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Sample records for addition high expression

  1. High expression Zymomonas promoters

    DOEpatents

    Viitanen, Paul V.; Tao, Luan; Zhang, Yuying; Caimi, Perry G.; McCole, Laura : Zhang, Min; Chou, Yat-Chen; McCutchen, Carol M.; Franden, Mary Ann

    2011-08-02

    Identified are mutants of the promoter of the Z. mobilis glyceraldehyde-3-phosphate dehydrogenase gene, which direct improved expression levels of operably linked heterologous nucleic acids. These are high expression promoters useful for expression of chimeric genes in Zymomonas, Zymobacter, and other related bacteria.

  2. Tough, High-Performance, Thermoplastic Addition Polymers

    NASA Technical Reports Server (NTRS)

    Pater, Ruth H.; Proctor, K. Mason; Gleason, John; Morgan, Cassandra; Partos, Richard

    1991-01-01

    Series of addition-type thermoplastics (ATT's) exhibit useful properties. Because of their addition curing and linear structure, ATT polymers have toughness, like thermoplastics, and easily processed, like thermosets. Work undertaken to develop chemical reaction forming stable aromatic rings in backbone of ATT polymer, combining high-temperature performance and thermo-oxidative stability with toughness and easy processibility, and minimizing or eliminating necessity for tradeoffs among properties often observed in conventional polymer syntheses.

  3. Additives for high-temperature liquid lubricants

    NASA Technical Reports Server (NTRS)

    Lawton, Emil A.; Yavrouian, Andre H.; Repar, John

    1988-01-01

    A preliminary research program was conducted to demonstrate a new concept for additives to liquid lubricants. It was demonstrated that suspensions of o-phthalonitrile and a substituted 1,2-maleonitrile in mineral oil and dilute solutions of o-phthalonitrile and tetrafluoro-o-phthalonitrile extended the lifetime of bearings under boundary lubricating conditions. The solutions exhibited coefficients of friction under high loads of 0.02-0.03. These results were consistent with the hypothesis that these compounds react with the hot metal surface to form a planar lubricating film by means of a metal or metal oxide template reaction. Also, the adherence was very strong due to the chelating action of the planar macrocycles postulated to form under the experimental conditions.

  4. Additive Expression of Consolidated Memory through Drosophila Mushroom Body Subsets.

    PubMed

    Yang, Chu-Huai; Shih, Meng-Fu Maxwell; Chang, Ching-Ching; Chiang, Meng-Hsuan; Shih, Hsiang-Wen; Tsai, Ya-Lun; Chiang, Ann-Shyn; Fu, Tsai-Feng; Wu, Chia-Lin

    2016-05-01

    Associative olfactory memory in Drosophila has two components called labile anesthesia-sensitive memory and consolidated anesthesia-resistant memory (ARM). Mushroom body (MB) is a brain region critical for the olfactory memory and comprised of 2000 neurons that can be classified into αβ, α'β', and γ neurons. Previously we demonstrated that two parallel pathways mediated ARM consolidation: the serotonergic dorsal paired medial (DPM)-αβ neurons and the octopaminergic anterior paired lateral (APL)-α'β' neurons. This finding prompted us to ask how this composite ARM is retrieved. Here, we showed that blocking the output of αβ neurons and that of α'β' neurons each impaired ARM retrieval, and blocking both simultaneously had an additive effect. Knockdown of radish and octβ2R in αβ and α'β' neurons, respectively, impaired ARM. A combinatorial assay of radish mutant background rsh1 and neurotransmission blockade confirmed that ARM retrieved from α'β' neuron output is independent of radish. We identified MBON-β2β'2a and MBON-β'2mp as the MB output neurons downstream of αβ and α'β' neurons, respectively, whose glutamatergic transmissions also additively contribute to ARM retrieval. Finally, we showed that α'β' neurons could be functionally subdivided into α'β'm neurons required for ARM retrieval, and α'β'ap neurons required for ARM consolidation. Our work demonstrated that two parallel neural pathways mediating ARM consolidation in Drosophila MB additively contribute to ARM expression during retrieval.

  5. Additives

    NASA Technical Reports Server (NTRS)

    Smalheer, C. V.

    1973-01-01

    The chemistry of lubricant additives is discussed to show what the additives are chemically and what functions they perform in the lubrication of various kinds of equipment. Current theories regarding the mode of action of lubricant additives are presented. The additive groups discussed include the following: (1) detergents and dispersants, (2) corrosion inhibitors, (3) antioxidants, (4) viscosity index improvers, (5) pour point depressants, and (6) antifouling agents.

  6. EAST ELEVATION OF HIGH BAY ADDITION OF FUEL STORAGE BUILDING ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    EAST ELEVATION OF HIGH BAY ADDITION OF FUEL STORAGE BUILDING (CPP-603). INL DRAWING NUMBER 200-0603-00-706-051286. - Idaho National Engineering Laboratory, Idaho Chemical Processing Plant, Fuel Reprocessing Complex, Scoville, Butte County, ID

  7. Tough, high performance, addition-type thermoplastic polymers

    NASA Technical Reports Server (NTRS)

    Pater, Ruth H. (Inventor)

    1992-01-01

    A tough, high performance polyimide is provided by reacting a triple bond conjugated with an aromatic ring in a bisethynyl compound with the active double bond in a compound containing a double bond activated toward the formation of a Diels-Adler type adduct, especially a bismaleimide, a biscitraconimide, or a benzoquinone, or mixtures thereof. Addition curing of this product produces a high linear polymeric structure and heat treating the highly linear polymeric structure produces a thermally stable aromatic addition-type thermoplastic polyimide, which finds utility in the preparation of molding compounds, adhesive compositions, and polymer matrix composites.

  8. Additional disulfide bonds in insulin: Prediction, recombinant expression, receptor binding affinity, and stability

    PubMed Central

    Vinther, Tine N; Pettersson, Ingrid; Huus, Kasper; Schlein, Morten; Steensgaard, Dorte B; Sørensen, Anders; Jensen, Knud J; Kjeldsen, Thomas; Hubalek, František

    2015-01-01

    The structure of insulin, a glucose homeostasis-controlling hormone, is highly conserved in all vertebrates and stabilized by three disulfide bonds. Recently, we designed a novel insulin analogue containing a fourth disulfide bond located between positions A10-B4. The N-terminus of insulin's B-chain is flexible and can adapt multiple conformations. We examined how well disulfide bond predictions algorithms could identify disulfide bonds in this region of insulin. In order to identify stable insulin analogues with additional disulfide bonds, which could be expressed, the Cβ cut-off distance had to be increased in many instances and single X-ray structures as well as structures from MD simulations had to be used. The analogues that were identified by the algorithm without extensive adjustments of the prediction parameters were more thermally stable as assessed by DSC and CD and expressed in higher yields in comparison to analogues with additional disulfide bonds that were more difficult to predict. In contrast, addition of the fourth disulfide bond rendered all analogues resistant to fibrillation under stress conditions and all stable analogues bound to the insulin receptor with picomolar affinities. Thus activity and fibrillation propensity did not correlate with the results from the prediction algorithm. PMID:25627966

  9. Additives for high temperature liquid lubricants. Final report

    SciTech Connect

    Yavrouian, A.H.; Repar, J.; Moran, C.M.; Lawton, E.A.; Anderson, M.S.

    1994-01-15

    The purpose of this task was to perform research for the Department of Energy (DOE) on the synthesis and characterization of additives for liquid lubricants which could lead to significant improvements in the major tribological task area of friction and wear reduction at high temperature. To this end JPL surveyed candidate precursor compounds which are soluble in liquid lubricants, synthesized the most promising of these materials, characterized them and submitted these additives to National Institute of Standards and Technology (NIST) for evaluation.

  10. High School Science Technology Additions, Midland Public Schools.

    ERIC Educational Resources Information Center

    Design Cost Data, 2001

    2001-01-01

    Discusses design goals, space requirements, and need for mobile furniture and "imagination stations" at Michigan's Midland Public High School science technology addition. Describes the architectural design, costs, and specifications. Includes floor plans, general description, photos and a list of consultants, manufacturers, and suppliers used for…

  11. Additive Manufacturing of High-Entropy Alloys by Laser Processing

    NASA Astrophysics Data System (ADS)

    Ocelík, V.; Janssen, N.; Smith, S. N.; De Hosson, J. Th. M.

    2016-07-01

    This contribution concentrates on the possibilities of additive manufacturing of high-entropy clad layers by laser processing. In particular, the effects of the laser surface processing parameters on the microstructure and hardness of high-entropy alloys (HEAs) were examined. AlCoCrFeNi alloys with different amounts of aluminum prepared by arc melting were investigated and compared with the laser beam remelted HEAs with the same composition. Attempts to form HEAs coatings with a direct laser deposition from the mixture of elemental powders were made for AlCoCrFeNi and AlCrFeNiTa composition. A strong influence of solidification rate on the amounts of face-centered cubic and body-centered cubic phase, their chemical composition, and spatial distribution was detected for two-phase AlCoCrFeNi HEAs. It is concluded that a high-power laser is a versatile tool to synthesize interesting HEAs with additive manufacturing processing. Critical issues are related to the rate of (re)solidification, the dilution with the substrate, powder efficiency during cladding, and differences in melting points of clad powders making additive manufacturing processing from a simple mixture of elemental powders a challenging approach.

  12. Highly oriented carbon fiber–polymer composites via additive manufacturing

    SciTech Connect

    Tekinalp, Halil L.; Kunc, Vlastimil; Velez-Garcia, Gregorio M.; Duty, Chad E.; Love, Lonnie J.; Naskar, Amit K.; Blue, Craig A.; Ozcan, Soydan

    2014-10-16

    Additive manufacturing, diverging from traditional manufacturing techniques, such as casting and machining materials, can handle complex shapes with great design flexibility without the typical waste. Although this technique has been mainly used for rapid prototyping, interest is growing in using this method to directly manufacture actual parts of complex shape. To use 3D-printing additive manufacturing in wide spread applications, the technique and the feedstock materials require improvements to meet the mechanical requirements of load-bearing components. Thus, we investigated the short fiber (0.2 mm to 0.4 mm) reinforced acrylonitrile-butadiene-styrene composites as a feedstock for 3D-printing in terms of their processibility, microstructure and mechanical performance; and also provided comparison with traditional compression molded composites. The tensile strength and modulus of 3D-printed samples increased ~115% and ~700%, respectively. 3D-printer yielded samples with very high fiber orientation in printing direction (up to 91.5 %), whereas, compression molding process yielded samples with significantly less fiber orientation. Microstructure-mechanical property relationships revealed that although the relatively high porosity is observed in the 3D-printed composites as compared to those produced by the conventional compression molding technique, they both exhibited comparable tensile strength and modulus. Furthermore, this phenomena is explained based on the changes in fiber orientation, dispersion and void formation.

  13. Highly oriented carbon fiber–polymer composites via additive manufacturing

    DOE PAGES

    Tekinalp, Halil L.; Kunc, Vlastimil; Velez-Garcia, Gregorio M.; Duty, Chad E.; Love, Lonnie J.; Naskar, Amit K.; Blue, Craig A.; Ozcan, Soydan

    2014-10-16

    Additive manufacturing, diverging from traditional manufacturing techniques, such as casting and machining materials, can handle complex shapes with great design flexibility without the typical waste. Although this technique has been mainly used for rapid prototyping, interest is growing in using this method to directly manufacture actual parts of complex shape. To use 3D-printing additive manufacturing in wide spread applications, the technique and the feedstock materials require improvements to meet the mechanical requirements of load-bearing components. Thus, we investigated the short fiber (0.2 mm to 0.4 mm) reinforced acrylonitrile-butadiene-styrene composites as a feedstock for 3D-printing in terms of their processibility, microstructuremore » and mechanical performance; and also provided comparison with traditional compression molded composites. The tensile strength and modulus of 3D-printed samples increased ~115% and ~700%, respectively. 3D-printer yielded samples with very high fiber orientation in printing direction (up to 91.5 %), whereas, compression molding process yielded samples with significantly less fiber orientation. Microstructure-mechanical property relationships revealed that although the relatively high porosity is observed in the 3D-printed composites as compared to those produced by the conventional compression molding technique, they both exhibited comparable tensile strength and modulus. Furthermore, this phenomena is explained based on the changes in fiber orientation, dispersion and void formation.« less

  14. Additivity principle in high-dimensional deterministic systems.

    PubMed

    Saito, Keiji; Dhar, Abhishek

    2011-12-16

    The additivity principle (AP), conjectured by Bodineau and Derrida [Phys. Rev. Lett. 92, 180601 (2004)], is discussed for the case of heat conduction in three-dimensional disordered harmonic lattices to consider the effects of deterministic dynamics, higher dimensionality, and different transport regimes, i.e., ballistic, diffusive, and anomalous transport. The cumulant generating function (CGF) for heat transfer is accurately calculated and compared with the one given by the AP. In the diffusive regime, we find a clear agreement with the conjecture even if the system is high dimensional. Surprisingly, even in the anomalous regime the CGF is also well fitted by the AP. Lower-dimensional systems are also studied and the importance of three dimensionality for the validity is stressed. PMID:22243060

  15. Random sequential addition of hard spheres in high Euclidean dimensions

    NASA Astrophysics Data System (ADS)

    Torquato, S.; Uche, O. U.; Stillinger, F. H.

    2006-12-01

    Sphere packings in high dimensions have been the subject of recent theoretical interest. Employing numerical and theoretical methods, we investigate the structural characteristics of random sequential addition (RSA) of congruent spheres in d -dimensional Euclidean space Rd in the infinite-time or saturation limit for the first six space dimensions (1≤d≤6) . Specifically, we determine the saturation density, pair correlation function, cumulative coordination number and the structure factor in each of these dimensions. We find that for 2≤d≤6 , the saturation density ϕs scales with dimension as ϕs=c1/2d+c2d/2d , where c1=0.202048 and c2=0.973872 . We also show analytically that the same density scaling is expected to persist in the high-dimensional limit, albeit with different coefficients. A byproduct of this high-dimensional analysis is a relatively sharp lower bound on the saturation density for any d given by ϕs≥(d+2)(1-S0)/2d+1 , where S0ɛ[0,1] is the structure factor at k=0 (i.e., infinite-wavelength number variance) in the high-dimensional limit. We demonstrate that a Palàsti-type conjecture (the saturation density in Rd is equal to that of the one-dimensional problem raised to the d th power) cannot be true for RSA hyperspheres. We show that the structure factor S(k) must be analytic at k=0 and that RSA packings for 1≤d≤6 are nearly “hyperuniform.” Consistent with the recent “decorrelation principle,” we find that pair correlations markedly diminish as the space dimension increases up to six. We also obtain kissing (contact) number statistics for saturated RSA configurations on the surface of a d -dimensional sphere for dimensions 2≤d≤5 and compare to the maximal kissing numbers in these dimensions. We determine the structure factor exactly for the related “ghost” RSA packing in Rd and demonstrate that its distance from “hyperuniformity” increases as the space dimension increases, approaching a constant asymptotic value

  16. Expressive Movement of High School Choral Musicians

    ERIC Educational Resources Information Center

    Rohwer, Debbie; Rohwer, Mark

    2014-01-01

    There is a need for a musical ensemble study that can describe the idiosyncratic, authentic movements that choral musicians make in a performance setting. In addition, it could benefit teachers to know whether those who score highest on a measure of expressive performance also tend to be strong performers. If this is the case, then the link…

  17. Asymmetric Uncertainty Expression for High Gradient Aerodynamics

    NASA Technical Reports Server (NTRS)

    Pinier, Jeremy T

    2012-01-01

    When the physics of the flow around an aircraft changes very abruptly either in time or space (e.g., flow separation/reattachment, boundary layer transition, unsteadiness, shocks, etc), the measurements that are performed in a simulated environment like a wind tunnel test or a computational simulation will most likely incorrectly predict the exact location of where (or when) the change in physics happens. There are many reasons for this, includ- ing the error introduced by simulating a real system at a smaller scale and at non-ideal conditions, or the error due to turbulence models in a computational simulation. The un- certainty analysis principles that have been developed and are being implemented today do not fully account for uncertainty in the knowledge of the location of abrupt physics changes or sharp gradients, leading to a potentially underestimated uncertainty in those areas. To address this problem, a new asymmetric aerodynamic uncertainty expression containing an extra term to account for a phase-uncertainty, the magnitude of which is emphasized in the high-gradient aerodynamic regions is proposed in this paper. Additionally, based on previous work, a method for dispersing aerodynamic data within asymmetric uncer- tainty bounds in a more realistic way has been developed for use within Monte Carlo-type analyses.

  18. Clustering of High Throughput Gene Expression Data

    PubMed Central

    Pirim, Harun; Ekşioğlu, Burak; Perkins, Andy; Yüceer, Çetin

    2012-01-01

    High throughput biological data need to be processed, analyzed, and interpreted to address problems in life sciences. Bioinformatics, computational biology, and systems biology deal with biological problems using computational methods. Clustering is one of the methods used to gain insight into biological processes, particularly at the genomics level. Clearly, clustering can be used in many areas of biological data analysis. However, this paper presents a review of the current clustering algorithms designed especially for analyzing gene expression data. It is also intended to introduce one of the main problems in bioinformatics - clustering gene expression data - to the operations research community. PMID:23144527

  19. System and method for high power diode based additive manufacturing

    DOEpatents

    El-Dasher, Bassem S.; Bayramian, Andrew; Demuth, James A.; Farmer, Joseph C.; Torres, Sharon G.

    2016-04-12

    A system is disclosed for performing an Additive Manufacturing (AM) fabrication process on a powdered material forming a substrate. The system may make use of a diode array for generating an optical signal sufficient to melt a powdered material of the substrate. A mask may be used for preventing a first predetermined portion of the optical signal from reaching the substrate, while allowing a second predetermined portion to reach the substrate. At least one processor may be used for controlling an output of the diode array.

  20. Processable high temperature resistant addition type polyimide laminating resins

    NASA Technical Reports Server (NTRS)

    Serafini, T. T.; Delvigs, P.

    1973-01-01

    Basic studies that were performed using model compounds to elucidate the polymerization mechanism of the so-called addition-type (A-type) polyimides are reviewed. The fabrication and properties of polyimide/graphite fiber composites using A-type polyimide prepolymers as the matrix are also reviewed. An alternate method for preparing processable A-type polyimides by means of in situ polymerization of monomer reactants (PMR) on the fiber reinforcement is described. The elevated temperature properties of A-type PMR/graphite fiber composites are also presented.

  1. Solvent additive to achieve highly ordered nanostructural semicrystalline DPP copolymers: toward a high charge carrier mobility.

    PubMed

    An, Tae Kyu; Kang, Il; Yun, Hui-jun; Cha, Hyojung; Hwang, Jihun; Park, Seonuk; Kim, Jiye; Kim, Yu Jin; Chung, Dae Sung; Kwon, Soon-Ki; Kim, Yun-Hi; Park, Chan Eon

    2013-12-23

    A facile spin-coating method in which a small percentage of the solvent additive, 1-chloronaphthalene (CN), is found to increase the drying time during film deposition, is reported. The field-effect mobility of a PDPPDBTE film cast from a chloroform-CN mixed solution is 0.46 cm(2) V(-1) s(-1). The addition of CN to the chloroform solution facilitates the formation of highly crystalline polymer structures. PMID:24115273

  2. Emotional expression in music: contribution, linearity, and additivity of primary musical cues.

    PubMed

    Eerola, Tuomas; Friberg, Anders; Bresin, Roberto

    2013-01-01

    The aim of this study is to manipulate musical cues systematically to determine the aspects of music that contribute to emotional expression, and whether these cues operate in additive or interactive fashion, and whether the cue levels can be characterized as linear or non-linear. An optimized factorial design was used with six primary musical cues (mode, tempo, dynamics, articulation, timbre, and register) across four different music examples. Listeners rated 200 musical examples according to four perceived emotional characters (happy, sad, peaceful, and scary). The results exhibited robust effects for all cues and the ranked importance of these was established by multiple regression. The most important cue was mode followed by tempo, register, dynamics, articulation, and timbre, although the ranking varied across the emotions. The second main result suggested that most cue levels contributed to the emotions in a linear fashion, explaining 77-89% of variance in ratings. Quadratic encoding of cues did lead to minor but significant increases of the models (0-8%). Finally, the interactions between the cues were non-existent suggesting that the cues operate mostly in an additive fashion, corroborating recent findings on emotional expression in music (Juslin and Lindström, 2010). PMID:23908642

  3. High-Flow, High-Molecular-Weight, Addition-Curing Polyimides

    NASA Technical Reports Server (NTRS)

    Chuang, Kathy C.; Vannucci, Raymond D.

    1993-01-01

    In developed series of high-flow PMR-type polyimide resins, 2, 2'-bis(trifluoromethyl)-4, 4'-diaminobiphenyl (BTDB) substituted for 1, 4-pheylenediamine in PMR-II formulation. Polyimides designated either as PMR-12F when nadic ester (NE) end caps used, or as V-CAP-12F when p-aminostyrene end caps used. High-molecular-weight, addition-curing polyimides based on BTBD and HFDE highly processable high-temperature matrix resins used to make composite materials with excellent retention of properties during long-term exposure to air at 650 degrees F or higher temperature. Furthermore, 12F addition-curing polyimides useful for electronic applications; fluorinated rigid-rod polyimides known to exhibit low thermal expansion coefficients as well as low absorption of moisture.

  4. Developing a High Thermal Conductivity Fuel with Silicon Carbide Additives

    SciTech Connect

    baney, Ronald; Tulenko, James

    2012-11-20

    The objective of this research is to increase the thermal conductivity of uranium oxide (UO{sub 2}) without significantly impacting its neutronic properties. The concept is to incorporate another high thermal conductivity material, silicon carbide (SiC), in the form of whiskers or from nanoparticles of SiC and a SiC polymeric precursor into UO{sub 2}. This is expected to form a percolation pathway lattice for conductive heat transfer out of the fuel pellet. The thermal conductivity of SiC would control the overall fuel pellet thermal conductivity. The challenge is to show the effectiveness of a low temperature sintering process, because of a UO{sub 2}-SiC reaction at 1,377°C, a temperature far below the normal sintering temperature. Researchers will study three strategies to overcome the processing difficulties associated with pore clogging and the chemical reaction of SiC and UO{sub 2} at temperatures above 1,300°C:

  5. High-throughput insect cell protein expression applications.

    PubMed

    Buchs, Mirjam; Kim, Ernie; Pouliquen, Yann; Sachs, Michael; Geisse, Sabine; Mahnke, Marion; Hunt, Ian

    2009-01-01

    The Baculovirus Expression Vector System (BEVS) is one of the most efficient systems for production of recombinant proteins and consequently its application is wide-spread in industry as well as in academia. Since the early 1970s, when the first stable insect cell lines were established and the infectivity of bacu-lovirus in an in vitro culture system was demonstrated (1, 2), virtually thousands of reports have been published on the successful expression of proteins using this system as well as on method improvement. However, despite its popularity the system is labor intensive and time consuming. Moreover, adaptation of the system to multi-parallel (high-throughput) expression is much more difficult to achieve than with E. coli due to its far more complex nature. However, recent years have seen the development of strategies that have greatly enhanced the stream-lining and speed of baculovirus protein expression for increased throughput via use of automation and miniaturization. This chapter therefore tries to collate these developments in a series of protocols (which are modifications to standard procedure plus several new approaches) that will allow the user to expedite the speed and throughput of baculovirus-mediated protein expression and facilitate true multi-parallel, high-throughput protein expression profiling in insect cells. In addition we also provide a series of optimized protocols for small and large-scale transient insect cell expression that allow for both the rapid analysis of multiple constructs and the concomitant scale-up of those selected for on-going analysis. Since this approach is independent of viral propagation, the timelines for this approach are markedly shorter and offer a significant advantage over standard bacu-lovirus expression approach strategies in the context of HT applications.

  6. Gene expression in papillary thyroid carcinoma reveals highly consistent profiles

    PubMed Central

    Huang, Ying; Prasad, Manju; Lemon, William J.; Hampel, Heather; Wright, Fred A.; Kornacker, Karl; LiVolsi, Virginia; Frankel, Wendy; Kloos, Richard T.; Eng, Charis; Pellegata, Natalia S.; de la Chapelle, Albert

    2001-01-01

    Papillary thyroid carcinoma (PTC) is clinically heterogeneous. Apart from an association with ionizing radiation, the etiology and molecular biology of PTC is poorly understood. We used oligo-based DNA arrays to study the expression profiles of eight matched pairs of normal thyroid and PTC tissues. Additional PTC tumors and other tissues were studied by reverse transcriptase–PCR and immunohistochemistry. The PTCs showed concordant expression of many genes and distinct clustered profiles. Genes with increased expression in PTC included many encoding adhesion and extracellular matrix proteins. Expression was increased in 8/8 tumors for 24 genes and in 7/8 tumors for 22 genes. Among these genes were several previously known to be overexpressed in PTC, such as MET, LGALS3, KRT19, DPP4, MDK, TIMP1, and FN1. The numerous additional genes include CITED1, CHI3L1, ODZ1, N33, SFTPB, and SCEL. Reverse transcriptase–PCR showed high expression of CITED1, CHI3L1, ODZ1, and SCEL in 6/6 additional PTCs. Immunohistochemical analysis detected CITED1 and SFTPB in 49/52 and 39/52 PTCs, respectively, but not in follicular thyroid carcinoma and normal thyroid tissue. Genes underexpressed in PTC included tumor suppressors, thyroid function-related proteins, and fatty acid binding proteins. Expression was decreased in 7/8 tumors for eight genes and decreased in 6/8 tumors for 19 genes. We conclude that, despite its clinical heterogeneity, PTC is characterized by consistent and specific molecular changes. These findings reveal clues to the molecular pathways involved in PTC and may provide biomarkers for clinical use. PMID:11752453

  7. Shades of Emotion: What the Addition of Sunglasses or Masks to Faces Reveals about the Development of Facial Expression Processing

    ERIC Educational Resources Information Center

    Roberson, Debi; Kikutani, Mariko; Doge, Paula; Whitaker, Lydia; Majid, Asifa

    2012-01-01

    Three studies investigated developmental changes in facial expression processing, between 3 years-of-age and adulthood. For adults and older children, the addition of sunglasses to upright faces caused an equivalent decrement in performance to face inversion. However, younger children showed "better" classification of expressions of faces wearing…

  8. Thinking-for-Speaking of Chinese EFL Learners in the Use of Additional Expressions of Manner in the Expression of Motion Events

    ERIC Educational Resources Information Center

    Ziyan, Xu

    2013-01-01

    This study adopts Talmy's (1985, 1991, 2000) theory of linguistic typology and Slobin's (2003, 2004) hypothesis of Thinking-for-Speaking to investigate the use of additional expressions of Manner in the expression of Motion events by Chinese EFL learners in comparison with French EFL learners. The aim is to find out the Thinking-for-Speaking…

  9. Gene expression profiling in peanut using high density oligonucleotide microarrays

    PubMed Central

    Payton, Paxton; Kottapalli, Kameswara Rao; Rowland, Diane; Faircloth, Wilson; Guo, Baozhu; Burow, Mark; Puppala, Naveen; Gallo, Maria

    2009-01-01

    Background Transcriptome expression analysis in peanut to date has been limited to a relatively small set of genes and only recently has a significant number of ESTs been released into the public domain. Utilization of these ESTs for oligonucleotide microarrays provides a means to investigate large-scale transcript responses to a variety of developmental and environmental signals, ultimately improving our understanding of plant biology. Results We have developed a high-density oligonucleotide microarray for peanut using 49,205 publicly available ESTs and tested the utility of this array for expression profiling in a variety of peanut tissues. To identify putatively tissue-specific genes and demonstrate the utility of this array for expression profiling in a variety of peanut tissues, we compared transcript levels in pod, peg, leaf, stem, and root tissues. Results from this experiment showed 108 putatively pod-specific/abundant genes, as well as transcripts whose expression was low or undetected in pod compared to peg, leaf, stem, or root. The transcripts significantly over-represented in pod include genes responsible for seed storage proteins and desiccation (e.g., late-embryogenesis abundant proteins, aquaporins, legumin B), oil production, and cellular defense. Additionally, almost half of the pod-abundant genes represent unknown genes allowing for the possibility of associating putative function to these previously uncharacterized genes. Conclusion The peanut oligonucleotide array represents the majority of publicly available peanut ESTs and can be used as a tool for expression profiling studies in diverse tissues. PMID:19523230

  10. Modulation of GABA receptors expressed in Xenopus oocytes by 13-L-hydroxylinoleic acid and food additives.

    PubMed

    Aoshima, H; Tenpaku, Y

    1997-12-01

    To study the effects of 13-L-hydroxylinoleic acid (LOH) and food additives on gamma-aminobutyric acid (GABA) receptors, ionotropic GABA receptors were expressed in Xenopus oocytes by injecting mRNAs prepared from rat whole brain. LOH, which was prepared by reduction of 13-L-hydroperoxylinoleic acid (LOOH), inhibited the response of GABA receptors in the presence of high concentrations of GABA. LOH also inhibited nicotinic acetylcholine, glycine, and kainate receptors, while it had little effect on NMDA receptors expressed in Xenopus oocytes. However, LOH potentiated the response of GABA receptors as well as LOOH in the presence of low concentrations of GABA, possibly increasing the affinity of GABA for the receptors, while linoleic acid did not. Since some modification of the compounds seemed to change their effects on GABA receptors, the responses of GABA receptors elicited by 10 microM GABA were measured in the presence of compounds with various kinds of functional groups or the structural isomers of pentanol. Potentiation of GABA receptors depended strongly on the species of functional groups and also depended on the structure of the isomers. Then effects of various kinds of food additives on GABA receptors were also examined; perfumes such as alcohols or esters potentiated the responses strongly, while hexylamine, nicotinamide, or caffeine inhibited the responses, mainly in a competitive manner, and vanillin inhibited the responses noncompetitively. These results suggest the possibility that production of LOOH and LOH, or intake of much of some food additives, modulates the neural transmission in the brain, especially through ionotropic GABA receptors and changes the frame of the human mind, as alcohol or tobacco does.

  11. Genome-wide meta-analysis of maize heterosis reveals the potential role of additive gene expression at pericentromeric loci

    PubMed Central

    2014-01-01

    QTL, and also suggests a model for the potential role of additive expression in the formation and conservation of heterosis for GY via dominant, multigenic quantitative trait loci. Our findings contribute to a deeper understanding of the multifactorial phenomenon of heterosis, and thus to the breeding of new high yielding varieties. PMID:24693880

  12. Expressed Peptide Tags: An additional layer of data for genome annotation

    SciTech Connect

    Savidor, Alon; Donahoo, Ryan S; Hurtado-Gonzales, Oscar; Verberkmoes, Nathan C; Shah, Manesh B; Lamour, Kurt H; McDonald, W Hayes

    2006-01-01

    While genome sequencing is becoming ever more routine, genome annotation remains a challenging process. Identification of the coding sequences within the genomic milieu presents a tremendous challenge, especially for eukaryotes with their complex gene architectures. Here we present a method to assist the annotation process through the use of proteomic data and bioinformatics. Mass spectra of digested protein preparations of the organism of interest were acquired and searched against a protein database created by a six frame translation of the genome. The identified peptides were mapped back to the genome, compared to the current annotation, and then categorized as supporting or extending the current genome annotation. We named the classified peptides Expressed Peptide Tags (EPTs). The well annotated bacterium Rhodopseudomonas palustris was used as a control for the method and showed high degree of correlation between EPT mapping and the current annotation, with 86% of the EPTs confirming existing gene calls and less than 1% of the EPTs expanding on the current annotation. The eukaryotic plant pathogens Phytophthora ramorum and Phytophthora sojae, whose genomes have been recently sequenced and are much less well annotated, were also subjected to this method. A series of algorithmic steps were taken to increase the confidence of EPT identification for these organisms, including generation of smaller sub-databases to be searched against, and definition of EPT criteria that accommodates the more complex eukaryotic gene architecture. As expected, the analysis of the Phytophthora species showed less correlation between EPT mapping and their current annotation. While ~77% of Phytophthora EPTs supported the current annotation, a portion of them (7.2% and 12.6% for P. ramorum and P. sojae, respectively) suggested modification to current gene calls or identified novel genes that were missed by the current genome annotation of these organisms.

  13. 49 CFR 236.1007 - Additional requirements for high-speed service.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 49 Transportation 4 2012-10-01 2012-10-01 false Additional requirements for high-speed service..., AND APPLIANCES Positive Train Control Systems § 236.1007 Additional requirements for high-speed... by this subpart, and which have been utilized on high-speed rail systems with similar technical...

  14. 49 CFR 236.1007 - Additional requirements for high-speed service.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 49 Transportation 4 2014-10-01 2014-10-01 false Additional requirements for high-speed service..., AND APPLIANCES Positive Train Control Systems § 236.1007 Additional requirements for high-speed... by this subpart, and which have been utilized on high-speed rail systems with similar technical...

  15. 49 CFR 236.1007 - Additional requirements for high-speed service.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 49 Transportation 4 2010-10-01 2010-10-01 false Additional requirements for high-speed service..., AND APPLIANCES Positive Train Control Systems § 236.1007 Additional requirements for high-speed... by this subpart, and which have been utilized on high-speed rail systems with similar technical...

  16. 49 CFR 236.1007 - Additional requirements for high-speed service.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 49 Transportation 4 2013-10-01 2013-10-01 false Additional requirements for high-speed service..., AND APPLIANCES Positive Train Control Systems § 236.1007 Additional requirements for high-speed... by this subpart, and which have been utilized on high-speed rail systems with similar technical...

  17. 49 CFR 236.1007 - Additional requirements for high-speed service.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 49 Transportation 4 2011-10-01 2011-10-01 false Additional requirements for high-speed service..., AND APPLIANCES Positive Train Control Systems § 236.1007 Additional requirements for high-speed... by this subpart, and which have been utilized on high-speed rail systems with similar technical...

  18. Baculovirus expression system and method for high throughput expression of genetic material

    DOEpatents

    Clark, Robin; Davies, Anthony

    2001-01-01

    The present invention provides novel recombinant baculovirus expression systems for expressing foreign genetic material in a host cell. Such expression systems are readily adapted to an automated method for expression foreign genetic material in a high throughput manner. In other aspects, the present invention features a novel automated method for determining the function of foreign genetic material by transfecting the same into a host by way of the recombinant baculovirus expression systems according to the present invention.

  19. [Highly efficient and rapid capillary electrophoretic analysis of seven organic acid additives in beverages using polymeric ionic liquid as additive].

    PubMed

    Han, Haifeng; Wang, Qing; Liu, Xi; Jiang, Shengxiang

    2012-05-01

    A new capillary electrophoretic method for the rapid and direct separation of seven organic acids in beverages was developed, with poly (1-vinyl-3-butylimidazolium bromide) as the reliable background electrolyte modifier to reverse the direction of anode electroosmotic flow (EOF) severely. Several factors that affected the separation efficiency were investigated in detail. The optimal running buffer consisted of 125 mmol/L sodium dihydrogen phosphate (pH 6.5) and 0.01 g/L poly (1-vinyl-3-butylimidazolium bromide). Highly efficient separation (105,000 to 636,000 plates/m) was achieved within 4 min and standard deviations of the migration times (n=3) were lower than 0.0213 min under optimal conditions. The limits of detection (S/N = 3) ranged from 0.001 to 0.05 g/L. The present method was applied to determine a beverage sample (Mirinda) for sodium citrate, benzoic acid and sorbic acid with concentration of 2.64, 0.10 and 0.08 g/L, respectively. The recoveries of the three analytes in the sample were 100.3%, 100.7% and 131.7%, respectively. The method is simple, rapid, inexpensive, and can be applied to determine organic acids as additives in beverages.

  20. Sequence and Expression Analyses of Ethylene Response Factors Highly Expressed in Latex Cells from Hevea brasiliensis

    PubMed Central

    Piyatrakul, Piyanuch; Yang, Meng; Putranto, Riza-Arief; Pirrello, Julien; Dessailly, Florence; Hu, Songnian; Summo, Marilyne; Theeravatanasuk, Kannikar; Leclercq, Julie; Kuswanhadi; Montoro, Pascal

    2014-01-01

    The AP2/ERF superfamily encodes transcription factors that play a key role in plant development and responses to abiotic and biotic stress. In Hevea brasiliensis, ERF genes have been identified by RNA sequencing. This study set out to validate the number of HbERF genes, and identify ERF genes involved in the regulation of latex cell metabolism. A comprehensive Hevea transcriptome was improved using additional RNA reads from reproductive tissues. Newly assembled contigs were annotated in the Gene Ontology database and were assigned to 3 main categories. The AP2/ERF superfamily is the third most represented compared with other transcription factor families. A comparison with genomic scaffolds led to an estimation of 114 AP2/ERF genes and 1 soloist in Hevea brasiliensis. Based on a phylogenetic analysis, functions were predicted for 26 HbERF genes. A relative transcript abundance analysis was performed by real-time RT-PCR in various tissues. Transcripts of ERFs from group I and VIII were very abundant in all tissues while those of group VII were highly accumulated in latex cells. Seven of the thirty-five ERF expression marker genes were highly expressed in latex. Subcellular localization and transactivation analyses suggested that HbERF-VII candidate genes encoded functional transcription factors. PMID:24971876

  1. Differential Expression of Functional Fc-Receptors and Additional Immune Complex Receptors on Mouse Kidney Cells

    PubMed Central

    Suwanichkul, Adisak; Wenderfer, Scott E.

    2013-01-01

    The precise mechanisms by which circulating immune complexes accumulate in the kidney to form deposits in glomerulonephritis are not well understood. In particular, the role of resident cells within glomeruli of the kidney has been widely debated. Immune complexes have been shown to bind one glomerular cell type (mesangial cells) leading to functional responses such as pro-inflammatory cytokine production. To further assess the presence of functional immunoreceptors on resident glomerular cells, cultured mouse renal epithelial, endothelial, and mesangial cells were treated with heat-aggregated mouse IgG or preformed murine immune complexes. Mesangial and renal endothelial cells were found to bind IgG complexes, whereas glomerular epithelial cell binding was minimal. A blocking antibody for Fc-gamma receptors reduced binding to mesangial cells but not renal endothelial cells, suggesting differential immunoreceptor utilization. RT-PCR and immunostaining based screening of cultured renal endothelial cells showed limited low-level expression of known Fc-receptors and Igbinding proteins. The interaction between mesangial cells and renal endothelial cells and immune complexes resulted in distinct, cell-specific patterns of chemokine and cytokine production. This novel pathway involving renal endothelial cells likely contributes to the predilection of circulating immune complex accumulation within the kidney and to the inflammatory responses that drive kidney injury. PMID:23911392

  2. Molecular cloning and expression of an additional epidermal growth factor receptor-related gene.

    PubMed Central

    Plowman, G D; Whitney, G S; Neubauer, M G; Green, J M; McDonald, V L; Todaro, G J; Shoyab, M

    1990-01-01

    Epidermal growth factor (EGF), transforming growth factor alpha (TGF-alpha), and amphiregulin are structurally and functionally related growth regulatory proteins. These secreted polypeptides all bind to the 170-kDa cell-surface EGF receptor, activating its intrinsic kinase activity. However, amphiregulin exhibits different activities than EGF and TGF-alpha in a number of biological assays. Amphiregulin only partially competes with EGF for binding EGF receptor, and amphiregulin does not induce anchorage-independent growth of normal rat kidney cells (NRK) in the presence of TGF-beta. Amphiregulin also appears to abrogate the stimulatory effect of TGF-alpha on the growth of several aggressive epithelial carcinomas that overexpress EGF receptor. These findings suggest that amphiregulin may interact with a separate receptor in certain cell types. Here we report the cloning of another member of the human EGF receptor (HER) family of receptor tyrosine kinases, which we have named "HER3/ERRB3." The cDNA was isolated from a human carcinoma cell line, and its 6-kilobase transcript was identified in various human tissues. We have generated peptide-specific antisera that recognizes the 160-kDa HER3 protein when transiently expressed in COS cells. These reagents will allow us to determine whether HER3 binds amphiregulin or other growth regulatory proteins and what role HER3 protein plays in the regulation of cell growth. Images PMID:2164210

  3. The addition of high magnifying endoscopy improves rates of high confidence optical diagnosis of colorectal polyps

    PubMed Central

    Iwatate, Mineo; Sano, Yasushi; Hattori, Santa; Sano, Wataru; Hasuike, Noriaki; Ikumoto, Taro; Kotaka, Masahito; Murakami, Yoshitaka; Hewett, David G.; Soetikno, Roy; Kaltenbach, Tonya; Fujimori, Takahiro

    2015-01-01

    Background and study aims: The real-time optical diagnosis of colorectal polyps with high confidence predictions can achieve high levels of accuracy. Increasing the rates of high confidence optical diagnosis can improve the clinical application of real-time optical diagnosis in routine practice. The primary aim of this prospective study was to evaluate whether high magnifying endoscopy improves the rates of high confidence narrow-band imaging (NBI) – based optical diagnosis for differentiating between neoplastic and non-neoplastic colorectal lesions according to the NBI international colorectal endoscopic (NICE) classification. Patients and methods: Consecutive adult patients undergoing colonoscopy with a high magnifying (maximum, × 80) colonoscope between April and August 2012 were recruited. The optical diagnosis for each polyp was evaluated during colonoscopy in two consecutive stages by the same endoscopist, who first used NBI with non-magnifying endoscopy (NBI-NME), then NBI with magnifying endoscopy (NBI-ME). A level of confidence was assigned to each prediction. Results: The analysis included 124 patients (mean age, 56.4 years; male-to-female ratio, 72:52) with 248 polyps smaller than 10 mm. Of the 248 polyps, 210 were 1 to 5 mm in size and 38 were 6 to 9 mm in size; 77 polyps were hyperplastic, 4 were sessile serrated adenomas/polyps, 160 were low grade adenomas, 5 were high grade adenomas, and 2 were deep submucosal invasive carcinomas. The rate of high confidence optical diagnosis when NBI-ME was used was significantly higher than the rate when NBI-NME was used for diminutive (1 – 5 mm) polyps (92.9 % vs 79.5 %, P < 0.001) and for small (6 – 9 mm) polyps (94.7 % vs 84.2 %, P = 0.048). Conclusion: High magnifying endoscopy significantly improved the rates of high confidence NBI-based optical diagnosis of diminutive and small colorectal polyps. Study registration: UMIN 000007608 PMID:26135657

  4. High Energy Density Additives for Hybrid Fuel Rockets to Improve Performance and Enhance Safety

    NASA Technical Reports Server (NTRS)

    Jaffe, Richard L.

