorphisms in 38 putative disease-associated genes was analyzed. The frequency of dyslipidemia was significantly higher in patients with the glucocorticoid receptor (NR3C1) Bcl1 G allele than in those with the CC genotype (P = 0.001). A multivariate analysis revealed that the NR3C1 Bcl1 G allele was a significant risk factor for the prevalence of dyslipidemia (odds ratio = 4.6; 95% confidence interval = 1.8-12.2). These findings may aid in predicting a patient's risk of developing dyslipidemia.

  3. Characterization of Clinical and Genetic Risk Factors Associated with Dyslipidemia after Kidney Transplantation

    Science.gov (United States)

    Numakura, Kazuyuki; Kagaya, Hideaki; Yamamoto, Ryohei; Komine, Naoki; Saito, Mitsuru; Hiroshi, Tsuruta; Akihama, Susumu; Narita, Shintaro; Tsuchiya, Norihiko; Habuchi, Tomonori; Niioka, Takenori; Miura, Masatomo; Satoh, Shigeru

    2015-01-01

    We determined the prevalence of dyslipidemia in a Japanese cohort of renal allograft recipients and investigated clinical and genetic characteristics associated with having the disease. In total, 126 patients that received renal allograft transplants between February 2002 and August 2011 were studied, of which 44 recipients (34.9%) were diagnosed with dyslipidemia at 1 year after transplantation. Three clinical factors were associated with a risk of having dyslipidemia: a higher prevalence of disease observed among female than male patients (P = 0.021) and treatment with high mycophenolate mofetil (P = 0.012) and prednisolone (P = 0.023) doses per body weight at 28 days after transplantation. The genetic association between dyslipidemia and 60 previously described genetic polymorphisms in 38 putative disease-associated genes was analyzed. The frequency of dyslipidemia was significantly higher in patients with the glucocorticoid receptor (NR3C1) Bcl1 G allele than in those with the CC genotype (P = 0.001). A multivariate analysis revealed that the NR3C1 Bcl1 G allele was a significant risk factor for the prevalence of dyslipidemia (odds ratio = 4.6; 95% confidence interval = 1.8–12.2). These findings may aid in predicting a patient's risk of developing dyslipidemia. PMID:25944971

  4. Multiple Stage Ore Formation in the Chadormalu Iron Deposit, Bafq Metallogenic Province, Central Iran: Evidence from BSE Imaging and Apatite EPMA and LA-ICP-MS U-Pb Geochronology

    Directory of Open Access Journals (Sweden)

    Hassan Heidarian

    2018-02-01

    Full Text Available The Chadormalu magnetite-apatite deposit in Bafq metallogenic province, Central Iran, is hosted in the late Precambrian-lower Cambrian volcano-sedimentary rocks with sodic, calcic, and potassic alterations characteristic of iron oxide copper-gold (IOCG and iron oxide-apatite (IOA ore systems. Apatite occurs as scattered irregular veinlets and disseminated grains, respectively, within and in the marginal parts of the main ore-body, as well as apatite-magnetite veins in altered wall rocks. Textural evidence (SEM-BSE images of these apatites shows primary bright, and secondary dark areas with inclusions of monazite/xenotime. The primary, monazite-free fluorapatite contains higher concentrations of Na, Si, S, and light rare earth elements (LREE. The apatite was altered by hydrothermal events that led to leaching of Na, Si, and REE + Y, and development of the dark apatite. The bright apatite yielded two U-Pb age populations, an older dominant age of 490 ± 21 Ma, similar to other iron deposits in the Bafq district and associated intrusions, and a younger age of 246 ± 17 Ma. The dark apatite yielded a U-Pb age of 437 ± 12 Ma. Our data suggest that hydrothermal magmatic fluids contributed to formation of the primary fluorapatite, and sodic and calcic alterations. The primary apatite reequilibrated with basinal brines in at least two regional extensions and basin developments in Silurian and Triassic in Central Iran.

  5. Immunohistochemical characterization and evaluation of prognostic factors in canine oral melanomas with osteocartilaginous differentiation.

    Science.gov (United States)

    Sánchez, J; Ramirez, G A; Buendia, A J; Vilafranca, M; Martinez, C M; Altimira, J; Navarro, J A

    2007-09-01

    Melanomas are the most common malignant oral neoplasm in dogs. Osteocartilaginous differentiation in oral melanomas is a rare feature described both in veterinary and human medicine. Here, 10 cases of this type of neoplasm were used to study their immunohistochemical, biological, and clinical characteristics. Reactivity for S100 and melan A antigen was evaluated, and 4 prognosis factors (mitotic index, invasiveness of epithelium, nuclear atypia, and proliferation index) were analyzed and correlated with the clinical course of the neoplasms after diagnosis. Immunohistochemical analysis of the studied neoplasms, including the osteocartilaginous areas, showed positive immunoreaction for S100 and melan A, except in one dog, which was negative for melan A. Analysis of the results showed that oral melamonas with osteocartilaginous differentiation have a clinical course similar to that of other melanomas in the oral cavity. Analysis of the mitotic index and the expression of proliferation marker Ki-67 could be useful tools for predicting the biological behavior of these neoplasms.

  6. Characterization of a "TRAMP-like" co-factor of the human RNA exosome

    DEFF Research Database (Denmark)

    Christensen, Marianne Skovgaard; Kristiansen, Maiken Søndergaard; Lubas, Michal Szymon

    Genome-wide studies in yeast, plants and humans have revealed numerous new transcripts in what was previously thought to be silent DNA or junk DNA. One class of non-coding transcript discovered recently is the PROMoter uPstream Transcripts (PROMPTs), which is only seen upon depletion of the RNA...... exosome, the major 3’-5’ exonuclease complex in human cells. PROMPTs have a lot in common with the yeast Cryptic Unstable Transcripts (CUTs), which are degraded by the concerted effort of the exosome, and its co-factor complex TRAMP (Trf4p/Air1p/Mtr4p). We have identified human proteins with functional...... similarities to components of the yeast TRAMP complex, and show that these are involved in the degradation of PROMPTs. While, these proteins form transient complexes with the exosome, our preliminary results also indicate that complex formation can occur directly with catalytic components of the exosome...

  7. Characterization of water-forming NADH oxidases for co-factor regeneration

    DEFF Research Database (Denmark)

    Rehn, Gustav; Pedersen, Asbjørn Toftgaard; J. Charnock, Simon

    an environmentaland economic perspective [1]. Alcohol dehydrogenases (ADH) offer one such alternative. However, the reaction requires the oxidized nicotinamide co-factor (NAD+) that must be recycled due to its high cost contribution. One regeneration method that offers certain advantages is the oxidation of NADH......Traditional chemical methods for alcohol oxidation are often associated with issues such as high consumption of expensive oxidizing agents, generation of metal waste and the use of environmentally undesirable organic solvents. Developing green, selective catalysts is therefore important from...... using water forming NADH oxidases (NOX-2). The implementation of the ADH/NOX system for alcohol oxidation, however, requires consideration of several different issues. Enzyme activity and stability at relevant pH and temperature conditions, but also the tolerance to the substrates and products present...

  8. The glycosylation and characterization of the candidate Gc macrophage activating factor

    DEFF Research Database (Denmark)

    Ravnsborg, Tina; Olsen, Dorthe T; Thysen, Anna Hammerich

    2010-01-01

    The vitamin D binding protein, Gc globulin, has in recent years received some attention for its role as precursor for the extremely potent macrophage activating factor (GcMAF). An O-linked trisaccharide has been allocated to the threonine residue at position 420 in two of the three most common...... isoforms of Gc globulin (Gc1s and Gc1f). A substitution for a lysine residue at position 420 in Gc2 prevents this isoform from being glycosylated at that position. It has been suggested that Gc globulin subjected sequentially to sialidase and galactosidase treatment generates GcMAF in the form of Gc...... globulin with only a single GalNAc attached to T420. In this study we confirm the location of a linear trisaccharide on T420. Furthermore, we provide the first structural evidence of the generation of the proposed GcMAF by use of glycosidase treatment and mass spectrometry. Additionally the generated GcMAF...

  9. Isolation, purification and partial characterization of early pregnancy factor (EPF) from sera of pregnant women.

    Science.gov (United States)

    Haq, A; Mothi, B A; Al-Hussein, K; Al-Tufail, M; Hollanders, J; Jaroudi, K; Al-Waili, N; Shabani, M

    2001-05-29

    Early pregnancy factor (EPF) is a pregnancy protein, which is secreted into the maternal serum 12-16 hours after fertilization. It is thought to be an immunosuppressive molecule. EPF is detected in pregnant woman's serum by the rosette inhibition assay (RIA). In this study, EPF was purified from the pregnant woman's sera by using ion exchange chromatography and analyzed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The proteins which showed a positive result with the RIA, were found to be 35 kDa and 17 kDa molecular weights. The biological activities of these proteins were stable upon heat treatment at 56 degrees C for 30 min. Proteins isolated and purified in this study might be of great significance to the field of human reproduction with particular reference to pregnancy and recurrent abortion.

  10. A Genome-Scale Resource for the Functional Characterization of Arabidopsis Transcription Factors

    Directory of Open Access Journals (Sweden)

    Jose L. Pruneda-Paz

    2014-07-01

    Full Text Available Extensive transcriptional networks play major roles in cellular and organismal functions. Transcript levels are in part determined by the combinatorial and overlapping functions of multiple transcription factors (TFs bound to gene promoters. Thus, TF-promoter interactions provide the basic molecular wiring of transcriptional regulatory networks. In plants, discovery of the functional roles of TFs is limited by an increased complexity of network circuitry due to a significant expansion of TF families. Here, we present the construction of a comprehensive collection of Arabidopsis TFs clones created to provide a versatile resource for uncovering TF biological functions. We leveraged this collection by implementing a high-throughput DNA binding assay and identified direct regulators of a key clock gene (CCA1 that provide molecular links between different signaling modules and the circadian clock. The resources introduced in this work will significantly contribute to a better understanding of the transcriptional regulatory landscape of plant genomes.

  11. Characterizing the astrophysical S factor for 12C+12C fusion with wave-packet dynamics

    Science.gov (United States)

    Diaz-Torres, Alexis; Wiescher, Michael

    2018-05-01

    A quantitative study of the astrophysically important subbarrier fusion of 12C+12C is presented. Low-energy collisions are described in the body-fixed reference frame using wave-packet dynamics within a nuclear molecular picture. A collective Hamiltonian drives the time propagation of the wave packet through the collective potential-energy landscape. The fusion imaginary potential for specific dinuclear configurations is crucial for understanding the appearance of resonances in the fusion cross section. The theoretical subbarrier fusion cross sections explain some observed resonant structures in the astrophysical S factor. These cross sections monotonically decline towards stellar energies. The structures in the data that are not explained are possibly due to cluster effects in the nuclear molecule, which need to be included in the present approach.

  12. Characterization and localization of metal-responsive-element-binding transcription factors from tilapia

    International Nuclear Information System (INIS)

    Cheung, Andrew Pok-Lap; Au, Candy Yee-Man; Chan, William Wai-Lun; Chan, King Ming

    2010-01-01

    Two isoforms of MTF-1, MTF-1L (long form) and MTF-1S (short form), were cloned in tilapia (Ti) and characterized in a tilapia liver cell line, Hepa-T1. The cloned tiMTF-1L has the characteristics of all of the tiMTF-1S identified so far with the zinc finger domain having six fingers, the acidic-rich, proline-rich, and serine/threonine-rich domains; however, the short form encodes for the zinc finger domain with five zinc fingers only and no other domains. The transient transfection of tiMTF-1L into human HepG2 cells showed both constitutive and zinc-induced metal-responsive-element (MRE)-driven reporter gene expression. However, the transfection of tiMTF-1S (which lacks all three transactivation domains) into a human cell line showed reduced transcriptional activities compared with an endogenous control in both basal- and Zn 2+ -induced conditions. The tiMTF-1 isoforms were tagged with GFP and transfected into Hepa-T1 cells (tilapia hepatocytes). The nuclear translocation of tiMTF-1L was observed when the cells were exposed to a sufficient concentration of metals for 6 h. However, tiMTF-1S, was localized in the nucleus with or without metal treatment. Electrophoretic mobility shift assay (EMSA) confirmed that both of the isoforms were able to bind to the MRE specifically in vitro. Tissue distribution studies showed that tiMTF-1L was more abundant than tiMTF-1S in all of the tissues tested.

  13. Characterization and localization of metal-responsive-element-binding transcription factors from tilapia

    Energy Technology Data Exchange (ETDEWEB)

    Cheung, Andrew Pok-Lap; Au, Candy Yee-Man; Chan, William Wai-Lun [Department of Biochemistry, Chinese University of Hong Kong, Sha Tin, N.T., Hong Kong (Hong Kong); Chan, King Ming, E-mail: kingchan@cuhk.edu.hk [Department of Biochemistry, Chinese University of Hong Kong, Sha Tin, N.T., Hong Kong (Hong Kong)

    2010-08-01

    Two isoforms of MTF-1, MTF-1L (long form) and MTF-1S (short form), were cloned in tilapia (Ti) and characterized in a tilapia liver cell line, Hepa-T1. The cloned tiMTF-1L has the characteristics of all of the tiMTF-1S identified so far with the zinc finger domain having six fingers, the acidic-rich, proline-rich, and serine/threonine-rich domains; however, the short form encodes for the zinc finger domain with five zinc fingers only and no other domains. The transient transfection of tiMTF-1L into human HepG2 cells showed both constitutive and zinc-induced metal-responsive-element (MRE)-driven reporter gene expression. However, the transfection of tiMTF-1S (which lacks all three transactivation domains) into a human cell line showed reduced transcriptional activities compared with an endogenous control in both basal- and Zn{sup 2+}-induced conditions. The tiMTF-1 isoforms were tagged with GFP and transfected into Hepa-T1 cells (tilapia hepatocytes). The nuclear translocation of tiMTF-1L was observed when the cells were exposed to a sufficient concentration of metals for 6 h. However, tiMTF-1S, was localized in the nucleus with or without metal treatment. Electrophoretic mobility shift assay (EMSA) confirmed that both of the isoforms were able to bind to the MRE specifically in vitro. Tissue distribution studies showed that tiMTF-1L was more abundant than tiMTF-1S in all of the tissues tested.

  14. Molecular Cloning and Characterization of Growth Factor Receptor Bound-Protein in Clonorchis sinensis

    Science.gov (United States)

    Bai, Xuelian; Lee, Ji-Yun; Kim, Tae Im; Dai, Fuhong; Lee, Tae-Jin; Hong, Sung-Jong

    2014-01-01

    Background Clonorchis sinensis causes clonorchiasis, a potentially serious disease. Growth factor receptor-bound protein 2 (Grb2) is a cytosolic protein conserved among animals and plays roles in cellular functions such as meiosis, organogenesis and energy metabolism. In the present study, we report first molecular characters of growth factor receptor bound-protein (CsGrb2) from C. sinensis as counter part of Grb2 from animals and its possible functions in development and organogenesis of C. sinensis. Methodology/Principal Findings A CsGrb2 cDNA clone retrieved from the C. sinensis transcriptome encoded a polypeptide with a SH3-SH2-SH3 structure. Recombinant CsGrb2 was bacterially produced and purified to homogeneity. Native CsGrb2 with estimated molecular weight was identified from C. sinensis adult extract by western blotting using a mouse immune serum to recombinant CsGrb2. CsGrb2 transcripts was more abundant in the metacercariae than in the adults. Immunohistochemical staining showed that CsGrb2 was localized to the suckers, mesenchymal tissues, sperms in seminal receptacle and ovary in the adults, and abundantly expressed in most organs of the metacercariae. Recombinant CsGrb2 was evaluated to be little useful as a serodiagnostic reagent for C. sinesis human infections. Conclusion Grb2 protein found in C. sinensis was conserved among animals and suggested to play a role in the organogenesis, energy metabolism and mitotic spermatogenesis of C. sinensis. These findings from C. sinensis provide wider understanding on diverse function of Grb2 in lower animals such as platyhelminths. PMID:24454892

  15. Molecular cloning and characterization of growth factor receptor bound-protein in Clonorchis sinensis.

    Directory of Open Access Journals (Sweden)

    Xuelian Bai

    Full Text Available BACKGROUND: Clonorchis sinensis causes clonorchiasis, a potentially serious disease. Growth factor receptor-bound protein 2 (Grb2 is a cytosolic protein conserved among animals and plays roles in cellular functions such as meiosis, organogenesis and energy metabolism. In the present study, we report first molecular characters of growth factor receptor bound-protein (CsGrb2 from C. sinensis as counter part of Grb2 from animals and its possible functions in development and organogenesis of C. sinensis. METHODOLOGY/PRINCIPAL FINDINGS: A CsGrb2 cDNA clone retrieved from the C. sinensis transcriptome encoded a polypeptide with a SH3-SH2-SH3 structure. Recombinant CsGrb2 was bacterially produced and purified to homogeneity. Native CsGrb2 with estimated molecular weight was identified from C. sinensis adult extract by western blotting using a mouse immune serum to recombinant CsGrb2. CsGrb2 transcripts was more abundant in the metacercariae than in the adults. Immunohistochemical staining showed that CsGrb2 was localized to the suckers, mesenchymal tissues, sperms in seminal receptacle and ovary in the adults, and abundantly expressed in most organs of the metacercariae. Recombinant CsGrb2 was evaluated to be little useful as a serodiagnostic reagent for C. sinesis human infections. CONCLUSION: Grb2 protein found in C. sinensis was conserved among animals and suggested to play a role in the organogenesis, energy metabolism and mitotic spermatogenesis of C. sinensis. These findings from C. sinensis provide wider understanding on diverse function of Grb2 in lower animals such as platyhelminths.

  16. Functional characterization of the copper transcription factor AfMac1 from Aspergillus fumigatus.

    Science.gov (United States)

    Park, Yong-Sung; Kim, Tae-Hyoung; Yun, Cheol-Won

    2017-07-03

    Although copper functions as a cofactor in many physiological processes, copper overload leads to harmful effects in living cells. Thus, copper homeostasis is tightly regulated. However, detailed copper metabolic pathways have not yet been identified in filamentous fungi. In this report, we investigated the copper transcription factor AfMac1 ( A spergillus f umigatus Mac1 homolog) and identified its regulatory mechanism in A. fumigatus AfMac1 has domains homologous to the DNA-binding and copper-binding domains of Mac1 from Saccharomyces cerevisiae , and AfMac1 efficiently complemented Mac1 in S. cerevisiae Expression of Afmac1 resulted in CTR1 up-regulation, and mutation of the DNA-binding domain of Afmac1 failed to activate CTR1 expression in S. cerevisiae The Afmac1 deletion strain of A. fumigatus failed to grow in copper-limited media, and its growth was restored by introducing ctrC We found that AfMac1 specifically bound to the promoter region of ctrC based on EMSA. The AfMac1-binding motif 5'-TGTGCTCA-3' was identified from the promoter region of ctrC , and the addition of mutant ctrC lacking the AfMac1-binding motif failed to up-regulate ctrC in A. fumigatus Furthermore, deletion of Afmac1 significantly reduced strain virulence and activated conidial killing activity by neutrophils and macrophages. Taken together, these results suggest that AfMac1 is a copper transcription factor that regulates cellular copper homeostasis in A. fumigatus . © 2017 The Author(s); published by Portland Press Limited on behalf of the Biochemical Society.

  17. Molecular characterization of two sub-family specific monoclonal antibodies to meningococcal Factor H binding protein

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    C. Lo Passo

    2018-04-01

    Full Text Available Factor H binding protein (FHbp is a component of two licensed vaccines for prevention of sepsis and meningitis caused by serogroup B meningococci. FHbp binds human Factor H (FH, which contributes to evasion of host immunity and FHbp sequence variants can be classified into two sub-families. Antibodies against FHbp elicit complement-mediated killing and can inhibit recruitment of FH to the bacterial surface. We report epitope mapping studies of two murine IgG mAbs, designated JAR 31 and JAR 36, isolated from a mouse immunized with FHbp in sub-family A, which is present in ∼30–40% of invasive isolates. In the present study, we tested the reactivity of mAbs JAR 31 and JAR 36 with seven natural FHbp sequence variants from different phylogenic groups. We screened bacteriophage-displayed peptide libraries to identify amino acid residues contributing to the JAR 36 epitope. Based on the reactivities of mAbs JAR 31 and JAR 36 with the seven FHbp variants, and the frequent occurrences of aspartate (D and lysine (K residues in the JAR 36-bound phage peptides, we selected six residues in the carboxyl-terminal region of FHbp for replacement with alanine (A. The D201A and K203A substitutions respectively eliminated and decreased binding of mAbs JAR 31 and JAR 36 to FHbp. These substitutions did not affect binding of the control mAb JAR 33 or of human FH. JAR 31 or JAR 36 mediated cooperative complement-mediated bactericidal activity with other anti-FHbp mAbs. The identification of two amino acid residues involved in the epitopes recognized by these anti-FHbp mAbs may contribute to a more complete understanding of the spatial requirements for cooperative anti-FHbp mAb bactericidal activity. Keywords: Biochemistry, Immunology, Microbiology, Molecular biology

  18. Identification and Characterization of msf, a Novel Virulence Factor in Haemophilus influenzae.

    Directory of Open Access Journals (Sweden)

    Jennifer M Kress-Bennett

    Full Text Available Haemophilus influenzae is an opportunistic pathogen. The emergence of virulent, non-typeable strains (NTHi emphasizes the importance of developing new interventional targets. We screened the NTHi supragenome for genes encoding surface-exposed proteins suggestive of immune evasion, identifying a large family containing Sel1-like repeats (SLRs. Clustering identified ten SLR-containing gene subfamilies, each with various numbers of SLRs per gene. Individual strains also had varying numbers of SLR-containing genes from one or more of the subfamilies. Statistical genetic analyses of gene possession among 210 NTHi strains typed as either disease or carriage found a significant association between possession of the SlrVA subfamily (which we have termed, macrophage survival factor, msf and the disease isolates. The PittII strain contains four chromosomally contiguous msf genes. Deleting all four of these genes (msfA1-4 (KO resulted in a highly significant decrease in phagocytosis and survival in macrophages; which was fully complemented by a single copy of the msfA1 gene. Using the chinchilla model of otitis media and invasive disease, the KO strain displayed a significant decrease in fitness compared to the WT in co-infections; and in single infections, the KO lost its ability to invade the brain. The singly complemented strain showed only a partial ability to compete with the WT suggesting gene dosage is important in vivo. The transcriptional profiles of the KO and WT in planktonic growth were compared using the NTHi supragenome array, which revealed highly significant changes in the expression of operons involved in virulence and anaerobiosis. These findings demonstrate that the msfA1-4 genes are virulence factors for phagocytosis, persistence, and trafficking to non-mucosal sites.

  19. Identification and characterization of a novel nanobody against human placental growth factor to modulate angiogenesis.

    Science.gov (United States)

    Arezumand, Roghaye; Mahdian, Reza; Zeinali, Sirous; Hassanzadeh-Ghassabeh, Gholamreza; Mansouri, Kamran; Khanahmad, Hossein; Namvar-Asl, Nabiollah; Rahimi, Hamzeh; Behdani, Mahdi; Cohan, Reza Ahangari; Eavazalipour, Mehdi; Ramazani, Ali; Muyldermans, Serge

    2016-10-01

    Placental growth factor (PlGF), a member of vascular endothelial growth factors (VEGF) family, is considered as an important antigen associated with pathological conditions such as cancer cell growth, and metastasis. PlGF-targeting via nanobody (Nb) therefore could be beneficial to modulate these pathologies. In this work, phage-display and computational approach was employed to develop a high affinity PlGF-specific Nb. An Nb library was constructed against human recombinant PlGF (rPlGF). After panning on immobilized rPlGF the periplasmic-extract (PE) of individual colonies were screened by ELISA (PE-ELISA). The 3D structures of selected Nbs were then homology modeled and energy minimized using the AMBER force field. Binding score calculations were also assessed to reveal possible Nb-PlGF interactions. Via ELISA-based affinity/specificity determinations, the best-qualified Nb was further evaluated by proliferation, migration, 3D capillary formation, invasion assays and on Chick chorioallantoic membrane (CAM) model. An immune library of 1.5×10 7 individual Nb clones was constructed. By PE-ELISA 12 clones with strong signals were selected. Three out of 12 sequenced Nbs (Nb-C13, Nb-C18 and Nb-C62) showed high binding scores ranging between -378.7 and -461kcal/mol. Compared to a control Nb, Nb-C18 significantly inhibited proliferation, migration and the 3D-capillary formation of HUVEC cells (p<0.05) with an EC 50 of 35nM, 42nM and 24nM and invasion of MDA-MB231was significantly suppressed (p<0.05) with an EC 50 of57nM. The result of the CAM assay shows that Nb-C18 could inhibit the vascular formation in the chicken chorioallantoic membrane. This Nb can be used as anti-angiogenesis agent in future. Copyright © 2016 Elsevier Ltd. All rights reserved.

  20. Functional characterization of the transcription factor ZEB1 in epithelial to mesenchymal transition and cancer progression

    International Nuclear Information System (INIS)

    Sultan, A.

    2010-01-01

    Epithelial to mesenchymal transition (EMT) is implicated in the progression of primary tumours towards metastasis and is likely caused by a pathological activation of transcription factors regulating EMT in embryonic development. To analyse EMT-causing pathways in tumourigenesis, transcriptional targets of the E-cadherin repressor ZEB1 in invasive humancancer cells were identified. We show that ZEB1 repressed multiple key determinants of epithelial differentiation and cell-cell adhesion, including the cell polarity genes Crumbs3, HUGL2, PKP3 and Pals1-associated tight junction protein. ZEB1 associated with their endogenous promoters in vivo, and strongly repressed promoter activities in reporter assays. ZEB1 downregulation in undifferentiated cancer cells by RNA interference was sufficient to upregulate expression of these cell polarity genes on the RNA and protein level, to re-establish epithelial features and to impair cell motility in vitro. In human colorectal cancer, ZEB1 expression was limited to the tumour-host interface and was accompanied by loss of intercellular adhesion and tumour cell invasion. EMT-inducing transcriptional repressor ZEB1 promotes colorectal cancer cell metastasis and loss of cell polarity. Thereby, ZEB1 suppresses the expression of cell polarity factors, in particular of Lgl2, which was found to be reduced in colorectal and breast cancers. In invasive ductal and lobular breast cancer, upregulation of ZEB1 was stringently coupled to cancer cell dedifferentiation. The invasion potential of MDA-MB-231, a highly invasive breast cancer cell line, is shown to be under the control of ZEB1. Over-expression of ZEB1downregulates and relocalizes E-Cadherin in MCF7 breast cancer cells; moreover, ZEB1 overexpression results in reduced proliferation rate of these cells. Most importantly, we show that ZEB1 mediated downregulation of E-cadherin involves chromatin modifications. Markers of transcriptionally active chromatin Acetylated H3 and Acetylated

  1. Characterization of receptors for recombinant human tumor necrosis factor-alpha from human placental membranes

    International Nuclear Information System (INIS)

    Aiyer, R.A.; Aggarwal, B.B.

    1990-01-01

    High affinity receptors for recombinant human tumor necrosis factor-alpha (rhTNF-alpha) were identified on membranes prepared from full term human placenta. Highly purified rhTNF-alpha iodinated by the iodogen method was found to bind placental membranes in a displaceable manner with an approximate dissociation constant (KD) of 1.9 nM. The membrane bound TNF-alpha receptor could be solubilized by several detergents with optimum extraction being obtained with 1% Triton X-100. The binding of 125I-rhTNF-alpha to the solubilized receptor was found to be time and temperature dependent, yielding maximum binding within 1 h, 24 h and 48 h at 37 degrees C, 24 degrees C and 4 degrees C, respectively. However, the maximum binding obtainable at 4 degrees C was only 40% of that at 37 degrees C. The binding 125I-rhTNF-alpha to solubilized placental membrane extracts was displaceable by unlabeled rhTNF-alpha, but not by a related protein recombinant human tumor necrosis factor-beta (rhTNF-beta; previously called lymphotoxin). This is similar to the behavior of TNF-alpha receptors derived from detergent-solubilized cell extracts, although on intact cells, both rhTNF-alpha and rhTNF-beta bind with equal affinity to TNF receptors. The Scatchard analysis of the binding data of the solubilized receptor revealed high affinity binding sites with a KD of approximately 0.5 nM and a receptor concentration of about 1 pmole/mg protein. Gel filtration of the solubilized receptor-ligand complexes on Sephacryl S-300 revealed two different peaks of radioactivity at approximate molecular masses of 50,000 Da and 400,000 Da. The 400,000 dalton peak corresponded to the receptor-ligand complex. Overall, our results suggest that high affinity receptors for TNF-alpha are present on human placental membranes and provide evidence that these receptors may be different from that of rhTNF-beta

  2. Characterization of Zebrafish von Willebrand Factor Reveals Conservation of Domain Structure, Multimerization, and Intracellular Storage

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    Arunima Ghosh

    2012-01-01

    Full Text Available von Willebrand disease (VWD is the most common inherited human bleeding disorder and is caused by quantitative or qualitative defects in von Willebrand factor (VWF. VWF is a secreted glycoprotein that circulates as large multimers. While reduced VWF is associated with bleeding, elevations in overall level or multimer size are implicated in thrombosis. The zebrafish is a powerful genetic model in which the hemostatic system is well conserved with mammals. The ability of this organism to generate thousands of offspring and its optical transparency make it unique and complementary to mammalian models of hemostasis. Previously, partial clones of zebrafish vwf have been identified, and some functional conservation has been demonstrated. In this paper we clone the complete zebrafish vwf cDNA and show that there is conservation of domain structure. Recombinant zebrafish Vwf forms large multimers and pseudo-Weibel-Palade bodies (WPBs in cell culture. Larval expression is in the pharyngeal arches, yolk sac, and intestinal epithelium. These results provide a foundation for continued study of zebrafish Vwf that may further our understanding of the mechanisms of VWD.

  3. Molecular dynamics characterization of five pathogenic factor X mutants associated with decreased catalytic activity

    KAUST Repository

    Abdel-Azeim, Safwat

    2014-11-11

    Factor X (FX) is one of the major players in the blood coagulation cascade. Upon activation to FXa, it converts prothrombin to thrombin, which in turn converts fibrinogen into fibrin (blood clots). FXa deficiency causes hemostasis defects, such as intracranial bleeding, hemathrosis, and gastrointestinal blood loss. Herein, we have analyzed a pool of pathogenic mutations, located in the FXa catalytic domain and directly associated with defects in enzyme catalytic activity. Using chymotrypsinogen numbering, they correspond to D102N, T135M, V160A, G184S, and G197D. Molecular dynamics simulations were performed for 1.68 μs on the wild-type and mutated forms of FXa. Overall, our analysis shows that four of the five mutants considered, D102N, T135M, V160A, and G184S, have rigidities higher than those of the wild type, in terms of both overall protein motion and, specifically, subpocket S4 flexibility, while S1 is rather insensitive to the mutation. This acquired rigidity can clearly impact the substrate recognition of the mutants.

  4. Characterization of the Factors that Influence Sinapine Concentration in Rapeseed Meal during Fermentation

    Science.gov (United States)

    Niu, Yanxing; Jiang, Mulan; Guo, Mian; Wan, Chuyun; Hu, Shuangxi; Jin, Hu; Huang, Fenghong

    2015-01-01

    We analyzed and compared the difference in sinapine concentration in rapeseed meal between the filamentous fungus, Trametes sp 48424, and the yeast, Saccharomyces cerevisiae, in both liquid and solid-state fermentation. During liquid and solid-state fermentation by Trametes sp 48424, the sinapine concentration decreased significantly. In contrast, the liquid and solid-state fermentation process by Saccharomyces cerevisiae just slightly decreased the sinapine concentration (P ≤ 0.05). After the solid-state fermented samples were dried, the concentration of sinapine in rapeseed meal decreased significantly in Saccharomyces cerevisiae. Based on the measurement of laccase activity, we observed that laccase induced the decrease in the concentration of sinapine during fermentation with Trametes sp 48424. In order to eliminate the influence of microorganisms and the metabolites produced during fermentation, high moisture rapeseed meal and the original rapeseed meal were dried at 90°C and 105°C, respectively. During drying, the concentration of sinapine in high moisture rapeseed meal decreased rapidly and we obtained a high correlation coefficient between the concentration of sinapine and loss of moisture. Our results suggest that drying and enzymes, especially laccase that is produced during the solid-state fermentation process, may be the main factors that affect the concentration of sinapine in rapeseed meal. PMID:25606856

  5. Characterization of the Methylation Status of Pax7 and Myogenic Regulator Factors in Cell Myogenic Differentiation.

    Science.gov (United States)

    Chao, Zhe; Zheng, Xin-Li; Sun, Rui-Ping; Liu, Hai-Long; Huang, Li-Li; Cao, Zong-Xi; Deng, Chang-Yan; Wang, Feng

    2016-07-01

    Epigenetic processes in the development of skeletal muscle have been appreciated for over a decade. DNA methylation is a major epigenetic modification important for regulating gene expression and suppressing spurious transcription. Up to now, the importance of epigenetic marks in the regulation of Pax7 and myogenic regulatory factors (MRFs) expression is far less explored. In the present study, semi-quantitative the real-time polymerase chain reaction (RT-PCR) analyses showed MyoD and Myf5 were expressed in activated and quiescent C2C12 cells. MyoG was expressed in a later stage of myogenesis. Pax7 was weakly expressed in differentiated C2C12 cells. To further understand the regulation of expression of these genes, the DNA methylation status of Pax7, MyoD, and Myf5 was determined by bisulfite sequencing PCR. During the C2C12 myoblasts fusion process, the changes of promoter and exon 1 methylation of Pax7, MyoD, and Myf5 genes were observed. In addition, an inverse relationship of low methylation and high expression was found. These results suggest that DNA methylation may be an important mechanism regulating Pax7 and MRFs transcription in cell myogenic differentiation.

  6. Molecular dynamics characterization of five pathogenic factor X mutants associated with decreased catalytic activity

    KAUST Repository

    Abdel-Azeim, Safwat; Oliva, Romina M.; Chermak, Edrisse; De Cristofaro, Raimondo; Cavallo, Luigi

    2014-01-01

    Factor X (FX) is one of the major players in the blood coagulation cascade. Upon activation to FXa, it converts prothrombin to thrombin, which in turn converts fibrinogen into fibrin (blood clots). FXa deficiency causes hemostasis defects, such as intracranial bleeding, hemathrosis, and gastrointestinal blood loss. Herein, we have analyzed a pool of pathogenic mutations, located in the FXa catalytic domain and directly associated with defects in enzyme catalytic activity. Using chymotrypsinogen numbering, they correspond to D102N, T135M, V160A, G184S, and G197D. Molecular dynamics simulations were performed for 1.68 μs on the wild-type and mutated forms of FXa. Overall, our analysis shows that four of the five mutants considered, D102N, T135M, V160A, and G184S, have rigidities higher than those of the wild type, in terms of both overall protein motion and, specifically, subpocket S4 flexibility, while S1 is rather insensitive to the mutation. This acquired rigidity can clearly impact the substrate recognition of the mutants.

  7. Selection and characterization of a human neutralizing antibody to human fibroblast growth factor-2

    International Nuclear Information System (INIS)

    Tao, Jun; Xiang, Jun-Jian; Li, Dan; Deng, Ning; Wang, Hong; Gong, Yi-Ping

    2010-01-01

    Compelling evidences suggest that fibroblast growth factor-2 (FGF-2) plays important roles in tumor growth, angiogenesis and metastasis. Molecules blocking the FGF-2 signaling have been proposed as anticancer agents. Through screening of a human scFv phage display library, we have isolated several human single-chain Fv fragments (scFvs) that bind to human FGF-2. After expression and purification in bacteria, one scFv, named 1A2, binds to FGF-2 with a high affinity and specificity, and completes with FGF-2 binding to its receptor. This 1A2 scFv was then cloned into the pIgG1 vector and expressed in 293T cells. The purified hIgG1-1A2 antibody showed a high binding affinity of 8 x 10 -9 M to rhFGF-2. In a set of vitro assays, it inhibited various biological activities of FGF-2 such as the proliferation, migration and tube formation of human umbilical vein endothelial cells. More importantly, hIgG1-1A2 antibody also efficiently blocked the growth while inducing apoptosis of glioma cells. For the first time, we generated a human anti-FGF-2 antibody with proven in vitro anti-tumor activity. It may therefore present a new therapeutic candidate for the treatment of cancers that are dependent on FGF-2 signaling for growth and survival.

  8. High Efficient Expression, Purification, and Functional Characterization of Native Human Epidermal Growth Factor in Escherichia coli

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    Yi Ma

    2016-01-01

    Full Text Available Human epidermal growth factor (hEGF is a small, mitotic growth polypeptide that promotes the proliferation of various cells and is widely applied in clinical practices. However, high efficient expression of native hEGF in Escherichia coli has not been successful, since three disulfide bonds in monomer hEGF made it unable to fold into correct 3D structure using in vivo system. To tackle this problem, we fused Mxe GyrA intein (Mxe at the C-terminal of hEGF followed by small ubiquitin-related modifier (SUMO and 10x His-tag to construct a chimeric protein hEGF-Mxe-SUMO-H10. The fusion protein was highly expressed at the concentration of 281 mg/L and up to 59.5% of the total cellular soluble proteins. The fusion protein was purified by affinity chromatography and 29.4 mg/L of native hEGF can be released by thiol induced N-terminal cleavage without any proteases. The mitotic activity in Balb/c 3T3 cells is proliferated by commercial and recombinant hEGF measured with methylthiazolyldiphenyl-tetrazolium bromide (MTT assay which indicated that recombinant hEGF protein stimulates the cell proliferation similar to commercial protein. This study significantly improved the yield and reduced the cost of hEGF in the recombinant E. coli system and could be a better strategy to produce native hEGF for pharmaceutical development.

  9. High Efficient Expression, Purification, and Functional Characterization of Native Human Epidermal Growth Factor in Escherichia coli.

    Science.gov (United States)

    Ma, Yi; Yu, Jieying; Lin, Jinglian; Wu, Shaomin; Li, Shan; Wang, Jufang

    2016-01-01

    Human epidermal growth factor (hEGF) is a small, mitotic growth polypeptide that promotes the proliferation of various cells and is widely applied in clinical practices. However, high efficient expression of native hEGF in Escherichia coli has not been successful, since three disulfide bonds in monomer hEGF made it unable to fold into correct 3D structure using in vivo system. To tackle this problem, we fused Mxe GyrA intein (Mxe) at the C-terminal of hEGF followed by small ubiquitin-related modifier (SUMO) and 10x His-tag to construct a chimeric protein hEGF-Mxe-SUMO-H 10 . The fusion protein was highly expressed at the concentration of 281 mg/L and up to 59.5% of the total cellular soluble proteins. The fusion protein was purified by affinity chromatography and 29.4 mg/L of native hEGF can be released by thiol induced N-terminal cleavage without any proteases. The mitotic activity in Balb/c 3T3 cells is proliferated by commercial and recombinant hEGF measured with methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay which indicated that recombinant hEGF protein stimulates the cell proliferation similar to commercial protein. This study significantly improved the yield and reduced the cost of hEGF in the recombinant E. coli system and could be a better strategy to produce native hEGF for pharmaceutical development.

  10. Structural characterization of the virulence factor nuclease A from Streptococcus agalactiae.

    Science.gov (United States)

    Moon, Andrea F; Gaudu, Philippe; Pedersen, Lars C

    2014-11-01

    The group B pathogen Streptococcus agalactiae commonly populates the human gut and urogenital tract, and is a major cause of infection-based mortality in neonatal infants and in elderly or immunocompromised adults. Nuclease A (GBS_NucA), a secreted DNA/RNA nuclease, serves as a virulence factor for S. agalactiae, facilitating bacterial evasion of the human innate immune response. GBS_NucA efficiently degrades the DNA matrix component of neutrophil extracellular traps (NETs), which attempt to kill and clear invading bacteria during the early stages of infection. In order to better understand the mechanisms of DNA substrate binding and catalysis of GBS_NucA, the high-resolution structure of a catalytically inactive mutant (H148G) was solved by X-ray crystallography. Several mutants on the surface of GBS_NucA which might influence DNA substrate binding and catalysis were generated and evaluated using an imidazole chemical rescue technique. While several of these mutants severely inhibited nuclease activity, two mutants (K146R and Q183A) exhibited significantly increased activity. These structural and biochemical studies have greatly increased our understanding of the mechanism of action of GBS_NucA in bacterial virulence and may serve as a foundation for the structure-based drug design of antibacterial compounds targeted to S. agalactiae.

  11. Tumor necrosis factor alpha of teleosts: in silico characterization and homology modeling

    Directory of Open Access Journals (Sweden)

    Tran Ngoc Tuan

    2016-10-01

    Full Text Available Tumor necrosis factor alpha (TNF- is known to be crucial in many biological activities of organisms. In this study, physicochemical properties and modeling of TNF- protein of fish was analyzed using in silico approach. TNF- proteins selected from fish species, including grass carp (Ctenopharyngodon idella, zebra fish (Danio rerio, Nile tilapia (Oreochromis niloticus, goldfish (Carassius auratus, and rainbow trout (Oncorhynchus mykiss were used in this study. Physicochemical characteristics with molecular weight, theoretical isoelectric point, extinction coefficient, aliphatic index, instability index, total number of negatively charged residues and positively charged residues, and grand average of hydropathicity were computed. All proteins were classified as transmembrane proteins. The “transmembrane region” and “TNF” domain were identified from protein sequences. The function prediction of proteins was also performed. Alpha helices and random coils were dominating in the secondary structure of the proteins. Three-dimensional structures were predicted and verified as good structures for the investigation of TNF- of fish by online server validation.

  12. The glycosylation and characterization of the candidate Gc macrophage activating factor.

    Science.gov (United States)

    Ravnsborg, Tina; Olsen, Dorthe T; Thysen, Anna Hammerich; Christiansen, Maja; Houen, Gunnar; Højrup, Peter

    2010-04-01

    The vitamin D binding protein, Gc globulin, has in recent years received some attention for its role as precursor for the extremely potent macrophage activating factor (GcMAF). An O-linked trisaccharide has been allocated to the threonine residue at position 420 in two of the three most common isoforms of Gc globulin (Gc1s and Gc1f). A substitution for a lysine residue at position 420 in Gc2 prevents this isoform from being glycosylated at that position. It has been suggested that Gc globulin subjected sequentially to sialidase and galactosidase treatment generates GcMAF in the form of Gc globulin with only a single GalNAc attached to T420. In this study we confirm the location of a linear trisaccharide on T420. Furthermore, we provide the first structural evidence of the generation of the proposed GcMAF by use of glycosidase treatment and mass spectrometry. Additionally the generated GcMAF candidate was tested for its effect on cytokine release from macrophages in human whole blood. Copyright 2010 Elsevier B.V. All rights reserved.

  13. Characterization of germ cell-specific expression of the orphan nuclear receptor, germ cell nuclear factor.

    Science.gov (United States)

    Katz, D; Niederberger, C; Slaughter, G R; Cooney, A J

    1997-10-01

    Nuclear receptors, such as those for androgens, estrogens, and progesterones, control many reproductive processes. Proteins with structures similar to these receptors, but for which ligands have not yet been identified, have been termed orphan nuclear receptors. One of these orphans, germ cell nuclear factor (GCNF), has been shown to be germ cell specific in the adult and, therefore, may also participate in the regulation of reproductive functions. In this paper, we examine more closely the expression patterns of GCNF in germ cells to begin to define spatio-temporal domains of its activity. In situ hybridization showed that GCNF messenger RNA (mRNA) is lacking in the testis of hypogonadal mutant mice, which lack developed spermatids, but is present in the wild-type testis. Thus, GCNF is, indeed, germ cell specific in the adult male. Quantitation of the specific in situ hybridization signal in wild-type testis reveals that GCNF mRNA is most abundant in stage VII round spermatids. Similarly, Northern analysis and specific in situ hybridization show that GCNF expression first occurs in testis of 20-day-old mice, when round spermatids first emerge. Therefore, in the male, GCNF expression occurs postmeiotically and may participate in the morphological changes of the maturing spermatids. In contrast, female expression of GCNF is shown in growing oocytes that have not completed the first meiotic division. Thus, GCNF in the female is expressed before the completion of meiosis. Finally, the nature of the two different mRNAs that hybridize to the GCNF complementary DNA was studied. Although both messages contain the DNA binding domain, only the larger message is recognized by a probe from the extreme 3' untranslated region. In situ hybridization with these differential probes demonstrates that both messages are present in growing oocytes. In addition, the coding region and portions of the 3' untranslated region of the GCNF complementary DNA are conserved in the rat.

  14. Functional genomic characterization of virulence factors from necrotizing fasciitis-causing strains of Aeromonas hydrophila.

    Science.gov (United States)

    Grim, Christopher J; Kozlova, Elena V; Ponnusamy, Duraisamy; Fitts, Eric C; Sha, Jian; Kirtley, Michelle L; van Lier, Christina J; Tiner, Bethany L; Erova, Tatiana E; Joseph, Sandeep J; Read, Timothy D; Shak, Joshua R; Joseph, Sam W; Singletary, Ed; Felland, Tracy; Baze, Wallace B; Horneman, Amy J; Chopra, Ashok K

    2014-07-01

    The genomes of 10 Aeromonas isolates identified and designated Aeromonas hydrophila WI, Riv3, and NF1 to NF4; A. dhakensis SSU; A. jandaei Riv2; and A. caviae NM22 and NM33 were sequenced and annotated. Isolates NF1 to NF4 were from a patient with necrotizing fasciitis (NF). Two environmental isolates (Riv2 and -3) were from the river water from which the NF patient acquired the infection. While isolates NF2 to NF4 were clonal, NF1 was genetically distinct. Outside the conserved core genomes of these 10 isolates, several unique genomic features were identified. The most virulent strains possessed one of the following four virulence factors or a combination of them: cytotoxic enterotoxin, exotoxin A, and type 3 and 6 secretion system effectors AexU and Hcp. In a septicemic-mouse model, SSU, NF1, and Riv2 were the most virulent, while NF2 was moderately virulent. These data correlated with high motility and biofilm formation by the former three isolates. Conversely, in a mouse model of intramuscular infection, NF2 was much more virulent than NF1. Isolates NF2, SSU, and Riv2 disseminated in high numbers from the muscular tissue to the visceral organs of mice, while NF1 reached the liver and spleen in relatively lower numbers on the basis of colony counting and tracking of bioluminescent strains in real time by in vivo imaging. Histopathologically, degeneration of myofibers with significant infiltration of polymorphonuclear cells due to the highly virulent strains was noted. Functional genomic analysis provided data that allowed us to correlate the highly infectious nature of Aeromonas pathotypes belonging to several different species with virulence signatures and their potential ability to cause NF. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  15. Functional characterization of Bombyx mori nucleopolyhedrovirus mutant lacking late expression factor 9.

    Science.gov (United States)

    Zhang, Y; Shi, Y; Yu, H; Li, J; Quan, Y; Shu, T; Nie, Z; Zhang, Y; Yu, W

    Baculoviridae is a family of invertebrate viruses with large double-stranded DNA genomes. Proteins encoded by some late expression factor (lef ) genes are involved in the regulation of viral gene expression. Lef-9 is one of four transcription-specific Lefs, which are components of the virus-encoded RNA polymerase, and can initiate and transcribe late and very late genes. As a multifunctional protein encoded by the Bombyx mori nucleopolyhedrovirus (BmNPV), Lef-9 may be involved in the regulation of viral propagation. However, the underlying mechanism remains unclear. To determine the role of lef-9 in baculovirus infection, lef-9-knockout virus (lef-9-KO-Bacmid virus) was constructed using the Red recombination system, and the Bac-to-Bac system was used to prepare lef-9-repaired virus (lef-9-Re-Bacmid virus). The lef-9-KO virus did not produce infectious viruses or show infection activity, while the lef-9-repaired virus recovered both. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis of the transcription levels in wild-type-Bacmid, lef-9-KO-Bacmid, and lef-9-Re-Bacmid viruses showed that the lef-9-KO bacmid had little effect on viral genome replication. However, the transcription levels of the early and late viral genes, lef-3, ie-1, vp39, and p10, were significantly lower in BmN cells transfected with lef-9-KO-Bacmids than in the controls. Electron microscopy showed no visible enveloped virions in cells transfected with lef-9-KO-Bacmids, while many mature virions in cells transfected with lef-9-Re-Bacmid and wt-Bacmid were present. Thus, lef-9 was not essential for viral genome replication, but significantly affected viral gene transcription and expression in all periods of cell life cycle.

  16. Synthesis, characterization, vibrational spectroscopy, and factor group analysis of partially metal-doped phosphate materials

    Science.gov (United States)

    Sronsri, Chuchai; Boonchom, Banjong

    2018-04-01

    A simple precipitating method was used to synthesize effectively a partially metal-doped phosphate hydrate (Mn0.9Mg0.1HPO4·3H2O), whereas the thermal decomposition process of the above hydrate precursor was used to obtain Mn1.8Mg0.2P2O7 and LiMn0.9Mg0.1PO4 compounds under different conditions. To separate the overlapping thermal decomposition peak, a deconvolution technique was used, and the separated peak was applied to calculate the water content. The factor group splitting analysis was used to exemplify their vibrational spectra obtained from normal vibrations of HPO42-, H2O, P2O74- and PO43- functional groups. Further, the deconvoluted bending mode of water was clearly observed. Mn0.9Mg0.1HPO4·3H2O was observed in the orthorhombic crystal system with the space group of Pbca (D2h15). The formula units per unit cell were found to be eight (Z = 8), and the site symmetric type of HPO42- was observed as Cs. For the HPO42- unit, the correlation filed splitting analysis of type C3v - Cs - D2h15 was calculated and had 96 internal modes, whereas H2O in the above hydrate was symbolized as C2v - Cs - D2h15 and had 24 modes. The symbol C2v - Cs - C2h3 was used for the correlation filed splitting analysis of P2O74- in Mn1.8Mg0.2P2O7 (monoclinic, C2/m (C2h3), Z = 2, and 42 modes). Finally, the symbol Td - Cs - D2h16 was used for the correlation filed splitting analysis of PO43- in LiMn0.9Mg0.1PO4 (orthorhombic, Pnma (D2h16), Z = 4, and 36 modes).

  17. Structural and functional characterization of the product of disease-related factor H gene conversion.

    Science.gov (United States)

    Herbert, Andrew P; Kavanagh, David; Johansson, Conny; Morgan, Hugh P; Blaum, Bärbel S; Hannan, Jonathan P; Barlow, Paul N; Uhrín, Dušan

    2012-03-06

    Numerous complement factor H (FH) mutations predispose patients to atypical hemolytic uremic syndrome (aHUS) and other disorders arising from inadequately regulated complement activation. No unifying structural or mechanistic consequences have been ascribed to these mutants beyond impaired self-cell protection. The S1191L and V1197A mutations toward the C-terminus of FH, which occur in patients singly or together, arose from gene conversion between CFH encoding FH and CFHR1 encoding FH-related 1. We show that neither single nor double mutations structurally perturbed recombinant proteins consisting of the FH C-terminal modules, 19 and 20 (FH19-20), although all three FH19-20 mutants were poor, compared to wild-type FH19-20, at promoting hemolysis of C3b-coated erythrocytes through competition with full-length FH. Indeed, our new crystal structure of the S1191L mutant of FH19-20 complexed with an activation-specific complement fragment, C3d, was nearly identical to that of the wild-type FH19-20:C3d complex, consistent with mutants binding to C3b with wild-type-like affinity. The S1191L mutation enhanced thermal stability of module 20, whereas the V1197A mutation dramatically decreased it. Thus, although mutant proteins were folded at 37 °C, they differ in conformational rigidity. Neither single substitutions nor double substitutions increased measurably the extent of FH19-20 self-association, nor did these mutations significantly affect the affinity of FH19-20 for three glycosaminoglycans, despite critical roles of module 20 in recognizing polyanionic self-surface markers. Unexpectedly, FH19-20 mutants containing Leu1191 self-associated on a heparin-coated surface to a higher degree than on surfaces coated with dermatan or chondroitin sulfates. Thus, potentially disease-related functional distinctions between mutants, and between FH and FH-related 1, may manifest in the presence of specific glycosaminoglycans.

  18. Prevalence, risk factors of infection and molecular characterization of trichomonads in puppies from French breeding kennels.