    2014-01-01

    We propose a conceptual study of prototype strained hydrocarbon molecules as high energy density additives for hybrid rocket fuels to boost the performance of these rockets without compromising safety and reliability. Use of these additives could extend the range of applications for which hybrid rockets become an attractive alternative to conventional solid or liquid fuel rockets. The objectives of the study were to confirm and quantify the high enthalpy of these strained molecules and to assess improvement in rocket performance that would be expected if these additives were blended with conventional fuels. We confirmed the chemical properties (including enthalpy) of these additives. However, the predicted improvement in rocket performance was too small to make this a useful strategy for boosting hybrid rocket performance.

  5. Effects of maternally exposed coloring food additives on receptor expressions related to learning and memory in rats.

    PubMed

    Ceyhan, Betul Mermi; Gultekin, Fatih; Doguc, Duygu Kumbul; Kulac, Esin

    2013-06-01

    Exposure to artificial food colors and additives (AFCAs) has been implicated in the induction and severity of some childhood behavioral and learning disabilities. N-methyl-D-aspartate receptors (NMDARs) and nicotinic acetylcholine receptors (nACHRs) are thought to be effective in the learning and memory-generating process. In this study, we investigated the effects of intrauterine exposure to AFCAs on subunit concentrations of NMDARs and nAChRs isoforms in rats. We administered a mixture of AFCAs (Eritrosin, Ponceau 4R, Allura Red AC, Sunset Yellow FCF, Tartrazin, Amaranth, Brilliant Blue, Azorubin and Indigotin) to female rats before and during gestation. The concentration of NR2A and NR2B subunits and nAChR α7, α4β2 isoforms in their offspring's hippocampi were measured by Western Blotting. Expressions of NR2B and nAChR β2 were significantly increased (17% and 6.70%, respectively), whereas expression of nAChR α4 was significantly decreased (5.67%) in male experimental group compared to the male control group (p<0.05). In the female experimental group, AFCAs caused a 14% decrease in NR2B expression when compared to the female control group (p<0.05). Our results indicate that exposure to AFCAs during the fetal period may lead to alterations in expressions of NMDARs and nAChRs in adulthood. These alterations were different between male and female genders. PMID:23429044

  6. Highly Diastereoselective Chelation-controlled Additions to α-Silyloxy Ketones

    PubMed Central

    Stanton, Gretchen R.; Koz, Gamze

    2011-01-01

    The polar Felkin-Anh, Cornforth, and Cram-chelation models predict that the addition of organometallic reagents to silyl–protected α–hydroxy ketones proceeds via a non-chelation pathway to give anti-diol addition products. This prediction has held true for the vast majority of additions reported in the literature and few methods for chelation-controlled additions of organometallic reagents to silyl–protected α–hydroxy ketones have been introduced. Herein, we present a general and highly diastereoselective method for the addition of dialkylzincs and (E)-di-, (E)-tri- and (Z)-disubstituted vinylzinc reagents to α-silyloxy ketones using alkyl zinc halide Lewis acids, RZnX, to give chelation-controlled products (dr ≥18:1). The compatibility of organozinc reagents with other functional groups makes this method potentially very useful in complex molecule synthesis. PMID:21534530

  7. Novel high refractive index, thermally conductive additives for high brightness white LEDs

    NASA Astrophysics Data System (ADS)

    Hutchison, Richard Stephen

    In prior works the inclusion of nanoparticle fillers has typically been shown to increase the thermal conductivity or refractive index of polymer nanocomposites separately. High refractive index zirconia nanoparticles have already proved their merit in increasing the optical efficiency of encapsulated light emitting diodes. However, the thermal properties of zirconia-silicone nanocomposites have yet to be investigated. While phosphor-converted light emitting diodes are at the forefront of solid-state lighting technologies for producing white light, they are plagued by efficiency losses due to excessive heating at the semiconductor die and in and around the phosphor particles, as well as photon scattering losses in the phosphor layer. It would then be of great interest if the high refractive index nanoparticles were found to both be capable of increasing the refractive index, thus reducing the optical scattering, and also the thermal conductivity, channeling more heat away from the LED die and phosphors, mitigating efficiency losses from heat. Thermal conductance measurements on unfilled and nanoparticle loaded silicone samples were conducted to quantify the effect of the zirconia nanoparticle loading on silicone nanocomposite thermal conductivity. An increase in thermal conductivity from 0.27 W/mK to 0.49 W/mK from base silicone to silicone with 33.5 wt% zirconia nanoparticles was observed. This trend closely mirrored a basic rule of mixtures prediction, implying a further enhancement in thermal conductivity could be achieved at higher nanoparticle loadings. The optical properties of transparency and light extraction efficiency of these composites were also investigated. While overall the zirconia nanocomposite showed good transparency, there was a slight decrease at the shorter wavelengths with increasing zirconia content. For longer wavelength LEDs, such as green or red, this might not matter, but phosphor-converted white LEDs use a blue LED as the photon source

  8. MRP1 and P-glycoprotein expression assays would be useful in the additional detection of treatment non-responders in CML patients without ABL1 mutation.

    PubMed

    Park, Sang Hyuk; Park, Chan-Jeoung; Kim, Dae-Young; Lee, Bo-Ra; Kim, Young Jin; Cho, Young-Uk; Jang, Seongsoo

    2015-10-01

    We evaluated the ability of the rhodamine-123 efflux assay, multidrug resistance-associated protein-1 (MRP1) expression assay and P-glycoprotein (Pgp) expression assay to discriminate chronic myelogenous leukemia (CML) patients who had failed treatment or were at risk of failure. Each assay was performed in blood samples from CML patients (n=224) treated with tyrosine kinase inhibitors, taken at diagnosis (n=14) and follow-up (n=210). Patient samples were categorized as optimal response (n=120), suboptimal response (n=54), and treatment failure (n=36). Treatment-failed patients had a significantly higher MRP1 expression (5.24% vs. 3.54%, P=0.006) and Pgp expression (5.25% vs. 3.48%, P=0.005) than responders. Both MRP1 (%) and Pgp (%) were highly specific (95.2% and 94.5%) and relatively accurate (83.0% and 82.5%) in the detection of treatment non-responders. Of treatment-failed patients, 41.2% had a positive result in at least one assay and of these patients without ABL1 kinase domain mutation, 51.9% were positive in at least one assay. However, the rhodamine-123 efflux assay failed to discriminate two patient groups. Thus, both MRP1 and Pgp expression assays could be useful for additional identification of treatment non-responders in CML patients without ABL1 mutation.

  9. Highly Enantioselective Rhodium-Catalyzed Addition of Arylboroxines to Simple Aryl Ketones: Efficient Synthesis of Escitalopram.

    PubMed

    Huang, Linwei; Zhu, Jinbin; Jiao, Guangjun; Wang, Zheng; Yu, Xingxin; Deng, Wei-Ping; Tang, Wenjun

    2016-03-24

    Highly enantioselective additions of arylboroxines to simple aryl ketones have been achieved for the first time with a Rh/(R,R,R,R)-WingPhos catalyst, thus providing a range of chiral diaryl alkyl carbinols with excellent ee values and yields. (R,R,R,R)-WingPhos has been proven to be crucial for the high reactivity and enantioselectivity. The method has enabled a new, concise, and enantioselective synthesis of the antidepressant drug escitalopram. PMID:26933831

  10. How to achieve high-level expression of microbial enzymes

    PubMed Central

    Liu, Long; Yang, Haiquan; Shin, Hyun-dong; Chen, Rachel R.; Li, Jianghua; Du, Guocheng; Chen, Jian

    2013-01-01

    Microbial enzymes have been used in a large number of fields, such as chemical, agricultural and biopharmaceutical industries. The enzyme production rate and yield are the main factors to consider when choosing the appropriate expression system for the production of recombinant proteins. Recombinant enzymes have been expressed in bacteria (e.g., Escherichia coli, Bacillus and lactic acid bacteria), filamentous fungi (e.g., Aspergillus) and yeasts (e.g., Pichia pastoris). The favorable and very advantageous characteristics of these species have resulted in an increasing number of biotechnological applications. Bacterial hosts (e.g., E. coli) can be used to quickly and easily overexpress recombinant enzymes; however, bacterial systems cannot express very large proteins and proteins that require post-translational modifications. The main bacterial expression hosts, with the exception of lactic acid bacteria and filamentous fungi, can produce several toxins which are not compatible with the expression of recombinant enzymes in food and drugs. However, due to the multiplicity of the physiological impacts arising from high-level expression of genes encoding the enzymes and expression hosts, the goal of overproduction can hardly be achieved, and therefore, the yield of recombinant enzymes is limited. In this review, the recent strategies used for the high-level expression of microbial enzymes in the hosts mentioned above are summarized and the prospects are also discussed. We hope this review will contribute to the development of the enzyme-related research field. PMID:23686280

  11. HEAT: High accuracy extrapolated ab initio thermochemistry. III. Additional improvements and overview.

    SciTech Connect

    Harding, M. E.; Vazquez, J.; Ruscic, B.; Wilson, A. K.; Gauss, J.; Stanton, J. F.; Chemical Sciences and Engineering Division; Univ. t Mainz; The Univ. of Texas; Univ. of North Texas

    2008-01-01

    Effects of increased basis-set size as well as a correlated treatment of the diagonal Born-Oppenheimer approximation are studied within the context of the high-accuracy extrapolated ab initio thermochemistry (HEAT) theoretical model chemistry. It is found that the addition of these ostensible improvements does little to increase the overall accuracy of HEAT for the determination of molecular atomization energies. Fortuitous cancellation of high-level effects is shown to give the overall HEAT strategy an accuracy that is, in fact, higher than most of its individual components. In addition, the issue of core-valence electron correlation separation is explored; it is found that approximate additive treatments of the two effects have limitations that are significant in the realm of <1 kJ mol{sup -1} theoretical thermochemistry.

  12. Radiative properties of ceramic metal-halide high intensity discharge lamps containing additives in argon plasma

    NASA Astrophysics Data System (ADS)

    Cressault, Yann; Teulet, Philippe; Zissis, Georges

    2016-07-01

    The lighting represents a consumption of about 19% of the world electricity production. We are thus searching new effective and environment-friendlier light sources. The ceramic metal-halide high intensity lamps (C-MHL) are one of the options for illuminating very high area. The new C-MHL lamps contain additives species that reduce mercury inside and lead to a richer spectrum in specific spectral intervals, a better colour temperature or colour rendering index. This work is particularly focused on the power radiated by these lamps, estimated using the net emission coefficient, and depending on several additives (calcium, sodium, tungsten, dysprosium, and thallium or strontium iodides). The results show the strong influence of the additives on the power radiated despite of their small quantity in the mixtures and the increase of visible radiation portion in presence of dysprosium.

  13. Production of fired construction brick from high sulfate-containing fly ash with boric acid addition.

    PubMed

    Başpinar, M Serhat; Kahraman, Erhan; Görhan, Gökhan; Demir, Ismail

    2010-01-01

    The increase of power plant capacity has led to the production of an increasing amount of fly ash that causes high environmental impact in Turkey. Some of the fly ash is utilized within the fired brick industry but high sulfate-containing fly ash creates severe problems during sintering of the fired brick. This study attempted to investigate the potential for converting high sulfate-containing fly ash into useful material for the construction industry by the addition of boric acid. The chemical and mineralogical composition of fly ash and clay were investigated. Boric acid (H(3)BO(3)) was added to fly ash-clay mixtures with up to 5 wt.%. Six different series of test samples were produced by uniaxial pressing. The samples were fired at the industrial clay-brick firing temperatures of 800, 900 and 1000 degrees C. The microstructures of the fired samples were investigated by scanning electron microscopy and some physical and mechanical properties were measured. It was concluded that the firing at conventional brick firing temperature of high sulfate fly ash without any addition of boric acid resulted in very weak strength bricks. The addition of boric acid and clay simultaneously to the high sulfate- containing fly ash brick dramatically increased the compressive strength of the samples at a firing temperature of 1000 degrees C by modifying the sintering behaviour of high sulfate fly ash.

  14. Production of fired construction brick from high sulfate-containing fly ash with boric acid addition.

    PubMed

    Başpinar, M Serhat; Kahraman, Erhan; Görhan, Gökhan; Demir, Ismail

    2010-01-01

    The increase of power plant capacity has led to the production of an increasing amount of fly ash that causes high environmental impact in Turkey. Some of the fly ash is utilized within the fired brick industry but high sulfate-containing fly ash creates severe problems during sintering of the fired brick. This study attempted to investigate the potential for converting high sulfate-containing fly ash into useful material for the construction industry by the addition of boric acid. The chemical and mineralogical composition of fly ash and clay were investigated. Boric acid (H(3)BO(3)) was added to fly ash-clay mixtures with up to 5 wt.%. Six different series of test samples were produced by uniaxial pressing. The samples were fired at the industrial clay-brick firing temperatures of 800, 900 and 1000 degrees C. The microstructures of the fired samples were investigated by scanning electron microscopy and some physical and mechanical properties were measured. It was concluded that the firing at conventional brick firing temperature of high sulfate fly ash without any addition of boric acid resulted in very weak strength bricks. The addition of boric acid and clay simultaneously to the high sulfate- containing fly ash brick dramatically increased the compressive strength of the samples at a firing temperature of 1000 degrees C by modifying the sintering behaviour of high sulfate fly ash. PMID:19423597

  15. Midrapidity inclusive densities in high energy pp collisions in additive quark model

    NASA Astrophysics Data System (ADS)

    Shabelski, Yu. M.; Shuvaev, A. G.

    2016-08-01

    High energy (CERN SPS and LHC) inelastic pp (pbar{p}) scattering is treated in the framework of the additive quark model together with Pomeron exchange theory. We extract the midrapidity inclusive density of the charged secondaries produced in a single quark-quark collision and investigate its energy dependence. Predictions for the π p collisions are presented.

  16. The Modern U.S. High School Astronomy Course, Its Status and Makeup II: Additional Results

    ERIC Educational Resources Information Center

    Krumenaker, Larry

    2009-01-01

    A postal survey of high school astronomy teachers strongly confirms many results of an earlier electronic survey. Additional and new results include a measure of the level of inquiry (more structured inquiry and teacher-led) in the classroom as well as data showing that more emphasis is given to traditional topics than to contemporary astronomy…

  17. Brief Report: Additive and Subtractive Counterfactual Reasoning of Children with High-Functioning Autism Spectrum Disorders

    ERIC Educational Resources Information Center

    Begeer, Sander; Terwogt, Mark Meerum; Lunenburg, Patty; Stegge, Hedy

    2009-01-01

    The development of additive ("If only I had done...") and subtractive ("If only I had not done....") counterfactual reasoning was examined in children with High Functioning Autism Spectrum Disorders (HFASD) (n = 72) and typically developing controls (n = 71), aged 6-12 years. Children were presented four stories where they could generate…

  18. Effect of additives on the properties of polyaniline nanofibers prepared by high gravity chemical oxidative polymerization.

    PubMed

    Zhao, Yibo; Arowo, Moses; Wu, Wei; Chen, Jianfeng

    2015-05-12

    Polyaniline (PANI) nanofibers with improved properties were prepared by high gravity chemical oxidative polymerization in a rotating packed bed with the assistance of p-aminodiphenylamine (AD) and p-phenylenediamine (AP). The effects of reactor type, additive dosage, reaction temperature, and high-gravity level on the properties of products were investigated in detail. Three conclusions were made: (1) a small amount of additive can significantly improve some properties of the nanofibers such as uniformity, specific surface area, and specific capacitance; (2) in order to obtain high-quality nanofibers, the high-gravity level should coordinate with the reaction rate; (3) the molecular weight and conductivity of PANI decrease with the increase of additive dosage. The products have larger specific surface areas of up to 73.9 and 68.4 m(2)/g and consequently improved specific capacitance of up to 527.5 and 552 F/g for the PANI nanofibers prepared with AD and AP, respectively. However, the specific surface area and specific capacitance of pure PANI are only 49.1 m(2)/g and 333.3 F/g, respectively. This research provides a simple, reliable, and scalable method to produce PANI nanofibers of high performances.

  19. Combinatorial therapeutic activation with heparin and AICAR stimulates additive effects on utrophin A expression in dystrophic muscles.

    PubMed

    Péladeau, Christine; Ahmed, Aatika; Amirouche, Adel; Crawford Parks, Tara E; Bronicki, Lucas M; Ljubicic, Vladimir; Renaud, Jean-Marc; Jasmin, Bernard J

    2016-01-01

    Upregulation of utrophin A is an attractive therapeutic strategy for treating Duchenne muscular dystrophy (DMD). Over the years, several studies revealed that utrophin A is regulated by multiple transcriptional and post-transcriptional mechanisms, and that pharmacological modulation of these pathways stimulates utrophin A expression in dystrophic muscle. In particular, we recently showed that activation of p38 signaling causes an increase in the levels of utrophin A mRNAs and protein by decreasing the functional availability of the destabilizing RNA-binding protein called K-homology splicing regulatory protein, thereby resulting in increases in the stability of existing mRNAs. Here, we treated 6-week-old mdx mice for 4 weeks with the clinically used anticoagulant drug heparin known to activate p38 mitogen-activated protein kinase, and determined the impact of this pharmacological intervention on the dystrophic phenotype. Our results show that heparin treatment of mdx mice caused a significant ∼1.5- to 3-fold increase in utrophin A expression in diaphragm, extensor digitorum longus and tibialis anterior (TA) muscles. In agreement with these findings, heparin-treated diaphragm and TA muscle fibers showed an accumulation of utrophin A and β-dystroglycan along their sarcolemma and displayed improved morphology and structural integrity. Moreover, combinatorial drug treatment using both heparin and 5-amino-4-imidazolecarboxamide riboside (AICAR), the latter targeting 5' adenosine monophosphate-activated protein kinase and the transcriptional activation of utrophin A, caused an additive effect on utrophin A expression in dystrophic muscle. These findings establish that heparin is a relevant therapeutic agent for treating DMD, and illustrate that combinatorial treatment of heparin with AICAR may serve as an effective strategy to further increase utrophin A expression in dystrophic muscle via activation of distinct signaling pathways.

  20. Immunoinformatics study on highly expressed Mycobacterium tuberculosis genes during infection.

    PubMed

    Nguyen Thi, Le Thuy; Sarmiento, Maria Elena; Calero, Romel; Camacho, Frank; Reyes, Fatima; Hossain, Md Murad; Gonzalez, Gustavo Sierra; Norazmi, Mohd Nor; Acosta, Armando

    2014-09-01

    The most important targets for vaccine development are the proteins that are highly expressed by the microorganisms during infection in-vivo. A number of Mycobacterium tuberculosis (Mtb) proteins are also reported to be expressed in-vivo at different phases of infection. In the present study, we analyzed multiple published databases of gene expression profiles of Mtb in-vivo at different phases of infection in animals and humans and selected 38 proteins that are highly expressed in the active, latent and reactivation phases. We predicted T- and B-cell epitopes from the selected proteins using HLAPred for T-cell epitope prediction and BCEPred combined with ABCPred for B-cell epitope prediction. For each selected proteins, regions containing both T- and B-cell epitopes were identified which might be considered as important candidates for vaccine design against tuberculosis.

  1. Additive Regulation of Adiponectin Expression by the Mediterranean Diet Olive Oil Components Oleic Acid and Hydroxytyrosol in Human Adipocytes

    PubMed Central

    Scoditti, Egeria; Massaro, Marika; Carluccio, Maria Annunziata; Pellegrino, Mariangela; Wabitsch, Martin; Calabriso, Nadia; Storelli, Carlo; De Caterina, Raffaele

    2015-01-01

    Adiponectin, an adipocyte-derived insulin-sensitizing and anti-inflammatory hormone, is suppressed in obesity through mechanisms involving chronic inflammation and oxidative stress. Olive oil consumption is associated with beneficial cardiometabolic actions, with possible contributions from the antioxidant phenol hydroxytyrosol (HT) and the monounsaturated fatty acid oleic acid (OA, 18:1n-9 cis), both possessing anti-inflammatory and vasculo-protective properties. We determined the effects of HT and OA, alone and in combination, on adiponectin expression in human and murine adipocytes under pro-inflammatory conditions induced by the cytokine tumor necrosis factor(TNF)-α. We used human Simpson-Golabi-Behmel syndrome (SGBS) adipocytes and murine 3T3-L1 adipocytes as cell model systems, and pretreated them with 1-100 μmol/L OA, 0.1-20 μmol/L HT or OA plus HT combination before stimulation with 10 ng/mL TNF-α. OA or HT significantly (P<0.05) prevented TNF-α-induced suppression of total adiponectin secretion (by 42% compared with TNF-α alone) as well as mRNA levels (by 30% compared with TNF-α alone). HT and OA also prevented—by 35%—TNF-α-induced downregulation of peroxisome proliferator-activated receptor PPARγ. Co-treatment with HT and OA restored adiponectin and PPARγ expression in an additive manner compared with single treatments. Exploring the activation of JNK, which is crucial for both adiponectin and PPARγ suppression by TNF-α, we found that HT and OA additively attenuated TNF-α-stimulated JNK phosphorylation (up to 55% inhibition). In conclusion, the virgin olive oil components OA and HT, at nutritionally relevant concentrations, have additive effects in preventing adiponectin downregulation in inflamed adipocytes through an attenuation of JNK-mediated PPARγ suppression. PMID:26030149

  2. High-Throughput Baculovirus Expression System for Membrane Protein Production.

    PubMed

    Kalathur, Ravi C; Panganiban, Marinela; Bruni, Renato

    2016-01-01

    The ease of use, robustness, cost-effectiveness, and posttranslational machinery make baculovirus expression system a popular choice for production of eukaryotic membrane proteins. This system can be readily adapted for high-throughput operations. This chapter outlines the techniques and procedures for cloning, transfection, small-scale production, and purification of membrane protein samples in a high-throughput manner. PMID:27485337

  3. High expression of DEK predicts poor prognosis of gastric adenocarcinoma

    PubMed Central

    2014-01-01

    Background DEK, as an oncoprotein, plays an important role in cancer development and progression. This study aimed to investigate the clinicopathological significance of DEK overexpression in patients with gastric cancer. Materials and methods The expression of DEK protein was evaluated by immunohistochemical (IHC) staining of 172 gastric cancer samples with complete clinicopathological features, and the correlation between DEK expression and clinicopathological features was examined. Survival rates were also calculated using the Kaplan-Meier method in gastric cancer patients with complete survival data. Results DEK protein showed a strictly nuclear staining pattern in gastric cancers with IHC and immunofluorescence. The strongly positive rate of DEK protein was 60.5% (104/172) in gastric cancers, which was significantly higher than that in either gastric dysplasia (19.4%, 7/36) or adjacent normal mucosa (0%, 0/27). DEK expression in gastric cancer correlated to tumor size, differentiation, clinical stage, disease-free survival, and overall survival rates. Further analysis showed that patients with early-stage gastric cancer and high DEK expression had shorter disease-free survival and overall survival duration than those with low DEK expression. Conclusion High level of DEK protein expression predicts the poor prognosis of patients with gastric cancer. DEK expression might be potentially used as an independent effective biomarker for prognostic evaluation of gastric cancers. Virtual slides The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/5050145571193097 PMID:24650035

  4. Properties and Microstructural Characteristic of Kaolin Geopolymer Ceramics with Addition of Ultra High Molecular Weight Polyethylene

    NASA Astrophysics Data System (ADS)

    Ahmad, Romisuhani; Bakri Abdullah, Mohd Mustafa Al; Hussin, Kamarudin; Sandu, Andrei Victor; Binhussain, Mohammed; Ain Jaya, Nur

    2016-06-01

    In this paper, the mechanical properties and microstructure of kaolin geopolymer ceramics with addition of Ultra High Molecular Weight Polyethylene were studied. Inorganic polymers based on alumina and silica polysialate units were synthesized at room temperature from kaolin and sodium silicate in a highly alkaline medium, followed by curing and drying at 80 °C. Alkaline activator was formed by mixing the 12 M NaOH solution with sodium silicate at a ratio of 0.24. Addition of Ultra High Molecular Weight Polyethylene to the kaolin geopolymer are fabricated with Ultra High Molecular Weight Polyethylene content of 2, 4, 6 and 8 (wt. %) by using powder metallurgy method. The samples were heated at 1200 °C and the strength and morphological were tested. It was found that the flexural strength for the kaolin geopolymer ceramics with addition of UHMWPE were improved and generally increased with the increasing of UHMWPE loading. The result revealed that the optimum flexural strength was obtained at UHMWPE loading of 4 wt. % (92.1 MPa) and the flexural strength started to decrease. Microstructural analysis showed the samples appeared to have more number of pores and connected of pores increased with the increasing of UHMWPE content.

  5. Fabrication and characterization of Si3N4 ceramics without additives by high pressure hot pressing

    NASA Technical Reports Server (NTRS)

    Shimada, M.; Tanaka, A.; Yamada, T.; Koizumi, M.

    1984-01-01

    High pressure hot-pressing of Si3N4 without additives was performed using various kinds of Si3N4 powder as starting materials, and the relation between densification and alpha-beta phase transformation was studied. The temperature dependences of Vickers microhardness and fracture toughness were also examined. Densification of Si3N4 was divided into three stages, and it was found that densification and phase transformation of Si3N4 under pressure were closely associated. The results of the temperature dependence of Vickers microhardness indicated that the high-temperature hardness was strongly influenced not only by the density and microstructure of sintered body but also by the purity of starting powder. The fracture toughness values of Si3N4 bodies without additives were 3.29-4.39 MN/m to the 3/2 power and independent of temperature up to 1400 C.

  6. The use of Electrolyte Additives to Improve the High Temperature Resilience of Li-Ion Cells

    NASA Technical Reports Server (NTRS)

    Smart, Marshall C.; Lucht, B. L.; Ratnakumar, Bugga V.

    2007-01-01

    This viewgraph presentation reviews the use of electrolyte additves to improve the resillience of Lithium ion cells. The objective of this work is to identify lithium-ion electrolytes, which will lead to Li-ion cells with a wide operational temperature range (+60 to -60 C), and to develop Li-ion electrolytes which result in cells that display improved high temperature resilience. Significant improvement in the high temperature resilience of Li-ion cells containing these additives was observed, with the most dramatic benefit being displayed by addition of DMAc. When the electrochemical properties of the individual electrodes were analyzed, the degradation of the anode kinetics was slowed most dramatically by the incorporation of DMAc into the electrolytes. Whereas, the greatest retention in the cathode kinetics was observed in the cell containing the electrolyte with VC added.

  7. Phosphorous pentasulfide as a novel additive for high-performance lithium-sulfur batteries

    SciTech Connect

    Lin, Zhan; Liu, Zengcai; Fu, Wujun; Dudney, Nancy J; Liang, Chengdu

    2012-01-01

    Lithium-sulfur (Li-S) batteries suffer from rapid capacity decay and low energy efficiency because of the low solubility of lithium sulfide (Li2S) in organic solvents and the intrinsic polysulfide shuttle phenomenon. Here, we report a novel additive of phosphorus pentasulfide (P2S5) in organic electrolyte to boost the cycling performance of Li-S batteries. The function of the additive is two-fold: (1) P2S5 promotes the dissolution of Li2S and alleviates the loss of capacity caused by the precipitation of Li2S; and (2) P2S5 passivates the surface of lithium metal and therefore eliminates the polysulfide shuttle phenomenon. A Li-S test cell demonstrates high reversible capacity of 900-1350 mAh g-1 and high coulombic efficiency of 90% for 40 stable cycles at 0.1 C.

  8. Biofiltration of high formaldehyde loads with ozone additions in long-term operation.

    PubMed

    Maldonado-Diaz, G; Arriaga, S

    2015-01-01

    Formaldehyde (FA) biofiltration was evaluated over 310 days with and without ozone addition. Without ozone, the biofilter was able to treat formaldehyde at inlet loads (ILs) lower than 40 g m(-3) h(-1), maintaining, under this condition, an average removal efficiency (RE) of 88 % for a few days before collapsing to zero. The continuous addition of ozone (90 ppbv) helped to recover the RE from zero to 98 ± 2 % and made it possible to operate at an IL of 40 g m(-3) h(-1) for long periods of operation (107 days). Furthermore, the ozone addition aided in operating the biofilter at a formaldehyde IL of up to 120 g m(-3) h(-1) values that have never before been reached. GC-mass spectrometry (MS) analysis showed that dimethoxymethane was the common compound in leachate during the performance decay. Also, the addition of ozone aided in maintaining an optimal pH in the biofilter with values between 7.5 and 8.2, due to the carbonate species formed during the ozone reactions with formaldehyde and its by-products. Thus, the pH control was confirmed and the alkalinity of the biofilter increased from 334.1 ± 100.3 to 1450 ± 127 mg CaCO3 L(-1) when ozone was added. Ozone addition diminished the exopolymeric substances (EPS) content of biofilm and biofilm thickness without affecting cell viability. Kinetic parameters suggested that the best conditions for carrying out FA biofiltration were reached under ozone addition. The addition of ozone during formaldehyde biofiltration could be a good strategy to maintain the pH and the steady state of the system under high ILs and for long periods of operation.

  9. The effect of additional high dose carbon implantation on the tribological properties of titanium implanted steel

    NASA Astrophysics Data System (ADS)

    Hayashi, Kazunori; Sasaki, Jun; Ichiko, Osami; Hashiguchi, Yoshihiro

    1996-08-01

    The tribological properties and the structural changes of hardened steel implanted with titanium followed by carbon were investigated as a function of additional carbon dose. The dose of Ti was 5×10 17 Ti cm -2 and the additional C doses were 0, 4×10 17, 8×10 17 and 1.2×10 18 C cm -2. After Ti implantation, the steel surface transformed to a FeTiC ternary amorphous phase. Additional implantation of carbon to a dose of 4×10 17 C cm -2 produced fine TiC precipitates dispersed in the ternary amorphous matrix. When the additional C dose exceeded 8×10 17 C cm -2, very fine graphite precipitates appeared in the ternary amorphous phase. The steel surface with very fine graphite precipitates exhibited superior tribological properties. The benefits provided by additional high dose carbon implantation are considered as follows: strengthening of the amorphous phase, thickening of the modified layer, dispersion strengthening of the implanted layer by very fine graphite precipitates and lubrication effect by graphite particles. Comparing the friction properties of Ti+C implanted steel with that of C implanted steel, the role of Ti implantation is to reduce the friction of the surface during sliding and the role of C implantation is to increase the lifetime of the surface against wear.

  10. Bioremediation of high organic load lagoon sediments: compost addition and priming effects.

    PubMed

    d'Errico, G; Giovannelli, D; Montano, C; Milanovic, V; Ciani, M; Manini, E

    2013-03-01

    Lagoons are often affected by eutrophication phenomena, due to their shallow nature, high productivity, weak hydrodynamism and anthropic exploitation. Bioremediation techniques have been widely used in the treatment of chemical pollution; however, no information is available on the use of bioremediation of organic-rich sediments. In the present study, we investigated the priming effects following compost addition to organic-rich lagoon sediments, and the effects of this compost addition on degradation and cycling of organic detritus, transfer of organic matter to higher trophic levels, and in situ prokaryotic community structure. There was a positive response to treatment, particularly during the first days after compost addition. The compost had a stimulating effect on degradation activity of the prokaryotic community. This occurred despite an increase in available organic matter, as the community was more efficient at removing it. These data are supported by the prokaryotic community structure analysis, which revealed no changes in the in situ community following compost addition. This priming effect enhancement through compost addition represents an efficient method to treat organic-rich sediments. PMID:23273326

  11. High Salt Diet Affects Renal Sodium Excretion and ERRα Expression

    PubMed Central

    Wang, Dan; Wang, Yang; Liu, Fu-Qiang; Yuan, Zu-Yi; Mu, Jian-Jun

    2016-01-01

    Kidneys regulate the balance of water and sodium and therefore are related to blood pressure. It is unclear whether estrogen-related receptor α (ERRα), an orphan nuclear receptor and transcription factor highly expressed in kidneys, affects the reabsorption of water and sodium. The aim of this study was to determine whether changes in the expressions of ERRα, Na+/K+-ATPase and epithelial sodium channel (ENaC) proteins affected the reabsorption of water and sodium in kidneys of Dahl salt-sensitive (DS) rats. SS.13BN rats, 98% homologous to the DS rats, were used as a normotensive control group. The 24 h urinary sodium excretion of the DS and SS.13BN rats increased after the 6-week high salt diet intervention, while sodium excretion was increased in DS rats with daidzein (agonist of ERRα) treatment. ERRα expression was decreased, while β- and γ-ENaC mRNA expressions were increased upon high sodium diet treatment in the DS rats. In the chromatin immunoprecipitation (CHIP) assay, positive PCR signals were obtained in samples treated with anti-ERRα antibody. The transcriptional activity of ERRα was decreased upon high salt diet intervention. ERRα reduced the expressions of β- and γ-ENaC by binding to the ENaC promoter, thereby increased Na+ reabsorption. Therefore, ERRα might be one of the factors causing salt-sensitive hypertension. PMID:27043552

  12. High Salt Diet Affects Renal Sodium Excretion and ERRα Expression.

    PubMed

    Wang, Dan; Wang, Yang; Liu, Fu-Qiang; Yuan, Zu-Yi; Mu, Jian-Jun

    2016-01-01

    Kidneys regulate the balance of water and sodium and therefore are related to blood pressure. It is unclear whether estrogen-related receptor α (ERRα), an orphan nuclear receptor and transcription factor highly expressed in kidneys, affects the reabsorption of water and sodium. The aim of this study was to determine whether changes in the expressions of ERRα, Na⁺/K⁺-ATPase and epithelial sodium channel (ENaC) proteins affected the reabsorption of water and sodium in kidneys of Dahl salt-sensitive (DS) rats. SS.13BN rats, 98% homologous to the DS rats, were used as a normotensive control group. The 24 h urinary sodium excretion of the DS and SS.13BN rats increased after the 6-week high salt diet intervention, while sodium excretion was increased in DS rats with daidzein (agonist of ERRα) treatment. ERRα expression was decreased, while β- and γ-ENaC mRNA expressions were increased upon high sodium diet treatment in the DS rats. In the chromatin immunoprecipitation (CHIP) assay, positive PCR signals were obtained in samples treated with anti-ERRα antibody. The transcriptional activity of ERRα was decreased upon high salt diet intervention. ERRα reduced the expressions of β- and γ-ENaC by binding to the ENaC promoter, thereby increased Na+ reabsorption. Therefore, ERRα might be one of the factors causing salt-sensitive hypertension. PMID:27043552

  13. Organocatalyzed cascade aza-Michael/Michael addition for the asymmetric construction of highly functionalized spiropyrazolone tetrahydroquinolines.

    PubMed

    Li, Jun-Hua; Du, Da-Ming

    2014-11-01

    An organocatalyzed diastereo- and enantioselective cascade aza-Michael/Michael addition of 2-tosylaminoenones to unsaturated pyrazolones has been developed to afford novel chiral spiropyrazolone tetrahydroquinolines containing three contiguous stereocenters. This cascade reaction proceeded well with 2 mol% chiral bifunctional tertiary amine squaramide catalyst to give the desired products in excellent yields (up to 99%) with excellent diastereoselectivity (up to >25:1 diastereomeric ratio) and high enantioselectivity (up to 91% enantiomeric excess).

  14. Hydrodynamic Instability in High-speed Direct Laser Deposition for Additive Manufacturing

    NASA Astrophysics Data System (ADS)

    Turichin, Gleb; Zemlyakov, Evgeny; Klimova, Olga; Babkin, Konstantin

    High speed direct laser deposition, when product forms from metal powder, transferred by gas-powder jet, supplied coaxially or non-coaxially to focused laser beam, in one of most prospective additive technologies for production parts for aircraft engines. The limit of process productivity is connected with development of hydrodynamic instability of the melt pool in conditions of high power laser action and material supply by gas-powder jet. Theoretical analysis and experiments allowed clarified a physical nature of instability appearance, determine a stability conditions and invent a methods which allow avoid instability in deposition process. Nozzles for direct laser deposition, designed with consideration of stability conditions, allow get a level of process productivity more then 2 kg/h. The developed technology of deposition and technological equipment, based on high power fiber laser, has been used for manufacturing of parts for "high temperature" unit of aircraft engine.

  15. Dopamine as a Novel Electrolyte Additive for High-Voltage Lithium-Ion Batteries.

    PubMed

    Lee, Hoogil; Han, Taeyeong; Cho, Kuk Young; Ryou, Myung-Hyun; Lee, Yong Min

    2016-08-24

    Dopamine, which can be electrochemically oxidized to polydopamine on cathode surface, was introduced as an electrolyte additive for high-voltage lithium-ion batteries (LIBs). The addition of 0.1 wt % dopamine to the electrolyte led to the formation of a polydopamine-containing layer on the cathode, thereby resulting in suppression of the oxidative decomposition of the electrolyte during high-voltage operation (up to 4.5 V) of a LiNi1/3Co1/3Mn1/3O2/artificial graphite cell. The addition of dopamine to the electrolyte improved the capacity retention of the cell from 136 to 147 mAh g(-1) after 100 cycles at a rate of 1 C and a cutoff voltage of 4.5 V, while the cycle performance and rate capability with a cutoff voltage of 4.3 V were comparable to those of the cell without dopamine. Further evidence of the positive impact of dopamine on high-voltage LIBs was the lower DC-IRs and AC impedances, as well as the retention of the cathode morphology even after operation at 4.5 V. PMID:27509406

  16. Dopamine as a Novel Electrolyte Additive for High-Voltage Lithium-Ion Batteries.

    PubMed

    Lee, Hoogil; Han, Taeyeong; Cho, Kuk Young; Ryou, Myung-Hyun; Lee, Yong Min

    2016-08-24

    Dopamine, which can be electrochemically oxidized to polydopamine on cathode surface, was introduced as an electrolyte additive for high-voltage lithium-ion batteries (LIBs). The addition of 0.1 wt % dopamine to the electrolyte led to the formation of a polydopamine-containing layer on the cathode, thereby resulting in suppression of the oxidative decomposition of the electrolyte during high-voltage operation (up to 4.5 V) of a LiNi1/3Co1/3Mn1/3O2/artificial graphite cell. The addition of dopamine to the electrolyte improved the capacity retention of the cell from 136 to 147 mAh g(-1) after 100 cycles at a rate of 1 C and a cutoff voltage of 4.5 V, while the cycle performance and rate capability with a cutoff voltage of 4.3 V were comparable to those of the cell without dopamine. Further evidence of the positive impact of dopamine on high-voltage LIBs was the lower DC-IRs and AC impedances, as well as the retention of the cathode morphology even after operation at 4.5 V.

  17. Improving sulfolane-based electrolyte for high voltage Li-ion cells with electrolyte additives

    NASA Astrophysics Data System (ADS)

    Xia, Jian; Dahn, J. R.

    2016-08-01

    An electrolyte mixture containing 1 M LiPF6 in sulfolane:ethylmethyl carbonate 3:7 with vinylene carbonate and other electrolyte additives exhibited promising cycling and storage performance in high voltage Li(Ni0·4Mn0·4Co0.2)O2/graphite pouch type Li-ion cells tested to 4.5 V. Voltage drop during storage, coulombic efficiency, charge endpoint capacity slippage during ultra high precision cycling, charge-transfer resistance after storage or cycling, gas evolution during storage and cycling as well as capacity retention during long-term cycling were examined. The results for cells with sulfolane-based electrolytes were compared with those for cells with ethylene carbonate-based electrolytes containing state-of-the-art electrolyte additives. This survey showed that the combination of vinylene carbonate and triallyl phosphate as electrolyte additives in sulfolane:ethylmethyl carbonate electrolyte yielded cells capable of better performance during tests to 4.5 V than cells with ethylene carbonate-based electrolytes. These results suggest that sulfolane-based electrolytes may be promising for high voltage Li-ion cells.

  18. Improving sulfolane-based electrolyte for high voltage Li-ion cells with electrolyte additives

    NASA Astrophysics Data System (ADS)

    Xia, Jian; Dahn, J. R.

    2016-08-01

    An electrolyte mixture containing 1 M LiPF6 in sulfolane:ethylmethyl carbonate 3:7 with vinylene carbonate and other electrolyte additives exhibited promising cycling and storage performance in high voltage Li(Ni0·4Mn0·4Co0.2)O2/graphite pouch type Li-ion cells tested to 4.5 V. Voltage drop during storage, coulombic efficiency, charge endpoint capacity slippage during ultra high precision cycling, charge-transfer resistance after storage or cycling, gas evolution during storage and cycling as well as capacity retention during long-term cycling were examined. The results for cells with sulfolane-based electrolytes were compared with those for cells with ethylene carbonate-based electrolytes containing state-of-the-art electrolyte additives. This survey showed that the combination of vinylene carbonate and triallyl phosphate as electrolyte additives in sulfolane:ethylmethyl carbonate electrolyte yielded cells capable of better performance during tests to 4.5 V than cells with ethylene carbonate-based electrolytes. These results suggest that sulfolane-based electrolytes may be promising for high voltage Li-ion cells.

  19. High expression of RUNX1 is associated with poorer outcomes in cytogenetically normal acute myeloid leukemia

    PubMed Central

    Xu, Keman; Hu, Kai; Wang, Jing; Wang, Jijun; Jing, Hongmei; Shi, Jinlong; Ke, Xiaoyan

    2016-01-01

    Depending on its expression level, RUNX1 can act as a tumor promoter or suppressor in hematological malignancies. The clinical impact of RUNX1 expression in cytogenetically normal acute myeloid leukemia (CN-AML) remained unknown, however. We evaluated the prognostic significance of RUNX1 expression using several public microarray datasets. In the testing group (n = 157), high RUNX1 expression (RUNX1high) was associated with poorer overall survival (OS; P = 0.0025) and event-free survival (EFS; P = 0.0025) than low RUNX1 expression (RUNX1low). In addition, the prognostic significance of RUNX1 was confirmed using European Leukemia Net (ELN) genetic categories and multivariable analysis, which was further validated using a second independent CN-AML cohort (n = 162, OS; P = 0.03953). To better understand the mechanisms of RUNX1, we investigated genome-wide gene/microRNAs expression signatures and cell signaling pathways associated with RUNX1 expression status. Several known oncogenes/oncogenic microRNAs and cell signaling pathways were all up-regulated, while some anti-oncogenes and molecules of immune activation were down-regulated in RUNX1high CN-AML patients. These findings suggest RUNX1high is a prognostic biomarker of unfavorable outcome in CN-AML, which is supported by the distinctive gene/microRNA signatures and cell signaling pathways. PMID:26910834

  20. Modulation of chicken intestinal immune gene expression by small cationic peptides as feed additives during the first week posthatch.

    PubMed

    Kogut, Michael H; Genovese, Kenneth J; He, Haiqi; Swaggerty, Christina L; Jiang, Yiwei

    2013-09-01

    We have been investigating modulation strategies tailored around the selective stimulation of the host's immune system as an alternative to direct targeting of microbial pathogens by antibiotics. One such approach is the use of a group of small cationic peptides (BT) produced by a Gram-positive soil bacterium, Brevibacillus texasporus. These peptides have immune modulatory properties that enhance both leukocyte functional efficiency and leukocyte proinflammatory cytokine and chemokine mRNA transcription activities in vitro. In addition, when provided as a feed additive for just 4 days posthatch, BT peptides significantly induce a concentration-dependent protection against cecal and extraintestinal colonization by Salmonella enterica serovar Enteritidis. In the present studies, we assessed the effects of feeding BT peptides on transcriptional changes on proinflammatory cytokines, inflammatory chemokines, and Toll-like receptors (TLR) in the ceca of broiler chickens with and without S. Enteritidis infection. After feeding a BT peptide-supplemented diet for the first 4 days posthatch, chickens were then challenged with S. Enteritidis, and intestinal gene expression was measured at 1 or 7 days postinfection (p.i.) (5 or 11 days of age). Intestinal expression of innate immune mRNA transcripts was analyzed by quantitative real-time PCR (qRT-PCR). Analysis of relative mRNA expression showed that a BT peptide-supplemented diet did not directly induce the transcription of proinflammatory cytokine, inflammatory chemokine, type I/II interferon (IFN), or TLR mRNA in chicken cecum. However, feeding the BT peptide-supplemented diet primed cecal tissue for increased (P ≤ 0.05) transcription of TLR4, TLR15, and TLR21 upon infection with S. Enteritidis on days 1 and 7 p.i. Likewise, feeding the BT peptides primed the cecal tissue for increased transcription of proinflammatory cytokines (interleukin 1β [IL-1β], IL-6, IL-18, type I and II IFNs) and inflammatory chemokine (CxCLi2

  1. Processing false solutions in additions: differences between high- and lower-skilled arithmetic problem-solvers.

    PubMed

    Núñez-Peña, Maria Isabel; Suárez-Pellicioni, Macarena

    2012-05-01

    This paper focuses on the capacity to solve numerical incongruities in high- and lower-skilled arithmetic problem-solvers by investigating event-related brain potentials elicited by incorrect solutions to additions. Fifteen high-skill and fifteen low-skill individuals were presented with simple addition problems in a verification task. The proposed solution was manipulated by presenting correct solutions and incorrect solutions very close to the correct ones. Incorrect solutions elicited a negative component followed by a late positive component (LPC/P3b), whose amplitude was smaller for the low-skill group than for the high-skill group. Because the LPC/P3b amplitude has been taken as an indicator of the plausibility of the stimulus, this result suggests that incorrect solutions close to the correct ones appear more plausible to low-skilled individuals than to their high-skilled counterparts. This result is interpreted in terms of differences in the strength of association between problems and potential solutions depending on arithmetical skill.

  2. Additive transgene expression and genetic introgression in multiple green-fluorescent protein transgenic crop x weed hybrid generations.

    PubMed

    Halfhill, M D; Millwood, R J; Weissinger, A K; Warwick, S I; Stewart, C N

    2003-11-01

    The level of transgene expression in crop x weed hybrids and the degree to which crop-specific genes are integrated into hybrid populations are important factors in assessing the potential ecological and agricultural risks of gene flow associated with genetic engineering. The average transgene zygosity and genetic structure of transgenic hybrid populations change with the progression of generations, and the green fluorescent protein (GFP) transgene is an ideal marker to quantify transgene expression in advancing populations. The homozygous T(1) single-locus insert GFP/ Bacillus thuringiensis (Bt) transgenic canola ( Brassica napus, cv Westar) with two copies of the transgene fluoresced twice as much as hemizygous individuals with only one copy of the transgene. These data indicate that the expression of the GFP gene was additive, and fluorescence could be used to determine zygosity status. Several hybrid generations (BC(1)F(1), BC(2)F(1)) were produced by backcrossing various GFP/Bt transgenic canola ( B. napus, cv Westar) and birdseed rape ( Brassica rapa) hybrid generations onto B. rapa. Intercrossed generations (BC(2)F(2) Bulk) were generated by crossing BC(2)F(1) individuals in the presence of a pollinating insect ( Musca domestica L.). The ploidy of plants in the BC(2)F(2) Bulk hybrid generation was identical to the weedy parental species, B. rapa. AFLP analysis was used to quantify the degree of B. napus introgression into multiple backcross hybrid generations with B. rapa. The F(1) hybrid generations contained 95-97% of the B. napus-specific AFLP markers, and each successive backcross generation demonstrated a reduction of markers resulting in the 15-29% presence in the BC(2)F(2) Bulk population. Average fluorescence of each successive hybrid generation was analyzed, and homozygous canola lines and hybrid populations that contained individuals homozygous for GFP (BC(2)F(2) Bulk) demonstrated significantly higher fluorescence than hemizygous hybrid

  3. Additive transgene expression and genetic introgression in multiple green-fluorescent protein transgenic crop x weed hybrid generations.

    PubMed

    Halfhill, M D; Millwood, R J; Weissinger, A K; Warwick, S I; Stewart, C N

    2003-11-01

    The level of transgene expression in crop x weed hybrids and the degree to which crop-specific genes are integrated into hybrid populations are important factors in assessing the potential ecological and agricultural risks of gene flow associated with genetic engineering. The average transgene zygosity and genetic structure of transgenic hybrid populations change with the progression of generations, and the green fluorescent protein (GFP) transgene is an ideal marker to quantify transgene expression in advancing populations. The homozygous T(1) single-locus insert GFP/ Bacillus thuringiensis (Bt) transgenic canola ( Brassica napus, cv Westar) with two copies of the transgene fluoresced twice as much as hemizygous individuals with only one copy of the transgene. These data indicate that the expression of the GFP gene was additive, and fluorescence could be used to determine zygosity status. Several hybrid generations (BC(1)F(1), BC(2)F(1)) were produced by backcrossing various GFP/Bt transgenic canola ( B. napus, cv Westar) and birdseed rape ( Brassica rapa) hybrid generations onto B. rapa. Intercrossed generations (BC(2)F(2) Bulk) were generated by crossing BC(2)F(1) individuals in the presence of a pollinating insect ( Musca domestica L.). The ploidy of plants in the BC(2)F(2) Bulk hybrid generation was identical to the weedy parental species, B. rapa. AFLP analysis was used to quantify the degree of B. napus introgression into multiple backcross hybrid generations with B. rapa. The F(1) hybrid generations contained 95-97% of the B. napus-specific AFLP markers, and each successive backcross generation demonstrated a reduction of markers resulting in the 15-29% presence in the BC(2)F(2) Bulk population. Average fluorescence of each successive hybrid generation was analyzed, and homozygous canola lines and hybrid populations that contained individuals homozygous for GFP (BC(2)F(2) Bulk) demonstrated significantly higher fluorescence than hemizygous hybrid

  4. [Influence Additional Cognitive Tasks on EEG Beta Rhythm Parameters during Forming and Testing Set to Perception of the Facial Expression].

    PubMed

    Yakovenko, I A; Cheremushkin, E A; Kozlov, M K

    2015-01-01

    The research of changes of a beta rhythm parameters on condition of working memory loading by extension of a interstimuli interval between the target and triggering stimuli to 16 sec is investigated on 70 healthy adults in two series of experiments with set to a facial expression. In the second series at the middle of this interval for strengthening of the load was entered the additional cognitive task in the form of conditioning stimuli like Go/NoGo--circles of blue or green color. Data analysis of the research was carried out by means of continuous wavelet-transformation on the basis of "mather" complex Morlet-wavelet in the range of 1-35 Hz. Beta rhythm power was characterized by the mean level, maxima of wavelet-transformation coefficient (WLC) and latent periods of maxima. Introduction of additional cognitive task to pause between the target and triggering stimuli led to essential increase in absolute values of the mean level of beta rhythm WLC and relative sizes of maxima of beta rhythm WLC. In the series of experiments without conditioning stimulus subjects with large number of mistakes (from 6 to 40), i.e. rigid set, in comparison with subjects with small number of mistakes (to 5), i.e. plastic set, at the forming stage were characterized by higher values of the mean level of beta rhythm WLC. Introduction of the conditioning stimuli led to smoothing of intergroup distinctions throughout the experiment. PMID:26601500

  5. Use of pyrocatechol violet as an effective redox additive for highly promoting the supercapacitor performances

    NASA Astrophysics Data System (ADS)

    Wang, Qian; Nie, Yong Fu; Chen, Xiang Ying; Xiao, Zheng Hui; Zhang, Zhong Jie

    2016-08-01

    In present work, we demonstrate a simple but effective redox additive of pyrocatechol violet (abbr. PCV) to largely promote the capacitive performances especially when carried out in three different kinds of electrolytes (H2SO4, Na2SO4 and KOH), mostly due to its fast electron and proton transfer occurring in the electrode/electrolyte interface. It reveals that the PCV dosage incorporated into electrolyte plays a crucial role in the determination of capacitive performance. When conducted in a two-electrode system, incorporating 0.06 mol L-1 PCV into 1 mol L-1 H2SO4 can achieve large capacitance up to 200 F g-1, which is almost 2.06 times than the one without the addition of PCV; besides, the corresponding energy density is of 28 Wh kg-1 (also increasing 2 times). What's more, PCV has been extended as the redox additive in other electrolytes such as Na2SO4 and KOH, and remarkable promotion in capacitance and energy density also occur, well evincing the high efficiency and universal applicability of PCV for the large promotion of supercapacitors' performances due to its prominent electrochemical reversibility and high solubility.

  6. High expression of NPY receptors in the human testis.

    PubMed

    Körner, Meike; Waser, Beatriche; Thalmann, George N; Reubii, Jean Claude

    2011-04-30

    NPY receptors represent novel molecular therapeutic targets in cancer and obesity. However, the extent of NPY receptor expression in normal human tissues is poorly investigated. Based on the role of NPY in reproductive functions, the NPY receptor expression was studied in 25 normal human testes and, additionally, 24 testicular tumors using NPY receptor autoradiography. In the normal testis, Leydig cells strongly expressed NPY receptor subtype Y2, and small arterial blood vessels Y1. Y2 receptors were found to be functional with agonist-stimulated [(35)S]GTPγS binding autoradiography. Full functional integrity of the NPY system was further suggested by the immunohistochemical detection of NPY peptide in nerve fibers directly adjacent to Leydig cells and arteries. Germ cell tumors expressed Y1 and Y2 on tumor cells in 33% and Y1 on intratumoral blood vessels in 50%. Based on its strong NPY receptor expression in Leydig cells and blood vessels, the normal human testis represents a potentially important physiological and pharmalogical NPY target.

  7. Low TGFβ1 expression prevents and high expression exacerbates diabetic nephropathy in mice

    PubMed Central

    Hathaway, Catherine K.; Gasim, Adil M. H.; Grant, Ruriko; Chang, Albert S.; Kim, Hyung-Suk; Madden, Victoria J.; Bagnell, C. Robert; Jennette, J. Charles; Smithies, Oliver; Kakoki, Masao

    2015-01-01

    Nephropathy develops in many but not all patients with long-standing type 1 diabetes. Substantial efforts to identify genotypic differences explaining this differential susceptibility have been made, with limited success. Here, we show that the expression of the transforming growth factor β1 gene (Tgfb1) affects the development of diabetic nephropathy in mice. To do this we genetically varied Tgfb1 expression in five steps, 10%, 60%, 100%, 150%, and 300% of normal, in mice with type 1 diabetes caused by the Akita mutation in the insulin gene (Ins2Akita). Although plasma glucose levels were not affected by Tgfb1 genotype, many features of diabetic nephropathy (mesangial expansion, elevated plasma creatinine and urea, decreased creatinine clearance and albuminuria) were progressively ameliorated as Tgfb1 expression decreased and were progressively exacerbated when expression was increased. The diabetic 10% hypomorphs had comparable creatinine clearance and albumin excretion to wild-type mice and no harmful changes in renal morphology. The diabetic 300% hypermorphs had ∼1/3 the creatinine clearance of wild-type mice, >20× their albumin excretion, ∼3× thicker glomerular basement membranes and severe podocyte effacement, matching human diabetic nephropathy. Switching Tgfb1 expression from low to high in the tubules of the hypomorphs increased their albumin excretion more than 10-fold but creatinine clearance remained high. Switching Tgfb1 expression from low to high in the podocytes markedly decreased creatinine clearance, but minimally increased albumin excretion. Decreasing expression of Tgfb1 could be a promising option for preventing loss of renal function in diabetes. PMID:25902541

  8. Communication: Virial coefficients and demixing in highly asymmetric binary additive hard-sphere mixtures.

    PubMed

    López de Haro, Mariano; Tejero, Carlos F; Santos, Andrés

    2013-04-28

    The problem of demixing in a binary fluid mixture of highly asymmetric additive hard spheres is revisited. A comparison is presented between the results derived previously using truncated virial expansions for three finite size ratios with those that one obtains with the same approach in the extreme case in which one of the components consists of point particles. Since this latter system is known not to exhibit fluid-fluid segregation, the similarity observed for the behavior of the critical constants arising in the truncated series in all instances, while not being conclusive, may cast serious doubts as to the actual existence of a demixing fluid-fluid transition in disparate-sized binary additive hard-sphere mixtures.

  9. Winter-time CO2 addition in high rate algal mesocosms for enhanced microalgal performance.

    PubMed

    Sutherland, Donna L; Montemezzani, Valerio; Mehrabadi, Abbas; Craggs, Rupert J

    2016-02-01

    Carbon limitation in domestic wastewater high rate algal ponds is thought to constrain microalgal photo-physiology and productivity and CO2 augmentation is often used to overcome this limitation in summer. However, the implications of carbon limitation during winter are poorly understood. This paper investigates the effects of 0.5%, 2%, 5% and 10% CO2 addition on the winter-time performance of wastewater microalgae in high rate algal mesocosms. Performance was measured in terms of light absorption, photosynthetic efficiency, biomass production and nutrient removal rates, along with community composition. Varying percentage CO2 addition and associated change in culture pH resulted in 3 distinct microalgal communities. Light absorption by the microalgae increased by up to 144% with CO2 addition, while a reduction in the package effect meant that there was less internal self-shading thereby increasing the efficiency of light absorption. Carbon augmentation increased the maximum rate of photosynthesis by up to 172%, which led to increased microalgal biovolume by up to 181% and an increase in total organic biomass for all treatments except 10% CO2. While 10% CO2 improved light absorption and photosynthesis this did not translate to enhanced microalgal productivity. Increased microalgal productivity with CO2 addition did not result in increased dissolved nutrient (nitrogen and phosphorus) removal. This experiment demonstrated that winter-time carbon augmentation up to 5% CO2 improved microalgal light absorption and utilisation, which ultimately increased microalgal biomass and is likely to enhance total annual microalgal areal productivity in HRAPs.

  10. Polymorphism in paracetamol: evidence of additional forms IV and V at high pressure.

    PubMed

    Smith, Spencer J; Bishop, Matthew M; Montgomery, Jeffrey M; Hamilton, Tracy P; Vohra, Yogesh K

    2014-08-01

    The structural phase stability of N-(4-hydroxyphenyl) acetamide (paracetamol) has been studied at ambient temperature up to 23 GPa using Raman spectroscopy. Spectral changes have provided further evidence for a highly kinetically driven Form I → II transition that occurs as a mixed phase from 4.8 to 6.5 GPa, and might complete as early as 7 GPa. Upon further compression to 8.1 GPa, a drastic shift in spectral signature was observed providing the first evidence for a previously undiscovered Form IV of paracetamol. Additional shifts in mode intensities were observed near 11 GPa indicating a potential restructuring of the hydrogen bonding network and/or structural modification to a potentially new Form V. Phase boundaries at 7 and 8 GPa were confirmed under hydrostatic conditions using Raman spectroscopy. Spectral changes indicate that the transition Form IV → V occurs near 11 GPa. Multiple ab initio harmonic frequency calculations at different levels of theory were performed with a B3LYP/6-31G** being used to provide a more robust mode assignment to our experimentally obtained Raman modes. High pressure X-ray diffraction (XRD) was performed up to 21 GPa, which provided further evidence for a highly kinetically driven Form I → II transition in agreement with our Raman measurements. In addition, the XRD provided further evidence for the existence of Form IV near 8 GPa and Form V near 11 GPa with Form V persisting up to 21 GPa.

  11. Calcium Carbonate Nanoplate Assemblies with Directed High-Energy Facets: Additive-Free Synthesis, High Drug Loading, and Sustainable Releasing.

    PubMed

    Zhang, Jing; Li, Yu; Xie, Hao; Su, Bao-Lian; Yao, Bin; Yin, Yixia; Li, Shipu; Chen, Fang; Fu, Zhengyi

    2015-07-29

    Developing drug delivery systems (DDSs) with high drug-loading capacity and sustainable releasing is critical for long-term chemotherapeutic efficacy, and it still remains challenging. Herein, vaterite CaCO3 nanoplate assemblies with exposed high-energy {001} facets have been synthesized via a novel, additive-free strategy. The product shows a high doxorubicin-loading capacity (65%); the best of all the CaCO3-based DDSs so far. Also, the product's sustainable releasing performance and its inhibition of the initial burst release, together, endow it with long-term drug efficacy. The work may shed light on exposing directed high-energy facets for rationally designing of a drug delivery system with long-term efficacy.

  12. Investigations of non-linear polymers as high performance lubricant additives

    SciTech Connect

    Robinson, Joshua W.; Bhattacharya, Priyanka; Qu, Jun; Bays, J. Timothy; Cosimbescu, Lelia

    2015-03-22

    Off-the-shelf available engine oils contain an assortment of additives that increase the performance of base oils and maximize the overall efficiency of the machine. With ever increasing requirements for fuel efficiency, the demand for novel materials that outperform older generations is also on the rise. One approach towards increasing overall efficiency is to reduce internal friction and wear in an engine. From an additive approach, this is typically achieved by altering the bulk oil’s viscosity at high temperatures via polymers. In general, the hydrodynamic volume of polymers increase (expand) at elevated temperatures and decrease (contract/deflate) with declining temperatures and this effect is enhanced be carefully designing specific structures and architectures. The natural thinning tendency of base oil with increasing temperatures is in part mitigated by the expansion of the macromolecules added, and the overall effect is decreasing the viscosity losses at high temperatures. Traditional polymer architectures vary from linear to dendritic, where linear polymers of the same chemical composition and molecular weight to its dendritic counterpart will undergo a more significant free volume change in solution with regards to temperature changes. This advantage has been exploited in the literature towards the production of viscosity modifiers. However, one major disadvantage of linear polymers is degradation due to mechanical shear forces and high temperatures causing a shorter additive lifetime. Dendrimers on the other hand are known to demonstrate superior robustness to shear degradation when compared to their respective linear counterparts. An additional advantage of the dendritic architecture is the ability to tailor the peripheral end-groups towards influencing polymer-solvent and/or polymer-surface interactions. Comb-burst hyperbranched polymers are a hybrid of the aforementioned architectures and provide several compromises between the traditional

  13. [INVITED] Laser-induced forward transfer: A high resolution additive manufacturing technology

    NASA Astrophysics Data System (ADS)

    Delaporte, Philippe; Alloncle, Anne-Patricia

    2016-04-01

    Among the additive manufacturing techniques, laser-induced forward transfer addresses the challenges of printing thin films in solid phase or small volume droplets in liquid phase with very high resolution. This paper reviews the physics of this process and explores the pros and cons of this technology versus other digital printing technologies. The main field of applications are printed electronics, organic electronics and tissue engineering, and the most promising short terms ones concern digital laser printing of sensors and conductive tracks. Future directions and emerging areas of interest are discussed such as printing solid from a liquid phase and 3D digital nanomanufacturing.

  14. Determination of plastic additives in packaging by liquid chromatography coupled to high resolution mass spectrometry.

    PubMed

    Moreta, Cristina; Tena, María-Teresa

    2015-10-01

    A simple and sensitive analytical method for the determination of several plastic additives in multilayer packaging based on solid-liquid extraction (SLE) and ultra-high performance liquid chromatography (UHPLC) coupled to variable wavelength (VWD) and time of flight mass spectrometry (TOF-MS) detectors is presented. The proposed method allows the simultaneous determination of fourteen additives belonging to different families such as antioxidants, slip agents and light stabilizers, as well as two oxidation products in only 9min. The developed method was validated in terms of linearity, matrix effect error, detection and quantification limits, repeatability and intermediate precision. The instrumental method showed satisfactory repeatability and intermediate precision at concentrations closed to LOQ with RSDs less than 7 and 20%, respectively, and LODs until 5000 times more sensitive than other GC-FID and HPLC-VWD methods previously reported. Also, focused ultrasound solid-liquid extraction (FUSLE) was optimized and evaluated to extract plastic additives from packaging. Extraction results obtained by FUSLE and SLE were compared to those obtained by pressurized liquid extraction (PLE). All extraction methods showed excellent extraction efficiency for slip agents, however quantitative recovery of all analytes was achieved only by SLE with just 5ml of hexane for 10h. Finally, the selected method was applied to the analysis of packaging samples where erucamide, Irgafos 168, oxidized Irgafos 168, Irganox 1076 and Irganox 1010 were detected and quantified. PMID:26319625

  15. Determination of plastic additives in packaging by liquid chromatography coupled to high resolution mass spectrometry.

    PubMed

    Moreta, Cristina; Tena, María-Teresa

    2015-10-01

    A simple and sensitive analytical method for the determination of several plastic additives in multilayer packaging based on solid-liquid extraction (SLE) and ultra-high performance liquid chromatography (UHPLC) coupled to variable wavelength (VWD) and time of flight mass spectrometry (TOF-MS) detectors is presented. The proposed method allows the simultaneous determination of fourteen additives belonging to different families such as antioxidants, slip agents and light stabilizers, as well as two oxidation products in only 9min. The developed method was validated in terms of linearity, matrix effect error, detection and quantification limits, repeatability and intermediate precision. The instrumental method showed satisfactory repeatability and intermediate precision at concentrations closed to LOQ with RSDs less than 7 and 20%, respectively, and LODs until 5000 times more sensitive than other GC-FID and HPLC-VWD methods previously reported. Also, focused ultrasound solid-liquid extraction (FUSLE) was optimized and evaluated to extract plastic additives from packaging. Extraction results obtained by FUSLE and SLE were compared to those obtained by pressurized liquid extraction (PLE). All extraction methods showed excellent extraction efficiency for slip agents, however quantitative recovery of all analytes was achieved only by SLE with just 5ml of hexane for 10h. Finally, the selected method was applied to the analysis of packaging samples where erucamide, Irgafos 168, oxidized Irgafos 168, Irganox 1076 and Irganox 1010 were detected and quantified.

  16. Nutrient removal in wastewater treatment high rate algal ponds with carbon dioxide addition.

    PubMed

    Park, J B K; Craggs, R J

    2011-01-01

    The influence of CO2 addition to high rate algal ponds (HRAPS) on nitrogen removal was investigated using two pilot-scale HRAPs operated with different hydraulic retention times (HRT: 4 and 8 days), and was compared to the nitrogen removal by the 8-day HRT pond before CO2 addition was installed. Nitrogen balances were calculated by partitioning total nitrogen into organic and inorganic nitrogen (NH4+-N and NO3--N), and by separation of the organic nitrogen into particulate (PON) and dissolved organic nitrogen (DON). PON was further divided into algal organic nitrogen (AON) and bacteria organic nitrogen (BON) to investigate nitrogen mass flow in the HRAPS. This research shows that the proportion of algae in the algal/bacterial biomass in the longer 8-day HRT HRAP8d (55.6%) was appreciably lower than that in the shorter 4-day HRT HRAP4d (80.5%) when CO2 was added to control the maximum pH to <8.0 during the summer. Higher bacterial biomass in the longer 8-day HRT HRAP corresponded with higher nitrification rates, indicating that the longer 8-day HRT in the summer was detrimental for two reasons: lower algal productivity and increased nitrogen loss through nitrification/denitrification. Overall nitrogen removal of approximately 60% in the HRAPS with CO2 addition was mainly achieved by algal assimilation followed by sedimentation in the settling unit.

  17. A high-throughput microRNA expression profiling system.

    PubMed

    Guo, Yanwen; Mastriano, Stephen; Lu, Jun

    2014-01-01

    As small noncoding RNAs, microRNAs (miRNAs) regulate diverse biological functions, including physiological and pathological processes. The expression and deregulation of miRNA levels contain rich information with diagnostic and prognostic relevance and can reflect pharmacological responses. The increasing interest in miRNA-related research demands global miRNA expression profiling on large numbers of samples. We describe here a robust protocol that supports high-throughput sample labeling and detection on hundreds of samples simultaneously. This method employs 96-well-based miRNA capturing from total RNA samples and on-site biochemical reactions, coupled with bead-based detection in 96-well format for hundreds of miRNAs per sample. With low-cost, high-throughput, high detection specificity, and flexibility to profile both small and large numbers of samples, this protocol can be adapted in a wide range of laboratory settings. PMID:25030917

  18. Additive Manufacturing of Reactive In Situ Zr Based Ultra-High Temperature Ceramic Composites

    NASA Astrophysics Data System (ADS)

    Sahasrabudhe, Himanshu; Bandyopadhyay, Amit

    2016-03-01

    Reactive in situ multi-material additive manufacturing of ZrB2-based ultra-high-temperature ceramics in a Zr metal matrix was demonstrated using LENS™. Sound metallurgical bonding was achieved between the Zr metal and Zr-BN composites with Ti6Al4V substrate. Though the feedstock Zr power had α phase, LENS™ processing of the Zr powder and Zr-BN premix powder mixture led to the formation of some β phase of Zr. Microstructure of the Zr-BN composite showed primary grains of zirconium diboride phase in zirconium metal matrix. The presence of ZrB2 ceramic phase was confirmed by X-ray diffraction (XRD) analysis. Hardness of pure Zr was measured as 280 ± 12 HV and, by increasing the BN content in the feedstock, the hardness was found to increase. In Zr-5%BN composite, the hardness was 421 ± 10 HV and the same for Zr-10%BN composite was 562 ± 10 HV. It is envisioned that such multi-materials additive manufacturing will enable products in the future that cannot be manufactured using traditional approaches particularly in the areas of high-temperature metal-ceramic composites with compositional and functional gradation.

  19. Additive Manufacturing of Multifunctional Components Using High Density Carbon Nanotube Yarn Filaments

    NASA Technical Reports Server (NTRS)

    Gardner, John M.; Sauti, Godfrey; Kim, Jae-Woo; Cano, Roberto J.; Wincheski, Russell A.; Stelter, Christopher J.; Grimsley, Brian W.; Working, Dennis C.; Siochi, Emilie J.

    2016-01-01

    Additive manufacturing allows for design freedom and part complexity not currently attainable using traditional manufacturing technologies. Fused Filament Fabrication (FFF), for example, can yield novel component geometries and functionalities because the method provides a high level of control over material placement and processing conditions. This is achievable by extrusion of a preprocessed filament feedstock material along a predetermined path. However if fabrication of a multifunctional part relies only on conventional filament materials, it will require a different material for each unique functionality printed into the part. Carbon nanotubes (CNTs) are an attractive material for many applications due to their high specific strength as well as good electrical and thermal conductivity. The presence of this set of properties in a single material presents an opportunity to use one material to achieve multifunctionality in an additively manufactured part. This paper describes a recently developed method for processing continuous CNT yarn filaments into three-dimensional articles, and summarizes the mechanical, electrical, and sensing performance of the components fabricated in this way.

  20. [Simultaneous determination of six food additives in meat products by high performance liquid chromatography].

    PubMed

    Li, Xiuqin; Zhang, Qinghe; Yang, Zong

    2010-12-01

    A novel method was proposed for the simultaneous separation and determination of six food additives, benzoic acid, sorbic acid, saccharin sodium, acesulfame potassium, ponceau 4R and allura red AC, by high performance liquid chromatography (HPLC). After optimized the separation conditions of HPLC, the separation can be completed within 18 min by using a ZORBAX Eclipse Plus C18 column (150 mm x 4.6 mm, 5 microm) with 20 mmol/L ammonium acetate (pH 6.9) and methanol as the mobile phases. The gradient elution was performed by 8% methanol (0-2 min), 8%-50% methanol (2-3 min), 50% methanol (3-9 min), 50%-8% methanol (9-12 min) and 8% methanol (12-18 min). The detection wavelength was set at 235 nm. This method has been successfully applied to the analysis of meat products and the average recoveries ranged from 80.7% to 94.4% at high and low spiked levels. The relative standard deviations (RSDs, n=3) were between 2.0% and 7.1%. The method is simple, rapid, accurate and suitable for the simultaneous determination of the six food additives in meat products.

  1. Elite sport is not an additional source of distress for adolescents with high stress levels.

    PubMed

    Gerber, Markus; Holsboer-Trachsler, Edith; Pühse, Uwe; Brand, Serge

    2011-04-01

    This study examined whether participation in elite sport interacts with stress in decreasing or increasing symptoms of depression and anxiety among adolescents, and further, whether the interplay between participation in high-performance sport and stress is related to the perceived quality of sleep. 434 adolescents (278 girls, 156 boys; age: M = 17.2 yr.) from 15 "Swiss Olympic Sport Classes" and 9 conventional classes answered a questionnaire and completed a 7-day sleep log. Analyses of covariance showed that heightened stress was related to more depressive symptoms and higher scores for trait-anxiety. Moreover, those classified as having poor sleep by a median split cutoff reported higher levels of depressive symptoms. No significant (multivariate) main effects were found for high-performance sport athletes. Similarly, no significant two- or three-way interaction effects were found. These results caution against exaggerated expectations concerning sport participation as a stress buffer. Nevertheless, participation in high-performance sport was not found to be an additional source of distress for adolescents who reported high stress levels despite prior research that has pointed toward such a relationship.

  2. Multigroup Equivalence Analysis for High-Dimensional Expression Data

    PubMed Central

    Yang, Celeste; Bartolucci, Alfred A.; Cui, Xiangqin

    2015-01-01

    Hypothesis tests of equivalence are typically known for their application in bioequivalence studies and acceptance sampling. Their application to gene expression data, in particular high-dimensional gene expression data, has only recently been studied. In this paper, we examine how two multigroup equivalence tests, the F-test and the range test, perform when applied to microarray expression data. We adapted these tests to a well-known equivalence criterion, the difference ratio. Our simulation results showed that both tests can achieve moderate power while controlling the type I error at nominal level for typical expression microarray studies with the benefit of easy-to-interpret equivalence limits. For the range of parameters simulated in this paper, the F-test is more powerful than the range test. However, for comparing three groups, their powers are similar. Finally, the two multigroup tests were applied to a prostate cancer microarray dataset to identify genes whose expression follows a prespecified trajectory across five prostate cancer stages. PMID:26628859

  3. Arginine and proline applied as food additives stimulate high freeze tolerance in larvae of Drosophila melanogaster.

    PubMed

    Koštál, Vladimír; Korbelová, Jaroslava; Poupardin, Rodolphe; Moos, Martin; Šimek, Petr

    2016-08-01

    The fruit fly Drosophila melanogaster is an insect of tropical origin. Its larval stage is evolutionarily adapted for rapid growth and development under warm conditions and shows high sensitivity to cold. In this study, we further developed an optimal acclimation and freezing protocol that significantly improves larval freeze tolerance (an ability to survive at -5°C when most of the freezable fraction of water is converted to ice). Using the optimal protocol, freeze survival to adult stage increased from 0.7% to 12.6% in the larvae fed standard diet (agar, sugar, yeast, cornmeal). Next, we fed the larvae diets augmented with 31 different amino compounds, administered in different concentrations, and observed their effects on larval metabolomic composition, viability, rate of development and freeze tolerance. While some diet additives were toxic, others showed positive effects on freeze tolerance. Statistical correlation revealed tight association between high freeze tolerance and high levels of amino compounds involved in arginine and proline metabolism. Proline- and arginine-augmented diets showed the highest potential, improving freeze survival to 42.1% and 50.6%, respectively. Two plausible mechanisms by which high concentrations of proline and arginine might stimulate high freeze tolerance are discussed: (i) proline, probably in combination with trehalose, could reduce partial unfolding of proteins and prevent membrane fusions in the larvae exposed to thermal stress (prior to freezing) or during freeze dehydration; (ii) both arginine and proline are exceptional among amino compounds in their ability to form supramolecular aggregates which probably bind partially unfolded proteins and inhibit their aggregation under increasing freeze dehydration.

  4. Arginine and proline applied as food additives stimulate high freeze tolerance in larvae of Drosophila melanogaster.

    PubMed

    Koštál, Vladimír; Korbelová, Jaroslava; Poupardin, Rodolphe; Moos, Martin; Šimek, Petr

    2016-08-01

    The fruit fly Drosophila melanogaster is an insect of tropical origin. Its larval stage is evolutionarily adapted for rapid growth and development under warm conditions and shows high sensitivity to cold. In this study, we further developed an optimal acclimation and freezing protocol that significantly improves larval freeze tolerance (an ability to survive at -5°C when most of the freezable fraction of water is converted to ice). Using the optimal protocol, freeze survival to adult stage increased from 0.7% to 12.6% in the larvae fed standard diet (agar, sugar, yeast, cornmeal). Next, we fed the larvae diets augmented with 31 different amino compounds, administered in different concentrations, and observed their effects on larval metabolomic composition, viability, rate of development and freeze tolerance. While some diet additives were toxic, others showed positive effects on freeze tolerance. Statistical correlation revealed tight association between high freeze tolerance and high levels of amino compounds involved in arginine and proline metabolism. Proline- and arginine-augmented diets showed the highest potential, improving freeze survival to 42.1% and 50.6%, respectively. Two plausible mechanisms by which high concentrations of proline and arginine might stimulate high freeze tolerance are discussed: (i) proline, probably in combination with trehalose, could reduce partial unfolding of proteins and prevent membrane fusions in the larvae exposed to thermal stress (prior to freezing) or during freeze dehydration; (ii) both arginine and proline are exceptional among amino compounds in their ability to form supramolecular aggregates which probably bind partially unfolded proteins and inhibit their aggregation under increasing freeze dehydration. PMID:27489218

  5. Additive manufacturing of collagen scaffolds by three-dimensional plotting of highly viscous dispersions.

    PubMed

    Lode, Anja; Meyer, Michael; Brüggemeier, Sophie; Paul, Birgit; Baltzer, Hagen; Schröpfer, Michaela; Winkelmann, Claudia; Sonntag, Frank; Gelinsky, Michael

    2016-03-01

    Additive manufacturing (AM) allows the free form fabrication of three-dimensional (3D) structures with distinct external geometry, fitting into a patient-specific defect, and defined internal pore architecture. However, fabrication of predesigned collagen scaffolds using AM-based technologies is challenging due to the low viscosity of collagen solutions, gels or dispersions commonly used for scaffold preparation. In the present study, we have developed a straightforward method which is based on 3D plotting of a highly viscous, high density collagen dispersion. The swollen state of the collagen fibrils at pH 4 enabled the homogenous extrusion of the material, the deposition of uniform strands and finally the construction of 3D scaffolds. Stabilization of the plotted structures was achieved by freeze-drying and chemical crosslinking with the carbodiimide EDC. The scaffolds exhibited high shape and dimensional fidelity and a hierarchical porosity consisting of macropores generated by strand deposition as well as an interconnected microporosity within the strands as result of the freeze-drying process. Cultivation of human mesenchymal stromal cells on the scaffolds, with and without adipogenic or osteogenic stimulation, revealed their cytocompatibility and potential applicability for adipose and bone tissue engineering.

  6. Study of triallyl phosphate as an electrolyte additive for high voltage lithium-ion cells

    NASA Astrophysics Data System (ADS)

    Xia, J.; Madec, L.; Ma, L.; Ellis, L. D.; Qiu, W.; Nelson, K. J.; Lu, Z.; Dahn, J. R.

    2015-11-01

    The role of triallyl phosphate as an electrolyte additive in Li(Ni0.42Mn0.42Co0.16)O2/graphite pouch cells was studied using ex-situ gas measurements, ultra high precision coulometry, automated storage experiments, electrochemical impedance spectroscopy, long-term cycling and X-ray photoelectron spectroscopy. Cells containing triallyl phosphate produced less gas during formation, cycling and storage than control cells. The use of triallyl phosphate led to higher coulombic efficiency and smaller charge endpoint capacity slippage during ultra high precision charger testing. Cells containing triallyl phosphate showed smaller potential drop during 500 h storage at 40 °C and 60 °C and the voltage drop decreased as the triallyl phosphate content in the electrolyte increased. However, large amounts of triallyl phosphate (>3% by weight in the electrolyte) led to large impedance after cycling and storage. Symmetric cell studies showed large amounts of triallyl phosphate (5% or more) led to significant impedance increase at both negative and positive electrodes. X-ray photoelectron spectroscopy studies suggested that the high impedance came from the polymerization of triallyl phosphate molecules which formed thick solid electrolyte interphase films at the surfaces of both negative and positive electrodes. An optimal amount of 2%-3% triallyl phosphate led to better capacity retention during long term cycling.

  7. STAT4 Associates with SLE Through Two Independent Effects that Correlate with Gene Expression and Act Additively with IRF5 to Increase Risk

    PubMed Central

    Abelson, Anna-Karin; Delgado-Vega, Angélica M.; Kozyrev, Sergey V.; Sánchez, Elena; Velázquez-Cruz, Rafael; Eriksson, Niclas; Wojcik, Jerome; Reddy, Prasad Linga; Lima, Guadalupe; D’Alfonso, Sandra; Migliaresi, Sergio; Baca, Vicente; Orozco, Lorena; Witte, Torsten; Ortego-Centeno, Norberto; Abderrahim, Hadi; Pons-Estel, Bernardo A.; Gutiérrez, Carmen; Suárez, Ana; González-Escribano, Maria Francisca; Martin, Javier; Alarcón-Riquelme, Marta E.