    Science.gov (United States)

    Grellet, Aurélien; Brunopolack; Feugier, Alexandre; Boucraut-Baralon, Corine; Grandjean, Dominique; Vandewynckel, Laurine; Cian, Amandine; Meloni, Dionigia; Viscogliosi, Eric

    2013-11-08

    The trichomonad species Tritrichomonas fetus and Pentatrichomonas hominis were recently identified in the feces of dogs with diarrhea. However the prevalence and pathogenicity of these parasites in the canine population still remained poorly resolved. Therefore the aim of the present study was (1) to determine the prevalence of trichomonads infecting puppies living in French breeding kennels, (2) to confirm the predominance of P. hominis in dogs, (3) to investigate the genetic diversity of P. hominis isolates identified in the French canine population and (4) to evaluate the risk factors for infection by P. hominis and the influence of the parasite on feces consistency. A total of 215 both diarrheic and non-diarrheic puppies from 25 French breeding kennels were included in this epidemiological survey. Fecal samples from each puppy were examined for 6 gastrointestinal pathogens: parvovirus type 2 (CPV2), coronavirus, Toxocara canis, Cystoisospora ohioensis-complex, Cystoisospora canis, and Giardia intestinalis. A part of each collected stool was also tested for the presence of motile trichomonads by microscopy after culturing. The prevalence of trichomonad infection was 15.8% (34/215) among puppies and 20% (5/25) among breeding kennels. DNA from 26 of the 34 positive samples was successfully amplified using a trichomonad-specific primer pair. Analysis of the sequences of PCR products indicated that P. hominis was the only trichomonad infecting the canine population. All the puppies infected with P. hominis belonged to large breed dogs. Moreover, puppies from large breeding kennels, excreting a high level of G. intestinalis and/or excreting a high level of C. canis oocysts showed a higher probability of being positive for P. hominis infection. Univariate analysis also revealed an increased risk for P. hominis infection in puppies with abnormal feces. However, in a multivariate analysis, CPV2 was the only gastrointestinal pathogen associated with abnormal feces. Since

  19. Using NASA Remotely Sensed Data to Help Characterize Environmental Risk Factors for National Public Health Applications

    Science.gov (United States)

    Al-Hamdan, M. Z.; Crosson, W. L.; Economou, S.; Estes, M., Jr.; Estes, S. M.; Hemmings, S. N.; Kent, S.; Loop, M.; Puckett, M.; Quattrochi, D. A.; Wade, G.; McClure, L.

    2012-12-01

    The overall goal of this study is to address issues of environmental health and enhance public health decision making by using NASA remotely sensed data and products. This study is a collaboration between NASA Marshall Space Flight Center, Universities Space Research Association (USRA), the University of Alabama at Birmingham (UAB) School of Public Health and the Centers for Disease Control and Prevention (CDC) Office of Surveillance, Epidemiology and Laboratory Services. The objectives of this study are to develop high-quality spatial data sets of environmental variables, link these with public health data from a national cohort study, and deliver the environmental data sets and associated public health analyses to local, state and federal end-user groups. Three daily environmental data sets were developed for the conterminous U.S. on different spatial resolutions for the period 2003-2008: (1) spatial surfaces of estimated fine particulate matter (PM2.5) on a 10-km grid using US Environmental Protection Agency (EPA) ground observations and NASA's MODerate-resolution Imaging Spectroradiometer (MODIS) data; (2) a 1-km grid of MODIS Land Surface Temperature (LST); and (3) a 12-km grid of daily incoming solar radiation and maximum and minimum air temperature using the North American Land Data Assimilation System (NLDAS) data. These environmental datasets were linked with public health data from the UAB REasons for Geographic and Racial Differences in Stroke (REGARDS) national cohort study to determine whether exposures to these environmental risk factors are related to cognitive decline, stroke and other health outcomes. These environmental national datasets will also be made available to public health professionals, researchers and the general public via the CDC Wide-ranging Online Data for Epidemiologic Research (WONDER) system, where they can be aggregated to the county-level, state-level, or regional-level as per users' need and downloaded in tabular, graphical

  20. Establishment and characterization of a new and stable collagen-binding assay for the assessment of von Willebrand factor activity

    Science.gov (United States)

    Ni, Y; Nesrallah, J; Agnew, M; Geske, F J; Favaloro, E J

    2013-01-01

    Introduction Laboratory diagnosis of von Willebrand disease (VWD) requires determination of both von Willebrand factor (VWF) protein levels and activity. Current VWF activity tests include the ristocetin cofactor assay and the collagen-binding assay (VWF:CB). The goal of this investigation is to characterize a new collagen-binding assay and to determine its effectiveness in identifying VWD. Methods Analytical studies were carried out to characterize the performance of a new VWF:CB ELISA. Additionally, samples from a normal population were tested as were well-characterized type 1 and type 2 VWD samples. Results Repeatability and within-laboratory precision studies resulted in coefficients of variation (CVs) of ≤11%. A linear range of 1–354% (0.01–3.54 IU/mL) was determined, along with a limit of detection and a lower limit of quantitation of 1.6% and 4.0% (0.016 and 0.04 IU/mL), respectively. Samples tested from apparently healthy individuals resulted in a normal range of 54–217% (0.54–2.17 IU/mL). Known VWD type 1 and type 2 samples were also analyzed by the ELISA, with 99% of samples having VWF:CB below the normal reference range and an estimated 96% sensitivity and 87% specificity using a VWF collagen-binding/antigen cutoff ratio of 0.50. Conclusion This new VWF:CB ELISA provides an accurate measure of collagen-binding activity that aids in the diagnosis and differentiation of type 1 from type 2 VWD. PMID:23107512

  1. Characterization of a canine tetranucleotide microsatellite marker located in the first intron of the tumor necrosis factor alpha gene.

    Science.gov (United States)

    Watanabe, Masashi; Tanaka, Kazuaki; Takizawa, Tatsuya; Segawa, Kazuhito; Neo, Sakurako; Tsuchiya, Ryo; Murata, Michiko; Murakami, Masaru; Hisasue, Masaharu

    2014-01-01

    A polymorphic tetranucleotide (GAAT)n microsatellite in the first intron of the canine tumor necrosis factor alpha (TNFA) gene was characterized in this study; 139 dogs were analyzed: 22 Beagles, 26 Chihuahuas, 20 Miniature Dachshunds, 24 Miniature Poodles, 22 Pembroke Welsh Corgis and 25 Shiba Inus. We detected the presence of the 4 alleles (GAAT)5, (GAAT)6, (GAAT)7 and (GAAT)8, including 9 of the 10 expected genotypes. The expected heterozygosity (He) and the polymorphic information content (PIC) value of this microsatellite locus varied from 0.389 to 0.749 and from 0.333 to 0.682, respectively, among the 6 breeds. The allelic frequency differed greatly among breeds, but this microsatellite marker was highly polymorphic and could be a useful marker for the canine TNFA gene.

  2. Molecular characterization of the mouse superior lateral parabrachial nucleus through expression of the transcription factor Runx1.

    Directory of Open Access Journals (Sweden)

    Chrissandra J Zagami

    2010-11-01

    Full Text Available The ability to precisely identify separate neuronal populations is essential to the understanding of the development and function of different brain structures. This necessity is particularly evident in regions such as the brainstem, where the anatomy is quite complex and little is known about the identity, origin, and function of a number of distinct nuclei due to the lack of specific cellular markers. In this regard, the gene encoding the transcription factor Runx1 has emerged as a specific marker of restricted neuronal populations in the murine central and peripheral nervous systems. The aim of this study was to precisely characterize the expression of Runx1 in the developing and postnatal mouse brainstem.Anatomical and immunohistochemical studies were used to characterize mouse Runx1 expression in the brainstem. It is shown here that Runx1 is expressed in a restricted population of neurons located in the dorsolateral rostral hindbrain. These neurons define a structure that is ventromedial to the dorsal nucleus of the lateral lemniscus, dorsocaudal to the medial paralemniscal nucleus and rostral to the cerebellum. Runx1 expression in these cells is first observed at approximately gestational day 12.5, persists into the adult brain, and is lost in knockout mice lacking the transcription factor Atoh1, an important regulator of the development of neuronal lineages of the rhombic lip. Runx1-expressing neurons in the rostral hindbrain produce cholecystokinin and also co-express members of the Groucho/Transducin-like Enhancer of split protein family.Based on the anatomical and molecular characteristics of the Runx1-expressing cells in the rostral hindbrain, we propose that Runx1 expression in this region of the mouse brain defines the superior lateral parabrachial nucleus.

  3. Ammonium Oxidation Under Iron Reducing Conditions: Environmental Factors Characterization and Process Optimization

    Science.gov (United States)

    Huang, Shan; Ruiz, Melany; Jaffe, Peter

    2015-04-01

    Ammonium (NH4+) oxidation coupled to iron (Fe) reduction in the absence of oxygen and nitrate/nitrite (NO3-/NO2-) has been reported by several investigators and is referred to as Feammox. Feammox is a biological reaction, where Fe(III) is the electron acceptor, which is reduced to Fe(II), and NH4+ is the electron donor, which is oxidized to NO2-. An Acidimicrobiaceae bacterium named A6, a previously unreported species in the Acidimicrobiaceae family, has been identified as being responsible for the Feammox process(1, 2) Feammox process was noted in riparian wetland soils in New Jersey(1,3), in tropical rainforest soils in Puerto Rico (4) and in paddy soils in China (5). In addition to these published locations, Feammox process was also found in samples collected from a series of local wetland-, upland-, as well as storm-water detention pond-sediments in New Jersey, river sediments from South Carolina, and forested soils near an acid mine drainage (Dabaoshan, Guangdong province) in China. Using primers acm342f - 439r (2), Acidimicrobiaceae bacterium A6 was detected in samples where Feammox was observed, after strictly anaerobic incubations. According to a canonical correspondence analysis with environmental characteristics and soil microbial communities, the species-environment relationship indicated that pH and Fe oxides content were the primary factors controlling Feammox process. Anaerobic incubations of Feammox enrichment cultures adjusted to different pH, revealed that the optimal pH for Feammox is 4 ~ 5, and the reaction does not proceed when pH > 7. No correlation was found between the distributions of Feammox bacteria and other NH4+ oxidation bacteria. Pure Acidimicrobiaceae bacterium A6 strain was isolated in an autotrophic medium, from an active Feammox membrane reactor (A6 was enriched to 65.8% of the total bacteria). A 13C labeled CO2 amendment was conducted, and the 13C in cells of A6 increased from 1.80% to 10.3% after 14 days incubation. In a separate

  4. Characterizing Factors Associated with Built-Up Land Expansion in Urban and Non-Urban Areas from a Morphological Perspective

    Directory of Open Access Journals (Sweden)

    Zhonghao Zhang

    2017-08-01

    Full Text Available In this paper, built-up land expansion patterns and the associated factors were characterized in urban and non-urban areas across the Wen-Tai region of eastern China. Fractal dimension can be used as a reliable indicator of the complexity of built-up land form, and the increasing trend of fractal dimension indicated a more complex, dispersed pattern of built-up land in urban areas. Spatial regression models were quantitatively implemented to identify the indicators influencing the variation of fractal dimensions. Our findings suggested that the fractal dimension of built-up land forms was positively correlated to the patch density and elevation when built-up land expansion was more concentrated. Both landscape shape index and Gross Domestic Product (GDP were positively correlated with fractal dimension in urban areas, and total edge, edge density, and connective index had impacts on fractal dimension in non-urban areas. Slope and agricultural population also showed an influence on fractal dimension. This study provided a new way for urban studies in interpreting the complex interactions between fractal dimension and related factors. The combined approach of fractal dimension and spatial analysis can provide the government planners with valuable information that can be efficiently used to realize the influences of land use policies in urban and non-urban areas.

  5. Identification and Characterization of the Diverse Stress-Responsive R2R3-RMYB Transcription Factor from Hibiscus sabdariffa L.

    Science.gov (United States)

    Mohamed, Bahaeldeen Babikar; Aftab, Beenish; Sarwar, Muhammad Bilal; Ahmad, Zarnab; Hassan, Sameera; Husnain, Tayyab

    2017-01-01

    Various regulatory proteins play a fundamental role to manage the healthy plant growth under stress conditions. Differential display reverse transcriptase PCR and random amplification of cDNA ends (RACE) was used to explore the osmotic stress-responsive transcripts. We identified and characterized the salt stress-responsive R2R3 type RMYB transcription factor from Hibiscus sabdariffa which has an open reading frame of 690 bp, encoding 229 long chain amino acids. In silico analysis confirmed the conserved R2 and R3 domain as well as an NLS-1 localization site. The deduced amino acids of RMYB shared 83, 81, 80, 79, 72, 71, and 66% homology with Arabidopsis thaliana, Glycine max, Oryza sativa, Zea maize, Malus domestica, Populus tremula × Populus alba, and Medicago sativa specific MYB family, respectively. We observed the gene upregulation in stem, leaf, and root tissue in response to abiotic stress. Furthermore, RMYB gene was cloned into plant expression vector under CaMV35S promoter and transformed to Gossypium hirsutum: a local cotton cultivar. Overexpression of RMYB was observed in transgenic plants under abiotic stresses which further suggests its regulatory role in response to stressful conditions. The RMYB transcription factor-overexpressing in transgenic cotton plants may be used as potential agent for the development of stress tolerant crop cultivars. PMID:29181384

  6. Characterization of genetically engineered mouse hepatoma cells with inducible liver functions by overexpression of liver-enriched transcription factors.

    Science.gov (United States)

    Yamamoto, Hideaki; Tonello, Jane Marie; Sambuichi, Takanori; Kawabe, Yoshinori; Ito, Akira; Kamihira, Masamichi

    2018-01-01

    New cell sources for the research and therapy of organ failure could significantly alleviate the shortage of donor livers that are available to patients who suffer from liver disease. Liver carcinoma derived cells, or hepatoma cells, are the ideal cells for developing bioartificial liver systems. Such cancerous liver cells are easy to prepare in large quantities and can be maintained over long periods under standard culture conditions, unlike primary hepatocytes. However, hepatoma cells possess only a fraction of the functions of primary hepatocytes. In a previous study, by transducing cells with liver-enriched transcription factors that could be inducibly overexpressed-hepatocyte nuclear factor (HNF)1α, HNF1β, HNF3β [FOXA2], HNF4α, HNF6, CCAAT/enhancer binding protein (C/EBP)α, C/EBPβ and C/EBPγ-we created mouse hepatoma cells with high liver-specific gene expression called the Hepa/8F5 cell line. In the present study, we performed functional and genetic analyses to characterize the Hepa/8F5 cell line. Further, in three-dimensional cultures, the function of these cells improved significantly compared to parental cells. Ultimately, these cells might become a new resource that can be used in basic and applied hepatic research. Copyright © 2017 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  7. Molecular Characterization of the Schistosoma mansoni Zinc Finger Protein SmZF1 as a Transcription Factor

    Science.gov (United States)

    D'Astolfo, Diego S.; Cardoso, Fernanda C.; Rajão, Matheus A.; Mourão, Marina M.; Gava, Elisandra; Oliveira, Sérgio C.; Macedo, Andréa M.; Machado, Carlos R.; Pena, Sérgio D. J.; Kitten, Gregory T.; Franco, Glória R.

    2009-01-01

    Background During its development, the parasite Schistosoma mansoni is exposed to different environments and undergoes many morphological and physiological transformations as a result of profound changes in gene expression. Characterization of proteins involved in the regulation of these processes is of importance for the understanding of schistosome biology. Proteins containing zinc finger motifs usually participate in regulatory processes and are considered the major class of transcription factors in eukaryotes. It has already been shown, by EMSA (Eletrophoretic Mobility Shift Assay), that SmZF1, a S. mansoni zinc finger (ZF) protein, specifically binds both DNA and RNA oligonucleotides. This suggests that this protein might act as a transcription factor in the parasite. Methodology/Principal Findings In this study we extended the characterization of SmZF1 by determining its subcellular localization and by verifying its ability to regulate gene transcription. We performed immunohistochemistry assays using adult male and female worms, cercariae and schistosomula to analyze the distribution pattern of SmZF1 and verified that the protein is mainly detected in the cells nuclei of all tested life cycle stages except for adult female worms. Also, SmZF1 was heterologously expressed in mammalian COS-7 cells to produce the recombinant protein YFP-SmZF1, which was mainly detected in the nucleus of the cells by confocal microscopy and Western blot assays. To evaluate the ability of this protein to regulate gene transcription, cells expressing YFP-SmZF1 were tested in a luciferase reporter system. In this system, the luciferase gene is downstream of a minimal promoter, upstream of which a DNA region containing four copies of the SmZF1 putative best binding site (D1-3DNA) was inserted. SmZF1 increased the reporter gene transcription by two fold (p≤0.003) only when its specific binding site was present. Conclusion Taken together, these results strongly support the hypothesis

  8. Development and Characterization of a Camelid Single Domain Antibody-Urease Conjugate That Targets Vascular Endothelial Growth Factor Receptor 2.

    Science.gov (United States)

    Tian, Baomin; Wong, Wah Yau; Uger, Marni D; Wisniewski, Pawel; Chao, Heman

    2017-01-01

    Angiogenesis is the process of new blood vessel formation and is essential for a tumor to grow beyond a certain size. Tumors secrete the pro-angiogenic factor vascular endothelial growth factor, which acts upon local endothelial cells by binding to vascular endothelial growth factor receptors (VEGFRs). In this study, we describe the development and characterization of V21-DOS47, an immunoconjugate that targets VEGFR2. V21-DOS47 is composed of a camelid single domain anti-VEGFR2 antibody (V21) and the enzyme urease. The conjugate specifically binds to VEGFR2 and urease converts endogenous urea into ammonia, which is toxic to tumor cells. Previously, we developed a similar antibody-urease conjugate, L-DOS47, which is currently in clinical trials for non-small cell lung cancer. Although V21-DOS47 was designed from parameters learned from the generation of L-DOS47, additional optimization was required to produce V21-DOS47. In this study, we describe the expression and purification of two versions of the V21 antibody: V21H1 and V21H4. Each was conjugated to urease using a different chemical cross-linker. The conjugates were characterized by a panel of analytical techniques, including SDS-PAGE, size exclusion chromatography, Western blotting, and LC-MS E peptide mapping. Binding characteristics were determined by ELISA and flow cytometry assays. To improve the stability of the conjugates at physiologic pH, the pIs of the V21 antibodies were adjusted by adding several amino acid residues to the C-terminus. For V21H4, a terminal cysteine was also added for use in the conjugation chemistry. The modified V21 antibodies were expressed in the E. coli BL21 (DE3) pT7 system. V21H1 was conjugated to urease using the heterobifunctional cross-linker succinimidyl-[( N -maleimidopropionamido)-diethyleneglycol] ester (SM(PEG) 2 ), which targets lysine resides in the antibody. V21H4 was conjugated to urease using the homobifunctional cross-linker, 1,8-bis(maleimido)diethylene glycol

  9. Development and Characterization of a Camelid Single Domain Antibody–Urease Conjugate That Targets Vascular Endothelial Growth Factor Receptor 2

    Directory of Open Access Journals (Sweden)

    Baomin Tian

    2017-08-01

    Full Text Available Angiogenesis is the process of new blood vessel formation and is essential for a tumor to grow beyond a certain size. Tumors secrete the pro-angiogenic factor vascular endothelial growth factor, which acts upon local endothelial cells by binding to vascular endothelial growth factor receptors (VEGFRs. In this study, we describe the development and characterization of V21-DOS47, an immunoconjugate that targets VEGFR2. V21-DOS47 is composed of a camelid single domain anti-VEGFR2 antibody (V21 and the enzyme urease. The conjugate specifically binds to VEGFR2 and urease converts endogenous urea into ammonia, which is toxic to tumor cells. Previously, we developed a similar antibody–urease conjugate, L-DOS47, which is currently in clinical trials for non-small cell lung cancer. Although V21-DOS47 was designed from parameters learned from the generation of L-DOS47, additional optimization was required to produce V21-DOS47. In this study, we describe the expression and purification of two versions of the V21 antibody: V21H1 and V21H4. Each was conjugated to urease using a different chemical cross-linker. The conjugates were characterized by a panel of analytical techniques, including SDS-PAGE, size exclusion chromatography, Western blotting, and LC-MSE peptide mapping. Binding characteristics were determined by ELISA and flow cytometry assays. To improve the stability of the conjugates at physiologic pH, the pIs of the V21 antibodies were adjusted by adding several amino acid residues to the C-terminus. For V21H4, a terminal cysteine was also added for use in the conjugation chemistry. The modified V21 antibodies were expressed in the E. coli BL21 (DE3 pT7 system. V21H1 was conjugated to urease using the heterobifunctional cross-linker succinimidyl-[(N-maleimidopropionamido-diethyleneglycol] ester (SM(PEG2, which targets lysine resides in the antibody. V21H4 was conjugated to urease using the homobifunctional cross-linker, 1,8-bis

  10. Characterization of Foodborne Strains of Staphylococcus aureus by Shotgun Proteomics: Functional Networks, Virulence Factors and Species-Specific Peptide Biomarkers

    Science.gov (United States)

    Carrera, Mónica; Böhme, Karola; Gallardo, José M.; Barros-Velázquez, Jorge; Cañas, Benito; Calo-Mata, Pilar

    2017-01-01

    In the present work, we applied a shotgun proteomics approach for the fast and easy characterization of 20 different foodborne strains of Staphylococcus aureus (S. aureus), one of the most recognized foodborne pathogenic bacteria. A total of 644 non-redundant proteins were identified and analyzed via an easy and rapid protein sample preparation procedure. The results allowed the differentiation of several proteome datasets from the different strains (common, accessory, and unique datasets), which were used to determine relevant functional pathways and differentiate the strains into different Euclidean hierarchical clusters. Moreover, a predicted protein-protein interaction network of the foodborne S. aureus strains was created. The whole confidence network contains 77 nodes and 769 interactions. Most of the identified proteins were surface-associated proteins that were related to pathways and networks of energy, lipid metabolism and virulence. Twenty-seven virulence factors were identified, and most of them corresponded to autolysins, N-acetylmuramoyl-L-alanine amidases, phenol-soluble modulins, extracellular fibrinogen-binding proteins and virulence factor EsxA. Potential species-specific peptide biomarkers were screened. Twenty-one species-specific peptide biomarkers, belonging to eight different proteins (nickel-ABC transporter, N-acetylmuramoyl-L-alanine amidase, autolysin, clumping factor A, gram-positive signal peptide YSIRK, cysteine protease/staphopain, transcriptional regulator MarR, and transcriptional regulator Sar-A), were proposed to identify S. aureus. These results constitute the first major dataset of peptides and proteins of foodborne S. aureus strains. This repository may be useful for further studies, for the development of new therapeutic treatments for S. aureus food intoxications and for microbial source-tracking in foodstuffs. PMID:29312172

  11. SNPs altering ammonium transport activity of human Rhesus factors characterized by a yeast-based functional assay.

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    Aude Deschuyteneer

    Full Text Available Proteins of the conserved Mep-Amt-Rh family, including mammalian Rhesus factors, mediate transmembrane ammonium transport. Ammonium is an important nitrogen source for the biosynthesis of amino acids but is also a metabolic waste product. Its disposal in urine plays a critical role in the regulation of the acid/base homeostasis, especially with an acid diet, a trait of Western countries. Ammonium accumulation above a certain concentration is however pathologic, the cytotoxicity causing fatal cerebral paralysis in acute cases. Alteration in ammonium transport via human Rh proteins could have clinical outcomes. We used a yeast-based expression assay to characterize human Rh variants resulting from non synonymous single nucleotide polymorphisms (nsSNPs with known or unknown clinical phenotypes and assessed their ammonium transport efficiency, protein level, localization and potential trans-dominant impact. The HsRhAG variants (I61R, F65S associated to overhydrated hereditary stomatocytosis (OHSt, a disease affecting erythrocytes, proved affected in intrinsic bidirectional ammonium transport. Moreover, this study reveals that the R202C variant of HsRhCG, the orthologue of mouse MmRhcg required for optimal urinary ammonium excretion and blood pH control, shows an impaired inherent ammonium transport activity. Urinary ammonium excretion was RHcg gene-dose dependent in mouse, highlighting MmRhcg as a limiting factor. HsRhCG(R202C may confer susceptibility to disorders leading to metabolic acidosis for instance. Finally, the analogous R211C mutation in the yeast ScMep2 homologue also impaired intrinsic activity consistent with a conserved functional role of the preserved arginine residue. The yeast expression assay used here constitutes an inexpensive, fast and easy tool to screen nsSNPs reported by high throughput sequencing or individual cases for functional alterations in Rh factors revealing potential causal variants.

  12. Functional Characterization of a Novel R2R3-MYB Transcription Factor Modulating the Flavonoid Biosynthetic Pathway from Epimedium sagittatum

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    Wenjun Huang

    2017-07-01

    Full Text Available Epimedium species have been widely used both as traditional Chinese medicinal plants and ornamental perennials. Both flavonols, acting as the major bioactive components (BCs and anthocyanins, predominantly contributing to the color diversity of Epimedium flowers belong to different classes of flavonoids. It is well-acknowledged that flavonoid biosynthetic pathway is predominantly regulated by R2R3-MYB transcription factor (TF as well as bHLH TF and WD40 protein at the transcriptional level. MYB TFs specifically regulating anthocyanin or flavonol biosynthetic pathway have been already isolated and functionally characterized from Epimedium sagittatum, but a R2R3-MYB TF involved in regulating both these two pathways has not been functionally characterized to date in Epimedium plants. In this study, we report the functional characterization of EsMYB9, a R2R3-MYB TF previously isolated from E. sagittatum. The previous study indicated that EsMYB9 belongs to a small subfamily of R2R3-MYB TFs containing grape VvMYB5a and VvMYB5b TFs, which regulate flavonoid biosynthetic pathway. The present studies show that overexpression of EsMYB9 in tobacco leads to increased transcript levels of flavonoid pathway genes and increased contents of anthocyanins and flavonols. Yeast two-hybrid assay indicates that the C-terminal region of EsMYB9 contributes to the autoactivation activity, and EsMYB9 interacts with EsTT8 or AtTT8 bHLH regulator. Transient reporter assay shows that EsMYB9 slightly activates the expression of EsCHS (chalcone synthase promoter in transiently transformed leaves of Nicotiana benthamiana, but the addition of AtTT8 or EsTT8 bHLH regulator strongly enhances the transcriptional activation of EsMYB9 against five promoters of the flavonoid pathway genes except EsFLS (flavonol synthase. In addition, co-transformation of EsMYB9 and EsTT8 in transiently transfected tobacco leaves strongly induces the expressions of flavonoid biosynthetic genes. The

  13. Genome-wide systematic characterization of the bZIP transcriptional factor family in tomato (Solanum lycopersicum L.).

    Science.gov (United States)

    Li, Dayong; Fu, Fuyou; Zhang, Huijuan; Song, Fengming

    2015-10-12

    might be involved in responses to various abiotic and biotic stresses as well as in response to light. This genome-wide systematic characterization identified a total of 69 members in the SlbZIP family and the analyses of the protein features and gene expression patterns provide useful clues for further functional characterization of the bZIP transcription factors in tomato.

  14. Cloning and Characterization of ThSHRs and ThSCR Transcription Factors in Taxodium Hybrid 'Zhongshanshan 406'.

    Science.gov (United States)

    Wang, Zhiquan; Yin, Yunlong; Hua, Jianfeng; Fan, Wencai; Yu, Chaoguang; Xuan, Lei; Yu, Fangyuan

    2017-07-20

    Among the GRAS family of transcription factors, SHORT ROOT (SHR) and SCARECROW (SCR) are key regulators of the formation of root tissues. In this study, we isolated and characterized two genes encoding SHR proteins and one gene encoding an SCR protein: ThSHR1 (Accession Number MF045148), ThSHR2 (Accession Number MF045149) and ThSCR (Accession Number MF045152) in the adventitious roots of Taxodium hybrid 'Zhongshanshan'. Gene structure analysis indicated that ThSHR1 , ThSHR2 and ThSCR are all intron free. Multiple protein sequence alignments showed that each of the corresponding proteins, ThSHR1, ThSHR2 and ThSCR, contained five well-conserved domains: leucine heptad repeat I (LHRI), the VHIID motif, leucine heptad repeat II (LHR II), the PFYRE motif, and the SAW motif. The phylogenetic analysis indicated that ThSCR was positioned in the SCR clade with the SCR proteins from eight other species, while ThSHR1 and ThSHR2 were positioned in the SHR clade with the SHR proteins from six other species. Temporal expression patterns of these genes were profiled during the process of adventitious root development on stem cuttings. Whereas expression of both ThSHR2 and ThSCR increased up to primary root formation before declining, that of ThSHR1 increased steadily throughout adventitious root formation. Subcellular localization studies in transgenic poplar protoplasts revealed that ThSHR1, ThSHR2 and ThSCR were localized in the nucleus. Collectively, these results suggest that the three genes encode Taxodium GRAS family transcription factors, and the findings contribute to improving our understanding of the expression and function of SHR and SCR during adventitious root production, which may then be manipulated to achieve high rates of asexual propagation of valuable tree species.

  15. Characterization of Factors Affecting Attainment of Glycemic Control in Asian Americans With Diabetes in a Culturally Specific Program

    Science.gov (United States)

    Le, Hung; Wong, Sophia; Iftikar, Tracy; Keenan, Hillary; King, George L.; Hsu, William C.

    2014-01-01

    Purpose The purpose of this study is to examine the effectiveness of a culturally specific pilot clinic for Asian Americans (AA) in reaching glycemic target and to characterize factors affecting the attainment of glycemic control in comparison with white counterparts. Methods This electronic health record review included all new AA patients with type 2 diabetes (n = 109) in a culturally specific program and a randomly selected sample of new white patients with type 2 diabetes (n = 218) in the adult clinic within the same time period and diabetes center. Results AA and whites had a comparable proportion of patients with A1C ≤7% (32.1%, 34.9%; P = .621) at baseline and after 12 months of care (48.6%, 56.0%; P = .210), with a similar A1C decline (−0.9% ± 1.6%, −0.8% ± 1.7%, P = .710) by 12 months. Factors associated with the lack of success in reaching target in AA but not in whites included older age, lower educational attainment, less likelihood of having health insurance, and a need for more educational visits. The percentage of AA reaching A1C ≤7%, as compared to whites, worsened among those with highest initial A1C when stratified by ascending quartiles (96.7% vs 85.2%, P = .101; 61.9% vs 58.9%, P = .813; 24.0% vs 37.7%, P = .230; 15.2% vs 35.4%, P = .044). Conclusion While a culturally specific diabetes program in a specialty setting achieved a similar glycemic outcome for AA compared with whites, reasons for not reaching glycemic target differed. The findings suggest that the elimination of diabetes disparities requires not only culturally and linguistically specific programs, but must also identify and address the socio-environmental differences unique to each population. PMID:23771841

  16. Characterization of a eukaryotic translation initiation factor 5A homolog from Tamarix androssowii involved in plant abiotic stress tolerance

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    Wang Liuqiang

    2012-07-01

    Full Text Available Abstract Background The eukaryotic translation initiation factor 5A (eIF5A promotes formation of the first peptide bond at the onset of protein synthesis. However, the function of eIF5A in plants is not well understood. Results In this study, we characterized the function of eIF5A (TaeIF5A1 from Tamarix androssowii. The promoter of TaeIF5A1 with 1,486 bp in length was isolated, and the cis-elements in the promoter were identified. A WRKY (TaWRKY and RAV (TaRAV protein can specifically bind to a W-box motif in the promoter of TaeIF5A1 and activate the expression of TaeIF5A1. Furthermore, TaeIF5A1, TaWRKY and TaRAV share very similar expression pattern and are all stress-responsive gene that functions in the abscisic acid (ABA signaling pathway, indicating that they are components of a single regulatory pathway. Transgenic yeast and poplar expressing TaeIF5A1 showed elevated protein levels combined with improved abiotic stresses tolerance. Furthermore, TaeIF5A1-transformed plants exhibited enhanced superoxide dismutase (SOD and peroxidase (POD activities, lower electrolyte leakage and higher chlorophyll content under salt stress. Conclusions These results suggested that TaeIF5A1 is involved in abiotic stress tolerance, and is likely regulated by transcription factors TaWRKY and TaRAV both of which can bind to the W-box motif. In addition, TaeIF5A1 may mediate stress tolerance by increasing protein synthesis, enhancing ROS scavenging by improving SOD and POD activities, and preventing chlorophyll loss and membrane damage. Therefore, eIF5A may play an important role in plant adaptation to changing environmental conditions.

  17. Characterization of molecular determinants of the conformational stability of macrophage migration inhibitory factor: leucine 46 hydrophobic pocket.

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    Farah El-Turk

    Full Text Available Macrophage Migration Inhibitory Factor (MIF is a key mediator of inflammatory responses and innate immunity and has been implicated in the pathogenesis of several inflammatory and autoimmune diseases. The oligomerization of MIF, more specifically trimer formation, is essential for its keto-enol tautomerase activity and probably mediates several of its interactions and biological activities, including its binding to its receptor CD74 and activation of certain signaling pathways. Therefore, understanding the molecular factors governing the oligomerization of MIF and the role of quaternary structure in modulating its structural stability and multifunctional properties is crucial for understanding the function of MIF in health and disease. Herein, we describe highly conserved intersubunit interactions involving the hydrophobic packing of the side chain of Leu46 onto the β-strand β3 of one monomer within a hydrophobic pocket from the adjacent monomer constituted by residues Arg11, Val14, Phe18, Leu19, Val39, His40, Val41, Val42, and Pro43. To elucidate the structural significance of these intersubunit interactions and their relative contribution to MIF's trimerization, structural stability and catalytic activity, we generated three point mutations where Leu46 was replaced by glycine (L46G, alanine (L46A and phenylalanine (L46F, and their structural properties, stability, oligomerization state, and catalytic activity were characterized using a battery of biophysical methods and X-ray crystallography. Our findings provide new insights into the role of the Leu46 hydrophobic pocket in stabilizing the conformational state of MIF in solution. Disrupting the Leu46 hydrophobic interaction perturbs the secondary and tertiary structure of the protein but has no effect on its oligomerization state.

  18. Characterization of a eukaryotic translation initiation factor 5A homolog from Tamarix androssowii involved in plant abiotic stress tolerance.

    Science.gov (United States)

    Wang, Liuqiang; Xu, Chenxi; Wang, Chao; Wang, Yucheng

    2012-07-26

    The eukaryotic translation initiation factor 5A (eIF5A) promotes formation of the first peptide bond at the onset of protein synthesis. However, the function of eIF5A in plants is not well understood. In this study, we characterized the function of eIF5A (TaeIF5A1) from Tamarix androssowii. The promoter of TaeIF5A1 with 1,486 bp in length was isolated, and the cis-elements in the promoter were identified. A WRKY (TaWRKY) and RAV (TaRAV) protein can specifically bind to a W-box motif in the promoter of TaeIF5A1 and activate the expression of TaeIF5A1. Furthermore, TaeIF5A1, TaWRKY and TaRAV share very similar expression pattern and are all stress-responsive gene that functions in the abscisic acid (ABA) signaling pathway, indicating that they are components of a single regulatory pathway. Transgenic yeast and poplar expressing TaeIF5A1 showed elevated protein levels combined with improved abiotic stresses tolerance. Furthermore, TaeIF5A1-transformed plants exhibited enhanced superoxide dismutase (SOD) and peroxidase (POD) activities, lower electrolyte leakage and higher chlorophyll content under salt stress. These results suggested that TaeIF5A1 is involved in abiotic stress tolerance, and is likely regulated by transcription factors TaWRKY and TaRAV both of which can bind to the W-box motif. In addition, TaeIF5A1 may mediate stress tolerance by increasing protein synthesis, enhancing ROS scavenging by improving SOD and POD activities, and preventing chlorophyll loss and membrane damage. Therefore, eIF5A may play an important role in plant adaptation to changing environmental conditions.

  19. Identification, functional characterization and expression pattern of myeloid differentiation factor 88 (MyD88) in Sepiella japonica.

    Science.gov (United States)

    Huo, Liping; Bao, Miaomiao; Lv, Zhenming; Chi, Changfeng; Wang, Tianming; Liu, Huihui

    2018-05-01

    Myeloid differentiation factor 88 (MyD88) is an adaptor protein involved in the interleukin-1 receptor and Toll-like receptor-induced activation of nuclear factor-κB (NF-κB). In this study a novel isoform of MyD88 in Sepiella japonica (SjMyD88) was cloned and functionally characterized (GenBank accession no. AQY56781.1). The complete cDNA sequence of SjMyD88 was 1912 bp and contained a 1017 bp open reading frame encoding 338 amino acid residues, which was similar to its mollusk orthologues in the length. BLASTp analysis suggested the deduced amino acids sequence of SjMyD88 shared high identity to the known MyD88, for instance, 64% identity with Octopus bimaculoides. Sequence analysis revealed two conserved domains, the N-terminal DD and the C-terminal TIR domain appeared in SjMyD88, which was consistent with MyD88 proteins from other species. The fusion expression of SjMyD88 and green fluorescent protein (EGFP) in HEK293 cells was conducted and cytoplasm localization was detected. Meanwhile, the TIR-pmCherry fusion protein showed red fluorescence and mainly distributed in the cytoplasm. After cotransfection MyD88-EGFP and TIR-pmCherry red obviously overlapped and changed to yellowish green. The results suggested that there was the interaction between homologous TIR-pmcherry and MyD88-EGFP. Tissues expression profiles analysis showed that SjMyD88 ubiquitously expressed in all tested tissues with the highest expression in the gills and livers except reproductive related tissue, and it was significantly induced in livers under LPS stress. These data provide insight into the roles of SjMyD88 in the TLR signaling pathway of S. japonica in response to pathogenic bacteria. Copyright © 2018. Published by Elsevier Ltd.

  20. Characterization of WRKY transcription factors in Solanum lycopersicum reveals collinearity and their expression patterns under cold treatment.

    Science.gov (United States)

    Chen, Lin; Yang, Yang; Liu, Can; Zheng, Yanyan; Xu, Mingshuang; Wu, Na; Sheng, Jiping; Shen, Lin

    2015-08-28

    WRKY transcription factors play an important role in cold defense of plants. However, little information is available about the cold-responsive WRKYs in tomato (Solanum lycopersicum). In the present study, a complete characterization of this gene family was described. Eighty WRKY genes in the tomato genome were identified. Almost all WRKY genes contain putative stress-responsive cis-elements in their promoter regions. Segmental duplications contributed significantly to the expansion of the SlWRKY gene family. Transcriptional analysis revealed notable differential expression in tomato tissues and expression patterns under cold stress, which indicated wide functional divergence in this family. Ten WRKYs in tomato were strongly induced more than 2-fold during cold stress. These genes represented candidate genes for future functional analysis of WRKYs involved in the cold-related signal pathways. Our data provide valuable information about tomato WRKY proteins and form a foundation for future studies of these proteins, especially for those that play an important role in response to cold stress. Copyright © 2015 Elsevier Inc. All rights reserved.

  1. Expression and characterization of insulin growth factor-I-enhanced green fluorescent protein fused protein as a tracer for immunoassay

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    Shi Ruina [College of Chemistry and Molecular Engineering, Peking University, Beijing 100871 (China); Huang Yong [College of Chemistry and Molecular Engineering, Peking University, Beijing 100871 (China); Wang Dan [College of Chemistry and Molecular Engineering, Peking University, Beijing 100871 (China); Zhao Meiping [College of Chemistry and Molecular Engineering, Peking University, Beijing 100871 (China); Li Yuanzong [College of Chemistry and Molecular Engineering, Peking University, Beijing 100871 (China)]. E-mail: yzli@pku.edu.cn

    2006-09-25

    The insulin-like growth factor-I (IGF-I) is an important polypeptide hormone under investigation for body metabolism study and for doping detection. Here, we describe for the first time the expression of a recombinant fusion protein of IGF-I and the enhanced green fluorescent protein (EGFP). The genetic fusion approach enables preparation of conjugates with 1:1 stoichiometry and homogeneous structure. The fused protein (EGFP-IGF-I) was expressed as a soluble protein in cytoplasm of Escherichia coli and its fluorescence and immunoreaction properties were thoroughly characterized. Finally, we demonstrated the utility of the EGFP-IGF-I fusion protein for the fluorescence immunoassay of IGF-1. The linear range of the assay is 1.6 x 10{sup -8} to 2.0 x 10{sup -6} M with a detection limit of 1.6 x 10{sup -8} M. To our knowledge, this is the first time that EGFP has been used as a quantitative label in a fusion protein to develop a quantitative assay for IGF-I. Furthermore, the use of genetically engineered fusion proteins, which combine peptide hormones with fluorescent protein, can lead to a new labeling approach to a number of bioanalytical applications.

  2. Expression and characterization of insulin growth factor-I-enhanced green fluorescent protein fused protein as a tracer for immunoassay

    International Nuclear Information System (INIS)

    Shi Ruina; Huang Yong; Wang Dan; Zhao Meiping; Li Yuanzong

    2006-01-01

    The insulin-like growth factor-I (IGF-I) is an important polypeptide hormone under investigation for body metabolism study and for doping detection. Here, we describe for the first time the expression of a recombinant fusion protein of IGF-I and the enhanced green fluorescent protein (EGFP). The genetic fusion approach enables preparation of conjugates with 1:1 stoichiometry and homogeneous structure. The fused protein (EGFP-IGF-I) was expressed as a soluble protein in cytoplasm of Escherichia coli and its fluorescence and immunoreaction properties were thoroughly characterized. Finally, we demonstrated the utility of the EGFP-IGF-I fusion protein for the fluorescence immunoassay of IGF-1. The linear range of the assay is 1.6 x 10 -8 to 2.0 x 10 -6 M with a detection limit of 1.6 x 10 -8 M. To our knowledge, this is the first time that EGFP has been used as a quantitative label in a fusion protein to develop a quantitative assay for IGF-I. Furthermore, the use of genetically engineered fusion proteins, which combine peptide hormones with fluorescent protein, can lead to a new labeling approach to a number of bioanalytical applications

  3. Characterization of a Staphylococcus aureus surface virulence factor that promotes resistance to oxidative killing and infectious endocarditis.

    Science.gov (United States)

    Malachowa, Natalia; Kohler, Petra L; Schlievert, Patrick M; Chuang, Olivia N; Dunny, Gary M; Kobayashi, Scott D; Miedzobrodzki, Jacek; Bohach, Gregory A; Seo, Keun Seok

    2011-01-01

    Staphylococcus aureus is a prominent human pathogen and a leading cause of community- and hospital-acquired bacterial infections worldwide. Herein, we describe the identification and characterization of the S. aureus 67.6-kDa hypothetical protein, named for the surface factor promoting resistance to oxidative killing (SOK) in this study. Sequence analysis showed that the SOK gene is conserved in all sequenced S. aureus strains and homologous to the myosin cross-reactive antigen of Streptococcus pyogenes. Immunoblotting and immunofluorescence analysis showed that SOK was copurified with membrane fractions and was exposed on the surface of S. aureus Newman and RN4220. Comparative analysis of wild-type S. aureus and an isogenic deletion strain indicated that SOK contributes to both resistance to killing by human neutrophils and to oxidative stress. In addition, the S. aureus sok deletion strain showed dramatically reduced aortic valve vegetation and bacterial cell number in a rabbit endocarditis model. These results, plus the suspected role of the streptococcal homologue in certain diseases such as acute rheumatic fever, suggest that SOK plays an important role in cardiovascular and other staphylococcal infections.

  4. Cloning, Identification, and Characterization of the rpoS-Like Sigma Factor rpoX from Vibrio alginolyticus

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    Jing-jing Zhao

    2009-01-01

    Full Text Available Vibrio alginolyticus ZJ-51 displays phase variation between opaque/rugose colonies (Op and translucent/smooth colonies (Tr. These colony variants show great differences in biofilm formation and motility. In this study, a gene encoding for an rpoS-like sigma factor, rpoX, has been cloned and characterized. The absence of rpoX did not affect colony switching rate but did decrease biofilm formation in both the Op and the Tr variants. When challenged with hydrogen peroxide, the ΔrpoX in the Op background showed a slightly higher survival rate compared with the wild type, whereas survival was decreased in the Tr background. Deletion of rpoX in the Tr background resulted in a higher ability to resist ethanol challenges and to survive hyperosmolarity challenges, and in the Op background the opposite phenotype was observed. This indicates that the rpoX gene is involved in biofilm formation and stress response but the effects are controlled by colony phase variation in V. alginolyticus.

  5. High-throughput proteomic characterization of plasma rich in growth factors (PRGF-Endoret)-derived fibrin clot interactome.

    Science.gov (United States)

    Anitua, Eduardo; Prado, Roberto; Azkargorta, Mikel; Rodriguez-Suárez, Eva; Iloro, Ibon; Casado-Vela, Juan; Elortza, Felix; Orive, Gorka

    2015-11-01

    Plasma rich in growth factors (PRGF®-Endoret®) is an autologous technology that contains a set of proteins specifically addressed to wound healing and tissue regeneration. The scaffold formed by using this technology is a clot mainly composed of fibrin protein, forming a three-dimensional (3D) macroscopic network. This biomaterial is easily obtained by biotechnological means from blood and can be used in a range of situations to help wound healing and tissue regeneration. Although the main constituent of this clot is the fibrin scaffold, little is known about other proteins interacting in this clot that may act as adjuvants in the healing process. The aim of this study was to characterize the proteins enclosed by PRGF-Endoret scaffold, using a double-proteomic approach that combines 1D-SDS-PAGE approach followed by LC-MS/MS, and 2-DE followed by MALDI-TOF/TOF. The results presented here provide a description of the catalogue of key proteins in close contact with the fibrin scaffold. The obtained lists of proteins were grouped into families and networks according to gene ontology. Taken together, an enrichment of both proteins and protein families specifically involved in tissue regeneration and wound healing has been found. Copyright © 2013 John Wiley & Sons, Ltd.

  6. Lobular carcinoma in situ and invasive lobular breast cancer are characterized by enhanced expression of transcription factor AP-2β.

    Science.gov (United States)

    Raap, Mieke; Gronewold, Malte; Christgen, Henriette; Glage, Silke; Bentires-Alj, Mohammad; Koren, Shany; Derksen, Patrick W; Boelens, Mirjam; Jonkers, Jos; Lehmann, Ulrich; Feuerhake, Friedrich; Kuehnle, Elna; Gluz, Oleg; Kates, Ronald; Nitz, Ulrike; Harbeck, Nadia; Kreipe, Hans H; Christgen, Matthias

    2018-01-01

    Transcription factor AP-2β (TFAP2B) regulates embryonic organ development and is overexpressed in alveolar rhabdomyosarcoma, a rare childhood malignancy. Gene expression profiling has implicated AP-2β in breast cancer (BC). This study characterizes AP-2β expression in the mammary gland and in BC. AP-2β protein expression was assessed in the normal mammary gland epithelium, in various reactive, metaplastic and pre-invasive neoplastic lesions and in two clinical BC cohorts comprising >2000 patients. BCs from various genetically engineered mouse (GEM) models were also evaluated. Human BC cell lines served as functional models to study siRNA-mediated inhibition of AP-2β. The normal mammary gland epithelium showed scattered AP-2β-positive cells in the luminal cell layer. Various reactive and pre-invasive neoplastic lesions, including apocrine metaplasia, usual ductal hyperplasia and lobular carcinoma in situ (LCIS) showed enhanced AP-2β expression. Cases of ductal carcinoma in situ (DCIS) were more often AP-2β-negative (Pinvasive BC cohorts, AP-2β-positivity was associated with the lobular BC subtype (Plobular BC cell lines in vitro. In summary, AP-2β is a new mammary epithelial differentiation marker. Its expression is preferentially retained and enhanced in LCIS and invasive lobular BC and has prognostic implications. Our findings indicate that AP-2β controls tumor cell proliferation in this slow-growing BC subtype.

  7. Characterization of capsaicin synthase and identification of its gene (csy1) for pungency factor capsaicin in pepper (Capsicum sp.)

    Science.gov (United States)

    Prasad, B. C. Narasimha; Kumar, Vinod; Gururaj, H. B.; Parimalan, R.; Giridhar, P.; Ravishankar, G. A.

    2006-01-01

    Capsaicin is a unique alkaloid of the plant kingdom restricted to the genus Capsicum. Capsaicin is the pungency factor, a bioactive molecule of food and of medicinal importance. Capsaicin is useful as a counterirritant, antiarthritic, analgesic, antioxidant, and anticancer agent. Capsaicin biosynthesis involves condensation of vanillylamine and 8-methyl nonenoic acid, brought about by capsaicin synthase (CS). We found that CS activity correlated with genotype-specific capsaicin levels. We purified and characterized CS (≈35 kDa). Immunolocalization studies confirmed that CS is specifically localized to the placental tissues of Capsicum fruits. Western blot analysis revealed concomitant enhancement of CS levels and capsaicin accumulation during fruit development. We determined the N-terminal amino acid sequence of purified CS, cloned the CS gene (csy1) and sequenced full-length cDNA (981 bp). The deduced amino acid sequence of CS from full-length cDNA was 38 kDa. Functionality of csy1 through heterologous expression in recombinant Escherichia coli was also demonstrated. Here we report the gene responsible for capsaicin biosynthesis, which is unique to Capsicum spp. With this information on the CS gene, speculation on the gene for pungency is unequivocally resolved. Our findings have implications in the regulation of capsaicin levels in Capsicum genotypes. PMID:16938870

  8. Preparation of Gc protein-derived macrophage activating factor (GcMAF) and its structural characterization and biological activities.

    Science.gov (United States)

    Mohamad, Saharuddin Bin; Nagasawa, Hideko; Uto, Yoshihiro; Hori, Hitoshi

    2002-01-01

    Gc protein has been reported to be a precursor of Gc protein-derived macrophage activation factor (GcMAF) in the inflammation-primed macrophage activation cascade. An inducible beta-galactosidase of B cells and neuraminidase of T cells convert Gc protein to GcMAF. Gc protein from human serum was purified using 25(OH)D3 affinity column chromatography and modified to GcMAF using immobilized glycosidases (beta-galactosidase and neuraminidase) The sugar moiety structure of GcMAF was characterized by lectin blotting by Helix pomatia agglutinin. The biological activities of GcMAF were evaluated by a superoxide generation assay and a phagocytosis assay. We successfully purified Gc protein from human serum. GcMAF was detected by lectin blotting and showed a high biological activity. Our results support the importance of the terminal N-acetylgalactosamine moiety in the GcMAF-mediated macrophage activation cascade, and the existence of constitutive GcMAF in human serum. These preliminary data are important for designing small molecular GcMAF mimics.

  9. Characterization of transcription factor networks involved in umbilical cord blood CD34+ stem cells-derived erythropoiesis.

    Directory of Open Access Journals (Sweden)

    Biaoru Li

    Full Text Available Fetal stem cells isolated from umbilical cord blood (UCB possess a great capacity for proliferation and differentiation and serve as a valuable model system to study gene regulation. Expanded knowledge of the molecular control of hemoglobin synthesis will provide a basis for rational design of therapies for β-hemoglobinopathies. Transcriptome data are available for erythroid progenitors derived from adult stem cells, however studies to define molecular mechanisms controlling globin gene regulation during fetal erythropoiesis are limited. Here, we utilize UCB-CD34+ stem cells induced to undergo erythroid differentiation to characterize the transcriptome and transcription factor networks (TFNs associated with the γ/β-globin switch during fetal erythropoiesis. UCB-CD34+ stem cells grown in the one-phase liquid culture system displayed a higher proliferative capacity than adult CD34+ stem cells. The γ/β-globin switch was observed after day 42 during fetal erythropoiesis in contrast to adult progenitors where the switch occurred around day 21. To gain insights into transcription factors involved in globin gene regulation, microarray analysis was performed on RNA isolated from UCB-CD34+ cell-derived erythroid progenitors harvested on day 21, 42, 49 and 56 using the HumanHT-12 Expression BeadChip. After data normalization, Gene Set Enrichment Analysis identified transcription factors (TFs with significant changes in expression during the γ/β-globin switch. Forty-five TFs were silenced by day 56 (Profile-1 and 30 TFs were activated by day 56 (Profile-2. Both GSEA datasets were analyzed using the MIMI Cytoscape platform, which discovered TFNs centered on KLF4 and GATA2 (Profile-1 and KLF1 and GATA1 for Profile-2 genes. Subsequent shRNA studies in KU812 leukemia cells and human erythroid progenitors generated from UCB-CD34+ cells supported a negative role of MAFB in γ-globin regulation. The characteristics of erythroblasts derived from UCB-CD34

  10. Comprehensive characterization and RNA-Seq profiling of the HD-Zip transcription factor family in soybean (Glycine max) during dehydration and salt stress

    Science.gov (United States)

    The homeodomain leucine zipper (HD-Zip) transcription factor family is one of the largest plant specific superfamilies, and includes genes with roles in modulation of plant growth and response to environmental stresses. Many HD-Zip genes are well characterized in Arabidopsis (Arabidopsis thaliana), ...

  11. Functional Characterization of NAC and MYB Transcription Factors Involved in Regulation of Biomass Production in Switchgrass (Panicum virgatum.