    2013-01-01

    Objectives To confirm and define the genetic association of STAT4 and systemic lupus erythematosus, investigate the possibility of correlations with differential splicing and/or expression levels, and genetic interaction with IRF5. Methods 30 tag SNPs were genotyped in an independent set of Spanish cases and controls. SNPs surviving correction for multiple tests were genotyped in 5 new sets of cases and controls for replication. STAT4 cDNA was analyzed by 5’-RACE PCR and sequencing. Expression levels were measured by quantitative PCR. Results In the fine-mapping, four SNPs were significant after correction for multiple testing, with rs3821236 and rs3024866 as the strongest signals, followed by the previously associated rs7574865, and by rs1467199. Association was replicated in all cohorts. After conditional regression analyses, two major independent signals represented by SNPs rs3821236 and rs7574865, remained significant across the sets. These SNPs belong to separate haplotype blocks. High levels of STAT4 expression correlated with SNPs rs3821236, rs3024866 (both in the same haplotype block) and rs7574865 but not with other SNPs. We also detected transcription of alternative tissue-specific exons 1, indicating presence of tissue-specific promoters of potential importance in the expression of STAT4. No interaction with associated SNPs of IRF5 was observed using regression analysis. Conclusions These data confirm STAT4 as a susceptibility gene for SLE and suggest the presence of at least two functional variants affecting levels of STAT4. Our results also indicate that both genes STAT4 and IRF5 act additively to increase risk for SLE. PMID:19019891

  8. Properties of high calcium fly ash geopolymer pastes with Portland cement as an additive

    NASA Astrophysics Data System (ADS)

    Phoo-ngernkham, Tanakorn; Chindaprasirt, Prinya; Sata, Vanchai; Pangdaeng, Saengsuree; Sinsiri, Theerawat

    2013-02-01

    The effect of Portland cement (OPC) addition on the properties of high calcium fly ash geopolymer pastes was investigated in the paper. OPC partially replaced fly ash (FA) at the dosages of 0, 5%, 10%, and 15% by mass of binder. Sodium silicate (Na2SiO3) and sodium hydroxide (NaOH) solutions were used as the liquid portion in the mixture: NaOH 10 mol/L, Na2SiO3/NaOH with a mass ratio of 2.0, and alkaline liquid/binder (L/B) with a mass ratio of 0.6. The curing at 60°C for 24 h was used to accelerate the geopolymerization. The setting time of all fresh pastes, porosity, and compressive strength of the pastes at the stages of 1, 7, 28, and 90 d were tested. The elastic modulus and strain capacity of the pastes at the stage of 7 d were determined. It is revealed that the use of OPC as an additive to replace part of FA results in the decreases in the setting time, porosity, and strain capacity of the paste specimens, while the compressive strength and elastic modulus seem to increase.

  9. Enhanced high-solids anaerobic digestion of waste activated sludge by the addition of scrap iron.

    PubMed

    Zhang, Yaobin; Feng, Yinghong; Yu, Qilin; Xu, Zibin; Quan, Xie

    2014-05-01

    Anaerobic digestion of waste activated sludge usually requires pretreatment procedure to improve the bioavailability of sludge, which involves considerable energy and high expenditures. This study proposes a cost-effective method for enhanced anaerobic digestion of sludge without a pretreatment by directly adding iron into the digester. The results showed that addition of Fe(0) powder could enhance 14.46% methane yield, and Fe scrap (clean scrap) could further enhance methane yield (improving rate 21.28%) because the scrap has better mass transfer efficiency with sludge and liquid than Fe(0) powder. The scrap of Fe with rust (rusty scrap) could induce microbial Fe(III) reduction, which resulted in achieving the highest methane yield (improving rate 29.51%), and the reduction rate of volatile suspended solids (VSS) was also highest (48.27%) among Fe powder, clean scrap and rusty scrap. PCR-DGGE proved that the addition of rusty scrap could enhance diversity of acetobacteria and enrich iron-reducing bacteria to enhance degradation of complex substrates. PMID:24657762

  10. Enhanced high-solids anaerobic digestion of waste activated sludge by the addition of scrap iron.

    PubMed

    Zhang, Yaobin; Feng, Yinghong; Yu, Qilin; Xu, Zibin; Quan, Xie

    2014-05-01

    Anaerobic digestion of waste activated sludge usually requires pretreatment procedure to improve the bioavailability of sludge, which involves considerable energy and high expenditures. This study proposes a cost-effective method for enhanced anaerobic digestion of sludge without a pretreatment by directly adding iron into the digester. The results showed that addition of Fe(0) powder could enhance 14.46% methane yield, and Fe scrap (clean scrap) could further enhance methane yield (improving rate 21.28%) because the scrap has better mass transfer efficiency with sludge and liquid than Fe(0) powder. The scrap of Fe with rust (rusty scrap) could induce microbial Fe(III) reduction, which resulted in achieving the highest methane yield (improving rate 29.51%), and the reduction rate of volatile suspended solids (VSS) was also highest (48.27%) among Fe powder, clean scrap and rusty scrap. PCR-DGGE proved that the addition of rusty scrap could enhance diversity of acetobacteria and enrich iron-reducing bacteria to enhance degradation of complex substrates.

  11. [Simultaneous determination of various food additives by high performance liquid chromatography].

    PubMed

    Chen, Q C; Yu, W L; Wang, J

    2001-03-01

    A novel method is proposed for the simultaneous separation and determination of eight food additives, acesulfame potassium (AK), aspartame (ASP), benzoic acid (BA), caffeine (CA), saccharin sodium (SA), sorbic acid (SOR), theobromine (TB) and theophylline (TP) by reversed-phase high performance liquid chromatography. The separation was achieved within 23 min by using an Alltech Econosphere C18 column with 10 mmol/L NaH2PO4(pH 4.00)-acetonitrile (90:10, V/V) as mobile phase. The qualification and quantitation were accomplished by using a photodiode array detector. The detection limits (S/N = 3) for all analytes were below mg/L level. Under the experimental conditions, other common food additives and organic acids such as cyclamate, citric acid, malic acid, tartaric acid and ascorbic acid, did not interfere with the determination. The method has been successfully applied to the analysis of various foods as well as pharmaceutical preparation, and the average recoveries for real samples ranged from 78.5% to 107.2%.

  12. Reductive burning of high-yield spent pulping liquors by the addition of pulverized coal

    SciTech Connect

    Sell, N.J.; Norman, J.C. )

    1992-10-01

    This paper reports on the reductive burning of high-yield spent pulping liquors which can be accomplished by the addition of pulverized coal to increase the heat content and generate the proper reducing conditions. Samples from a 78%-yield sodium bisulfite pulping process employing a hardwood furnish were mixed with 10-50% pulveriized coal and burned at 950[degrees]C under reducing conditions in a box furnace. Even in these uncontrolled combustion conditions 76. 5% of the sulfur found in the soluble portion of the smelt was converted from lignousulfonates to useful sulfide ion. For the remainder of the sulfur, analyses determined it to be 19. 5% as sulfite ion, 3. 1% as thiosulfate ion, and 0.9% as sulfate ion.

  13. Bifunctional hairy silica nanoparticles as high-performance additives for lubricant

    PubMed Central

    Sui, Tianyi; Song, Baoyu; Wen, Yu-ho; Zhang, Feng

    2016-01-01

    Bifunctional hairy silica nanoparticles (BHSNs), which are silica nanoparticles covered with alkyl and amino organic chains, were prepared as high-performance additives for lubricants. Compared with hairy silica nanoparticles covered by a single type of organic chain, binary hairy silica nanoparticles exhibit the advantages of both types of organic chains, which exhibit excellent compatibility with lubricants and adsorbability to metal surfaces. Nanoparticles with different ratios of amino and alkyl ligands were investigated. In comparison to an untreated lubricant, BHSNs reduce the friction coefficient and wear scar diameter by 40% and 60%, respectively. The wear mechanism of BHSNs was investigated, and the protective and filling effect of the nanoparticles improved because of collaboration of amino and alkyl ligands. PMID:26936117

  14. Highly stereoselective, cobalt(III)-directed Mannich additions in water yielding α-methylamino acid products.

    PubMed

    Oerum, Stephanie; Krabbe, Peter; Schau-Magnussen, Magnus; Furbo, Søren; Bendix, Jesper; Hammershøi, Anders

    2013-01-01

    Highly stereoselective and rapid (<1 min) addition reactions to the imine double bond of 2-(methylimino)acetate complexes [L(4)Co(O(2)CCH=NCH(3))](2+) [L(4) = (en)(2) (7), (tren) (11)] were achieved in aqueous solution with nitromethane, ethyl 3-oxobutanoate or diethyl malonate. The molecular structures of two product complexes, rac-(Δ*-R(C)*-S(N)*)-[Co(en)(2)(O(2)CCH[CH(2)NO(2)]NHCH(3))]ZnCl(4) and rac-(Δ*-R(C)*-S(N)*)-[Co(en)(2)(O(2)CCH[CH(2)COCH(3)]NHCH(3))]ZnCl(4), were established by X-ray diffraction.

  15. Additive Manufacturing and High-Performance Computing: a Disruptive Latent Technology

    NASA Astrophysics Data System (ADS)

    Goodwin, Bruce

    2015-03-01

    This presentation will discuss the relationship between recent advances in Additive Manufacturing (AM) technology, High-Performance Computing (HPC) simulation and design capabilities, and related advances in Uncertainty Quantification (UQ), and then examines their impacts upon national and international security. The presentation surveys how AM accelerates the fabrication process, while HPC combined with UQ provides a fast track for the engineering design cycle. The combination of AM and HPC/UQ almost eliminates the engineering design and prototype iterative cycle, thereby dramatically reducing cost of production and time-to-market. These methods thereby present significant benefits for US national interests, both civilian and military, in an age of austerity. Finally, considering cyber security issues and the advent of the ``cloud,'' these disruptive, currently latent technologies may well enable proliferation and so challenge both nuclear and non-nuclear aspects of international security.

  16. Highly treated mine waters may require major ion addition before environmental release.

    PubMed

    Harford, Andrew J; Jones, David R; van Dam, Rick A

    2013-01-15

    Mining operations often use passive and/or active water treatments to improve water quality prior to environmental release. Key considerations in choosing a treatment process include the extent to which the water quality is actually improved, and the potential residual environmental risks of the release of such water. However, there are few published studies concerning the environmental impacts of treated waste waters. This study used toxicity identification evaluation (TIE) methods to quantify and identify the "toxic" constituents of a highly-treated water (distillate) produced by brine concentration of a mining process water. Exposure of five freshwater species (Chlorella sp., Lemna aequinoctialis, Hydra viridissima, Moinodaphnia macleayi and Mogurnda mogurnda) to a concentration range of the distillate (0, 25, 50 and 100%) found that it was toxic to H. viridissima (50-100% effect when exposed to 100% distillate). TIE tests demonstrated that the effect wasn't due to residual ammonia (~1 mg L(-1)N) or trace organics, and unlikely to be due to manganese (Mn; 130-230 μg L(-1)). Conversely, addition of 0.2 and 0.5 mg L(-1) calcium improved the growth rate of H. viridissima by 61 and 66%, respectively, while addition of calcium, sodium and potassium (0.5, 1.0 and 0.4 mg L(-1), respectively) to levels comparable to that in the local aquatic environment resulted in 100% recovery. Further assessment on the likelihood of residual metal toxicity indicated that Mn concentrations in the distillate were at levels that could inhibit the growth of H. viridissima. Ultimately, the results demonstrated that ion deficiency should be considered as a potential stressor in risk/impact assessments of the discharge of treated wastewaters, and these may need to be supplemented with the deficient ions to reduce environmental impacts. The findings have highlighted the need for water managers to consider the possibility of unintended environmental risks from the discharge of highly

  17. Highly treated mine waters may require major ion addition before environmental release.

    PubMed

    Harford, Andrew J; Jones, David R; van Dam, Rick A

    2013-01-15

    Mining operations often use passive and/or active water treatments to improve water quality prior to environmental release. Key considerations in choosing a treatment process include the extent to which the water quality is actually improved, and the potential residual environmental risks of the release of such water. However, there are few published studies concerning the environmental impacts of treated waste waters. This study used toxicity identification evaluation (TIE) methods to quantify and identify the "toxic" constituents of a highly-treated water (distillate) produced by brine concentration of a mining process water. Exposure of five freshwater species (Chlorella sp., Lemna aequinoctialis, Hydra viridissima, Moinodaphnia macleayi and Mogurnda mogurnda) to a concentration range of the distillate (0, 25, 50 and 100%) found that it was toxic to H. viridissima (50-100% effect when exposed to 100% distillate). TIE tests demonstrated that the effect wasn't due to residual ammonia (~1 mg L(-1)N) or trace organics, and unlikely to be due to manganese (Mn; 130-230 μg L(-1)). Conversely, addition of 0.2 and 0.5 mg L(-1) calcium improved the growth rate of H. viridissima by 61 and 66%, respectively, while addition of calcium, sodium and potassium (0.5, 1.0 and 0.4 mg L(-1), respectively) to levels comparable to that in the local aquatic environment resulted in 100% recovery. Further assessment on the likelihood of residual metal toxicity indicated that Mn concentrations in the distillate were at levels that could inhibit the growth of H. viridissima. Ultimately, the results demonstrated that ion deficiency should be considered as a potential stressor in risk/impact assessments of the discharge of treated wastewaters, and these may need to be supplemented with the deficient ions to reduce environmental impacts. The findings have highlighted the need for water managers to consider the possibility of unintended environmental risks from the discharge of highly

  18. A novel baculovirus-derived promoter with high activity in the baculovirus expression system.

    PubMed

    Martínez-Solís, María; Gómez-Sebastián, Silvia; Escribano, José M; Jakubowska, Agata K; Herrero, Salvador

    2016-01-01

    The baculovirus expression vector system (BEVS) has been widely used to produce a large number of recombinant proteins, and is becoming one of the most powerful, robust, and cost-effective systems for the production of eukaryotic proteins. Nevertheless, as in any other protein expression system, it is important to improve the production capabilities of this vector. The orf46 viral gene was identified among the most highly abundant sequences in the transcriptome of Spodoptera exigua larvae infected with its native baculovirus, the S. exigua multiple nucleopolyhedrovirus (SeMNPV). Different sequences upstream of the orf46 gene were cloned, and their promoter activities were tested by the expression of the GFP reporter gene using the Autographa californica nucleopolyhedrovirus (AcMNPV) vector system in different insect cell lines (Sf21, Se301, and Hi5) and in larvae from S. exigua and Trichoplusia ni. The strongest promoter activity was defined by a 120 nt sequence upstream of the ATG start codon for the orf46 gene. On average, GFP expression under this new promoter was more than two fold higher than the expression obtained with the standard polyhedrin (polh) promoter. Additionally, the orf46 promoter was also tested in combination with the polh promoter, revealing an additive effect over the polh promoter activity. In conclusion, this new characterized promoter represents an excellent alternative to the most commonly used baculovirus promoters for the efficient expression of recombinant proteins using the BEVS. PMID:27375973

  19. A novel baculovirus-derived promoter with high activity in the baculovirus expression system

    PubMed Central

    Martínez-Solís, María; Gómez-Sebastián, Silvia; Escribano, José M.; Jakubowska, Agata K.

    2016-01-01

    The baculovirus expression vector system (BEVS) has been widely used to produce a large number of recombinant proteins, and is becoming one of the most powerful, robust, and cost-effective systems for the production of eukaryotic proteins. Nevertheless, as in any other protein expression system, it is important to improve the production capabilities of this vector. The orf46 viral gene was identified among the most highly abundant sequences in the transcriptome of Spodoptera exigua larvae infected with its native baculovirus, the S. exigua multiple nucleopolyhedrovirus (SeMNPV). Different sequences upstream of the orf46 gene were cloned, and their promoter activities were tested by the expression of the GFP reporter gene using the Autographa californica nucleopolyhedrovirus (AcMNPV) vector system in different insect cell lines (Sf21, Se301, and Hi5) and in larvae from S. exigua and Trichoplusia ni. The strongest promoter activity was defined by a 120 nt sequence upstream of the ATG start codon for the orf46 gene. On average, GFP expression under this new promoter was more than two fold higher than the expression obtained with the standard polyhedrin (polh) promoter. Additionally, the orf46 promoter was also tested in combination with the polh promoter, revealing an additive effect over the polh promoter activity. In conclusion, this new characterized promoter represents an excellent alternative to the most commonly used baculovirus promoters for the efficient expression of recombinant proteins using the BEVS. PMID:27375973

  20. A novel baculovirus-derived promoter with high activity in the baculovirus expression system.

    PubMed

    Martínez-Solís, María; Gómez-Sebastián, Silvia; Escribano, José M; Jakubowska, Agata K; Herrero, Salvador

    2016-01-01

    The baculovirus expression vector system (BEVS) has been widely used to produce a large number of recombinant proteins, and is becoming one of the most powerful, robust, and cost-effective systems for the production of eukaryotic proteins. Nevertheless, as in any other protein expression system, it is important to improve the production capabilities of this vector. The orf46 viral gene was identified among the most highly abundant sequences in the transcriptome of Spodoptera exigua larvae infected with its native baculovirus, the S. exigua multiple nucleopolyhedrovirus (SeMNPV). Different sequences upstream of the orf46 gene were cloned, and their promoter activities were tested by the expression of the GFP reporter gene using the Autographa californica nucleopolyhedrovirus (AcMNPV) vector system in different insect cell lines (Sf21, Se301, and Hi5) and in larvae from S. exigua and Trichoplusia ni. The strongest promoter activity was defined by a 120 nt sequence upstream of the ATG start codon for the orf46 gene. On average, GFP expression under this new promoter was more than two fold higher than the expression obtained with the standard polyhedrin (polh) promoter. Additionally, the orf46 promoter was also tested in combination with the polh promoter, revealing an additive effect over the polh promoter activity. In conclusion, this new characterized promoter represents an excellent alternative to the most commonly used baculovirus promoters for the efficient expression of recombinant proteins using the BEVS.

  1. Yeast-containing feed additive alters gene expression profiles associated with innate immunity in whole blood of a rodent model.

    PubMed

    Branson, Jennifer A; McLean, Derek J; Forsberg, Neil E; Bobe, Gerd

    2016-05-01

    Feeding a yeast-containing additive (YCA; OmniGen-AF) improves immune responses in ruminant livestock and reduces subsequent production losses. The objective was to identify molecular pathways by which dietary YCA may modify immune responses using a rodent model. Thirty-seven healthy, unchallenged CD rats received a diet containing 0 (control; n = 5, only 28 d), 0.5% (n = 15) or 1% (n = 17) YCA for 7 (n = 4/group), 14 (n = 3 or 4/group), 21 (n = 3 or 4/group) or 28 (n = 5/group) d. At the end of the feeding periods, whole blood was collected and the isolated RNA was analyzed for the expression of 84 genes involved in innate and cell-mediated adaptive immune responses. Three bacterial pattern recognition receptors TLR1 (0.5%: + 2.01; 1%: + 2.38), TLR6 (0.5%: + 2.11; 1%: + 2.34) and NOD2 (0.5%: + 2.32; 1%: + 2.23), two APC surface receptors CD1D1 (0.5%: + 1.75; 1%: + 2.33) and CD80 (0.5%: +2.45; 1%: +3.00), and the cell signaling molecule MAPK8 (0.5%: +1.87; 1%: +2.35) were significantly up-regulated by YCA at both inclusion rates. In conclusion, feeding YCA may potentially increase recognition and responses to bacterial pathogens and T-cell activation and differentiation and thereby maintain health and prevent production losses. PMID:27033362

  2. Nonparametric Independence Screening in Sparse Ultra-High Dimensional Additive Models.

    PubMed

    Fan, Jianqing; Feng, Yang; Song, Rui

    2011-06-01

    A variable screening procedure via correlation learning was proposed in Fan and Lv (2008) to reduce dimensionality in sparse ultra-high dimensional models. Even when the true model is linear, the marginal regression can be highly nonlinear. To address this issue, we further extend the correlation learning to marginal nonparametric learning. Our nonparametric independence screening is called NIS, a specific member of the sure independence screening. Several closely related variable screening procedures are proposed. Under general nonparametric models, it is shown that under some mild technical conditions, the proposed independence screening methods enjoy a sure screening property. The extent to which the dimensionality can be reduced by independence screening is also explicitly quantified. As a methodological extension, a data-driven thresholding and an iterative nonparametric independence screening (INIS) are also proposed to enhance the finite sample performance for fitting sparse additive models. The simulation results and a real data analysis demonstrate that the proposed procedure works well with moderate sample size and large dimension and performs better than competing methods.

  3. Effects of Zn additions to highly magnetoelastic FeGa alloys

    SciTech Connect

    Lograsso, Thomas A.; Jones, Nicholas J.; Wun-Fogle, Marilyn; Restorff, James B.; Schlagel, Deborah L.; Petculescu, Gabriela; Clark, Arthur E.; Hathaway, Kristl B.

    2015-05-07

    Fe{sub 1−x}M{sub x} (M = Ga, Ge, Si, Al, Mo and x ∼ 0.18) alloys offer an extraordinary combination of magnetoelasticity and mechanical properties. They are rare-earth-free, can be processed using conventional deformation techniques, have high magnetic permeability, low hysteresis, and low magnetic saturation fields, making them attractive for device applications such as actuators and energy harvesters. Starting with Fe-Ga as a reference and using a rigid-band-filling argument, Zhang et al. predicted that lowering the Fermi level by reducing the total number of electrons could enhance magnetoelasticity. To provide a direct experimental validation for Zhang's hypothesis, elemental additions with lower-than-Ga valence are needed. Of the possible candidates, only Be and Zn have sufficient solubility. Single crystals of bcc Fe-Ga-Zn have been grown with up to 4.6 at. % Zn in a Bridgman furnace under elevated pressure (15 bars) in order to overcome the high vapor pressure of Zn and obtain homogeneous crystals. Single-crystal measurements of magnetostriction and elastic constants allow for the direct comparison of the magnetoelastic coupling constants of Fe-Ga-Zn with those of other magnetoelastic alloys in its class. The partial substitution of Ga with Zn yields values for the magnetoelastic coupling factor, −b{sub 1}, comparable to those of the binary Fe-Ga alloy.

  4. Development of magnetodielectric materials to be used in additive manufacturing processes for high-frequency applications

    NASA Astrophysics Data System (ADS)

    Parsons, Paul Emerson, II

    Electrical devices for very-high frequency (VHF, 0.03 -- 0.3 GHz) and ultra-high frequency (UHF, 0.3 -- 3.0 GHz) are commonly used for communications. However, the wavelengths, lambda, of these frequency bands correspond to lengths between 10 and 0.1 m, resulting in prohibitively large devices. Materials with an index of refraction, n, greater than 1 can be used to effectively shrink these devices by a factor of 1/ n. In this thesis, magnetodielectric materials (MDM), where n ≥1, have been made to be used in additive manufacturing processes with strict particle size requirements and were developed using various methods, such as polyol reduction and conventional ceramic solid state processing. These materials were characterized using x-ray diffraction (XRD), scanning and transmission electron microscopy (SEM and TEM), vibrating sample magnetometry (VSM), to determine their crystalline, physical, and direct current (DC) magnetization properties. The techniques used to synthesize the MDM yielded particles that were chemically similar, but had drastically different physical properties which heavily influences their high-frequency electromagnetic properties. These materials were then uniformly dispersed into a non-conducting medium, such as a low-electrical loss polymer or resin, and formed into composite samples with variable volumetric loading. These composite samples were measured using several techniques to characterize the frequency-dependent electromagnetic (EM) properties, such as relative permeability, relative permittivity, and their respective losses. Finite element method (FEM) simulations were performed using these MDM-composites to design a spiral antenna to be used at approximately 585 MHz.

  5. Hierarchical random additive process and logarithmic scaling of generalized high order, two-point correlations in turbulent boundary layer flow

    NASA Astrophysics Data System (ADS)

    Yang, X. I. A.; Marusic, I.; Meneveau, C.

    2016-06-01

    Townsend [Townsend, The Structure of Turbulent Shear Flow (Cambridge University Press, Cambridge, UK, 1976)] hypothesized that the logarithmic region in high-Reynolds-number wall-bounded flows consists of space-filling, self-similar attached eddies. Invoking this hypothesis, we express streamwise velocity fluctuations in the inertial layer in high-Reynolds-number wall-bounded flows as a hierarchical random additive process (HRAP): uz+=∑i=1Nzai . Here u is the streamwise velocity fluctuation, + indicates normalization in wall units, z is the wall normal distance, and ai's are independently, identically distributed random additives, each of which is associated with an attached eddy in the wall-attached hierarchy. The number of random additives is Nz˜ln(δ /z ) where δ is the boundary layer thickness and ln is natural log. Due to its simplified structure, such a process leads to predictions of the scaling behaviors for various turbulence statistics in the logarithmic layer. Besides reproducing known logarithmic scaling of moments, structure functions, and correlation function [" close="]3/2 uz(x ) uz(x +r ) >, new logarithmic laws in two-point statistics such as uz4(x ) > 1 /2, 1/3, etc. can be derived using the HRAP formalism. Supporting empirical evidence for the logarithmic scaling in such statistics is found from the Melbourne High Reynolds Number Boundary Layer Wind Tunnel measurements. We also show that, at high Reynolds numbers, the above mentioned new logarithmic laws can be derived by assuming the arrival of an attached eddy at a generic point in the flow field to be a Poisson process [Woodcock and Marusic, Phys. Fluids 27, 015104 (2015), 10.1063/1.4905301]. Taken together, the results provide new evidence supporting the essential ingredients of the attached eddy hypothesis to describe streamwise velocity fluctuations of large, momentum transporting eddies in wall-bounded turbulence, while

  6. Can "normal" protein expression ranges be estimated with high-throughput proteomics?

    PubMed

    Higdon, Roger; Kolker, Eugene

    2015-06-01

    Although biological science discovery often involves comparing conditions to a normal state, in proteomics little is actually known about normal. Two Human Proteome studies featured in Nature offer new insights into protein expression and an opportunity to assess how high-throughput proteomics measures normal protein ranges. We use data from these studies to estimate technical and biological variability in protein expression and compare them to other expression data sets from normal tissue. Results show that measured protein expression across same-tissue replicates vary by ±4- to 10-fold for most proteins. Coefficients of variation (CV) for protein expression measurements range from 62% to 117% across different tissue experiments; however, adjusting for technical variation reduced this variability by as much as 50%. In addition, the CV could also be reduced by limiting comparisons to proteins with at least 3 or more unique peptide identifications as the CV was on average 33% lower than for proteins with 2 or fewer peptide identifications. We also selected 13 housekeeping proteins and genes that were expressed across all tissues with low variability to determine their utility as a reference set for normalization and comparative purposes. These results present the first step toward estimating normal protein ranges by determining the variability in expression measurements through combining publicly available data. They support an approach that combines standard protocols with replicates of normal tissues to estimate normal protein ranges for large numbers of proteins and tissues. This would be a tremendous resource for normal cellular physiology and comparisons of proteomics studies.

  7. Twister ribozymes as highly versatile expression platforms for artificial riboswitches

    PubMed Central

    Felletti, Michele; Stifel, Julia; Wurmthaler, Lena A.; Geiger, Sophie; Hartig, Jörg S.

    2016-01-01

    The utilization of ribozyme-based synthetic switches in biotechnology has many advantages such as an increased robustness due to in cis regulation, small coding space and a high degree of modularity. The report of small endonucleolytic twister ribozymes provides new opportunities for the development of advanced tools for engineering synthetic genetic switches. Here we show that the twister ribozyme is distinguished as an outstandingly flexible expression platform, which in conjugation with three different aptamer domains, enables the construction of many different one- and two-input regulators of gene expression in both bacteria and yeast. Besides important implications in biotechnology and synthetic biology, the observed versatility in artificial genetic control set-ups hints at possible natural roles of this widespread ribozyme class. PMID:27670347

  8. High magnetic field induced changes of gene expression in arabidopsis

    PubMed Central

    Paul, Anna-Lisa; Ferl, Robert J; Meisel, Mark W

    2006-01-01

    Background High magnetic fields are becoming increasingly prevalent components of non-invasive, biomedical imaging tools (such as MRI), thus, an understanding of the molecular impacts associated with these field strengths in biological systems is of central importance. The biological impact of magnetic field strengths up to 30 Tesla were investigated in this study through the use of transgenic Arabidopsis plants engineered with a stress response gene consisting of the alcohol dehydrogenase (Adh) gene promoter driving the β-glucuronidase (GUS) gene reporter. Methods Magnetic field induced Adh/GUS activity was evaluated with histochemical staining to assess tissue specific expression and distribution, and with quantitative, spectrofluometric assays to measure degree of activation. The evaluation of global changes in the Arabidopsis genome in response to exposure to high magnetic fields was facilitated with Affymetrix Gene Chip microarrays. Quantitative analyses of gene expression were performed with quantitative real-time polymerase-chain-reaction (qRT-PCR). Results Field strengths in excess of about 15 Tesla induce expression of the Adh/GUS transgene in the roots and leaves. From the microarray analyses that surveyed 8000 genes, 114 genes were differentially expressed to a degree greater than 2.5 fold over the control. These results were quantitatively corroborated by qRT-PCR examination of 4 of the 114 genes. Conclusion The data suggest that magnetic fields in excess of 15 Tesla have far-reaching effect on the genome. The wide-spread induction of stress-related genes and transcription factors, and a depression of genes associated with cell wall metabolism, are prominent examples. The roles of magnetic field orientation of macromolecules and magnetophoretic effects are discussed as possible factors that contribute to the mounting of this response. PMID:17187667

  9. Express Primer Tool for high-throughput gene cloning and expression

    2002-12-01

    A tool to assist in the design of primers for DNA amplification. The Express Primer web-based tool generates primer sequences specifically for the generation of expression clones for both lab scale and high-throughput projects. The application is designed not only to allow the user complete flexibility to specify primer design parameters but also to minimize the amount of manual intervention needed to generate a large number of primers for simultaneous amplification of multiple target genes.more » The Express Primer Tool enables the user to specify various experimental parameters (e.g. optimal Tm, Tm range, maximum Tm difference) for single or multiple candidate sequence(s) in FASTA format input as a flat text (ASCII) file. The application generates condidate primers, selects optimal primer pairs, and writes the forward and reverse primers pairs to an Excel file that is suitable for electronic submission to a synthesis facility. The program parameters emphasize high-throughput but allow for target atrition at various stages of the project.« less

  10. Continuous Digital Light Processing (cDLP): Highly Accurate Additive Manufacturing of Tissue Engineered Bone Scaffolds.

    PubMed

    Dean, David; Jonathan, Wallace; Siblani, Ali; Wang, Martha O; Kim, Kyobum; Mikos, Antonios G; Fisher, John P

    2012-03-01

    Highly accurate rendering of the external and internal geometry of bone tissue engineering scaffolds effects fit at the defect site, loading of internal pore spaces with cells, bioreactor-delivered nutrient and growth factor circulation, and scaffold resorption. It may be necessary to render resorbable polymer scaffolds with 50 μm or less accuracy to achieve these goals. This level of accuracy is available using Continuous Digital Light processing (cDLP) which utilizes a DLP(®) (Texas Instruments, Dallas, TX) chip. One such additive manufacturing device is the envisionTEC (Ferndale, MI) Perfactory(®). To use cDLP we integrate a photo-crosslinkable polymer, a photo-initiator, and a biocompatible dye. The dye attenuates light, thereby limiting the depth of polymerization. In this study we fabricated scaffolds using the well-studied resorbable polymer, poly(propylene fumarate) (PPF), titanium dioxide (TiO(2)) as a dye, Irgacure(®) 819 (BASF [Ciba], Florham Park, NJ) as an initiator, and diethyl fumarate as a solvent to control viscosity. PMID:23066427

  11. Estimation and Inference in Generalized Additive Coefficient Models for Nonlinear Interactions with High-Dimensional Covariates

    PubMed Central

    Shujie, MA; Carroll, Raymond J.; Liang, Hua; Xu, Shizhong

    2015-01-01

    In the low-dimensional case, the generalized additive coefficient model (GACM) proposed by Xue and Yang [Statist. Sinica 16 (2006) 1423–1446] has been demonstrated to be a powerful tool for studying nonlinear interaction effects of variables. In this paper, we propose estimation and inference procedures for the GACM when the dimension of the variables is high. Specifically, we propose a groupwise penalization based procedure to distinguish significant covariates for the “large p small n” setting. The procedure is shown to be consistent for model structure identification. Further, we construct simultaneous confidence bands for the coefficient functions in the selected model based on a refined two-step spline estimator. We also discuss how to choose the tuning parameters. To estimate the standard deviation of the functional estimator, we adopt the smoothed bootstrap method. We conduct simulation experiments to evaluate the numerical performance of the proposed methods and analyze an obesity data set from a genome-wide association study as an illustration. PMID:26412908

  12. Additions and Improvements to the FLASH Code for Simulating High Energy Density Physics Experiments

    NASA Astrophysics Data System (ADS)

    Lamb, D. Q.; Daley, C.; Dubey, A.; Fatenejad, M.; Flocke, N.; Graziani, C.; Lee, D.; Tzeferacos, P.; Weide, K.

    2015-11-01

    FLASH is an open source, finite-volume Eulerian, spatially adaptive radiation hydrodynamics and magnetohydrodynamics code that incorporates capabilities for a broad range of physical processes, performs well on a wide range of computer architectures, and has a broad user base. Extensive capabilities have been added to FLASH to make it an open toolset for the academic high energy density physics (HEDP) community. We summarize these capabilities, with particular emphasis on recent additions and improvements. These include advancements in the optical ray tracing laser package, with methods such as bi-cubic 2D and tri-cubic 3D interpolation of electron number density, adaptive stepping and 2nd-, 3rd-, and 4th-order Runge-Kutta integration methods. Moreover, we showcase the simulated magnetic field diagnostic capabilities of the code, including induction coils, Faraday rotation, and proton radiography. We also describe several collaborations with the National Laboratories and the academic community in which FLASH has been used to simulate HEDP experiments. This work was supported in part at the University of Chicago by the DOE NNSA ASC through the Argonne Institute for Computing in Science under field work proposal 57789; and the NSF under grant PHY-0903997.

  13. Continuous Digital Light Processing (cDLP): Highly Accurate Additive Manufacturing of Tissue Engineered Bone Scaffolds

    PubMed Central

    Dean, David; Wallace, Jonathan; Siblani, Ali; Wang, Martha O.; Kim, Kyobum; Mikos, Antonios G.; Fisher, John P.

    2012-01-01

    Highly accurate rendering of the external and internal geometry of bone tissue engineering scaffolds effects fit at the defect site, loading of internal pore spaces with cells, bioreactor-delivered nutrient and growth factor circulation, and scaffold resorption. It may be necessary to render resorbable polymer scaffolds with 50 μm or less accuracy to achieve these goals. This level of accuracy is available using Continuous Digital Light processing (cDLP) which utilizes a DLP® (Texas Instruments, Dallas, TX) chip. One such additive manufacturing device is the envisionTEC (Ferndale, MI) Perfactory®. To use cDLP we integrate a photo-crosslinkable polymer, a photo-initiator, and a biocompatible dye. The dye attenuates light, thereby limiting the depth of polymerization. In this study we fabricated scaffolds using the well-studied resorbable polymer, poly(propylene fumarate) (PPF), titanium dioxide (TiO2) as a dye, Irgacure® 819 (BASF [Ciba], Florham Park, NJ) as an initiator, and diethyl fumarate as a solvent to control viscosity. PMID:23066427

  14. High expression of cellular retinol binding protein-1 in lung adenocarcinoma is associated with poor prognosis

    PubMed Central

    Doldo, Elena; Costanza, Gaetana; Ferlosio, Amedeo; Pompeo, Eugenio; Agostinelli, Sara; Bellezza, Guido; Mazzaglia, Donatella; Giunta, Alessandro; Sidoni, Angelo; Orlandi, Augusto

    2015-01-01

    Purpose Adenocarcinoma, the most common non-small cell lung cancer is a leading cause of death worldwide, with a low overall survival (OS) despite increasing attempts to achieve an early diagnosis and accomplish surgical and multimodality treatment strategies. Cellular retinol binding protein-1 (CRBP-1) regulates retinol bioavailability and cell differentiation, but its role in lung cancerogenesis remains uncertain. Experimental design CRBP-1 expression, clinical outcome and other prognostic factors were investigated in 167 lung adenocarcinoma patients. CRBP-1 expression was evaluated by immunohistochemistry of tissue microarray sections, gene copy number analysis and tumor methylation specific PCR. Effects of CRBP-1 expression on proliferation/apoptosis gene array, protein and transcripts were investigated in transfected A549 lung adenocarcinoma cells. Results CRBP-1High expression was observed in 62.3% of adenocarcinomas and correlated with increased tumor grade and reduced OS as an independent prognostic factor. CRBP-1 gene copy gain also associated with tumor CRBP-1High status and dedifferentiation. CRBP-1-transfected (CRBP-1+) A549 grew more than CRBP-1− A549 cells. At >1μM concentrations, all trans-retinoic acid and retinol reduced viability more in CRBP-1+ than in CRBP-1− A549 cells. CRBP-1+ A549 cells showed up-regulated RARα/ RXRα and proliferative and transcriptional genes including pAkt, pEGFR, pErk1/2, creb1 and c-jun, whereas RARβ and p53 were strongly down-regulated; pAkt/pErk/ pEGFR inhibitors counteracted proliferative advantage and increased RARα/RXRα, c-jun and CD44 expression in CRBP-1+ A549 cells. Conclusion CRBP-1High expression in lung adenocarcinoma correlated with increased tumor grade and reduced OS, likely through increased Akt/Erk/EGFR-mediated cell proliferation and differentiation. CRBP-1High expression can be considered an additional marker of poor prognosis in lung adenocarcinoma patients. PMID:26807202

  15. Carbon flux from plants to soil microbes is highly sensitive to nitrogen addition and biochar amendment

    NASA Astrophysics Data System (ADS)

    Kaiser, C.; Solaiman, Z. M.; Kilburn, M. R.; Clode, P. L.; Fuchslueger, L.; Koranda, M.; Murphy, D. V.

    2012-04-01

    material, microbial biomass and dissolved organic matter by IRMS, 13C and 15N in plant roots cells and intraradical mycorrhizal hyphae by NanoSims). Our results show that (1) C assimilated by plants was delivered within 4 hours to the soil microbial community both via roots and the mycorrhizal network (2) N addition during the labeling period strongly and rapidly increased the 13C flux of recently assimilated carbohydrates to the soil microbial biomass (3) the effect of N addition was not as rapid but was of the same magnitude when N was delivered to the plant exclusively by mycorrhizal hyphae as compared to taken up by roots (4) soils which had been amended with biochar (which were characterized by an increased abundance of mycorrhizal fungi) also showed a significant increase of C flux from plants to the soil. We conclude that plant belowground C allocation is highly sensitive to alterations of microbial community structure and nutritional status in the soil. Moreover, our results indicate that plants respond rapidly (within hours) to changing soil N availability by altering the rate of C transported belowground. Our results emphasise the ecological significance of plant-belowground interactions for ecosystem C cycling.

  16. High level expression of organophosphorus hydrolase in Pichia pastoris by multicopy ophcM assembly.

    PubMed

    Shen, Wei; Shu, Min; Ma, Lixin; Ni, Hong; Yan, Hong

    2016-03-01

    The residues of organophosphorus pesticides bring serious impact on the environmental safety and people's health. Biodegradation of organophosphorus pesticides is recognized as an ideal method. An organophosphorus hydrolase (OPHCM) from Pseudomonas pseudoalcaligenes was synthesized and expressed in Pichia pastoris. The yield reached approximately 470 mg/l after a 6-d induction in shake flasks. To improve the enzyme production, we describe a novel approach to express OPHCM efficiently with a biobrick assembly method in vitro. Four recombinant plasmids containing 1-4 copies of ophcM-expressing cassettes were constructed and transformed into P. pastoris. Increasing the copy number of ophcM gene enhanced the expression level of OPHCM. The maximum yield and specific activity in P. pastoris harboring two-copy tandem ophcM-expressing cassettes reached 610 mg/l after a 6-d induction in shake flasks and 7.8 g/l in high-density fermentation with specific activity of 13.7 U/mg. The optimum pH and temperature of the recombinant OPHCM activity were 11.0 and 50 °C, respectively. In addition, the enzyme activity of recombinant OPHCM enhanced 57.6% and 30.1% in the presence of 1 mM Cd(2+) and 5% glycerol, respectively. The high expression and good properties of recombinant OPHCM provide an effective solution to solve the pollution of organophosphorus pesticides in the environment. Moreover, the approach for generating multicopy gene expressing vectors here will benefit the study for enhancing the expression level of genes of interest. PMID:26611611

  17. Microarray Analysis of Gene Expression in Saccharomyces cerevisiae kap108Δ Mutants upon Addition of Oxidative Stress.