    Directory of Open Access Journals (Sweden)

    Ruiqin Zhong

    Full Text Available Switchgrass is a promising biofuel feedstock due to its high biomass production and low agronomic input requirements. Because the bulk of switchgrass biomass used for biofuel production is lignocellulosic secondary walls, studies on secondary wall biosynthesis and its transcriptional regulation are imperative for designing strategies for genetic improvement of biomass production in switchgrass. Here, we report the identification and functional characterization of a group of switchgrass transcription factors, including several NACs (PvSWNs and a MYB (PvMYB46A, for their involvement in regulating secondary wall biosynthesis. PvSWNs and PvMYB46A were found to be highly expressed in stems and their expression was closely associated with sclerenchyma cells. Overexpression of PvSWNs and PvMYB46A in Arabidopsis was shown to result in activation of the biosynthetic genes for cellulose, xylan and lignin and ectopic deposition of secondary walls in normally parenchymatous cells. Transactivation and complementation studies demonstrated that PvSWNs were able to activate the SNBE-driven GUS reporter gene and effectively rescue the secondary wall defects in the Arabidopsis snd1 nst1 double mutant, indicating that they are functional orthologs of Arabidopsis SWNs. Furthermore, we showed that PvMYB46A could activate the SMRE-driven GUS reporter gene and complement the Arabidopsis myb46 myb83 double mutant, suggesting that it is a functional ortholog of Arabidopsis MYB46/MYB83. Together, these results indicate that PvSWNs and PvMYB46A are transcriptional switches involved in regulating secondary wall biosynthesis, which provides molecular tools for genetic manipulation of biomass production in switchgrass.

  12. Characterization and molecular features of the cell surface receptor for human granulocyte-macrophage colony-stimulating factor

    International Nuclear Information System (INIS)

    Chiba, S.; Tojo, A.; Kitamura, T.; Urabe, A.; Miyazono, K.; Takaku, F.

    1990-01-01

    The receptors for human granulocyte-macrophage colony-stimulating factor (GM-CSF) on the surfaces of normal and leukemic myeloid cells were characterized using 125I-labeled bacterially synthesized GM-CSF. The binding was rapid, specific, time dependent, and saturable. Scatchard analysis of the 125I-GM-CSF binding to peripheral blood neutrophils indicated the presence of a single class of binding site (Kd = 99 +/- 21 pM; 2,304 +/- 953 sites/cell). However, for peripheral blood monocytes and two GM-CSF-responsive myeloid cell lines (U-937 and TF-1), the Scatchard plots were biphasic curvilinear, which were best fit by curves derived from two binding site model: one with high affinity (Kd1 = 10-40 pM) and the other with low affinity (Kd2 = 0.9-2.0 nM). For U-937 cells, the number of high-affinity receptors was 1,058 +/- 402 sites/cell and that of low-affinity receptors was estimated to be 10,834 +/- 2,396 sites/cell. Cross-linking studies yielded three major bands with molecular masses of 150 kDa, 115 kDa, and 95 kDa, which were displaced by an excess amount of unlabeled GM-CSF, suggesting 135-kDa, 100-kDa, and 80-kDa species for the individual components of the human GM-CSF receptor. These bands comigrated for different cell types including peripheral blood neutrophils, U-937 cells and TF-1 cells. In experiments using U-937 cells, only the latter two bands appeared to be labeled in a dose-dependent manner in a low-affinity state. These results suggest that the human GM-CSF receptor possibly forms a multichain complex

  13. Fontan-associated protein-losing enteropathy and plastic bronchitis: characterizing current-era risk factors, course, and progression

    Science.gov (United States)

    Schumacher, Kurt R.; Stringer, Kathleen A.; Donohue, Janet E.; Yu, Sunkyung; Shaver, Ashley; Caruthers, Regine L.; Zikmund-Fisher, Brian J.; Fifer, Carlen; Goldberg, Caren; Russell, Mark W.

    2015-01-01

    Objective Characterize the medical history, disease progression, and treatment of current-era patients with the rare diseases Fontan-associated protein losing enteropathy (PLE) and plastic bronchitis (PB). Study Design A novel survey that queried demographics, medical details, and treatment information was piloted and placed online via a Facebook portal allowing social media to power the study. Participation regardless of PLE or PB diagnosis was allowed. Case control analyses compared patients with PLE and PB to uncomplicated control Fontan patients. Results The survey was completed by 671 subjects including 76 with PLE, 46 with PB, and 7 with both. Median PLE diagnosis was 2.5 years post-Fontan. Hospitalization for PLE occurred in 71% with 41% hospitalized ≥ 3 times. Therapy varied significantly. PLE patients more commonly had hypoplastic left ventricle (62% vs 44% control; OR 2.8, 95% CI 1.4–5.5), chylothorax (66% vs 41%; OR 3.0, CI 1.6–5.3), and cardiothoracic surgery in addition to staged palliation (17% vs 5%; OR 4.3, CI 1.6, 11.2). Median PB diagnosis was 2 years post-Fontan. Hospitalization for PB occurred in 91% with 61% hospitalized ≥3 times. Therapy was very diverse. PB patients more commonly had chylothorax at any surgery (72% vs 51%; OR 2.5, CI 1.2–5.1) and seasonal allergies (52% vs 36%; OR 2.0, CI 1.0–3.9). Conclusions Patient-specific factors are associated with diagnoses of PLE or PB. Treatment strategies are diverse without clear patterns. These results provide a foundation upon which to design future therapeutic studies and identify a clear need for forming consensus approaches to treatment. PMID:25661406

  14. Identification and molecular characterization of MYB Transcription Factor Superfamily in C4 model plant foxtail millet (Setaria italica L.).

    Science.gov (United States)

    Muthamilarasan, Mehanathan; Khandelwal, Rohit; Yadav, Chandra Bhan; Bonthala, Venkata Suresh; Khan, Yusuf; Prasad, Manoj

    2014-01-01

    MYB proteins represent one of the largest transcription factor families in plants, playing important roles in diverse developmental and stress-responsive processes. Considering its significance, several genome-wide analyses have been conducted in almost all land plants except foxtail millet. Foxtail millet (Setaria italica L.) is a model crop for investigating systems biology of millets and bioenergy grasses. Further, the crop is also known for its potential abiotic stress-tolerance. In this context, a comprehensive genome-wide survey was conducted and 209 MYB protein-encoding genes were identified in foxtail millet. All 209 S. italica MYB (SiMYB) genes were physically mapped onto nine chromosomes of foxtail millet. Gene duplication study showed that segmental- and tandem-duplication have occurred in genome resulting in expansion of this gene family. The protein domain investigation classified SiMYB proteins into three classes according to number of MYB repeats present. The phylogenetic analysis categorized SiMYBs into ten groups (I-X). SiMYB-based comparative mapping revealed a maximum orthology between foxtail millet and sorghum, followed by maize, rice and Brachypodium. Heat map analysis showed tissue-specific expression pattern of predominant SiMYB genes. Expression profiling of candidate MYB genes against abiotic stresses and hormone treatments using qRT-PCR revealed specific and/or overlapping expression patterns of SiMYBs. Taken together, the present study provides a foundation for evolutionary and functional characterization of MYB TFs in foxtail millet to dissect their functions in response to environmental stimuli.

  15. Identification and molecular characterization of MYB Transcription Factor Superfamily in C4 model plant foxtail millet (Setaria italica L..

    Directory of Open Access Journals (Sweden)

    Mehanathan Muthamilarasan

    Full Text Available MYB proteins represent one of the largest transcription factor families in plants, playing important roles in diverse developmental and stress-responsive processes. Considering its significance, several genome-wide analyses have been conducted in almost all land plants except foxtail millet. Foxtail millet (Setaria italica L. is a model crop for investigating systems biology of millets and bioenergy grasses. Further, the crop is also known for its potential abiotic stress-tolerance. In this context, a comprehensive genome-wide survey was conducted and 209 MYB protein-encoding genes were identified in foxtail millet. All 209 S. italica MYB (SiMYB genes were physically mapped onto nine chromosomes of foxtail millet. Gene duplication study showed that segmental- and tandem-duplication have occurred in genome resulting in expansion of this gene family. The protein domain investigation classified SiMYB proteins into three classes according to number of MYB repeats present. The phylogenetic analysis categorized SiMYBs into ten groups (I-X. SiMYB-based comparative mapping revealed a maximum orthology between foxtail millet and sorghum, followed by maize, rice and Brachypodium. Heat map analysis showed tissue-specific expression pattern of predominant SiMYB genes. Expression profiling of candidate MYB genes against abiotic stresses and hormone treatments using qRT-PCR revealed specific and/or overlapping expression patterns of SiMYBs. Taken together, the present study provides a foundation for evolutionary and functional characterization of MYB TFs in foxtail millet to dissect their functions in response to environmental stimuli.

  16. Functional characterization of viral tumor necrosis factor receptors encoded by cyprinid herpesvirus 3 (CyHV3) genome.

    Science.gov (United States)

    Yi, Yang; Qi, Hemei; Yuan, Jimin; Wang, Rui; Weng, Shaoping; He, Jianguo; Dong, Chuanfu

    2015-08-01

    Cyprinid herpesvirus 3 (CyHV3) is a large double-stranded DNA virus of Alloherpesviridae family in the order Herpesvirales. It causes significant morbidity and mortality in common carp and its ornamental koi variety, and threatens the aquaculture industries worldwide. Mimicry of cytokines and cytokine receptors is a particular strategy for large DNA viruses in modulating the host immune response. Here, we report the identification and characterization of two novel viral homologues of tumor necrosis factor receptor (TNFR) encoded by CyHV3-ORF4 and -ORF12, respectively. CyHV3-ORF4 was identified as a homologue of HVEM and CyHV3-ORF12 as a homologue of TNFRSF1. Overexpression of ORF4 and ORF12 in zebrafish embryos results in embryonic lethality, morphological defects and increased apoptosis. Although we failed to identify any interaction between the two vTNFRs and their potential ligands in zebrafish TNF superfamily by yeast two-hybrid system, the expression of some genes in TNF superfamily or TNFR superfamily were mis-regulated in ORF4 or ORF12-overexpressing embryos, especially the death receptor zHDR and its cognate ligand DL1b. Further studies showed that the apoptosis induced by the both CyHV3 vTNFRs is mainly activated through the intrinsic apoptotic pathway and requires the crosstalk between the intrinsic and extrinsic apoptotic pathway. Additionally, using RT-qPCR and Western blot assays, the expression patterns of the both vTNFRs were also analyzed during CyHV3 productive infection. Collectively, this is the first functional study of two unique vTNFRs encoded by a herpesvirus infecting non-mammalian vertebrates, which may provide novel insights into viral immune regulation mechanism and the pathogenesis of CyHV3 infection. Copyright © 2015 Elsevier Ltd. All rights reserved.

  17. Systematic in vitro and in vivo characterization of Leukemia-inhibiting factor- and Fibroblast growth factor-derived porcine induced pluripotent stem cells

    DEFF Research Database (Denmark)

    Secher, Jan Ole Bertelsen; Ceylan, Ahmet; Mazzoni, Gianluca

    2017-01-01

    Derivation and stable maintenance of porcine induced pluripotent stem cells (piPSCs) is challenging. We herein systematically analyzed two piPSC lines, derived by lentiviral transduction and cultured under either leukemia inhibitory factor (LIF) or fibroblast growth factor (FGF) conditions, to sh...

  18. Analysis of WRKY transcription factors and characterization of two Botrytis cinerea-responsive LrWRKY genes from Lilium regale.

    Science.gov (United States)

    Cui, Qi; Yan, Xiao; Gao, Xue; Zhang, Dong-Mei; He, Heng-Bin; Jia, Gui-Xia

    2018-06-01

    A major constraint in producing lilies is gray mold caused by Botrytis elliptica and B. cinerea. WRKY transcription factors play important roles in plant immune responses. However, limited information is available about the WRKY gene family in lily plants. In this study, 23 LrWRKY genes with complete WRKY domains were identified from the Botrytis-resistant species Lilium regale. The putative WRKY genes were divided into seven subgroups (Group I, IIa-e, and III) according to their structural features. Sequence alignment revealed that LrWRKY proteins have a highly conserved WRKYGQK domain and a variant, the WRKYGKK domain, and these proteins generally contained similar motif compositions throughout the same subgroup. Functional annotation predicted they might be involved in biological processes related to abiotic and biotic stresses. A qRT-PCR analysis confirmed that expression of six LrWRKY genes in L. regale or the susceptible Asian hybrid 'Yale' was induced by B. cinerea infection. Among these genes, LrWRKY4, LrWRKY8 and LrWRKY10 were expressed at a higher level in L. regale than 'Yale', while the expression of LrWRKY6 and LrWRKY12 was lower in L. regale. Furthermore, LrWRKY4 and LrWRKY12 genes, which also respond to salicylic acid (SA) and methyl jasmonate (MeJA) treatments, were isolated from L. regale. Subcellular localization analysis determined that they were targeted to the nucleus. Constitutive expression of LrWRKY4 and LrWRKY12 in Arabidopsis resulted in plants that were more resistant to B. cinerea than wild-type plants. This resistance was coupled with the transcriptional changes of SA and JA-responsive genes. Overall, our study provides valuable information about the structural and functional characterization of LrWRKY genes that will not only deepen our understanding of the molecular mechanisms underlying the defense of lily against B. cinerea but also offer potential targets for cultivar improvement via biotechnology. Copyright © 2018 Elsevier Masson

  19. Molecular characterization of banana NAC transcription factors and their interactions with ethylene signalling component EIL during fruit ripening.

    Science.gov (United States)

    Shan, Wei; Kuang, Jian-fei; Chen, Lei; Xie, Hui; Peng, Huan-huan; Xiao, Yun-yi; Li, Xue-ping; Chen, Wei-xin; He, Quan-guang; Chen, Jian-ye; Lu, Wang-jin

    2012-09-01

    The plant-specific NAC (NAM, ATAF1/2, and CUC2) transcription factors (TFs) play important roles in plant growth, development, and stress responses. However, the precise role of NAC TFs in relation to fruit ripening is poorly understood. In this study, six NAC genes, designated MaNAC1-MaNAC6, were isolated and characterized from banana fruit. Subcellular localization showed that MaNAC1-MaNAC5 proteins localized preferentially to the nucleus, while MaNAC6 was distributed throughout the entire cell. A transactivation assay in yeast demonstrated that MaNAC4 and MaNAC6, as well as their C-terminal regions, possessed trans-activation activity. Gene expression profiles in fruit with four different ripening characteristics, including natural, ethylene-induced, 1-methylcyclopropene (1-MCP)-delayed, and a combination of 1-MCP with ethylene treatment, revealed that the MaNAC genes were differentially expressed in peel and pulp during post-harvest ripening. MaNAC1 and MaNAC2 were apparently upregulated by ethylene in peel and pulp, consistent with the increase in ethylene production. In contrast, MaNAC3 in peel and pulp and MaNAC5 in peel were constitutively expressed, and transcripts of MaNAC4 in peel and pulp and MaNAC6 in peel decreased, while MaNAC5 or MaNAC6 in pulp increased slightly during fruit ripening. Furthermore, the MaNAC2 promoter was activated after ethylene application, further enhancing the involvement of MaNAC2 in fruit ripening. More importantly, yeast two-hybrid and bimolecular fluorescence complementation analyses confirmed that MaNAC1/2 physically interacted with a downstream component of ethylene signalling, ethylene insensitive 3 (EIN3)-like protein, termed MaEIL5, which was downregulated during ripening. Taken together, these results suggest that MaNACs such as MaNAC1/MaNAC2, may be involved in banana fruit ripening via interaction with ethylene signalling components.

  20. Characterization of fibroblast growth factor receptor 2 overexpression in the human breast cancer cell line SUM-52PE

    International Nuclear Information System (INIS)

    Tannheimer, Stacey L; Rehemtulla, Alnawaz; Ethier, Stephen P

    2000-01-01

    three times more transformed foci in soft agar than C1 variants (both IIIb), whereas full length FGFR2 and FGFR1 (both IIIc variants) showed no transforming activity [4]. Previous studies [5,6] have found amplification and overexpression of FGFR2 in 5-10% of primary breast cancer specimens. A recent study [7] done using a tissue array consisting of 372 primary breast cancer specimens found a 5% incidence of FGFR2 amplification. To our knowledge, none of the HBC cell lines studied thus far have an FGFR2 gene amplification, although overexpression of FGFR2 message and protein has been documented for some breast cancer cell lines [6,8,9]. SUM-52PE is a breast cancer cell line previously isolated in our laboratory that grows under serum-free and epidermal growth factor-free conditions, has high levels of tyrosine-phosphorylated membrane proteins, and has the capacity to invade and grow under anchorage-independent conditions [10,11,12]. This cell line exhibits all of the important hallmarks of transformed, highly malignant cells. Therefore, SUM-52PE was used as a model to study the diversity of FGFR2 expression in a breast cancer cell line that has true amplification and overexpression of FGFR2. This study was conducted to examine the degree of FGFR2 amplification and overexpression in the breast cancer cell line SUM-52PE. Subsequent sequencing and characterization of individual FGFR2 variants cloned from the SUM-52PE cell line was completed to determine the complexity of FGFR2 alternative splicing in the context of a highly metastatic breast cancer cell line. Southern, Northern and Western blot analyses were done in order to determine the degree of FGFR2 amplification and overexpression in the breast cancer cell line SUM-52PE. Individual FGFR2 variants were cloned out of SUM-52PE using FGFR2-specific primers in a reverse transcription (RT) polymerase chain reaction (PCR). FGFR2 cDNAs were characterized by restriction fragment analysis, sequencing and transient transfection

  1. Characterization of the factors having an influence on the evolution of the EPR signal of irradiated alanine

    International Nuclear Information System (INIS)

    Feaugas-Le-Berre, Valerie

    1999-01-01

    EPR/alanine dosimetry has been used by the LNHB (Laboratoire National Henri Becquerel) since many years for applied metrology. This technic is based on the measurement of the EPR signal of the free radicals induced in alanine by irradiation. The aim of this work is to characterize the factors having an influence on the evolution of the amplitude of the EPR signal of irradiated alanine to limit the uncertainties on the determination of the absorbed dose. The first step of this work has been the choice of the dosimeter. A bibliographic study completed by experiments on the response of alanine isomers to the dose and on its stability with time has lead us to choose L-α-alanine powder as dosimeter. The influence of the recording parameter of the spectrometer on the characteristics of the EPR spectrum has then been studied. This has enabled us to optimize the recording conditions of EPR spectra. As the angular anisotropy of the EPR signal limits the measurements reproducibility, an experimental protocol has been defined to solve this problem. The repeatability of the measurements has been enhanced by modifying the spectrometer and using an internal standard constituted of single crystals of CuSO 4 .5H 2 O. As the amplitude of the EPR signal is sensitive to the measurement temperature, a method of normalization of the results to 20 C has been determined. We have studied the influence of an irradiation parameter and of environmental parameters. We have shown that the EPR signal amplitude increases with irradiation temperature. The EPR signal amplitude and its evolution vary strongly with storage conditions (temperature and moisture) of the dosimeter before and after irradiation. The presence of moisture in alanine powder leads to a loss of signal amplitude. The dosimeters exposition to light also entails a loss of amplitude. Oxygen does not influence the EPR spectrum of alanine. We have noticed that the EPR signal amplitude of samples stored in absence of moisture

  2. Characterization and Evaluation of 600 V Range Devices for Active Power Factor Correction in Boundary and Continuous Conduction Modes

    DEFF Research Database (Denmark)

    Hernandez Botella, Juan Carlos; Petersen, Lars Press; Andersen, Michael A. E.

    2015-01-01

    Traditional characterization of semiconductors switching dynamics is performed based on clamped inductive load measurements using the double pulse tester (DPT) configuration. This approach is valid for converters operating in continuous conduction mode (CCM), however in boundary conduction mode...

  3. An In Vivo Characterization of Trophic Factor Production Following Neural Precursor Cell or Bone Marrow Stromal Cell Transplantation for Spinal Cord Injury

    Science.gov (United States)

    Hawryluk, Gregory W.J.; Mothe, Andrea; Wang, Jian; Wang, Shelly; Tator, Charles

    2012-01-01

    Cellular transplantation strategies for repairing the injured spinal cord have shown consistent benefit in preclinical models, and human clinical trials have begun. Interactions between transplanted cells and host tissue remain poorly understood. Trophic factor secretion is postulated a primary or supplementary mechanism of action for many transplanted cells, however, there is little direct evidence to support trophin production by transplanted cells in situ. In the present study, trophic factor expression was characterized in uninjured, injured-untreated, injured-treated with transplanted cells, and corresponding control tissue from the adult rat spinal cord. Candidate trophic factors were identified in a literature search, and primers were designed for these genes. We examined in vivo trophin expression in 3 paradigms involving transplantation of either brain or spinal cord-derived neural precursor cells (NPCs) or bone marrow stromal cells (BMSCs). Injury without further treatment led to a significant elevation of nerve growth factor (NGF), leukemia inhibitory factor (LIF), insulin-like growth factor-1 (IGF-1), and transforming growth factor-β1 (TGF-β1), and lower expression of vascular endothelial growth factor isoform A (VEGF-A) and platelet-derived growth factor-A (PDGF-A). Transplantation of NPCs led to modest changes in trophin expression, and the co-administration of intrathecal trophins resulted in significant elevation of the neurotrophins, glial-derived neurotrophic factor (GDNF), LIF, and basic fibroblast growth factor (bFGF). BMSCs transplantation upregulated NGF, LIF, and IGF-1. NPCs isolated after transplantation into the injured spinal cord expressed the neurotrophins, ciliary neurotrophic factor (CNTF), epidermal growth factor (EGF), and bFGF at higher levels than host cord. These data show that trophin expression in the spinal cord is influenced by injury and cell transplantation, particularly when combined with intrathecal trophin infusion

  4. Upper Paleozoic mafic and intermediate volcanic rocks of the Mount Pleasant caldera associated with the Sn-W deposit in southwestern New Brunswick (Canada): Petrogenesis and metallogenic implications

    Science.gov (United States)

    Dostal, Jaroslav; Jutras, Pierre

    2016-10-01

    crystallization and evolved along a tholeiitic trend of "early iron" enrichment (non-oxic conditions). The contaminated magma of the second association followed a calc-alkaline fractionation trend of "no iron" enrichment (oxidizing conditions) characterized by a high PH2O and PO2 environment at the mid-crust levels. The Piskahegan Group, which is associated with an important polymetallic mineral deposit, differs from the numerous non-mineralized rift-related volcanic suites of the regional Upper Devonian to Lower Carboniferous successions in the Maritimes Basin by the presence of a significant amount of coeval calc-alkaline andesite, which may be an indicator of potential mineralization.

  5. Expression and characterization of recombinant human factor V and a mutant lacking a major portion of the connecting region

    International Nuclear Information System (INIS)

    Kane, W.H.; Devore-Carter, D.; Ortel, T.L.

    1990-01-01

    Human coagulation factor V is a protein cofactor that is an essential component of the prothrombinase complex. A full-length factor V cDNA has been subcloned into the mammalian expression vector pDX and used to transfect COS cells. Approximately 95 ± 4% of the recombinant human factor V (rHFV) synthesized in COS cells is secreted into the culture medium. Factor V activity determined by fibrometer assay increased approximately 5-fold from 0.027 ± 0.012 to 0.124 ± 0.044 unit/mL following activation by the factor V activating enzyme from Russell's viper venom (RVV-V). A chromogenic assay specific for factor Va indicated that recombinant factor V had 3.8 ± 1.3% of the activity of the activated protein. The estimated specific activity of the recombinant factor Va was approximately 1,800 ± 500 units/mg, which is similar to the specific activity of purified plasma factor Va of 1,700-2,000 units/mg. Immunoprecipitation of [ 35 S]methionine-labeled rHFV revealed a single high molecular mass component. Treatment of rHFV with thrombin or RVV-V resulted in the formation of proteolytic products that were similar to those seen with plasma factor V. The authors have also expressed a mutant, rHFV-des-B 811-1441 , that lacks a large portion of the highly glycosylated connecting region that is present in factor V. This mutant constitutively expressed 38 ± 7% of the activity of the RVV-V-activated protein. These results suggest that one of the functions of the large connecting region in factor V is to inhibit constitutive procoagulant activity

  6. Characterization of two homeodomain transcription factors with critical but distinct roles in virulence in the vascular pathogen Verticillium dahliae

    Science.gov (United States)

    Vascular wilt caused by Verticillium dahliae is a destructive disease that represents a chronic economic problem for crop production worldwide. In this work, we characterized two new regulators of pathogenicity in this species. Vph1 (VDAG_06555) was identified in a candidate gene approach as a putat...

  7. Characterization of the domains of E. coli initiation factor IF2 responsible for recognition of the ribosome

    DEFF Research Database (Denmark)

    Manuel Palacios Moreno, Juan; Andersen, Lars Dyrskjøt; Egebjerg Kristensen, Janni

    1999-01-01

    We have studied the interactions between the ribosome and the domains of Escherichia coli translation initiation factor 2, using an in vitro ribosomal binding assay with wild-type forms, N- and C-terminal truncated forms of IF2 as well as isolated structural domains. A deletion mutant of the factor...

  8. Characterization of five ECF sigma factors in the genome of Pseudomonas syringae pv. syringae B728a.

    Directory of Open Access Journals (Sweden)

    Poulami Basu Thakur

    Full Text Available Pseudomonas syringae pv. syringae B728a, a bacterial pathogen of bean, utilizes large surface populations and extracellular signaling to initiate a fundamental change from an epiphytic to a pathogenic lifestyle. Extracytoplasmic function (ECF sigma (σ factors serve as important regulatory factors in responding to various environmental signals. Bioinformatic analysis of the B728a genome revealed 10 ECF sigma factors. This study analyzed deletion mutants of five previously uncharacterized ECF sigma factor genes in B728a, including three FecI-type ECF sigma factors (ECF5, ECF6, and ECF7 and two ECF sigma factors placed in groups ECF11 and ECF18. Transcriptional profiling by qRT-PCR analysis of ECF sigma factor mutants was used to measure expression of their associated anti-sigma and outer membrane receptor proteins, and expression of genes associated with production of extracellular polysaccharides, fimbriae, glycine betaine and syringomycin. Notably, the B728aΔecf7 mutant displayed reduced swarming and had decreased expression of CupC fimbrial genes. Growth and pathogenicity assays, using a susceptible bean host, revealed that none of the tested sigma factor genes are required for in planta growth and lesion formation.

  9. Identification and functional characterization of Rca1, a transcription factor involved in both antifungal susceptibility and host response in Candida albicans.

    Science.gov (United States)

    Vandeputte, Patrick; Pradervand, Sylvain; Ischer, Françoise; Coste, Alix T; Ferrari, Sélène; Harshman, Keith; Sanglard, Dominique

    2012-07-01

    The identification of novel transcription factors associated with antifungal response may allow the discovery of fungus-specific targets for new therapeutic strategies. A collection of 241 Candida albicans transcriptional regulator mutants was screened for altered susceptibility to fluconazole, caspofungin, amphotericin B, and 5-fluorocytosine. Thirteen of these mutants not yet identified in terms of their role in antifungal response were further investigated, and the function of one of them, a mutant of orf19.6102 (RCA1), was characterized by transcriptome analysis. Strand-specific RNA sequencing and phenotypic tests assigned Rca1 as the regulator of hyphal formation through the cyclic AMP/protein kinase A (cAMP/PKA) signaling pathway and the transcription factor Efg1, but also probably through its interaction with a transcriptional repressor, most likely Tup1. The mechanisms responsible for the high level of resistance to caspofungin and fluconazole observed resulting from RCA1 deletion were investigated. From our observations, we propose that caspofungin resistance was the consequence of the deregulation of cell wall gene expression and that fluconazole resistance was linked to the modulation of the cAMP/PKA signaling pathway activity. In conclusion, our large-scale screening of a C. albicans transcription factor mutant collection allowed the identification of new effectors of the response to antifungals. The functional characterization of Rca1 assigned this transcription factor and its downstream targets as promising candidates for the development of new therapeutic strategies, as Rca1 influences host sensing, hyphal development, and antifungal response.

  10. Characterization of a new CAMP factor carried by an integrative and conjugative element in Streptococcus agalactiae and spreading in Streptococci.

    Directory of Open Access Journals (Sweden)

    Sarah Chuzeville

    Full Text Available Genetic exchanges between Streptococci occur frequently and contribute to their genome diversification. Most of sequenced streptococcal genomes carry multiple mobile genetic elements including Integrative and Conjugative Elements (ICEs that play a major role in these horizontal gene transfers. In addition to genes involved in their mobility and regulation, ICEs also carry genes that can confer selective advantages to bacteria. Numerous elements have been described in S. agalactiae especially those integrated at the 3' end of a tRNA(Lys encoding gene. In strain 515 of S. agalactiae, an invasive neonate human pathogen, the ICE (called 515_tRNA(Lys is functional and carries different putative virulence genes including one encoding a putative new CAMP factor in addition to the one previously described. This work demonstrated the functionality of this CAMP factor (CAMP factor II in Lactococcus lactis but also in pathogenic strains of veterinary origin. The search for co-hemolytic factors in a collection of field strains revealed their presence in S. uberis, S. dysgalactiae, but also for the first time in S. equisimilis and S. bovis. Sequencing of these genes revealed the prevalence of a species-specific factor in S. uberis strains (Uberis factor and the presence of a CAMP factor II encoding gene in S. bovis and S. equisimilis. Furthermore, most of the CAMP factor II positive strains also carried an element integrated in the tRNA(Lys gene. This work thus describes a CAMP factor that is carried by a mobile genetic element and has spread to different streptococcal species.

  11. Characterization of the growth of murine fibroblasts that express human insulin receptors. II. Interaction of insulin with other growth factors

    International Nuclear Information System (INIS)

    Randazzo, P.A.; Jarett, L.

    1990-01-01

    The effects of insulin-like growth factor-1 (IGF-1), epidermal growth factor (EGF), platelet-derived growth factor (PDGF), and insulin on DNA synthesis were studied in murine fibroblasts transfected with an expression vector containing human insulin receptor cDNA (NIH 3T3/HIR) and the parental NIH 3T3 cells. In NIH 3T3/HIR cells, individual growth factors in serum-free medium stimulated DNA synthesis with the following relative efficacies: insulin greater than or equal to 10% fetal calf serum greater than PDGF greater than IGF-1 much greater than EGF. In comparison, the relative efficacies of these factors in stimulating DNA synthesis by NIH 3T3 cells were 10% fetal calf serum greater than PDGF greater than EGF much greater than IGF-1 = insulin. In NIH 3T3/HIR cells, EGF was synergistic with 1-10 ng/ml insulin but not with 100 ng/ml insulin or more. Synergy of PDGF or IGF-1 with insulin was not detected. In the parental NIH 3T3 cells, insulin and IGF-1 were found to be synergistic with EGF (1 ng/ml), PDGF (100 ng/ml), and PDGF plus EGF. In NIH 3T3/HIR cells, the lack of interaction of insulin with other growth factors was also observed when the percentage of cells synthesizing DNA was examined. Despite insulin's inducing only 60% of NIH 3T3/HIR cells to incorporate thymidine, addition of PDGF, EGF, or PDGF plus EGF had no further effect. In contrast, combinations of growth factors resulted in 95% of the parental NIH 3T3 cells synthesizing DNA. The independence of insulin-stimulated DNA synthesis from other mitogens in the NIH 3T3/HIR cells is atypical for progression factor-stimulated DNA synthesis and is thought to be partly the result of insulin receptor expression in an inappropriate context or quantity

  12. Characterization of solid polymer dispersions of active pharmaceutical ingredients by 19F MAS NMR and factor analysis

    Czech Academy of Sciences Publication Activity Database

    Urbanová, Martina; Brus, Jiří; Šeděnková, Ivana; Policianová, Olivia; Kobera, Libor

    2013-01-01

    Roč. 100, 1 January (2013), s. 59-66 ISSN 1386-1425 R&D Projects: GA ČR GPP106/11/P426; GA MŠk 2B08021 Institutional research plan: CEZ:AV0Z40500505 Institutional support: RVO:61389013 Keywords : solid-state NMR * factor analysis * 19F MAS NMR Subject RIV: FR - Pharmacology ; Medidal Chemistry Impact factor: 2.129, year: 2013

  13. Characterization of the primary interaction between the mating pheromone, alpha-factor, and its receptor in Saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    Raths, S.K.

    1987-01-01

    Alpha-factor is a peptide of thirteen amino acids which is required for mating between the haploid mating types, a and α, in Saccharomyces cerevisiae. An analogue of alpha-factor, DHP 8 DHP 11 Nle 12 tridecapeptide, was catalytically reduced in the presence of 3 H gas for production of a radiolabeled pheromone suitable for use in binding studies. Incorporation of tritium resulted in 3 H-alpha-factor with high specific activity, purity, biological activity and long shelf-life. Binding studies revealed that alpha-factor interacts with its receptor via a simple, reversible process which obeys the law of mass action. Association and dissociation kinetics indicate values of 2.92 x 10 6 M/sup /minus/1/ min -1 for k 1 and between 4 and 7 x 10/sup /minus/2/ min/sup /minus/1/ for k/sub /minus/1/. Saturation binding studies reveal an equilibrium dissociation constant equal to 2.32 x 10/sup /minus/8/ M which approximate the kinetically-derived K/sub D/ of 2.12 x 10/sup /minus/8/ M. Scatchard and Hill analyses as well as dissociation behavior in the presence of excess unlabeled ligand indicate alpha-factor interacts with a homogeneous population of binding sites which do not interact and exhibit one affinity for the alpha-factor pheromone

  14. Synthesis and Characterization of Nanodiamond-Growth Factor Complexes Towards Applications in Oral Implantation and Regenerative Medicine.

    Science.gov (United States)

    Bang, Julie; Ting, Caleb; Wang, Peter; Kim, Ted; Wang, Kenneth; Kee, Theodore; Miya, Darron; Ho, Dean; Lee, Dong-Keun

    2018-02-19

    Current challenges in the field of regenerative medicine include the need to deliver sustained concentrations of growth factors and genes that are required to induce the repair of deficient tissues. Enhancement of drug delivery and uptake may result in improved growth factor efficacy. Nanodiamonds (NDs) were explored as potential growth factor delivery agents due to the many favorable properties that they possess. For example, ND's biocompatibility has been extensively validated pre-clinically. In addition, they can be scalably produced through impact events such as detonation. They possess notably faceted surfaces with diverse electrostatic properties that allow the rapid formation of growth factor complexes. In this study, a complex based on NDs conjugated to epidermal growth factor (EGF) functionalized with Alexa Fluor 488 (ND-EGF) was developed. ND-EGF was comprehensively evaluated using dynamic light scattering and zeta potential analysis. Furthermore, the NDs were capable of eluting EGF in a sustained fashion. Therefore, ND-EGF may serve as a promising nano-biomaterial for sustained growth factor elution.

  15. Characterization and in vivo regulon determination of an ECF sigma factor and its cognate anti-sigma factor in Nostoc punctiforme.

    Science.gov (United States)

    Bell, Nicole; Lee, Jamie J; Summers, Michael L

    2017-04-01

    Based on primary sequence comparisons and genomic context, Npun_F4153 (SigG)/Npun_F4154 (SapG) of the cyanobacterium Nostoc punctiforme were hypothesized to encode an ECF sigma factor/anti-sigma factor pair. Transcription of sigG increased in heterocysts and akinetes, and after EDTA treatment. Interaction between SigG and the predicted cytoplasmic domain of SapG was observed in vitro. A SigG-GFP translational fusion protein localized to the periphery of vegetative cells in vivo, but lost this association following heat stress. A sigG mutant was unable to survive envelope damage caused by heat or EDTA, but was able to form functional heterocysts. Akinetes in the mutant strain appeared normal, but these cultures were less resistant to lysozyme and cold treatments than those of the wild-type strain. The SigG in vivo regulon was determined before and during akinete differentiation using DNA microarray analysis, and found to include multiple genes with putative association to the cell envelope. Mapped promoters common to both arrays enabled identification of a SigG promoter-binding motif that was supported in vivo by reporter studies, and in vitro by run-off transcription experiments. These findings support SigG/SapG as a sigma/anti-sigma pair involved in repair of envelope damage resulting from exogenous sources or cellular differentiation. © 2017 John Wiley & Sons Ltd.

  16. Purification and characterization of an inhibitor (soluble tumor necrosis factor receptor) for tumor necrosis factor and lymphotoxin obtained from the serum ultrafiltrates of human cancer patients

    International Nuclear Information System (INIS)

    Gatanaga, Tetsuya; Whang, Chenduen; Cappuccini, F.; Lucci, J.A. III; Jeffes, E.W.B.; Kohr, W.; Lentz, R.; Tomich, J.; Yamamoto, R.S.; Granger, G.A.

    1990-01-01

    Serum ultrafiltrates (SUF) from human patients with different types of cancer contain a blocking factor (BF) that inhibits the cytolytic activity of human tumor necrosis factor α (TNF-α) in vitro. BF is a protein with a molecular mass of 28kDa on reducing sodium dodecyl sulfate/polyacrylamide gel electrophoresis (SDS/PAGE). The active material was purified to homogeneity by a combination of affinity chromatography, PAGE, and high-pressure liquid chromatography. Amino acid sequence analysis revealed that BF is derived from the membrane TNF receptor. Purified BF blocks the lytic activity of recombinant human and mouse TNF-α and recombinant human lymphotoxin activity of TNF-α and recombinant human lymphotoxin on murine L929 cells in vitro. However, BF inhibits the lytic activity of TNF-α more effectively than it does that of lymphotoxin. The BF also inhibits the necrotizing activity of recombinant human TNF-α when coinjected into established cutaneous Meth A tumors in BALB/c mice. The BF may have an important role in (i) the regulation and control of TNF-α and lymphotoxin activity in cancer patients, (ii) interaction between the tumor and the host antitumor mechanisms, and (iii) use of systemically administered TNF-α in clinical trials with human cancer patients

  17. Establishment and characterization of a new cell line, FPS-1, derived from human undifferentiated pleomorphic sarcoma, overexpressing epidermal growth factor receptor and cyclooxygenase-2.

    Science.gov (United States)

    Hakozaki, Michiyuki; Hojo, Hiroshi; Sato, Michiko; Tajino, Takahiro; Yamada, Hitoshi; Kikuchi, Shinichi; Abe, Masafumi

    2006-01-01

    Undifferentiated pleomorphic sarcoma (UPS) is among the most common soft tissue sarcomas in adults. In order to improve its aggressive course or prognosis and establish new therapeutic methods, molecular genetic and biological characterizations of UPS are required. A new human UPS cell line (FPS-1) was established from UPS of the upper arm of a 79-year-old man. The cell line has been maintained for over 14 months with more than 60 passages. FPS-1 cells were characterized using molecular biological methods. FPS-1 cells showed the same morphological and immunophenotypical characteristics as the primary tumor. Cytogenetic and molecular analyses revealed a nonsense mutation in exon 6 of the p53 gene. Epidermal growth factor receptor (EGFR) and cyclooxygenase-2 (COX-2) were expressed in FPS-1 cells. FPS-1 cells might be useful for investigating biological behavior and developing new molecular targeting antitumor drugs for UPS with EGFR or COX-2 expression.

  18. ACCRETA COMPLICATING COMPLETE PLACENTA PREVIA IS CHARACTERIZED BY REDUCED SYSTEMIC LEVELS OF VASCULAR ENDOTHELIAL GROWTH FACTOR AND EPITHELIAL-TO-MESENCHYMAL TRANSITION OF THE INVASIVE TROPHOBLAST

    Science.gov (United States)

    Wehrum, Mark J.; Buhimschi, Irina A.; Salafia, Carolyn; Thung, Stephen; Bahtiyar, Mert O.; Werner, Erica F.; Campbell, Katherine H.; Laky, Christine; Sfakianaki, Anna K.; Zhao, Guomao; Funai, Edmund F.; Buhimschi, Catalin S.

    2011-01-01

    OBJECTIVE To characterize serum angiogenic factor profile of women with complete placenta previa and determine if invasive trophoblast differentiation characteristic of accreta, increta or percreta shares features of epitehelial-mesenchymal-transition (EMT). STUDY DESIGN We analyzed gestational age matched serum samples from 90 pregnant women with either complete placenta previa (n=45) or uncomplicated pregnancies (n=45). Vascular-endothelial-growth-factor (VEGF), placental-growth-factor (PlGF) and soluble fms-like-tyrosine-kinase-1 (sFlt-1) were immunoassayed. VEGF and phosphotyrosine (P-Tyr) immunoreactivity was surveyed in histological specimens relative to expression of vimentin and cytokeratin-7. RESULTS Women with previa and invasive placentation [accreta (n=5); increta (n=6); percreta (n=2)] had lower systemic VEGF (invasive previa: median [IQR]: 0.8[0.02–3.4] vs. control: 6.5[2.7–10.5] pg/mL, P=0.02). VEGF and P-Tyr immunostaining predominated in the invasive extravillous trophoblasts (EVT) which co-expressed vimentin and cytokeratin-7, a EMT feature and tumor-like cell phenotype. CONCLUSIONS Lower systemic free VEGF and a switch of the interstitial EVT to a metastable cell phenotype characterize placenta previa with excessive myometrial invasion. PMID:21316642

  19. Characterization of decontamination factors for evaporators used in the treatment of low and intermediate level liquid radioactive wastes

    International Nuclear Information System (INIS)

    Rood, L.B.; Law, C.G. Jr.

    1972-01-01

    Evaporator decontamination factors were studied as functions of boiloff rate, volume reduction, and feed pH. A bench-scale vertical tube evaporator operating on simulated intermediate level nuclear wastes was used. Decontamination factors were not found to be strong functions of volume reduction or boiloff below vapor velocities of 25 lb/ft 2 -hr. At higher vapor fluxes, splashing was encountered. Foaming occurred at a feed pH of 6 but not at higher values. The presence of radioisotopes in the feed had no effect on evaporator performance

  20. Physico-chemical and mechanical characterization of in-situ forming xyloglucan gels incorporating a growth factor to promote cartilage reconstruction

    International Nuclear Information System (INIS)

    Dispenza, Clelia; Todaro, Simona; Bulone, Donatella; Sabatino, Maria Antonietta; Ghersi, Giulio; San Biagio, Pier Luigi; Lo Presti, Caterina

    2017-01-01

    The development of growth factors is very promising in the field of tissue regeneration but specifically designed formulations have to be developed in order to enable such new biological entities (NBEs). In particular, the range of therapeutic concentrations is usually very low compared to other active proteins and the confinement in the target site can be of crucial importance. In-situ forming scaffolds are very promising solutions for minimally invasive intervention in cartilage reconstruction and targeting of NBEs. In this work injectable, in-situ forming gels of a temperature responsive partially degalactosylated xyloglucan (Deg-XG) incorporating the growth factor FGF-18 are formulated and characterized. In particular, injectability and shear viscosity at room temperature, time-to-gel at body temperature, morphology and mechanical properties of gels are investigated. The highly hydrophobic growth factor is favorably incorporated and retained by the gel. Gels undergo a slow erosion process when immersed in PBS at 37 °C that opens up their porous structure. The prolonged hydrothermal treatment leads to structural rearrangements towards tougher networks with increased dynamic shear modulus. Preliminary biological evaluations confirm absence of cytotoxicity and the ability of these scaffolds to host cells and promote their proliferation. - Highlights: • In-situ forming gels incorporating a growth factor are formulated and characterized. • The gel retains the growth factor and is colonized by chondrocytes. • Mechanical properties and porosity of gels are controlled by polymer concentration. • Incubation at 37 °C increases the gel strength and opens up the porous structure.

  1. Characterization of a Synechocystis double mutant lacking the photosystem II assembly factors YCF48 and Sll0933

    Czech Academy of Sciences Publication Activity Database

    Rengstl, B.; Knoppová, Jana; Komenda, Josef; Nickelsen, J.

    2013-01-01

    Roč. 237, č. 2 (2013), s. 471-480 ISSN 0032-0935 R&D Projects: GA ČR GBP501/12/G055; GA MŠk(CZ) ED2.1.00/03.0110 Institutional support: RVO:61388971 Keywords : Complex assembly * Cyanobacteria * Photosynthesis Subject RIV: EE - Microbiology, Virology Impact factor: 3.376, year: 2013

  2. Fluxing as a new tool for bitumen rheological characterization and the use of time-concentration shift factor (ac)

    CSIR Research Space (South Africa)

    Zoorob, SE

    2018-01-01

    Full Text Available he concept of temperature shift factor (aT) as defined by Doolittle, relating the free volume of a viscoelastic material at the current and reference states is briefly examined together with the resultant William-Landel-Ferry equation. This paper...

  3. Molecular cloning and functional characterization of the human platelet-derived growth factor alpha receptor gene promoter

    NARCIS (Netherlands)

    Afink, G. B.; Nistér, M.; Stassen, B. H.; Joosten, P. H.; Rademakers, P. J.; Bongcam-Rudloff, E.; van Zoelen, E. J.; Mosselman, S.

    1995-01-01

    Expression of the platelet-derived growth factor alpha receptor (PDGF alpha R) is strictly regulated during mammalian development and tumorigenesis. The molecular mechanisms involved in the specific regulation of PDGF alpha R expression are unknown, but transcriptional regulation of the PDGF alpha R

  4. Characterization of the domains of E. coli initiation factor IF2 responsible for recognition of the ribosome

    DEFF Research Database (Denmark)

    Manuel Palacios Moreno, Juan; Andersen, Lars Dyrskjøt; Egebjerg Kristensen, Janni

    1999-01-01

    We have studied the interactions between the ribosome and the domains of Escherichia coli translation initiation factor 2, using an in vitro ribosomal binding assay with wild-type forms, N- and C-terminal truncated forms of IF2 as well as isolated structural domains. A deletion mutant of the fact...

  5. 3-Way characterization of soils by Procrustes rotation, matrix-augmented principal components analysis and parallel factor analysis

    Czech Academy of Sciences Publication Activity Database

    Andrade, J.M.; Kubista, Mikael; Carlosena, A.; Prada, D.

    2007-01-01

    Roč. 603, č. 1 (2007), s. 20-29 ISSN 0003-2670 Institutional research plan: CEZ:AV0Z50520514 Keywords : PCA * heavy metals * soil Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.186, year: 2007

  6. Cloning and characterization of p52, the fifth subunit of the core of transcription/repair factor TFIIH.

    NARCIS (Netherlands)

    J.C. Marinoni; R. Roy (Richard); W. Vermeulen (Wim); P. Miniou; Y. Lutz; G. Weeda (Geert); T. Seroz; D.M. Gomez (Denise Molina); J.H.J. Hoeijmakers (Jan); J-M. Egly (Jean-Marc)

    1997-01-01

    textabstractTFIIH is a multiprotein factor involved in transcription and DNA repair and is implicated in DNA repair/transcription deficiency disorders such as xeroderma pigmentosum, Cockayne syndrome and trichothiodystrophy. Eight out of the nine genes encoding the subunits forming TFIIH have

  7. Characterization of cutaneous vascular permeability induced by platelet-activating factor in guinea pigs and rats and its inhibition by a platelet-activating factor receptor antagonist

    International Nuclear Information System (INIS)

    Hwang, S.B.; Li, C.L.; Lam, M.H.; Shen, T.Y.

    1985-01-01

    Mechanisms of platelet-activating factor (PAF)-induced increases of cutaneous vascular permeability in guinea pigs and in rats were further explored. PAF so far is the most potent vasoactive mediator, being more than 1000-fold more potent than histamine and bradykinin in both species. In guinea pigs, there is a time delay of 5 to 10 minutes before PAF action, whereas, in the rat, the increased vasopermeability occurs immediately following the intradermal PAF injection. Relative vasoactive potencies of PAF and several structure-related analogues in both species correlate very well with their relative inhibition of the binding of 3 H-PAF to specific receptor sites on isolated rabbit platelet plasma membranes and their aggregatory abilities of rabbit platelets. Furthermore, the PAF-induced cutaneous vascular permeability is inhibitable by a competitive specific PAF receptor antagonist, kadsurenone, suggesting that binding of PAF to its specific receptor site is the first step to initiate its action of increased cutaneous vascular permeability. Several pure cyclooxygenase inhibitors, including indomethacin, diflunisal, and flurbiprofen, and the dual cyclooxygenase/lipoxygenase inhibitor, BW755C, but not the histamine antagonists, inhibit the PAF-induced vasopermeability in guinea pigs. The inhibition by indomethacin or BW755C can be fully reversed by coinjection intradermally with PAF and prostaglandin E1 but not leukotriene B4. Also, prostaglandin E1 but not leukotriene B4 enhances the guinea pig in vivo response to PAF in this model. However, in rats, none of the cyclooxygenase inhibitors, histamine antagonists, or BW755C inhibit the PAF effect of cutaneous phenomena

  8. Genomic characterization, phylogenetic analysis, and identification of virulence factors in Aerococcus sanguinicola and Aerococcus urinae strains isolated from infection episodes

    DEFF Research Database (Denmark)

    Carkaci, Derya; Højholt, Katrine; Nielsen, Xiaohui Chen

    2017-01-01

    Aerococcus sanguinicola and Aerococcus urinae are emerging pathogens in clinical settings mostly being causative agents of urinary tract infections (UTIs), urogenic sepsis and more seldomly complicated infective endocarditis (IE). Limited knowledge exists concerning the pathogenicity of these two...... species. Eight clinical A. sanguinicola (isolated from 2009 to 2015) and 40 clinical A. urinae (isolated from 1984 to 2015) strains from episodes of UTIs, bacteremia, and IE were whole-genome sequenced (WGS) to analyze genomic diversity and characterization of virulence genes involved in the bacterial....... In conclusion, this is the first study dealing with WGS and comparative genomics of clinical A. sanguinicola and A. urinae strains from episodes of UTIs, bacteremia, and IE. Gene homologs associated with antiphagocytosis and bacterial adherence were identified and genetic variability was observed within A...

  9. Characterization of shark complement factor I gene(s): genomic analysis of a novel shark-specific sequence.

    Science.gov (United States)

    Shin, Dong-Ho; Webb, Barbara M; Nakao, Miki; Smith, Sylvia L

    2009-07-01

    Complement factor I is a crucial regulator of mammalian complement activity. Very little is known of complement regulators in non-mammalian species. We isolated and sequenced four highly similar complement factor I cDNAs from the liver of the nurse shark (Ginglymostoma cirratum), designated as GcIf-1, GcIf-2, GcIf-3 and GcIf-4 (previously referred to as nsFI-a, -b, -c and -d) which encode 689, 673, 673 and 657 amino acid residues, respectively. They share 95% (shark-specific sequence between the leader peptide (LP) and the factor I membrane attack complex (FIMAC) domain. The cDNA sequences differ only in the size and composition of the shark-specific region (SSR). Sequence analysis of each SSR has identified within the region two novel short sequences (SS1 and SS2) and three repeat sequences (RS1-3). Genomic analysis has revealed the existence of three introns between the leader peptide and the FIMAC domain, tentatively designated intron 1, intron 2, and intron 3 which span 4067, 2293 and 2082bp, respectively. Southern blot analysis suggests the presence of a single gene copy for each cDNA type. Phylogenetic analysis suggests that complement factor I of cartilaginous fish diverged prior to the emergence of mammals. All four GcIf cDNA species are expressed in four different tissues and the liver is the main tissue in which expression level of all four is high. This suggests that the expression of GcIf isotypes is tissue-dependent.

  10. Cloning and characterization of cDNAs encoding the complete sequence of decay-accelerating factor of human complement

    International Nuclear Information System (INIS)

    Medof, M.E.; Lublin, D.M.; Holers, V.M.; Ayers, D.J.; Getty, R.R.; Leykam, J.F.; Atkinson, J.P.; Tykocinski, M.L.

    1987-01-01

    cDNAs encoding the complement decay-accelerating factor (DAF) were isolated from HeLa and differentiated HL-60 λgt cDNA libraries by screening with a codon preference oligonucleotide corresponding to DAF NH 2 -terminal amino acids 3-14. The composite cDNA sequence showed a 347-amino acid protein preceded by an NH 2 -terminal leader peptide sequence. The translated sequence beginning at the DAF NH 2 terminus encodes four contiguous ≅ 61-amino acid long repetitive units of internal homology. The repetitive regions contain four conserved cysteines, one proline, one glycine, one glycine/alanine, four leucines/isoleucines/valines, one serine, three tyrosines/phenylalanines, and on tryptophan and show striking homology to similar regions previously identified in factor B, C2, C4 binding protein, factor H, C1r, factor XIII, interleukin 2 receptor, and serum β 2 -glycoprotein I. The consensus repeats are attached to a 70-amino acid long segment rich in serine and threonine (potential O-glycosylation sites), which is in turn followed by a stretch of hydrophobic amino acids. RNA blot analysis of HeLa and HL-60 RNA revealed three DAF mRNA species of 3.1, 2.7, and 2.0 kilobases. The results indicate that portions of the DAF gene may have evolved from a DNA element common to the above proteins, that DAF cDNA predicts a COOH-terminal anchoring polypeptide, and that distinct species of DAF message are elaborated in cells

  11. Characterization of air pollutant concentrations, fleet emission factors, and dispersion near a North Carolina interstate freeway across two seasons

    Science.gov (United States)

    Saha, Provat K.; Khlystov, Andrey; Snyder, Michelle G.; Grieshop, Andrew P.

    2018-03-01

    We present field measurement data and modeling of multiple traffic-related air pollutants during two seasons at a site adjoining Interstate 40, near Durham, North Carolina. We analyze spatial-temporal and seasonal trends and fleet-average pollutant emission factors and use our data to evaluate a line source dispersion model. Month-long measurement campaigns were performed in summer 2015 and winter 2016. Data were collected at a fixed near-road site located within 10 m from the highway edge, an upwind background site and, under favorable meteorological conditions, along downwind perpendicular transects. Measurements included the size distribution, chemical composition, and volatility of submicron particles, black carbon (BC), nitrogen oxides (NOx), meteorological conditions and traffic activity data. Results show strong seasonal and diurnal differences in spatial distribution of traffic sourced pollutants. A strong signature of vehicle emissions was observed within 100-150 m from the highway edge with significantly higher concentrations during morning. Substantially higher concentrations and less-sharp near-road gradients were observed in winter for many species. Season-specific fleet-average fuel-based emission factors for NO, NOx, BC, and particle number (PN) were derived based on up- and down-wind roadside measurements. The campaign-average NOx and PN emission factors were 20% and 300% higher in winter than summer, respectively. These results suggest that the combined effect of higher emissions and their slower downwind dispersion in winter dictate the observed higher downwind concentrations and wider highway influence zone in winter for several species. Finally, measurements of traffic data, emission factors, and pollutant concentrations were integrated to evaluate a line source dispersion model (R-LINE). The dispersion model captured the general trends in the spatial and temporal patterns in near-road concentrations. However, there was a tendency for the model

  12. A characterization of factors determining postoperative ileus after laparoscopic colectomy enables the generation of a novel predictive score.

    Science.gov (United States)

    Kronberg, Udo; Kiran, Ravi P; Soliman, Mohamed S M; Hammel, Jeff P; Galway, Ursula; Coffey, John Calvin; Fazio, Victor W

    2011-01-01

    Postoperative ileus (POI) after colorectal surgery is associated with prolonged hospital stay and increased costs. The aim of this study is to investigate pre-, intra-, and postoperative risk factors associated with the development of POI in patients undergoing laparoscopic partial colectomy. Patients operated between 2004 and 2008 were retrospectively identified from a prospectively maintained database, and clinical, metabolic, and pharmacologic data were obtained. Postoperative ileus was defined as the absence of bowel function for 5 or more days or the need for reinsertion of a nasogastric tube after starting oral diet in the absence of mechanical obstruction. Associations between likelihood of POI and study variables were assessed univariably by using χ tests, Fisher exact tests, and logistic regression models. A scoring system for prediction of POI was constructed by using a multivariable logistic regression model based on forward stepwise selection of preoperative factors. A total of 413 patients (mean age, 58 years; 53.5% women) were included, and 42 (10.2%) of them developed POI. Preoperative albumin, postoperative deep-vein thrombosis, and electrolyte levels were associated with POI. Age, previous abdominal surgery, and chronic preoperative use of narcotics were independently correlated with POI on multivariate analysis, which allowed the creation of a predictive score. Patients with a score of 2 or higher had an 18.3% risk of POI (P POI can be predicted by using a preoperative scoring system. Addressing the postoperative factors may be expected to reduce the incidence of this common complication in high-risk patients.

  13. Epidemiological characterization and risk factors associated with Mycobacterium avium subsp. paratuberculosis infection in dairy goats in the Brazilian semiarid region

    Directory of Open Access Journals (Sweden)

    Theonys Diógenes Freitas

    2015-02-01

    Full Text Available The aim of this investigation was to conduct an epidemiological study and identify risk factors associated with the occurrence of paratuberculosis (Johne’s disease in dairy goats within the semiarid region of Paraíba State. The study was done during the period of March 2009 to July 2011, during which 727 female goats from 86 flocks from the city of Monteiro, Paraíba were investigated. For the serological diagnosis of Mycobacterium avium subsp. paratuberculosis (Map infection indirect ELISA tests (screening and confirmatory were performed. Of the 727 animals used six (0.82% were seropositive at the confirmatory test after screening, and of the 86 flocks six (6.97% presented at least one seropositive animal. In positive flocks the frequency of reactive animals ranged from 5.26% to 16.60%. Risk factors identified were production system (weaning and reproduction (odds ratio = 36.0; 95% CI = 2.6 –486.1; p < 0,001 and absence of technical infrastructure (odds ratio = 54.0; 95% CI = 4.5 –642.9; p < 0,001. It was concluded that Mycobacterium avium subsp. paratuberculosis is present in dairy goat flocks in the region; however, its influence on decrease productivity as well as the risk of transmission to humans through animal products must totally evaluated. Based on the analysis of risk factors, improvements are recommended for the technical infrastructure and the management of breeding goats.

  14. Human umbilical cord blood-derived stem cells and brain-derived neurotrophic factor protect injured optic nerve: viscoelasticity characterization

    Directory of Open Access Journals (Sweden)

    Xue-man Lv

    2016-01-01

    Full Text Available The optic nerve is a viscoelastic solid-like biomaterial. Its normal stress relaxation and creep properties enable the nerve to resist constant strain and protect it from injury. We hypothesized that stress relaxation and creep properties of the optic nerve change after injury. More-over, human brain-derived neurotrophic factor or umbilical cord blood-derived stem cells may restore these changes to normal. To validate this hypothesis, a rabbit model of optic nerve injury was established using a clamp approach. At 7 days after injury, the vitreous body re-ceived a one-time injection of 50 µg human brain-derived neurotrophic factor or 1 × 106 human umbilical cord blood-derived stem cells. At 30 days after injury, stress relaxation and creep properties of the optic nerve that received treatment had recovered greatly, with patho-logical changes in the injured optic nerve also noticeably improved. These results suggest that human brain-derived neurotrophic factor or umbilical cord blood-derived stem cell intervention promotes viscoelasticity recovery of injured optic nerves, and thereby contributes to nerve recovery.

  15. High level of Brazilian men´s volleyball: characterization and difference of predictive factors of back row attack

    Directory of Open Access Journals (Sweden)

    Gustavo de Conti Teixeira Costa Conti

    2018-05-01

    Full Text Available This study aimed to identify the predictive factors of attacks, performed from positions 1 and 6 according to the effect of reception in high level Brazilian male volleyball and to find the predictive factors that differentiate the game practiced from these positions. The sample consisted in the observation of 142 games of the Brazilian Men's Super League, totalling 2969 actions of reception, setting and attack from positions 1 and 6. The significance value adopted was 5% (p ≤ 0.05. The analysis of the predictive factors of the game performed by the attacker of position 1 showed greater chances to score after an excellent (odds ratio adjusted – ORA = 1.48 and moderate effect of reception (ORA = 1.31, the second attack tempo (ORA = 1.32, the powerful attack in parallel (ORA = 1.91 and in diagonal (ORA =3.44. The attacker of position 6 showed higher chances of scoring after a high effect of reception (ORA = 3.39 and powerful attack in the parallel (ORA = 1.53. In conclusion, regardless the effect of reception, the use of the back-row attackers is recommended to increase the uncertainty on the opposing team and the chances to score.

  16. Contamination of public transports by Staphylococcus aureus and its carriage by biomedical students: point-prevalence, related risk factors and molecular characterization of methicillin-resistant strains.

    Science.gov (United States)

    Mendes, Â; Martins da Costa, P; Rego, D; Beça, N; Alves, C; Moreira, T; Conceição, T; Aires-de-Sousa, M

    2015-08-01

    To analyse the contamination of public transports by Staphylococcus aureus and assess its carriage by biomedical students, focussing on the point-prevalence, related risk factors and molecular characterization of methicillin-resistant strains. Cross-sectional survey. Methicillin-resistant S. aureus (MRSA) and methicillin-sensitive S. aureus (MSSA) isolated from handrails of buses (n = 112) and trains (n = 79) circulating in Porto and from nasal swabs of local university students (n = 475) were quantified, characterized by molecular typing methods and related to possible risk factors. The MRSA prevalence in buses (16.1%) was not significantly different from trains (8.9%). There was also no identifiable association between the counts of MSSA and MRSA in buses and trains and the number of travellers in each sampling day, specific routes (including those passing by main hospitals) or other risk factors. Of the students, 37.1% carried S. aureus, and having a part-time job or smoking were found to be risk factors for carriage. EMRSA-15 (ST22-SCCmecIVh) was the prevalent MRSA clonal lineage, found not only in the buses (n = 14) and trains (n = 2) but also in the single MRSA-carrier among the students. The characteristics of the community-associated Southwest Pacific MRSA clone were found in a single ST30-IVa isolate, which may suggest a recent SCCmec acquisition by an MSSA background in the community. The spread of EMRSA-15, a common hospital-associated lineage, among different public transports and as a nasal coloniser is of concern and warrants adequate public health control measures. Copyright © 2015 The Royal Society for Public Health. Published by Elsevier Ltd. All rights reserved.

  17. A multi-factor approach for process-based seabed characterization: example from the northeastern continental margin of the Korean peninsula (East Sea)

    Science.gov (United States)

    Cukur, Deniz; Um, In-Kwon; Chun, Jong-Hwa; Kim, So-Ra; Lee, Gwang-Soo; Kim, Yuri; Kong, Gee-Soo; Horozal, Senay; Kim, Seong-Pil

    2018-04-01

    This study investigates sediment transport and depositional processes from a newly collected dataset comprising sub-bottom chirp profiles, multibeam bathymetry, and sediment cores from the northeastern continental margin of Korea in the East Sea (Japan Sea). Twelve echo-types and eleven sedimentary facies have been defined and interpreted as deposits formed by shallow-marine, hemipelagic sedimentation, bottom current, and mass-movement processes. Hemipelagic sedimentation, which is acoustically characterized by undisturbed layered sediments, appears to have been the primary sedimentary process throughout the study area. The inner and outer continental shelf (shallow-marine sedimentary processes. Two slope-parallel canyons, 0.2-2 km wide and up to 30 km long, appear to have acted as possible conduits for turbidity currents from the shallower shelf into the deep basins. Bottom current deposits, expressed as erosional moats immediately below topographic highs, are prevalent on the southern lower slope at water depths of 400-450 m. Mass-movements (i.e., slides/slumps, debris flow deposits) consisting of chaotic facies characterize the lower slope and represent one of the most important sedimentary processes in the study area. Piston cores confirm the presence of mass-transport deposits (MTDs) that are characterized by mud clasts of variable size, shape, and color. Multibeam bathymetry shows that large-scale MTDs are chiefly initiated on the lower slope (400-600 m) with gradients up to 3° and where they produce scarps on the order of 100 m in height. Sandy MTDs also occur on the upper continental slope adjacent to the seaward edge of the shelf terrace. Earthquakes associated with tectonic activity and the development of fluid overpressure is considered as the main conditioning factor for destabilizing the slope sediments. Overall, the sedimentary processes show typical characteristics of a fine-grained clastic slope apron and change down-slope and differ within each

  18. Characterization of a novel mutation in the von Willebrand factor propeptide in a distinct subtype of recessive von Willebrand disease

    DEFF Research Database (Denmark)

    Lanke, Elsa; Kristoffersson, Ann-Charlotte; Philips, Malou

    2008-01-01

    von Willebrand factor (VWF) is a plasma protein that consists of a series of multimers of which the high-molecular-weight VWF multimers are the most potent in platelet adhesion and aggregation. The propeptide of the VWF (VWFpp) is known to be essential in the process of multimer assembly. Genetic...... mutation in the VWFpp abolishes multimerization of VWF. The mutation probably disrupts the normal configuration of the VWFpp, which is essential for correct orientation of the protomers and ultimately multimerization. The mutant amino acid is located in a region that is highly conserved across several...

  19. Principle of an operational complexity index for the characterization of the human factor relevance of future reactors concepts

    International Nuclear Information System (INIS)

    Papin, Bernard

    2004-01-01

    With the increasing reliability of the modern technological systems, the human contribution to the global risk in the operation of industrial systems is becoming more and more significant : in the nuclear reactor operation for example, a recent PSA estimation of this contribution is about 25% of the risk of core melting, all situations considered. This urges the designers of future nuclear reactors to consider the minimisation of this Human Factor (HF) contribution, at the very early stage of their design : the experience feedback shows that this is indeed at this stage that the fundamental design options, impacting the most the human reliability in operation, are fixed. The problem is that at these early design stages, it is also quite impossible to apply formal human reliability methods to support this HF optimisation, while the precise operating conditions of the reactor are not yet known in enough details. In this paper, another approach of the HF evaluation during the design, based on the functional and operational complexity assessment, is proposed. As an illustration, this approach is used to compare various concepts of Pressurized Water Reactors from the point of view of the Human Factor relevance. (Author)

  20. Fabrication and characterization of a novel microparticle with gyrus-patterned surface and growth factor delivery for cartilage tissue engineering

    Energy Technology Data Exchange (ETDEWEB)

    Huang Sha [Department of Oral Histology and Pathology, School of Stomatology, Fourth Military Medical University, Xi' an 710032 (China); Research and Development Center for Tissue Engineering, Fourth Military Medical University, Xi' an 710032 (China); Wang Yijuan [Key Laboratory for Macromolecular Science of Shaanxi Province, Shaanxi Normal University, Xi' an 710062 (China); Liang Tang [Department of Oral Histology and Pathology, School of Stomatology, Fourth Military Medical University, Xi' an 710032 (China); Research and Development Center for Tissue Engineering, Fourth Military Medical University, Xi' an 710032 (China); Jin Fang [Department of Orthodontics, School of Stomatology, Fourth Military Medical University, Xi' an 710032 (China); Liu Shouxin [Key Laboratory for Macromolecular Science of Shaanxi Province, Shaanxi Normal University, Xi' an 710062 (China); Jin Yan, E-mail: yanjin@fmmu.edu.cn [Department of Oral Histology and Pathology, School of Stomatology, Fourth Military Medical University, Xi' an 710032 (China); Research and Development Center for Tissue Engineering, Fourth Military Medical University, Xi' an 710032 (China)

    2009-05-05

    Microparticles can serve as substrates for cell amplification and deliver the expanded cells to the site of the defect. It was hypothesized that a novel microparticle combined of sustained and localized delivery of proliferative growth factors and gyrus-patterned surface would influence the cell behaviours of adherence and expansion on the microparticle in the present study. To test the hypothesis, gelatin particles with diameter ranging from 280 to 350 {mu}m were fabricated and were modified by cryogenic freeze-drying treatment and basic fibroblast growth factor (bFGF) incorporation. The results of in vitro chondrocyte culture illustrated that cells could proliferate more obviously on the microparticles with bFGF addition, but no correlation between attachment rate and bFGF was observed. On the other hand, microparticles with gyrus-patterned surface demonstrated the highest cell attachment rate and higher rate of cell growth, in particular on bFGF combined ones. It seems to be a promising candidate as a chondrocyte microparticle and could be the potential application in cartilage tissue engineering.

  1. Identification and characterization of bZIP-type transcription factors involved in carrot (Daucus carota L.) somatic embryogenesis.

    Science.gov (United States)

    Guan, Yucheng; Ren, Haibo; Xie, He; Ma, Zeyang; Chen, Fan

    2009-10-01

    Seed dormancy is an important adaptive trait that enables seeds of many species to remain quiescent until conditions become favorable for germination. Abscisic acid (ABA) plays an important role in these developmental processes. Like dormancy and germination, the elongation of carrot somatic embryo radicles is retarded by sucrose concentrations at or above 6%, and normal growth resumes at sucrose concentrations below 3%. Using a yeast one-hybrid screening system, we isolated two bZIP-type transcription factors, CAREB1 and CAREB2, from a cDNA library prepared from carrot somatic embryos cultured in a high-sucrose medium. Both CAREB1 and CAREB2 were localized to the nucleus, and specifically bound to the ABA response element (ABRE) in the Dc3 promoter. Expression of CAREB2 was induced in seedlings by drought and exogenous ABA application; whereas expression of CAREB1 increased during late embryogenesis, and reduced dramatically when somatic embryos were treated with fluridone, an inhibitor of ABA synthesis. Overexpression of CAREB1 caused somatic embryos to develop slowly when cultured in low-sucrose medium, and retarded the elongation of the radicles. These results indicate that CAREB1 and CAREB2 have similar DNA-binding activities, but play different roles during carrot development. Our results indicate that CAREB1 functions as an important trans-acting factor in the ABA signal transduction pathway during carrot somatic embryogenesis.

  2. Fabrication and characterization of a novel microparticle with gyrus-patterned surface and growth factor delivery for cartilage tissue engineering

    International Nuclear Information System (INIS)

    Huang Sha; Wang Yijuan; Liang Tang; Jin Fang; Liu Shouxin; Jin Yan

    2009-01-01

    Microparticles can serve as substrates for cell amplification and deliver the expanded cells to the site of the defect. It was hypothesized that a novel microparticle combined of sustained and localized delivery of proliferative growth factors and gyrus-patterned surface would influence the cell behaviours of adherence and expansion on the microparticle in the present study. To test the hypothesis, gelatin particles with diameter ranging from 280 to 350 μm were fabricated and were modified by cryogenic freeze-drying treatment and basic fibroblast growth factor (bFGF) incorporation. The results of in vitro chondrocyte culture illustrated that cells could proliferate more obviously on the microparticles with bFGF addition, but no correlation between attachment rate and bFGF was observed. On the other hand, microparticles with gyrus-patterned surface demonstrated the highest cell attachment rate and higher rate of cell growth, in particular on bFGF combined ones. It seems to be a promising candidate as a chondrocyte microparticle and could be the potential application in cartilage tissue engineering.

  3. Presenting a mapping method based on fuzzy Logic and TOPSIS multi criteria decision-making methods to detect promising porphyry copper mineralization areas in the east of the Sarcheshmeh copper metallogenic district

    Directory of Open Access Journals (Sweden)

    Shokouh Riahi

    2017-11-01

    through applying the multi-criteria decision-making method. Finally, the selected favorable areas in the metallogenic strip located at the south to the south-east of the Sarcheshmeh porphyry copper deposit are prioritized and introduced for further follow up ground exploration operations. Methodology In order to solve complex decision-making problems like the problem of mapping favorable porphyry copper mineralization zones under great uncertainties, the TOPSIS method is considered as an appropriate approach offering significant simplicity, flexibility and capability (Ataei., 2010. The TOPSIS method is considered to be an efficient method due to having very high accuracy, speed, sensitivity as well as being easy to implement and interpret the outputted results (Hwang and Yoon, 1981. It has found many applications in important areas of mining industry where there is a need to make decisions under risky conditions and data uncertainties. One basic issue in applying decision-making methods in the field of mineral exploration is to rank and propose the best possible candidates among all potentially favorable areas for the next stage of mineral exploration. In this regard, the best favorable areas are selected based on exploratory data layers including favorable lithologies, alterations, structures plus geochemical and geophysical anomalies (Pazand et al., 2012. Results and discussion In the first step, the area located south to the southeast of one the largest porphyry copper deposits in Iran known as Sarcheshmeh was investigated for favorable areas using all available exploratory data as mentioned in the previous section using fuzzy logic integration approach in the GIS environment. Evaluating the highly favorable areas presented by the fuzzy logic approach showed great consistency with the already known copper mineralization prospects. Next, the first 20 priorities obtained from the fuzzy logic approach were chosen as the best candidates to be ranked using the TOPSIS multi

  4. Corticotropin-releasing factor (CRF) receptors in intermediate lobe of the pituitary: Biochemical characterization and autoradiographic localization

    International Nuclear Information System (INIS)

    Grigoriadis, D.E.; De Souza, E.B.

    1989-01-01

    CRF receptors were characterized using radioligand binding and chemical affinity cross-linking techniques and localized using autoradiographic techniques in porcine, bovine and rat pituitaries. The binding of 125I-[Tyr0]-ovine CRF (125I-oCRF) to porcine anterior and neurointermediate lobe membranes was saturable and of high affinity with comparable KD values (200-600 pM) and receptor densities (100-200 fmoles/mg protein). The pharmacological rank order of potencies for various analogs and fragments of CRF in inhibiting 125I-oCRF binding in neurointermediate lobe was characteristic of the well-established CRF receptor in anterior pituitary. Furthermore, the binding of 125I-oCRF to both anterior and neurointermediate lobes of the pituitary was guanine nucleotide-sensitive. Affinity cross-linking studies revealed that the molecular weight of the CRF binding protein in rat intermediate lobe was identical to that in rat anterior lobe (Mr = 75,000). While the CRF binding protein in the anterior lobes of porcine and bovine pituitaries had identical molecular weights to CRF receptors in rat pituitary (Mr = 75,000), the molecular weight of the CRF binding protein in porcine and bovine intermediate lobe was slightly higher (Mr = 78,000). Pituitary autoradiograms from the three species showed specific binding sites for 125I-oCRF in anterior and intermediate lobes, with none being apparent in the posterior pituitary. The identification of CRF receptors in the intermediate lobe with comparable characteristics to those previously identified in the anterior pituitary substantiate further the physiological role of CRF in regulating intermediate lobe hormone secretion

  5. Functional characterization of replication and stability factors of an incompatibility group P-1 plasmid from Xylella fastidiosa.

    Science.gov (United States)

    Lee, Min Woo; Rogers, Elizabeth E; Stenger, Drake C

    2010-12-01

    Xylella fastidiosa strain riv11 harbors a 25-kbp plasmid (pXF-RIV11) belonging to the IncP-1 incompatibility group. Replication and stability factors of pXF-RIV11 were identified and used to construct plasmids able to replicate in X. fastidiosa and Escherichia coli. Replication in X. fastidiosa required a 1.4-kbp region from pXF-RIV11 containing a replication initiation gene (trfA) and the adjacent origin of DNA replication (oriV). Constructs containing trfA and oriV from pVEIS01, a related IncP-1 plasmid of the earthworm symbiont Verminephrobacter eiseniae, also were competent for replication in X. fastidiosa. Constructs derived from pXF-RIV11 but not pVEIS01 replicated in Agrobacterium tumefaciens, Xanthomonas campestris, and Pseudomonas syringae. Although plasmids bearing replication elements from pXF-RIV11 or pVEIS01 could be maintained in X. fastidiosa under antibiotic selection, removal of selection resulted in plasmid extinction after 3 weekly passages. Addition of a toxin-antitoxin addiction system (pemI/pemK) from pXF-RIV11 improved plasmid stability such that >80 to 90% of X. fastidiosa cells retained plasmid after 5 weekly passages in the absence of antibiotic selection. Expression of PemK in E. coli was toxic for cell growth, but toxicity was nullified by coexpression of PemI antitoxin. Deletion of N-terminal sequences of PemK containing the conserved motif RGD abolished toxicity. In vitro assays revealed a direct interaction of PemI with PemK, suggesting that antitoxin activity of PemI is mediated by toxin sequestration. IncP-1 plasmid replication and stability factors were added to an E. coli cloning vector to constitute a stable 6.0-kbp shuttle vector (pXF20-PEMIK) suitable for use in X. fastidiosa.

  6. Characterization of virulence factor regulation by SrrAB, a two-component system in Staphylococcus aureus.

    Science.gov (United States)

    Pragman, Alexa A; Yarwood, Jeremy M; Tripp, Timothy J; Schlievert, Patrick M

    2004-04-01

    Workers in our laboratory have previously identified the staphylococcal respiratory response AB (SrrAB), a Staphylococcus aureus two-component system that acts in the global regulation of virulence factors. This system down-regulates production of agr RNAIII, protein A, and toxic shock syndrome toxin 1 (TSST-1), particularly under low-oxygen conditions. In this study we investigated the localization and membrane orientation of SrrA and SrrB, transcription of the srrAB operon, the DNA-binding properties of SrrA, and the effect of SrrAB expression on S. aureus virulence. We found that SrrA is localized to the S. aureus cytoplasm, while SrrB is localized to the membrane and is properly oriented to function as a histidine kinase. srrAB has one transcriptional start site which results in either an srrA transcript or a full-length srrAB transcript; srrB must be cotranscribed with srrA. Gel shift assays of the agr P2, agr P3, protein A (spa), TSST-1 (tst), and srr promoters revealed SrrA binding at each of these promoters. Analysis of SrrAB-overexpressing strains by using the rabbit model of bacterial endocarditis demonstrated that overexpression of SrrAB decreased the virulence of the organisms compared to the virulence of isogenic strains that do not overexpress SrrAB. We concluded that SrrAB is properly localized and oriented to function as a two-component system. Overexpression of SrrAB, which represses agr RNAIII, TSST-1, and protein A in vitro, decreases virulence in the rabbit endocarditis model. Repression of these virulence factors is likely due to a direct interaction between SrrA and the agr, tst, and spa promoters.

  7. [Factor Characterization Associated with Suicidal Behavior in 8(th) Grade Adolescent Students in Three Schools from Bogotá (Colombia)].

    Science.gov (United States)

    Pérez-Olmos, Isabel; Cruz, Doris Lorena Téllez; Traslaviña, Ángela Liliana Vélez; Ibáñez-Pinilla, Milcíades

    2012-03-01

    To establish the probability for suicide risk and/or mental disorders, together with related factors among high school students in 3 schools in Bogota. Cross sectional study of 309 adolescents. The average age was 13.83 ± 0.9, female dominance (58.6%) and a 3(rd) socioeconomic stratum (68.3%). The suicidal risk behavioral probability and/or mental symptoms was 47.6%, 26.5% exhibited some suicide manifestations, 14.23% had experienced suicidal ideas in the last 3 months, 3.55% had had suicide attempts at least once in life, and 8.73% had suicidal ideas in the last 3 months with suicide attempts. The risk of suicidal behavior and/or mental disorders was explained jointly by depression (OR=27.9, 95% CI: 3.5-223. 1), low self-esteem (OR=11.8, 95% CI: 2.5-56.5), severe family dysfunction (OR=3.4, 95%CI 1.2-9.7), being female (OR=2.1, 95% CI: 1.2-3.8) and being 15 or older (OR=1.9, 95% CI: 0.967-3.9). Psychological abuse followed by physical mistreatment was associated with suicidal behavior and /or mental illness while good family relationships were associated to lower probability. Depression, low self-esteem, severe family dysfunction, female gender, older age (≥15) and domestic violence are risk factors associated with suicide and/or mental disorders in adolescents; good family relationships are associated with lower risk. Copyright © 2012 Asociación Colombiana de Psiquiatría. Publicado por Elsevier España. All rights reserved.

  8. Genome-Wide Identification and Characterization of BrrTCP Transcription Factors in Brassica rapa ssp. rapa

    Directory of Open Access Journals (Sweden)

    Jiancan Du

    2017-09-01

    Full Text Available The teosinte branched1/cycloidea/proliferating cell factor (TCP gene family is a plant-specific transcription factor that participates in the control of plant development by regulating cell proliferation. However, no report is currently available about this gene family in turnips (Brassica rapa ssp. rapa. In this study, a genome-wide analysis of TCP genes was performed in turnips. Thirty-nine TCP genes in turnip genome were identified and distributed on 10 chromosomes. Phylogenetic analysis clearly showed that the family was classified as two clades: class I and class II. Gene structure and conserved motif analysis showed that the same clade genes have similar gene structures and conserved motifs. The expression profiles of 39 TCP genes were determined through quantitative real-time PCR. Most CIN-type BrrTCP genes were highly expressed in leaf. The members of CYC/TB1 subclade are highly expressed in flower bud and weakly expressed in root. By contrast, class I clade showed more widespread but less tissue-specific expression patterns. Yeast two-hybrid data show that BrrTCP proteins preferentially formed heterodimers. The function of BrrTCP2 was confirmed through ectopic expression of BrrTCP2 in wild-type and loss-of-function ortholog mutant of Arabidopsis. Overexpression of BrrTCP2 in wild-type Arabidopsis resulted in the diminished leaf size. Overexpression of BrrTCP2 in triple mutants of tcp2/4/10 restored the leaf phenotype of tcp2/4/10 to the phenotype of wild type. The comprehensive analysis of turnip TCP gene family provided the foundation to further study the roles of TCP genes in turnips.

  9. Molecular and Electrophysiological Characterization of GABAergic Interneurons Expressing the Transcription Factor COUP-TFII in the Adult Human Temporal Cortex

    Science.gov (United States)

    Varga, Csaba; Tamas, Gabor; Barzo, Pal; Olah, Szabolcs; Somogyi, Peter

    2015-01-01

    Transcription factors contribute to the differentiation of cortical neurons, orchestrate specific interneuronal circuits, and define synaptic relationships. We have investigated neurons expressing chicken ovalbumin upstream promoter transcription factor II (COUP-TFII), which plays a role in the migration of GABAergic neurons. Whole-cell, patch-clamp recording in vitro combined with colocalization of molecular cell markers in the adult cortex differentiates distinct interneurons. The majority of strongly COUP-TFII-expressing neurons were in layers I–III. Most calretinin (CR) and/or cholecystokinin- (CCK) and/or reelin-positive interneurons were also COUP-TFII-positive. CR-, CCK-, or reelin-positive neurons formed 80%, 20%, or 17% of COUP-TFII-positive interneurons, respectively. About half of COUP-TFII-/CCK-positive interneurons were CR-positive, a quarter of them reelin-positive, but none expressed both. Interneurons positive for COUP-TFII fired irregular, accommodating and adapting trains of action potentials (APs) and innervated mostly small dendritic shafts and rarely spines or somata. Paired recording showed that a calretinin-/COUP-TFII-positive interneuron elicited inhibitory postsynaptic potentials (IPSPs) in a reciprocally connected pyramidal cell. Calbindin, somatostatin, or parvalbumin-immunoreactive interneurons and most pyramidal cells express no immunohistochemically detectable COUP-TFII. In layers V and VI, some pyramidal cells expressed a low level of COUP-TFII in the nucleus. In conclusion, COUP-TFII is expressed in a diverse subset of GABAergic interneurons predominantly innervating small dendritic shafts originating from both interneurons and pyramidal cells. PMID:25787832

  10. Expression, purification, and characterization of a diabody against the most important angiogenesis cell receptor: Vascular endothelial growth factor receptor 2

    Directory of Open Access Journals (Sweden)

    Mahdi Behdani

    2012-01-01

    Full Text Available Antibodies and their derivative fragments have long been used as tools in a variety of applications, in fundamental research work, biotechnology, diagnosis, and therapy. Camels produce single heavy-chain antibodies (VHH in addition to usual antibodies. These minimal-sized binders are very robust and bind the antigen with high affinity in a monomeric state. Vascular endothelial growth factor recepror-2 (VEGFR2 is an important tumor-associated receptor that blockade of its signaling can lead to the inhibition of neovascularization and tumor metastasis. Here, we describe the construction, expression, and purification VEGFR2-specific Diabody. Two variable fragments of a same camel anti-VEGFR2 antibody were linked together by the upper hinge segment of antibody to make a diabody. We showed the ability of diabody to recognition of VEGFR2 on the cell surface by FACS. Diabodies can be produced in the low-cost prokaryotic expression system, so they are suitable molecules for diagnostic and therapeutic issues.

  11. Molecular Characterization of a Leaf Senescence-Related Transcription Factor BrWRKY75 of Chinese Flowering Cabbage

    Directory of Open Access Journals (Sweden)

    Xiaoli Tan

    2016-09-01

    Full Text Available WRKY is a plant-specific transcription factor (TF involved in the regulation of many biological processes; however, its role in leaf senescence of leafy vegetables remains unknown. In the present work, a WRKY TF, termed BrWRKY75 was isolated from Chinese flowering cabbage [Brassica rapa L. ssp. chinensis (L. Mokino var. utilis Tsen et Lee]. Analysis of deduced amino acid sequence and the phylogenetic tree showed that BrWRKY75 has high homology with WRKY75 from Brassica oleracea and Arabidopsis thaliana, and belongs to the II c sub-group. Sub-cellular localization and transcriptional activity analysis revealed that BrWRKY75 is a nuclear protein with transcriptional repression activity, and was up-regulated during leaf senescence. Electrophoretic mobility shift assay confirmed that BrWRKY75 directly bound to the W-box (TTGAC cis-element. Collectively, these results provide a basis for further investigation of the transcriptional regulation of Chinese flowering cabbage leaf senescence.

  12. Characterization of ApB73, a virulence factor important for colonization of Zea mays by the smut Ustilago maydis.

    Science.gov (United States)

    Stirnberg, Alexandra; Djamei, Armin

    2016-12-01

    The biotrophic fungus Ustilago maydis, the causal agent of corn smut disease, uses numerous small secreted effector proteins to suppress plant defence responses and reshape the host metabolism. However, the role of specific effectors remains poorly understood. Here, we describe the identification of ApB73 (Apathogenic in B73), an as yet uncharacterized protein essential for the successful colonization of maize by U. maydis. We show that apB73 is transcriptionally induced during the biotrophic stages of the fungal life cycle. The deletion of the apB73 gene results in cultivar-specific loss of gall formation in the host. The ApB73 protein is conserved among closely related smut fungi. However, using virulence assays, we show that only the orthologue of the maize-infecting head smut Sporisorium reilianum can complement the mutant phenotype of U. maydis. Although microscopy shows that ApB73 is secreted into the biotrophic interface, it seems to remain associated with fungal cell wall components or the fungal plasma membrane. Taken together, the results show that ApB73 is a conserved and important virulence factor of U. maydis that localizes to the interface between the pathogen and its host Zea mays. © 2016 THE AUTHORS. MOLECULAR PLANT PATHOLOGY PUBLISHED BY BRITISH SOCIETY FOR PLANT PATHOLOGY AND JOHN WILEY & SONS LTD.

  13. Molecular cloning, characterization, and expression of human ADP-ribosylation factors: Two guanine nucleotide-dependent activators of cholera toxin

    International Nuclear Information System (INIS)

    Bobak, D.A.; Nightingale, M.S.; Murtagh, J.J.; Price, S.R.; Moss, J.; Vaughan, M.

    1989-01-01

    ADP-ribosylation factors (ARFs) are small guanine nucleotide-binding proteins that enhance the enzymatic activities of cholera toxin. Two ARF cDNAs, ARF1 and ARF3, were cloned from a human cerebellum library. Based on deduced amino acid sequences and patterns of hybridization of cDNA and oligonucleotide probes with mammalian brain poly(A) + RNA, human ARF1 is the homologue of bovine ARF1. Human ARF3, which differs from bovine ARF1 and bovine ARF2, appears to represent a newly identified third type of ARF. Hybridization patterns of human ARF cDNA and clone-specific oligonucleotides with poly(A) + RNA are consistent with the presence of at least two, and perhaps four, separate ARF messages in human brain. In vitro translation of ARF1, ARF2, and ARF3 produced proteins that behaved, by SDS/PAGE, similar to a purified soluble brain ARF. Deduced amino acid sequences of human ARF1 and ARF3 contain regions, similar to those in other G proteins, that are believed to be involved in GTP binding and hydrolysis. ARFS also exhibit a modest degree of homology with a bovine phospholipase C. The observations reported here support the conclusion that the ARFs are members of a multigene family of small guanine nucleotide-binding proteins. Definition of the regulation of ARF mRNAs and of function(s) of recombinant ARF proteins will aid in the elucidation of the physiologic role(s) of ARFs

  14. Structural characterization of the α-mating factor prepro-peptide for secretion of recombinant proteins in Pichia pastoris.

    Science.gov (United States)

    Chahal, Sabreen; Wei, Peter; Moua, Pachai; Park, Sung Pil James; Kwon, Janet; Patel, Arth; Vu, Anthony T; Catolico, Jason A; Tsai, Yu Fang Tina; Shaheen, Nadia; Chu, Tiffany T; Tam, Vivian; Khan, Zill-E-Huma; Joo, Hyun Henry; Xue, Liang; Lin-Cereghino, Joan; Tsai, Jerry W; Lin-Cereghino, Geoff P

    2017-01-20

    The methylotrophic yeast Pichia pastoris has been used extensively for expressing recombinant proteins because it combines the ease of genetic manipulation, the ability to provide complex posttranslational modifications and the capacity for efficient protein secretion. The most successful and commonly used secretion signal leader in Pichia pastoris has been the alpha mating factor (MATα) prepro secretion signal. However, limitations exist as some proteins cannot be secreted efficiently, leading to strategies to enhance secretion efficiency by modifying the secretion signal leader. Based on a Jpred secondary structure prediction and knob-socket modeling of tertiary structure, numerous deletions and duplications of the MATα prepro leader were engineered to evaluate the correlation between predicted secondary structure and the secretion level of the reporters horseradish peroxidase (HRP) and Candida antarctica lipase B. In addition, circular dichroism analyses were completed for the wild type and several mutant pro-peptides to evaluate actual differences in secondary structure. The results lead to a new model of MATα pro-peptide signal leader, which suggests that the N and C-termini of MATα pro-peptide need to be presented in a specific orientation for proper interaction with the cellular secretion machinery and for efficient protein secretion. Copyright © 2016 Elsevier B.V. All rights reserved.

  15. Genome-Wide Characterization and Expression Profiling of the AUXIN RESPONSE FACTOR (ARF) Gene Family in Eucalyptus grandis

    Science.gov (United States)

    Yu, Hong; Soler, Marçal; Mila, Isabelle; San Clemente, Hélène; Savelli, Bruno; Dunand, Christophe; Paiva, Jorge A. P.; Myburg, Alexander A.; Bouzayen, Mondher; Grima-Pettenati, Jacqueline; Cassan-Wang, Hua

    2014-01-01

    Auxin is a central hormone involved in a wide range of developmental processes including the specification of vascular stem cells. Auxin Response Factors (ARF) are important actors of the auxin signalling pathway, regulating the transcription of auxin-responsive genes through direct binding to their promoters. The recent availability of the Eucalyptus grandis genome sequence allowed us to examine the characteristics and evolutionary history of this gene family in a woody plant of high economic importance. With 17 members, the E. grandis ARF gene family is slightly contracted, as compared to those of most angiosperms studied hitherto, lacking traces of duplication events. In silico analysis of alternative transcripts and gene truncation suggested that these two mechanisms were preeminent in shaping the functional diversity of the ARF family in Eucalyptus. Comparative phylogenetic analyses with genomes of other taxonomic lineages revealed the presence of a new ARF clade found preferentially in woody and/or perennial plants. High-throughput expression profiling among different organs and tissues and in response to environmental cues highlighted genes expressed in vascular cambium and/or developing xylem, responding dynamically to various environmental stimuli. Finally, this study allowed identification of three ARF candidates potentially involved in the auxin-regulated transcriptional program underlying wood formation. PMID:25269088

  16. Reserve, thin form-factor, hypochlorite-based cells for powering portable systems: Manufacture (including MEMS processes), performance and characterization

    Energy Technology Data Exchange (ETDEWEB)

    Cardenas-Valencia, Andres M.; Langebrake, Larry [Center for Ocean Technology, University of South Florida, 140 Seventh Ave. S., St. Petersburg, FL (United States); Biver, Carl J. [Center for Ocean Technology, University of South Florida, 140 Seventh Ave. S., St. Petersburg, FL (United States); Department of Chemical Engineering, University of South Florida, 4202 E. Fowler Ave. Tampa, FL (United States)

    2007-03-30

    This work focuses on fabrication routes and performance evaluation of thin form-factors, reserve cells, as a powering alternative for expendable and/or remotely operated systems. The catalytic decomposition of sodium hypochlorite solutions is revisited herein with two cost-effective anodes: zinc and aluminum. Aluminum, even though the most expensive of the utilized anodes, constituted cells with double the energy content (up to 55 Wh kg{sup -1}) than those fabricated with zinc. Even though the hypochlorite concentration in the solution limits the cells' operational life, attractive performances (1.0 V with a current of 10 mA) for the manufactured cells are obtained. It is shown that micro fabrication processes, allowing for close electrodes interspacing, provided high faradic and columbic efficiencies of up to 70 and 100%, respectively. Obtained specific energies (50-120 Wh kg{sup -1}) are in the same order of magnitude than batteries currently used for powering deployable systems. Experimental results show that a simple model that linearly relates over potentials and the electrical load, adequately describe all the cell designs. A mathematical model based on a kinetic-mechanistic scheme that relates the current output as a function of time agrees fairly well with results obtained activating cells with various concentrations of NaOCl solutions. (author)

  17. Characterization of an insulin-like growth factor-I/somatomedin-C radioimmunoassay specific for the C-peptide region

    International Nuclear Information System (INIS)

    Hintz, R.L.; Liu, F.; Seegan, G.

    1982-01-01

    Insulin-like growth factor-I (IGF-I) and somatomedin-C (SM-C) have been shown to be functionally identical by a number of criteria. We have synthesized the 12 amino acid C-peptide region of IGF-I (Gly-Tyr-Gly-Ser-Ser-Ser-Arg-Arg-Ala-Pro-Glu-Thr) and developed a RIA based on antibodies against this synthetic peptide. IGF-I and SM-C were indistinguishable in this RIA. No other peptides competed for this antiserum. The SM-C/IGF-I values of acid-chromatographed serum were strongly age dependent. The mean of children 1-5 yr old was 0.67 +/- 0.033 U/ml (mean +/- sD; n = 23), whereas the mean of children 12-17 yr old was 2.01 +/- 0.66 U/ml (n = 39) and the mean of 38 adults 26-85 yr old was 1.05 +/- 0.34. The SM-C/IGF-I values measured by this RIA were also growth hormone dependent. Thus, this region-specific RIA provides a clinically useful assessment of serum SM-C/IGF-I levels

  18. Prospective Characterization of the Risk Factors for Transmission and Symptoms of Primary Human Herpesvirus Infections Among Ugandan Infants.

    Science.gov (United States)

    Gantt, Soren; Orem, Jackson; Krantz, Elizabeth M; Morrow, Rhoda Ashley; Selke, Stacy; Huang, Meei-Li; Schiffer, Joshua T; Jerome, Keith R; Nakaganda, Annet; Wald, Anna; Casper, Corey; Corey, Lawrence

    2016-07-01

    Human herpesvirus (HHV) infections are common during infancy. Primary infections are frequently asymptomatic and best studied prospectively by using direct viral detection. Oropharyngeal swab specimens were collected weekly from Ugandan newborn infants, their mothers, and other children in the household. Blood specimens were collected every 4 months. Samples were tested for herpes simplex virus (HSV) types 1 and 2, Epstein-Barr virus (EBV), cytomegalovirus (CMV), HHV-6A, HHV-6B, and HHV-8, using quantitative polymerase chain reaction. Thirty-two infants, 32 mothers, and 49 other household children were followed for a median of 57 weeks. Seventeen mothers had human immunodeficiency virus type 1 (HIV) infection; no infants acquired HIV-1. The 12-month incidence of postnatal infection was 76% for HHV-6B, 59% for CMV, 47% for EBV, 8% for HSV-1, and 0% for HHV-8. The quantity of oropharyngeal shedding by contacts was associated with HHV-6A or HHV-6B transmission. Maternal HIV-1 infection was associated with EBV transmission, while breastfeeding and younger child contacts were associated with CMV transmission. Except for HSV-1, primary HHV infections were subclinical. By capturing exposures and acquisition events, we found that the incidence and risk factors of infection vary by HHV type. HSV-1 infection, unlike other HHV infections, caused acute clinical illness in these infants. © The Author 2016. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail journals.permissions@oup.com.

  19. Genome-wide characterization and expression profiling of the AUXIN RESPONSE FACTOR (ARF gene family in Eucalyptus grandis.

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    Hong Yu

    Full Text Available Auxin is a central hormone involved in a wide range of developmental processes including the specification of vascular stem cells. Auxin Response Factors (ARF are important actors of the auxin signalling pathway, regulating the transcription of auxin-responsive genes through direct binding to their promoters. The recent availability of the Eucalyptus grandis genome sequence allowed us to examine the characteristics and evolutionary history of this gene family in a woody plant of high economic importance. With 17 members, the E. grandis ARF gene family is slightly contracted, as compared to those of most angiosperms studied hitherto, lacking traces of duplication events. In silico analysis of alternative transcripts and gene truncation suggested that these two mechanisms were preeminent in shaping the functional diversity of the ARF family in Eucalyptus. Comparative phylogenetic analyses with genomes of other taxonomic lineages revealed the presence of a new ARF clade found preferentially in woody and/or perennial plants. High-throughput expression profiling among different organs and tissues and in response to environmental cues highlighted genes expressed in vascular cambium and/or developing xylem, responding dynamically to various environmental stimuli. Finally, this study allowed identification of three ARF candidates potentially involved in the auxin-regulated transcriptional program underlying wood formation.

  20. Characterizing fluorescent dissolved organic matter in a membrane bioreactor via excitation-emission matrix combined with parallel factor analysis.

    Science.gov (United States)

    Maqbool, Tahir; Quang, Viet Ly; Cho, Jinwoo; Hur, Jin

    2016-06-01

    In this study, we successfully tracked the dynamic changes in different constitutes of bound extracellular polymeric substances (bEPS), soluble microbial products (SMP), and permeate during the operation of bench scale membrane bioreactors (MBRs) via fluorescence excitation-emission matrix (EEM) combined with parallel factor analysis (PARAFAC). Three fluorescent groups were identified, including two protein-like (tryptophan-like C1 and tyrosine-like C2) and one microbial humic-like components (C3). In bEPS, protein-like components were consistently more dominant than C3 during the MBR operation, while their relative abundance in SMP depended on aeration intensities. C1 of bEPS exhibited a linear correlation (R(2)=0.738; pbEPS amounts in sludge, and C2 was closely related to the stability of sludge. The protein-like components were more greatly responsible for membrane fouling. Our study suggests that EEM-PARAFAC can be a promising monitoring tool to provide further insight into process evaluation and membrane fouling during MBR operation. Copyright © 2016 Elsevier Ltd. All rights reserved.

  1. Characterizing abnormal behavior in a large population of zoo-housed chimpanzees: prevalence and potential influencing factors

    Directory of Open Access Journals (Sweden)

    Sarah L. Jacobson

    2016-07-01

    Full Text Available Abnormal behaviors in captive animals are generally defined as behaviors that are atypical for the species and are often considered to be indicators of poor welfare. Although some abnormal behaviors have been empirically linked to conditions related to elevated stress and compromised welfare in primates, others have little or no evidence on which to base such a relationship. The objective of this study was to investigate a recent claim that abnormal behavior is endemic in the captive population by surveying a broad sample of chimpanzees (Pan troglodytes, while also considering factors associated with the origins of these behaviors. We surveyed animal care staff from 26 accredited zoos to assess the prevalence of abnormal behavior in a large sample of chimpanzees in the United States for which we had information on origin and rearing history. Our results demonstrated that 64% of this sample was reported to engage in some form of abnormal behavior in the past two years and 48% of chimpanzees engaged in abnormal behavior other than coprophagy. Logistic regression models were used to analyze the historical variables that best predicted the occurrence of all abnormal behavior, any abnormal behavior that was not coprophagy, and coprophagy. Rearing had opposing effects on the occurrence of coprophagy and the other abnormal behaviors such that mother-reared individuals were more likely to perform coprophagy, whereas non-mother-reared individuals were more likely to perform other abnormal behaviors. These results support the assertion that coprophagy may be classified separately when assessing abnormal behavior and the welfare of captive chimpanzees. This robust evaluation of the prevalence of abnormal behavior in our sample from the U.S. zoo population also demonstrates the importance of considering the contribution of historical variables to present behavior, in order to better understand the causes of these behaviors and any potential relationship to

  2. Persistent Postoperative Pain after Cardiac Surgery: Incidence, Characterization, Associated Factors and its impact in Quality of Life.

    Science.gov (United States)

    Guimarães-Pereira, Luís; Farinha, Filomena; Azevedo, Luís; Abelha, Fernando; Castro-Lopes, José

    2016-10-01

    Cardiac surgery (CS) ranks among the most frequently performed interventions worldwide and persistent postoperative pain (PPP) has been recognized as a relevant clinical outcome in this context. We aimed to evaluate its incidence, characteristics, associated factors and patient's quality of life (QoL). Observational prospective study conducted in patients undergoing CS in a tertiary university hospital. PPP was defined as persistent pain after surgery with higher than 3 months' duration, after excluding other causes of pain. We used a set of questionnaires for data collection: Pain Catastrophizing Scale, Duke Health Profile, Brief Pain Inventory Short Form, McGill Pain Questionnaire Short Form, Douleur Neuropathique en 4 Questions and standardized questions regarding pain periodicity. A total of 288 patients have completed the study and 43% presented PPP assessed at 3 months (PPP3M); out of which 84% were not under any treatment. PPP patients reported significantly lower QoL, and a neuropathic pain (NP) component was present in 50% of them. Younger age, female gender, higher body mass index, catastrophizing, coronary artery bypass graft, osteoarthritis, history of previous surgery (excluding sternotomy) and moderate to severe acute postoperative pain were independent predictors of PPP3M. This is the first study comprehensively describing PPP after CS and identifying NP in half of them. Our results support the important role that PPP plays after CS in considering its interference in patients' daily life and their lower QoL, which deserves the attention of health care professionals in order to improve prevention, assessment and treatment of these patients. WHAT DOES THIS STUDY ADD?: This study comprehensively describes persistent postoperative pain (PPP) after cardiac surgery (CS) and identifies neuropathic pain (NP) in half of them. Our results support the important role that PPP plays after CS in considering its interference in patients' daily life and their

  3. Partial characterization of insulin-like growth factor I in primary human lung cancers using immunohistochemical and receptor autoradiographic techniques

    International Nuclear Information System (INIS)

    Shigematsu, K.; Kataoka, Y.; Kamio, T.; Kurihara, M.; Niwa, M.; Tsuchiyama, H.

    1990-01-01

    We investigated primary human lung cancers resected surgically or obtained at autopsy. Included were squamous cell carcinoma (SQC) (five cases), adenocarcinoma (ADC) (six cases), large cell carcinoma (LCC) (four cases), and small cell carcinoma (SCC) (two cases). The objective of the study was to search for the presence of insulin-like growth factor I (IGF-I)-like immunoreactivity using immunohistochemical staining and for the localization of IGF-I binding sites, using in vitro quantitative receptor autoradiographic techniques. IGF-I-like immunostaining was present in all cases of SQC, ADC, and LCC, but not in cases of SCC. Strong immunostaining was observed in cases of SQC. On the other hand, ADC and LCC tissues showed a moderate or weak staining. Specific binding sites for IGF-I were present in all cases of SQC, ADC, LCC, and SCC examined. High densities of 125I-IGF-I binding sites were localized in cases of SQC and SCC. Low to high densities of the binding sites were found in LCC. Cases of ADC showed low densities of 125I-IGF-I binding sites. Specific binding obtained at a concentration of 80 pM 125I-IGF-I was competitively displaced by unlabeled IGF-I, with a 50% inhibitory concentration value of 1.84 +/- 0.31 x 10(-10) mol, whereas human insulin was much less potent in displacing the binding. This specificity profile is consistent with characteristics of IGF-I receptors. Scatchard analysis showed the presence of a single class of high affinity binding sites for IGF-I, with a Kd of approximately 1 nmol. Thus, the possibility that IGF-I may play a role in the growth of human lung cancers would have to be considered

  4. Expression of Tumor Necrosis Factor Receptor 2 Characterizes TLR9-Driven Formation of Interleukin-10-Producing B Cells

    Directory of Open Access Journals (Sweden)

    Olga Ticha

    2018-01-01

    Full Text Available B cell-derived interleukin-10 (IL-10 production has been described as a hallmark for regulatory function in B lymphocytes. However, there is an ongoing debate on the origin of IL-10-secreting B cells and lack of specific surface markers has turned into an important obstacle for studying human B regulatory cells. In this study, we propose that tumor necrosis factor receptor 2 (TNFR2 expression can be used for enrichment of IL-10-secreting B cells. Our data confirm that IL-10 production can be induced by TLR9 stimulation with CpG ODN and that IL-10 secretion accompanies differentiation of peripheral blood B cells into plasma blasts. We further show that CpG ODN stimulation induces TNFR2 expression, which correlates with IL-10 secretion and terminal differentiation. Indeed, flow cytometric sorting of TNFR2+ B cells revealed that TNFR2+ and TNFR2− fractions correspond to IL-10+ and IL-10− fractions, respectively. Furthermore, CpG-induced TNFR2+ B cells were predominantly found in the IgM+ CD27+ B cell subset and spontaneously released immunoglobulin. Finally, our data corroborate the functional impact of TNFR2 by demonstrating that stimulation with a TNFR2 agonist significantly augments IL-10 and IL-6 production in B cells. Altogether, our data highlight a new role for TNFR2 in IL-10-secreting human B lymphocytes along with the potential to exploit this finding for sorting and isolation of this currently ill-defined B cell subset.