    PubMed

    Belanger, Kenneth D; Larson, Nathaniel; Kahn, Jonathan; Tkachev, Dmitry; Ay, Ahmet

    2016-01-01

    Protein transport between the nucleus and cytoplasm of eukaryotic cells is tightly regulated, providing a mechanism for controlling intracellular localization of proteins, and regulating gene expression. In this study, we have investigated the importance of nucleocytoplasmic transport mediated by the karyopherin Kap108 in regulating cellular responses to oxidative stress in Saccharomyces cerevisiae We carried out microarray analyses on wild-type and kap108 mutant cells grown under normal conditions, shortly after introduction of oxidative stress, after 1 hr of oxidative stress, and 1 hr after oxidative stress was removed. We observe more than 500 genes that undergo a 40% or greater change in differential expression between wild-type and kap108Δ cells under at least one of these conditions. Genes undergoing changes in expression can be categorized in two general groups: 1) those that are differentially expressed between wild-type and kap108Δ cells, no matter the oxidative stress conditions; and 2) those that have patterns of response dependent upon both the absence of Kap108, and introduction or removal of oxidative stress. Gene ontology analysis reveals that, among the genes whose expression is reduced in the absence of Kap108 are those involved in stress response and intracellular transport, while those overexpressed are largely involved in mating and pheromone response. We also identified 25 clusters of genes that undergo similar patterns of change in gene expression when oxidative stresses are added and subsequently removed, including genes involved in stress response, oxidation-reduction processing, iron homeostasis, ascospore wall assembly, transmembrane transport, and cell fusion during mating. These data suggest that Kap108 is important for regulating expression of genes involved in a variety of specific cell functions.

  18. The Wnt and Delta-Notch signalling pathways interact to direct pair-rule gene expression via caudal during segment addition in the spider Parasteatoda tepidariorum.

    PubMed

    Schönauer, Anna; Paese, Christian L B; Hilbrant, Maarten; Leite, Daniel J; Schwager, Evelyn E; Feitosa, Natália Martins; Eibner, Cornelius; Damen, Wim G M; McGregor, Alistair P

    2016-07-01

    In short-germ arthropods, posterior segments are added sequentially from a segment addition zone (SAZ) during embryogenesis. Studies in spiders such as Parasteatoda tepidariorum have provided insights into the gene regulatory network (GRN) underlying segment addition, and revealed that Wnt8 is required for dynamic Delta (Dl) expression associated with the formation of new segments. However, it remains unclear how these pathways interact during SAZ formation and segment addition. Here, we show that Delta-Notch signalling is required for Wnt8 expression in posterior SAZ cells, but represses the expression of this Wnt gene in anterior SAZ cells. We also found that these two signalling pathways are required for the expression of the spider orthologues of even-skipped (eve) and runt-1 (run-1), at least in part via caudal (cad). Moreover, it appears that dynamic expression of eve in this spider does not require a feedback loop with run-1, as is found in the pair-rule circuit of the beetle Tribolium Taken together, our results suggest that the development of posterior segments in Parasteatoda is directed by dynamic interactions between Wnt8 and Delta-Notch signalling that are read out by cad, which is necessary but probably not sufficient to regulate the expression of eve and run-1 Our study therefore provides new insights towards better understanding the evolution and developmental regulation of segmentation in other arthropods, including insects. PMID:27287802

  19. The Wnt and Delta-Notch signalling pathways interact to direct pair-rule gene expression via caudal during segment addition in the spider Parasteatoda tepidariorum.

    PubMed

    Schönauer, Anna; Paese, Christian L B; Hilbrant, Maarten; Leite, Daniel J; Schwager, Evelyn E; Feitosa, Natália Martins; Eibner, Cornelius; Damen, Wim G M; McGregor, Alistair P

    2016-07-01

    In short-germ arthropods, posterior segments are added sequentially from a segment addition zone (SAZ) during embryogenesis. Studies in spiders such as Parasteatoda tepidariorum have provided insights into the gene regulatory network (GRN) underlying segment addition, and revealed that Wnt8 is required for dynamic Delta (Dl) expression associated with the formation of new segments. However, it remains unclear how these pathways interact during SAZ formation and segment addition. Here, we show that Delta-Notch signalling is required for Wnt8 expression in posterior SAZ cells, but represses the expression of this Wnt gene in anterior SAZ cells. We also found that these two signalling pathways are required for the expression of the spider orthologues of even-skipped (eve) and runt-1 (run-1), at least in part via caudal (cad). Moreover, it appears that dynamic expression of eve in this spider does not require a feedback loop with run-1, as is found in the pair-rule circuit of the beetle Tribolium Taken together, our results suggest that the development of posterior segments in Parasteatoda is directed by dynamic interactions between Wnt8 and Delta-Notch signalling that are read out by cad, which is necessary but probably not sufficient to regulate the expression of eve and run-1 Our study therefore provides new insights towards better understanding the evolution and developmental regulation of segmentation in other arthropods, including insects.

  20. High-Density Lipoprotein Prevents Endoplasmic Reticulum Stress-Induced Downregulation of Liver LOX-1 Expression

    PubMed Central

    Hong, Dan; Li, Ling-Fang; Gao, Hai-Chao; Wang, Xiang; Li, Chuan-Chang; Luo, Ying; Bai, Yong-Ping; Zhang, Guo-Gang

    2015-01-01

    Lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) is a specific cell-surface receptor for oxidized-low-density lipoprotein (ox-LDL). The impact of high-density lipoprotein (HDL) on endoplasmic reticulum (ER) stress-mediated alteration of the LOX-1 level in hepatocytes remains unclear. We aimed to investigate the impact on LOX-1 expression by tunicamycin (TM)-induced ER stress and to determine the effect of HDL on TM-affected LOX-1 expression in hepatic L02 cells. Overexpression or silencing of related cellular genes was conducted in TM-treated cells. mRNA expression was evaluated using real-time polymerase chain reaction (PCR). Protein expression was analyzed by western blot and immunocytochemistry. Lipid uptake was examined by DiI-ox-LDL, followed by flow cytometric analysis. The results showed that TM induced the upregulation of ER chaperone GRP78, downregulation of LOX-1 expression, and lipid uptake. Knock down of IRE1 or XBP-1 effectively restored LOX-1 expression and improved lipid uptake in TM-treated cells. HDL treatment prevented the negative impact on LOX-1 expression and lipid uptake induced by TM. Additionally, 1–10 μg/mL HDL significantly reduced the GRP78, IRE1, and XBP-1 expression levels in TM-treated cells. Our findings reveal that HDL could prevent the TM-induced reduction of LOX-1 expression via inhibiting the IRE1/XBP-1 pathway, suggesting a new mechanism for beneficial roles of HDL in improving lipid metabolism. PMID:25923692

  1. High-Density Lipoprotein Prevents Endoplasmic Reticulum Stress-Induced Downregulation of Liver LOX-1 Expression.

    PubMed

    Hong, Dan; Li, Ling-Fang; Gao, Hai-Chao; Wang, Xiang; Li, Chuan-Chang; Luo, Ying; Bai, Yong-Ping; Zhang, Guo-Gang

    2015-01-01

    Lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) is a specific cell-surface receptor for oxidized-low-density lipoprotein (ox-LDL). The impact of high-density lipoprotein (HDL) on endoplasmic reticulum (ER) stress-mediated alteration of the LOX-1 level in hepatocytes remains unclear. We aimed to investigate the impact on LOX-1 expression by tunicamycin (TM)-induced ER stress and to determine the effect of HDL on TM-affected LOX-1 expression in hepatic L02 cells. Overexpression or silencing of related cellular genes was conducted in TM-treated cells. mRNA expression was evaluated using real-time polymerase chain reaction (PCR). Protein expression was analyzed by western blot and immunocytochemistry. Lipid uptake was examined by DiI-ox-LDL, followed by flow cytometric analysis. The results showed that TM induced the upregulation of ER chaperone GRP78, downregulation of LOX-1 expression, and lipid uptake. Knock down of IRE1 or XBP-1 effectively restored LOX-1 expression and improved lipid uptake in TM-treated cells. HDL treatment prevented the negative impact on LOX-1 expression and lipid uptake induced by TM. Additionally, 1-10 μg/mL HDL significantly reduced the GRP78, IRE1, and XBP-1 expression levels in TM-treated cells. Our findings reveal that HDL could prevent the TM-induced reduction of LOX-1 expression via inhibiting the IRE1/XBP-1 pathway, suggesting a new mechanism for beneficial roles of HDL in improving lipid metabolism. PMID:25923692

  2. Numerical Simulation of High Drag Reduction in a Turbulent Channel Flow with Polymer Additives

    NASA Technical Reports Server (NTRS)

    Dubief, Yves

    2003-01-01

    The addition of small amounts of long chain polymer molecules to wall-bounded flows can lead to dramatic drag reduction. Although this phenomenon has been known for about fifty years, the action of the polymers and its effect on turbulent structures are still unclear. Detailed experiments have characterized two distinct regimes (Warholic et al. 1999), which are referred to as low drag reduction (LDR) and high drag reduction (HDR). The first regime exhibits similar statistical trends as Newtonian flow: the log-law region of the mean velocity profile remains parallel to that of the Newtonian ow but its lower bound moves away from the wall and the upward shift of the log-region is a function of drag reduction, DR. Although streamwise fluctuations are increased and transverse ones are reduced, the shape of the rms velocity profiles is not qualitatively modified. At higher drag reductions, of the order of 40-50%, the ow enters the HDR regime for which the slope of the log-law is dramatically augmented and the Reynolds shear stress is small (Warholic et al. 1999; Ptasinski et al. 2001). The drag reduction is eventually bounded by a maximum drag reduction (MDR) (Virk & Mickley 1970) which is a function of the Reynolds number. While several experiments report mean velocity profiles very close to the empirical profile of Virk & Mickley (1970) for MDR conditions, the observations regarding the structure of turbulence can differ significantly. For instance, Warholic et al. (1999) measured a near-zero Reynolds shear stress, whereas a recent experiment (Ptasinski et al. 2001) shows evidence of non-negligible Reynolds stress in their MDR flow. To the knowledge of the authors, only the LDR regime has been documented in numerical simulations (Sureshkumar et al. 1997; Dimitropoulos et al. 1998; Min et al. 2001; Dubief & Lele 2001; Sibilla & Baron 2002). This paper discusses the simulation of polymer drag reduced channel ow at HDR using the FENE-P (Finite Elastic non

  3. High-throughput titration of luciferase-expressing recombinant viruses.

    PubMed

    Garcia, Vanessa; Krishnan, Ramya; Davis, Colin; Batenchuk, Cory; Le Boeuf, Fabrice; Abdelbary, Hesham; Diallo, Jean-Simon

    2014-01-01

    Standard plaque assays to determine infectious viral titers can be time consuming, are not amenable to a high volume of samples, and cannot be done with viruses that do not form plaques. As an alternative to plaque assays, we have developed a high-throughput titration method that allows for the simultaneous titration of a high volume of samples in a single day. This approach involves infection of the samples with a Firefly luciferase tagged virus, transfer of the infected samples onto an appropriate permissive cell line, subsequent addition of luciferin, reading of plates in order to obtain luminescence readings, and finally the conversion from luminescence to viral titers. The assessment of cytotoxicity using a metabolic viability dye can be easily incorporated in the workflow in parallel and provide valuable information in the context of a drug screen. This technique provides a reliable, high-throughput method to determine viral titers as an alternative to a standard plaque assay.

  4. Forager bees (Apis mellifera) highly express immune and detoxification genes in tissues associated with nectar processing.

    PubMed

    Vannette, Rachel L; Mohamed, Abbas; Johnson, Brian R

    2015-11-09

    Pollinators, including honey bees, routinely encounter potentially harmful microorganisms and phytochemicals during foraging. However, the mechanisms by which honey bees manage these potential threats are poorly understood. In this study, we examine the expression of antimicrobial, immune and detoxification genes in Apis mellifera and compare between forager and nurse bees using tissue-specific RNA-seq and qPCR. Our analysis revealed extensive tissue-specific expression of antimicrobial, immune signaling, and detoxification genes. Variation in gene expression between worker stages was pronounced in the mandibular and hypopharyngeal gland (HPG), where foragers were enriched in transcripts that encode antimicrobial peptides (AMPs) and immune response. Additionally, forager HPGs and mandibular glands were enriched in transcripts encoding detoxification enzymes, including some associated with xenobiotic metabolism. Using qPCR on an independent dataset, we verified differential expression of three AMP and three P450 genes between foragers and nurses. High expression of AMP genes in nectar-processing tissues suggests that these peptides may contribute to antimicrobial properties of honey or to honey bee defense against environmentally-acquired microorganisms. Together, these results suggest that worker role and tissue-specific expression of AMPs, and immune and detoxification enzymes may contribute to defense against microorganisms and xenobiotic compounds acquired while foraging.

  5. Forager bees (Apis mellifera) highly express immune and detoxification genes in tissues associated with nectar processing

    PubMed Central

    Vannette, Rachel L.; Mohamed, Abbas; Johnson, Brian R.

    2015-01-01

    Pollinators, including honey bees, routinely encounter potentially harmful microorganisms and phytochemicals during foraging. However, the mechanisms by which honey bees manage these potential threats are poorly understood. In this study, we examine the expression of antimicrobial, immune and detoxification genes in Apis mellifera and compare between forager and nurse bees using tissue-specific RNA-seq and qPCR. Our analysis revealed extensive tissue-specific expression of antimicrobial, immune signaling, and detoxification genes. Variation in gene expression between worker stages was pronounced in the mandibular and hypopharyngeal gland (HPG), where foragers were enriched in transcripts that encode antimicrobial peptides (AMPs) and immune response. Additionally, forager HPGs and mandibular glands were enriched in transcripts encoding detoxification enzymes, including some associated with xenobiotic metabolism. Using qPCR on an independent dataset, we verified differential expression of three AMP and three P450 genes between foragers and nurses. High expression of AMP genes in nectar-processing tissues suggests that these peptides may contribute to antimicrobial properties of honey or to honey bee defense against environmentally-acquired microorganisms. Together, these results suggest that worker role and tissue-specific expression of AMPs, and immune and detoxification enzymes may contribute to defense against microorganisms and xenobiotic compounds acquired while foraging. PMID:26549293

  6. Statistically invalid classification of high throughput gene expression data.

    PubMed

    Barbash, Shahar; Soreq, Hermona

    2013-01-01

    Classification analysis based on high throughput data is a common feature in neuroscience and other fields of science, with a rapidly increasing impact on both basic biology and disease-related studies. The outcome of such classifications often serves to delineate novel biochemical mechanisms in health and disease states, identify new targets for therapeutic interference, and develop innovative diagnostic approaches. Given the importance of this type of studies, we screened 111 recently-published high-impact manuscripts involving classification analysis of gene expression, and found that 58 of them (53%) based their conclusions on a statistically invalid method which can lead to bias in a statistical sense (lower true classification accuracy then the reported classification accuracy). In this report we characterize the potential methodological error and its scope, investigate how it is influenced by different experimental parameters, and describe statistically valid methods for avoiding such classification mistakes.

  7. Different gene expressions between cattle and yak provide insights into high-altitude adaptation.

    PubMed

    Wang, K; Yang, Y; Wang, L; Ma, T; Shang, H; Ding, L; Han, J; Qiu, Q

    2016-02-01

    DNA sequence variation has been widely reported as the genetic basis for adaptation, in both humans and other animals, to the hypoxic environment experienced at high altitudes. However, little is known about the patterns of gene expression underlying such hypoxic adaptations. In this study, we examined the differences in the transcriptomes of four organs (heart, kidney, liver and lung) between yak and cattle, a pair of closely related species distributed at high and low altitudes respectively. Of the four organs examined, heart shows the greatest differentiation between the two species in terms of gene expression profiles. Detailed analyses demonstrated that some genes associated with the oxygen supply system and the defense systems that respond to threats of hypoxia are differentially expressed. In addition, genes with significantly differentiated patterns of expression in all organs exhibited an unexpected uniformity of regulation along with an elevated frequency of nonsynonymous substitutions. This co-evolution of protein sequences and gene expression patterns is likely to be correlated with the optimization of the yak metabolic system to resist hypoxia.

  8. Identification of Gene Expression Signatures in the Chicken Intestinal Intraepithelial Lymphocytes in Response to Herb Additive Supplementations

    PubMed Central

    Won, Kyeong-Hye; Song, Ki-Duk; Park, Jong-Eun; Kim, Duk-Kyung; Na, Chong-Sam

    2016-01-01

    Anethole and garlic have an immune modulatory effects on avian coccidiosis, and these effects are correlated with gene expression changes in intestinal epithelial lymphocytes (IELs). In this study, we integrated gene expression datasets from two independent experiments and investigated gene expression profile changes by anethole and garlic respectively, and identified gene expression signatures, which are common targets of these herbs as they might be used for the evaluation of the effect of plant herbs on immunity toward avian coccidiosis. We identified 4,382 and 371 genes, which were differentially expressed in IELs of chickens supplemented with garlic and anethole respectively. The gene ontology (GO) term of differentially expressed genes (DEGs) from garlic treatment resulted in the biological processes (BPs) related to proteolysis, e.g., “modification-dependent protein catabolic process”, “proteolysis involved in cellular protein catabolic process”, “cellular protein catabolic process”, “protein catabolic process”, and “ubiquitin-dependent protein catabolic process”. In GO analysis, one BP term, “Proteolysis”, was obtained. Among DEGs, 300 genes were differentially regulated in response to both garlic and anethole, and 234 and 59 genes were either up- or down-regulated in supplementation with both herbs. Pathway analysis resulted in enrichment of the pathways related to digestion such as “Starch and sucrose metabolism” and “Insulin signaling pathway”. Taken together, the results obtained in the present study could contribute to the effective development of evaluation system of plant herbs based on molecular signatures related with their immunological functions in chicken IELs. PMID:26954117

  9. An inducible expression system for high-level expression of recombinant proteins in slow growing mycobacteria.

    PubMed

    Leotta, Lisa; Spratt, Joanne M; Kong, Carlyn U; Triccas, James A

    2015-09-01

    A novel protein expression vector utilising the inducible hspX promoter of Mycobacterium tuberculosis was constructed and evaluated in this study. High-level induction of three mycobacterial antigens, comprising up to 9% of bacterial sonicate, was demonstrated in recombinant Mycobacterium bovis BCG when grown under low-oxygen tension, which serves to enhance hspX promoter activity. Recombinant proteins were efficiently purified from bacterial lysates in a soluble form by virtue of a C-terminal 6-histidine tag. Purification of the immunodominant M. tuberculosis Ag85B antigen using this system resulted in a recombinant protein that stimulated significant IFN-γ release from Ag85B-reactive T cells generated after vaccination of mice with an Ag85B-expressing vaccine. Further, the M. tuberculosis L-alanine dehydrogenase (Ald) protein purified from recombinant BCG displayed strong enzymatic activity in recombinant form. This study demonstrated that high levels of native-like recombinant mycobacterial proteins can be produced in mycobacterial hosts, and this may aid the analysis of mycobacterial protein function and the development of new treatments. PMID:26021569

  10. High-throughput differential screening of mRNAs by serial analysis of gene expression: decreased expression of trefoil factor 3 mRNA in thyroid follicular carcinomas.

    PubMed

    Takano, T; Miyauchi, A; Yoshida, H; Kuma, K; Amino, N

    2004-04-19

    To find mRNAs whose expression differs between thyroid follicular adenomas and carcinomas, a high-throughput analysis of mRNAs in these two tumours was performed. This method, named high-throughput differential screening by serial analysis of gene expression (HDSS), combines a modified method of serial analysis of gene expression (SAGE) and real-time quantitative reverse transcription polymerase chain reaction (RT-PCR). A total of 40 candidate tag sequences that showed extremely different expression levels between a follicular carcinoma and a follicular adenoma in the SAGE analysis were analysed by real-time quantitative RT-PCR, using RNAs from an additional four typical follicular carcinomas and adenomas. One sequence tag that represents trefoil factor 3 (TFF3) mRNA showed a clear difference in its expression level between adenomas and carcinomas. The expression levels of TFF3 mRNA in 48 follicular adenomas and 29 follicular carcinomas were measured by real-time quantitative RT-PCR using a specific probe for TFF3. They were significantly decreased in follicular carcinomas, especially in widely invasive types and those with evident metastases. These results indicate that the decreased expression of TFF3 mRNA is a marker of follicular carcinomas, especially those with a high risk of invasion or metastasis.

  11. Gene expression suggests conserved aspects of Hox gene regulation in arthropods and provides additional support for monophyletic Myriapoda.

    PubMed

    Janssen, Ralf; Budd, Graham E

    2010-01-01

    Antisense transcripts of Ultrabithorax (aUbx) in the millipede Glomeris and the centipede Lithobius are expressed in patterns complementary to that of the Ubx sense transcripts. A similar complementary expression pattern has been described for non-coding RNAs (ncRNAs) of the bithoraxoid (bxd) locus in Drosophila, in which the transcription of bxd ncRNAs represses Ubx via transcriptional interference. We discuss our findings in the context of possibly conserved mechanisms of Ubx regulation in myriapods and the fly.Bicistronic transcription of Ubx and Antennapedia (Antp) has been reported previously for a myriapod and a number of crustaceans. In this paper, we show that Ubx/Antp bicistronic transcripts also occur in Glomeris and an onychophoran, suggesting further conserved mechanisms of Hox gene regulation in arthropods.Myriapod monophyly is supported by the expression of aUbx in all investigated myriapods, whereas in other arthropod classes, including the Onychophora, aUbx is not expressed. Of the two splice variants of Ubx/Antp only one could be isolated from myriapods, representing a possible further synapomorphy of the Myriapoda. PMID:20849647

  12. Acetate Salts as Nonhalogen Additives To Improve Perovskite Film Morphology for High-Efficiency Solar Cells.

    PubMed

    Wu, Qiliang; Zhou, Pengcheng; Zhou, Weiran; Wei, Xiangfeng; Chen, Tao; Yang, Shangfeng

    2016-06-22

    A two-step method has been popularly adopted to fabricate a perovskite film of planar heterojunction organo-lead halide perovskite solar cells (PSCs). However, this method often generates uncontrollable film morphology with poor coverage. Herein, we report a facile method to improve perovskite film morphology by incorporating a small amount of acetate (CH3COO(-), Ac(-)) salts (NH4Ac, NaAc) as nonhalogen additives in CH3NH3I solution used for immersing PbI2 film, resulting in improved CH3NH3PbI3 film morphology. Under the optimized NH4Ac additive concentration of 10 wt %, the best power conversion efficiency (PCE) reaches 17.02%, which is enhanced by ∼23.2% relative to that of the pristine device without additive, whereas the NaAc additive does not lead to an efficiency enhancement despite the improvement of the CH3NH3PbI3 film morphology. SEM study reveals that NH4Ac and NaAc additives can both effectively improve perovskite film morphology by increasing the surface coverage via diminishing pinholes. The improvement on CH3NH3PbI3 film morphology is beneficial for increasing the optical absorption of perovskite film and improving the interfacial contact at the perovskite/spiro-OMeTAD interface, leading to the increase of short-circuit current and consequently efficiency enhancement of the PSC device for NH4Ac additive only.

  13. Modulation of gene expression in endothelial cells in response to high LET nickel ion irradiation.

    PubMed

    Beck, Michaël; Rombouts, Charlotte; Moreels, Marjan; Aerts, An; Quintens, Roel; Tabury, Kevin; Michaux, Arlette; Janssen, Ann; Neefs, Mieke; Ernst, Eric; Dieriks, Birger; Lee, Ryonfa; De Vos, Winnok H; Lambert, Charles; Van Oostveldt, Patrick; Baatout, Sarah

    2014-10-01

    Ionizing radiation can elicit harmful effects on the cardiovascular system at high doses. Endothelial cells are critical targets in radiation-induced cardiovascular damage. Astronauts performing a long-term deep space mission are exposed to consistently higher fluences of ionizing radiation that may accumulate to reach high effective doses. In addition, cosmic radiation contains high linear energy transfer (LET) radiation that is known to produce high values of relative biological effectiveness (RBE). The aim of this study was to broaden the understanding of the molecular response to high LET radiation by investigating the changes in gene expression in endothelial cells. For this purpose, a human endothelial cell line (EA.hy926) was irradiated with accelerated nickel ions (Ni) (LET, 183 keV/µm) at doses of 0.5, 2 and 5 Gy. DNA damage was measured 2 and 24 h following irradiation by γ-H2AX foci detection by fluorescence microscopy and gene expression changes were measured by microarrays at 8 and 24 h following irradiation. We found that exposure to accelerated nickel particles induced a persistent DNA damage response up to 24 h after treatment. This was accompanied by a downregulation in the expression of a multitude of genes involved in the regulation of the cell cycle and an upregulation in the expression of genes involved in cell cycle checkpoints. In addition, genes involved in DNA damage response, oxidative stress, apoptosis and cell-cell signaling (cytokines) were found to be upregulated. An in silico analysis of the involved genes suggested that the transcription factors, E2F and nuclear factor (NF)-κB, may be involved in these cellular responses. PMID:25118949

  14. FX cloning: a simple and robust high-throughput cloning method for protein expression.

    PubMed

    Geertsma, Eric R

    2014-01-01

    The immense amount of gene sequences available nowadays allows scientist to screen broadly for extraordinary proteins. Reliable cloning tools that allow the parallel processing of many targets are vital for the success of this strategy. The FX cloning procedure detailed here is such a straightforward and efficient tool. It is dedicated to the cloning of open reading frames (ORFs) with the final aim of expressing the corresponding proteins. FX cloning combines attractive features of established high-throughput cloning methods that were thus far not unified in one single method. It facilitates the subcloning of a sequence-verified ORF to a variety of expression vectors, but is sufficiently versatile to accept PCR products as well. Moreover, the common, but undesirable feature of extending target ORFs with long cloning-related sequences is avoided. It leads to the addition of only one amino acid to each side of the protein. As a consequence, only one primer pair or PCR product suffices to generate expression vectors for both N- and C-terminal translational fusions. FX cloning is highly efficient and economical in its use. The method is suited for high-throughput cloning projects and also for everyday cloning of single targets. FX cloning is based on the use of type IIS restriction enzymes and negative selection markers. The full procedure takes place in one pot in less than 3 h and does not require intermediate purification steps nor extensive handling. The method has proven to be very robust and suitable for all common expression systems.

  15. CIP2A protein expression in high-grade, high-stage bladder cancer

    PubMed Central

    Huang, Lisa P; Savoly, Diana; Sidi, Abraham A; Adelson, Martin E; Mordechai, Eli; Trama, Jason P

    2012-01-01

    Bladder cancer is one of the most common cancers in the United States. Numerous markers have been evaluated for suitability of bladder cancer detection and surveillance. However, few of them are acceptable as a routine tool. Therefore, there exists a continuing need for an assay that detects the presence of bladder cancer in humans. It would be advantageous to develop an assay with a protein that is associated with the development of bladder cancer. We have identified the cancerous inhibitor of PP2A (CIP2A) protein as a novel bladder cancer biomarker. In this study, Western blot analysis was used to assess the expression level of CIP2A protein in bladder cancer cell lines and bladder cancer patient tissues (n = 43). Our studies indicated CIP2A protein was abundantly expressed in bladder cancer cell lines but not in nontumor epithelial cell lines. Furthermore, CIP2A was specifically expressed in transitional cell carcinoma (TCC) of the bladder tumor tissues but not in adjacent nontumor bladder tissue. Our data showed that CIP2A protein detection in high-grade TCC tissues had a sensitivity of 65%, which is 3.4-fold higher than that seen in low-grade TCC tissues (19%). The level of CIP2A protein expression increased with the stage of disease (12%, 27%, 67%, and 100% for pTa, pT1, pT2, and pT3 tumor, respectively). In conclusion, our studies suggest that CIP2A protein is specifically expressed in human bladder tumors. CIP2A is preferentially expressed in high-grade and high-stage TCC tumors, which are high-risk and invasive tumors. Our studies reported here support the role of CIP2A in bladder cancer progression and its usefulness for the surveillance of recurrence or progression of human bladder cancer. PMID:23342256

  16. Dopamine neurons in culture express VGLUT2 explaining their capacity to release glutamate at synapses in addition to dopamine.

    PubMed

    Dal Bo, Gregory; St-Gelais, Fannie; Danik, Marc; Williams, Sylvain; Cotton, Mathieu; Trudeau, Louis-Eric

    2004-03-01

    Dopamine neurons have been suggested to use glutamate as a cotransmitter. To identify the basis of such a phenotype, we have examined the expression of the three recently identified vesicular glutamate transporters (VGLUT1-3) in postnatal rat dopamine neurons in culture. We found that the majority of isolated dopamine neurons express VGLUT2, but not VGLUT1 or 3. In comparison, serotonin neurons express only VGLUT3. Single-cell RT-PCR experiments confirmed the presence of VGLUT2 mRNA in dopamine neurons. Arguing for phenotypic heterogeneity among axon terminals, we find that only a proportion of terminals established by dopamine neurons are VGLUT2-positive. Taken together, our results provide a basis for the ability of dopamine neurons to release glutamate as a cotransmitter. A detailed analysis of the conditions under which DA neurons gain or loose a glutamatergic phenotype may provide novel insight into pathophysiological processes that underlie diseases such as schizophrenia, Parkinson's disease and drug dependence. PMID:15009640

  17. High pressure-sensitive gene expression in Lactobacillus sanfranciscensis.

    PubMed

    Vogel, R F; Pavlovic, M; Hörmann, S; Ehrmann, M A

    2005-08-01

    Lactobacillus sanfranciscensis is a Gram-positive lactic acid bacterium used in food biotechnology. It is necessary to investigate many aspects of a model organism to elucidate mechanisms of stress response, to facilitate preparation, application and performance in food fermentation, to understand mechanisms of inactivation, and to identify novel tools for high pressure biotechnology. To investigate the mechanisms of the complex bacterial response to high pressure we have analyzed changes in the proteome and transcriptome by 2-D electrophoresis, and by microarrays and real time PCR, respectively. More than 16 proteins were found to be differentially expressed upon high pressure stress and were compared to those sensitive to other stresses. Except for one apparently high pressure-specific stress protein, no pressure-specific stress proteins were found, and the proteome response to pressure was found to differ from that induced by other stresses. Selected pressure-sensitive proteins were partially sequenced and their genes were identified by reverse genetics. In a transcriptome analysis of a redundancy cleared shot gun library, about 7% of the genes investigated were found to be affected. Most of them appeared to be up-regulated 2- to 4-fold and these results were confirmed by real time PCR. Gene induction was shown for some genes up-regulated at the proteome level (clpL/groEL/rbsK), while the response of others to high hydrostatic pressure at the transcriptome level seemed to differ from that observed at the proteome level. The up-regulation of selected genes supports the view that the cell tries to compensate for pressure-induced impairment of translation and membrane transport. PMID:16082466

  18. High expression of TRPM8 predicts poor prognosis in patients with osteosarcoma

    PubMed Central

    Zhao, Wei; Xu, Huimian

    2016-01-01

    The transient receptor potential melastatin member 8 (TRPM8) is a newly characterized oncoprotein involved in various malignant tumors. However, its expression pattern and biological function in osteosarcoma remain unclear. The present study aimed to explore the expression and prognostic significance of TRPM8 in osteosarcoma (OS). The results revealed that the expression of TRPM8 mRNA and protein in OS tissue was significantly higher than that in paired normal bone tissue (P<0.05). Additionally, the level of TRPM8 mRNA and protein in patients with a higher clinical stage and with distant metastasis was markedly higher than in those with a lower clinical stage and no metastasis (P<0.05). Furthermore, a high TRPM8 level was closely associated with clinical stage and distant metastasis (P=0.007 and 0.030), but not associated with the patient age (P=0.481), gender (P=0.905), tumor size (P=0.429), histological subtype (P=0.207) or anatomical location (P=0.369). In addition, OS patients with high TRPM8 expression had significantly shorter overall survival (P=0.008) and disease-free survival times (P=0.008) when compared with patients with low expression of TRPM8. In Cox multivariate analysis, TRPM8 overexpression was identified to be an independent and significant prognostic factor for overall survival (P=0.040), but not for disease-free survival (P=0.051). Collectively, the present data suggest that TRPM8 may play a crucial role in the development and progression of OS, and thus may be considered as a novel molecular target for therapy in patients with OS. PMID:27446440

  19. High lysophosphatidylcholine acyltransferase 1 expression independently predicts high risk for biochemical recurrence in prostate cancers.

    PubMed

    Grupp, Katharina; Sanader, Stella; Sirma, Hüseyin; Simon, Ronald; Koop, Christina; Prien, Kristina; Hube-Magg, Claudia; Salomon, Georg; Graefen, Markus; Heinzer, Hans; Minner, Sarah; Izbicki, Jakob R; Sauter, Guido; Schlomm, Thorsten; Tsourlakis, Maria Christina

    2013-12-01

    Lysophosphatidylcholine acyltransferase 1 (LPCAT1) has been suggested to play a role in cancer. To assess its role in prostate cancer, LPCAT1 expression was analyzed on a tissue microarray containing samples from 11,152 prostate cancer patients. In benign prostate glands, LPCAT1 immunostaining was absent or weak. In prostate cancer, LPCAT1 positivity was found in 73.8% of 8786 interpretable tumors including 29.2% with strong expression. Increased LPCAT1 expression was associated with advanced tumor stage (pT3b/T4) (p < 0.0001), high Gleason score (≥4 + 4) (p < 0.0001), positive nodal involvement (p = 0.0002), positive surgical margin (p = 0.0005), and early PSA recurrence (p < 0.0001). High LPCAT1 expression was strongly linked to ERG-fusion type prostate cancer. Strong LPCAT1 staining was detected in 45.3% of ERG positive but in only 16.7% of ERG negative tumors (p < 0.0001). Within ERG negative cancers, LPCAT1 staining was strongly increased within the subgroup of PTEN deleted cancers (p < 0.0001). Further subgroup analyses revealed that associations of high LPCAT1 expression with PSA recurrence and unfavorable tumor phenotype were largely driven by ERG negative cancers (p < 0.0001) while these effects were substantially mitigated in ERG positive cancers (p = 0.0073). The prognostic impact of LPCAT1 expression was independent of histological and clinical parameters. It is concluded, that LPCAT1 measurement, either alone or in combination, may be utilized for better clinical decision-making. These data also highlight the potentially important role of lipid metabolism in prostate cancer biology.

  20. High level PHGDH expression in breast is predominantly associated with keratin 5-positive cell lineage independently of malignancy.

    PubMed

    Gromova, Irina; Gromov, Pavel; Honma, Naoko; Kumar, Sudha; Rimm, David; Talman, Maj-Lis Møller; Wielenga, Vera Timmermans; Moreira, José M A

    2015-10-01

    We have previously reported the 2D PAGE-based proteomic profiling of a prospective cohort of 78 triple negative breast cancer (TNBC) patients, and the establishment of a cumulative TNBC protein database. Analysis of this database identified a number of proteins as being specifically overexpressed in TNBC samples. One such protein was D-3-phosphoglycerate dehydrogenase (Phgdh), a candidate oncogene. We analysed expression of Phgdh in normal and TNBC mammary tissue samples by 2D gel-based proteomics and immunohistochemistry (IHC), and show here that high-level expression of Phgdh in mammary epithelial cells is primarily associated with cell lineage, as we found that Phgdh expression was predominant in CK5-positive cells, normal as well as malignant, thus identifying an association of this protein with the basal phenotype. Quantitative IHC analysis of Phgdh expression in normal breast tissue showed high-level expression of Phgdh in normal CK5-positive mammary epithelial cells, indicating that expression of this protein was not associated with malignancy, but rather with cell lineage. However, proteomic profiling of Phgdh showed it to be expressed in two major protein forms, and that the ratio of expression between these variants was associated with malignancy. Overexpression of Phgdh in CK5-positive cell lineages, and differential protein isoform expression, was additionally found in other tissues and cancer types, suggesting that overexpression of Phgdh is generally associated with CK5 cells, and that oncogenic function may be determined by isoform expression.

  1. Enhancement of anammox activity by addition of compatible solutes at high salinity conditions.

    PubMed

    Liu, Mu; Peng, Yongzhen; Wang, Shuying; Liu, Tiantian; Xiao, Han

    2014-09-01

    The enhancement effect of compatible solutes on anammox activity under salinity stress was investigated. Glycine betaine (GB) was the most effective in alleviating salt toxicity, although all the compatible solutes (GB, trehalose and ectoine) were found to be valid. Acclimation potential of anammox biomass under salinity of 30 g/L increased significantly with GB addition. The recovery time in the reactor with GB addition (RB) (49 days) accompanied by a more stable stoichiometric ratio was 2.65 times shorter than in the control reactor (RC) (130 days). After 49 days, the extracellular polymeric substances and the tetrazolium chloride-dehydrogenase activity were 217.9 mg/g VSS and 38.7 μg TF/g VSS/h in RB, 1.86 times lower and 3.17 times higher than the levels in RC, respectively. RB possessed evident superiority in the aspects of microbial population proportion. And thus, compatible solutes addition was regarded as one of the feasible solution to counteract saline inhibition on anammox. PMID:25024098

  2. Big Area Additive Manufacturing of High Performance Bonded NdFeB Magnets

    PubMed Central

    Li, Ling; Tirado, Angelica; Nlebedim, I. C.; Rios, Orlando; Post, Brian; Kunc, Vlastimil; Lowden, R. R.; Lara-Curzio, Edgar; Fredette, Robert; Ormerod, John; Lograsso, Thomas A.; Paranthaman, M. Parans

    2016-01-01

    Additive manufacturing allows for the production of complex parts with minimum material waste, offering an effective technique for fabricating permanent magnets which frequently involve critical rare earth elements. In this report, we demonstrate a novel method - Big Area Additive Manufacturing (BAAM) - to fabricate isotropic near-net-shape NdFeB bonded magnets with magnetic and mechanical properties comparable or better than those of traditional injection molded magnets. The starting polymer magnet composite pellets consist of 65 vol% isotropic NdFeB powder and 35 vol% polyamide (Nylon-12). The density of the final BAAM magnet product reached 4.8 g/cm3, and the room temperature magnetic properties are: intrinsic coercivity Hci = 688.4 kA/m, remanence Br = 0.51 T, and energy product (BH)max = 43.49 kJ/m3 (5.47 MGOe). In addition, tensile tests performed on four dog-bone shaped specimens yielded an average ultimate tensile strength of 6.60 MPa and an average failure strain of 4.18%. Scanning electron microscopy images of the fracture surfaces indicate that the failure is primarily related to the debonding of the magnetic particles from the polymer binder. The present method significantly simplifies manufacturing of near-net-shape bonded magnets, enables efficient use of rare earth elements thus contributing towards enriching the supply of critical materials. PMID:27796339

  3. How to produce personality neuroscience research with high statistical power and low additional cost.

    PubMed

    Mar, Raymond A; Spreng, R Nathan; Deyoung, Colin G

    2013-09-01

    Personality neuroscience involves examining relations between cognitive or behavioral variability and neural variables like brain structure and function. Such studies have uncovered a number of fascinating associations but require large samples, which are expensive to collect. Here, we propose a system that capitalizes on neuroimaging data commonly collected for separate purposes and combines it with new behavioral data to test novel hypotheses. Specifically, we suggest that groups of researchers compile a database of structural (i.e., anatomical) and resting-state functional scans produced for other task-based investigations and pair these data with contact information for the participants who contributed the data. This contact information can then be used to collect additional cognitive, behavioral, or individual-difference data that are then reassociated with the neuroimaging data for analysis. This would allow for novel hypotheses regarding brain-behavior relations to be tested on the basis of large sample sizes (with adequate statistical power) for low additional cost. This idea can be implemented at small scales at single institutions, among a group of collaborating researchers, or perhaps even within a single lab. It can also be implemented at a large scale across institutions, although doing so would entail a number of additional complications.