  5. Characterization of a novel anti-lipopolysaccharide factor isoform (SpALF5) in mud crab, Scylla paramamosain.

    Science.gov (United States)

    Sun, Wanwei; Wan, Weisong; Zhu, Shuo; Wang, Shasha; Wang, Shuqi; Wen, Xiaobo; Zheng, Huaiping; Zhang, Yueling; Li, Shengkang

    2015-04-01

    Anti-lipopolysaccharide factors (ALFs), the potential antimicrobial peptides that bind and neutralize lipopolysaccharide (LPS), are common effectors of innate immunity in crustaceans. In this study, a novel isoform of ALFs (SpALF5) was isolated from the hemocytes of mud crab Scylla paramamosain. The full-length 975bp SpALF5 contains a 375bp open reading frame (ORF) encoding 125 amino acids. Although SpALF5 exhibits a low degree of nucleotide homology with other reported ALFs, it contains the conserved amino acid sequence with a signal peptide and a LPS-binding domain including two conservative cysteine residues. The genomic organization of SpALF5 consists of four exons and three introns, with each intron containing one or more tandem repeats. Unlike most of ALFs mainly distributed in crab hemocytes, SpALF5 transcript was predominantly observed in the brain, muscle and skin, while barely detected in the hemocytes in our study. In situ hybridization assay also showed that SpALF5 mRNA was localized in brain, muscle and skin tissues of mud crab. Further, SpALF5 transcript was significantly up-regulated after challenge with LPS, polyinosinic polycytidylic acid (PolyI:C) (with the except of that in brain), Vibrio parahemolyticus or white spot syndrome virus (WSSV). The recombinant SpALF5 protein showed a varying degree of binding activity towards bacteria and fungus. Moreover, in vitro, the recombinant SpALF5 revealed a strong antimicrobial activity against Gram-negative bacteria (V. parahemolyticus, Vibrio alginolyticus, Escherichia coli, Aeromonas hydrophila) and fungus (Sacchromyces cerevisiae), but could only inhibited the growth of some Gram-positive bacteria like Staphylococcus aureus. The results suggest that SpALF5 is a potent immune protector and plays an important role in immune defense against invading pathogens in S. paramamosain. Copyright © 2014 Elsevier Ltd. All rights reserved.

  6. Wound induced tanscriptional regulation of benzylisoquinoline pathway and characterization of wound inducible PsWRKY transcription factor from Papaver somniferum.

    Directory of Open Access Journals (Sweden)

    Sonal Mishra

    Full Text Available Wounding is required to be made in the walls of the green seed pod of Opium poppy prior exudation of latex. To withstand this kind of trauma plants regulate expression of some metabolites through an induced transcript level. 167 unique wound-inducible ESTs were identified by a repetitive round of cDNA subtraction after 5 hours of wounding in Papaver somniferum seedlings. Further repetitive reverse northern analysis of these ESTs revealed 80 transcripts showing more than two fold induction, validated through semi-quantitative RT-PCR & real time expression analysis. One of the major classified categories among identified ESTs belonged to benzylisoquinoline transcripts. Tissue specific metabolite analysis of benzylisoquinoline alkaloids (BIAs in response to wounding revealed increased accumulation of narcotine and papaverine. Promoter analysis of seven transcripts of BIAs pathway showed the presence of W-box cis-element with the consensus sequence of TGAC, which is the proposed binding site for WRKY type transcription factors. One of the Wound inducible 'WRKY' EST isolated from our subtracted library was made full-length and named as 'PsWRKY'. Bacterially expressed PsWRKY interacted with the W-box element having consensus sequence TTGACT/C present in the promoter region of BIAs biosynthetic pathway genes. PsWRKY further activated the TYDC promoter in yeast and transiently in tobacco BY2 cells. Preferential expression of PsWRKY in straw and capsule and its interaction with consensus W-box element present in BIAs pathway gene transcripts suggest its possible involvement in the wound induced regulation of BIAs pathway.

  7. Chemical characterization of PM1.0 aerosol in Delhi and source apportionment using positive matrix factorization.

    Science.gov (United States)

    Jaiprakash; Singhai, Amrita; Habib, Gazala; Raman, Ramya Sunder; Gupta, Tarun

    2017-01-01

    Fine aerosol fraction (particulate matter with aerodynamic diameter <= 1.0 μm (PM) 1.0 ) over the Indian Institute of Technology Delhi campus was monitored day and night (10 h each) at 30 m height from November 2009 to March 2010. The samples were analyzed for 5 ions (NH 4 + , NO 3 - , SO 4 2- , F - , and Cl - ) and 12 trace elements (Na, K, Mg, Ca, Pb, Zn, Fe, Mn, Cu, Cd, Cr, and Ni). Importantly, secondary aerosol (sulfate and nitrate) formation was observed during dense foggy events, supporting the fog-smog-fog cycle. A total of 76 samples were used for source apportionment of PM mass. Six factors were resolved by PMF analyses and were identified as secondary aerosol, secondary chloride, biomass burning, soil dust, iron-rich source, and vehicular emission. The geographical location of the sources and/or preferred transport pathways was identified by conditional probability function (for local sources) and potential source contribution function (for regional sources) analyses. Medium- and small-scale metal processing (e.g. steel sheet rolling) industries in Haryana and National Capital Region (NCR) Delhi, coke and petroleum refining in Punjab, and thermal power plants in Pakistan, Punjab, and NCR Delhi were likely contributors to secondary sulfate, nitrate, and secondary chloride at the receptor site. The agricultural residue burning after harvesting season (Sept-Dec and Feb-Apr) in Punjab, and Haryana contributed to potassium at receptor site during November-December and March 2010. The soil dust from North and East Pakistan, and Rajasthan, North-East Punjab, and Haryana along with the local dust contributed to soil dust at the receptor site, during February and March 2010. A combination of temporal behavior and air parcel trajectory ensemble analyses indicated that the iron-rich source was most likely a local source attributed to emissions from metal processing facilities. Further, as expected, the vehicular emissions source did not show any seasonality and

  8. Characterizing China's energy consumption with selective economic factors and energy-resource endowment: a spatial econometric approach

    Science.gov (United States)

    Jiang, Lei; Ji, Minhe; Bai, Ling

    2015-06-01

    Coupled with intricate regional interactions, the provincial disparity of energy-resource endowment and other economic conditions in China have created spatially complex energy consumption patterns that require analyses beyond the traditional ones. To distill the spatial effect out of the resource and economic factors on China's energy consumption, this study recast the traditional econometric model in a spatial context. Several analytic steps were taken to reveal different aspects of the issue. Per capita energy consumption (AVEC) at the provincial level was first mapped to reveal spatial clusters of high energy consumption being located in either well developed or energy resourceful regions. This visual spatial autocorrelation pattern of AVEC was quantitatively tested to confirm its existence among Chinese provinces. A Moran scatterplot was employed to further display a relatively centralized trend occurring in those provinces that had parallel AVEC, revealing a spatial structure with attraction among high-high or low-low regions and repellency among high-low or low-high regions. By a comparison between the ordinary least square (OLS) model and its spatial econometric counterparts, a spatial error model (SEM) was selected to analyze the impact of major economic determinants on AVEC. While the analytic results revealed a significant positive correlation between AVEC and economic development, other determinants showed some intricate influential patterns. The provinces endowed with rich energy reserves were inclined to consume much more energy than those otherwise, whereas changing the economic structure by increasing the proportion of secondary and tertiary industries also tended to consume more energy. Both situations seem to underpin the fact that these provinces were largely trapped in the economies that were supported by technologies of low energy efficiency during the period, while other parts of the country were rapidly modernized by adopting advanced

  9. Item response theory and factor analysis as a mean to characterize occurrence of response shift in a longitudinal quality of life study in breast cancer patients

    Science.gov (United States)

    2014-01-01

    Background The occurrence of response shift (RS) in longitudinal health-related quality of life (HRQoL) studies, reflecting patient adaptation to disease, has already been demonstrated. Several methods have been developed to detect the three different types of response shift (RS), i.e. recalibration RS, 2) reprioritization RS, and 3) reconceptualization RS. We investigated two complementary methods that characterize the occurrence of RS: factor analysis, comprising Principal Component Analysis (PCA) and Multiple Correspondence Analysis (MCA), and a method of Item Response Theory (IRT). Methods Breast cancer patients (n = 381) completed the EORTC QLQ-C30 and EORTC QLQ-BR23 questionnaires at baseline, immediately following surgery, and three and six months after surgery, according to the “then-test/post-test” design. Recalibration was explored using MCA and a model of IRT, called the Linear Logistic Model with Relaxed Assumptions (LLRA) using the then-test method. Principal Component Analysis (PCA) was used to explore reconceptualization and reprioritization. Results MCA highlighted the main profiles of recalibration: patients with high HRQoL level report a slightly worse HRQoL level retrospectively and vice versa. The LLRA model indicated a downward or upward recalibration for each dimension. At six months, the recalibration effect was statistically significant for 11/22 dimensions of the QLQ-C30 and BR23 according to the LLRA model (p ≤ 0.001). Regarding the QLQ-C30, PCA indicated a reprioritization of symptom scales and reconceptualization via an increased correlation between functional scales. Conclusions Our findings demonstrate the usefulness of these analyses in characterizing the occurrence of RS. MCA and IRT model had convergent results with then-test method to characterize recalibration component of RS. PCA is an indirect method in investigating the reprioritization and reconceptualization components of RS. PMID:24606836

  10. p55-hGRF, a short natural form of the Ras-GDP exchange factor high yield production and characterization.

    Science.gov (United States)

    Meyer, P; Janin, J; Baudet-Nessler, S

    1999-08-01

    p55-hGRF, a natural short form of the guanine-nucleotide-releasing factor for p21-Ras from human brain, was expressed at high level in Escherichia coli as well as an engineered truncated form, p39-hGRF. A T7 polymerase expression system was used, resulting in the formation of insoluble cytoplasmic protein aggregates. The recombinant products were resolubilized, renatured and purified to homogeneity. The exchange activity of the refolded hGRF samples on H-Ras was comparable with that published for the soluble catalytic domain of the mouse counterpart, CDC25 Mm. Both p55-hGRF and p39-hGRF form dimers. We established a procedure to prepare and purify the complex with Ras. The results of the characterization study are consistent with a stoichiometry of 1:1 and an equilibrium between dimeric and monomeric forms of the complex.

  11. Genome-wide identification and characterization of WRKY transcriptional factor family in apple and analysis of their responses to waterlogging and drought stress.

    Science.gov (United States)

    Meng, Dong; Li, Yuanyuan; Bai, Yang; Li, Mingjun; Cheng, Lailiang

    2016-06-01

    As one of the largest transcriptional factor families in plants, WRKY genes play significant roles in various biotic and abiotic stress responses. Although the WRKY gene family has been characterized in a few plant species, the details remain largely unknown in the apple (Malus domestica Borkh.). In this study, we identified a total of 127 MdWRKYs from the apple genome, which were divided into four subgroups according to the WRKY domains and zinc finger motif. Most of them were mapped onto the apple's 17 chromosomes and were expressed in more than one tissue, including shoot tips, mature leaves, fruit and apple calli. We then contrasted WRKY expression patterns between calli grown in solid medium (control) and liquid medium (representing waterlogging stress) and found that 34 WRKY genes were differentially expressed between the two growing conditions. Finally, we determined the expression patterns of 10 selected WRKY genes in an apple rootstock, G41, in response to waterlogging and drought stress, which identified candidate genes involved in responses to water stress for functional analysis. Our data provide interesting candidate MdWRKYs for future functional analysis and demonstrate that apple callus is a useful system for characterizing gene expression and function in apple. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

  12. Identification of stable cytotoxic factors in the gas phase extract of cigarette smoke and pharmacological characterization of their cytotoxicity.

    Science.gov (United States)

    Noya, Yoichi; Seki, Koh-Ichi; Asano, Hiroshi; Mai, Yosuke; Horinouchi, Takahiro; Higashi, Tsunehito; Terada, Koji; Hatate, Chizuru; Hoshi, Akimasa; Nepal, Prabha; Horiguchi, Mika; Kuge, Yuji; Miwa, Soichi

    2013-12-06

    Smoking is a major risk factor for atherosclerotic vascular diseases, but the mechanism for its genesis is unknown. We have recently shown that the gas phase of cigarette smoke (nicotine- and tar-free cigarette smoke extract; CSE) likely to reach the systemic circulation contains stable substances which cause cytotoxicity like plasma membrane damage and cell death in cultured cells, and also that the plasma membrane damage is caused through sequential activation of protein kinase C (PKC) and NADPH oxidase (NOX) and the resulting generation of reactive oxygen species (PKC/NOX-dependent mechanism), whereas cell death is caused through PKC/NOX-dependent and -independent mechanisms. To identify these stable substances, the CSE was prepared by passing the main-stream smoke of 10 cigarettes through a Cambridge glass fiber filter, trapping of the smoke in a vessel cooled at -80°C, and subsequent dissolution in 10ml of water. The CSE was fractionated into nine fractions using reversed-phase HPLC, and each fraction was screened for cytotoxicity in cultured cells, using propidium iodide uptake assay for cell membrane damage and MTS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium] reduction assay for cell viability. The cytotoxicity was positive in two of the nine fractions (Fr2 and Fr5). After extraction of the active fractions into dichloromethane, GC/MS analysis identified 2-cyclopenten-1-one (CPO) in Fr5 but none in Fr2. After derivatization of the active fractions with O-(2,3,4,5,6-pentafluorobenzyl) hydroxylamine hydrochloride, GC/MS analysis identified acrolein, acetone and propionaldehyde in Fr2, and methyl vinyl ketone (MVK) in Fr5. After 4-h incubation, authentic acrolein and MVK induced concentration-dependent cytotoxicity with EC50 values of 75.9±8.2 and 47.0±8.0μM (mean±SEM; n=3), respectively, whereas acetone, propionaldehyde and CPO were without effect. However, after 24-h incubation, CPO induced concentration

  13. Characterization of novel peptide-specific antibodies against the translation elongation factor eEF1A2 and their application for cancer research

    Directory of Open Access Journals (Sweden)

    Shalak V. F.

    2014-11-01

    Full Text Available Aim. We intend to characterize the new peptide-specific antibodies against the isoform 2 of translation elongation factor 1A (eEF1A2 and determine its presence in the postoperative samples of human breast, lung and stomach tumor tissues. Methods. The analysis of antibody specificity was performed by enzyme-linked immunosorbent assay, immunoblotting and immunohistochemistry. Immunoblotting and immunohistochemistry were used for the determination of the eEF1A2 in the human tumor samples, as well as in the samples of normal tissues surrounding tumors. Results. The antibodies obtained against the eEF1A2 specifically recognized this protein in the cell extracts and histological sections and did not cross-react with the elongation factor 1A isoform 1. eEF1A2 was revealed in the postoperative samples of breast, lung and stomach tumors as well as in the putative normal tissues surrounding tumors. Conclusions. The antibodies obtained against eEF1A2 are highly specific for the antigen and can be used for the immunological studies of tumors.

  14. Characterization of CgHIFα-Like, a Novel bHLH-PAS Transcription Factor Family Member, and Its Role under Hypoxia Stress in the Pacific Oyster Crassostrea gigas.

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    Ting Wang

    Full Text Available Hypoxia-inducible factor (HIF, a critical member of the basic-helix-loop-helix (bHLH-containing Per-Arnt-Sim (PAS protein family, is a master transcription factor involved in maintaining oxygen homeostasis. In the present study, we isolated and characterized a novel bHLH-PAS family member, CgHIFα-like gene, from the Pacific oyster Crassostrea gigas, and determined its importance during hypoxia stress. The 3020-bp CgHIFα-like cDNA encoded a protein of 888 amino acids. The predicted CgHIFα-like amino acid sequence was conserved in the N-terminal bHLH, PAS, and PAC domains (but not in the C-terminal domain and was most closely related to the HIF family in the bHLH-PAS protein phylogenic tree. Similar to the mammalian HIF-1α, CgHIFα-like could be expressed as four mRNA isoforms containing alternative 5'-untranslated regions and different translation initiation codons. At the mRNA level, these isoforms were expressed in a tissue-specific manner and showed increased transcription to varying degrees under hypoxic conditions. Additionally, the western blot analysis demonstrated that CgHIFα-like was induced by hypoxia. Electrophoretic mobility shift assay indicated that CgHIFα-like could bind to the hypoxia responsive element (HRE, whereas dual-luciferase reporter analysis demonstrated that CgHIFα-like could transactivate the reporter gene containing the HREs. In addition to CgHIFα-like, we identified CgARNT from the C. gigas, analyzed its expression pattern, and confirmed its interaction with CgHIFα-like using a yeast two-hybrid assay. In conclusion, this is the first report on the cloning and characterization of a novel hypoxia transcription factor in mollusks, which could accumulate under hypoxia and regulate hypoxia related gene expression by binding to HRE and dimerizing with CgARNT. As only one member of HIF has been identified in invertebrates to date, our results provide new insights into the unique mechanisms of hypoxia tolerance in

  15. Molecular Characterization and Expression Analysis of Insulin-like Growth Factor-1 and Insulin-like Growth Factor Binding Protein-1 Genes in Qinghai-Tibet Plateau and Lowland

    Directory of Open Access Journals (Sweden)

    Ya-bing Chen

    2015-01-01

    Full Text Available Insulin-like growth factor-1 (IGF-1 and insulin-like growth factor binding protein-1 (IGFBP-1 play a pivotal role in regulating cellular hypoxic response. In this study, we cloned and characterized the genes encoding IGF-1 and IGFBP-1 to improve the current knowledge on their roles in highland Bos grunniens (Yak. We also compared their expression levels in the liver and kidney tissues between yaks and lowland cattle. We obtained full-length 465 bp IGF-1 and 792 bp IGFBP-1, encoding 154 amino acids (AA IGF-1, and 263 AA IGFBP-1 protein, respectively using reverse transcriptase-polyerase chain reaction (RT-PCR technology. Analysis of their corresponding amino acid sequences showed a high identity between B. grunniens and lowland mammals. Moreover, the two genes were proved to be widely distributed in the examined tissues through expression pattern analysis. Real-time PCR results revealed that IGF-1 expression was higher in the liver and kidney tissues in B. grunniens than in Bos taurus (p<0.05. The IGFBP-1 gene was expressed at a higher level in the liver (p<0.05 of B. taurus than B. grunniens, but it has a similar expression level in the kidneys of the two species. These results indicated that upregulated IGF-1 and downregulated IGFBP-1 are associated with hypoxia adaptive response in B. grunniens.

  16. Generation and characterization of tabalumab, a human monoclonal antibody that neutralizes both soluble and membrane-bound B-cell activating factor

    Directory of Open Access Journals (Sweden)

    Manetta J

    2014-08-01

    Full Text Available Joseph Manetta, Holly Bina, Paul Ryan, Niles Fox, Derrick R Witcher, Kristine Kikly Biotechnology Discovery Research, Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN, USA Abstract: B-cell activating factor (BAFF is a B-cell survival factor with a key role in B-cell homeostasis and tolerance. Dysregulated BAFF expression may contribute to autoimmune diseases or B-cell malignancies via effects on abnormal B-lymphocyte activation, proliferation, survival, and immunoglobulin secretion. Monoclonal antibodies were generated against human BAFF, characterized for species specificity and affinity, and screened for the ability to neutralize both membrane-bound and soluble BAFF. In addition, studies were undertaken to determine the relative potency of membrane-bound and soluble BAFF. Tabalumab has a high affinity for human, cynomolgus monkey, and rabbit BAFF. No binding to mouse BAFF was detected. Tabalumab was able to neutralize soluble human, cynomolgus monkey, or rabbit BAFF with equal potency. Our data demonstrate that membrane-bound BAFF can be a more potent stimulus for B-cells than soluble BAFF, and tabalumab also neutralized membrane-bound BAFF. Tabalumab prevented BAFF from binding to BAFF receptors and demonstrated pharmacodynamic effects in human BAFF transgenic mice. Tabalumab is a high-affinity human antibody with neutralizing activity against membrane-bound and soluble BAFF. Given our findings that membrane-bound BAFF can have greater in vitro potency than soluble BAFF, neutralization of both forms of BAFF is likely to be important for optimal therapeutic effect. Keywords: autoimmunity, B-cell malignancies, B-cell survival factor, BAFF

  17. Characterization and functional analysis of hypoxia-inducible factor HIF1α and its inhibitor HIF1αn in tilapia.

    Science.gov (United States)

    Li, Hong Lian; Gu, Xiao Hui; Li, Bi Jun; Chen, Xiao; Lin, Hao Ran; Xia, Jun Hong

    2017-01-01

    Hypoxia is a major cause of fish morbidity and mortality in the aquatic environment. Hypoxia-inducible factors are very important modulators in the transcriptional response to hypoxic stress. In this study, we characterized and conducted functional analysis of hypoxia-inducible factor HIF1α and its inhibitor HIF1αn in Nile tilapia (Oreochromis niloticus). By cloning and Sanger sequencing, we obtained the full length cDNA sequences for HIF1α (2686bp) and HIF1αn (1308bp), respectively. The CDS of HIF1α includes 15 exons encoding 768 amino acid residues and the CDS of HIF1αn contains 8 exons encoding 354 amino acid residues. The complete CDS sequences of HIF1α and HIF1αn cloned from tilapia shared very high homology with known genes from other fishes. HIF1α show differentiated expression in different tissues (brain, heart, gill, spleen, liver) and at different hypoxia exposure times (6h, 12h, 24h). HIF1αn expression level under hypoxia is generally increased (6h, 12h, 24h) and shows extremely highly upregulation in brain tissue under hypoxia. A functional determination site analysis in the protein sequences between fish and land animals identified 21 amino acid sites in HIF1α and 2 sites in HIF1αn as significantly associated sites (α = 0.05). Phylogenetic tree-based positive selection analysis suggested 22 sites in HIF1α as positively selected sites with a p-value of at least 95% for fish lineages compared to the land animals. Our study could be important for clarifying the mechanism of fish adaptation to aquatic hypoxia environment.

  18. cDNA cloning and characterization of mouse DTEF-1 and ETF, members of the TEA/ATTS family of transcription factors.

    Science.gov (United States)

    Yockey, C E; Shimizu, N

    1998-02-01

    Members of the TEA/ATTS family of transcription factors have been found in most representative eukaryotic organisms. In vertebrates, the TEA family contains at least four members, which share overlapping DNA-binding specificity and have similar transcriptional activation properties. In this article, we describe the cDNA cloning and characterization of the murine TEA proteins DTEF-1 (mDTEF-1) and ETF. Using in situ hybridization analysis of mouse embryos, we found that mDTEF-1 and ETF transcript distributions substantially overlap. ETF is expressed throughout the embryo except in the myocardium early in development, whereas late in development, it is enriched in lung and neuroectoderm. Mouse DTEF-1 is expressed at a much lower level throughout development and is substantially enriched in ectoderm and skin, as well as in the developing pituitary at midgestation. Northern blot analysis of adult mouse tissue total RNA showed that both ETF and mDTEF-1 are abundant in uterus and lung relative to other tissues. Using gel mobility shift assays and GAL4-fusion protein analysis, we demonstrated that the full coding sequences of ETF and mDTEF-1 encode M-CAT/GT-IIC-binding proteins containing activation domains.

  19. Molecular characterization and expression analysis of the first Porifera tumor necrosis factor superfamily member and of its putative receptor in the marine sponge Chondrosia reniformis.

    Science.gov (United States)

    Pozzolini, Marina; Scarfì, Sonia; Ghignone, Stefano; Mussino, Francesca; Vezzulli, Luigi; Cerrano, Carlo; Giovine, Marco

    2016-04-01

    Here we report the molecular cloning and characterization of the first Tumor Necrosis Factor homologous and of its putative receptor in the marine sponge Chondrosia reniformis: chTNF and chTNFR, respectively. The deduced chTNF amino acid sequence is a type II transmembrane protein containing the typical TNFSF domain. Phylogenetic analysis reveals that chTNF is more related to Chordata TNFs rather than to other invertebrates. chTNF and chTNFR are constitutively expressed both in the ectosome and in the choanosome of the sponge, with higher levels in the ectosome. chTNF and chTNFR mRNAs were monitored in sponge fragmorphs treated with Gram(+) or Gram(-) bacteria. chTNF was significantly upregulated in Gram(+)-treated fragmorphs as compared to controls, while chTNFR was upregulated by both treatments. Finally, the possible chTNF fibrogenic role in sponge fragmorphs was studied by TNF inhibitor treatment measuring fibrillar and non fibrillar collagen gene expression; results indicate that the cytokine is involved in sponge collagen deposition and homeostasis. Copyright © 2015 Elsevier Ltd. All rights reserved.

  20. Identification and in vitro characterization of novel nanobodies against human granulocyte colony-stimulating factor receptor to provide inhibition of G-CSF function.

    Science.gov (United States)

    Bakherad, Hamid; Gargari, Seyed Latif Mousavi; Sepehrizadeh, Zargham; Aghamollaei, Hossein; Taheri, Ramezan Ali; Torshabi, Maryam; Yazdi, Mojtaba Tabatabaei; Ebrahimizadeh, Walead; Setayesh, Neda

    2017-09-01

    It has been shown that Granulocyte colony-stimulating factor (G-CSF) has a higher expression in malignant tumors, and anti-G-CSF therapy considerably decreases tumor growth, tumor vascularization and metastasis. Thus, blocking the signaling pathway of G-CSF could be beneficial in cancer therapy. This study is aimed at designing and producing a monoclonal nanobody that could act as an antagonist of G-CSF receptor. Nanobodies are the antigen binding fragments of camelid single-chain antibodies, also known as VHH. These fragments have exceptional properties which makes them ideal for tumor imaging and therapeutic applications. We have used our previously built nanobody phage libraries to isolate specific nanobodies to the G-CSF receptor. After a series of cross-reactivity and affinity experiments, two unique nanobodies were selected for functional analysis. Proliferation assay, real-time PCR and immunofluorescence assays were used to characterize these nanobodies. Finally, VHH26 nanobody that was able to specifically bind G-CSF receptor (G-CSF-R) on the surface of NFS60 cells and efficiently block G-CSF-R downstream signaling pathway in a dose-dependent manner was selected. This nanobody could be further developed into a valuable tool in tumor therapy and it forms a basis for additional studies in preclinical animal models. Copyright © 2017. Published by Elsevier Masson SAS.

  1. Characterization of pH titration shifts for all the nonlabile proton resonances in a protein by two-dimensional NMR: The case of mouse epidermal growth factor

    International Nuclear Information System (INIS)

    Kohda, Daisuke; Sawada, Toshie; Inagaki, Fuyuhiko

    1991-01-01

    The pH titration shifts for all the nonlabile proton resonances in a 53-residue protein (mouse epidermal growth factor) were measured in the p 2 H range 1.5-9 with two-dimensional (2D) 1 H NMR. The 2D NMR pH titration experiment made it possible to determine the pK values for all the ionizable group which were titrated in the pH range 1.5-9 in the protein. The pK values of the nine ionizable groups (α-amino group, four Asp, two Glu, one His, and α-carboxyl group) were found to be near their normal values. The 2D titration experiment also provided a detailed description of the pH-dependent behavior of the proton chemical shifts and enabled us to characterize the pH-dependent changes of protein conformation. Analysis of the pH-dependent shifts of ca. 200 proton resonances offered evidence of conformational changes in slightly basic pH solution: The deprotonation of the N-terminal α-amino group induced a widespread conformational change over the β-sheet structure in the protein, while the effects of deprotonation of the His22 imidazole group were relatively localized. The authors found that the 2D NMR pH titration experiment is a powerful tool for investigating the structural and dynamic properties of proteins

  2. Isolation, Characterization, and Molecular Modeling of a Rheumatoid Factor from a Hepatitis C Virus Infected Patient with Sjögren’s Syndrome

    Directory of Open Access Journals (Sweden)

    Yu-Ching Lee

    2013-01-01

    Full Text Available We have previously isolated several IgG rheumatoid factors (RFs from patients with both rheumatoid arthritis and idiopathic thrombocytopenia purpura using phage display system. To study IgG RFs in patients with other autoimmune diseases, phage display antibody libraries from a hepatitis C virus infected patient with Sjögren’s syndrome were constructed. After panning, a specific clone RFL11 was isolated for characterization in advance. The binding activity and specificity of RFL11 to IgG Fc fragment were comparable to those of RFs previously isolated. The analysis with existed RF-Fc complex structures indicated the homology model of RFL11 is similar to IgM RF61 complex with high binding affinity of about 6×10-8 M. This effect resulted from longer complementarity-determining region (CDR combining key somatic mutations. In the RFL11-Fc interfaces, the CDR-H3 loop forms a finger-like structure extending into the bottom of Fc pocket and resulting in strong ion and cation-pi interactions. Moreover, a process of antigen-driven maturation was proven by somatically mutated VH residues on H2 and H3 CDR loops in the interfaces. Taken together, these results suggested that high affinity IgG RFs can be generated in patients with Sjögren’s syndrome and may play an important role in the pathogenesis of this autoimmune disease.

  3. GMP production and characterization of leucine zipper-tagged tumor necrosis factor-related apoptosis-inducing ligand (LZ-TRAIL) for phase I clinical trial.

    Science.gov (United States)

    Jiang, Jing; Liu, Xiaobin; Deng, Leixiu; Zhang, Peipei; Wang, Guangjun; Wang, Shifu; Liu, Honghao; Su, Yunpeng

    2014-10-05

    Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) exhibits potent antitumor activity in a wide range of cancers without deleterious side effects on normal tissues. Several TRAIL derivatives have been developed to improve its pharmacokinetics and therapeutic effects through strategies such as adding a leucine zipper to increase the circulation half-life. To obtain clinical grade LZ-TRAIL for phase I clinical trial, a single batch of 30 L bioreactor culture was performed using the Escherichia coli BL21 (DE3) strain expressing the recombinant LZ-TRAIL. A robust LZ-TRAIL production fermentation process was developed, which could be scaled up from 5L to 50 L, and had a titer of approximately 1.4 g/l. A four-step purification strategy was carried out to obtain a final product with over 95% purity and 45% yield. The final material was filter sterilized, aseptically vialed, and stored at 4°C, and comprehensively characterized using multiple assays (vialed product was sterile, purity was 95%, aggregates were production of phase I clinical trial material. These preclinical investigations warrant further clinical development of this product for cancer therapy. Copyright © 2014. Published by Elsevier B.V.

  4. Molecular characterization and expression profile of nanos in Schistosoma japonicum and its influence on the expression several mammalian stem cell factors.

    Science.gov (United States)

    Giri, Bikash Ranjan; Du, Xiaoli; Xia, Tianqi; Chen, Yongjun; Li, Hao; Cheng, Guofeng

    2017-07-01

    Pluripotent stem cells, called neoblasts, are well known for the regenerative capability and developmental plasticity in flatworms. Impressive advancement has been made in free-living flatworms, while in case of its parasitic counterpart, neoblast-like stem cells have attracted recent attention for its self-renewal and differentiation capacity. Nanos is a key conserved post-transcriptional regulator critical for the formation, development, and/or maintenance of the pluripotent germ line stem cell systems in many metazoans including schistosomes. In the present study, we report the molecular cloning and expression of nanos orthologous genes nanos in Schistosoma japonicum (Sjnanos). The cDNA of Sjnanos is 826 bp long, containing an open reading frame (ORF) for 223 amino acid long protein. qRT-PCR analysis shown that Sjnanos was differently expressed in several stages of schistosomes with relatively high level in schistosomula. Additionally, Sjnanos was expressed highly in adult females compared to adult males. Transfection of recombinant plasmid for expressing Sjnanos resulted in significant proliferation and increased expression of several stem cell factors in mammalian cells. Overall, our preliminary study provides the molecular basis to further functionally characterize Sjnanos in S. japonicum.

  5. Intact molecular characterization of cord factor (trehalose 6,6'-dimycolate) from nine species of mycobacteria by MALDI-TOF mass spectrometry.

    Science.gov (United States)

    Fujita, Yukiko; Naka, Takashi; McNeil, Michael R; Yano, Ikuya

    2005-10-01

    Cord factor (trehalose 6,6'-dimycolate, TDM) is an unique glycolipid with a trehalose and two molecules of mycolic acids in the mycobacterial cell envelope. Since TDM consists of two molecules of very long branched-chain 3-hydroxy fatty acids, the molecular mass ranges widely and in a complex manner. To characterize the molecular structure of TDM precisely and simply, an attempt was made to determine the mycolic acid subclasses of TDM and the molecular species composition of intact TDM by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry for the first time. The results showed that less than 1 microg mycolic acid methyl ester of TDM from nine representative species of mycobacteria and TDM from the same species was sufficient to obtain well-resolved mass spectra composed of pseudomolecular ions [M+Na]+. Although the mass ion distribution was extremely diverse, the molecular species of each TDM was identified clearly by constructing a molecular ion matrix consisting of the combination of two molecules of mycolic acids. The results showed a marked difference in the molecular structure of TDM among mycobacterial species and subspecies. TDM from Mycobacterium tuberculosis (H37Rv and Aoyama B) showed a distinctive mass pattern and consisted of over 60 molecular ions with alpha-, methoxy- and ketomycolate. TDM from Mycobacterium bovis BCG Tokyo 172 similarly showed over 35 molecular ions, but that from M. bovis BCG Connaught showed simpler molecular ion clusters consisting of less than 35 molecular species due to a complete lack of methoxymycolate. Mass ions due to TDM from M. bovis BCG Connaught and Mycobacterium kansasii showed a biphasic distribution, but the two major peaks of TDM from M. kansasii were shifted up two or three carbon units higher compared with M. bovis BCG Connaught. Within the rapid grower group, in TDM consisting of alpha-, keto- and wax ester mycolate from Mycobacterium phlei and Mycobacterium flavescens, the

  6. Characterization of insulin-like growth factor I and insulin receptors on cultured bovine adrenal fasciculata cells. Role of these peptides on adrenal cell function

    International Nuclear Information System (INIS)

    Penhoat, A.; Chatelain, P.G.; Jaillard, C.; Saez, J.M.

    1988-01-01

    We have characterized insulin-like growth factor I (IGF-I) and insulin receptors in cultured bovine adrenal cells by binding and cross-linking affinity experiments. At equilibrium the dissociation constant and the number of binding sites per cell for IGF-I were 1.4 +/- (SE) 0.3 x 10(-9) M and 19,200 +/- 2,100, respectively. Under reduction conditions, disuccinimidyl suberate cross-linked [ 125 I]iodo-IGF-I to one receptor complex with an Mr of 125,000. Adrenal cells also contain specific insulin receptors with an apparent dissociation constant (Kd) of 10(-9) M. Under reduction conditions [ 125 I]iodo-insulin binds to one band with an approximate Mr of 125,000. IGF-I and insulin at micromolar concentrations, but not at nanomolar concentrations, slightly stimulated DNA synthesis, but markedly potentiated the mitogenic action of fibroblast growth factor. Adrenal cells cultured in a serum-free medium containing transferrin, ascorbic acid, and insulin (5 micrograms/ml) maintained fairly constant angiotensin-II (A-II) receptor concentration per cell and increased cAMP release on response to ACTH and their steroidogenic response to both ACTH and A-II. When the cells were cultured in the same medium without insulin, the number of A-II receptors significantly decreased to 65% and the increased responsiveness was blunted. Treatment of such cells for 3 days with increasing concentrations of IGF-I (1-100 ng/ml) produced a 2- to 3-fold increase in A-II receptors and enhanced the cAMP response (3- to 4-fold) to ACTH and the steroidogenic response (4- to 6-fold) to ACTH and A-II. These effects were time and dose dependent (ED50 approximately equal to 10(-9) M). Insulin at micromolar concentrations produced an effect similar to that of IGF-I, but at nanomolar concentrations the effect was far less

  7. Characterization of bonded stationary phase performance as a function of qualitative and quantitative chromatographic factors in chaotropic chromatography with risperidone and its impurities as model substances.

    Science.gov (United States)

    Čolović, Jelena; Rmandić, Milena; Malenović, Anđelija

    2018-05-17

    Numerous stationary phases have been developed with the aim to provide desired performances during chromatographic analysis of the basic solutes in their protonated form. In this work, the procedure for the characterization of bonded stationary phase performance, when both qualitative and quantitative chromatographic factors were varied in chaotropic chromatography, was proposed. Risperidone and its three impurities were selected as model substances, while acetonitrile content in the mobile phase (20-30%), the pH of the aqueous phase (3.00-5.00), the content of chaotropic agents in the aqueous phase (10-100 mM), type of chaotropic agent (NaClO 4 , CF 3 COONa), and stationary phase type (Zorbax Eclipse XDB, Zorbax Extend) were studied as chromatographic factors. The proposed procedure implies the combination of D-optimal experimental design, indirect modeling, and polynomial-modified Gaussian model, while grid point search method was selected for the final choice of the experimental conditions which lead to the best possible stationary phase performance for basic solutes. Good agreement between experimentally obtained chromatogram and simulated chromatogram for chosen experimental conditions (25% acetonitrile, 75 mM of NaClO 4 , pH 4.00 on Zorbax Eclipse XDB column) confirmed the applicability of the proposed procedure. The additional point was selected for the verification of proposed procedure ability to distinguish changes in solutes' elution order. Simulated chromatogram for 21.5% acetonitrile, 85 mM of NaClO 4 , pH 5.00 on Zorbax Eclipse XDB column was in line with experimental data. Furthermore, the values of left and right peak half-widths obtained from indirect modeling were used in order to evaluate performances of differently modified stationary phases applying a half-width plots approach. The results from half-width plot approach as well as from the proposed procedure indicate higher efficiency and better separation performance of the stationary phase

  8. Characterization of common carp (Cyprinus carpio L.) interferon regulatory factor 5 (IRF5) and its expression in response to viral and bacterial challenges.

    Science.gov (United States)

    Zhu, Yaoyao; Qi, Chenchen; Shan, Shijuan; Zhang, Fumiao; Li, Hua; An, Liguo; Yang, Guiwen

    2016-06-27

    Common carp (Cyprinus carpio L.), one of the most economically valuable commercial farming fish species in China, is often infected by a variety of viruses. As the first line of defence against microbial pathogens, the innate immune system plays a crucial role in teleost fish, which are lower vertebrates. Interferon (IFN) regulatory factor 5 (IRF5) is a key molecule in antiviral immunity that regulating the expression of IFN and other pro-inflammatory cytokines. It is necessary to gain more insight into the common carp IFN system and the function of fish IRF5 in the antiviral and antibacterial response. In the present study, we characterized the cDNA and genomic sequence of the IRF5 gene in common carp, and analysed tissue distribution and expression profile of this gene in response to polyinosinic:polycytidylic acid (poly I:C) and lipopolysaccharides (LPS) treatment. The common carp IRF5 (ccIRF5) gene is 5790 bp in length and is composed of 9 exons and 8 introns. The open reading frame (ORF) of ccIRF5 is 1554 bp, and encodes 517 amino acid protein. The putative ccIRF5 protein shares identity (65.4-90.0 %) with other fish IRF5s and contains a DNA binding domain (DBD), a middle region (MR), an IRF-associated domain (IAD), a virus activated domain (VAD) and two nuclear localization signals (NLSs) similar to those found in vertebrate IRF5. Phylogenetic analysis clustered ccIRF5 into the IRF5 subfamily with other vertebrate IRF5 and IRF6 genes. Real-time PCR analysis revealed that ccIRF5 mRNA was expressed in all examined tissues of healthy carps, with high levels observed in the gills and the brain. After poly I:C challenge, expression levels of ccIRF5, tumour-necrosis factor α (ccTNFα) and two IFN stimulated genes [ISGs (ccISG5 and ccPKR)] were up-regulated in seven immune-related tissues (liver, spleen, head kidney, foregut, hindgut, skin and gills). Furthermore, all four genes were up-regulated in vitro upon poly I:C and LPS challenges. Our findings suggest

  9. Characterization of buffy coat-derived granulocytes for clinical use: a comparison with granulocyte colony-stimulating factor/dexamethasone-pretreated donor-derived products.

    Science.gov (United States)

    van de Geer, A; Gazendam, R P; Tool, A T J; van Hamme, J L; de Korte, D; van den Berg, T K; Zeerleder, S S; Kuijpers, T W

    2017-02-01

    Buffy coat-derived granulocytes have been described as an alternative to the apheresis product from donors pretreated with dexamethasone and granulocyte colony-stimulating factor (G-CSF). The latter is - dependent on the local and national settings - obtained following a demanding and time-consuming procedure, which is undesirable in critically ill septic patients. In contrast, buffy coat-derived products have a large volume and are often heavily contaminated with red cells and platelets. We developed a new pooled buffy coat-derived product with high purity and small volume, and performed a comprehensive functional characterization of these granulocytes. We pooled ten buffy coats following the production of platelet concentrates. Saline 0·9% was added to decrease the viscosity and the product was split into plasma, red cells and a 'super' buffy coat. Functional data of the granulocytes were compared to those obtained with granulocytes from healthy controls and G-CSF/dexamethasone-pretreated donors. Buffy coat-derived granulocytes showed adhesion, chemotaxis, reactive oxygen species production, degranulation, NETosis and in vitro killing of Staphylococcus aureus, Escherichia coli and Aspergillus species comparable to control and G-CSF/dexamethasone-derived granulocytes. Candida killing was superior compared to G-CSF/dexamethasone-derived granulocytes. Immunophenotyping was normal; especially no signs of activation in the buffy coat-derived granulocytes were seen. Viability was reduced. Buffy coats are readily available in the regular blood production process and would take away the concerns around the apheresis product. The product described appears a promising alternative for transfusion purposes. © 2017 International Society of Blood Transfusion.

  10. Increased levels of NOTCH1, NF-kappaB, and other interconnected transcription factors characterize primitive sets of hematopoietic stem cells.

    Science.gov (United States)

    Panepucci, Rodrigo Alexandre; Oliveira, Lucila Habib B; Zanette, Dalila Luciola; Viu Carrara, Rita de Cassia; Araujo, Amélia Goes; Orellana, Maristela Delgado; Bonini de Palma, Patrícia Vianna; Menezes, Camila C B O; Covas, Dimas Tadeu; Zago, Marco Antonio

    2010-03-01

    As previously shown, higher levels of NOTCH1 and increased NF-kappaB signaling is a distinctive feature of the more primitive umbilical cord blood (UCB) CD34+ hematopoietic stem cells (HSCs), as compared to bone marrow (BM). Differences between BM and UCB cell composition also account for this finding. The CD133 marker defines a more primitive cell subset among CD34+ HSC with a proposed hemangioblast potential. To further evaluate the molecular basis related to the more primitive characteristics of UCB and CD133+ HSC, immunomagnetically purified human CD34+ and CD133+ cells from BM and UCB were used on gene expression microarrays studies. UCB CD34+ cells contained a significantly higher proportion of CD133+ cells than BM (70% and 40%, respectively). Cluster analysis showed that BM CD133+ cells grouped with the UCB cells (CD133+ and CD34+) rather than to BM CD34+ cells. Compared with CD34+ cells, CD133+ had a higher expression of many transcription factors (TFs). Promoter analysis on all these TF genes revealed a significantly higher frequency (than expected by chance) of NF-kappaB-binding sites (BS), including potentially novel NF-kappaB targets such as RUNX1, GATA3, and USF1. Selected transcripts of TF related to primitive hematopoiesis and self-renewal, such as RUNX1, GATA3, USF1, TAL1, HOXA9, HOXB4, NOTCH1, RELB, and NFKB2 were evaluated by real-time PCR and were all significantly positively correlated. Taken together, our data indicate the existence of an interconnected transcriptional network characterized by higher levels of NOTCH1, NF-kappaB, and other important TFs on more primitive HSC sets.

  11. Genome-wide identification and characterization of cacao WRKY transcription factors and analysis of their expression in response to witches' broom disease.

    Science.gov (United States)

    Silva Monteiro de Almeida, Dayanne; Oliveira Jordão do Amaral, Daniel; Del-Bem, Luiz-Eduardo; Bronze Dos Santos, Emily; Santana Silva, Raner José; Peres Gramacho, Karina; Vincentz, Michel; Micheli, Fabienne

    2017-01-01

    Transcriptional regulation, led by transcription factors (TFs) such as those of the WRKY family, is a mechanism used by the organism to enhance or repress gene expression in response to stimuli. Here, we report on the genome-wide analysis of the Theobroma cacao WRKY TF family and also investigate the expression of WRKY genes in cacao infected by the fungus Moniliophthora perniciosa. In the cacao genome, 61 non-redundant WRKY sequences were found and classified in three groups (I to III) according to the WRKY and zinc-finger motif types. The 61 putative WRKY sequences were distributed on the 10 cacao chromosomes and 24 of them came from duplication events. The sequences were phylogenetically organized according to the general WRKY groups. The phylogenetic analysis revealed that subgroups IIa and IIb are sister groups and share a common ancestor, as well as subgroups IId and IIe. The most divergent groups according to the plant origin were IIc and III. According to the phylogenetic analysis, 7 TcWRKY genes were selected and analyzed by RT-qPCR in susceptible and resistant cacao plants infected (or not) with M. perniciosa. Some TcWRKY genes presented interesting responses to M. perniciosa such as Tc01_p014750/Tc06_p013130/AtWRKY28, Tc09_p001530/Tc06_p004420/AtWRKY40, Tc04_p016130/AtWRKY54 and Tc10_p016570/ AtWRKY70. Our results can help to select appropriate candidate genes for further characterization in cacao or in other Theobroma species.

  12. Characterization of the human nasal embryonic LHRH factor gene, NELF, and a mutation screening among 65 patients with idiopathic hypogonadotropic hypogonadism (IHH).

    Science.gov (United States)

    Miura, Kiyonori; Acierno, James S; Seminara, Stephanie B

    2004-01-01

    As the mouse nasal embryonic LHRH factor gene (Nelf) encodes a guidance molecule for the migration of the olfactory axon and gonadotropin-releasing hormone neurons, its human homolog, NELF, is a candidate gene for Kallmann syndrome, a disease of idiopathic hypogonadotropic hypogonadism (IHH) with anosmia or hyposmia. We report here characterization of NELF and results of mutation analysis in 65 IHH patients. Assembling EST clones, RACE, and sequencing showed that NELF mapped to 9q34.3 is composed of 16 exons and 15 introns with a 1,590-bp ORF encoding 530 amino acids. RT-PCR on a fetal brain cDNA library revealed five alternatively spliced variants. Among them, NELF-v1 has 93-94% identity at the amino acid level to mouse/rat Nelf, and four other transcripts are also highly conserved among the three species. A 3.0-kb transcript is expressed most highly in the adult and fetal brain, testis, and kidney, indicating that NELF plays a role in the function of these tissues. Mutation screening detected in a patient with IHH one novel heterozygous missense mutation (1438A>G, T480A) at the donor-splice site in exon 15 of NELF. As this mutation was not found in 100 normal control individuals, T480A may be associated with IHH. Four other novel SNPs (102C > T and 1029C > T within the coding region, and two IVS14+47C > T and IVS15+41G > A) were also identified in NELF.

  13. Genome-wide identification and characterization of cacao WRKY transcription factors and analysis of their expression in response to witches' broom disease

    Science.gov (United States)

    Silva Monteiro de Almeida, Dayanne; Oliveira Jordão do Amaral, Daniel; Del-Bem, Luiz-Eduardo; Bronze dos Santos, Emily; Santana Silva, Raner José; Peres Gramacho, Karina; Vincentz, Michel

    2017-01-01

    Transcriptional regulation, led by transcription factors (TFs) such as those of the WRKY family, is a mechanism used by the organism to enhance or repress gene expression in response to stimuli. Here, we report on the genome-wide analysis of the Theobroma cacao WRKY TF family and also investigate the expression of WRKY genes in cacao infected by the fungus Moniliophthora perniciosa. In the cacao genome, 61 non-redundant WRKY sequences were found and classified in three groups (I to III) according to the WRKY and zinc-finger motif types. The 61 putative WRKY sequences were distributed on the 10 cacao chromosomes and 24 of them came from duplication events. The sequences were phylogenetically organized according to the general WRKY groups. The phylogenetic analysis revealed that subgroups IIa and IIb are sister groups and share a common ancestor, as well as subgroups IId and IIe. The most divergent groups according to the plant origin were IIc and III. According to the phylogenetic analysis, 7 TcWRKY genes were selected and analyzed by RT-qPCR in susceptible and resistant cacao plants infected (or not) with M. perniciosa. Some TcWRKY genes presented interesting responses to M. perniciosa such as Tc01_p014750/Tc06_p013130/AtWRKY28, Tc09_p001530/Tc06_p004420/AtWRKY40, Tc04_p016130/AtWRKY54 and Tc10_p016570/ AtWRKY70. Our results can help to select appropriate candidate genes for further characterization in cacao or in other Theobroma species. PMID:29084273

  14. Characterizing chromophoric dissolved organic matter in Lake Tianmuhu and its catchment basin using excitation-emission matrix fluorescence and parallel factor analysis.

    Science.gov (United States)

    Zhang, Yunlin; Yin, Yan; Feng, Longqing; Zhu, Guangwei; Shi, Zhiqiang; Liu, Xiaohan; Zhang, Yuanzhi

    2011-10-15

    Chromophoric dissolved organic matter (CDOM) is an important optically active substance that transports nutrients, heavy metals, and other pollutants from terrestrial to aquatic systems and is used as a measure of water quality. To investigate how the source and composition of CDOM changes in both space and time, we used chemical, spectroscopic, and fluorescence analyses to characterize CDOM in Lake Tianmuhu (a drinking water source) and its catchment in China. Parallel factor analysis (PARAFAC) identified three individual fluorophore moieties that were attributed to humic-like and protein-like materials in 224 water samples collected between December 2008 and September 2009. The upstream rivers contained significantly higher concentrations of CDOM than did the lake water (a(350) of 4.27±2.51 and 2.32±0.59 m(-1), respectively), indicating that the rivers carried a substantial load of organic matter to the lake. Of the three main rivers that flow into Lake Tianmuhu, the Pingqiao River brought in the most CDOM from the catchment to the lake. CDOM absorption and the microbial and terrestrial humic-like components, but not the protein-like component, were significantly higher in the wet season than in other seasons, indicating that the frequency of rainfall and runoff could significantly impact the quantity and quality of CDOM collected from the catchment. The different relationships between the maximum fluorescence intensities of the three PARAFAC components, CDOM absorption, and chemical oxygen demand (COD) concentration in riverine and lake water indicated the difference in the composition of CDOM between Lake Tianmuhu and the rivers that feed it. This study demonstrates the utility of combining excitation-emission matrix fluorescence and PARAFAC to study CDOM dynamics in inland waters. Copyright © 2011 Elsevier Ltd. All rights reserved.

  15. Phenotypic characterization of a novel virulence-factor deletion strain of Burkholderia mallei that provides partial protection against inhalational glanders in mice

    Directory of Open Access Journals (Sweden)

    Joel A. Bozue

    2016-02-01

    Full Text Available Burkholderia mallei (Bm is a highly infectious intracellular pathogen classified as a category B biological agent by the Centers for Disease Control and Prevention. After respiratory exposure, Bm establishes itself within host macrophages before spreading into major organ systems, which can lead to chronic infection, sepsis, and death. Previously, we combined computational prediction of host-pathogen interactions with yeast two-hybrid experiments and identified novel virulence factor genes in Bm, including BMAA0553, BMAA0728 (tssN, and BMAA1865. In the present study, we used recombinant allelic exchange to construct deletion mutants of BMAA0553 and tssN (ΔBMAA0553 and ΔTssN, respectively and showed that both deletions completely abrogated virulence at doses of >100 times the LD50 of the wild-type Bm strain. Analysis of ΔBMAA0553- and ΔTssN-infected mice showed starkly reduced bacterial dissemination relative to wild-type Bm, and subsequent in vitro experiments characterized pathogenic phenotypes with respect to intracellular growth, macrophage uptake and phagosomal escape, actin-based motility, and multinucleated giant cell formation. Based on observed in vitro and in vivo phenotypes, we explored the use of ΔTssN as a candidate live-attenuated vaccine. Mice immunized with aerosolized ΔTssN showed a 21-day survival rate of 67% after a high-dose aerosol challenge with the wild-type Bm ATCC 23344 strain, compared to a 0% survival rate for unvaccinated mice. However, analysis of histopathology and bacterial burden showed that while the surviving vaccinated mice were protected from acute infection, Bm was still able to establish a chronic infection. Vaccinated mice showed a modest IgG response, suggesting a limited potential of ΔTssN as a vaccine candidate, but also showed prolonged elevation of pro-inflammatory cytokines, underscoring the role of cellular and innate immunity in mitigating acute infection in inhalational glanders.