  4. Combination Therapy with Losartan and Pioglitazone Additively Reduces Renal Oxidative and Nitrative Stress Induced by Chronic High Fat, Sucrose, and Sodium Intake

    PubMed Central

    Kong, Xiang; Zhang, Yan; Wu, Hai-bing; Li, Fang-xia; Zhang, Dao-you; Su, Qing

    2012-01-01

    We recently showed that combination therapy with losartan and pioglitazone provided synergistic effects compared with monotherapy in improving lesions of renal structure and function in Sprague-Dawley rats fed with a high-fat, high-sodium diet and 20% sucrose solution. This study was designed to explore the underlying mechanisms of additive renoprotection provided by combination therapy. Losartan, pioglitazone, and their combination were orally administered for 8 weeks. The increased level of renal malondialdehyde and expression of nicotinamide adenine dinucleotide phosphate oxidase subunit p47phox and nitrotyrosine as well as the decreased total superoxide dismutase activity and copper, zinc-superoxide dismutase expression were tangible evidence for the presence of oxidative and nitrative stress in the kidney of model rats. Treatment with both drugs, individually and in combination, improved these abnormal changes. Combination therapy showed synergistic effects in reducing malondialdehyde level, p47phox, and nitrotyrosine expression to almost the normal level compared with monotherapy. All these results suggest that the additive renoprotection provided by combination therapy might be attributed to a further reduction of oxidative and nitrative stress. PMID:23213350

  5. Expression of P-glycoprotein in high grade osteosarcomas with special emphasis on chondroblastic subtype.

    PubMed

    Radig, K; Häckel, C; Herting, J; Oda, Y; Mittler, U; Neumann, W; Roessner, A

    1997-02-01

    The development of chemoresistance is one of the major clinical problems in the therapy of malignant bone tumors in childhood. The expression of membrane-bound P-glycoprotein turned out to be an essential factor in the evidence of resistant tumor cells. To investigate the significance of multidrug resistance in the prognosis of highly malignant osteosarcomas, the immunohistologic expression of P-glycoprotein was investigated in the tumor tissue of 52 patients under special consideration of the histologic subtype. The data were compared with the histologic regression grade in the resection specimen and correlated with clinical data. Formalin-fixed, paraffin-embedded tissue and, additionally, fresh frozen material taken from the primary biopsy were stained using monoclonal antibody JSB1. 29 (55%) of the tumors investigated were P-glycoprotein positive. Considering the response to chemotherapy, no conclusion could be drawn regarding P-glycoprotein expression, regression grade in the resection specimens, and the clinical follow-up. P-glycoprotein was detected in only 52% of the non-responders. A positive reaction was also evidenced in 59% of the patients with high chemosensitivity. A comparison of the histologic subtypes yielded a significant result in the chondroblastic osteosarcomas. 11 of 12 cases showed a strong expression of P-glycoprotein. Most of the cases were non-responders, and using Kaplan-Meier live tables, an unfavorable clinical outcome could be demonstrated. Possibly, chondroblastic tumors have a special position among osteosarcomas because of their differentiation. PMID:9065577

  6. High RAD54B expression: an independent predictor of postoperative distant recurrence in colorectal cancer patients

    PubMed Central

    Nagai, Yuzo; Yamamoto, Yoko; Yasuhara, Takaaki; Hata, Keisuke; Nishikawa, Takeshi; Tanaka, Toshiaki; Tanaka, Junichiro; Kiyomatsu, Tomomichi; Kawai, Kazushige; Nozawa, Hiroaki; Kazama, Shinsuke; Yamaguchi, Hironori; Ishihara, Soichiro; Sunami, Eiji; Yamanaka, Takeharu; Miyagawa, Kiyoshi; Watanabe, Toshiaki

    2015-01-01

    We recently reported a specific mechanism that RAD54B, an important factor in homologous recombination, promotes genomic instability via the degradation of p53 protein in vitro. However, clinical significance of RAD54Bin colorectal cancer (CRC) remains unclear. Thus we analyzed RAD54B geneexpression in CRC patients. Using the training set (n = 123), the optimal cut-off value for stratification was determined, and validated in another cohort (n = 89). Kaplan–Meier plots showed that distant recurrence free survival was significantly lesser in high RAD54B expression group compared with that of low expression group in both training (P = 0.0013) and validation (P = 0.024) set. Multivariate analysis using Cox proportional-hazards model showed that high RAD54B expression was an independent predictor in both training (hazard ratio, 4.31; 95% CI, 1.53–13.1; P = 0.0060) and validation (hazard ratio, 3.63; 95% CI, 1.23–10.7; P = 0.021) set. In addition, a negative significant correlation between RAD54B and CDKN1A, a target gene of p53, was partially confirmed, suggesting that RAD54B functions via the degradation of p53 protein even in clinical samples. This study first demonstrated RAD54B expression has potential to serve as a novel prognostic biomarker, particularly for distant recurrence in CRC patients. PMID:26046797

  7. High expression of epithelial cellular adhesion molecule in peritoneal metastasis of gastric cancer.

    PubMed

    Imano, Motohiro; Itoh, Tatsuki; Satou, Takao; Yasuda, Atsushi; Nishiki, Kohei; Kato, Hiroaki; Shiraishi, Osamu; Peng, Ying-Feng; Shinkai, Masayuki; Tsubaki, Masahiro; Yasuda, Takushi; Imamoto, Haruhiko; Nishida, Shozo; Takeyama, Yoshifumi; Furkawa, Hiroshi; Okuno, Kiyokata; Shiozaki, Hitoshi

    2013-12-01

    Intraperitoneally administrated epithelial cellular adhesion molecule (EpCAM) monoclonal antibody is a therapeutic agent in patients with malignant effusion in several types of carcinoma. However, the role of EpCAM in peritoneal metastasis (PM) lesions and primary lesions of gastric cancer (GC) is still unclear. Therefore, in this study, we investigated EpCAM expression in GC patients with PM. We investigated the expression of EpCAM in 35PM lesions and 104 biopsy samples as primary lesions. Immunohistochemical staining was performed using the Ventana Benchmark XT (Roche Diagnostics) system. EpCAM expression was evaluated by calculating the total immunostaining score, which is the product of the proportion score and the intensity score. Overexpression was defined as a total score greater than 4. All PM specimens showed overexpression of EpCAM, and GC cells in both the surface layer and the deep layer of the PM showed a high expression of EpCAM. Meanwhile, in the biopsy sample, the expression of EpCAM ranged from none to strong. The EpCAM score results for PM specimens and biopsy samples were 11.0 ± 2.0 and 6.9 ± 3.9, respectively. The difference between the scores was statistically significant (P < 0.05). The intraperitoneally administrated EpCAM antibody might have a anti-cancer effect in PM lesions of GC. Additionally, it can be assumed that only GC cells which express a high level of EpCAM might metastasize to the peritoneum.

  8. The addition of accessory enzymes enhances the hydrolytic performance of cellulase enzymes at high solid loadings.

    PubMed

    Hu, Jinguang; Chandra, Richard; Arantes, Valdeir; Gourlay, Keith; van Dyk, J Susan; Saddler, Jack N

    2015-06-01

    The pretreatment process used and the nature of the biomass feedstock will influence the role that accessory enzymes can play in synergistically interacting with cellulases to effectively deconstruct the substrate. The work reported here assessed the possible boosting effects of the xylanase and lytic polysaccharide monooxygenase (AA9, formerly known as GH61) on the hydrolytic potential of cellulase enzyme mixtures during hydrolysis of steam pretreated poplar and corn stover at high (10-20% w/v) substrate concentrations. A higher proportion of xylanase was required when the substrate had a relatively high xylan content and at high substrate concentrations. In contrast, a relatively small amount of AA9 (about 2 mg/g cellulose) was enough, regardless of the nature or concentration of the substrate. The overall protein loading required to achieve effective hydrolysis of high concentrations of pretreated biomass substrates could be substantially reduced by optimizing the ratio of enzymes in the "cellulase" mixture.

  9. Synergistic catalysis: highly diastereoselective benzoxazole addition to Morita-Baylis-Hillman carbonates.

    PubMed

    Ceban, Victor; Putaj, Piotr; Meazza, Marta; Pitak, Mateusz B; Coles, Simon J; Vesely, Jan; Rios, Ramon

    2014-07-18

    An expedited method has been developed for the diastereoselective synthesis of highly functionalized alkyl-azaarene systems with good yields and high diastereoselectivities (>15 : 1 dr). The methodology includes a synergistic catalysis event involving organometallic (10 mol% AgOAc) activation of an alkyl azaarene and Lewis base (10 mol% DABCO) activation of a Morita-Baylis-Hillman carbonate. The structure and relative configuration of a representative product were confirmed by X-ray analysis.

  10. Michael Additions of Highly Basic Enolates to ortho-Quinone Methides

    PubMed Central

    Lewis, Robert S.; Garza, Christopher J.; Dang, Ann T.; Pedro, Te Kie A.; Chain, William J.

    2015-01-01

    A protocol by which ketone or ester enolates and ortho-quinone methides (o-QMs) are generated in situ in a single reaction flask from silylated precursors under the action of anhydrous fluoride is reported. The reaction partners are joined to give a variety of β-(2-hydroxyphenyl)-carbonyl compounds in 32–94% yield in a single laboratory operation. The intermediacy of o-QMs is supported by control experiments utilizing enolate precursors and conventional alkyl halides as competitive alkylating agents and the isolation of 1,5-dicarbonyl products resulting from conjugate additions that do not restore the aromatic system. PMID:25906358

  11. Michael Additions of Highly Basic Enolates to ortho-Quinone Methides.

    PubMed

    Lewis, Robert S; Garza, Christopher J; Dang, Ann T; Pedro, Te Kie A; Chain, William J

    2015-05-01

    A protocol by which ketone or ester enolates and ortho-quinone methides (o-QMs) are generated in situ in a single reaction flask from silylated precursors under the action of anhydrous fluoride is reported. The reaction partners are joined to give a variety of β-(2-hydroxyphenyl)-carbonyl compounds in 32-94% yield in a single laboratory operation. The intermediacy of o-QMs is supported by control experiments utilizing enolate precursors and conventional alkyl halides as competitive alkylating agents and the isolation of 1,5-dicarbonyl products resulting from conjugate additions that do not restore the aromatic system.

  12. High frequency transcutaneous electrical nerve stimulation with diphenidol administration results in an additive antiallodynic effect in rats following chronic constriction injury.

    PubMed

    Lin, Heng-Teng; Chiu, Chong-Chi; Wang, Jhi-Joung; Hung, Ching-Hsia; Chen, Yu-Wen

    2015-03-01

    The impact of coadministration of transcutaneous electrical nerve stimulation (TENS) and diphenidol is not well established. Here we estimated the effects of diphenidol in combination with TENS on mechanical allodynia and tumor necrosis factor-α (TNF-α) expression. Using an animal chronic constriction injury (CCI) model, the rat was estimated for evidence of mechanical sensitivity via von Frey hair stimulation and TNF-α expression in the sciatic nerve using the ELISA assay. High frequency (100Hz) TENS or intraperitoneal injection of diphenidol (2.0μmol/kg) was applied daily, starting on postoperative day 1 (POD1) and lasting for the next 13 days. We demonstrated that both high frequency TENS and diphenidol groups had an increase in mechanical withdrawal thresholds of 60%. Coadministration of high frequency TENS and diphenidol gives better results of paw withdrawal thresholds in comparison with high frequency TENS alone or diphenidol alone. Both diphenidol and coadministration of high frequency TENS with diphenidol groups showed a significant reduction of the TNF-α level compared with the CCI or HFS group (P<0.05) in the sciatic nerve on POD7, whereas the CCI or high frequency TENS group exhibited a higher TNF-α level than the sham group (P<0.05). Our resulting data revealed that diphenidol alone, high frequency TENS alone, and the combination produced a reduction of neuropathic allodynia. Both diphenidol and the combination of diphenidol with high frequency TENS inhibited TNF-α expression. A moderately effective dose of diphenidol appeared to have an additive effect with high frequency TENS. Therefore, multidisciplinary treatments could be considered for this kind of mechanical allodynia. PMID:25596445

  13. Identification of a sugar beet BvM14-MADS box gene through differential gene expression analysis of monosomic addition line M14.

    PubMed

    Ma, Chunquan; Wang, Yuguang; Wang, Yuting; Wang, Lifa; Chen, Sixue; Li, Haiying

    2011-11-01

    Monosomic addition line M14 carrying an additional chromosome 9 from Beta corolliflora Zosimovic ex Buttler was obtained through hybridization between the wild species B. corolliflora and a cultivated species Beta vulgaris L. var Saccharifera Alef. The M14 line showed diplosporic reproduction and stress tolerance. To identify differentially expressed genes in M14, a subtractive cDNA library was prepared by suppression subtractive hybridization (SSH) between M14 (2n=18+1) and B. vulgaris (2n=18). A total of 190 unique sequences were identified in the library and their putative functions were analyzed using Gene Ontology (GO). One of the genes, designated as BvM14-MADS box, encodes a MADS box transcription factor. It was cloned from M14 and over-expressed in transgenic tobacco plants. Interestingly, this gene was located on chromosome 2 of B. vulgaris, not on the additional chromosome 9. Overexpression of BvM14-MADS box led to significant phenotypic changes in tobacco. The differential expression of BvM14-MADS box gene in M14 may be caused by the interaction between the additional chromosome 9 from B. corolliflora and the B. vulgaris chromosomes in M14.

  14. Identification of a sugar beet BvM14-MADS box gene through differential gene expression analysis of monosomic addition line M14.

    PubMed

    Ma, Chunquan; Wang, Yuguang; Wang, Yuting; Wang, Lifa; Chen, Sixue; Li, Haiying

    2011-11-01

    Monosomic addition line M14 carrying an additional chromosome 9 from Beta corolliflora Zosimovic ex Buttler was obtained through hybridization between the wild species B. corolliflora and a cultivated species Beta vulgaris L. var Saccharifera Alef. The M14 line showed diplosporic reproduction and stress tolerance. To identify differentially expressed genes in M14, a subtractive cDNA library was prepared by suppression subtractive hybridization (SSH) between M14 (2n=18+1) and B. vulgaris (2n=18). A total of 190 unique sequences were identified in the library and their putative functions were analyzed using Gene Ontology (GO). One of the genes, designated as BvM14-MADS box, encodes a MADS box transcription factor. It was cloned from M14 and over-expressed in transgenic tobacco plants. Interestingly, this gene was located on chromosome 2 of B. vulgaris, not on the additional chromosome 9. Overexpression of BvM14-MADS box led to significant phenotypic changes in tobacco. The differential expression of BvM14-MADS box gene in M14 may be caused by the interaction between the additional chromosome 9 from B. corolliflora and the B. vulgaris chromosomes in M14. PMID:21807438

  15. Extremely Accessible Potassium Nitrate (KNO3) as the Highly Efficient Electrolyte Additive in Lithium Battery.

    PubMed

    Jia, Weishang; Fan, Cong; Wang, Liping; Wang, Qingji; Zhao, Mingjuan; Zhou, Aijun; Li, Jingze

    2016-06-22

    The systematic investigation of RNO3 salts (R = Li, Na, K, and Cs) as electrolyte additives was carried out for lithium-battery systems. For the first time, the abundant and extremely available KNO3 was proved to be an excellent alternative of LiNO3 for suppression of the lithium dendrites. The reason was ascribed to the possible synergetic effect of K(+) and NO3(-) ions: The positively charged K(+) ion could surround the lithium dendrites by electrostatic attraction and then delay their further growth, while simultaneously the oxidative NO3(-) ion could be reduced and subsequently profitable to the reinforcement of the solid-electrolyte interphase (SEI). By adding KNO3 into the practical Li-S battery, the discharging capacity was enhanced to average 687 mAh g(-1) from the case without KNO3 (528 mAh g(-1)) during 100 cycles, which was comparable to the one with the well-known LiNO3 additive (637 mAh g(-1)) under the same conditions. PMID:27237827

  16. High temperature properties of equiatomic FeAl with ternary additions

    NASA Technical Reports Server (NTRS)

    Titran, R. H.; Vedula, K. M.; Anderson, G. G.

    1984-01-01

    The aluminide intermetallic compounds are considered potential structural materials for aerospace applications. The B2 binary aluminide FeAl has a melting point in excess of 1500 K, is of simple cubic structure, exits over a wide range of composition with solubility for third elements and is potentially self-protecting in extreme environments. The B2 FeAl compound has been alloyed with 1 to 5 at % ternary additions of Si, Ti, Zr, Hf, Cr, Ni, Co, Nb, Ta, Mo, W, and Re. The alloys were prepared by blending a third elemental powder with prealloyed binary FeAl powder. Consolidation was by hot extrusion at 1250 K. Annealing studies on the extruded rods showed that the third element addition can be classified into three categories based upon the amount of homogenization and the extent of solid solutioning. Constant strain rate compression tests were performed to determine the flow stress as a function of temperature and composition. The mechanical strength behavior was dependent upon the third element homogenization classification.

  17. High temperature hydrogen sulfide adsorption on activated carbon - I. Effects of gas composition and metal addition

    USGS Publications Warehouse

    Cal, M.P.; Strickler, B.W.; Lizzio, A.A.

    2000-01-01

    Various types of activated carbon sorbents were evaluated for their ability to remove H2S from a simulated coal gas stream at a temperature of 550 ??C. The ability of activated carbon to remove H2S at elevated temperature was examined as a function of carbon surface chemistry (oxidation, thermal desorption, and metal addition), and gas composition. A sorbent prepared by steam activation, HNO3 oxidation and impregnated with Zn, and tested in a gas stream containing 0.5% H2S, 50% CO2 and 49.5% N2, had the greatest H2S adsorption capacity. Addition of H2, CO, and H2O to the inlet gas stream reduced H2S breakthrough time and H2S adsorption capacity. A Zn impregnated activated carbon, when tested using a simulated coal gas containing 0.5% H2S, 49.5% N2, 13% H2, 8.5% H2O, 21% CO, and 7.5% CO2, had a breakthrough time of 75 min, which was less than 25 percent of the length of breakthrough for screening experiments performed with a simplified gas mixture of 0.5% H2S, 50% CO2, and 49.5% N2.

  18. Viscoelastic properties of addition-cured polyimides used in high temperature polymer matrix composites

    NASA Technical Reports Server (NTRS)

    Roberts, Gary D; Malarik, Diane C.; Robaidek, Jerrold O.

    1991-01-01

    Viscoelastic properties of the addition cured polyimide, PMR-15, were studied using dynamic mechanical and stress relaxation tests. For temperatures below the glass transition temperature, T sub g, the dynamic mechanical properties measured using a temperature scan rate of 10 C/min were strongly affected by the presence of absorbed moisture in the resin. Dynamic mechanical properties measured as a function of time during an isothermal hold provided an indication of chemical changes occurring in the resin. For temperatures above (T sub g + 20 C), the storage modulus increased continuously as a function of time indicating that additional crosslinking is occurring in the resin. Because of these changes in chemical structures, the stress relaxation modulus could not be measured over any useful time interval for temperatures above T sub g. For temperatures below T sub g, dynamic mechanical properties appeared to be unaffected by chemical changes for times exceeding 1 hr. Since the duration of the stress relaxation tests was less than 1 hr, the stress relaxation modulus could be measured. As long as the moisture content of the resin was less than 2 pct, stress relaxation curves measured at different temperatures could be superimposed using horizontal shifts along the log(time) axis with only small shifts along the vertical axis.

  19. Extremely Accessible Potassium Nitrate (KNO3) as the Highly Efficient Electrolyte Additive in Lithium Battery.

    PubMed

    Jia, Weishang; Fan, Cong; Wang, Liping; Wang, Qingji; Zhao, Mingjuan; Zhou, Aijun; Li, Jingze

    2016-06-22

    The systematic investigation of RNO3 salts (R = Li, Na, K, and Cs) as electrolyte additives was carried out for lithium-battery systems. For the first time, the abundant and extremely available KNO3 was proved to be an excellent alternative of LiNO3 for suppression of the lithium dendrites. The reason was ascribed to the possible synergetic effect of K(+) and NO3(-) ions: The positively charged K(+) ion could surround the lithium dendrites by electrostatic attraction and then delay their further growth, while simultaneously the oxidative NO3(-) ion could be reduced and subsequently profitable to the reinforcement of the solid-electrolyte interphase (SEI). By adding KNO3 into the practical Li-S battery, the discharging capacity was enhanced to average 687 mAh g(-1) from the case without KNO3 (528 mAh g(-1)) during 100 cycles, which was comparable to the one with the well-known LiNO3 additive (637 mAh g(-1)) under the same conditions.

  20. Highly efficient "on water" catalyst-free nucleophilic addition reactions using difluoroenoxysilanes: dramatic fluorine effects.

    PubMed

    Yu, Jin-Sheng; Liu, Yun-Lin; Tang, Jing; Wang, Xin; Zhou, Jian

    2014-09-01

    A remarkable fluorine effect on "on water" reactions is reported. The CF⋅⋅⋅HO interactions between suitably fluorinated nucleophiles and the hydrogen-bond network at the phase boundary of oil droplets enable the formation of a unique microstructure to facilitate on water catalyst-free reactions, which are difficult to realize using nonfluorinated substrates. Accordingly, a highly efficient on water, catalyst-free reaction of difluoroenoxysilanes with aldehydes, activated ketones, and isatylidene malononitriles was developed, thus leading to the highly efficient synthesis of a variety of α,α-difluoro-β-hydroxy ketones and quaternary oxindoles.

  1. EVALUATION OF FGD DRY INJECTION SORBENTS AND ADDITIVES - VOLUME 1 - DEVELOPMENT OF HIGH REACTIVITY SORBENTS

    EPA Science Inventory

    The report discusses recent work addressing lime enhancement by slurrying with siliceous materials and testing in a laboratory packed-bed reactor, as part of EPA's efforts to develop low cost, retrofit flue gas cleaning technology, including the development of highly reactive sor...

  2. High-resolution and Monte Carlo additions to the SASKTRAN radiative transfer model

    NASA Astrophysics Data System (ADS)

    Zawada, D. J.; Dueck, S. R.; Rieger, L. A.; Bourassa, A. E.; Lloyd, N. D.; Degenstein, D. A.

    2015-06-01

    The Optical Spectrograph and InfraRed Imaging System (OSIRIS) instrument on board the Odin spacecraft has been measuring limb-scattered radiance since 2001. The vertical radiance profiles measured as the instrument nods are inverted, with the aid of the SASKTRAN radiative transfer model, to obtain vertical profiles of trace atmospheric constituents. Here we describe two newly developed modes of the SASKTRAN radiative transfer model: a high-spatial-resolution mode and a Monte Carlo mode. The high-spatial-resolution mode is a successive-orders model capable of modelling the multiply scattered radiance when the atmosphere is not spherically symmetric; the Monte Carlo mode is intended for use as a highly accurate reference model. It is shown that the two models agree in a wide variety of solar conditions to within 0.2 %. As an example case for both models, Odin-OSIRIS scans were simulated with the Monte Carlo model and retrieved using the high-resolution model. A systematic bias of up to 4 % in retrieved ozone number density between scans where the instrument is scanning up or scanning down was identified. The bias is largest when the sun is near the horizon and the solar scattering angle is far from 90°. It was found that calculating the multiply scattered diffuse field at five discrete solar zenith angles is sufficient to eliminate the bias for typical Odin-OSIRIS geometries.

  3. High resolution and Monte Carlo additions to the SASKTRAN radiative transfer model

    NASA Astrophysics Data System (ADS)

    Zawada, D. J.; Dueck, S. R.; Rieger, L. A.; Bourassa, A. E.; Lloyd, N. D.; Degenstein, D. A.

    2015-03-01

    The OSIRIS instrument on board the Odin spacecraft has been measuring limb scattered radiance since 2001. The vertical radiance profiles measured as the instrument nods are inverted, with the aid of the SASKTRAN radiative transfer model, to obtain vertical profiles of trace atmospheric constituents. Here we describe two newly developed modes of the SASKTRAN radiative transfer model: a high spatial resolution mode, and a Monte Carlo mode. The high spatial resolution mode is a successive orders model capable of modelling the multiply scattered radiance when the atmosphere is not spherically symmetric; the Monte Carlo mode is intended for use as a highly accurate reference model. It is shown that the two models agree in a wide variety of solar conditions to within 0.2%. As an example case for both models, Odin-OSIRIS scans were simulated with the Monte Carlo model and retrieved using the high resolution model. A systematic bias of up to 4% in retrieved ozone number density between scans where the instrument is scanning up or scanning down was identified. It was found that calculating the multiply scattered diffuse field at five discrete solar zenith angles is sufficient to eliminate the bias for typical Odin-OSIRIS geometries.

  4. Evaluation of alternative chemical additives for high-level waste vitrification feed preparation processing

    SciTech Connect

    Seymour, R.G.

    1995-06-07

    During the development of the feed processing flowsheet for the Defense Waste Processing Facility (DWPF) at the Savannah River Site (SRS), research had shown that use of formic acid (HCOOH) could accomplish several processing objectives with one chemical addition. These objectives included the decomposition of tetraphenylborate, chemical reduction of mercury, production of acceptable rheological properties in the feed slurry, and controlling the oxidation state of the glass melt pool. However, the DEPF research had not shown that some vitrification slurry feeds had a tendency to evolve hydrogen (H{sub 2}) and ammonia (NH{sub 3}) as the result of catalytic decomposition of CHOOH with noble metals (rhodium, ruthenium, palladium) in the feed. Testing conducted at Pacific Northwest Laboratory and later at the Savannah River Technical Center showed that the H{sub 2} and NH{sub 3} could evolve at appreciable rates and quantities. The explosive nature of H{sub 2} and NH{sub 3} (as ammonium nitrate) warranted significant mitigation control and redesign of both facilities. At the time the explosive gas evolution was discovered, the DWPF was already under construction and an immediate hardware fix in tandem with flowsheet changes was necessary. However, the Hanford Waste Vitrification Plant (HWVP) was in the design phase and could afford to take time to investigate flowsheet manipulations that could solve the problem, rather than a hardware fix. Thus, the HWVP began to investigate alternatives to using HCOOH in the vitrification process. This document describes the selection, evaluation criteria, and strategy used to evaluate the performance of the alternative chemical additives to CHOOH. The status of the evaluation is also discussed.

  5. Expression of Dominant-Negative Thyroid Hormone Receptor Alpha1 in Leydig and Sertoli Cells Demonstrates No Additional Defect Compared with Expression in Sertoli Cells Only

    PubMed Central

    Fumel, Betty; Froment, Pascal; Holzenberger, Martin; Livera, Gabriel; Monget, Philippe; Fouchécourt, Sophie

    2015-01-01

    Background In the testis, thyroid hormone (T3) regulates the number of gametes produced through its action on Sertoli cell proliferation. However, the role of T3 in the regulation of steroidogenesis is still controversial. Methods The TRαAMI knock-in allele allows the generation of transgenic mice expressing a dominant-negative TRα1 (thyroid receptor α1) isoform restricted to specific target cells after Cre-loxP recombination. Here, we introduced this mutant allele in both Sertoli and Leydig cells using a novel aromatase-iCre (ARO-iCre) line that expresses Cre recombinase under control of the human Cyp19(IIa)/aromatase promoter. Findings We showed that loxP recombination induced by this ARO-iCre is restricted to male and female gonads, and is effective in Sertoli and Leydig cells, but not in germ cells. We compared this model with the previous introduction of TRαAMI specifically in Sertoli cells in order to investigate T3 regulation of steroidogenesis. We demonstrated that TRαAMI-ARO males exhibited increased testis weight, increased sperm reserve in adulthood correlated to an increased proliferative index at P3 in vivo, and a loss of T3-response in vitro. Nevertheless, TRαAMI-ARO males showed normal fertility. This phenotype is similar to TRαAMI-SC males. Importantly, plasma testosterone and luteinizing hormone levels, as well as mRNA levels of steroidogenesis enzymes StAR, Cyp11a1 and Cyp17a1 were not affected in TRαAMI-ARO. Conclusions/Significance We concluded that the presence of a mutant TRαAMI allele in both Leydig and Sertoli cells does not accentuate the phenotype in comparison with its presence in Sertoli cells only. This suggests that direct T3 regulation of steroidogenesis through TRα1 is moderate in Leydig cells, and that Sertoli cells are the main target of T3 action in the testis. PMID:25793522

  6. Additional Development and Systems Analyses of Pneumatic Technology for High Speed Civil Transport Aircraft

    NASA Technical Reports Server (NTRS)

    Englar, Robert J.; Willie, F. Scott; Lee, Warren J.

    1999-01-01

    In the Task I portion of this NASA research grant, configuration development and experimental investigations have been conducted on a series of pneumatic high-lift and control surface devices applied to a generic High Speed Civil Transport (HSCT) model configuration to determine their potential for improved aerodynamic performance, plus stability and control of higher performance aircraft. These investigations were intended to optimize pneumatic lift and drag performance; provide adequate control and longitudinal stability; reduce separation flowfields at high angle of attack; increase takeoff/climbout lift-to-drag ratios; and reduce system complexity and weight. Experimental aerodynamic evaluations were performed on a semi-span HSCT generic model with improved fuselage fineness ratio and with interchangeable plain flaps, blown flaps, pneumatic Circulation Control Wing (CCW) high-lift configurations, plain and blown canards, a novel Circulation Control (CC) cylinder blown canard, and a clean cruise wing for reference. Conventional tail power was also investigated for longitudinal trim capability. Also evaluated was unsteady pulsed blowing of the wing high-lift system to determine if reduced pulsed mass flow rates and blowing requirements could be made to yield the same lift as that resulting from steady-state blowing. Depending on the pulsing frequency applied, reduced mass flow rates were indeed found able to provide lift augmentation at lesser blowing values than for the steady conditions. Significant improvements in the aerodynamic characteristics leading to improved performance and stability/control were identified, and the various components were compared to evaluate the pneumatic potential of each. Aerodynamic results were provided to the Georgia Tech Aerospace System Design Lab. to conduct the companion system analyses and feasibility study (Task 2) of theses concepts applied to an operational advanced HSCT aircraft. Results and conclusions from these

  7. Aberrant over-expression of COX-1 intersects multiple pro-tumorigenic pathways in high-grade serous ovarian cancer

    PubMed Central

    Wilson, Andrew J.; Fadare, Oluwole; Beeghly-Fadiel, Alicia; Son, Deok-Soo; Liu, Qi; Zhao, Shilin; Saskowski, Jeanette; Uddin, Md. Jashim; Daniel, Cristina; Crews, Brenda; Lehmann, Brian D.; Pietenpol, Jennifer A.; Crispens, Marta A.; Marnett, Lawrence J.; Khabele, Dineo

    2015-01-01

    Cyclooxygenase-1 (COX-1) is implicated in ovarian cancer. However, patterns of COX expression and function have been unclear and controversial. In this report, patterns of COX-1 and COX-2 gene expression were obtained from RNA-seq data through The Cancer Genome Atlas. Our analysis revealed markedly higher COX-1 mRNA expression than COX-2 in high-grade serous ovarian cancers (HGSOC) and higher COX-1 expression in HGSOC tumors than 10 other tumor types. High expression of COX-1 in HGSOC tumors was confirmed in an independent tissue microarray. In contrast, lower or similar expression of COX-1 compared to COX-2 was observed in endometrioid, mucinous and clear cell tumors. Stable COX-1 knockdown in HGSOC-representative OVCAR-3 ovarian cancer cells reduced gene expression in multiple pro-tumorigenic pathways. Functional cell viability, clonogenicity, and migration/invasion assays were consistent with transcriptomic changes. These effects were reversed by stable over-expression of COX-1 in SKOV-3 cells. Our results demonstrate a distinct pattern of COX-1 over-expression in HGSOC tumors and strong association of COX-1 with multiple pro-tumorigenic pathways in ovarian cancer cells. These findings provide additional insight into the role of COX-1 in human ovarian cancer and support further development of methods to selectively target COX-1 in the management of HGSOC tumors. PMID:25972361

  8. Enhanced critical currents in (Gd,Y)Ba2Cu3Ox superconducting tapes with high levels of Zr addition

    SciTech Connect

    Selvamanickam, V; Chen, Y; Shi, T; Liu, Y; Khatri, ND; Liu, J; Yao, Y; Xiong, X; Lei, C; Soloveichik, S; Galstyan, E; Majkic, G

    2013-01-21

    The critical current and structural properties of (Gd,Y)BaCuO tapes made by metal organic chemical vapor deposition (MOCVD) with Zr addition levels up to 30 at.% have been investigated. The reduction in critical current beyond the previously optimized Zr addition level of 7.5 at.% was found to be due to structural deterioration of the (Gd,Y)Ba2Cu3Ox film. By a modified MOCVD process,enhanced critical current densities have been achieved with high levels of Zr addition,including 3.83 MA cm(-2) in 15 at.% Zr- added 1.1 mu m thick film at 77 K in zero magnetic field. Critical currents as high as 1072 A/ 12 mm have been reached in (Gd,Y) BaCuO tapes with 15 at.% Zr addition at 30 K in a field of 3 T applied perpendicular to the tape,corresponding to a pinning force value of 268 GN m(-3). The enhanced critical currents achievable with a high density of nanoscale defects by employing high levels of second- phase additions enable the performance targets needed for the use of HTS tapes in coil applications involving high magnetic fields at temperatures below 50 K to be met.

  9. Enhanced critical currents in (Gd,Y)Ba2Cu3Ox superconducting tapes with high levels of Zr addition

    NASA Astrophysics Data System (ADS)

    Selvamanickam, V.; Chen, Y.; Shi, T.; Liu, Y.; Khatri, N. D.; Liu, J.; Yao, Y.; Xiong, X.; Lei, C.; Soloveichik, S.; Galstyan, E.; Majkic, G.

    2013-03-01

    The critical current and structural properties of (Gd,Y)BaCuO tapes made by metal organic chemical vapor deposition (MOCVD) with Zr addition levels up to 30 at.% have been investigated. The reduction in critical current beyond the previously optimized Zr addition level of 7.5 at.% was found to be due to structural deterioration of the (Gd,Y)Ba2Cu3Ox film. By a modified MOCVD process, enhanced critical current densities have been achieved with high levels of Zr addition, including 3.83 MA cm-2 in 15 at.% Zr-added 1.1 μm thick film at 77 K in zero magnetic field. Critical currents as high as 1072 A/12 mm have been reached in (Gd,Y)BaCuO tapes with 15 at.% Zr addition at 30 K in a field of 3 T applied perpendicular to the tape, corresponding to a pinning force value of 268 GN m-3. The enhanced critical currents achievable with a high density of nanoscale defects by employing high levels of second-phase additions enable the performance targets needed for the use of HTS tapes in coil applications involving high magnetic fields at temperatures below 50 K to be met.

  10. Snow survey and vegetation growth in high mountains (Swiss Alps) and additional ERTS investigations in Switzerland

    NASA Technical Reports Server (NTRS)

    Haefner, H. (Principal Investigator)

    1975-01-01

    The author has identified the following significant results. Two different methods, an analog and a digital one, have been developed for rapid and accurate mapping of the areal extent and changes in snow cover in high mountains. The quick-look method is based on individual visual control of each image using a photo quantizer which provides exact references for density slicing with high resolution lith-film. The digital snow classification system is based on discriminant analysis with the data of the four multispectral bands as variables and contains all preprocessing, feature extraction, and mapping steps for an operational application. Two different sets of sampling groups were established which apply to different conditions of snow cover. The first one serves for the normal situation with a uniform dry and new cover. The second one serves for situations with partly thawing and/or frozen snow.

  11. Clinoenstatite in alpe arami peridotite: additional evidence of very high pressure

    PubMed

    Bozhilov; Green; Dobrzhinetskaya

    1999-04-01

    Observations by transmission electron microscopy show that lamellae of clinoenstatite are present in diopside grains of the Alpe Arami garnet lherzolite of the Swiss Alps. The simplest interpretation of the orientation, crystallography, and microstructures of the lamellae and the phase relationships in this system is that the lamellae originally exsolved as the high-pressure C-centered form of clinoenstatite. These results imply that the rocks were exhumed from a minimum depth of 250 kilometers before or during continental collision. PMID:10102809

  12. Clinoenstatite in alpe arami peridotite: additional evidence of very high pressure

    PubMed

    Bozhilov; Green; Dobrzhinetskaya

    1999-04-01

    Observations by transmission electron microscopy show that lamellae of clinoenstatite are present in diopside grains of the Alpe Arami garnet lherzolite of the Swiss Alps. The simplest interpretation of the orientation, crystallography, and microstructures of the lamellae and the phase relationships in this system is that the lamellae originally exsolved as the high-pressure C-centered form of clinoenstatite. These results imply that the rocks were exhumed from a minimum depth of 250 kilometers before or during continental collision.

  13. A highly expressed miR-101 isomiR is a functional silencing small RNA

    PubMed Central

    2013-01-01

    Background MicroRNAs (miRNAs) are short non-coding regulatory RNAs that control gene expression usually producing translational repression and gene silencing. High-throughput sequencing technologies have revealed heterogeneity at length and sequence level for the majority of mature miRNAs (IsomiRs). Most isomiRs can be explained by variability in either Dicer1 or Drosha cleavage during miRNA biogenesis at 5’ or 3’ of the miRNA (trimming variants). Although isomiRs have been described in different tissues and organisms, their functional validation as modulators of gene expression remains elusive. Here we have characterized the expression and function of a highly abundant miR-101 5’-trimming variant (5’-isomiR-101). Results The analysis of small RNA sequencing data in several human tissues and cell lines indicates that 5’-isomiR-101 is ubiquitously detected and a highly abundant, especially in the brain. 5’-isomiR-101 was found in Ago-2 immunocomplexes and complementary approaches showed that 5’-isomiR-101 interacted with different members of the silencing (RISC) complex. In addition, 5’-isomiR-101 decreased the expression of five validated miR-101 targets, suggesting that it is a functional variant. Both the binding to RISC members and the degree of silencing were less efficient for 5’-isomiR-101 compared with miR-101. For some targets, both miR-101 and 5’-isomiR-101 significantly decreased protein expression with no changes in the respective mRNA levels. Although a high number of overlapping predicted targets suggest similar targeted biological pathways, a correlation analysis of the expression profiles of miR-101 variants and predicted mRNA targets in human brains at different ages, suggest specific functions for miR-101- and 5’-isomiR-101. Conclusions These results suggest that isomiRs are functional variants and further indicate that for a given miRNA, the different isomiRs may contribute to the overall effect as quantitative and

  14. Investigation of effect of particle size and rumen fluid addition on specific methane yields of high lignocellulose grass silage.

    PubMed

    Wall, D M; Straccialini, B; Allen, E; Nolan, P; Herrmann, C; O'Kiely, P; Murphy, J D

    2015-09-01

    This work examines the digestion of advanced growth stage grass silage. Two variables were investigated: particle size (greater than 3 cm and less than 1cm) and rumen fluid addition. Batch studies indicated particle size and rumen fluid addition had little effect on specific methane yields (SMYs). In continuous digestion of 3 cm silage the SMY was 342 and 343 L CH4 kg(-1)VS, respectively, with and without rumen fluid addition. However, digester operation was significantly affected through silage floating on the liquor surface and its entanglement in the mixing system. Digestion of 1cm silage with no rumen fluid addition struggled; volatile fatty acid concentrations rose and SMYs dropped. The best case was 1cm silage with rumen fluid addition, offering higher SMYs of 371 L CH4 kg(-1)VS and stable operation throughout. Thus, physical and biological treatments benefited continuous digestion of high fibre grass silage.

  15. Highly Functionalized Cyclopentane Derivatives by Tandem Michael Addition/Radical Cyclization/Oxygenation Reactions.

    PubMed

    Holan, Martin; Pohl, Radek; Císařová, Ivana; Klepetářová, Blanka; Jones, Peter G; Jahn, Ullrich

    2015-06-26

    Densely functionalized cyclopentane derivatives with up to four consecutive stereocenters are assembled by a tandem Michael addition/single-electron transfer oxidation/radical cyclization/oxygenation strategy mediated by ferrocenium hexafluorophosphate, a recyclable, less toxic single-electron transfer oxidant. Ester enolates were coupled with α-benzylidene and α-alkylidene β-dicarbonyl compounds with switchable diastereoselectivity. This pivotal steering element subsequently controls the diastereoselectivity of the radical cyclization step. The substitution pattern of the radical cyclization acceptor enables a switch of the cyclization mode from a 5-exo pattern for terminally substituted olefin units to a 6-endo mode for internally substituted acceptors. The oxidative anionic/radical strategy also allows efficient termination by oxygenation with the free radical 2,2,6,6-tetramethyl-1-piperidinoxyl, and two C-C bonds and one C-O bond are thus formed in the sequence. A stereochemical model is proposed that accounts for all of the experimental results and allows the prediction of the stereochemical outcome. Further transformations of the synthesized cyclopentanes are reported.

  16. High-dose nedocromil sodium as an addition to inhaled corticosteroids in the treatment of asthma.

    PubMed

    O'Hickey, S P; Rees, P J

    1994-08-01

    In a double-blind placebo-controlled trial nedocromil sodium in a dose of 8 mg four times daily or matching placebo was added to the treatment of 29 asthmatic patients. All patients were taking inhaled corticosteroids in a dose of up to 1000 micrograms daily. The trial agents were given for 6 weeks after a 2-week run-in period. Twenty-four patients completed the study, three withdrew because of adverse effects, two on placebo. Daytime asthma symptoms were significantly reduced on nedocromil compared to placebo (-0.46 vs. +0.09, P = 0.03). Night-time asthma and morning tightness were not changed significantly. Bronchodilator use in the night and day were lower on nedocromil but the differences were not significant. Morning peak flow rates were higher on nedocromil (+22.2 vs. +0.08, P = 0.06) and physicians opinions of overall effectiveness favoured nedocromil (U = 35.0, P = 0.04). These results confirm that nedocromil sodium may be a useful addition in asthma to low to medium doses of inhaled corticosteroids. The effects of 32 mg nedocromil daily were comparable to previous reports with lower doses.

  17. High-performance lubricant additives based on modified graphene oxide by ionic liquids.

    PubMed

    Fan, Xiaoqiang; Wang, Liping

    2015-08-15

    Graphene oxide (GO) is a layered material bearing a variety of oxygen-containing functional groups on its basal planes and edges, which allow it as a substrate to conduct a variety of chemical transformations. Here modified graphene oxide (MGO) was prepared using alkyl imidazolium ionic liquids (ILs) (1-butyl-3-methylimidazolium tetrafluoroborate (LB104), 1-butyl-3-methyl imidazolium hexafluorophosphate (LP104) and 1-hexyl-3-methylimidazolium bis(trifluoromethylsulfonyl) amide (LF106)) via epoxide ring-opening reaction, cation-π stacking or van der Waals interactions, with LB104 modified graphene (MG) exfoliated from graphite rod by a moderate electrochemical method as a comparison. The stability and tribological properties of MGO and MG as multialkylated cyclopentanes (MACs) additives were investigated in detail. The results show that GO is converted into graphene through the chemical modification using ILs, and MGO with good dispersion and stability in MACs significantly improves the tribological performance (friction and wear were reduced about 27% and 74% with pure MACs as a comparison, respectively). The excellent tribological properties are attributed to the formation of an ILs-containing graphene-rich tribofilm on the sliding surfaces, which as the third body can prevent the sliding surfaces from straight asperity contact and improve friction reducing and anti-wear behaviors.