  16. Characterization of the snowy cotyledon 1 mutant of Arabidopsis thaliana: the impact of chloroplast elongation factor G on chloroplast development and plant vitality.

    Science.gov (United States)

    Albrecht, Verónica; Ingenfeld, Anke; Apel, Klaus

    2006-03-01

    During seedling development chloroplast formation marks the transition from heterotrophic to autotrophic growth. The development and activity of chloroplasts may differ in cotyledons that initially serve as a storage organ and true leaves whose primary function is photosynthesis. A genetic screen was used for the identification of genes that affect selectively chloroplast function in cotyledons of Arabidopsis thaliana. Several mutants exhibiting pale cotyledons and green true leaves were isolated and dubbed snowy cotyledon (sco). One of the mutants, sco1, was characterized in more detail. The mutated gene was identified using map-based cloning. The mutant contains a point mutation in a gene encoding the chloroplast elongation factor G, leading to an amino acid exchange within the predicted 70S ribosome-binding domain. The mutation results in a delay in the onset of germination. At this early developmental stage embryos still contain undifferentiated proplastids, whose proper function seems necessary for seed germination. In light-grown sco1 seedlings the greening of cotyledons is severely impaired, whereas the following true leaves develop normally as in wild-type plants. Despite this apparent similarity of chloroplast development in true leaves of mutant and wild-type plants various aspects of mature plant development are also affected by the sco1 mutation such as the onset of flowering, the growth rate, and seed production. The onset of senescence in the mutant and the wild-type plants occurs, however, at the same time, suggesting that in the mutant this particular developmental step does not seem to suffer from reduced protein translation efficiency in chloroplasts.

  17. Structural characterization of a novel full-length transcript promoter from Horseradish Latent Virus (HRLV) and its transcriptional regulation by multiple stress responsive transcription factors.

    Science.gov (United States)

    Khan, Ahamed; Shrestha, Ankita; Bhuyan, Kashyap; Maiti, Indu B; Dey, Nrisingha

    2018-01-01

    The promoter fragment described in this study can be employed for strong transgene expression under both biotic and abiotic stress conditions. Plant-infecting Caulimoviruses have evolved multiple regulatory mechanisms to address various environmental stimuli during the course of evolution. One such mechanism involves the retention of discrete stress responsive cis-elements which are required for their survival and host-specificity. Here we describe the characterization of a novel Caulimoviral promoter isolated from Horseradish Latent Virus (HRLV) and its regulation by multiple stress responsive Transcription factors (TFs) namely DREB1, AREB1 and TGA1a. The activity of full length transcript (Flt-) promoter from HRLV (- 677 to + 283) was investigated in both transient and transgenic assays where we identified H12 (- 427 to + 73) as the highest expressing fragment having ~ 2.5-fold stronger activity than the CaMV35S promoter. The H12 promoter was highly active and near-constitutive in the vegetative and reproductive parts of both Tobacco and Arabidopsis transgenic plants. Interestingly, H12 contains a distinct cluster of cis-elements like dehydration-responsive element (DRE-core; GCCGAC), an ABA-responsive element (ABRE; ACGTGTC) and as-1 element (TGACG) which are known to be induced by cold, drought and pathogen/SA respectively. The specific binding of DREB1, AREB1 and TGA1a to DRE, ABRE and as-1 elements respectively were confirmed by the gel-binding assays using H12 promoter-specific probes. Detailed mutational analysis of the H12 promoter suggested that the presence of DRE-core and as-1 element was indispensable for its activity which was further confirmed by the transactivation assays. Our studies imply that H12 could be a valuable genetic tool for regulated transgene expression under diverse environmental conditions.

  18. Synthesis, purification, and characterization of an Arg152 → Glu site-directed mutant of recombinant human blood clotting factor VII

    International Nuclear Information System (INIS)

    Wildgoose, P.; Kisiel, W.; Berkner, K.L.

    1990-01-01

    Coagulation factor VII circulates in blood as a single-chain zymogen of a serine protease and is converted to its activated two-chain form, factor VIIa, by cleavage of an internal peptide bond located at Arg 152 -Ile 153 . Previous studies using serine protease active-site inhibitors suggest that zymogen factor VII may possess sufficient proteolytic activity to initiate the extrinsic pathway of blood coagulation. In order to assess the putative intrinsic proteolytic activity of single-chain factor VII, the authors have constructed a site-specific mutant of recombinant human factor VII in which arginine-152 has been replaced with a glutamic acid residue. Mutant factor VII was purified in a single step from culture supernatants of baby hamster kidney cells transfected with a plasmid containing the sequence for Arg 152 → Glu factor VII using a calcium-dependent, murine anti-factor VII monoclonal antibody column. The clotting activity of mutant factor VII was completely inhibited following incubation with dansyl-Glu-Gly-Arg chloromethyl ketone, suggesting that the apparent clotting activity of mutant factor VII was due to a contaminating serine protease. Immunoblots of mutant factor VII with human factor IXa revealed no cleavage, whereas incubation of mutant factor VII with human factor Xa resulted in cleavage of mutant factor VII and the formation of a lower molecular weight degradation product migrating at M r ∼40 000. The results are consistent with the proposal that zymogen factor VII possesses no intrinsic proteolytic activity toward factor X or factor IX

  19. Synthesis, purification, and characterization of an Arg sub 152 yields Glu site-directed mutant of recombinant human blood clotting factor VII

    Energy Technology Data Exchange (ETDEWEB)

    Wildgoose, P.; Kisiel, W. (Univ. of New Mexico, Albuquerque (USA)); Berkner, K.L. (ZymoGenetics, Inc., Seattle, WA (USA))

    1990-04-03

    Coagulation factor VII circulates in blood as a single-chain zymogen of a serine protease and is converted to its activated two-chain form, factor VIIa, by cleavage of an internal peptide bond located at Arg{sub 152}-Ile{sub 153}. Previous studies using serine protease active-site inhibitors suggest that zymogen factor VII may possess sufficient proteolytic activity to initiate the extrinsic pathway of blood coagulation. In order to assess the putative intrinsic proteolytic activity of single-chain factor VII, the authors have constructed a site-specific mutant of recombinant human factor VII in which arginine-152 has been replaced with a glutamic acid residue. Mutant factor VII was purified in a single step from culture supernatants of baby hamster kidney cells transfected with a plasmid containing the sequence for Arg{sub 152} {yields} Glu factor VII using a calcium-dependent, murine anti-factor VII monoclonal antibody column. The clotting activity of mutant factor VII was completely inhibited following incubation with dansyl-Glu-Gly-Arg chloromethyl ketone, suggesting that the apparent clotting activity of mutant factor VII was due to a contaminating serine protease. Immunoblots of mutant factor VII with human factor IXa revealed no cleavage, whereas incubation of mutant factor VII with human factor Xa resulted in cleavage of mutant factor VII and the formation of a lower molecular weight degradation product migrating at M{sup r}{approx}40 000. The results are consistent with the proposal that zymogen factor VII possesses no intrinsic proteolytic activity toward factor X or factor IX.

  20. Characterization of the factor VIII defect in 147 patients with sporadic hemophilia A: Family studies indicate a mutation type-dependent sex ratio of mutation frequencies

    Energy Technology Data Exchange (ETDEWEB)

    Becker, J.; Schmidt, W.; Olek, K. [Univ. of Bonn (Germany)] [and others

    1996-04-01

    The clinical manifestation of hemophilia A is caused by a wide range of different mutations. In this study the factor VIII genes of 147 severe hemophilia A patients-all exclusively from sporadic families-were screened for mutations by use of the complete panel of modern DNA techniques. The pathogenous defect could be characterized in 126 patients (85.7%). Fifty-five patients (37.4%) showed a F8A-gene inversion, 47 (32.0%) a point mutation, 14 (9.5%) a small deletion, 8 (5.4%) a large deletion, and 2 (1.4%) a small insertion. Further, four (2.7%) mutations were localized but could not be sequenced yet. No mutation could be identified in 17 patients (11.6%). Sixteen (10.9%) of the P identified mutations occurred in the B domain. Four of these were located in an adenosine nucleotide stretch at codon 1192, indicating a mutation hotspot. Somatic mosaicisms were detected in 3 (3.9%) of 76 patients` mothers, comprising 3 of 16 de novo mutations in the patients` mothers. Investigation of family relatives allowed detection of a de novo mutation in 16 of 76 two-generation and 28 of 34 three-generation families. On the basis of these data, the male:female ratio of mutation frequencies (k) was estimated as k = 3.6. By use of the quotients of mutation origin in maternal grandfather to patient`s mother or to maternal grandmother, k was directly estimated as k = 15 and k = 7.5, respectively. Considering each mutation type separately, we revealed a mutation type-specific sex ratio of mutation frequencies. Point mutations showed a 5-to-10-fold-higher and inversions a >10-fold- higher mutation rate in male germ cells, whereas deletions showed a >5-fold-higher mutation rate in female germ cells. Consequently, and in accordance with the data of other diseases like Duchenne muscular dystrophy, our results indicate that at least for X-chromosomal disorders the male:female mutation rate of a disease is determined by its proportion of the different mutation types. 68 refs., 1 fig., 5 tabs.

  1. Greater-than-Class C low-level radioactive waste characterization. Appendix E-4: Packaging factors for greater-than-Class C low-level radioactive waste

    International Nuclear Information System (INIS)

    Quinn, G.; Grant, P.; Winberg, M.; Williams, K.

    1994-09-01

    This report estimates packaging factors for several waste types that are potential greater-than-Class C (GTCC) low-level radioactive waste (LLW). The packaging factor is defined as the volume of a GTCC LLW disposal container divided by the as-generated or ''unpackaged'' volume of the waste loaded into the disposal container. Packaging factors reflect any processes that reduce or increase an original unpackaged volume of GTCC LLW, the volume inside a waste container not occupied by the waste, and the volume of the waste container itself. Three values are developed that represent (a) the base case or most likely value for a packaging factor, (b) a high case packaging factor that corresponds to the largest anticipated disposal volume of waste, and (c) a low case packaging factor for the smallest volume expected. GTCC LLW is placed in three categories for evaluation in this report: activated metals, sealed sources, and all other waste

  2. Change of Patient-Reported Aesthetic Outcome Over Time and Identification of Factors Characterizing Poor Aesthetic Outcome After Breast-Conserving Therapy: Long-Term Results of a Prospective Cohort Study.

    Science.gov (United States)

    Hennigs, André; Biehl, Hannah; Rauch, Geraldine; Golatta, Michael; Tabatabai, Patrik; Domschke, Christoph; Schott, Sarah; Wallwiener, Markus; Schütz, Florian; Sohn, Christof; Heil, Jörg

    2016-05-01

    We analyzed the change of aesthetic outcome (AO) over time and explored factors characterizing poor AO after breast-conserving surgery (BCS). This prospective single-center cohort study included 849 patients preoperatively planned for BCS between September 2007 and December 2011. Long-term follow-up was made once in 2013. AO was measured by the Aesthetic Status (AS) of the Breast Cancer Treatment Outcome Scale questionnaire. Clinical, surgical, and pathologic variables were evaluated to identify predictors of poor AO. We applied single factor variance analyses and univariable logistic regression analyses for outcome analysis. The long-term follow-up rate in 2013 was 73 % (621 nonrecurrent with final BCS). A poor or fair AO was reported in 30 (4.8 %) and 98 (15.8 %) of these 621 patients, respectively. Single factor variance analysis showed a negative impact of higher specimen weight on AO (p procedures.

  3. Purification and characterization of factor VIII 1,689-Cys: a nonfunctional cofactor occurring in a patient with severe hemophilia A.

    Science.gov (United States)

    O'Brien, D P; Tuddenham, E G

    1989-06-01

    We have purified the factor VIII from a CRM+ Hemophilia A plasma (90 U/dL VIII:Ag but 0 U/dL VIII:C) and analyzed the protein before and after thrombin activation by Western blotting with monoclonal antibodies (MoAbs). Normal or patient citrated plasma was ultracentrifuged, cryo-ethanol-precipitated and chromatographed on Sepharose 6B. The void volume fractions were reduced and subjected to ion exchange chromatography yielding material of specific activity approximately 1,000 U/mg protein (VIII:C or VIII:Ag). Factor VIII purified in this way from normal plasma is fully activatable by thrombin with proteolytic fragmentation as previously described by F. Rotblat et al (Biochemistry 24: 4294, 1985). Factor VIII 1,689-Cys has the normal distribution of factor VIII light and heavy chains prior to thrombin activation. After exposure to thrombin the heavy chain polypeptides were fully proteolysed but the light chain was totally resistant to cleavage. This is consistent with the demonstration in the patient's leucocyte DNA of a C to T transition in codon 1,689 converting Arg to Cys at the light chain thrombin cleavage site as previously described by J. Gitschier et al (Blood 72:1022, 1988). Uncleaved light chain of Factor VIII 1,689-Cys is not released from von Willebrand factor (vWF) by thrombin, but this is not the sole cause of the functional defect since the protein purified free of vWF has no coagulant activity. We conclude that the functional defect in factor VIII 1,689-Cys is a consequence of failure to release the acidic peptide from the light chain upon thrombin activation.

  4. Characterization of a New Pink-Fruited Tomato Mutant Results in the Identification of a Null Allele of the SlMYB12 Transcription Factor.

    Science.gov (United States)

    Fernandez-Moreno, Josefina-Patricia; Tzfadia, Oren; Forment, Javier; Presa, Silvia; Rogachev, Ilana; Meir, Sagit; Orzaez, Diego; Aharoni, Aspah; Granell, Antonio

    2016-07-01

    The identification and characterization of new tomato (Solanum lycopersicum) mutants affected in fruit pigmentation and nutritional content can provide valuable insights into the underlying biology, as well as a source of new alleles for breeding programs. To date, all characterized pink-pigmented tomato fruit mutants appear to result from low SlMYB12 transcript levels in the fruit skin. Two new mutant lines displaying a pink fruit phenotype (pf1 and pf2) were characterized in this study. In the pf mutants, SlMYB12 transcripts accumulated to wild-type levels but exhibited the same truncation, which resulted in the absence of the essential MYB activation domain coding region. Allelism and complementation tests revealed that both pf mutants were allelic to the y locus and showed the same recessive null allele in homozygosis: Δy A set of molecular and metabolic effects, reminiscent of those observed in the Arabidopsis (Arabidopsis thaliana) myb11 myb12 myb111 triple mutant, were found in the tomato Δy mutants. To our knowledge, these have not been described previously, and our data support the idea of their being null mutants, in contrast to previously described transcriptional hypomorphic pink fruit lines. We detected a reduction in the expression of several flavonol glycosides and some associated glycosyl transferases. Transcriptome analysis further revealed that the effects of the pf mutations extended beyond the flavonoid pathway into the interface between primary and secondary metabolism. Finally, screening for Myb-binding sites in the candidate gene promoter sequences revealed that 141 of the 152 co-down-regulated genes may be direct targets of SlMYB12 regulation. © 2016 American Society of Plant Biologists. All Rights Reserved.

  5. Functional characterization of an alpha-factor-like Sordaria macrospora peptide pheromone and analysis of its interaction with its cognate receptor in Saccharomyces cerevisiae.

    Science.gov (United States)

    Mayrhofer, Severine; Pöggeler, Stefanie

    2005-04-01

    The homothallic filamentous ascomycete Sordaria macrospora possesses genes which are thought to encode two pheromone precursors and two seven-transmembrane pheromone receptors. The pheromone precursor genes are termed ppg1 and ppg2. The putative products derived from the gene sequence show structural similarity to the alpha-factor precursors and a-factor precursors of the yeast Saccharomyces cerevisiae. Likewise, sequence similarity has been found between the putative products of the pheromone receptor genes pre2 and pre1 and the S. cerevisiae Ste2p alpha-factor receptor and Ste3p a-factor receptor, respectively. To investigate whether the alpha-factor-like pheromone-receptor pair of S. macrospora is functional, a heterologous yeast assay was used. Our results show that the S. macrospora alpha-factor-like pheromone precursor PPG1 is processed into an active pheromone by yeast MATalpha cells. The S. macrospora PRE2 protein was demonstrated to be a peptide pheromone receptor. In yeast MATa cells lacking the endogenous Ste2p receptor, the S. macrospora PRE2 receptor facilitated all aspects of the pheromone response. Using a synthetic peptide, we can now predict the sequence of one active form of the S. macrospora peptide pheromone. We proved that S. macrospora wild-type strains secrete an active pheromone into the culture medium and that disruption of the ppg1 gene in S. macrospora prevents pheromone production. However, loss of the ppg1 gene does not affect vegetative growth or fertility. Finally, we established the yeast assay as an easy and useful system for analyzing pheromone production in developmental mutants of S. macrospora.

  6. Cloning and characterization of the .I.str ./I.operon and elongation factor Tu expression in .I.Bacillus stearothermophilus./I..

    Czech Academy of Sciences Publication Activity Database

    Krásný, Libor; Vacík, Tomáš; Fučík, Vladimír; Jonák, Jiří

    2000-01-01

    Roč. 182, č. 21 (2000), s. 6114-6122 ISSN 0021-9193 R&D Projects: GA ČR GA204/98/0863 Grant - others:HHMI(US) 75195-540305 Institutional research plan: CEZ:AV0Z5052915 Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.506, year: 2000

  7. Characterization of O-glycosylated precursors of insulin-like growth factor II by matrix-assisted laser desorption/ionization mass spectrometry

    NARCIS (Netherlands)

    Jespersen, S.; Koedam, J.A.; Hoogerbrugge, C.M.; Tjaden, U.R.; Greef, J. van der; Brande, J.L. van den

    1996-01-01

    High molecular weight precursors of insulin-like growth factor II (IGF-II) were isolated from Cohn fraction IV of human plasma by ultrafiltration, affinity chromatography and reversed-phase high-performance liquid chromatography. Molecular weight determination by matrix-assisted laser

  8. Characterizing environmental risk factors for West Nile virus in Quebec, Canada, using clinical data in humans and serology in pet dogs.

    Science.gov (United States)

    Rocheleau, J P; Michel, P; Lindsay, L R; Drebot, M; Dibernardo, A; Ogden, N H; Fortin, A; Arsenault, J

    2017-10-01

    The identification of specific environments sustaining emerging arbovirus amplification and transmission to humans is a key component of public health intervention planning. This study aimed at identifying environmental factors associated with West Nile virus (WNV) infections in southern Quebec, Canada, by modelling and jointly interpreting aggregated clinical data in humans and serological data in pet dogs. Environmental risk factors were estimated in humans by negative binomial regression based on a dataset of 191 human WNV clinical cases reported in the study area between 2011 and 2014. Risk factors for infection in dogs were evaluated by logistic and negative binomial models based on a dataset including WNV serological results from 1442 dogs sampled from the same geographical area in 2013. Forested lands were identified as low-risk environments in humans. Agricultural lands represented higher risk environments for dogs. Environments identified as impacting risk in the current study were somewhat different from those identified in other studies conducted in north-eastern USA, which reported higher risk in suburban environments. In the context of the current study, combining human and animal data allowed a more comprehensive and possibly a more accurate view of environmental WNV risk factors to be obtained than by studying aggregated human data alone.

  9. Gene structure, cDNA characterization and RNAi-based functional analysis of a myeloid differentiation factor 88 homolog in Tenebrio molitor larvae exposed to Staphylococcus aureus infection.

    Science.gov (United States)

    Patnaik, Bharat Bhusan; Patnaik, Hongray Howrelia; Seo, Gi Won; Jo, Yong Hun; Lee, Yong Seok; Lee, Bok Luel; Han, Yeon Soo

    2014-10-01

    Myeloid differentiation factor 88 (MyD88), an intracellular adaptor protein involved in Toll/Toll-like receptor (TLR) signal processing, triggers activation of nuclear factor-kappaB (NF-κB) transcription factors. In the present study, we analyzed the gene structure and biological function of MyD88 in a coleopteran insect, Tenebrio molitor (TmMyD88). The TmMyD88 gene was 1380 bp in length and consisted of five exons and four introns. The 5'-flanking sequence revealed several putative transcription factor binding sites, such as STAT-4, AP-1, cJun, cfos, NF-1 and many heat shock factor binding elements. The cDNA contained a typical death domain, a conservative Toll-like interleukin-1 receptor (TIR) domain, and a C-terminal extension (CTE). The TmMyD88 TIR domain showed three significantly conserved motifs for interacting with the TIR domain of TLRs. TmMyD88 was grouped within the invertebrate cluster of the phylogenetic tree and shared 75% sequence identity with the TIR domain of Tribolium castaneum MyD88. Homology modeling of the TmMyD88 TIR domain revealed five parallel β-strands surrounded by five α-helices that adopted loop conformations to function as an adaptor. TmMyD88 expression was upregulated 7.3- and 4.79-fold after 12 and 6h, respectively, of challenge with Staphylococcus aureus and fungal β-1,3 glucan. Silencing of the TmMyD88 transcript by RNA interference led to reduced resistance of the host to infection by S. aureus. These results indicate that TmMyD88 is required for survival against Staphylococcus infection. Copyright © 2014 Elsevier Ltd. All rights reserved.

  10. Seroprevalence of Brucella spp. in Cattle, Molecular Characterization in Milk, and the Analysis of Associated Risk Factors with Seroprevalence in Humans, Egypt.

    Science.gov (United States)

    El-Diasty, Mohamed M; Ahmed, Heba A; Sayour, Ashraf E; El Hofy, Fatma I; Tahoun, Asmaa B M B; Shafik, Saleh M

    2016-12-01

    The objective of the present study was to estimate the seroprevalence of Brucella spp. in humans and cattle at Sharkia Governorate, Egypt. In addition, identification of Brucella spp. in milk samples by PCR and culture with the evaluation of the risk factors associated with Brucella spp. seroprevalence in humans were carried out. Overall, the seroprevalence of Brucella antibodies in the examined cattle was 23.8%, while in human participants it was 21%. The examination of 205 milk samples using PCR revealed that 6.3% were positive for B. abortus biovar 1 and the results were confirmed by culture methods. Multivariate logistic regression revealed that consumption of unpasteurized dairy products, occupational contact with animals, and knowledge about the disease are risk factors associated with infection in humans. This study documented the endemic status of brucellosis in Egypt. Hygienic measures and increased awareness about the disease are recommended to minimize the spread of infection from animals to humans.

  11. ERP, a new member of the ets transcription factor/oncoprotein family: cloning, characterization, and differential expression during B-lymphocyte development.

    Science.gov (United States)

    Lopez, M; Oettgen, P; Akbarali, Y; Dendorfer, U; Libermann, T A

    1994-05-01

    The ets gene family encodes a group of proteins which function as transcription factors under physiological conditions and, if aberrantly expressed, can cause cellular transformation. We have recently identified two regulatory elements in the murine immunoglobulin heavy-chain (IgH) enhancer, pi and microB, which exhibit striking similarity to binding sites for ets-related proteins. To identify ets-related transcriptional regulators expressed in pre-B lymphocytes that may interact with either the pi or the microB site, we have used a PCR approach with degenerate oligonucleotides encoding conserved sequences in all members of the ets family. We have cloned the gene for a new ets-related transcription factor, ERP (ets-related protein), from the murine pre-B cell line BASC 6C2 and from mouse lung tissue. The ERP protein contains a region of high homology with the ETS DNA-binding domain common to all members of the ets transcription factor/oncoprotein family. Three additional smaller regions show homology to the ELK-1 and SAP-1 genes, a subgroup of the ets gene family that interacts with the serum response factor. Full-length ERP expresses only negligible DNA-binding activity by itself. Removal of the carboxy terminus enables ERP to interact with a variety of ets-binding sites including the E74 site, the IgH enhancer pi site, and the lck promoter ets site, suggesting a carboxy-terminal negative regulatory domain. At least three ERP-related transcripts are expressed in a variety of tissues. However, within the B-cell lineage, ERP is highly expressed primarily at early stages of B-lymphocyte development, and expression declines drastically upon B-cell maturation, correlating with the enhancer activity of the IgH pi site. These data suggest that ERP might play a role in B-cell development and in IgH gene regulation.

  12. Adaptation of penelope Monte Carlo code system to the absorbed dose metrology: characterization of high energy photon beams and calculations of reference dosimeter correction factors

    International Nuclear Information System (INIS)

    Mazurier, J.

    1999-01-01

    This thesis has been performed in the framework of national reference setting-up for absorbed dose in water and high energy photon beam provided with the SATURNE-43 medical accelerator of the BNM-LPRI (acronym for National Bureau of Metrology and Primary standard laboratory of ionising radiation). The aim of this work has been to develop and validate different user codes, based on PENELOPE Monte Carlo code system, to determine the photon beam characteristics and calculate the correction factors of reference dosimeters such as Fricke dosimeters and graphite calorimeter. In the first step, the developed user codes have permitted the influence study of different components constituting the irradiation head. Variance reduction techniques have been used to reduce the calculation time. The phase space has been calculated for 6, 12 and 25 MV at the output surface level of the accelerator head, then used for calculating energy spectra and dose distributions in the reference water phantom. Results obtained have been compared with experimental measurements. The second step has been devoted to develop an user code allowing calculation correction factors associated with both BNM-LPRI's graphite and Fricke dosimeters thanks to a correlated sampling method starting with energy spectra obtained in the first step. Then the calculated correction factors have been compared with experimental and calculated results obtained with the Monte Carlo EGS4 code system. The good agreement, between experimental and calculated results, leads to validate simulations performed with the PENELOPE code system. (author)

  13. Functional characterization of Bombyx mori nucleopolyhedrovirus late gene transcription and genome replication factors in the non-permissive insect cell line SF-21

    International Nuclear Information System (INIS)

    Berretta, Marcelo F.; Deshpande, Mandar; Crouch, Erin A.; Passarelli, A. Lorena

    2006-01-01

    We compared the abilities of late gene transcription and DNA replication machineries of the baculoviruses Autographa californica nucleopolyhedrovirus (AcMNPV) and Bombyx mori NPV (BmNPV) in SF-21 cells, an insect-derived cell line permissive for AcMNPV infection. It has been well established that 19 AcMNPV late expression factors (lefs) stimulate substantial levels of late gene promoter activity in SF-21 cells. Thus, we constructed a set of clones containing the BmNPV homologs of the AcMNPV lefs under control of the constitutive Drosophila heat shock 70 protein promoter and tested their ability to activate an AcMNPV late promoter-reporter gene cassette in SF-21 cells. We tested the potential of individual or predicted functional groups of BmNPV lefs to successfully replace the corresponding AcMNPV gene(s) in transient late gene expression assays. We found that most, but not all, BmNPV lefs were able to either fully or partially substitute for the corresponding AcMNPV homolog in the context of the remaining AcMNPV lefs with the exception of BmNPV p143, ie-2, and p35. BmNPV p143 was unable to support late gene expression or be imported into the nucleus of cells in the presence of the AcMNPV or the BmNPV LEF-3, a P143 nuclear shuttling factor. Our results suggest that host-specific factors may affect the function of homologous proteins

  14. Biochemical characterization of individual human glycosylated pro-insulin-like growth factor (IGF)-II and big-IGF-II isoforms associated with cancer.

    Science.gov (United States)

    Greenall, Sameer A; Bentley, John D; Pearce, Lesley A; Scoble, Judith A; Sparrow, Lindsay G; Bartone, Nicola A; Xiao, Xiaowen; Baxter, Robert C; Cosgrove, Leah J; Adams, Timothy E

    2013-01-04

    Insulin-like growth factor II (IGF-II) is a major embryonic growth factor belonging to the insulin-like growth factor family, which includes insulin and IGF-I. Its expression in humans is tightly controlled by maternal imprinting, a genetic restraint that is lost in many cancers, resulting in up-regulation of both mature IGF-II mRNA and protein expression. Additionally, increased expression of several longer isoforms of IGF-II, termed "pro" and "big" IGF-II, has been observed. To date, it is ambiguous as to what role these IGF-II isoforms have in initiating and sustaining tumorigenesis and whether they are bioavailable. We have expressed each individual IGF-II isoform in their proper O-glycosylated format and established that all bind to the IGF-I receptor and both insulin receptors A and B, resulting in their activation and subsequent stimulation of fibroblast proliferation. We also confirmed that all isoforms are able to be sequestered into binary complexes with several IGF-binding proteins (IGFBP-2, IGFBP-3, and IGFBP-5). In contrast to this, ternary complex formation with IGFBP-3 or IGFBP-5 and the auxillary protein, acid labile subunit, was severely diminished. Furthermore, big-IGF-II isoforms bound much more weakly to purified ectodomain of the natural IGF-II scavenging receptor, IGF-IIR. IGF-II isoforms thus possess unique biological properties that may enable them to escape normal sequestration avenues and remain bioavailable in vivo to sustain oncogenic signaling.

  15. Isolation and molecular characterization of a novel WIN1/SHN1 ethylene-responsive transcription factor TdSHN1 from durum wheat (Triticum turgidum. L. subsp. durum).

    Science.gov (United States)

    Djemal, Rania; Khoudi, Habib

    2015-11-01

    Over the last decade, APETALA2/Ethylene Responsive Factor (AP2/ERF) proteins have become the subject of intensive research activity due to their involvement in a variety of biological processes. This research led to the identification of AP2/ERF genes in many species; however, little is known about these genes in durum wheat, one of the most important cereal crops in the world. In this study, a new member of the AP2/ERF transcription factor family, designated TdSHN1, was isolated from durum wheat using thermal asymetric interlaced PCR (TAIL-PCR) method. Protein sequence analysis showed that TdSHN1 contained an AP2/ERF domain of 63 amino acids and a putative nuclear localization signal (NLS). Phylogenetic analysis showed that TdSHN1 belongs to a group Va protein in the ERF subfamily which contains the Arabidopsis ERF proteins (SHN1, SHN2, and SHN3). Expression of TdSHN1 was strongly induced by salt, drought, abscisic acid (ABA), and cold. In planta, TdSHN1 protein was able to activate the transcription of GUS reporter gene driven by the GCC box and DRE element sequences. In addition, TdSHN1 was targeted to the nucleus when transiently expressed in tobacco epidermal cells. In transgenic yeast, overexpression of TdSHN1 increased tolerance to multiple abiotic stresses. Taken together, the results showed that TdSHN1 encodes an abiotic stress-inducible, transcription factor which confers abiotic stress tolerance in yeast. TdSHN1 is therefore a promising candidate for improvement of biotic and abiotic stress tolerance in wheat as well as other crops.

  16. Biochemical Characterization of Individual Human Glycosylated pro-Insulin-like Growth Factor (IGF)-II and big-IGF-II Isoforms Associated with Cancer

    Science.gov (United States)

    Greenall, Sameer A.; Bentley, John D.; Pearce, Lesley A.; Scoble, Judith A.; Sparrow, Lindsay G.; Bartone, Nicola A.; Xiao, Xiaowen; Baxter, Robert C.; Cosgrove, Leah J.; Adams, Timothy E.

    2013-01-01

    Insulin-like growth factor II (IGF-II) is a major embryonic growth factor belonging to the insulin-like growth factor family, which includes insulin and IGF-I. Its expression in humans is tightly controlled by maternal imprinting, a genetic restraint that is lost in many cancers, resulting in up-regulation of both mature IGF-II mRNA and protein expression. Additionally, increased expression of several longer isoforms of IGF-II, termed “pro” and “big” IGF-II, has been observed. To date, it is ambiguous as to what role these IGF-II isoforms have in initiating and sustaining tumorigenesis and whether they are bioavailable. We have expressed each individual IGF-II isoform in their proper O-glycosylated format and established that all bind to the IGF-I receptor and both insulin receptors A and B, resulting in their activation and subsequent stimulation of fibroblast proliferation. We also confirmed that all isoforms are able to be sequestered into binary complexes with several IGF-binding proteins (IGFBP-2, IGFBP-3, and IGFBP-5). In contrast to this, ternary complex formation with IGFBP-3 or IGFBP-5 and the auxillary protein, acid labile subunit, was severely diminished. Furthermore, big-IGF-II isoforms bound much more weakly to purified ectodomain of the natural IGF-II scavenging receptor, IGF-IIR. IGF-II isoforms thus possess unique biological properties that may enable them to escape normal sequestration avenues and remain bioavailable in vivo to sustain oncogenic signaling. PMID:23166326

  17. Functional characterization of TRICHOMELESS2, a new single-repeat R3 MYB transcription factor in the regulation of trichome patterning in Arabidopsis

    Directory of Open Access Journals (Sweden)

    Gan Lijun

    2011-12-01

    Full Text Available Abstract Background Single-repeat R3 MYB transcription factors (single-repeat MYBs play important roles in controlling trichome patterning in Arabidopsis. It was proposed that single-repeat MYBs negatively regulate trichome formation by competing with GLABRA1 (GL1 for binding GLABRA3/ENHANCER OF GLABRA3 (GL3/EGL3, thus inhibiting the formation of activator complex TTG1(TRANSPARENT TESTA GLABRA1-GL3/EGL3-GL1 that is required for the activation of GLABRA2 (GL2, whose product is a positive regulator of trichome formation. Previously we identified a novel single-repeat MYB transcription factor, TRICHOMELESS1 (TCL1, which negatively regulates trichome formation on the inflorescence stems and pedicels by directly suppressing the expression of GL1. Results We analyzed here the role of TRICHOMELESS2 (TCL2, a previously-uncharacterized single-repeat MYB transcription factor in trichome patterning in Arabidopsis. We showed that TCL2 is closely related to TCL1, and like TCL1 and other single-repeat MYBs, TCL2 interacts with GL3. Overexpression of TCL2 conferred glabrous phenotype while knockdown of TCL2 via RNAi induced ectopic trichome formation on the inflorescence stems and pedicels, a phenotype that was previously observed in tcl1 mutants. These results suggested that TCL2 may have overlapping function with TCL1 in controlling trichome formation on inflorescences. On the other hand, although the transcription of TCL2, like TCL1, is not controlled by the activator complex formed by GL1 and GL3, and TCL2 and TCL1 proteins are more than 80% identical at the amino acid level, the expression of TCL2 under the control of TCL1 promoter only partially recovered the mutant phenotype of tcl1, implying that TCL2 and TCL1 are not fully functional equivalent. Conclusions TCL2 function redundantly with TCL1 in controlling trichome formation on inflorescences, but they are not fully functional equivalent. Transcription of TCL2 is not controlled by activator complex

  18. Characterization of the growth of murine fibroblasts that express human insulin receptors. I. The effect of insulin in the absence of other growth factors

    International Nuclear Information System (INIS)

    Randazzo, P.A.; Morey, V.A.; Polishook, A.K.; Jarett, L.

    1990-01-01

    The effect of insulin on the growth of murine fibroblasts transfected with an expression vector containing human insulin receptor cDNA (NIH 3T3/HIR) and the parental cells (NIH/3T3) was characterized. Insulin in the absence of other mitogens increased the rate of incorporation of thymidine into NIH 3T3/HIR cells with a half-maximal response occurring at an insulin concentration of 35 ng/ml and a maximal response that was equivalent to that elicited by 10% fetal calf serum. The thymidine incorporation rate was increased by 12 h, was maximal at approximately 16 h, and returned to basal rates at 24 h after the addition of insulin. Insulin induced a maximum of 65% of cells to incorporate thymidine. The increased DNA synthesis was accompanied by net growth. Addition of insulin to the NIH 3T3/HIR cells resulted in increased DNA content with a half-maximal response occurring at approximately 30 ng/ml insulin and a maximal response equivalent to that elicited by serum. An increase in cell number detected after the addition of insulin to the NIH 3T3/HIR suggests that the cells had progressed through mitosis. Insulin did not increase the rate of thymidine incorporation, DNA content, or number of the parental NIH 3T3 cells. These data show that insulin, in the absence of a second mitogen, is able to induce NIH 3T3/HIR fibroblasts to traverse the cell cycle

  19. SAPO-34/AlMCM-41, as a novel hierarchical nanocomposite: preparation, characterization and investigation of synthesis factors using response surface methodology

    Science.gov (United States)

    Roohollahi, Hossein; Halladj, Rouein; Askari, Sima; Yaripour, Fereydoon

    2018-06-01

    SAPO-34/AlMCM-41, as a new hierarchical nanocomposite was successfully synthesized via hydrothermal and dry-gel conversion. In an experimental and statistical study, effect of five input parameters including synthesis period, drying temperature, NaOH/Si, water/dried-gel and SAPO% were investigated on range-order degree of mesochannels and the relative crystallinity. X-ray diffraction (XRD) patterns were recorded to characterize the ordered AlMCM-41 and crystalline SAPO-34 structures. Nitrogen adsorption-desorption technique, scanning electron microscopy (SEM), field-emission SEM (FESEM) equipped with an energy-dispersive X-ray spectroscopy (EDS-Map) and transmission electron microscopy (TEM) were used to study the textural properties, morphology and surface elemental composition. Two reduced polynomials were fitted to the responses with good precision. Further, based on analysis of variances, SAPO% and time duration of dry-gel conversion were observed as the most effective parameters on the composite structure. The hierarchical porosity, narrow pore size distribution, high external surface area and large specific pore volume were of interesting characteristics for this novel nanocomposite.

  20. Molecular characterization of the Jatropha curcas JcR1MYB1 gene encoding a putative R1-MYB transcription factor

    Directory of Open Access Journals (Sweden)

    Hui-Liang Li

    2014-09-01

    Full Text Available The cDNA encoding the R1-MYB transcription factor, designated as JcR1MYB1, was isolated from Jatropha curcas using rapid amplification of cDNA ends. JcR1MYB1 contains a 951 bp open reading frame that encodes 316 amino acids. The deduced JcR1MYB1 protein was predicted to possess the conserved, 56-amino acid-long DNA-binding domain, which consists of a single helix-turn-helix module and usually occurs in R1-MYBs. JcR1MYB1 is a member of the R1-MYB transcription factor subfamily. A subcellular localization study confirmed the nuclear localization of JcR1MYB1. Expression analysis showed that JcR1MYB1 transcripts accumulated in various examined tissues, with high expression levels in the root and low levels in the stem. JcR1MYB1 transcription was up-regulated by polyethylene glycol, NaCl, and cold treatments, as well as by abscisic acid, jasmonic acid, and ethylene treatment. Analysis of transgenic tobacco plants over-expressing JcR1MYB1 indicates an inportant function for this gene in salt stress.

  1. Successive ion layer adsorption and reaction (SILAR) technique synthesis of Al(III)-8-hydroxy-5-nitrosoquinolate nano-sized thin films: characterization and factors optimization.

    Science.gov (United States)

    Haggag, Sawsan M S; Farag, A A M; Abdel Refea, M

    2013-02-01

    Nano Al(III)-8-hydroxy-5-nitrosoquinolate [Al(III)-(HNOQ)(3)] thin films were synthesized by the rapid, direct, simple and efficient successive ion layer adsorption and reaction (SILAR) technique. Thin film formation optimized factors were evaluated. Stoichiometry and structure were confirmed by elemental analysis and FT-IR. The particle size (27-71 nm) was determined using scanning electron microscope (SEM). Thermal stability and thermal parameters were determined by thermal gravimetric analysis (TGA). Optical properties were investigated using spectrophotometric measurements of transmittance and reflectance at normal incidence. Refractive index, n, and absorption index, k, were determined. Spectral behavior of the absorption coefficient in the intrinsic absorption region revealed a direct allowed transition with 2.45 eV band gap. The current-voltage (I-V) characteristics of [Al(III)-(HNOQ)(3)]/p-Si heterojunction was measured at room temperature. The forward and reverse I-V characteristics were analyzed. The calculated zero-bias barrier height (Φ(b)) and ideality factor (n) showed strong bias dependence. Energy distribution of interface states (N(ss)) was obtained. Copyright © 2012 Elsevier B.V. All rights reserved.

  2. Molecular cloning and characterization of a novel freezing-inducible DREB1/CBF transcription factor gene in boreal plant Iceland poppy (Papaver nudicaule

    Directory of Open Access Journals (Sweden)

    Zhuo Huang

    Full Text Available Abstract DREB1 of the AP2/ERF superfamily plays a key role in the regulation of plant response to low temperatures. In this study, a novel DREB1/CBF transcription factor, PnDREB1, was isolated from Iceland poppy (Papaver nudicaule, a plant adaptive to low temperature environments. It is homologous to the known DREB1s of Arabidopsis and other plant species. It also shares similar 3D structure, and conserved and functionally important motifs with DREB1s of Arabidopsis. The phylogenetic analysis indicated that the AP2 domain of PnDREB1 is similar to those of Glycine max, Medicago truncatula, and M. sativa. PnDREB1 is constitutively expressed in diverse tissues and is increased in roots. qPCR analyses indicated that PnDREB1 is significantly induced by freezing treatment as well as by abscissic acid. The expression levels induced by freezing treatment were higher in the variety with higher degree of freezing tolerance. These results suggested that PnDREB1 is a novel and functional DREB1 transcription factor involved in freezing response and possibly in other abiotic stresses. Furthermore, the freezing-induction could be suppressed by exogenous gibberellins acid, indicating that PnDREB1 might play some role in the GA signaling transduction pathway. This study provides a basis for better understanding the roles of DREB1 in adaption of Iceland poppy to low temperatures.

  3. Tentative characterization of precursor compounds and co-factors of pigment formation in production of 'wu mi' from Vaccinium bracteatum Thunb. Leaves.

    Science.gov (United States)

    Fan, Mingcong; Fan, Yihui; Huang, Weiping; Wang, Li; Li, Yan; Qian, Haifeng; Zhang, Hui; Qi, Xiguang

    2018-10-01

    Vaccinium bracteatum leaves (VBTL) are traditionally used in China to dye rice grains, which assume a deep blue color, named 'Wu mi'. Information on the mechanism of pigment formation is limited. In this study, CIELAB color space parameters were used to represent the color of 'Wu mi'. Precursor compounds of pigments formed during the dyeing process were identified by UPLC Q-TOF MS analysis. The changes in co-factors for pigment formation in VBTL were measured at different growth stages. The L ∗ and b ∗ values of dyed rice increased as the leaves aged, whereas a ∗ values showed irregular changes. Six compounds were tentatively identified as pigment precursors by UPLC Q-TOF MS analysis. The pH and β-glucosidase activity at different growth stages of VBTL were indicated to be crucial co-factors for pigment formation. A tentative hypothesis is presented that iridoid glycosides are hydrolyzed by acids and β-glucosidases to form a dialdehyde structure that binds covalently with amino residues of lysine side chains in rice protein molecules. Copyright © 2018 Elsevier Ltd. All rights reserved.

  4. Caracterización de factores clínico-epidemiológicos en la retinopatía del prematuro Characterization of clinical-epidemiological factors present in premature retinopathy

    Directory of Open Access Journals (Sweden)

    Angelicia Crespo Campos

    2009-09-01

    Full Text Available INTRODUCCIÓN. La retina en desarrollo es muy sensible a toda clase de alteraciones en el aporte de oxígeno. La presente investigación buscó evaluar los factores clínico-epidemiológicos relacionados con dicha entidad en nuestro medio. MÉTODOS. Se realizó una investigación observacional, longitudinal, ambispectiva y descriptiva, en la Unidad de Cuidados Intensivos del Servicio de Neonatología del Hospital «Abel Santamaría», en el período comprendido entre los años 2001 y 2007. Se estudiaron los 48 recién nacidos con diagnóstico de retinopatía. Se analizaron las variables: sexo, edad gestacional, peso, crecimiento intrauterino, vía de nacimiento, antecedentes maternos, necesidad de ventilación u oxigenación y morbilidad asociada. Las variables se analizaron mediante pruebas de comparación de dos y k proporciones de grupos independientes. RESULTADOS. Se identificaron como principales factores de riesgo para el padecimiento de la retinopatía de la prematuridad: el sexo masculino, la edad gestacional por debajo de 31,6 semanas, el peso menor de 1499 g, la necesidad de ventilación mecánica u oxigenoterapia por más de 96 h, así como las patologías respiratorias y la sepsis. CONCLUSIONES. La enfermedad es compleja y su causa, probablemente multifactorial.INTRODUCTION: Developing retina is very sensitive to all types de alterations in oxygen support. Aim of present research was to assess the clinical-pathological factors related to such entity in our practice. METHODS: An observational, longitudinal, ambispective and descriptive research was conducted in Intensive Care Unit from Neonatology Service of "Abel Santamaría Cuadrado" Hospital during 2001 and 2007. A total of 48 newborn diagnosed with retinopathy was studied. Variables analyzed included sex, age, gestational age, weight, intrauterine development, birth route, maternal backgrounds, ventilation or oxygenation dependence, and associated morbidity using two

  5. Characterization of the effect of serum and chelating agents on Staphylococcus aureus biofilm formation; chelating agents augment biofilm formation through clumping factor B

    Science.gov (United States)

    Abraham, Nabil Mathew

    Staphylococcus aureus is the causative agent of a diverse array of acute and chronic infections, and some these infections, including infective endocarditis, joint infections, and medical device-associated bloodstream infections, depend upon its capacity to form tenacious biofilms on surfaces. Inserted medical devices such as intravenous catheters, pacemakers, and artificial heart valves save lives, but unfortunately, they can also serve as a substrate on which S. aureus can form a biofilm, attributing S. aureus as a leading cause of medical device-related infections. The major aim of this work was take compounds to which S. aureus would be exposed during infection and to investigate their effects on its capacity to form a biofilm. More specifically, the project investigated the effects of serum, and thereafter of catheter lock solutions on biofilm formation by S. aureus. Pre-coating polystyrene with serum is frequently used as a method to augment biofilm formation. The effect of pre-coating with serum is due to the deposition of extracellular matrix components onto the polystyrene, which are then recognized by MSCRAMMs. We therefore hypothesized that the major component of blood, serum, would induce biofilm formation. Surprisingly, serum actually inhibited biofilm formation. The inhibitory activity was due to a small molecular weight, heat-stable, non-proteinaceous component/s of serum. Serum-mediated inhibition of biofilm formation may represent a previously uncharacterized aspect of host innate immunity that targets the expression of a key bacterial virulence factor: the ability to establish a resistant biofilm. Metal ion chelators like sodium citrate are frequently chosen to lock intravenous catheters because they are regarded as potent inhibitors of bacterial biofilm formation and viability. We found that, while chelating compounds abolished biofilm formation in most strains of S. aureus, they actually augmented the phenotype in a subset of strains. We

  6. Genome Wide Identification and Characterization of Apple bHLH Transcription Factors and Expression Analysis in Response to Drought and Salt Stress.

    Science.gov (United States)

    Mao, Ke; Dong, Qinglong; Li, Chao; Liu, Changhai; Ma, Fengwang

    2017-01-01

    The bHLH (basic helix-loop-helix) transcription factor family is the second largest in plants. It occurs in all three eukaryotic kingdoms, and plays important roles in regulating growth and development. However, family members have not previously been studied in apple. Here, we identified 188 MdbHLH proteins in apple "Golden Delicious" ( Malus × domestica Borkh.), which could be classified into 18 groups. We also investigated the gene structures and 12 conserved motifs in these MdbHLH s. Coupled with expression analysis and protein interaction network prediction, we identified several genes that might be responsible for abiotic stress responses. This study provides insight and rich resources for subsequent investigations of such proteins in apple.

  7. Identification and characterization of finger millet OPAQUE2 transcription factor gene under different nitrogen inputs for understanding their role during accumulation of prolamin seed storage protein.

    Science.gov (United States)

    Gaur, Vikram Singh; Kumar, Lallan; Gupta, Supriya; Jaiswal, J P; Pandey, Dinesh; Kumar, Anil

    2018-03-01

    In this study, we report the isolation and characterization of the mRNA encoding OPAQUE2 (O2) like TF of finger millet (FM) ( Eleusine coracana) ( EcO2 ). Full-length EcO2 mRNA was isolated using conserved primers designed by aligning O2 mRNAs of different cereals followed by 3' and 5' RACE (Rapid Amplification of cDNA Ends). The assembled full-length EcO2 mRNA was found to contain an ORF of 1248-nt coding the 416 amino acids O2 protein. Domain analysis revealed the presence of the BLZ and bZIP-C domains which is a characteristic feature of O2 proteins. Phylogenetic analysis of EcO2 protein with other bZIP proteins identified using finger millet transcriptome data and O2 proteins of other cereals showed that EcO2 shared high sequence similarity with barley BLZ1 protein. Transcripts of EcO2 were detected in root, stem, leaves, and seed development stages. Furthermore, to investigate nitrogen responsiveness and the role of EcO2 in regulating seed storage protein gene expression, the expression profiles of EcO2 along with an α-prolamin gene were studied during the seed development stages of two FM genotypes (GE-3885 and GE-1437) differing in grain protein content (13.8 and 6.2%, respectively) grown under increasing nitrogen inputs. Compared to GE-1437, the EcO2 was relatively highly expressed during the S2 stage of seed development which further increased as nitrogen input was increased. The Ecα - prolamin gene was strongly induced in the high protein genotype (GE-3885) at all nitrogen inputs. These results indicate the presence of nitrogen responsiveness regulatory elements which might play an important role in accumulating protein in FM genotypes through modulating EcO2 expression by sensing plant nitrogen status.

  8. Molecular characterization of BrMYB28 and BrMYB29 paralogous transcription factors involved in the regulation of aliphatic glucosinolate profiles in Brassica rapa ssp. pekinensis.

    Science.gov (United States)

    Baskar, Venkidasamy; Park, Se Won

    2015-07-01

    Glucosinolates (GSL) are one of the major secondary metabolites of the Brassicaceae family. In the present study, we aim at characterizing the multiple paralogs of aliphatic GSL regulators, such as BrMYB28 and BrMYB29 genes in Brassica rapa ssp. pekinensis, by quantitative real-time PCR (qRT-PCR) analysis in different tissues and at various developmental stages. An overlapping gene expression pattern between the BrMYBs as well as their downstream genes (DSGs) was found at different developmental stages. Among the BrMYB28 and BrMYB29 paralogous genes, the BrMYB28.3 and BrMYB29.1 genes were dominantly expressed in most of the developmental stages, compared to the other paralogs of the BrMYB genes. Furthermore, the differential expression pattern of the BrMYBs was observed under various stress treatments. Interestingly, BrMYB28.2 showed the least expression in most developmental stages, while its expression was remarkably high in different stress conditions. More specifically, the BrMYB28.2, BrMYB28.3, and BrMYB29.1 genes were highly responsive to various abiotic and biotic stresses, further indicating their possible role in stress tolerance. Moreover, the in silico cis motif analysis in the upstream regulatory regions of BrMYBs showed the presence of various putative stress-specific motifs, which further indicated their responsiveness to biotic and abiotic stresses. These observations suggest that the dominantly expressed BrMYBs, both in different developmental stages and under various stress treatments (BrMYB28.3 and BrMYB29.1), may be potential candidate genes for altering the GSL level through genetic modification studies in B. rapa ssp. pekinensis. Copyright © 2015. Published by Elsevier SAS.

  9. Using a minigene approach to characterize a novel splice site mutation in human F7 gene causing inherited factor VII deficiency in a Chinese pedigree.

    Science.gov (United States)

    Yu, T; Wang, X; Ding, Q; Fu, Q; Dai, J; Lu, Y; Xi, X; Wang, H

    2009-11-01

    Factor VII deficiency which transmitted as an autosomal recessive disorder is a rare haemorrhagic condition. The aim of this study was to identify the molecular genetic defect and determine its functional consequences in a Chinese pedigree with FVII deficiency. The proband was diagnosed as inherited coagulation FVII deficiency by reduced plasma levels of FVII activity (4.4%) and antigen (38.5%). All nine exons and their flanking sequence of F7 gene were amplified by polymerase chain reaction (PCR) for the proband and the PCR products were directly sequenced. The compound heterozygous mutations of F7 (NM_000131.3) c.572-1G>A and F7 (NM_000131.3) c.1165T>G; p.Cys389Gly were identified in the proband's F7 gene. To investigate the splicing patterns associated with F7 c.572-1G>A, ectopic transcripts in leucocytes of the proband were analyzed. F7 minigenes, spanning from intron 4 to intron 7 and carrying either an A or a G at position -1 of intron 5, were constructed and transiently transfected into human embryonic kidney (HEK) 293T cells, followed by RT-PCR analysis. The aberrant transcripts from the F7 c.572-1G>A mutant allele were not detected by ectopic transcription study. Sequencing of the RT-PCR products from the mutant transfectant demonstrated the production of an erroneously spliced mRNA with exon 6 skipping, whereas a normal splicing occurred in the wide type transfectant. The aberrant mRNA produced from the F7 c.572-1G>A mutant allele is responsible for the factor VII deficiency in this pedigree.

  10. Phenotypic and molecular characterization of antimicrobial resistance and virulence factors in Pseudomonas aeruginosa clinical isolates from Recife, State of Pernambuco, Brazil

    Directory of Open Access Journals (Sweden)

    Paula Regina Luna de Araújo Jácome

    2012-12-01

    Full Text Available INTRODUCTION: The emergence of carbapenem resistance mechanisms in Pseudomonas aeruginosa has been outstanding due to the wide spectrum of antimicrobial degradation of these bacteria, reducing of therapeutic options. METHODS: Sixty-one clinical strains of P. aeruginosa isolated from five public hospitals in Recife, Pernambuco, Brazil, were examined between 2006 and 2010, aiming of evaluating the profiles of virulence, resistance to antimicrobials, presence of metallo-β-lactamase (MBL genes, and clonal relationship among isolates. RESULTS: A high percentage of virulence factors (34.4% mucoid colonies; 70.5% pyocyanin; 93.4% gelatinase positives; and 72.1% hemolysin positive and a high percentage of antimicrobial resistance rates (4.9% pan-resistant and 54.1% multi-drug resistant isolates were observed. Among the 29 isolates resistant to imipenem and/or ceftazidime, 44.8% (13/29 were MBL producers by phenotypic evaluation, and of these, 46.2% (6/13 were positive for the blaSPM-1 gene. The blaIMP and blaVIM genes were not detected. The molecular typing revealed 21 molecular profiles of which seven were detected in distinct hospitals and periods. Among the six positive blaSPM-1 isolates, three presented the same clonal profile and were from the same hospital, whereas the other three presented different clonal profiles. CONCLUSIONS: These results revealed that P. aeruginosa is able to accumulate different resistance and virulence factors, making the treatment of infections difficult. The identification of blaSPM-1 genes and the dissemination of clones in different hospitals, indicate the need for stricter application of infection control measures in hospitals in Recife, Brazil, aiming at reducing costs and damages caused by P. aeruginosa infections.

  11. Characterization of a human coagulation factor Xa-binding site on Viperidae snake venom phospholipases A2 by affinity binding studies and molecular bioinformatics

    Directory of Open Access Journals (Sweden)

    Gowda Veerabasappa T

    2007-12-01

    Full Text Available Abstract Background The snake venom group IIA secreted phospholipases A2 (SVPLA2, present in the Viperidae snake family exhibit a wide range of toxic and pharmacological effects. They exert their different functions by catalyzing the hydrolysis of phospholipids (PL at the membrane/water interface and by highly specific direct binding to: (i presynaptic membrane-bound or intracellular receptors; (ii natural PLA2-inhibitors from snake serum; and (iii coagulation factors present in human blood. Results Using surface plasmon resonance (SPR protein-protein interaction measurements and an in vitro biological test of inhibition of prothrombinase activity, we identify a number of Viperidae venom SVPLA2s that inhibit blood coagulation through direct binding to human blood coagulation factor Xa (FXa via a non-catalytic, PL-independent mechanism. We classify the SVPLA2s in four groups, depending on the strength of their binding. Molecular electrostatic potentials calculated at the surface of 3D homology-modeling models show a correlation with inhibition of prothrombinase activity. In addition, molecular docking simulations between SVPLA2 and FXa guided by the experimental data identify the potential FXa binding site on the SVPLA2s. This site is composed of the following regions: helices A and B, the Ca2+ loop, the helix C-β-wing loop, and the C-terminal fragment. Some of the SVPLA2 binding site residues belong also to the interfacial binding site (IBS. The interface in FXa involves both, the light and heavy chains. Conclusion We have experimentally identified several strong FXa-binding SVPLA2s that disrupt the function of the coagulation cascade by interacting with FXa by the non-catalytic PL-independent mechanism. By theoretical methods we mapped the interaction sites on both, the SVPLA2s and FXa. Our findings may lead to the design of novel, non-competitive FXa inhibitors.

  12. Metallogenic epoch of the Jiapigou gold belt, Jilin Province, China ...

    Indian Academy of Sciences (India)

    29, Xueyuan Road, Beijing 100083, People's Republic of China. 2Beijing Research .... terrane, which is separated from the central Asian orogenic belt .... Stage I formed the main body of quartz veins ...... size (20–25 μm) fluid inclusions can be observed, although most ..... Canada and Western Australia (Goldfarb et al. 2001 ...

  13. Cloning and characterization of a novel stress-responsive WRKY transcription factor gene (MusaWRKY71) from Musa spp. cv. Karibale Monthan (ABB group) using transformed banana cells.

    Science.gov (United States)

    Shekhawat, Upendra K Singh; Ganapathi, Thumballi R; Srinivas, Lingam

    2011-08-01

    WRKY transcription factor proteins play significant roles in plant stress responses. Here, we report the cloning and characterization of a novel WRKY gene, MusaWRKY71 isolated from an edible banana cultivar Musa spp. Karibale Monthan (ABB group). MusaWRKY71, initially identified using in silico approaches from an abiotic stress-related EST library, was later extended towards the 3' end using rapid amplification of cDNA ends technique. The 1299-bp long cDNA of MusaWRKY71 encodes a protein with 280 amino acids and contains a characteristic WRKY domain in the C-terminal half. Although MusaWRKY71 shares good similarity with other monocot WRKY proteins the substantial size difference makes it a unique member of the WRKY family in higher plants. The 918-bp long 5' proximal region determined using thermal asymmetric interlaced-polymerase chain reaction has many putative cis-acting elements and transcription factor binding motifs. Subcellular localization assay of MusaWRKY71 performed using a GFP-fusion platform confirmed its nuclear targeting in transformed banana suspension cells. Importantly, MusaWRKY71 expression in banana plantlets was up-regulated manifold by cold, dehydration, salt, ABA, H2O2, ethylene, salicylic acid and methyl jasmonate treatment indicating its involvement in response to a variety of stress conditions in banana. Further, transient overexpression of MusaWRKY71 in transformed banana cells led to the induction of several genes, homologues of which have been proven to be involved in diverse stress responses in other important plants. The present study is the first report on characterization of a banana stress-related transcription factor using transformed banana cells.

  14. Molecular characterization of organic matter in converted forests in Western Europe; disentangling the effects of edaphic factors and input differences on SOM composition

    Science.gov (United States)

    Brock, Olaf; Kooijman, Annemieke; Vancampenhout, Karen; Muys, Bart; Jansen, Boris

    2017-04-01

    By storing carbon in the soil, forests can play an important role in climate mitigation. We studied how the SOM composition was affected by conversion of deciduous stands to mono-culture spruce plantations in the Mullerthal in Luxembourg and the Gaume in south-east Belgium. Both regions have a known and similar vegetation history on different lithologies, ranging from carcareous marls to decalcified sands. Lignin and cutin/suberin biomarkers were identified by using thermally assisted hydrolysis and methylation (THM) with unlabelled tetra methyl ammonium hydroxide (TMAH). Lignin was used to distinguish deciduous and coniferous litter sources, whereas cutin and suberin indicated the respective input of above- and belowground litter input. A twinplot setup was used to be able to independently evaluate the effect of edaphic factors versus input differences on SOM composition. pH values and SOC stocks reflected the lithological gradients in both study areas. The difference was more subtle in the Gaume where the gradient is much narrower. The existence of pedogenic thresholds explains why significant differences in lignin yield and SOC stocks between plots with different lithology were also found along the subtle gradient in the Gaume. Secondly, we observed differences in molecular composition and also in decomposition state of lignin that were caused solely by input differences between adjacent deciduous and coniferous forest plots. Furthermore, we found a legacy effect, a signal of former deciduous forest in the deeper soil layers (15-20 cm) under the current spruce plantations, in the loamy substrate plots of the Gaume, which was not observed in the Mullerthal, despite the similar vegetation history of both regions. This can be explained by differences in environmental conditions between both areas. Higher pH values resulting in a higher biological activity could explain the absence of a legacy effect in the Mullerthal plots. Therefore, an important conclusion of

  15. Establishment and characterization of cetuximab resistant head and neck squamous cell carcinoma cell lines: focus on the contribution of the AP-1 transcription factor

    Science.gov (United States)

    Boeckx, Carolien; Blockx, Lina; de Beeck, Ken Op; Limame, Ridha; Camp, Guy Van; Peeters, Marc; Vermorken, Jan B; Specenier, Pol; Wouters, An; Baay, Marc; Lardon, Filip

    2015-01-01

    Background: After an initial response to EGFR targeted therapy, secondary resistance almost invariably ensues, thereby limiting the clinical benefit of the drug. Hence, it has been recognized that the successful implementation of targeted therapy in the treatment of HNSCC cancer is very much dependent on predictive biomarkers for patient selection. Methods: We generated an in vitro model of acquired cetuximab resistance by chronically exposing three HNSCC cell lines to increasing cetuximab doses. Gene expression profiles of sensitive parental cells and resistant daughter cells were compared using microarray analysis. Growth inhibitory experiments were performed with an HB-EGF antibody and the MMP inhibitor, both in combination with cetuximab. Characteristics of EMT were analyzed using migration and invasion assays, immunofluorescent vimentin staining and qRT-PCR for several genes involved in this process. The function of the transcription factor AP-1 was investigated using qRT-PCR for several genes upregulated or downregulated in cetuximab resistant cells. Furthermore, anchorage-independent growth was investigated using the soft agar assay. Results: Gene expression profiling shows that cetuximab resistant cells upregulate several genes, including interleukin 8, the EGFR ligand HB-EGF and the metalloproteinase ADAM19. Cytotoxicity experiments with neutralizing HB-EGF antibody could not induce any growth inhibition, whereas an MMP inhibitor inhibited cell growth in cetuximab resistant cells. However, no synergetic effects combined with cetuximab could be observed. Cetuximab resistant cells showed traits of EMT, as witnessed by increased migratory potential, increased invasive potential, increased vimentine expression and increased expression of several genes involved in EMT. Furthermore, expression of upregulated genes could be repressed by the treatment with apigenin. The cetuximab resistant LICR-HN2 R10.3 cells tend to behave differently in cell culture, forming

  16. Characterization of brown adipose tissue 18F-FDG uptake in PET/CT imaging and its influencing factors in the Chinese population

    International Nuclear Information System (INIS)

    Shao, Xiaonan; Shao, Xiaoliang; Wang, Xiaosong; Wang, Yuetao

    2016-01-01

    18 F-FDG PET/CT has been widely applied for tumor imaging. However, it is reported that many normal tissues, e.g., brown adipose tissue, can also uptake 18 F-FDG. The purpose of this study was to determine the imaging characteristics of 18 F-FDG uptake in brown adipose tissue (BAT) in PET/CT. A total of 2,944 patients who underwent PET/CT from September 2011 to March 2013 were analyzed retrospectively. Imaging features of 18 F-FDG uptake in BAT were analyzed. Univariate analysis and logistic regression analysis were performed to evaluate the effect of age, gender, cancer status, body mass index (BMI), average daily maximum temperature of imaging month and fasting plasma glucose (Glu) on the positive rate of 18 F-FDG uptake in BAT. The results showed that 1.9% (57/2944) patients had 18 F-FDG uptake in BAT. 18 F-FDG, manifested as flaky, nodular and beaded shape, was symmetrically distributed in the adipose tissues of cervical and supraclavicular, mediastinal, paravertebral, and perirenal areas. Uptake of 18 F-FDG within cervical/supraclavicular area was most common (89.5%, 51/57) with an SUV max ranging from 2.8 to 31.4. Univariate analysis showed that gender and cancer status were not significantly correlated with the BAT 18 F-FDG uptake rate. In contrast, age, BMI, Glu and average daily maximum temperature in the imaging month were significantly correlated with the BAT 18 F-FDG uptake rate (P < 0.05). Further logistic regression analysis showed that only age, BMI and average daily maximum temperature were significant (OR < 1, P < 0.05). Based on the value of OR, the most significant factor that affects BAT 18 F-FDG uptake rate was age, followed by the average daily maximum temperature and BMI. We concluded that Chinese adult has low positive rate of 18 F-FDG uptake in BAT. Cervical/Supraclavicular is the most common area with BAT 18 F-FDG uptake. Age, average daily maximum temperature and BMI are independent factors affecting 18 F-FDG uptake.

  17. Functional characterization of rs2229094 (T>C polymorphism in the tumor necrosis factor locus and lymphotoxin alpha expression in human retina: the Retina 4 project

    Directory of Open Access Journals (Sweden)

    Pastor-Idoate S

    2017-05-01

    with PVR and its relationship with tumor necrosis factor locus, warrant further studies and could justify the use of this polymorphism as a valid biomarker to identify high-risk patients to develop PVR after RD. Keywords: proliferative vitreoretinopathy, lymphotoxin alpha, tumor necrosis factor alpha, inflammation, cytokines, polymorphism

  18. Seasonal characterization of CDOM for lakes in semiarid regions of Northeast China using excitation-emission matrix fluorescence and parallel factor analysis (EEM-PARAFAC)

    Science.gov (United States)

    Zhao, Ying; Song, Kaishan; Wen, Zhidan; Li, Lin; Zang, Shuying; Shao, Tiantian; Li, Sijia; Du, Jia

    2016-03-01

    The seasonal characteristics of fluorescent components in chromophoric dissolved organic matter (CDOM) for lakes in the semiarid region of Northeast China were examined by excitation-emission matrix (EEM) spectra and parallel factor analysis (PARAFAC). Two humic-like (C1 and C2) and protein-like (C3 and C4) components were identified using PARAFAC. The average fluorescence intensity of the four components differed under seasonal variation from June and August 2013 to February and April 2014. Components 1 and 2 exhibited a strong linear correlation (R2 = 0.628). Significantly positive linear relationships between CDOM absorption coefficients a(254) (R2 = 0.72, 0.46, p DOC). However, almost no obvious correlation was found between salinity and EEM-PARAFAC-extracted components except for C3 (R2 = 0.469). Results from this investigation demonstrate that the EEM-PARAFAC technique can be used to evaluate the seasonal dynamics of CDOM fluorescent components for inland waters in the semiarid regions of Northeast China, and to quantify CDOM components for other waters with similar environmental conditions.

  19. Nicotine can skew the characterization of the macrophage type-1 (MΦ1) phenotype differentiated with granulocyte-macrophage colony-stimulating factor to the MΦ2 phenotype

    International Nuclear Information System (INIS)

    Yanagita, Manabu; Kobayashi, Ryohei; Murakami, Shinya

    2009-01-01

    Macrophages (MΦs) exhibit functional heterogeneity and plasticity in the local microenvironment. Recently, it was reported that MΦs can be divided into proinflammatory MΦs (MΦ1) and anti-inflammatory MΦs (MΦ2) based on their polarized functional properties. Here, we report that nicotine, the major ingredient of cigarette smoke, can modulate the characteristics of MΦ1. Granulocyte-macrophage colony-stimulating factor-driven MΦ1 with nicotine (Ni-MΦ1) showed the phenotypic characteristics of MΦ2. Like MΦ2, Ni-MΦ1 exhibited antigen-uptake activities. Ni-MΦ1 suppressed IL-12, but maintained IL-10 and produced high amounts of MCP-1 upon lipopolysaccharide stimulation compared with MΦ1. Moreover, we observed strong proliferative responses of T cells to lipopolysaccharide-stimulated MΦ1, whereas Ni-MΦ1 reduced T cell proliferation and inhibited IFN-γ production by T cells. These results suggest that nicotine can change the functional characteristics of MΦ and skew the MΦ1 phenotype to MΦ2. We propose that nicotine is a potent regulator that modulates immune responses in microenvironments.

  20. High Surface-Enhanced Raman Scattering (SERS) Amplification Factor Obtained with Silver Printed Circuit Boards and the Influence of Phenolic Resins for the Characterization of the Pesticide Thiram.

    Science.gov (United States)

    Silva de Almeida, Francylaine; Bussler, Larissa; Marcio Lima, Sandro; Fiorucci, Antonio Rogério; da Cunha Andrade, Luis Humberto

    2016-07-01

    In this work, low-cost substrates with rough silver surfaces were prepared from commercial copper foil-covered phenolic board (CPB) and an aqueous solution of AgNO3, and were used for surface-enhanced Raman scattering (SERS) and surface-enhanced resonance Raman scattering (SERRS) measurements. A maximum SERS amplification factor of 1.2 × 10(7) was obtained for Rhodamine 6G (R6G), and use of the CPB resulted in a detection limit for Thiram pesticide of 0.5 µmol L(-1) The minimum detection level was limited by residual traces of phenolic groups that originated from the substrate resin, which became solubilized in the aqueous Ag(+) solution. It was found that the bands corresponding to the impurities had less influence in the Thiram analysis, which could be explained by the high affinity of sulfur for Ag surfaces. The influence of impurities in the SERS analyses therefore depended on the linkage between the rough silver surface and the analyte. The findings demonstrated the ease and effectiveness of using CPB to prepare a nanostructured surface for SERS. © The Author(s) 2016.

  1. Biochemical and redox characterization of the mediator complex and its associated transcription factor GeBPL, a GLABROUS1 enhancer binding protein.

    Science.gov (United States)

    Shaikhali, Jehad; Davoine, Céline; Brännström, Kristoffer; Rouhier, Nicolas; Bygdell, Joakim; Björklund, Stefan; Wingsle, Gunnar

    2015-06-15

    The eukaryotic mediator integrates regulatory signals from promoter-bound transcription factors (TFs) and transmits them to RNA polymerase II (Pol II) machinery. Although redox signalling is important in adjusting plant metabolism and development, nothing is known about a possible redox regulation of mediator. In the present study, using pull-down and yeast two-hybrid assays, we demonstrate the association of mediator (MED) subunits MED10a, MED28 and MED32 with the GLABROUS1 (GL1) enhancer-binding protein-like (GeBPL), a plant-specific TF that binds a promoter containing cryptochrome 1 response element 2 (CryR2) element. All the corresponding recombinant proteins form various types of covalent oligomers linked by intermolecular disulfide bonds that are reduced in vitro by the thioredoxin (TRX) and/or glutathione/glutaredoxin (GRX) systems. The presence of recombinant MED10a, MED28 and MED32 subunits or changes of its redox state affect the DNA-binding capacity of GeBPL suggesting that redox-driven conformational changes might modulate its activity. Overall, these results advance our understanding of how redox signalling affects transcription and identify mediator as a novel actor in redox signalling pathways, relaying or integrating redox changes in combination with specific TFs as GeBPL. © The Authors Journal compilation © 2015 Biochemical Society.

  2. Characterization of an entomopathogenic fungi target integument protein, Bombyx mori single domain von Willebrand factor type C, in the silkworm, Bombyx mori.

    Science.gov (United States)

    Han, F; Lu, A; Yuan, Y; Huang, W; Beerntsen, B T; Huang, J; Ling, E

    2017-06-01

    The insect cuticle works as the first line of defence to protect insects from pathogenic infections and water evaporation. However, the old cuticle must be shed in order to enter the next developmental stage. During each ecdysis, moulting fluids are produced and secreted into the area among the old and new cuticles. In a previous study, the protein Bombyx mori single domain von Willebrand factor type C (BmSVWC; BGIBMGA011399) was identified in the moulting fluids of Bo. mori and demonstrated to regulate ecdysis. In this study we show that in Bo. mori larvae, BmSVWC primarily locates to the integument (epidermal cells and cuticle), wing discs and head. During the moulting stage, BmSVWC is released into the moulting fluids, and is then produced again by epidermal cells after ecdysis. Fungal infection was shown to decrease the amount of BmSVWC in the cuticle, which indicates that BmSVWC is a target protein of entomopathogenic fungi. Thus, BmSVWC is mainly involved in maintaining the integrity of the integument structure, which serves to protect insects from physical damage and pathogenic infection. © 2017 The Royal Entomological Society.

  3. Characterization of CDOM from urban waters in Northern-Northeastern China using excitation-emission matrix fluorescence and parallel factor analysis.

    Science.gov (United States)

    Zhao, Ying; Song, Kaishan; Li, Sijia; Ma, Jianhang; Wen, Zhidan

    2016-08-01

    Chromophoric dissolved organic matter (CDOM) plays an important role in aquatic systems, but high concentrations of organic materials are considered pollutants. The fluorescent component characteristics of CDOM in urban waters sampled from Northern and Northeastern China were examined by excitation-emission matrix fluorescence and parallel factor analysis (EEM-PARAFAC) to investigate the source and compositional changes of CDOM on both space and pollution levels. One humic-like (C1), one tryptophan-like component (C2), and one tyrosine-like component (C3) were identified by PARAFAC. Mean fluorescence intensities of the three CDOM components varied spatially and by pollution level in cities of Northern and Northeastern China during July-August, 2013 and 2014. Principal components analysis (PCA) was conducted to identify the relative distribution of all water samples. Cluster analysis (CA) was also used to categorize the samples into groups of similar pollution levels within a study area. Strong positive linear relationships were revealed between the CDOM absorption coefficients a(254) (R (2) = 0.89, p CDOM components can be applied to monitor water quality in real time compared to that of traditional approaches. These results demonstrate that EEM-PARAFAC is useful to evaluate the dynamics of CDOM fluorescent components in urban waters from Northern and Northeastern China and this method has potential applications for monitoring urban water quality in different regions with various hydrological conditions and pollution levels.

  4. Functional characterization of secondary wall deposition regulating transcription factors MusaVND2 and MusaVND3 in transgenic banana plants.

    Science.gov (United States)

    Negi, Sanjana; Tak, Himanshu; Ganapathi, T R

    2016-03-01

    NAM, ATAF, and CUC (NAC) domain-containing proteins are plant-specific transcription factors involved in stress responses and developmental regulation. MusaVND2 and MusaVND3 are vascular-related NAC domain-containing genes encoding for nuclear-localized proteins. The transcript level of MusaVND2 and MusaVND3 are gradually induced after induction of lignification conditions in banana embryogenic cells. Banana embryogenic cells differentiated to tracheary element-like cells after overexpression of MusaVND2 and MusaVND3 with a differentiation frequency of 63.5 and 23.4 %, respectively, after ninth day. Transgenic banana plants overexpressing either of MusaVND2 or MusaVND3 showed ectopic secondary wall deposition as well as transdifferentiation of cells into tracheary elements. Transdifferentiation to tracheary element-like cells was observed in cortical cells of corm and in epidermal and mesophyll cells of leaves of transgenic plants. Elevated levels of lignin and crystalline cellulose were detected in the transgenic banana lines than control plants. The results obtained are useful for understanding the molecular regulation of secondary wall development in banana.

  5. Identification of a negative regulatory region for the exchange activity and characterization of T332I mutant of Rho guanine nucleotide exchange factor 10 (ARHGEF10).

    Science.gov (United States)

    Chaya, Taro; Shibata, Satoshi; Tokuhara, Yasunori; Yamaguchi, Wataru; Matsumoto, Hiroshi; Kawahara, Ichiro; Kogo, Mikihiko; Ohoka, Yoshiharu; Inagaki, Shinobu

    2011-08-26

    The T332I mutation in Rho guanine nucleotide exchange factor 10 (ARHGEF10) was previously found in persons with slowed nerve conduction velocities and thin myelination of peripheral nerves. However, the molecular and cellular basis of the T332I mutant is not understood. Here, we show that ARHGEF10 has a negative regulatory region in the N terminus, in which residue 332 is located, and the T332I mutant is constitutively active. An N-terminal truncated ARHGEF10 mutant, ARHGEF10 ΔN (lacking amino acids 1-332), induced cell contraction that was inhibited by a Rho kinase inhibitor Y27632 and had higher GEF activity for RhoA than the wild type. The T332I mutant also showed the phenotype similar to the N-terminal truncated mutant. These data suggest that the ARHGEF10 T332I mutation-associated phenotype observed in the peripheral nerves is due to activated GEF activity of the ARHGEF10 T332I mutant.

  6. Identification of a Negative Regulatory Region for the Exchange Activity and Characterization of T332I Mutant of Rho Guanine Nucleotide Exchange Factor 10 (ARHGEF10)*

    Science.gov (United States)

    Chaya, Taro; Shibata, Satoshi; Tokuhara, Yasunori; Yamaguchi, Wataru; Matsumoto, Hiroshi; Kawahara, Ichiro; Kogo, Mikihiko; Ohoka, Yoshiharu; Inagaki, Shinobu

    2011-01-01

    The T332I mutation in Rho guanine nucleotide exchange factor 10 (ARHGEF10) was previously found in persons with slowed nerve conduction velocities and thin myelination of peripheral nerves. However, the molecular and cellular basis of the T332I mutant is not understood. Here, we show that ARHGEF10 has a negative regulatory region in the N terminus, in which residue 332 is located, and the T332I mutant is constitutively active. An N-terminal truncated ARHGEF10 mutant, ARHGEF10 ΔN (lacking amino acids 1–332), induced cell contraction that was inhibited by a Rho kinase inhibitor Y27632 and had higher GEF activity for RhoA than the wild type. The T332I mutant also showed the phenotype similar to the N-terminal truncated mutant. These data suggest that the ARHGEF10 T332I mutation-associated phenotype observed in the peripheral nerves is due to activated GEF activity of the ARHGEF10 T332I mutant. PMID:21719701

  7. Characterization of the anti-factor VIII immunoglobulin profile in patients with hemophilia A by use of a fluorescence-based immunoassay

    Science.gov (United States)

    Boylan, Brian; Rice, Anne S.; Dunn, Amy L.; Tarantino, Michael D.; Brettler, Doreen B.; Barrett, John C.; Miller, Connie H.

    2015-01-01

    Summary Background The development of neutralizing antibodies, referred to as inhibitors, against factor VIII (FVIII) is a major complication associated with FVIII infusion therapy for the treatment of hemophilia A (HA). Previous studies have shown that a subset of HA patients and a low percentage of healthy individuals harbor non-neutralizing anti-FVIII antibodies that do not elicit the clinical manifestations associated with inhibitor development. Objective Assess HA patients' anti-FVIII antibody profiles as potential predictors of clinical outcomes. Methods A fluorescence immunoassay (FLI) was used to detect anti-FVIII antibodies in 491 samples from 371 HA patients. Results Assessments of antibody profiles showed that the presence of anti-FVIII IgG1, IgG2, or IgG4 correlated qualitatively and quantitatively with the presence of a FVIII inhibitor as reported by the Nijmegen-Bethesda assay (NBA). Forty-eight patients with a negative inhibitor history contributed serial samples to the study, including seven patients who had negative NBA titers initially and later converted to NBA-positive. The FLI detected anti-FVIII IgG1 in five of those seven patients prior to their conversion to NBA-positive. Five of 15 serial-sample patients who had a negative inhibitor history and a positive anti-FVIII IgG1 later developed an inhibitor, compared to 2 of 33 patients with a negative inhibitor history without anti-FVIII IgG1. Conclusions These data provide a rationale for future studies designed both to monitor the dynamics of anti-FVIII antibody profiles in HA patients as a potential predictor of future inhibitor development and to assess the value of the anti-FVIII FLI as a supplement to traditional inhibitor testing. PMID:25354263

  8. Characterization of a Novel Class I Transcription Factor A (CITFA) Subunit That Is Indispensable for Transcription by the Multifunctional RNA Polymerase I of Trypanosoma brucei

    KAUST Repository

    Nguyen, T. N.

    2012-10-26

    Trypanosoma brucei is the only organism known to have evolved a multifunctional RNA polymerase I (pol I) system that is used to express the parasite\\'s ribosomal RNAs, as well as its major cell surface antigens, namely, the variant surface glycoprotein (VSG) and procyclin, which are vital for establishing successful infections in the mammalian host and the tsetse vector, respectively. Thus far, biochemical analyses of the T. brucei RNA pol I transcription machinery have elucidated the subunit structure of the enzyme and identified the class I transcription factor A (CITFA). CITFA binds to RNA pol I promoters, and its CITFA-2 subunit was shown to be absolutely essential for RNA pol I transcription in the parasite. Tandem affinity purification (TAP) of CITFA revealed the subunits CITFA-1 to -6, which are conserved only among kinetoplastid organisms, plus the dynein light chain DYNLL1. Here, by tagging CITFA-6 instead of CITFA-2, a complex was purified that contained all known CITFA subunits, as well as a novel proline-rich protein. Functional studies carried out in vivo and in vitro, as well as a colocalization study, unequivocally demonstrated that this protein is a bona fide CITFA subunit, essential for parasite viability and indispensable for RNA pol I transcription of ribosomal gene units and the active VSG expression site in the mammalian-infective life cycle stage of the parasite. Interestingly, CITFA-7 function appears to be species specific, because expression of an RNA interference (RNAi)-resistant CITFA-7 transgene from Trypanosoma cruzi could not rescue the lethal phenotype of silencing endogenous CITFA-7.

  9. Partial characterization of the cross-reacting determinant, a carbohydrate epitope shared by decay accelerating factor and the variant surface glycoprotein of the African Trypanosoma brucei.

    Science.gov (United States)

    Shak, S; Davitz, M A; Wolinsky, M L; Nussenzweig, V; Turner, M J; Gurnett, A

    1988-03-15

    The variant surface glycoprotein (VSG) of the African trypanosome is anchored in the cell membrane by a complex glycan attached to phosphatidylinositol. The carboxyl terminal portion of VSG contains a cryptic carbohydrate epitope, the cross-reacting determinant (CRD), that is revealed only after removal of the diacylglycerol by phosphatidylinositol-specific phospholipase C (PIPLC) or VSG lipase. Recently, we have shown that after hydrolysis by PIPLC, decay-accelerating factor (DAF)--a mammalian phosphatidylinositol-anchored protein--also contains the CRD epitope. Using a two site immunoradiometric assay in which the capturing antibody is a monoclonal antibody to DAF and the revealing antibody is anti-CRD, we now show that sugar phosphates significantly inhibited the binding of anti-CRD antibody to DAF released by PIPLC. DL-myo-inositol 1,2-cyclic phosphate was the most potent inhibitor of binding (IC50 less than 10(-8) M). Other sugar phosphates, such as alpha-D-glucose-1-phosphate, which also possess adjacent hydroxyl and phosphate moieties in cis also inhibited binding at low concentrations (IC50 = 10(-5) to 10(-4) M). In contrast, sugar phosphates which do not possess adjacent hydroxyl and phosphate moieties in cis and simple sugars weakly inhibited binding (IC50 greater than 10(-3) M). These results suggest that myo-inositol 1,2-cyclic phosphate contributes significantly to the epitope recognized by the anti-CRD antibody and is consistent with analysis of the carboxyl terminus of VSG, which also suggested the presence of the cyclic inositol phosphate. In light of the recent findings that human serum contains a glycan-phosphatidyl-inositol-specific phospholipase D, which converts DAF from a hydrophobic to a hydrophilic form lacking the CRD, the observation that the phosphate is crucial for expression of the epitope may be relevant in understanding the origin of CRD-negative DAF in urine and plasma.

  10. SILAC-MS Based Characterization of LPS and Resveratrol Induced Changes in Adipocyte Proteomics - Resveratrol as Ameliorating Factor on LPS Induced Changes.

    Directory of Open Access Journals (Sweden)

    Mark K Nøhr

    Full Text Available Adipose tissue inflammation is believed to play a pivotal role in the development obesity-related morbidities such as insulin resistance. However, it is not known how this (low-grade inflammatory state develops. It has been proposed that the leakage of lipopolysaccharides (LPS, originating from the gut microbiota, through the gut epithelium could drive initiation of inflammation. To get a better understanding of which proteins and intracellular pathways are affected by LPS in adipocytes, we performed SILAC proteomic analysis and identified proteins that were altered in expression. Furthermore, we tested the anti-inflammatory compound resveratrol. A total of 927 proteins were quantified by the SILAC method and of these 57- and 64 were significantly up- and downregulated by LPS, respectively. Bioinformatic analysis (GO analysis revealed that the upregulated proteins were especially involved in the pathways of respiratory electron transport chain and inflammation. The downregulated proteins were especially involved in protein glycosylation. One of the latter proteins, GALNT2, has previously been described to regulate the expression of liver lipases such as ANGPTL3 and apoC-III affecting lipid metabolism. Furthermore, LPS treatment reduced the protein levels of the insulin sensitizing adipokine, adiponectin, and proteins participating in the final steps of triglyceride- and cholesterol synthesis. Generally, resveratrol opposed the effect induced by LPS and, as such, functioning as an ameliorating factor in disease state. Using an unbiased proteomic approach, we present novel insight of how the proteome is altered in adipocytes in response to LPS as seen in obesity. We suggest that LPS partly exerts its detrimental effects by altering glycosylation processes of the cell, which is starting to emerge as important posttranscriptional regulators of protein expression. Furthermore, resveratrol could be a prime candidate in ameliorating dysfunctioning

  11. Discovery and characterization of a novel lachrymatory factor synthase in Petiveria alliacea and its influence on alliinase-mediated formation of biologically active organosulfur compounds.

    Science.gov (United States)

    Musah, Rabi A; He, Quan; Kubec, Roman

    2009-11-01

    A novel lachrymatory factor synthase (LFS) was isolated and purified from the roots of the Amazonian medicinal plant Petiveria alliacea. The enzyme is a heterotetrameric glycoprotein comprised of two alpha-subunits (68.8 kD each), one gamma-subunit (22.5 kD), and one delta-subunit (11.9 kD). The two alpha-subunits are glycosylated and connected by a disulfide bridge. The LFS has an isoelectric point of 5.2. It catalyzes the formation of a sulfine lachrymator, (Z)-phenylmethanethial S-oxide, only in the presence of P. alliacea alliinase and its natural substrate, S-benzyl-l-cysteine sulfoxide (petiveriin). Depending on its concentration relative to that of P. alliacea alliinase, the LFS sequesters, to varying degrees, the sulfenic acid intermediate formed by alliinase-mediated breakdown of petiveriin. At LFS:alliinase of 5:1, LFS sequesters all of the sulfenic acid formed by alliinase action on petiveriin, and converts it entirely to (Z)-phenylmethanethial S-oxide. However, starting at LFS:alliinase of 5:2, the LFS is unable to sequester all of the sulfenic acid produced by the alliinase, with the result that sulfenic acid that escapes the action of the LFS condenses with loss of water to form S-benzyl phenylmethanethiosulfinate (petivericin). The results show that the LFS and alliinase function in tandem, with the alliinase furnishing the sulfenic acid substrate on which the LFS acts. The results also show that the LFS modulates the formation of biologically active thiosulfinates that are downstream of the alliinase in a manner dependent upon the relative concentrations of the LFS and the alliinase. These observations suggest that manipulation of LFS-to-alliinase ratios in plants displaying this system may provide a means by which to rationally modify organosulfur small molecule profiles to obtain desired flavor and/or odor signatures, or increase the presence of desirable biologically active small molecules.

  12. Functional characterization of a heterologously expressed Brassica napus WRKY41-1 transcription factor in regulating anthocyanin biosynthesis in Arabidopsis thaliana.

    Science.gov (United States)

    Duan, Shaowei; Wang, Jianjun; Gao, Chenhao; Jin, Changyu; Li, Dong; Peng, Danshuai; Du, Guomei; Li, Yiqian; Chen, Mingxun

    2018-03-01

    Previous studies have shown that a plant WRKY transcription factor, WRKY41, has multiple functions, and regulates seed dormancy, hormone signaling pathways, and both biotic and abiotic stress responses. However, it is not known about the roles of AtWRKY41 from the model plant, Arabidopsis thaliana, and its ortholog, BnWRKY41, from the closely related and important oil-producing crop, Brassica napus, in the regulation of anthocyanin biosynthesis. Here, we found that the wrky41 mutation in A. thaliana resulted in a significant increase in anthocyanin levels in rosette leaves, indicating that AtWRKY41 acts as repressor of anthocyanin biosynthesis. RNA sequencing and quantitative real-time PCR analysis revealed increased expression of three regulatory genes AtMYB75, AtMYB111, and AtMYBD, and two structural genes, AT1G68440 and AtGSTF12, all of which contribute to anthocyanin biosynthesis, in the sixth rosette leaves of wrky41-2 plants at 20 days after germination. We cloned the full length complementary DNA of BnWRKY41-1 from the C2 subgenome of the B. napus genotype Westar and observed that, when overexpressed in tobacco leaves as a fusion protein with green fluorescent protein, BnWRKY41-1 is localized to the nucleus. We further showed that overexpression of BnWRKY41-1 in the A. thaliana wrky41-2 mutant rescued the higher anthocyanin content phenotype in rosette leaves of the mutant. Moreover, the elevated expression levels in wrky41-2 rosette leaves of several important regulatory and structural genes regulating anthocyanin biosynthesis were not observed in the BnWRKY41-1 overexpressing lines. These results reveal that BnWRKY41-1 has a similar role with AtWRKY41 in regulating anthocyanin biosynthesis when overexpressed in A. thaliana. This gene represents a promising target for genetically manipulating B. napus to increase the amounts of anthocyanins in rosette leaves. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. Seasonal characterization of CDOM for lakes in semi-arid regions of Northeast China using excitation-emission matrices fluorescence and parallel factor analysis (EEM-PARAFAC)

    Science.gov (United States)

    Zhao, Y.; Song, K.; Wen, Z.; Li, L.; Zang, S.; Shao, T.; Li, S.; Du, J.

    2015-04-01

    The seasonal characteristics of fluorescence components in CDOM for lakes in the semi-arid region of Northeast China were examined by excitation-emission matrices fluorescence and parallel factor analysis (EEM-PARAFAC). Two humic-like peaks C1 (Ex/Em = 230, 300/425 nm) and C2 (Ex/Em = 255, 350/460 nm) and two protein-like B (Ex/Em = 220, 275/320 nm) and T (Ex/Em = 225, 290/360 nm) peaks were identified using PARAFAC. The average fluorescence intensity of the four components differed with seasonal variation from June and August 2013 to February and April 2014. The total fluorescence intensity significantly varied from 2.54 ± 0.68 nm-1 in June to the mean value 1.93 ± 0.70 nm-1 in August 2013, and then increased to 2.34 ± 0.92 nm-1 in February and reduced to the lowest 1.57 ± 0.55 nm-1 in April 2014. In general, the fluorescence intensity was dominated by peak C1, indicating that most part of CDOM for inland waters being investigated in this study was originated from phytoplankton degradation. The lowest C2 represents only a small portion of CDOM from terrestrial imported organic matter to water bodies through rainwash and soil leaching. The two protein-like intensities (B and T) formed in situ through microbial activity have almost the same intensity. Especially, in August 2013 and February 2014, the two protein-like peaks showed obviously difference from other seasons and the highest C1 (1.02 nm-1) was present in February 2014. Components 1 and 2 exhibited strong linear correlation (R2 = 0.633). There were significantly positive linear relationships between CDOM absorption coefficients a(254) (R2 = 0.72, 0.46, p DOC. However, almost no obvious correlation was found between salinity and EEM-PARAFAC extracted components except for C3 (R2 = 0.469). Results from this investigation demonstrate that the EEM-PARAFAC technique can be used to evaluate the seasonal dynamics of CDOM fluorescence components for inland waters in semi-arid regions of Northeast China.

  14. [Characterizing chromophoric dissolved organic matter (CDOM) in Lake Honghu, Lake Donghu and Lake Liangzihu using excitation-emission matrices (EEMs) fluorescence and parallel factor analysis (PARAFAC)].

    Science.gov (United States)

    Zhou, Yong-Qiang; Zhang, Yun-Lin; Niu, Cheng; Wang, Ming-Zhu

    2013-12-01

    Little is known about DOM characteristics in medium to large sized lakes located in the middle and lower reaches of Yangtze River, like Lake Honghu, Lake Donghu and Lake Liangzihu. Absorption, fluorescence and composition characteristics of chromophoric dissolved organic matter (CDOM) are presented using the absorption spectroscopy, the excitation-emission ma trices (EEMs) fluorescence and parallel factor analysis (PARAFAC) model based on the data collected in Sep-Oct. 2007 including 15, 9 and 10 samplings in Lake Honghu, Lake Donghu and Lake Liangzihu, respectively. CDOM absorption coefficient at 350 nm a(350) coefficient in Lake Honghu was significantly higher than those in Lake Donghu and Lake Liangzihu (t-test, pCDOM spectral slope in the wavelength range of 280-500 nm (S280-500) and a(350) (R2 =0. 781, p<0. 001). The mean value of S280-500 in Lake Honghu was significantly lower than those in Lake Donghu (t-test, p

  15. Characterization of insulin-like growth factor I receptors in the median eminence of the brain and their modulation by food restriction

    International Nuclear Information System (INIS)

    Bohannon, N.J.; Corp, E.S.; Wilcox, B.J.; Figlewicz, D.P.; Dorsa, D.M.; Baskin, D.G.

    1988-01-01

    High affinity binding sites for 125I-labeled [Thr59]insulin-like growth factor I (IGF-I) were measured in rat median eminence by in vitro autoradiography with slide-mounted sections of frozen rat brain. Specific binding of 0.1 nM iodo-[Thr59]IGF-I to brain slices reached maximum by 12 h at 4 C and was unchanged at 24 h. Densitometry by computer digital image analysis of autoradiographic images indicated that specific binding of iodo-[Thr59]IGF-I to the median eminence was reversible. The specificity of binding was evaluated with competition of iodo-[Thr59]IGF-I with unlabeled [Thr59]IGF-I, rat IGF-II (multiplication-stimulating activity), and porcine insulin. All were recognized by the binding site, but the rank order of potency was [Thr59]IGF-I greater than IGF-II greater than insulin. Somatostatin was completely ineffective. Further, an antibody against the rat IGF-II receptor did not block binding of iodo-[Thr59]IGF-I to the median eminence. Fourteen days of food restriction (75% of food intake of controls) resulted in significant weight loss and reduction of plasma immunoreactive IGF-I in six food-restricted rats (0.9 +/- 0.1 U/ml) compared with values in six controls (2.6 +/- 0.5 U/ml; P less than 0.001). Binding of 125I-labeled [Thr59]IGF-I in the median eminence was significantly increased in the food-restricted rats, primarily due to an increase in the concentration of iodo-[Thr59]IGF-I-binding sites in the median eminence; the affinity (Kd) of binding was unchanged. The results indicate that the median eminence has type I IGF-I receptors, which become more numerous under metabolic conditions associated with decreased caloric intake and lowered plasma IGF-I levels

  16. Microbiological water quality of Igapó Lake Londrina - PR and genotypic characterization of virulence factors associated with enteropathogenic Escherichia coli (EPEC and Shiga toxin-producing E. coli (STEC

    Directory of Open Access Journals (Sweden)

    Paulo Alfonso Schuroff

    2014-12-01

    Full Text Available This work aimed at the detection and quantification of Total Coliforms and Escherichia coli in Igapó Lake, in order to evaluate the quality of these waters as proper or unfit for recreation, in addition to the genotypic characterization of virulence factors associated with EPEC and STEC by PCR. The study area was the Igapó Lake I, II, III and IV. Samples were collected monthly from March 2011 to February 2012. The technique used for the detection and quantification of Total Coliforms and E. coli was the Colilert chromogenic substrate. In the technique of PCR, the eae and bfp genes were tested to characterize the typical EPEC pathotype; stx1, stx2, eae and hlyA the STEC pathotype and the samples that presented only the eae gene were characterized as atypical EPEC. According to CONAMA Resolution 357/2005, it has been observed that only Igapó Lake III was rated inappropriate for primary contact recreation, while for secondary contact recreation, all lakes were considered appropriate. Moreover, a strong relationship between rainfall and E. coli indices in Igapó Lake can be observed, which in dry months the quantity drastically decreases, while in rainy months the opposite relationship was observed. Of the 97 strains of E. coli isolated, two had the eae gene (atypical EPEC. None of the isolates contained genes stx1, stx2, bfp and hlyA. Thus, we hope to educate the population and public agencies of the importance of microbiological monitoring of recreational waters to prevent outbreaks of waterborne infections.

  17. Inventive activity of the Department of Protein Structure and Function of the Palladin Institute of Biochemistry of NAS of Ukraine. Part I. Development of the diagnostic methods for detection of hemostasis disorders and characterization of certain blood coagulation factors

    Directory of Open Access Journals (Sweden)

    V. M. Danilova

    2016-04-01

    Full Text Available The practical aspects of inventive activity of the Department of Protein Structure and Function of the Palladin Institute of Biochemistry, NAS of Ukraine are highlighted in this article. Through years of fundamental and applied researches of blood coagulation system proteins, initiated by luminaries of the world biochemistry O. V. Palladin and V. O. Belitser, the Department staff have developed a considerable number of methods, techniques and tests for the assessment of the state of the hemostasis system, which were approved in many clinics. In the first part of this work the authors describe the development of the diagnostic methods for identifying the homeostasis system disorders in detail, as well as characterize certain coagulation factors.

  18. [Characterization of Chromophoric dissolved organic matter (CDOM) in Zhoushan fishery using excitation-emission matrix spectroscopy (EEMs) and parallel factor analysis (PARAFAC)].

    Science.gov (United States)

    Zhou, Qian-qian; Su, Rong-guo; Bai, Ying; Zhang, Chuan-song; Shi, Xiao-yong

    2015-01-01

    The composition, distribution characteristics and sources of chromophoric dissolved organic matter(CDOM) in Zhoushan Fishery in spring were evaluated by fluorescence excitation-emission matrix (EEM) combined with parallel factor analysis (EEMs-PARAFAC). Three humic-like components [C1 (330/420 nm)], C2 [(290) 365/440 nm] and C3 [(260) 370/490 nm)] and two protein-like components [C4(285/340 nm) and C5 (270/310 nm)] were identified by EEMs-PARAFAC. The horizontal distribution patterns of the five components were almost the same with only slight differences, showing decreasing trends with increasing distance from shore. In the surface and middle layers, the high value areas were located in the north of Hangzhou Bay estuary and the outlet of Xiazhimen channel, and the former's was higher in the surface layer while the latter's was higher in the middle layer. In the bottom layer, CDOM decreased gradiently from the inshore to offshore, with higher CDOM near Zhoushan Island. The distributions of fluorescence components showed an opposite trend with salinity, and no significant linear relationship with Chl-a concentration was found, which indicated that CDOM in the surface and middle layers were dominated by terrestrial input and human activities of Zhoushan Island and that of the bottom layer was attribute to human activities of Zhoushan Island. The vertical distribution of five fluorescent components along 30.5 degrees N transect showed a decreasing trend from the surface and middle layers to bottom layer with high values in inshore and offshore areas, which were correlated with the lower salinity and higher Chl-a concentration, respectively. On this transect, CDOM was mainly affected by Yangtze River input in coastal area but by bioactivities in offshore waters. Along the 30 degrees N transect, the vertical distribution patterns of CDOM were similar to those of 30.5 degrees N transect but there was a high value area in the bottom layer near the shore, attributing to

  19. Diabetes and Obesity as Independent Risk Factors for Osteoporosis: Updated Results from the ROIS/EMEROS Registry in a Population of Five Thousand Post-Menopausal Women Living in a Region Characterized by Heavy Environmental Pressure.

    Science.gov (United States)

    Neglia, Cosimo; Argentiero, Alberto; Chitano, Giovanna; Agnello, Nadia; Ciccarese, Roberta; Vigilanza, Antonella; Pantile, Valerio; Argentiero, Domenico; Quarta, Raffaele; Rivezzi, Matteo; Di Tanna, Gian Luca; Di Somma, Carolina; Migliore, Alberto; Iolascon, Giovanni; Gimigliano, Francesca; Distante, Alessandro; Piscitelli, Prisco

    2016-11-01

    Objectives : We aimed to analyze bone mineralization and the effect of different risk factors for osteoporosis in postmenopausal women. Methods : We found 4909 postmenopausal subjects within ≥10,000 records from the ROIS/EMEROS (Ionian and Salento Osteoporosis Registry/Euro Mediterranean Registry of Osteoporosis) registry, a population study carried out in an area characterized by heavy environmental pressure between Brindisi and Taranto from 2009 to 2016. All subjects were assessed via phalangeal quantitative ultrasound (QUS) to evaluate their bone mineralization (assessed via amplitude dependent speed of sound (AD-SoS)) and the association between demineralization and the presence of other conditions or risk factors. Results : Mean age was 64 ± 9.5 years and mean body mass index (BMI) was 28.7 ± 3.5 kg/m². Pearson correlation analyses revealed a negative association between bone mineralization (AD-SoS) and BMI ( p osteoporosis (adjusted for age, physical activity, and the use of drugs known to increase the risk of fractures) in subjects with diabetes and obesity: 1.39 (confidence interval (CI): 1.05-1.83) and 1.46 (CI: 1.20-1.78), respectively. A statistically significant linear trend of higher ORs of osteoporosis was found for increasing values of BMI. Conclusions : Our study confirmed the high impact of obesity and type 1 and type 2 diabetes on osteoporosis.

  20. The primary extinction and static Debye-Waller factor in the characterization of textured nickel by X-ray diffraction; La extincion primaria y el factor estatico de Debye-Waller en la caracterizacion de niquel con textura mediante difraccion de rayos X

    Energy Technology Data Exchange (ETDEWEB)

    Kryshtab, T.; Palacios G, J. [Instituto Politecnico Nacional, Escuela Superior de Fisica y Matematicas, Av. IPN s/n, 07738 Mexico D. F. (Mexico); Cadena A, A. [Instituto Politecnico Nacional, Escuela Superior de Ingenieria Quimica e Industrias Extractivas, Av. IPN s/n, 07738 Mexico D. F. (Mexico); Kryvko, A., E-mail: kryshtab@gmail.com [Instituto Politecnico Nacional, Escuela Superior de Ingenieria Mecanica y Electrica, Unidad Zacatenco, 07360 Mexico D. F. (Mexico)

    2015-07-01

    The texture analysis using X-ray diffraction (XRD) implies measurement of pole figures (Pf) from the diffracted intensities considering the model of kinematical dispersion. The extinction phenomenon results in a decrease of diffracted intensity and that in turn in a decrease of pole densities (Pds). The phenomenon appears in the kinematical theory of XRD as the primary extinction and the secondary extinction to characterize the loss of intensity of kinematical dispersion. In turn, the static Debye-Waller factor is an integral characteristic of defects in crystals that is introduced in the kinematical theory of XRD and also is used in dynamical theory of XRD. In this work the correlation between the primary extinction coefficient and the static Debye-Waller factor in the case of textured nickel was determined. The value of static Debye-Waller factor was determined from the value of the calculated primary extinction coefficient. For the evaluation there were used Pds in the maxima of Pf obtained for 111 and 200 reflections with Mo Kα radiation, and the Pds in the maxima of Pf obtained for the first and second orders of these reflections with Cu Kα and Co Kα radiations. There were calculated the dislocation densities in grains using values of static Debye-Waller factor and the extinction coefficients. The dislocation densities calculated from these two characteristics are practically equal. (Author)

  1. Characterization of StABF1, a stress-responsive bZIP transcription factor from Solanum tuberosum L. that is phosphorylated by StCDPK2 in vitro.

    Science.gov (United States)

    Muñiz García, María Noelia; Giammaria, Verónica; Grandellis, Carolina; Téllez-Iñón, María Teresa; Ulloa, Rita María; Capiati, Daniela Andrea

    2012-04-01

    ABF/AREB bZIP transcription factors mediate plant abiotic stress responses by regulating the expression of stress-related genes. These proteins bind to the abscisic acid (ABA)-responsive element (ABRE), which is the major cis-acting regulatory sequence in ABA-dependent gene expression. In an effort to understand the molecular mechanisms of abiotic stress resistance in cultivated potato (Solanum tuberosum L.), we have cloned and characterized an ABF/AREB-like transcription factor from potato, named StABF1. The predicted protein shares 45-57% identity with A. thaliana ABFs proteins and 96% identity with the S. lycopersicum SlAREB1 and presents all of the distinctive features of ABF/AREB transcription factors. Furthermore, StABF1 is able to bind to the ABRE in vitro. StABF1 gene is induced in response to ABA, drought, salt stress and cold, suggesting that it might be a key regulator of ABA-dependent stress signaling pathways in cultivated potato. StABF1 is phosphorylated in response to ABA and salt stress in a calcium-dependent manner, and we have identified a potato CDPK isoform (StCDPK2) that phosphorylates StABF1 in vitro. Interestingly, StABF1 expression is increased during tuber development and by tuber-inducing conditions (high sucrose/nitrogen ratio) in leaves. We also found that StABF1 calcium-dependent phosphorylation is stimulated by tuber-inducing conditions and inhibited by gibberellic acid, which inhibits tuberization.

  2. Mass spectrometric characterization of the hypoxia-inducible factor (HIF) stabilizer drug candidate BAY 85-3934 (molidustat) and its glucuronidated metabolite BAY-348, and their implementation into routine doping controls.