  18. High-performance lubricant additives based on modified graphene oxide by ionic liquids.

    PubMed

    Fan, Xiaoqiang; Wang, Liping

    2015-08-15

    Graphene oxide (GO) is a layered material bearing a variety of oxygen-containing functional groups on its basal planes and edges, which allow it as a substrate to conduct a variety of chemical transformations. Here modified graphene oxide (MGO) was prepared using alkyl imidazolium ionic liquids (ILs) (1-butyl-3-methylimidazolium tetrafluoroborate (LB104), 1-butyl-3-methyl imidazolium hexafluorophosphate (LP104) and 1-hexyl-3-methylimidazolium bis(trifluoromethylsulfonyl) amide (LF106)) via epoxide ring-opening reaction, cation-π stacking or van der Waals interactions, with LB104 modified graphene (MG) exfoliated from graphite rod by a moderate electrochemical method as a comparison. The stability and tribological properties of MGO and MG as multialkylated cyclopentanes (MACs) additives were investigated in detail. The results show that GO is converted into graphene through the chemical modification using ILs, and MGO with good dispersion and stability in MACs significantly improves the tribological performance (friction and wear were reduced about 27% and 74% with pure MACs as a comparison, respectively). The excellent tribological properties are attributed to the formation of an ILs-containing graphene-rich tribofilm on the sliding surfaces, which as the third body can prevent the sliding surfaces from straight asperity contact and improve friction reducing and anti-wear behaviors. PMID:25935280

  19. Optimized Expression and Purification for High-Activity Preparations of Algal [FeFe]-Hydrogenase

    SciTech Connect

    Yacoby, I.; Tegler, L. T.; Pochekailov, S.; Zhang, S.; King, P. W.

    2012-04-01

    Recombinant expression and purification of metallo-enzymes, including hydrogenases, at high-yields is challenging due to complex, and enzyme specific, post-translational maturation processes. Low fidelities of maturation result in preparations containing a significant fraction of inactive, apo-protein that are not suitable for biophysical or crystallographic studies. We describe the construction, overexpression and high-yield purification of a fusion protein consisting of the algal [2Fe2S]-ferredoxin PetF (Fd) and [FeFe]-hydrogenase HydA1. The maturation of Fd-HydA1 was optimized through improvements in culture conditions and media components used for expression. We also demonstrated that fusion of Fd to the N-terminus of HydA1, in comparison to the C-terminus, led to increased expression levels that were 4-fold higher. Together, these improvements led to enhanced HydA1 activity and improved yield after purification. The strong binding-affinity of Fd for DEAE allowed for two-step purification by ion exchange and StrepTactin affinity chromatography. In addition, the incorporation of a TEV protease site in the Fd-HydA1 linker allowed for the proteolytic removal of Fd after DEAE step, and purification of HydA1 alone by StrepTactin. In combination, this process resulted in HydA1 purification yields of 5 mg L{sup -1} of culture from E. coli with specific activities of 1000 U (U = 1 {micro}mol hydrogen evolved mg{sup -1} min{sup -1}). The [FeFe]-hydrogenases are highly efficient enzymes and their catalytic sites provide model structures for synthetic efforts to develop robust hydrogen activation catalysts. In order to characterize their structure-function properties in greater detail, and to use hydrogenases for biotechnological applications, reliable methods for rapid, high-yield expression and purification are required.

  20. Adaptation of the pore diffusion model to describe multi-addition batch uptake high-throughput screening experiments.

    PubMed

    Traylor, Steven J; Xu, Xuankuo; Li, Yi; Jin, Mi; Li, Zheng Jian

    2014-11-14

    Equilibrium isotherm and kinetic mass transfer measurements are critical to mechanistic modeling of binding and elution behavior within a chromatographic column. However, traditional methods of measuring these parameters are impractically time- and labor-intensive. While advances in high-throughput robotic liquid handling systems have created time and labor-saving methods of performing kinetic and equilibrium measurements of proteins on chromatographic resins in a 96-well plate format, these techniques continue to be limited by physical constraints on protein addition, incubation and separation times; the available concentration of protein stocks and process pools; and practical constraints on resin and fluid volumes in the 96-well format. In this study, a novel technique for measuring protein uptake kinetics (multi-addition batch uptake) has been developed to address some of these limitations during high-throughput batch uptake kinetic measurements. This technique uses sequential additions of protein stock to chromatographic resin in a 96-well plate and the subsequent removal of each addition by centrifugation or vacuum separation. The pore diffusion model was adapted here to model multi-addition batch uptake and was tested and compared with traditional batch uptake measurements of uptake of an Fc-fusion protein on an anion exchange resin. Acceptable agreement between the two techniques is achieved for the two solution conditions investigated here. In addition, a sensitivity analysis of the model to the physical inputs is presented and the advantages and limitations of the multi-addition batch uptake technique are explored.

  1. Maternal High Fat Diet Affects Offspring’s Vitamin K-Dependent Proteins Expression Levels

    PubMed Central

    Lanham, Stuart; Cagampang, Felino R.; Oreffo, Richard O. C.

    2015-01-01

    Studies suggest bone growth & development and susceptibility to vascular disease in later life are influenced by maternal nutrition, during intrauterine and early postnatal life. There is evidence for a role of vitamin K-dependent proteins (VKDPs) including Osteocalcin, Matrix-gla protein, Periostin, and Gas6, in bone and vascular development. This study extends the analysis of VKDPs previously conducted in 6 week old offspring, into offspring of 30 weeks of age, to assess the longer term effects of a maternal and postnatal high fat (HF) diet on VKDP expression. Overall a HF maternal diet and offspring diet exacerbated the bone changes observed. Sex specific and tissue specific differences were observed in VKDP expression for both aorta and femoral tissues. In addition, significant correlations were observed between femoral OCN, Periostin Gas6, and Vkor expression levels and measures of femoral bone structure. Furthermore, MGP, OCN, Ggcx and Vkor expression levels correlated to mass and fat volume, in both sexes. In summary the current study has highlighted the importance of the long-term effects of maternal nutrition on offspring bone development and the correlation of VKDPs to bone structure. PMID:26381752

  2. Highly tissue specific expression of Sphinx supports its male courtship related role in Drosophila melanogaster.

    PubMed

    Chen, Ying; Dai, Hongzheng; Chen, Sidi; Zhang, Luoying; Long, Manyuan

    2011-01-01

    Sphinx is a lineage-specific non-coding RNA gene involved in regulating courtship behavior in Drosophila melanogaster. The 5' flanking region of the gene is conserved across Drosophila species, with the proximal 300 bp being conserved out to D. virilis and a further 600 bp region being conserved amongst the melanogaster subgroup (D. melanogaster, D. simulans, D. sechellia, D. yakuba, and D. erecta). Using a green fluorescence protein transformation system, we demonstrated that a 253 bp region of the highly conserved segment was sufficient to drive sphinx expression in male accessory gland. GFP signals were also observed in brain, wing hairs and leg bristles. An additional ∼800 bp upstream region was able to enhance expression specifically in proboscis, suggesting the existence of enhancer elements. Using anti-GFP staining, we identified putative sphinx expression signal in the brain antennal lobe and inner antennocerebral tract, suggesting that sphinx might be involved in olfactory neuron mediated regulation of male courtship behavior. Whole genome expression profiling of the sphinx knockout mutation identified significant up-regulated gene categories related to accessory gland protein function and odor perception, suggesting sphinx might be a negative regulator of its target genes.

  3. High expression of SGTA in esophageal squamous cell carcinoma correlates with proliferation and poor prognosis.

    PubMed

    Yang, Xiaojing; Cheng, Lei; Li, Mei; Shi, Hui; Ren, Hanru; Ding, Zongmei; Liu, Fang; Wang, Yuchan; Cheng, Chun

    2014-01-01

    Receptor tyrosine kinases (RTKs) expression and the growth factor such as platelet-derived growth factor (PDGF) and their receptors have been considered relevant in the process of angiogenesis and dissemination in esophageal squamous cell carcinoma (ESCC). Small glutamine-rich tetratricopeptide repeat-containing protein alpha (SGTA) downstream of RTK signaling was a critical regulator of PDGF receptors (PDGFR) stability. The aim of the present study was to examine the expression of SGTA and to elucidate its clinicopathologic significance in ESCC. Immunohistochemistry and western blot analysis were performed for SGTA in ESCC samples. SGTA was up-regulated in ESCC as compared with the adjacent normal tissue. High expression of SGTA was associated with tumor grade (P < 0.01), and SGTA was positively correlated with proliferation marker Ki-67 (P < 0.05). Univariate analysis showed that SGTA expression did has a remarkable prediction for poor prognosis (P = 0.016). Knockdown or overexpression of SGTA affected ESCC cells proliferation and cell cycle. Additionally, after ESCC cells silenced for SGTA were treated with cisplatin (an anti-ESCC agent), the cell growth was down-regulated. These findings suggested that SGTA was involved in the pathogenesis of ESCC and might indicate a poor prognosis for ESCC patients.

  4. High-Tg Thiol-Click Thermoset Networks via the Thiol-Maleimide Michael Addition.

    PubMed

    Parker, Shelbi; Reit, Radu; Abitz, Haley; Ellson, Gregory; Yang, Kejia; Lund, Benjamin; Voit, Walter E

    2016-07-01

    Thiol-click reactions lead to polymeric materials with a wide range of interesting mechanical, electrical, and optical properties. However, this reaction mechanism typically results in bulk materials with a low glass transition temperature (Tg ) due to rotational flexibility around the thioether linkages found in networks such as thiol-ene, thiol-epoxy, and thiol-acrylate systems. This report explores the thiol-maleimide reaction utilized for the first time as a solvent-free reaction system to synthesize high-Tg thermosetting networks. Through thermomechanical characterization via dynamic mechanical analysis, the homogeneity and Tg s of thiol-maleimide networks are compared to similarly structured thiol-ene and thiol-epoxy networks. While preliminary data show more heterogeneous networks for thiol-maleimide systems, bulk materials exhibit Tg s 80 °C higher than other thiol-click systems explored herein. Finally, hollow tubes are synthesized using each thiol-click reaction mechanism and employed in low- and high-temperature environments, demonstrating the ability to withstand a compressive radial 100 N deformation at 100 °C wherein other thiol-click systems fail mechanically. PMID:27168131

  5. Mobile hard substrata - An additional biodiversity source in a high latitude shallow subtidal system

    NASA Astrophysics Data System (ADS)

    Balazy, Piotr; Kuklinski, Piotr

    2013-03-01

    This study demonstrates the importance of a hard mobile substratum (hermit crab shells) for Arctic biodiversity. Based on previous observations from other geographic regions we hypothesized that this niche at high latitudes would support a higher biodiversity of epifauna than might be predicted from similar substrata. We test whether the hermit crab epifauna is specific to that substratum providing unique biodiversity components to the local community. From four study sites in Isfjorden (78°N), West Spitsbergen and two study sites in Northern Norway (69°N) we collected approximately 50 each of hermit crabs, gastropods and pebbles, of visually similar surface area using SCUBA diving. Hermit crab shells were colonized by a larger number of epifaunal species than either gastropods or pebbles, even when they were of a larger size. Among 87 taxa found on all the three substrata, 22 occurred only on hermit crab shells. Except for two study sites hermit crab shells also supported more individuals. This study shows that the contribution of shells carried by hermit crabs to high-latitude, shallow-subtidal diversity is higher than might be predicted by their surface area alone and that hermit crabs modify, maintain and create a unique habitat. This is the result of a number of factors interacting positively on the presence of epifauna including shell surface heterogeneity and the complex influence of the crab host.

  6. HIGH YIELD SOLUBLE BACTERIAL EXPRESSION AND STREAMLINED PURIFICATION OF RECOMBINANT HUMAN INTERFERON α-2A

    PubMed Central

    Bis, Regina L.; Stauffer, Tara M.; Singh, Surinder M.; Lavoie, Thomas B.; Mallela, Krishna M.G.

    2014-01-01

    Interferon α-2a (IFNA2) is a member of the Type I interferon cytokine family, known for its antiviral and anti-proliferative functions. The role of this family in the innate immune response makes it an attractive candidate for the treatment of many viral and chronic immune-compromised diseases. Recombinant IFNA2 is clinically used to modulate hairy cell leukemia as well as hepatitis c. Historically, IFNA2 has been purified from human leukocytes as well as bacterial expression systems. In most cases, bacterial expression of IFNA2 resulted in inclusion body formation, or required numerous purification steps that decreased the protein yield. Here, we describe an expression and purification scheme for IFNA2 using a pET-SUMO bacterial expression system and a single purification step. Using SUMO protein as the fusion tag achieved high soluble protein expression. The SUMO tag was cleaved with the Ulp1 protease leaving no additional amino acids on the fusion terminus following the cleavage. Mass spectrometry, circular dichroism, 2D heteronuclear NMR, and analytical ultracentrifugation confirmed the amino acid sequence identity, secondary and tertiary protein structures, and the solution behavior of the purified IFNA2. The purified protein also has antiviral and anti-proliferative activities comparable to the WHO International Standard, NIBSC 95/650, and the IFNA2 standard available from PBL Assay Science. Combining the expression and purification protocols developed here to produce IFNA2 on a laboratory scale with the commercial fermenter technology commonly used in pharmaceutical industry may further enhance IFNA2 yields, which will promote the development of interferon-based protein drugs to treat various disorders. PMID:24794500

  7. Impact of electrolyte solvent and additive choices on high voltage Li-ion pouch cells

    NASA Astrophysics Data System (ADS)

    Xia, Jian; Nelson, K. J.; Lu, Zhonghua; Dahn, J. R.

    2016-10-01

    The effects that various electrolyte solvents and electrolyte additives had on both LaPO4-coated LiNi0.4Mn0.4Co0.2O2 and uncoated LiNi0.4Mn0.4Co0.2O2/graphite pouch cells were studied using automated storage, electrochemical impedance spectroscopy, gas production and long-term cycling experiments. Storage experiments showed that the voltage drop during storage at 4.3 or 4.4 V for both coated and uncoated cells was very similar for the same electrolyte choice. At 4.5 V or above, the LaPO4-coated cells had a significantly smaller voltage drop than the uncoated cells except when fluorinated electrolytes were used. Automated charge discharge cycling/impedance spectroscopy testing of cells held at 4.5 V for 24 h every cycle showed that all cells containing ethylene carbonate:ethyl methyl carbonate electrolyte or sulfolane:ethyl methyl carbonate electrolyte exhibited severe capacity fade. By contrast, cells containing fluorinated electrolytes had the best capacity retention and smallest impedance growth during these aggressive cycling/hold tests. Long-term cycling experiments to 4.5 V confirmed that cells containing fluorinated electrolyte had the best cycling performance in the uncoated LiNi0.4Mn0.4Co0.2O2/graphite cells while cells containing sulfolane:ethyl methyl carbonate electrolyte had the best cycling performance in coated LiNi0.4Mn0.4Co0.2O2/graphite cells.

  8. Anaerobic digestion of high-strength cheese whey utilizing semicontinuous digesters and chemical flocculant addition

    SciTech Connect

    Barford, J.P.; Cail, R.G.; Callander, I.J.; Floyd, E.J.

    1986-11-01

    Semicontinuous digesters were used to anaerobically treat high-strength whey (70 kg/cubic m COD). A maximum loading of 16.1 kg COD/cubic m/day was obtained with soluble COD removal efficiencies greater than 99%. The use of a chemical flocculant resulted in an increased biomass concentration in the digester compared to a control, thus enabling correspondingly higher space loadings to be applied. With the onset of substantial levels of granulation of the biomass, flocculant dosage was able to be discontinued. This article discusses the performance of the digesters in detail and, briefly, the long-term operational difficulties experienced and the control strategies employed on such systems. 24 references.

  9. Composite materials based on high-modulus compounds for additive technology

    NASA Astrophysics Data System (ADS)

    Grigoriev, M.; Kotelnikov, N.; Buyakova, S.; Kulkov, S.

    2016-07-01

    The effect of adding nanocrystalline ZrO2 and submicron TiC to ultrafine Al2O3 on mechanical properties and the microstructure of the composites developed by hot pressing was investigated. It was shown that by means of hot pressing in argon atmosphere at the sintering temperature of 1500 °C one can obtain the composites of Al2O3-ZrO2-TiC with a fine structure and minimal porosity. It was shown that in the material a multi-scale hierarchical structure is formed, which possesses high physical and mechanical properties: the hardness and fracture toughness was 22 GPa and 5.2 MPa*m1/2, respectively. It has been shown that mechanical properties of the composite are better than those of commercial composites based on aluminum oxide (Al2O3, ZTA, Al2O3-TiC) and are comparable to those of silicon nitride.

  10. High-dose inhaled corticosteroids or addition of theophylline in patients with poorly controlled asthma?

    PubMed

    Celis, Pilar; Rada, Gabriel

    2015-08-19

    There are several management strategies for patients with poorly controlled asthma despite usual treatment. Increasing doses of inhaled corticosteroids or adding theophylline are among the therapeutic alternatives. However, the latter is associated with important adverse effects. Searching in Epistemonikos database, which is maintained by screening 30 databases, we identified only one systematic review including four pertinent randomized controlled trials. We combined the evidence using meta-analysis and generated a summary of findings following the GRADE approach. We concluded it is not clear whether theophylline or high-dose inhaled corticosteroids constitute a better alternative for symptomatic control or reduction in exacerbations in poorly controlled asthmatic patients because the certainty of the evidence is very low.

  11. Lateralization of High-Frequency Clicks Based on Interaural Time: Additivity of Information across Frequency

    NASA Astrophysics Data System (ADS)

    Wenzel, Elizabeth Marie

    Lateralization performance based on interaural differences of time (IDTs) was measured for trains of Gaussian clicks which varied in spectral content. In the first experiment, thresholds ((DELTA)IDTs) were measured as a function of the number of clicks in the train (n = 1 to 32), the interclick interval (ICI = 2.5 or 5 ms), and the spectral content (1 vs. 2 or 4 carriers). Subjects' performance was compared to perfect statistical summation which predicts slopes of -.50 when log-(DELTA)IDT vs. long -n is plotted. The results showed that increasing the spectral content of the clicks decreased the intercepts of the log -log functions (decreased thresholds) while having little effect on their slopes. Shortening the ICIs caused the slopes of the functions to decrease in absolute value. To estimate the bandwidth of frequency-interaction in lateralization, d's were measured for clicks with constant IDTs (n = 1) with a fixed carrier (FF = 4000, 5200, 6000 or 7200 Hz), both alone and combined with a second click whose carrier (F) varied from 3500 to 8500 Hz. Performance in combined conditions was compared to independent summation of the information carried by the two frequency-bands. Performance improved as the separation between F and FF increased until the level predicted by independence was reached. The final experiment investigated the interaction of frequency content with IDT. d's were measured as a function of the IDT in clicks with carriers of 5200, 6000 or 7200 Hz, both alone and combined with a 4000-Hz click with a fixed IDT. Performance in combined conditions was again compared to independent additivity. The improvement with frequency was explained by an increase in the number of samples of the IDT reaching the binaural centers due to spread of excitation along the basilar membrane. Less than independent summation was explained by correlation between overlapping bands which reduced the amount of information exciting independent channels. The data also suggest that

  12. A green chemistry approach to a more efficient asymmetric catalyst: solvent-free and highly concentrated alkyl additions to ketones.

    PubMed

    Jeon, Sang-Jin; Li, Hongmei; Walsh, Patrick J

    2005-11-30

    There is a great demand for development of catalyst systems that are not only efficient and highly enantioselective but are also environmentally benign. Herein we report investigations into the catalytic asymmetric addition of alkyl and functionalized alkyl groups to ketones under highly concentrated and solvent-free conditions. In comparison with standard reaction conditions employing toluene and hexanes, the solvent-free and highly concentrated conditions permit reduction in catalyst loading by a factor of 2- to 40-fold. These new conditions are general and applicable to a variety of ketones and dialkylzinc reagents to provide diverse tertiary alcohols with high enantioselectivities. Using cyclic conjugated enones, we have performed a tandem asymmetric addition/diastereoselective epoxidation using the solvent-free addition conditions followed by introduction of a 5.5 M decane solution of tert-butyl hydroperoxide (TBHP) to generate epoxy alcohols. This one-pot procedure allows access to syn epoxy alcohols with three contiguous stereocenters with excellent enantio- and diastereoselectivities and high yields. Both the solvent-free asymmetric additions and asymmetric addition/diastereoselective epoxidation reactions have been conducted on larger scale (5 g substrate) with 0.5 mol % catalyst loadings. In these procedures, enantioselectivities equal to or better than 92% were obtained with isolated yields of 90%. The solvent-free and highly concentrated conditions are a significant improvement over previous solvent-based protocols. Further, this chemistry represents a rare example of a catalytic asymmetric reaction that is highly enantioselective under more environmentally friendly solvent-free conditions. PMID:16305227

  13. Pregnant rat uterus expresses high levels of the type 3 iodothyronine deiodinase

    PubMed Central

    Galton, Valerie Anne; Martinez, Elena; Hernandez, Arturo; St. Germain, Emily A.; Bates, Joanne M.; St. Germain, Donald L.

    1999-01-01

    Although thyroid hormones are critically important for the coordination of morphogenic processes in the fetus and neonate, premature exposure of the embryo to levels of the hormones present in the adult is detrimental and can result in growth retardation, malformations, and even death. We report here that the pregnant rat uterus expresses extremely high levels of the type 3 iodothyronine deiodinase (D3), which inactivates thyroxine and 3,3′,5-triiodothyronine by 5-deiodination. Both D3 mRNA and activity were present at the implantation site as early as gestational day 9 (E9), when expression was localized using in situ hybridization to uterine mesometrial and antimesometrial decidual tissue. At later stages of gestation, uterine D3 activity remained very high, and the levels exceeded those observed in the placenta and in fetal tissues. After days E12 and E13, as decidual tissues regressed, D3 expression became localized to the epithelial cells lining the recanalized uterine lumen that surrounds the fetal cavity. These findings strongly suggest that the pregnant uterus, in addition to the placenta, plays a critical role in determining the level of exposure of the fetus to maternal thyroid hormones. J. Clin. Invest. 103:979–987 (1999). PMID:10194470

  14. Low expression of PKCα and high expression of KRAS predict poor prognosis in patients with colorectal cancer

    PubMed Central

    Chen, Suxian; Wang, Yadi; Zhang, Yun; Wan, Yizeng

    2016-01-01

    The current study aimed to determine the association between protein kinase Cα (PKCα) and Kirsten rat sarcoma viral oncogene homolog (KRAS) expression and the response to folinic acid, 5-fluorouracil and oxaliplatin (FOLFOX regimen) in patients with colorectal cancer (CRC). The protein levels of PKCα and KRAS were analyzed by immunohistochemistry in tissue samples from patients with CRC and in non-cancerous tissues, including 152 cases of colorectal adenocarcinoma, 30 cases of colorectal adenoma and 20 normal colonic mucosa samples. The association between PKCα and KRAS expression and clinicopathological features was analyzed. The rates of positive PKCα protein expression in patients with poorly, moderately and well-differentiated adenocarcinoma were 16.7% (6/36), 40.0% (24/60), and 57.1% (32/56), respectively (P<0.013). The rate of positive KRAS expression in CRC patients was significantly higher than in patients with colon adenoma and normal colon mucosa (P<0.001). Expression levels of KRAS were associated with the degree of differentiation of CRC (P<0.001). Expression of PKCα was negatively correlated with KRAS expression in CRC tissues. The mean progression-free survival (PFS) times in patients with high and low expression of PKCα were 43.9 and 38.8 months, respectively (P<0.001). The mean PFS times were 38.5 and 45.5 months in patients with high and low expression of KRAS, respectively (P=0.001). In conclusion, low PKCα and high KRAS expression predicted relatively poor prognosis in patients with CRC.

  15. Low expression of PKCα and high expression of KRAS predict poor prognosis in patients with colorectal cancer

    PubMed Central

    Chen, Suxian; Wang, Yadi; Zhang, Yun; Wan, Yizeng

    2016-01-01

    The current study aimed to determine the association between protein kinase Cα (PKCα) and Kirsten rat sarcoma viral oncogene homolog (KRAS) expression and the response to folinic acid, 5-fluorouracil and oxaliplatin (FOLFOX regimen) in patients with colorectal cancer (CRC). The protein levels of PKCα and KRAS were analyzed by immunohistochemistry in tissue samples from patients with CRC and in non-cancerous tissues, including 152 cases of colorectal adenocarcinoma, 30 cases of colorectal adenoma and 20 normal colonic mucosa samples. The association between PKCα and KRAS expression and clinicopathological features was analyzed. The rates of positive PKCα protein expression in patients with poorly, moderately and well-differentiated adenocarcinoma were 16.7% (6/36), 40.0% (24/60), and 57.1% (32/56), respectively (P<0.013). The rate of positive KRAS expression in CRC patients was significantly higher than in patients with colon adenoma and normal colon mucosa (P<0.001). Expression levels of KRAS were associated with the degree of differentiation of CRC (P<0.001). Expression of PKCα was negatively correlated with KRAS expression in CRC tissues. The mean progression-free survival (PFS) times in patients with high and low expression of PKCα were 43.9 and 38.8 months, respectively (P<0.001). The mean PFS times were 38.5 and 45.5 months in patients with high and low expression of KRAS, respectively (P=0.001). In conclusion, low PKCα and high KRAS expression predicted relatively poor prognosis in patients with CRC. PMID:27602102

  16. High expression of MAP7 predicts adverse prognosis in young patients with cytogenetically normal acute myeloid leukemia

    PubMed Central

    Fu, Lin; Fu, Huaping; Zhou, Lei; Xu, Keman; Pang, Yifan; Hu, Kai; Wang, Jing; Tian, Lei; Liu, Yuanyuan; Wang, Jijun; Jing, Hongmei; Huang, Wenrong; Ke, Xiaoyan; Shi, Jinlong

    2016-01-01

    Microtubule-associated protein 7 (MAP7) plays an important role in cancer cells. In this study, we identified the prognostic significance of MAP7 expression in cytogenetically normal acute myeloid leukemia (CN-AML) patients (aged <60 years) based on several microarray datasets. In the first group (n = 129), high MAP7 expression (MAP7high) was associated with adverse overall survival (OS; P = 0.0441) and event-free survival (EFS; P = 0.0114) compared with low MAP7 expression (MAP7low). In addition, the prognostic significance of MAP7 was confirmed by European Leukemia Net (ELN) intermediate-I genetic categories and multivariable analysis. In the second independent group of CN-AML patients (aged <60 years), MAP7high was also associated with adverse OS (n = 88, OS; P = 0.00811). To understand the inherent mechanisms of MAP7’s prognosis, we investigated genome-wide gene/microRNA expression signatures associated with MAP7 expression. Several known oncogenic genes/microRNAs and anti-oncogenic genes/microRNAs were disordered in MAP7high CN-AML patients. In conclusion, MAP7high is an adverse prognostic biomarker for CN-AML, which may be attributed to the distinctive genome-wide gene/microRNA expression and related cell signaling pathways. PMID:27686215

  17. Cell types differ in global coordination of splicing and proportion of highly expressed genes.

    PubMed

    Trakhtenberg, Ephraim F; Pho, Nam; Holton, Kristina M; Chittenden, Thomas W; Goldberg, Jeffrey L; Dong, Lingsheng

    2016-01-01

    Balance in the transcriptome is regulated by coordinated synthesis and degradation of RNA molecules. Here we investigated whether mammalian cell types intrinsically differ in global coordination of gene splicing and expression levels. We analyzed RNA-seq transcriptome profiles of 8 different purified mouse cell types. We found that different cell types vary in proportion of highly expressed genes and the number of alternatively spliced transcripts expressed per gene, and that the cell types that express more variants of alternatively spliced transcripts per gene are those that have higher proportion of highly expressed genes. Cell types segregated into two clusters based on high or low proportion of highly expressed genes. Biological functions involved in negative regulation of gene expression were enriched in the group of cell types with low proportion of highly expressed genes, and biological functions involved in regulation of transcription and RNA splicing were enriched in the group of cell types with high proportion of highly expressed genes. Our findings show that cell types differ in proportion of highly expressed genes and the number of alternatively spliced transcripts expressed per gene, which represent distinct properties of the transcriptome and may reflect intrinsic differences in global coordination of synthesis, splicing, and degradation of RNA molecules. PMID:27577089

  18. Cell types differ in global coordination of splicing and proportion of highly expressed genes

    PubMed Central

    Trakhtenberg, Ephraim F.; Pho, Nam; Holton, Kristina M.; Chittenden, Thomas W.; Goldberg, Jeffrey L.; Dong, Lingsheng

    2016-01-01

    Balance in the transcriptome is regulated by coordinated synthesis and degradation of RNA molecules. Here we investigated whether mammalian cell types intrinsically differ in global coordination of gene splicing and expression levels. We analyzed RNA-seq transcriptome profiles of 8 different purified mouse cell types. We found that different cell types vary in proportion of highly expressed genes and the number of alternatively spliced transcripts expressed per gene, and that the cell types that express more variants of alternatively spliced transcripts per gene are those that have higher proportion of highly expressed genes. Cell types segregated into two clusters based on high or low proportion of highly expressed genes. Biological functions involved in negative regulation of gene expression were enriched in the group of cell types with low proportion of highly expressed genes, and biological functions involved in regulation of transcription and RNA splicing were enriched in the group of cell types with high proportion of highly expressed genes. Our findings show that cell types differ in proportion of highly expressed genes and the number of alternatively spliced transcripts expressed per gene, which represent distinct properties of the transcriptome and may reflect intrinsic differences in global coordination of synthesis, splicing, and degradation of RNA molecules. PMID:27577089

  19. Characterization of Steel-Ta Dissimilar Metal Builds Made Using Very High Power Ultrasonic Additive Manufacturing (VHP-UAM)

    NASA Astrophysics Data System (ADS)

    Sridharan, Niyanth; Norfolk, Mark; Babu, Sudarsanam Suresh

    2016-05-01

    Ultrasonic additive manufacturing is a solid-state additive manufacturing technique that utilizes ultrasonic vibrations to bond metal tapes into near net-shaped components. The major advantage of this process is the ability to manufacture layered structures with dissimilar materials without any intermetallic formation. Majority of the published literature had focused only on the bond formation mechanism in Aluminum alloys. The current work pertains to explain the microstructure evolution during dissimilar joining of iron and tantalum using very high power ultrasonic additive manufacturing and characterization of the interfaces using electron back-scattered diffraction and Nano-indentation measurement. The results showed extensive grain refinement at the bonded interfaces of these metals. This phenomenon was attributed to continuous dynamic recrystallization process driven by the high strain rate plastic deformation and associated adiabatic heating that is well below 50 pct of melting point of both iron and Ta.

  20. Fatty Acid Profiles and Stearoyl-CoA Desaturase Gene Expression in Longissimus dorsi Muscle of Growing Lambs Influenced by Addition of Tea Saponins and Soybean Oil

    PubMed Central

    Mao, H. L.; Wang, J. K.; Lin, J.; Liu, J. X.

    2012-01-01

    This study was conducted to determine the effects of dietary addition of tea saponins (TS) and soybean oil (SO) on fatty acid profile and gene expression of stearoyl-CoA desaturase (SCD) in longissimus dorsi (LD) muscle of growing lambs. Thirty-two Huzhou lambs were assigned to four dietary treatments in a 2×2 factorial arrangement with main effects of TS (0 or 3 g/d) and SO (0 or 30 g/kg of diet DM). The diet without additives was considered as NTNS (no TS or SO). After a feeding trial for 60 d, four lambs of each treatment were slaughtered to collect the samples of LD muscle. Percentage of trans-11 vaccenic acid was enhanced (p<0.05) in muscle of lambs fed TS and SO. The proportion of total conjugated linoleic acid (CLA) was increased (p<0.05) by SO, but decreased (p<0.05) by TS in LD muscle. The percentage of total saturated fatty acids in muscle was decreased (p<0.05) by addition of TS and SO, while addition of SO increased (p<0.05) the percentage of total polyunsaturated fatty acids. The ratio of cis-9, trans-11 CLA to tran-11 vaccenic acid was decreased (p<0.05) by TS, but increased (p<0.05) by SO. The same effects were observed in SCD mRNA expression. From these results it is indicated that including TS and SO in the diet of growing lambs affect the fatty acid profiles of LD muscle and that the proportion of cis-9, trans-11 CLA in the muscle influenced by TS and SO may be related to the SCD gene expression. PMID:25049609

  1. Highly enantioselective Michael addition reactions of 2-substituted benzofuran-3(2H)-ones to nitroolefins.

    PubMed

    Zhang, Zhi-Pei; Dong, Nan; Li, Xin; Cheng, Jin-Pei

    2015-10-21

    A highly enantioselective Michael addition reaction of 2-substituted benzofuran-3(2H)-ones to nitroolefins was promoted by a bifunctional squaramide catalyst. As a result, a number of chiral 2,2'-substituted benzofuran-3-one derivatives, bearing adjacent quaternary-tertiary stereocenters, were efficiently synthesized with excellent enantioselectivities.

  2. Upward shift of the vortex solid phase in high-temperature-superconducting wires through high density nanoparticle addition.

    PubMed

    Miura, Masashi; Maiorov, Boris; Balakirev, Fedor F; Kato, Takeharu; Sato, Michio; Takagi, Yuji; Izumi, Teruo; Civale, Leonardo

    2016-02-08

    We show a simple and effective way to improve the vortex irreversibility line up to very high magnetic fields (60T) by increasing the density of second phase BaZrO3 nanoparticles. (Y0.77,Gd0.23)Ba2Cu3Oy films were grown on metal substrates with different concentration of BaZrO3 nanoparticles by the metal organic deposition method. We find that upon increase of the BaZrO3 concentration, the nanoparticle size remains constant but the twin-boundary density increases. Up to the highest nanoparticle concentration (n ~ 1.3 × 10(22)/m(3)), the irreversibility field (Hirr) continues to increase with no sign of saturation up to 60 T, although the vortices vastly outnumber pinning centers. We find extremely high Hirr, namely Hirr = 30 T (H||45°) and 24 T (H||c) at 65 K and 58 T (H||45°) and 45 T (H||c) at 50K. The difference in pinning landscape shifts the vortex solid-liquid transition upwards, increasing the vortex region useful for power applications, while keeping the upper critical field, critical temperature and electronic mass anisotropy unchanged.

  3. Upward shift of the vortex solid phase in high-temperature-superconducting wires through high density nanoparticle addition

    DOE PAGES

    Miura, Masashi; Maiorov, Boris; Balakirev, Fedor F.; Kato, Takeharu; Sato, Michio; Takagi, Yuji; Izumi, Teruo; Civale, Leonardo

    2016-02-08

    Here, we show a simple and effective way to improve the vortex irreversibility line up to very high magnetic fields (60T) by increasing the density of second phase BaZrO3 nanoparticles. (Y0.77,Gd0.23)Ba2Cu3Oy films were grown on metal substrates with different concentration of BaZrO3 nanoparticles by the metal organic deposition method. We find that upon increase of the BaZrO3 concentration, the nanoparticle size remains constant but the twin-boundary density increases. Up to the highest nanoparticle concentration (n ~ 1.3 × 1022/m3), the irreversibility field (Hirr) continues to increase with no sign of saturation up to 60 T, although the vortices vastly outnumbermore » pinning centers. We find extremely high Hirr, namely Hirr = 30 T (H||45°) and 24 T (H||c) at 65 K and 58 T (H||45°) and 45 T (H||c) at 50K. The difference in pinning landscape shifts the vortex solid-liquid transition upwards, increasing the vortex region useful for power applications, while keeping the upper critical field, critical temperature and electronic mass anisotropy unchanged.« less

  4. Upward shift of the vortex solid phase in high-temperature-superconducting wires through high density nanoparticle addition.

    PubMed

    Miura, Masashi; Maiorov, Boris; Balakirev, Fedor F; Kato, Takeharu; Sato, Michio; Takagi, Yuji; Izumi, Teruo; Civale, Leonardo

    2016-01-01

    We show a simple and effective way to improve the vortex irreversibility line up to very high magnetic fields (60T) by increasing the density of second phase BaZrO3 nanoparticles. (Y0.77,Gd0.23)Ba2Cu3Oy films were grown on metal substrates with different concentration of BaZrO3 nanoparticles by the metal organic deposition method. We find that upon increase of the BaZrO3 concentration, the nanoparticle size remains constant but the twin-boundary density increases. Up to the highest nanoparticle concentration (n ~ 1.3 × 10(22)/m(3)), the irreversibility field (Hirr) continues to increase with no sign of saturation up to 60 T, although the vortices vastly outnumber pinning centers. We find extremely high Hirr, namely Hirr = 30 T (H||45°) and 24 T (H||c) at 65 K and 58 T (H||45°) and 45 T (H||c) at 50K. The difference in pinning landscape shifts the vortex solid-liquid transition upwards, increasing the vortex region useful for power applications, while keeping the upper critical field, critical temperature and electronic mass anisotropy unchanged. PMID:26853703

  5. Upward shift of the vortex solid phase in high-temperature-superconducting wires through high density nanoparticle addition

    PubMed Central

    Miura, Masashi; Maiorov, Boris; Balakirev, Fedor F.; Kato, Takeharu; Sato, Michio; Takagi, Yuji; Izumi, Teruo; Civale, Leonardo

    2016-01-01

    We show a simple and effective way to improve the vortex irreversibility line up to very high magnetic fields (60T) by increasing the density of second phase BaZrO3 nanoparticles. (Y0.77,Gd0.23)Ba2Cu3Oy films were grown on metal substrates with different concentration of BaZrO3 nanoparticles by the metal organic deposition method. We find that upon increase of the BaZrO3 concentration, the nanoparticle size remains constant but the twin-boundary density increases. Up to the highest nanoparticle concentration (n ~ 1.3 × 1022/m3), the irreversibility field (Hirr) continues to increase with no sign of saturation up to 60 T, although the vortices vastly outnumber pinning centers. We find extremely high Hirr, namely Hirr = 30 T (H||45°) and 24 T (H||c) at 65 K and 58 T (H||45°) and 45 T (H||c) at 50K. The difference in pinning landscape shifts the vortex solid-liquid transition upwards, increasing the vortex region useful for power applications, while keeping the upper critical field, critical temperature and electronic mass anisotropy unchanged. PMID:26853703

  6. Transcriptional Regulation of Zein Gene Expression in Maize through the Additive and Synergistic Action of opaque2, Prolamine-Box Binding Factor, and O2 Heterodimerizing Proteins

    PubMed Central

    Zhang, Zhiyong; Yang, Jun; Wu, Yongrui

    2015-01-01

    Maize (Zea mays) zeins are some of the most abundant cereal seed storage proteins (SSPs). Their abundance influences kernel hardness but compromises its nutritional quality. Transcription factors regulating the expression of zein and other SSP genes in cereals are endosperm-specific and homologs of maize opaque2 (O2) and prolamine-box binding factor (PBF). This study demonstrates that the ubiquitously expressed transcription factors, O2 heterodimerizing proteins (OHPs), specifically regulate 27-kD γ-zein gene expression (through binding to an O2-like box in its promoter) and interact with PBF. The zein content of double mutants OhpRNAi;o2 and PbfRNAi;o2 and the triple mutant PbfRNAi;OhpRNAi;o2 is reduced by 83, 89, and 90%, respectively, compared with the wild type. The triple mutant developed the smallest zein protein bodies, which were merely one-tenth the wild type’s size. Total protein levels in these mutants were maintained in a relatively constant range through proteome rebalancing. These data show that OHPs, O2, and PBF are master regulators of zein storage protein synthesis, acting in an additive and synergistic mode. The differential expression patterns of OHP and O2 genes may cause the slight differences in the timing of 27-kD γ-zein and 22-kD α-zein accumulation during protein body formation. PMID:25901087

  7. Progression from high insulin resistance to type 2 diabetes does not entail additional visceral adipose tissue inflammation.