    Science.gov (United States)

    Dib, Josef; Mongongu, Cynthia; Buisson, Corinne; Molina, Adeline; Schänzer, Wilhelm; Thuss, Uwe; Thevis, Mario

    2017-01-01

    The development of new therapeutics potentially exhibiting performance-enhancing properties implicates the risk of their misuse by athletes in amateur and elite sports. Such drugs necessitate preventive anti-doping research for consideration in sports drug testing programmes. Hypoxia-inducible factor (HIF) stabilizers represent an emerging class of therapeutics that allows for increasing erythropoiesis in patients. BAY 85-3934 is a novel HIF stabilizer, which is currently undergoing phase-2 clinical trials. Consequently, the comprehensive characterization of BAY 85-3934 and human urinary metabolites as well as the implementation of these analytes into routine doping controls is of great importance. The mass spectrometric behaviour of the HIF stabilizer drug candidate BAY 85-3934 and a glucuronidated metabolite (BAY-348) were characterized by electrospray ionization-(tandem) mass spectrometry (ESI-MS(/MS)) and multiple-stage mass spectrometry (MS n ). Subsequently, two different laboratories established different analytical approaches (one each) enabling urine sample analyses by employing either direct urine injection or solid-phase extraction. The methods were cross-validated for the metabolite BAY-348 that is expected to represent an appropriate target analyte for human urine analysis. Two test methods allowing for the detection of BAY-348 in human urine were applied and cross-validated concerning the validation parameters specificity, linearity, lower limit of detection (LLOD; 1-5 ng/mL), ion suppression/enhancement (up to 78%), intra- and inter-day precision (3-21%), recovery (29-48%), and carryover. By means of ten spiked test urine samples sent blinded to one of the participating laboratories, the fitness-for-purpose of both assays was provided as all specimens were correctly identified applying both testing methods. As no post-administration study samples were available, analyses of authentic urine specimens remain desirable. Copyright © 2016 John Wiley

  3. Diabetes and Obesity as Independent Risk Factors for Osteoporosis: Updated Results from the ROIS/EMEROS Registry in a Population of Five Thousand Post-Menopausal Women Living in a Region Characterized by Heavy Environmental Pressure

    Directory of Open Access Journals (Sweden)

    Cosimo Neglia

    2016-11-01

    Full Text Available Objectives: We aimed to analyze bone mineralization and the effect of different risk factors for osteoporosis in postmenopausal women. Methods: We found 4909 postmenopausal subjects within ≥10,000 records from the ROIS/EMEROS (Ionian and Salento Osteoporosis Registry/Euro Mediterranean Registry of Osteoporosis registry, a population study carried out in an area characterized by heavy environmental pressure between Brindisi and Taranto from 2009 to 2016. All subjects were assessed via phalangeal quantitative ultrasound (QUS to evaluate their bone mineralization (assessed via amplitude dependent speed of sound (AD-SoS and the association between demineralization and the presence of other conditions or risk factors. Results: Mean age was 64 ± 9.5 years and mean body mass index (BMI was 28.7 ± 3.5 kg/m2. Pearson correlation analyses revealed a negative association between bone mineralization (AD-SoS and BMI (p < 0.001. By using multivariate logistic regression analysis, we observed significant values of odds ratios (ORs of osteoporosis (adjusted for age, physical activity, and the use of drugs known to increase the risk of fractures in subjects with diabetes and obesity: 1.39 (confidence interval (CI: 1.05–1.83 and 1.46 (CI: 1.20–1.78, respectively. A statistically significant linear trend of higher ORs of osteoporosis was found for increasing values of BMI. Conclusions: Our study confirmed the high impact of obesity and type 1 and type 2 diabetes on osteoporosis.

  4. Characterization of insulin-like-growth factor II (IGF II mRNA positive hepatic altered foci and IGF II expression in hepatocellular carcinoma during diethylnitrosamine-induced hepatocarcinogenesis in rats

    Directory of Open Access Journals (Sweden)

    Mukherjee Biswajit

    2005-08-01

    Full Text Available Abstract Background Insulin-like-growth factor II (IGF II has been implicated in the pathogenesis of neoplasm of different tissues, including liver of rats and men. This growth factor is believed to exert its effect during cellular proliferation. During the process of development of hepatocellular carcinoma (HCC, different hepatic altered foci appear. They are believed to be the putative precursors of HCC in rats and in men. Thus, to study the role of the gene in a defined model of hepatocarcinogenesis was the target to elucidate its role in various cancer phenotypes during the entire development stage of cancer, right from earlier preneoplastic lesions to HCC Methods Antisense in situ hybridization technique was used here to characterize the type(s of foci in which IGF II mRNA had expressed during the development of hepatocarcinogenesis-induced by diethylnitrosamine and promoted by phenobarbital in rats. Various focal lesions have been categorized depending on the stages and sizes along with IGF II expression patterns in them. Immunohistochemical detection for proliferating cell nuclear antigen (PCNA was made to detect the role of the gene in preneoplastic and neoplastic cellular proliferation. Results IGF II expression was located in the glycogen-storage acidophilic cell foci maximally followed by mixed cell lesions and the least in basophilic lesions. The expression of IGF II was found to be predominant in the HCC. The expression of gene was also located at the peripheral cells of spongiosis hepatis which are believed to be the precursor of ito cell carcinoma. It was noted that there is a direct correlation between IGF II expression and Immunohistochemical detection for PCNA. Conclusion It may be concluded that IGF II gene expression plays an important role during the development of neoplasia and the gene expresses in the sequence of events leading from glycogen-rich-acidophilic lesions to glycogen poor basophilic lesions to HCC with an

  5. Characterizing multifragmentation

    International Nuclear Information System (INIS)

    Campi, X.; Krivine, H.

    1994-01-01

    Various methods to characterize the fragment size distributions in nuclear multifragmentation are discussed. The goal is to find the best signals of a phase transition associated to multifragmentation. The concepts of scaling and critical exponents are reviewed and the possibility to determine them in finite nuclei is examined. The fluctuations of the fragment size distribution and a possible signal of intermittency are also discussed. (author). 29 refs., 4 figs., 1 tab

  6. Hybrid Capture-Based Comprehensive Genomic Profiling Identifies Lung Cancer Patients with Well-Characterized Sensitizing Epidermal Growth Factor Receptor Point Mutations That Were Not Detected by Standard of Care Testing.

    Science.gov (United States)

    Suh, James H; Schrock, Alexa B; Johnson, Adrienne; Lipson, Doron; Gay, Laurie M; Ramkissoon, Shakti; Vergilio, Jo-Anne; Elvin, Julia A; Shakir, Abdur; Ruehlman, Peter; Reckamp, Karen L; Ou, Sai-Hong Ignatius; Ross, Jeffrey S; Stephens, Philip J; Miller, Vincent A; Ali, Siraj M

    2018-03-14

    In our recent study, of cases positive for epidermal growth factor receptor ( EGFR ) exon 19 deletions using comprehensive genomic profiling (CGP), 17/77 (22%) patients with prior standard of care (SOC) EGFR testing results available were previously negative for exon 19 deletion. Our aim was to compare the detection rates of CGP versus SOC testing for well-characterized sensitizing EGFR point mutations (pm) in our 6,832-patient cohort. DNA was extracted from 40 microns of formalin-fixed paraffin-embedded sections from 6,832 consecutive cases of non-small cell lung cancer (NSCLC) of various histologies (2012-2015). CGP was performed using a hybrid capture, adaptor ligation-based next-generation sequencing assay to a mean coverage depth of 576×. Genomic alterations (pm, small indels, copy number changes and rearrangements) involving EGFR were recorded for each case and compared with prior testing results if available. Overall, there were 482 instances of EGFR exon 21 L858R (359) and L861Q (20), exon 18 G719X (73) and exon 20 S768I (30) pm, of which 103 unique cases had prior EGFR testing results that were available for review. Of these 103 cases, CGP identified 22 patients (21%) with sensitizing EGFR pm that were not detected by SOC testing, including 9/75 (12%) patients with L858R, 4/7 (57%) patients with L861Q, 8/20 (40%) patients with G719X, and 4/7 (57%) patients with S768I pm (some patients had multiple EGFR pm). In cases with available clinical data, benefit from small molecule inhibitor therapy was observed. CGP, even when applied to low tumor purity clinical-grade specimens, can detect well-known EGFR pm in NSCLC patients that would otherwise not be detected by SOC testing. Taken together with EGFR exon 19 deletions, over 20% of patients who are positive for EGFR -activating mutations using CGP are previously negative by SOC EGFR mutation testing, suggesting that thousands of such patients per year in the U.S. alone could experience improved clinical

  7. Hyperglycemia: a prothrombotic factor?

    NARCIS (Netherlands)

    Lemkes, B. A.; Hermanides, J.; DeVries, J. H.; Holleman, F.; Meijers, J. C. M.; Hoekstra, J. B. L.

    2010-01-01

    Diabetes mellitus is characterized by a high risk of atherothrombotic events. What is more, venous thrombosis has also been found to occur more frequently in this patient group. This prothrombotic condition in diabetes is underpinned by laboratory findings of elevated coagulation factors and

  8. Genotypic characterization of virulence factors in Escherichia coli strains from patients with cystitis Caracterização genotípica dos fatores de virulência em amostras de Escherichia coli isoladas de pacientes com cistite

    Directory of Open Access Journals (Sweden)

    Monique Ribeiro Tiba

    2008-10-01

    Full Text Available Adhesins (P-fimbriae, S-fimbriae, type 1 fimbriae and afimbrial adhesin, toxins (α-hemolysin and cytotoxic necrotizing factor type 1, iron acquisition systems (aerobactin and host defense avoidance mechanisms (capsule or lipopolysaccharide have been shown to be prevalent in Escherichia coli strains associated with urinary tract infections. In this work, 162 Uropathogenic Escherichia coli (UPEC strains from patients with cystitis were genotypically characterized by polymerase chain reaction (PCR assay. We developed three multiplex PCR assays for virulence-related genes papC, papE/F, papG alleles, fimH, sfa/foc, afaE, hly, cnf-1, usp, cdtB, iucD, and kpsMTII, all of them previously identified in UPEC strains. The PCR assay results identified 158 fimH (97.5%, 86 kpsMTII (53.1%, 53 papC/papEF/papG (32.7%, 45 sfa (27.8%, 42 iucD (25.9%, 41 hly (25.3%, 36 usp (22.2%, 30 cnf-1(18.5% and 10 afa (6.2% strains. No strain was positive for cdtB. In this work, we also demonstrated that adhesins may be multiple within a single strain and that several virulence genes can occur combined in association.Adesinas (Fímbria P, fímbria S, fímbria do tipo 1 e a adesina afimbrial, toxinas (α-hemolisina e o fator necrosante citotóxico do tipo 1, sistemas de captação de ferro (aerobactina, e mecanismos de defesa do hospedeiro (cápsula ou lipopolissacarídeo são prevalentes em amostras de Escherichia coli associadas a infecções do trato urinário. O objetivo deste trabalho foi caracterizar genotipicamente 162 amostras de Escherichia coli uropatogênica (UPEC de pacientes com cistite através do ensaio da reação em cadeia da polimerase. Foram realizados três ensaios de PCR multiplex para os seguintes fatores de virulência: papC, papE/F, alelos de papG, fimH, sfa/foc, afaE, hly, cnf-1, usp, cdtB, iucD, e kpsMTII. Os resultados da PCR identificaram, 158 amostras fimH (97,5%, 86 amostras kpsMTII (53,1%, 53 amostras papC/papEF/papG (32,7%, 45 amostras sfa (27

  9. Risk Factors

    Science.gov (United States)

    ... cells do not invade nearby tissues or spread. Risk Factors Key Points Factors That are Known to ... chemicals . Factors That are Known to Increase the Risk of Cancer Cigarette Smoking and Tobacco Use Tobacco ...

  10. CCD characterization and measurements automation

    Czech Academy of Sciences Publication Activity Database

    Kotov, I.V.; Frank, J.; Kotov, A.I.; Kubánek, Petr; O´Connor, P.; Prouza, Michael; Radeka, V.; Takacs, P.

    2012-01-01

    Roč. 695, Dec (2012), 188-192 ISSN 0168-9002 R&D Projects: GA MŠk ME09052 Institutional research plan: CEZ:AV0Z10100502 Keywords : CCD * characterization * test automation Subject RIV: BN - Astronomy, Celestial Mechanics, Astrophysics Impact factor: 1.142, year: 2012

  11. Characterization of a thrombin cleavage site mutation (Arg 1689 to Cys) in the factor VIII gene of two unrelated patients with cross-reacting material-positive hemophilia A.

    Science.gov (United States)

    Arai, M; Higuchi, M; Antonarakis, S E; Kazazian, H H; Phillips, J A; Janco, R L; Hoyer, L W

    1990-01-15

    The molecular defect responsible for moderate and severe hemophilia A has been identified for two unrelated patients with the CRM-positive form of this disorder (factor VIII activity of 0.02 and 0.05 U/mL with factor VIII antigen of 0.87 and 2.20 U/mL). In both cases, the immunopurified dysfunctional factor VIII protein is abnormal, in that the 80 Kd light chain is not cleaved by thrombin at arginine-1689. The basis for this failure was identified by polymerase chain reaction amplification of exon 14 of the variant factor VIII genes and direct sequencing of the amplified products. In both cases, a single base substitution (C to T) was identified that produces an arginine to cysteine substitution at amino acid residue 1689. These data identify the molecular defects of the two identical factor VIII variant proteins. The dysfunctional factor VIII has been designated "Factor VIII-East Hartford," the residence of the patient in whom the defect was first identified.

  12. Biomass Characterization | Bioenergy | NREL

    Science.gov (United States)

    Characterization Biomass Characterization NREL provides high-quality analytical characterization of biomass feedstocks, intermediates, and products, a critical step in optimizing biomass conversion clear, amber liquid Standard Biomass Laboratory Analytical Procedures We maintain a library of

  13. Functional Characterization of the Role of the N-terminal Domain of the c/Nip1 Subunit of Eukaryotic Initiation Factor 3 (eIF3) in AUG Recognition

    Czech Academy of Sciences Publication Activity Database

    Karásková, Martina; Gunišová, Stanislava; Herrmannová, Anna; Wagner, Susan; Munzarová, Vanda; Valášek, Leoš Shivaya

    2012-01-01

    Roč. 287, č. 34 (2012), s. 28420-28434 ISSN 0021-9258 R&D Projects: GA ČR GAP305/10/0335 Institutional support: RVO:61388971 Keywords : START CODON SELECTION * 40S RIBOSOMAL-SUBUNIT * GCN4 TRANSLATIONAL CONTROL Subject RIV: CE - Biochemistry Impact factor: 4.651, year: 2012

  14. Nanotechnology Characterization Lab

    Data.gov (United States)

    Federal Laboratory Consortium — The Nanotechnology Characterization Laboratory (NCL) at the Frederick National Laboratory for Cancer Research performs preclinical characterization of nanomaterials...

  15. Materials characterization techniques

    National Research Council Canada - National Science Library

    Zhang, Sam; Li, L; Kumar, Ashok

    2009-01-01

    "With an emphasis on practical applications and real-world case studies, Materials Characterization Techniques presents the principles of widely used advanced surface and structural characterization...

  16. Nanotechnology Characterization Laboratory

    Data.gov (United States)

    Federal Laboratory Consortium — The Nanotechnology Characterization Laboratory (NCL) at the Frederick National Laboratory for Cancer Research performs preclinical characterization of nanomaterials...

  17. Factor XII (Hageman factor) deficiency

    Science.gov (United States)

    ... this page: //medlineplus.gov/ency/article/000545.htm Factor XII (Hageman factor) deficiency To use the sharing features on this ... M. is also a founding member of Hi-Ethics and subscribes to the principles of the Health ...

  18. Emission factors and congener-specific characterization of PCDD/Fs, PCBs, PBDD/Fs and PBDEs from an off-road diesel engine using waste cooking oil-based biodiesel blends.

    Science.gov (United States)

    Chen, Shui-Jen; Tsai, Jen-Hsiung; Chang-Chien, Guo-Ping; Huang, Kuo-Lin; Wang, Lin-Chi; Lin, Wen-Yinn; Lin, Chih-Chung; Yeh, C Kuei-Jyum

    2017-10-05

    Few studies have been performed up to now on the emission factors and congener profiles of persistent organic pollutants (POPs) emitted from off-road diesel engines. This investigation elucidates the emission factors and congener profiles of various POPs, namely polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs), polychlorinated biphenyl (PCBs), polybrominated dibenzo-p-dioxins and polybrominated dibenzofurans (PBDD/Fs) and polybrominated diphenyl ethers (PBDEs), in the exhausts of a diesel generator fueled with different waste cooking oil-based biodiesel (WCO-based biodiesel) blends. The PCDD/Fs contributed 87.2% of total dioxin-like toxicity (PCDD/Fs+PCBs+PBDD/Fs) in the exhaust, while the PCBs and PBDD/Fs only contributed 8.2% and 4.6%, respectively. Compared with petroleum diesel, B20 (20vol% WCO-based biodiesel+80vol% diesel) reduced total toxicity by 46.5% for PCDD/Fs, 47.1% for PCBs, and 24.5% for PBDD/Fs, while B40 (40vol% WCO-based biodiesel+60vol% diesel) reduced it by 89.5% for PCDD/Fs, 57.1% for PCBs, and 63.2% for PBDD/Fs in POP emission factors. The use of WCO-based biodiesel not only solves the problem of waste oil disposal, but also lowers POP emissions from diesel generators. Copyright © 2017 Elsevier B.V. All rights reserved.

  19. Molecular characterization and mRNA expression of two key enzymes of hypoxia-sensing pathways in eastern oysters Crassostrea virginica (Gmelin): Hypoxia-inducible factor α (HIF-α) and HIF-prolyl hydroxylase (PHD)

    Science.gov (United States)

    Piontkivska, Helen; Chung, J. Sook; Ivanina, Anna V.; Sokolov, Eugene P.; Techa, Sirinart; Sokolova, Inna M.

    2010-01-01

    Oxygen homeostasis is crucial for development, survival and normal function of all metazoans. A family of transcription factors called hypoxia-inducible factors (HIF) is critical in mediating the adaptive responses to reduced oxygen availability. The HIF transcription factor consists of a constitutively expressed β subunit and an oxygen-dependent α subunit; the abundance of the latter determines the activity of HIF and is regulated by a family of O2- and Fe2+-dependent enzymes prolyl hydroxylases (PHDs). Currently very little is known about the function of this important pathway and the molecular structure of its key players in hypoxia-tolerant intertidal mollusks including oysters, which are among the animal champions of anoxic and hypoxic tolerance and thus can serve as excellent models to study the role of HIF cascade in adaptations to oxygen deficiency. We have isolated transcripts of two key components of the oxygen sensing pathway - the oxygen-regulated HIF-α subunit and PHD - from an intertidal mollusk, the eastern oyster Crassostrea virginica, and determined the transcriptional responses of these two genes to anoxia, hypoxia and cadmium (Cd) stress. HIF-α and PHD homologs from eastern oysters C. virginica show significant sequence similarity and share key functional domains with the earlier described isoforms from vertebrates and invertebrates. Phylogenetic analysis shows that genetic diversification of HIF and PHD isoforms occurred within the vertebrate lineage indicating functional diversification and specialization of the oxygen-sensing pathways in this group, which parallels situation observed for many other important genes. HIF-α and PHD homologs are broadly expressed at the mRNA level in different oyster tissues and show transcriptional responses to prolonged hypoxia in the gills consistent with their putative role in oxygen sensing and the adaptive response to hypoxia. Similarity in amino acid sequence, domain structure and transcriptional

  20. Robust factorization

    DEFF Research Database (Denmark)

    Aanæs, Henrik; Fisker, Rune; Åström, Kalle

    2002-01-01

    Factorization algorithms for recovering structure and motion from an image stream have many advantages, but they usually require a set of well-tracked features. Such a set is in generally not available in practical applications. There is thus a need for making factorization algorithms deal effect...

  1. Organizational factors

    International Nuclear Information System (INIS)

    Holy, J.

    1999-12-01

    The following organizational factors are considered with respect to the human factor and operating safety of nuclear power plants: external influences; objectives and strategy; positions and ways of management; allocation of resources; working with human resources; operators' training; coordination of work; knowledge of organization and management; proceduralization of the topic; labour organizing culture; self-improvement system; and communication. (P.A.)

  2. El factoring

    Directory of Open Access Journals (Sweden)

    Alberto Rosenthal

    1988-04-01

    Full Text Available RESUMEN El artículo  presenta, una conceptualización general de lo que es el factoring, el origen del mismo, su evolución y hace una clasificación de los distintos tipos de factoring.

  3. Characterization of the atmospheric pollution level in Sfax (Tunisia): influence of sources and meteorological factors; Caracterisation du niveau de pollution atmospherique dans la ville de Sfax (Tunisie): influence des sources et des facteurs meteorologiques

    Energy Technology Data Exchange (ETDEWEB)

    Azri, Ch.; Maalej, A.; Medhioub, K. [Ecole Nationale d' ingenieurs de Sfax, Unite de Recherche Etude et Gestion des Environnements Cotiers et Urbains (Tunisia); Tlili, A. [Faculte des Sciences de Sfax, Dept. de Geologie (Tunisia)

    2002-01-01

    This study held in Sfax City (Tunisia), showed that the atmospheric pollution level is strongly influenced by industrial sources, obstacles and meteorological factors. The factory of phosphate treatment 'SIAPE' displayed the main polluting source in the City. It is the principal issuing of SO{sub x} and toxic metals. The phosphogypsum deposit displayed an obstacle to the atmospheric diffusion of pollutants. Their stagnation is accentuated under smoky conditions. Several episodes of SO{sub 2} and dust are attributed to the effect of marked thermal inversions and the sirocco wind. The limited influence of traffic was proved by the registration of NO{sub x}, which showed low concentrations. It is conditioned by the change of fashion life of people and their customs of work (Ramadan month, agricultural activities). As a result, the adequate treatment of atmospheric industrial emanations (notably those of SIAPE) and the evacuation of artificial obstacles have to be considered. (authors)

  4. Characterization of calculation of in-situ retardation factors of contaminant transport using naturally-radionuclides and rock/water interaction occurring U-Series disequilibria timescales. 1997 annual progress report

    International Nuclear Information System (INIS)

    Goldstein, S.; Ku, T.L.; Luo, S.; Murrel, M.; Roback, R.

    1997-01-01

    'The research is directed toward a quantitative assessment of contaminant transport rates in fracture-rock systems using uranium-series radionuclides. Naturally occurring uranium-and thorium-series radioactive disequilibria will provide information on the rates of adsorption-desorption and transport of radioactive contaminants as well as on fluid transport and rock dissolution in a natural setting. This study will also provide an improved characterization of preferential flow and contaminant transport at the Idaho Environmental and Engineering Lab. (INEEL) site. To a lesser extent, the study will include rocks in the unsaturated zone. The authors will produce a realistic model of radionuclide migration under unsaturated and saturated field conditions at the INEEL site, taking into account the retardation processes involved in the rock/water interaction. The major tasks are to (1) determine the natural distribution of U, Th, Pa and Ra isotopes in rock minerals. sorbed phases on the rocks, and in fluids from both saturated and unsaturated zones at the site, and (2) study rock/water interaction processes using U/Th series disequilibrium and a statistical analysis-based model for the Geologic heterogeneity plays an important role in transporting contaminants in fractured rocks. Preferential flow paths in the fractured rocks act as a major pathway for transport of radioactive contaminants in groundwaters. The weathering/dissolution of rock by groundwater also influences contaminant mobility. Thus, it is important to understand the hydrogeologic features of the site and their impact on the migration of radioactive contaminants. In this regard, quantification of the rock weathering/dissolution rate and fluid residence time from the observed decay-series disequilibria will be valuable. By mapping the spatial distribution of the residence time of groundwater in fractured rocks, the subsurface preferential flow paths (with high rock permeability and short fluid residence

  5. Characterization and Ectopic Expression of CoWRI1, an AP2/EREBP Domain-Containing Transcription Factor from Coconut (Cocos nucifera L.) Endosperm, Changes the Seeds Oil Content in Transgenic Arabidopsis thaliana and Rice (Oryza sativa L.).

    Science.gov (United States)

    Sun, RuHao; Ye, Rongjian; Gao, Lingchao; Zhang, Lin; Wang, Rui; Mao, Ting; Zheng, Yusheng; Li, Dongdong; Lin, Yongjun

    2017-01-01

    Coconut ( Cocos nucifera L.) is a key tropical crop and a member of the monocotyledonous family Arecaceae ( Palmaceae ). Few genes and related metabolic processes involved in coconut endosperm development have been investigated. In this study, a new member of the WRI1 gene family was isolated from coconut endosperm and was named CoWRI1 . Its transcriptional activities and interactions with the acetyl-CoA carboxylase ( BCCP2 ) promoter of CoWRI1 were confirmed by the yeast two-hybrid and yeast one-hybrid approaches, respectively. Functional characterization was carried out through seed-specific expression in Arabidopsis and endosperm-specific expression in rice. In transgenic Arabidopsis , high over-expressions of CoWRI1 in seven independent T2 lines were detected by quantitative real-time PCR. The relative mRNA accumulation of genes encoding enzymes involved in either fatty acid biosynthesis or triacylglycerols assembly (BCCP2, KASI, MAT, ENR, FATA, and GPDH) were also assayed in mature seeds. Furthermore, lipid and fatty acids C16:0 and C18:0 significantly increased. In two homozygous T2 transgenic rice lines (G5 and G2), different CoWRI1 expression levels were detected, but no CoWRI1 transcripts were detected in the wild type. Analyses of the seed oil content, starch content, and total protein content indicated that the two T2 transgenic lines showed a significant increase ( P oil content. The transgenic lines also showed a significant increase in starch content, whereas total protein content decreased significantly. Further analysis of the fatty acid composition revealed that palmitic acid (C16:0) and linolenic acid (C18:3) increased significantly in the seeds of the transgenic rice lines, but oleic acid (C18:1) levels significantly declined.

  6. Characterization of novel heat-responsive transcription factor (TaHSFA6e) gene involved in regulation of heat shock proteins (HSPs) - A key member of heat stress-tolerance network of wheat.

    Science.gov (United States)

    Kumar, Ranjeet R; Goswami, Suneha; Singh, Khushboo; Dubey, Kavita; Rai, Gyanendra K; Singh, Bhupinder; Singh, Shivdhar; Grover, Monendra; Mishra, Dwijesh; Kumar, Sanjeev; Bakshi, Suman; Rai, Anil; Pathak, Himanshu; Chinnusamy, Viswanathan; Praveen, Shelly

    2018-08-10

    Heat stress has an adverse effect on the quality and quantity of agriculturally important crops, especially wheat. The tolerance mechanism has not been explored much in wheat and very few genes/ TFs responsive to heat stress is available on public domain. Here, we identified, cloned and characterized a putative TaHSFA6e TF gene of 1.3 kb from wheat cv. HD2985. We observed an ORF of 368 aa with Hsf DNA binding signature domain in the amino acid sequence. Single copy number of TaHSFA6e was observed integrated in the genome of wheat. Expression analysis of TaHSFA6e under differential HS showed maximum transcripts in wheat cv. Halna (thermotolerant) in response to 38 °C for 2 h during pollination and grain-filling stages, as compared to PBW343, HD2329 and HD2985. Putative target genes of TaHSFA6e (HSP17, HSP70 and HSP90) showed upregulation in response to differential HS (30 & 38 °C, 2 h) during pollination and grain-filling stages. Small HSP17 was observed most triggered in Halna under HS. We observed increase in the catalase, guaiacol peroxidase, total antioxidant capacity (TAC), and decrease in the lipid peroxidation in thermotolerant cvs. (Halna, HD2985), as compared to thermosusceptible (PBW343, HD2329) under differential HS. Multiple stresses (heat - 38 °C, 2 h, and drought - 100 mL of 20% polyethylene Glycol 6000) during seedling stage of wheat showed positive correlation between the expression of TaHSFA6e, putative targets (HSP70, HSP90, HSP17) and TAC. Halna (thermotolerant) performed better, as compared to other contrasting cvs. TaHSFA6e TF can be used as promising candidate gene for manipulating the heat stress-tolerance network. Copyright © 2018 Elsevier B.V. All rights reserved.

  7. Multi-faceted proteomic characterization of host protein complement of Rift Valley fever virus virions and identification of specific heat shock proteins, including HSP90, as important viral host factors.

    Science.gov (United States)

    Nuss, Jonathan E; Kehn-Hall, Kylene; Benedict, Ashwini; Costantino, Julie; Ward, Michael; Peyser, Brian D; Retterer, Cary J; Tressler, Lyal E; Wanner, Laura M; McGovern, Hugh F; Zaidi, Anum; Anthony, Scott M; Kota, Krishna P; Bavari, Sina; Hakami, Ramin M

    2014-01-01

    Rift Valley fever is a potentially fatal disease of humans and domestic animals caused by Rift Valley fever virus (RVFV). Infection with RVFV in ruminants can cause near 100% abortion rates and recent outbreaks in naïve human populations have suggested case fatality rates of greater than thirty percent. To elucidate the roles that host proteins play during RVFV infection, proteomic analysis of RVFV virions was conducted using complementary analytical approaches, followed by functional validation studies of select identified host factors. Coupling the more traditional Gel LC/MS/MS approach (SDS PAGE followed by liquid chromatography tandem mass spectrometry) with an alternative technique that preserves protein complexes allowed the protein complement of these viral particles to be thoroughly examined. In addition to viral proteins present within the virions and virion-associated host proteins, multiple macromolecular complexes were identified. Bioinformatic analysis showed that host chaperones were among over-represented protein families associated with virions, and functional experiments using siRNA gene silencing and small molecule inhibitors identified several of these heat shock proteins, including heat shock protein 90 (HSP90), as important viral host factors. Further analysis indicated that HSP inhibition effects occur during the replication/transcription phase of the virus life cycle, leading to significant lowering of viral titers without compromising the functional capacity of released virions. Overall, these studies provide much needed further insight into interactions between RVFV and host cells, increasing our understanding of the infection process and suggesting novel strategies for anti-viral development. In particular, considering that several HSP90 inhibitors have been advancing through clinical trials for cancer treatment, these results also highlight the exciting potential of repurposing HSP90 inhibitors to treat RVF.

  8. Multi-faceted proteomic characterization of host protein complement of Rift Valley fever virus virions and identification of specific heat shock proteins, including HSP90, as important viral host factors.

    Directory of Open Access Journals (Sweden)

    Jonathan E Nuss

    Full Text Available Rift Valley fever is a potentially fatal disease of humans and domestic animals caused by Rift Valley fever virus (RVFV. Infection with RVFV in ruminants can cause near 100% abortion rates and recent outbreaks in naïve human populations have suggested case fatality rates of greater than thirty percent. To elucidate the roles that host proteins play during RVFV infection, proteomic analysis of RVFV virions was conducted using complementary analytical approaches, followed by functional validation studies of select identified host factors. Coupling the more traditional Gel LC/MS/MS approach (SDS PAGE followed by liquid chromatography tandem mass spectrometry with an alternative technique that preserves protein complexes allowed the protein complement of these viral particles to be thoroughly examined. In addition to viral proteins present within the virions and virion-associated host proteins, multiple macromolecular complexes were identified. Bioinformatic analysis showed that host chaperones were among over-represented protein families associated with virions, and functional experiments using siRNA gene silencing and small molecule inhibitors identified several of these heat shock proteins, including heat shock protein 90 (HSP90, as important viral host factors. Further analysis indicated that HSP inhibition effects occur during the replication/transcription phase of the virus life cycle, leading to significant lowering of viral titers without compromising the functional capacity of released virions. Overall, these studies provide much needed further insight into interactions between RVFV and host cells, increasing our understanding of the infection process and suggesting novel strategies for anti-viral development. In particular, considering that several HSP90 inhibitors have been advancing through clinical trials for cancer treatment, these results also highlight the exciting potential of repurposing HSP90 inhibitors to treat RVF.

  9. Role of the 2 adenine (g.11293_11294insAA) insertion polymorphism in the 3' untranslated region of the factor VII (FVII) gene: molecular characterization of a patient with severe FVII deficiency.

    Science.gov (United States)

    Peyvandi, F; Garagiola, I; Palla, R; Marziliano, N; Mannucci, P M

    2005-11-01

    Polymorphic variants in the gene encoding factor VII (F7) affect the plasma levels of this coagulation protein and modify the clinical phenotype of FVII deficiency in some patients. In this study we report the in vitro functional analysis of a novel polymorphic variant located in the 3' untranslated region of F7: g.11293_11294insAA. To determine whether this variant regulates FVII expression, we initially compared an expression vector containing FVII cDNA with g.11293_11294insAA with the FVII wild-type (WT) construct. The kinetics of mRNA production showed that the insertion decreases the steady-state FVII mRNA levels. To assess whether the insertion influences the phenotype of FVII-deficient patients, we evaluated its effect on the expression of FVII in a patient with severe FVII deficiency (undetectable FVII activity and antigen) carrying two additional homozygous missense variations (p.Arg277Cys and p.Arg353Gln). The two substitutions alone reduced the expression of FVII activity and antigen in vitro, but with the insertion polymorphism in our expression vector the patient's phenotype of undetectable plasma FVII was recapitulated. The insertion polymorphism in the 3' untranslated region of F7 is another modifier of FVII expression that might explain the poor genotype-phenotype correlation in some FVII-deficient patients. Copyright 2005 Wiley-Liss, Inc.

  10. Nicotine can skew the characterization of the macrophage type-1 (M{Phi}1) phenotype differentiated with granulocyte-macrophage colony-stimulating factor to the M{Phi}2 phenotype

    Energy Technology Data Exchange (ETDEWEB)

    Yanagita, Manabu; Kobayashi, Ryohei [Department of Periodontology, Division of Oral Biology and Disease Control, Osaka University Graduate School of Dentistry, Osaka 565-0871 (Japan); Murakami, Shinya, E-mail: ipshinya@dent.osaka-u.ac.jp [Department of Periodontology, Division of Oral Biology and Disease Control, Osaka University Graduate School of Dentistry, Osaka 565-0871 (Japan)

    2009-10-09

    Macrophages (M{Phi}s) exhibit functional heterogeneity and plasticity in the local microenvironment. Recently, it was reported that M{Phi}s can be divided into proinflammatory M{Phi}s (M{Phi}1) and anti-inflammatory M{Phi}s (M{Phi}2) based on their polarized functional properties. Here, we report that nicotine, the major ingredient of cigarette smoke, can modulate the characteristics of M{Phi}1. Granulocyte-macrophage colony-stimulating factor-driven M{Phi}1 with nicotine (Ni-M{Phi}1) showed the phenotypic characteristics of M{Phi}2. Like M{Phi}2, Ni-M{Phi}1 exhibited antigen-uptake activities. Ni-M{Phi}1 suppressed IL-12, but maintained IL-10 and produced high amounts of MCP-1 upon lipopolysaccharide stimulation compared with M{Phi}1. Moreover, we observed strong proliferative responses of T cells to lipopolysaccharide-stimulated M{Phi}1, whereas Ni-M{Phi}1 reduced T cell proliferation and inhibited IFN-{gamma} production by T cells. These results suggest that nicotine can change the functional characteristics of M{Phi} and skew the M{Phi}1 phenotype to M{Phi}2. We propose that nicotine is a potent regulator that modulates immune responses in microenvironments.

  11. El factoring

    Directory of Open Access Journals (Sweden)

    Alberto Rosenthal

    2015-04-01

    Full Text Available RESUMEN Se presenta la segunda parte del artículo aparecido en  el número 6 de la revista EAN. Su contenido es complementario a lo expuesto en dicho número, en está aparecen las ventajas del factoring, conveniencias, limitaciones así como la forma  de efectuar un factor en Colombia,  su necesidad, incidencia económica, etc.

  12. Practical materials characterization

    CERN Document Server

    2014-01-01

    Presents cross-comparison between materials characterization techniquesIncludes clear specifications of strengths and limitations of each technique for specific materials characterization problemFocuses on applications and clear data interpretation without extensive mathematics

  13. In Vitro and In Vivo Characterization of a Dual-Function Green Fluorescent Protein–HSV1-Thymidine Kinase Reporter Gene Driven by the Human Elongation Factor 1α Promoter

    Directory of Open Access Journals (Sweden)

    Gary D. Luker

    2002-04-01

    Full Text Available Toward the goal of monitoring activity of native mammalian promoters with molecular imaging techniques, we stably transfected DU145 prostate carcinoma cells with a fusion construct of enhanced green fluorescent protein (EGFP and wild-type herpes simplex virus-1 thymidine kinase (HSV1-TK as a reporter gene driven by the promoter for human elongation factor 1α (EF-1α-EGFP-TK. Using this model system, expression of EGFP was quantified by flow cytometry and fluorescence microscopy, while the HSV1-TK component of the reporter was quantified with 8-[3H]ganciclovir (8-[3H]GCV. As analyzed by flow cytometry, passage of EGFP-TK-DU145 transfected cells (ETK in vitro resulted in populations of cells with high and low expression of EGFP over time. High and low ETK cells retained 23-fold and 5-fold more GCV, respectively, than control. While differences in uptake and retention of GCV corresponded to relative expression of the reporter gene in each subpopulation of cells as determined by both flow cytometry (EGFP and quantitative RT-PCR, the correlation was not linear. Furthermore, in high ETK cells, net retention of various radiolabeled nucleoside analogues varied; the rank order was 8-[3H]GCV < 9-(4-fluoro-3-hydroxymethylbutylguanine ([18F]FHBG ≈ 8-[3H]penciclovir (8-[3H]PCV < 2′-fluoro-2′-deoxy-5-iodouracil-beta-d-arabinofuranoside (2-[14C]FIAU. Xenograft tumors of ETK cells in vivo accumulated 2.5-fold more 8-[3H]GCV per gram of tissue and showed greater fluorescence from EGFP than control DU145 cells, demonstrating that the reporter gene functioned in vivo. These data extend previous reports by showing that a human promoter can be detected in vitro and in vivo with a dual-function reporter exploiting optical and radiotracer techniques.

  14. Production and characterization of recombinantly derived peptides and antibodies for accurate determinations of somatolactin, growth hormone and insulin-like growth factor-I in European sea bass (Dicentrarchus labrax).

    Science.gov (United States)

    de Celis, S Vega-Rubín; Gómez-Requeni, P; Pérez-Sánchez, J

    2004-12-01

    A specific radioimmunoassay (RIA) for European sea bass (Dicentrarchus labrax) growth hormone (GH) was developed and validated. For this purpose, a stable source of GH was produced by means of recombinant DNA technology in a bacteria system. The identity of the purified protein (ion exchange chromatography) was demonstrated by Western blot and a specific GH antiserum was raised in rabbit. In Western blot and RIA system, this antiserum recognized specifically native and recombinant GH, and it did not cross-react with fish prolactin (PRL) and somatolactin (SL). In a similar way, a specific polyclonal antiserum against the now available recombinant European sea bass SL was raised and used in the RIA system to a sensitivity of 0.3 ng/ml (90% of binding of tracer). Further, European sea bass insulin-like growth factor-I (IGF-I) was cloned and sequenced, and its high degree of identity with IGF-I peptides of barramundi, tuna, and sparid fish allowed the use of a commercial IGF-I RIA based on barramundi IGF-I antiserum. These assay tools assisted for the first time accurate determinations of SL and GH-IGF-I axis activity in a fish species of the Moronidae family. Data values were compared to those found with gilthead sea bream (Sparus aurata), which is currently used as a Mediterranean fish model for growth endocrinology studies. As a characteristic feature, the average concentration year round of circulating GH in growing mature males of European sea bass was higher than in gilthead sea bream. By contrast, the average concentration of circulating SL was lower. Concerning to circulating concentration of IGF-I, the measured plasma values for a given growth rate were also lower in European sea bass. These findings are discussed on the basis of a different energy status that might allowed a reduced but more continuous growth in European sea bass.

  15. What characterizes persons with poor mental health?

    DEFF Research Database (Denmark)

    Christensen, Anne Illemann; Davidsen, Michael; Kjøller, Mette

    2014-01-01

    analysed by means of logistic regression models. Results: Men and women with poor mental health are characterized by being single, having a long-term illness, not being able to rely on help from others in case of illness and by feeling that family and friends demand too much of them. Men with poor mental...... health were further characterized by being a heavy smoker, and having a BMI below 25. Women with poor mental health were further characterized by being 16-44 years old and sedentary in leisure time. CONCLUSIONS THE PREVALENCE OF POOR MENTAL HEALTH IS HIGHER AMONG WOMEN THAN MEN, AND DIFFERENT FACTORS...... CHARACTERIZE MEN AND WOMEN WITH POOR MENTAL HEALTH THE PRESENT FINDINGS SUPPORT THE NOTION THAT BOTH SOCIO-DEMOGRAPHICS AND LIFESTYLE FACTORS ARE INDEPENDENTLY RELATED WITH POOR MENTAL HEALTH WE SUGGEST TAKING INTO ACCOUNT ALL THESE AREAS OF LIFE WHEN PLANNING ACTIVITIES TO PREVENT POOR MENTAL HEALTH AND WHEN...

  16. Waste Characterization Methods

    Energy Technology Data Exchange (ETDEWEB)

    Vigil-Holterman, Luciana R. [Los Alamos National Lab. (LANL), Los Alamos, NM (United States); Naranjo, Felicia Danielle [Los Alamos National Lab. (LANL), Los Alamos, NM (United States)

    2016-02-02

    This report discusses ways to classify waste as outlined by LANL. Waste Generators must make a waste determination and characterize regulated waste by appropriate analytical testing or use of acceptable knowledge (AK). Use of AK for characterization requires several source documents. Waste characterization documentation must be accurate, sufficient, and current (i.e., updated); relevant and traceable to the waste stream’s generation, characterization, and management; and not merely a list of information sources.

  17. Waste Characterization Methods

    International Nuclear Information System (INIS)

    Vigil-Holterman, Luciana R.; Naranjo, Felicia Danielle

    2016-01-01

    This report discusses ways to classify waste as outlined by LANL. Waste Generators must make a waste determination and characterize regulated waste by appropriate analytical testing or use of acceptable knowledge (AK). Use of AK for characterization requires several source documents. Waste characterization documentation must be accurate, sufficient, and current (i.e., updated); relevant and traceable to the waste stream's generation, characterization, and management; and not merely a list of information sources.

  18. 巴西蕉2个ERF转录因子基因的克隆及功能研究%Cloning and characterization of two ERF transcription factor genes in Brazil banana

    Institute of Scientific and Technical Information of China (English)

    侯晓婉; 胡伟; 徐碧玉; 张鲁斌; 金志强

    2017-01-01

    [Objective] Banana (Musaceae,Musa) is one of the most important food plants,widely distributing in tropical and subtropical countries.Banana is particularly sensitive to most of the abiotic stresses,such as freezing,drought and salt,which would cause heave decrease of the yield and quality.Therefore,investigation the molecular mechanism of the response of banana to abiotic stresses is of prime importance for improving stress-resistant ability.Moreover,over the last decades APETALA2/Ethylene Responsive Factor (AP2/ERF) proteins have been found to be involved in a variety of biological processes.Based on the number of the structure domain,AP2/ERF superfamily was divided into four families as AP2/ERF,ERF,RAV and soloist.The proteins of ERF family only contained an AP2/ERF structure domain and were studied more deeply than others in all transcription factors family.The genes of ERF family played an important role in regulating plant response to abiotic stresses.The purpose of this study is to clone the key ERF transcription factors of banana and to identify their functions in response to abiotic stresses in order to provide a basis for improving stress-resistant ability.[Methods] The relative transcriptome results of AP2/ERF superfamily genes of Brazilian banana involved in responding to abiotic stresses of leaves were obtained.The expression profile analysis on AP2/ERF super family genes of banana were conducted by using MeV software.In view of the sensitivity to low temperature stress,MaERF25 and MaERF27 were isolated from banana using RNA reverse transcription cDNA mixture from various tissues and organs of banana as template.By using ExPASy,DNAMAN and some other related biological analysis softwares and on the basis of protein sequence homology,the sequences homologue and the basic characteristics of protein were analyzed.In addition,the transient expression vectors containing MaERF25 and MaERF 27gene were constructed on the basis of plant expression vector p

  19. Quality factors

    International Nuclear Information System (INIS)

    Kerr, G.D.

    1986-01-01

    The quality factor, Q, is a dimensionless modifier used in converting absorbed dose, expressed in rads (or grays), to dose equivalent, expressed in rems (or seiverts). The dose equivalent is used in radiation protection to account for the biological effectiveness of different kinds of radiation. The quality factor is related to both the linear energy transfer (LET) and relative biological effectiveness (RBE). The RBE's obtained from biological experiments depend in a complex way on the observed biological effect, the specific test organism, and the experimental conditions. Judgement is involved, therefore, in the choice of the quality factor. Questions regarding the adequacy of current Q values for neutrons were raised first in a 1980 statement by the National Council on Radiation Protection (NCRP) and later in a 1985 statement by the International Commission on Radiological Protection (ICRP). In 1980, the NCRP alerted the technical community to possible future increases between a factor of three and ten in the Q for neutrons, and in 1985, the ICRP suggested an increase by a factor of two in Q for neutrons. Both the ICRP and NRCP are now recommending essentially the same guidance with regard to Q for neutrons: an increase by a factor of two. The Q for neutrons is based on a large, albeit unfocused, body of experimental data. In spite of the lack of focus, the data supporting a change in the neutron quality factor are substantial. However, the proposed doubling of Q for neutrons is clouded by other issues regarding its application. 33 refs., 4 figs., 6 tabs

  20. Discovery and Characterization of a Novel Lachrymatory Factor Synthase in Petiveria alliacea and Its Influence on Alliinase-Mediated Formation of Biologically Active Organosulfur Compounds1[W][OA

    Science.gov (United States)

    Musah, Rabi A.; He, Quan; Kubec, Roman

    2009-01-01

    A novel lachrymatory factor synthase (LFS) was isolated and purified from the roots of the Amazonian medicinal plant Petiveria alliacea. The enzyme is a heterotetrameric glycoprotein comprised of two α-subunits (68.8 kD each), one γ-subunit (22.5 kD), and one δ-subunit (11.9 kD). The two α-subunits are glycosylated and connected by a disulfide bridge. The LFS has an isoelectric point of 5.2. It catalyzes the formation of a sulfine lachrymator, (Z)-phenylmethanethial S-oxide, only in the presence of P. alliacea alliinase and its natural substrate, S-benzyl-l-cysteine sulfoxide (petiveriin). Depending on its concentration relative to that of P. alliacea alliinase, the LFS sequesters, to varying degrees, the sulfenic acid intermediate formed by alliinase-mediated breakdown of petiveriin. At LFS:alliinase of 5:1, LFS sequesters all of the sulfenic acid formed by alliinase action on petiveriin, and converts it entirely to (Z)-phenylmethanethial S-oxide. However, starting at LFS:alliinase of 5:2, the LFS is unable to sequester all of the sulfenic acid produced by the alliinase, with the result that sulfenic acid that escapes the action of the LFS condenses with loss of water to form S-benzyl phenylmethanethiosulfinate (petivericin). The results show that the LFS and alliinase function in tandem, with the alliinase furnishing the sulfenic acid substrate on which the LFS acts. The results also show that the LFS modulates the formation of biologically active thiosulfinates that are downstream of the alliinase in a manner dependent upon the relative concentrations of the LFS and the alliinase. These observations suggest that manipulation of LFS-to-alliinase ratios in plants displaying this system may provide a means by which to rationally modify organosulfur small molecule profiles to obtain desired flavor and/or odor signatures, or increase the presence of desirable biologically active small molecules. PMID:19692535

  1. Molecular characterization of FXI deficiency.

    Science.gov (United States)

    Berber, Ergul

    2011-02-01

    Factor XI (FXI) deficiency is a rare autosomal bleeding disease associated with genetic defects in the FXI gene. It is a heterogeneous disorder with variable tendency in bleeding and variable causative FXI gene mutations. It is characterized as a cross-reacting material-negative (CRM-) FXI deficiency due to decreased FXI levels or cross-reacting material-positive (CRM+) FXI deficiency due to impaired FXI function. Increasing number of mutations has been reported in FXI mutation database, and most of the mutations are affecting serine protease (SP) domain of the protein. Functional characterization for the mutations helps to better understand the molecular basis of FXI deficiency. Prevalence of the disease is higher in certain populations such as Ashkenazi Jews. The purpose of this review is to give an overview of the molecular basis of congenital FXI deficiency.

  2. High quality-factor optical resonators

    International Nuclear Information System (INIS)

    Henriet, Rémi; Salzenstein, Patrice; Coillet, Aurélien; Saleh, Khaldoun; Chembo, Yanne K; Ristic, Davor; Ferrari, Maurizio; Mortier, Michel; Rasoloniaina, Alphonse; Dumeige, Yannick; Féron, Patrice; Cibiel, Gilles; Llopis, Olivier

    2014-01-01

    Various resonators are investigated for microwave photonic applications. Micro-sphere, disk and fiber ring resonators were designed, realized and characterized. Obtained quality factors are as high as Q = 10 10 . (paper)

  3. Epidermal growth factor in the rat prostate

    DEFF Research Database (Denmark)

    Tørring, Niels; Jørgensen, P E; Poulsen, Steen Seier

    1998-01-01

    Epidermal growth factor (EGF) induces proliferation in prostate epithelial and stromal cells in primary culture. This investigation was set up to characterize the time and spatial expression of EGF in the rat prostate.......Epidermal growth factor (EGF) induces proliferation in prostate epithelial and stromal cells in primary culture. This investigation was set up to characterize the time and spatial expression of EGF in the rat prostate....

  4. Caracterización de los factores de riesgo en gestantes con hipertensión gestacional y crónica en un área de salud Characterization of the risk factors present in pregnant with chronic and gestational hypertension in a health area

    Directory of Open Access Journals (Sweden)

    Yuselis Torres Sánchez

    2009-09-01

    Full Text Available INTRODUCCIÓN: los trastornos hipertensivos de la gestación constituyen una de las complicaciones más frecuentes que causan morbimortalidad materno-fetal, por lo que la reducción de su prevalencia e incidencia es una prioridad. Para enfrentar este reto se debe tener un enfoque médico integral en la atención preconcepcional y concepcional a la mujer, que condicione una mejor calidad de vida. OBJETIVO: describir la tendencia que han tenido estos trastornos en los últimos 4 años (2004-2007, y los factores de riesgo que con más frecuencia se presentaron. MÉTODOS: se diseñó un estudio observacional analítico transversal utilizando un universo de estudio de 25 gestantes con trastornos hipertensivos de la gestación del área de salud "Dr. Antonio Pulido Humarán". RESULTADOS: la hipertensión gestacional y la hipertensión arterial crónica tuvieron una tendencia ascendente en su prevalencia en el período 2004-2007. La obesidad, el tabaquismo, las inadecuadas condiciones socioeconómicas, la no suplementación de folatos, entre otras, constituyeron los factores de riesgo más frecuentes.INTRODUCTION: pregnancy hypertensive disorders are one of the more frequent complications causing morbidity and mortality in mother-fetus, where decrease of its prevalence and incidence is a priority. To confront this challenge we must to have a integral medical approach on woman pre-conception and conception care to achieve a better quality of life.] OBJECTIVE: to describe trend of these disorders during past 4 years (2004-2007, and the more frequent risk factors. METHODS: authors designed a cross-sectional, analytical and descriptive study using a cohort of 25 pregnant with hypertensive disorders related to this condition from "Dr. Antonio Pulido Humarán" health area. RESULTS: pregnancy hypertension and chronic high blood pressure had a rising trend in its prevalence during 2004-2007. Obesity, smoking, and the inappropriate socioeconomic conditions, the

  5. Risk factors

    International Nuclear Information System (INIS)

    Dennery, M.; Dupont, M.A.

    2007-01-01

    This article deals with the development of risk management in the gas sector business: why a risk factor legal mention must precede any published financial information? Do gas companies have to face new risks? Is there specific risks bound to gas activities? Why companies want to master their risks? Is it mandatory or just a new habit? Do they expect a real benefit in return? These are the risk management questions that are analyzed in this article which is based on the public communication of 15 gas companies randomly selected over the world. The information comes from their annual reports or from documents available on their web sites. The intention of this document is not to be exhaustive or to make statistics but only to shade light on the risk factors of the gas sector. (J.S.)

  6. Organizational factors

    International Nuclear Information System (INIS)

    Wahlstroem, B.; Kettunen, J.

    2005-01-01

    The goal of this lecture is to give an overview of important concepts connected to organisational factors and to provide an understanding of mechanisms by which they can contribute to safe or unsafe behaviour of people. The lecture gives examples of ways to organise work, organisational deficiencies and good practices applied in safety oriented organisations. The lecture also gives an introduction to international work and Finnish national regulation connected to organisation and management. (orig.)

  7. Factor analysis

    CERN Document Server

    Gorsuch, Richard L

    2013-01-01

    Comprehensive and comprehensible, this classic covers the basic and advanced topics essential for using factor analysis as a scientific tool in psychology, education, sociology, and related areas. Emphasizing the usefulness of the techniques, it presents sufficient mathematical background for understanding and sufficient discussion of applications for effective use. This includes not only theory but also the empirical evaluations of the importance of mathematical distinctions for applied scientific analysis.

  8. Photovoltaic Characterization Laboratory

    Data.gov (United States)

    Federal Laboratory Consortium — NIST's PV characterization laboratory is used to measure the electrical performance and opto-electronic properties of solar cells and