    PubMed

    Barbarroja, Nuria; Lopez-Pedrera, Chary; Garrido-Sanchez, Lourdes; Mayas, Maria Dolores; Oliva-Olivera, Wilfredo; Bernal-Lopez, Maria Rosa; El Bekay, Rajaa; Tinahones, Francisco Jose

    2012-01-01

    Obesity is associated with a low-grade chronic inflammation state. As a consequence, adipose tissue expresses pro-inflammatory cytokines that propagate inflammatory responses systemically elsewhere, promoting whole-body insulin resistance and consequential islet β-cell exhaustation. Thus, insulin resistance is considered the early stage of type 2 diabetes. However, there is evidence of obese individuals that never develop diabetes indicating that the mechanisms governing the association between the increase of inflammatory factors and type 2 diabetes are much more complex and deserve further investigation. We studied for the first time the differences in insulin signalling and inflammatory pathways in blood and visceral adipose tissue (VAT) of 20 lean healthy donors and 40 equal morbidly obese (MO) patients classified in high insulin resistance (high IR) degree and diabetes state. We studied the changes in proinflammatory markers and lipid content from serum; macrophage infiltration, mRNA expression of inflammatory cytokines and transcription factors, activation of kinases involved in inflammation and expression of insulin signalling molecules in VAT. VAT comparison of these experimental groups revealed that type 2 diabetic-MO subjects exhibit the same pro-inflammatory profile than the high IR-MO patients, characterized by elevated levels of IL-1β, IL-6, TNFα, JNK1/2, ERK1/2, STAT3 and NFκB. Our work rules out the assumption that the inflammation should be increased in obese people with type 2 diabetes compared to high IR obese. These findings indicate that some mechanisms, other than systemic and VAT inflammation must be involved in the development of type 2 diabetes in obesity.

  8. Progression from High Insulin Resistance to Type 2 Diabetes Does Not Entail Additional Visceral Adipose Tissue Inflammation

    PubMed Central

    Barbarroja, Nuria; Lopez-Pedrera, Chary; Garrido-Sanchez, Lourdes; Mayas, Maria Dolores; Oliva-Olivera, Wilfredo; Bernal-Lopez, Maria Rosa; El Bekay, Rajaa; Tinahones, Francisco Jose

    2012-01-01

    Obesity is associated with a low-grade chronic inflammation state. As a consequence, adipose tissue expresses pro-inflammatory cytokines that propagate inflammatory responses systemically elsewhere, promoting whole-body insulin resistance and consequential islet β-cell exhaustation. Thus, insulin resistance is considered the early stage of type 2 diabetes. However, there is evidence of obese individuals that never develop diabetes indicating that the mechanisms governing the association between the increase of inflammatory factors and type 2 diabetes are much more complex and deserve further investigation. We studied for the first time the differences in insulin signalling and inflammatory pathways in blood and visceral adipose tissue (VAT) of 20 lean healthy donors and 40 equal morbidly obese (MO) patients classified in high insulin resistance (high IR) degree and diabetes state. We studied the changes in proinflammatory markers and lipid content from serum; macrophage infiltration, mRNA expression of inflammatory cytokines and transcription factors, activation of kinases involved in inflammation and expression of insulin signalling molecules in VAT. VAT comparison of these experimental groups revealed that type 2 diabetic-MO subjects exhibit the same pro-inflammatory profile than the high IR-MO patients, characterized by elevated levels of IL-1β, IL-6, TNFα, JNK1/2, ERK1/2, STAT3 and NFκB. Our work rules out the assumption that the inflammation should be increased in obese people with type 2 diabetes compared to high IR obese. These findings indicate that some mechanisms, other than systemic and VAT inflammation must be involved in the development of type 2 diabetes in obesity. PMID:23110196

  9. Multiple ribosomal proteins are expressed at high levels in developing zebrafish endoderm and are required for normal exocrine pancreas development.

    PubMed

    Provost, Elayne; Weier, Christopher A; Leach, Steven D

    2013-06-01

    Ribosomal protein L (rpl) genes are essential for assembly of the 60S subunit of the eukaryotic ribosome and may also carry out additional extra-ribosomal functions. We have identified a common expression pattern for rpl genes in developing zebrafish larvae. After initially widespread expression in early embryos, the expression of multiple rpl genes becomes increasingly restricted to the endoderm. With respect to the pancreas, rpl genes are highly expressed in ptf1a-expressing pancreatic progenitors at 48 hpf, suggesting possible functional roles in pancreatic morphogenesis and/or differentiation. Utilizing two available mutant lines, rpl23a(hi2582) and rpl6(hi3655b), we found that ptf1a-expressing pancreatic progenitors fail to properly expand in embryos homozygous for either of these genes. In addition to these durable homozygous phenotypes, we also demonstrated recoverable delays in ptf1a-expressing pancreatic progenitor expansion in rpl23a(hi2582) and rpl6(hi3655b) heterozygotes. Disruptions in ribosome assembly are generally understood to initiate a p53-dependent cellular stress response. However, concomitant p53 knockdown was unable to rescue normal pancreatic progenitor expansion in either rpl23a(hi2582) or rpl6(hi3655b) mutant embryos, suggesting required and p53-independent roles for rpl23a and rpl6 in pancreas development.

  10. High Elmo1 expression aggravates and low Elmo1 expression prevents diabetic nephropathy

    PubMed Central

    Hathaway, Catherine K.; Chang, Albert S.; Grant, Ruriko; Kim, Hyung-Suk; Madden, Victoria J.; Bagnell, C. Robert; Jennette, J. Charles; Smithies, Oliver; Kakoki, Masao

    2016-01-01

    Human genome-wide association studies have demonstrated that polymorphisms in the engulfment and cell motility protein 1 gene (ELMO1) are strongly associated with susceptibility to diabetic nephropathy. However, proof of causation is lacking. To test whether modest changes in its expression alter the severity of the renal phenotype in diabetic mice, we have generated mice that are type 1 diabetic because they have the Ins2Akita gene, and also have genetically graded expression of Elmo1 in all tissues ranging in five steps from ∼30% to ∼200% normal. We here show that the Elmo1 hypermorphs have albuminuria, glomerulosclerosis, and changes in the ultrastructure of the glomerular basement membrane that increase in severity in parallel with the expression of Elmo 1. Progressive changes in renal mRNA expression of transforming growth factor β1 (TGFβ1), endothelin-1, and NAD(P)H oxidase 4 also occur in parallel with Elmo1, as do the plasma levels of cystatin C, lipid peroxides, and TGFβ1, and erythrocyte levels of reduced glutathione. In contrast, Akita type 1 diabetic mice with below-normal Elmo1 expression have reduced expression of these various factors and less severe diabetic complications. Remarkably, the reduced Elmo1 expression in the 30% hypomorphs almost abolishes the pathological features of diabetic nephropathy, although it does not affect the hyperglycemia caused by the Akita mutation. Thus, ELMO1 plays an important role in the development of type 1 diabetic nephropathy, and its inhibition could be a promising option for slowing or preventing progression of the condition to end-stage renal disease. PMID:26858454

  11. Recombinant cells that highly express chromosomally-integrated heterologous genes

    DOEpatents

    Ingram, Lonnie O.; Ohta, Kazuyoshi; Wood, Brent E.

    1998-01-01

    Recombinant host cells are obtained that comprise (A) a heterologous, polypeptide-encoding polynucleotide segment, stably integrated into a chromosome, which is under transcriptional control of an endogenous promoter and (B) a mutation that effects increased expression of the heterologous segment, resulting in enhanced production by the host cells of each polypeptide encoded by that segment, relative to production of each polypeptide by the host cells in the absence of the mutation. The increased expression thus achieved is retained in the absence of conditions that select for cells displaying such increased expression. When the integrated segment comprises, for example, ethanol-production genes from an efficient ethanol producer like Zymomonas mobilis, recombinant Escherichia coli and other enteric bacterial cells within the present invention are capable of converting a wide range of biomass-derived sugars efficiently to ethanol.

  12. Recombinant cells that highly express chromosomally-integrated heterologous genes

    DOEpatents

    Ingram, Lonnie O.; Ohta, Kazuyoshi; Wood, Brent E.

    2000-08-22

    Recombinant host cells are obtained that comprise (A) a heterologous, polypeptide-encoding polynucleotide segment, stably integrated into a chromosome, which is under transcriptional control of an endogenous promoter and (B) a mutation that effects increased expression of the heterologous segment, resulting in enhanced production by the host cells of each polypeptide encoded by that segment, relative to production of each polypeptide by the host cells in the absence of the mutation. The increased expression thus achieved is retained in the absence of conditions that select for cells displaying such increased expression. When the integrated segment comprises, for example, ethanol-production genes from an efficient ethanol producer like Zymomonas mobilis, recombinant Escherichia coli and other enteric bacterial cells within the present invention are capable of converting a wide range of biomass-derived sugars efficiently to ethanol.

  13. Recombinant cells that highly express chromosomally-integrated heterologous genes

    DOEpatents

    Ingram, L.O.; Ohta, Kazuyoshi; Wood, B.E.

    1998-10-13

    Recombinant host cells are obtained that comprise (A) a heterologous, polypeptide-encoding polynucleotide segment, stably integrated into a chromosome, which is under transcriptional control of an endogenous promoter and (B) a mutation that effects increased expression of the heterologous segment, resulting in enhanced production by the host cells of each polypeptide encoded by that segment, relative to production of each polypeptide by the host cells in the absence of the mutation. The increased expression thus achieved is retained in the absence of conditions that select for cells displaying such increased expression. When the integrated segment comprises, for example, ethanol-production genes from an efficient ethanol producer like Zymomonas mobilis, recombinant Escherichia coli and other enteric bacterial cells within the present invention are capable of converting a wide range of biomass-derived sugars efficiently to ethanol. 13 figs.

  14. Recombinant cells that highly express chromosomally-integrated heterologous gene

    DOEpatents

    Ingram, Lonnie O.; Ohta, Kazuyoshi; Wood, Brent E.

    2007-03-20

    Recombinant host cells are obtained that comprise (A) a heterologous, polypeptide-encoding polynucleotide segment, stably integrated into a chromosome, which is under transcriptional control of an endogenous promoter and (B) a mutation that effects increased expression of the heterologous segment, resulting in enhanced production by the host cells of each polypeptide encoded by that segment, relative to production of each polypeptide by the host cells in the absence of the mutation. The increased expression thus achieved is retained in the absence of conditions that select for cells displaying such increased expression. When the integrated segment comprises, for example, ethanol-production genes from an efficient ethanol producer like Zymomonas mobilis, recombinant Escherichia coli and other enteric bacterial cells within the present invention are capable of converting a wide range of biomass-derived sugars efficiently to ethanol.

  15. The effect of a chemical additive on the fermentation and aerobic stability of high-moisture corn.

    PubMed

    Da Silva, T C; Smith, M L; Barnard, A M; Kung, L

    2015-12-01

    The objective of this experiment was to evaluate the effect of a chemical additive on the fermentation and aerobic stability of high-moisture corn (HMC). Ground HMC (~63% dry matter) was untreated, or treated with an additive containing sodium benzoate, potassium sorbate, and sodium nitrite as active ingredients, at 0, 2, 3, or 4 L/t of fresh matter. Laboratory silos (7.5 L) were prepared and ensiled for 21 and 90d (4 silos/treatment per d of ensiling). Small bag silos were prepared for untreated HMC and HMC treated with 4 L/t of the additive and analyzed for nitrate-N and nitrite-N after 0, 3, and 7d of ensiling. The concentration of nitrate-N was similar between these 2 treatments and was below levels considered problematic for ruminants. Nitrite-N was greater in HMC treated with the high level of additive but was also very low for both treatments. Numbers of yeasts were similar among treatments in fresh HMC and decreased substantially after ensiling. Numbers of yeasts were similar among treatments after 21d of ensiling but after 90d they were lower in treated versus untreated HMC. Concentrations of organic acids (lactic, acetic, and propionic) and pH were not different among treatments at any time of ensiling. In contrast, treatment with the additive markedly decreased the concentration of ethanol in HMC after 21 and 90d when compared with untreated HMC. Treatment with all levels of the additive markedly improved the aerobic stability and improved the recovery of dry matter compared with untreated HMC. Overall, our findings suggest that the chemical additive used in this study has the potential to improve the fermentation and aerobic stability of HMC after a relatively short period (21d) and after a moderate length (90d) of ensiling. PMID:26454298

  16. Nontargeted Screening Method for Illegal Additives Based on Ultrahigh-Performance Liquid Chromatography-High-Resolution Mass Spectrometry.

    PubMed

    Fu, Yanqing; Zhou, Zhihui; Kong, Hongwei; Lu, Xin; Zhao, Xinjie; Chen, Yihui; Chen, Jia; Wu, Zeming; Xu, Zhiliang; Zhao, Chunxia; Xu, Guowang

    2016-09-01

    Identification of illegal additives in complex matrixes is important in the food safety field. In this study a nontargeted screening strategy was developed to find illegal additives based on ultrahigh-performance liquid chromatography-high-resolution mass spectrometry (UHPLC-HRMS). First, an analytical method for possible illegal additives in complex matrixes was established including fast sample pretreatment, accurate UHPLC separation, and HRMS detection. Second, efficient data processing and differential analysis workflow were suggested and applied to find potential risk compounds. Third, structure elucidation of risk compounds was performed by (1) searching online databases [Metlin and the Human Metabolome Database (HMDB)] and an in-house database which was established at the above-defined conditions of UHPLC-HRMS analysis and contains information on retention time, mass spectra (MS), and tandem mass spectra (MS/MS) of 475 illegal additives, (2) analyzing fragment ions, and (3) referring to fragmentation rules. Fish was taken as an example to show the usefulness of the nontargeted screening strategy, and six additives were found in suspected fish samples. Quantitative analysis was further carried out to determine the contents of these compounds. The satisfactory application of this strategy in fish samples means that it can also be used in the screening of illegal additives in other kinds of food samples.

  17. Nontargeted Screening Method for Illegal Additives Based on Ultrahigh-Performance Liquid Chromatography-High-Resolution Mass Spectrometry.

    PubMed

    Fu, Yanqing; Zhou, Zhihui; Kong, Hongwei; Lu, Xin; Zhao, Xinjie; Chen, Yihui; Chen, Jia; Wu, Zeming; Xu, Zhiliang; Zhao, Chunxia; Xu, Guowang

    2016-09-01

    Identification of illegal additives in complex matrixes is important in the food safety field. In this study a nontargeted screening strategy was developed to find illegal additives based on ultrahigh-performance liquid chromatography-high-resolution mass spectrometry (UHPLC-HRMS). First, an analytical method for possible illegal additives in complex matrixes was established including fast sample pretreatment, accurate UHPLC separation, and HRMS detection. Second, efficient data processing and differential analysis workflow were suggested and applied to find potential risk compounds. Third, structure elucidation of risk compounds was performed by (1) searching online databases [Metlin and the Human Metabolome Database (HMDB)] and an in-house database which was established at the above-defined conditions of UHPLC-HRMS analysis and contains information on retention time, mass spectra (MS), and tandem mass spectra (MS/MS) of 475 illegal additives, (2) analyzing fragment ions, and (3) referring to fragmentation rules. Fish was taken as an example to show the usefulness of the nontargeted screening strategy, and six additives were found in suspected fish samples. Quantitative analysis was further carried out to determine the contents of these compounds. The satisfactory application of this strategy in fish samples means that it can also be used in the screening of illegal additives in other kinds of food samples. PMID:27480407

  18. HMGA2, the Architectural Transcription Factor High Mobility Group, Is Expressed in the Developing and Mature Mouse Cochlea

    PubMed Central

    Smeti, Ibtihel; Watabe, Isabelle; Savary, Etienne; Fontbonne, Arnaud; Zine, Azel

    2014-01-01

    Hmga2 protein belongs to the non-histone chromosomal high-mobility group (HMG) protein family. HMG proteins have been shown to function as architectural transcription regulators, facilitating enhanceosome formation on a variety of mammalian promoters. Hmga2 are expressed at high levels in embryonic and transformed cells. Terminally differentiated cells, however, have been reported to express only minimal, if any, Hmga2. Our previous affymetrix array data showed that Hmga2 is expressed in the developing and adult mammalian cochleas. However, the spatio-temporal expression pattern of Hmga2 in the murine cochlea remained unknown. In this study, we report the expression of Hmga2 in developing and adult cochleas using immunohistochemistry and quantitative real time PCR analysis. Immunolabeling of Hmga2 in the embryonic, postnatal, and mature cochleas showed broad Hmga2 expression in embryonic cochlea (E14.5) at the level of the developing organ of Corti in differentiating hair cells, supporting cells, in addition to immature cells in the GER and LER areas. By postnatal stage (P0–P3), Hmga2 is predominantly expressed in the hair and supporting cells, in addition to cells in the LER area. By P12, Hmga2 immunolabeling is confined to the hair cells and supporting cells. In the adult ear, Hmga2 expression is maintained in the hair and supporting cell subtypes (i.e. Deiters’ cells, Hensen cells, pillar cells, inner phalangeal and border cells) in the cochlear epithelium. Using quantitative real time PCR, we found a decrease in transcript level for Hmga2 comparable to other known inner ear developmental genes (Sox2, Atoh1, Jagged1 and Hes5) in the cochlear epithelium of the adult relative to postnatal ears. These data provide for the first time the tissue-specific expression and transcription level of Hmga2 during inner ear development and suggest its potential dual role in early differentiation and maintenance of both hair and supporting cell phenotypes. PMID:24551154

  19. A new highly effective anticysticercosis vaccine expressed in transgenic papaya.

    PubMed

    Hernández, Marisela; Cabrera-Ponce, José Luis; Fragoso, Gladis; López-Casillas, Fernando; Guevara-García, Arturo; Rosas, Gabriela; León-Ramírez, Claudia; Juárez, Patricia; Sánchez-García, Guadalupe; Cervantes, Jaquelynne; Acero, Gonzalo; Toledo, Andrea; Cruz, Carmen; Bojalil, Rafael; Herrera-Estrella, Luis; Sciutto, Edda

    2007-05-22

    The use of transgenic plants as new antigen-delivery systems for subunit vaccines has been increasingly explored. We herein report progress toward a papaya-based vaccine against cysticercosis. Synthetic peptides (KETc1, KETc12, KETc7) were successfully expressed in 19 different transgenic papaya clones and found to be immunogenic. Complete protection against cysticercosis was induced with the soluble extract of the clones that expressed the higher levels of transcripts in up to 90% of the immunized mice. This study represents a key step towards the development of a more effective, sustainable and affordable oral subunit vaccine against human and pig cysticercosis.

  20. [High Throughput Screening Analysis of Preservatives and Sweeteners in Carbonated Beverages Based on Improved Standard Addition Method].

    PubMed

    Wang, Su-fang; Liu, Yun; Gong, Li-hua; Dong, Chun-hong; Fu, De-xue; Wang, Guo-qing

    2016-02-01

    Simulated water samples of 3 kinds of preservatives and 4 kinds of sweeteners were formulated by using orthogonal design. Kernel independent component analysis (KICA) was used to process the UV spectra of the simulated water samples and the beverages added different amounts of the additive standards, then the independent components (ICs), i. e. the UV spectral profiles of the additives, and the ICs' coefficient matrices were used to establish UV-KICA-SVR prediction model of the simulated preservatives and sweeteners solutions using support vector regression (SVR) analysis. The standards added beverages samples were obtained by adding different amounts level of additives in carbonated beverages, their UV spectra were processed by KICA, then IC information represented to the additives and other sample matrix were obtained, and the sample background can be deducted by removing the corresponding IC, other ICs' coefficient matrices were used to estimate the amounts of the additives in the standard added beverage samples based on the UV-KICA-SVR model, while the intercept of linear regression equation of predicted amounts and the added amounts in the standard added samples is the additive content in the raw beverage sample. By utilization of chemometric "blind source separation" method for extracting IC information of the tested additives in the beverage and other sample matrix, and using SVR regression modeling to improve the traditional standard addition method, a new method was proposed for the screening of the preservatives and sweeteners in carbonated beverages. The proposed UV-KICA-SVR method can be used to determine 3 kinds of preservatives and 4 kinds of sweetener in the carbonate beverages with the limit of detection (LOD) are located with the range 0.2-1.0 mg · L⁻¹, which are comparable to that of the traditional high performance liquid chromatographic (HPLC) method. PMID:27209754

  1. [High Throughput Screening Analysis of Preservatives and Sweeteners in Carbonated Beverages Based on Improved Standard Addition Method].

    PubMed

    Wang, Su-fang; Liu, Yun; Gong, Li-hua; Dong, Chun-hong; Fu, De-xue; Wang, Guo-qing

    2016-02-01

    Simulated water samples of 3 kinds of preservatives and 4 kinds of sweeteners were formulated by using orthogonal design. Kernel independent component analysis (KICA) was used to process the UV spectra of the simulated water samples and the beverages added different amounts of the additive standards, then the independent components (ICs), i. e. the UV spectral profiles of the additives, and the ICs' coefficient matrices were used to establish UV-KICA-SVR prediction model of the simulated preservatives and sweeteners solutions using support vector regression (SVR) analysis. The standards added beverages samples were obtained by adding different amounts level of additives in carbonated beverages, their UV spectra were processed by KICA, then IC information represented to the additives and other sample matrix were obtained, and the sample background can be deducted by removing the corresponding IC, other ICs' coefficient matrices were used to estimate the amounts of the additives in the standard added beverage samples based on the UV-KICA-SVR model, while the intercept of linear regression equation of predicted amounts and the added amounts in the standard added samples is the additive content in the raw beverage sample. By utilization of chemometric "blind source separation" method for extracting IC information of the tested additives in the beverage and other sample matrix, and using SVR regression modeling to improve the traditional standard addition method, a new method was proposed for the screening of the preservatives and sweeteners in carbonated beverages. The proposed UV-KICA-SVR method can be used to determine 3 kinds of preservatives and 4 kinds of sweetener in the carbonate beverages with the limit of detection (LOD) are located with the range 0.2-1.0 mg · L⁻¹, which are comparable to that of the traditional high performance liquid chromatographic (HPLC) method.

  2. Gene expression profiles in liver of pigs with extreme high and low levels of androstenone

    PubMed Central

    Moe, Maren; Lien, Sigbjørn; Bendixen, Christian; Hedegaard, Jakob; Hornshøj, Henrik; Berget, Ingunn; Meuwissen, Theo HE; Grindflek, Eli

    2008-01-01

    Background Boar taint is the unpleasant odour and flavour of the meat of uncastrated male pigs that is primarily caused by high levels of androstenone and skatole in adipose tissue. Androstenone is a steroid and its levels are mainly genetically determined. Studies on androstenone metabolism have, however, focused on a limited number of genes. Identification of additional genes influencing levels of androstenone may facilitate implementation of marker assisted breeding practices. In this study, microarrays were used to identify differentially expressed genes and pathways related to androstenone metabolism in the liver from boars with extreme levels of androstenone in adipose tissue. Results Liver tissue samples from 58 boars of the two breeds Duroc and Norwegian Landrace, 29 with extreme high and 29 with extreme low levels of androstenone, were selected from more than 2500 individuals. The samples were hybridised to porcine cDNA microarrays and the 1% most significant differentially expressed genes were considered significant. Among the differentially expressed genes were metabolic phase I related genes belonging to the cytochrome P450 family and the flavin-containing monooxygenase FMO1. Additionally, phase II conjugation genes including UDP-glucuronosyltransferases UGT1A5, UGT2A1 and UGT2B15, sulfotransferase STE, N-acetyltransferase NAT12 and glutathione S-transferase were identified. Phase I and phase II metabolic reactions increase the water solubility of steroids and play a key role in their elimination. Differential expression was also found for genes encoding 17beta-hydroxysteroid dehydrogenases (HSD17B2, HSD17B4, HSD17B11 and HSD17B13) and plasma proteins alpha-1-acid glycoprotein (AGP) and orosomucoid (ORM1). 17beta-hydroxysteroid dehydrogenases and plasma proteins regulate the availability of steroids by controlling the amount of active steroids accessible to receptors and available for metabolism. Differences in the expression of FMO1, NAT12, HSD17B2 and

  3. Development and molecular characterization of wheat--Aegilops kotschyi addition and substitution lines with high grain protein, iron, and zinc.

    PubMed

    Rawat, Nidhi; Neelam, Kumari; Tiwari, Vijay K; Randhawa, Gursharn S; Friebe, Bernd; Gill, Bikram S; Dhaliwal, Harcharan S

    2011-11-01

    Over two billion people, depending largely on staple foods, suffer from deficiencies in protein and some micronutrients such as iron and zinc. Among various approaches to overcome protein and micronutrient deficiencies, biofortification through a combination of conventional and molecular breeding methods is the most feasible, cheapest, and sustainable approach. An interspecific cross was made between the wheat cultivar 'Chinese Spring' and Aegilops kotschyi Boiss. accession 396, which has a threefold higher grain iron and zinc concentrations and about 33% higher protein concentration than wheat cultivars. Recurrent backcrossing and selection for the micronutrient content was performed at each generation. Thirteen derivatives with high grain iron and zinc concentrations and contents, ash and ash micronutrients, and protein were analyzed for alien introgression. Morphological markers, high molecular weight glutenin subunit profiles, anchored wheat microsatellite markers, and GISH showed that addition and substitution of homoeologous groups 1, 2, and 7 chromosomes of Ae. kotschyi possess gene(s) for high grain micronutrients. The addition of 1U/1S had high molecular weight glutenin subunits with higher molecular weight than those of wheat, and the addition of 2S in most of the derivatives also enhanced grain protein content by over 20%. Low grain protein content in a derivative with a 2S-wheat translocation, waxy leaves, and absence of the gdm148 marker strongly suggests that the gene for higher grain protein content on chromosome 2S is orthologous to the grain protein QTL on the short arm of group 2 chromosomes.

  4. High-Dimensional Gene Expression Profiling Studies in High and Low Responders to Primary Smallpox Vaccination

    PubMed Central

    Haralambieva, Iana H.; Oberg, Ann L.; Dhiman, Neelam; Ovsyannikova, Inna G.; Kennedy, Richard B.; Grill, Diane E.; Jacobson, Robert M.; Poland, Gregory A.

    2012-01-01

    Background. The mechanisms underlying smallpox vaccine-induced variations in immune responses are not well understood, but are of considerable interest to a deeper understanding of poxvirus immunity and correlates of protection. Methods. We assessed transcriptional messenger RNA expression changes in 197 recipients of primary smallpox vaccination representing the extremes of humoral and cellular immune responses. Results. The 20 most significant differentially expressed genes include a tumor necrosis factor–receptor superfamily member, an interferon (IFN) gene, a chemokine gene, zinc finger protein genes, nuclear factors, and histones (P ≤ 1.06E−20, q ≤ 2.64E−17). A pathway analysis identified 4 enriched pathways with cytokine production by the T-helper 17 subset of CD4+ T cells being the most significant pathway (P = 3.42E−05). Two pathways (antiviral actions of IFNs, P = 8.95E−05; and IFN-α/β signaling pathway, P = 2.92E−04), integral to innate immunity, were enriched when comparing high with low antibody responders (false discovery rate, < 0.05). Genes related to immune function and transcription (TLR8, P = .0002; DAPP1, P = .0003; LAMP3, P = 9.96E−05; NR4A2, P ≤ .0002; EGR3, P = 4.52E−05), and other genes with a possible impact on immunity (LNPEP, P = 3.72E−05; CAPRIN1, P = .0001; XRN1, P = .0001), were found to be expressed differentially in high versus low antibody responders. Conclusion. We identified novel and known immunity-related genes and pathways that may account for differences in immune response to smallpox vaccination. PMID:22949304

  5. The effect of trace element addition to mono-digestion of grass silage at high organic loading rates.

    PubMed

    Wall, David M; Allen, Eoin; Straccialini, Barbara; O'Kiely, Padraig; Murphy, Jerry D

    2014-11-01

    This study investigated the effect of trace element addition to mono-digestion of grass silage at high organic loading rates. Two continuous reactors were compared. The first mono-digested grass silage whilst the second operated in co-digestion, 80% grass silage with 20% dairy slurry (VS basis). The reactors were run for 65weeks with a further 5weeks taken for trace element supplementation for the mono-digestion of grass silage. The co-digestion reactor reported a higher biomethane efficiency (1.01) than mono-digestion (0.90) at an OLR of 4.0kgVSm(-3)d(-1) prior to addition of trace elements. Addition of cobalt, iron and nickel, led to an increase in the SMY in mono-digestion of grass silage by 12% to 404LCH4kg(-1)VS and attained a biomethane efficiency of 1.01.

  6. High performance flexible double-sided micro-supercapacitors with an organic gel electrolyte containing a redox-active additive.

    PubMed

    Kim, Doyeon; Lee, Geumbee; Kim, Daeil; Yun, Junyeong; Lee, Sang-Soo; Ha, Jeong Sook

    2016-08-25

    In this study, we report the fabrication of a high performance flexible micro-supercapacitor (MSC) with an organic gel electrolyte containing a redox-active additive, referred to as poly(methyl methacrylate)-propylene carbonate-lithium perchlorate-hydroquinone (PMMA-PC-LiClO4-HQ). Hexagonal MSCs fabricated on thin polyethylene terephthalate (PET) films had interdigitated electrodes made of spray-coated multi-walled carbon nanotubes (MWNTs) on Au. The addition of HQ as a redox-active additive enhanced not only the specific capacitance but also the energy density of the MSCs dramatically, which is approximately 35 times higher than that of MSCs without the HQ additive. In addition, both areal capacitance and areal energy density could be doubled by fabrication of double-sided MSCs, where two MSCs are connected in parallel. The double-sided MSCs exhibited stable electrochemical performance during repeated deformation by bending. By dry-transferring the double-sided MSCs based on PMMA-PC-LiClO4-HQ on a deformable polymer substrate, we fabricated a stretchable MSC array, which also retained its electrochemical performance during a uniaxial strain of 40%. Furthermore, a wearable energy storage bracelet made of such an MSC array could operate a μ-LED on the wrist. PMID:27511060

  7. The negative effect of Zr addition on the high temperature strength in alumina-forming austenitic stainless steels

    SciTech Connect

    Moon, Joonoh; Jang, Min-Ho; Kang, Jun-Yun; Lee, Tae-Ho

    2014-01-15

    The effect of a Zr addition on the precipitation behavior and mechanical properties in Nb-containing alumina-forming austenitic (AFA) stainless steels was investigated using tensile tests, scanning electron microscopy (SEM), and scanning transmission electron microscopy (STEM) analysis. The TEM observation showed that a Zr addition led to the formation of a (Nb,Zr)(C,N) complex particle, which coarsened the Nb-rich carbonitride. Tensile tests were performed at an elevated temperature (700 °C), and both the tensile and yield strengths decreased with a Zr addition. This unexpected result of a Zr addition was due to the reduction of the precipitation strengthening by particle coarsening. - Highlights: • The effect of Zr on high temperature strength in AFA steel containing Nb was studied. • Both the tensile and yield strengths of an AFA steel decreased with Zr-addition. • This is due to the reduction of precipitation strengthening by particle coarsening. • Nb(C,N) and (Nb,Zr)(C,N) particles were precipitated in an AFA and Zr-added AFA steel. • The size of (Nb,Zr)(C,N) particle is much bigger than that of Nb(C,N) particle.

  8. Gene expression suggests double-segmental and single-segmental patterning mechanisms during posterior segment addition in the beetle Tribolium castaneum.

    PubMed

    Janssen, Ralf

    2014-01-01

    In the model arthropod Drosophila, all segments are patterned simultaneously in the blastoderm. In most other arthropods, however, posterior segments are added sequentially from a posterior segment addition zone. Posterior addition of single segments likely represents the ancestral mode of arthropod segmentation, although in Drosophila, segments are patterned in pairs by the pair-rule genes. It has been shown that in the new model insect, the beetle Tribolium, a segmentation clock operates that apparently patterns all segments in pairs as well. Here, I report on the expression of the segment polarity gene H15/midline in Tribolium. In the anterior embryo, segmental stripes of H15 appear in pairs, but in the posterior of the embryo stripes appear in a single-segmental periodicity. This implies that either two completely different segmentation-mechanisms may act in the germ band of Tribolium, that the segmentation clock changes its periodicity during development, or that the speed in which posterior segments are patterned changes. In any case, the data suggest the presence of another (or modified), yet undiscovered, mechanism of posterior segment addition in one of the best-understood arthropod models. The finding of a hitherto unrecognized segmentation mechanism in Tribolium may have major implications for the understanding of the origin of segmentation mechanisms, including the origin of pair rule patterning. It also calls for (re)-investigation of posterior segment addition in Tribolium and other previously studied arthropod models.

  9. Combinatorial Screening for Transgenic Yeasts with High Cellulase Activities in Combination with a Tunable Expression System.

    PubMed

    Ito, Yoichiro; Yamanishi, Mamoru; Ikeuchi, Akinori; Imamura, Chie; Matsuyama, Takashi

    2015-01-01

    Combinatorial screening used together with a broad library of gene expression cassettes is expected to produce a powerful tool for the optimization of the simultaneous expression of multiple enzymes. Recently, we proposed a highly tunable protein expression system that utilized multiple genome-integrated target genes to fine-tune enzyme expression in yeast cells. This tunable system included a library of expression cassettes each composed of three gene-expression control elements that in different combinations produced a wide range of protein expression levels. In this study, four gene expression cassettes with graded protein expression levels were applied to the expression of three cellulases: cellobiohydrolase 1, cellobiohydrolase 2, and endoglucanase 2. After combinatorial screening for transgenic yeasts simultaneously secreting these three cellulases, we obtained strains with higher cellulase expressions than a strain harboring three cellulase-expression constructs within one high-performance gene expression cassette. These results show that our method will be of broad use throughout the field of metabolic engineering.

  10. Combinatorial Screening for Transgenic Yeasts with High Cellulase Activities in Combination with a Tunable Expression System.

    PubMed

    Ito, Yoichiro; Yamanishi, Mamoru; Ikeuchi, Akinori; Imamura, Chie; Matsuyama, Takashi

    2015-01-01

    Combinatorial screening used together with a broad library of gene expression cassettes is expected to produce a powerful tool for the optimization of the simultaneous expression of multiple enzymes. Recently, we proposed a highly tunable protein expression system that utilized multiple genome-integrated target genes to fine-tune enzyme expression in yeast cells. This tunable system included a library of expression cassettes each composed of three gene-expression control elements that in different combinations produced a wide range of protein expression levels. In this study, four gene expression cassettes with graded protein expression levels were applied to the expression of three cellulases: cellobiohydrolase 1, cellobiohydrolase 2, and endoglucanase 2. After combinatorial screening for transgenic yeasts simultaneously secreting these three cellulases, we obtained strains with higher cellulase expressions than a strain harboring three cellulase-expression constructs within one high-performance gene expression cassette. These results show that our method will be of broad use throughout the field of metabolic engineering. PMID:26692026

  11. Combinatorial Screening for Transgenic Yeasts with High Cellulase Activities in Combination with a Tunable Expression System

    PubMed Central

    Ito, Yoichiro; Yamanishi, Mamoru; Ikeuchi, Akinori; Imamura, Chie; Matsuyama, Takashi

    2015-01-01

    Combinatorial screening used together with a broad library of gene expression cassettes is expected to produce a powerful tool for the optimization of the simultaneous expression of multiple enzymes. Recently, we proposed a highly tunable protein expression system that utilized multiple genome-integrated target genes to fine-tune enzyme expression in yeast cells. This tunable system included a library of expression cassettes each composed of three gene-expression control elements that in different combinations produced a wide range of protein expression levels. In this study, four gene expression cassettes with graded protein expression levels were applied to the expression of three cellulases: cellobiohydrolase 1, cellobiohydrolase 2, and endoglucanase 2. After combinatorial screening for transgenic yeasts simultaneously secreting these three cellulases, we obtained strains with higher cellulase expressions than a strain harboring three cellulase-expression constructs within one high-performance gene expression cassette. These results show that our method will be of broad use throughout the field of metabolic engineering. PMID:26692026

  12. Phytate addition to soil induces changes in the abundance and expression of Bacillus β-propeller phytase genes in the rhizosphere.

    PubMed

    Jorquera, Milko A; Saavedra, Nicolás; Maruyama, Fumito; Richardson, Alan E; Crowley, David E; del C Catrilaf, Rosa; Henriquez, Evelyn J; de la Luz Mora, María

    2013-02-01

    Phytate-mineralizing rhizobacteria (PMR) perform an essential function for the mineralization of organic phosphorus but little is known about their ecology in soils and rhizosphere. In this study, PCR-based methods were developed for detection and quantification of the Bacillus β-propeller phytase (BPP) gene. Experiments were conducted to monitor the presence and persistence of a phytate-mineralizing strain, Bacillus sp. MQH19, after inoculation of soil microcosms and within the rhizosphere. The occurrence of the BPP gene in natural pasture soils from Chilean Andisols was also examined. The results showed that the Bacillus BPP gene was readily detected in sterile and nonsterile microcosms, and that the quantitative PCR (qPCR) methods could be used to monitor changes in the abundance of the BPP gene over time. Our results also show that the addition of phytate to nonsterile soils induced the expression of the BPP gene in the rhizosphere of ryegrass and the BPP gene was detected in all pasture soils sampled. This study shows that phytate addition soils induced changes in the abundance and expression of Bacillus BPP to genes in the rhizosphere and demonstrates that Bacillus BPP gene is cosmopolitan in pasture soils from Chilean Andisols.

  13. High Expression of MicroRNA-196a Indicates Poor Prognosis in Resected Pancreatic Neuroendocrine Tumor

    PubMed Central

    Lee, Yoon Suk; Kim, Haeryoung; Kim, Hyoung Woo; Lee, Jong-Chan; Paik, Kyu-Hyun; Kang, Jingu; Kim, Jaihwan; Yoon, Yoo-Seok; Han, Ho-Seong; Sohn, Insuk; Cho, Jeonghee; Hwang, Jin-Hyeok

    2015-01-01

    Abstract There is limited data on miRNA expression in pancreatic neuroendocrine tumors (PanNETs). In this study, we aimed to identify miRNAs that could be potential prognostic biomarkers of PanNETs in patients who underwent curative surgery. For miRNA target screening, 2 primary PanNETs and corresponding liver metastases were screened for miRNA expression by the NanoString nCounter analysis. Candidate miRNAs were selected by ≥2-fold difference of expression between metastatic versus primary tumor. For miRNA target validation, quantitative real-time PCR was performed for candidate miRNAs on 37 PanNETs and matched nonneoplastic pancreata, and the miRNA levels were correlated with the clinicopathological features and patient survival data. Eight miRNAs (miRNA-27b, -122, -142–5p, -196a, -223, -590–5p, -630, and -944) were selected as candidate miRNAs. Only miR-196a level was significantly associated with stage, and mitotic count. When PanNETs were stratified into high (n = 10) and low (n = 27) miRNA-196a expression groups, miRNA-196a-high PanNETs were significantly associated with advanced pathologic T stage (50.0% vs 7.4%), N stage (50.0% vs 3.7%), higher mitotic counts (60.0% vs 3.7%), and higher Ki-67-labeling indices (60.0% vs 22.2%). In addition, high miRNA-196a expression was significantly associated with decreased overall survival (P = 0.046) and disease-free survival (P < 0.001) during a median follow-up of 37.9 months with the hazard ratio for recurrence of 16.267 (95% confidence interval = 1.732–153.789; P = 0.015). MiRNA-196a level may be a promising prognostic marker of recurrence in resected PanNETs, although further experimental investigation would be required. PMID:26683934

  14. High expression of green fluorescent protein in Pichia pastoris leads to formation of fluorescent particles.

    PubMed

    Lenassi Zupan, Ana; Trobec, Sonja; Gaberc-Porekar, Vladka; Menart, Viktor

    2004-04-01

    Wild type gene for green fluorescent protein (GFP) was stably integrated into the Pichia pastoris genome and yielded an expression level of over 40% of total cellular protein. The high cytoplasmic concentration of fluorescent (properly folded and processed) GFP caused the formation of fluorescent spherical structures, which could be observed by fluorescence or confocal microscopy after controlled permeabilization of the yeast cells with 0.2% N-lauroyl sarcosine (NLS). Fluorescent GFP particles were also isolated after removal of the cell wall and found to be quite resistant to 0.2% N-lauroyl sarcosine. SDS-PAGE analysis of the isolated fluorescent particles revealed the presence of an 80 kDa protein (alcohol oxidase) and GFP (30%). We conclude that GFP is able to enter spontaneously into the peroxisomes and is inserted into densely packed layers of alcohol oxidase. Consequently, the formation of similar fluorescent particles can also be expected in other organisms when using high-level expression systems. As GFP is widely used in fusion with other proteins as a reporter for protein localization and for many other applications in biotechnology, care must be taken to avoid false interpretations of targeting or trafficking mechanisms inside the cells. In addition, when whole cells or cytoplasmic fractions are used for the quantitative determination of GFP levels, incorrect and misleading values of GFP could be obtained due to the formation of fluorescent particles containing material inside which is not available for fluorescence measurements.

  15. Altered Gene Expression in Mice Selected for High Maternal Aggression

    PubMed Central

    Gammie, Stephen C.; Auger, Anthony P.; Jessen, Heather M.; Vanzo, Rena J.; Awad, Tarif A.; Stevenson, Sharon A.

    2007-01-01

    We previously applied selective breeding on outbred mice to increase maternal aggression (maternal defense). In this study, we compared gene expression within a continuous region of the CNS involved in maternal aggression (hypothalamus and preoptic regions) between lactating selected (S) and non-selected control (C) mice (n = 6 per group). Using microarrays representing over 40,000 genes or expressed sequence tags, two statistical algorithms were used to identify significant differences in gene expression: robust multi array and the probe logarithmic intensity error method. ∼ 200 genes were identified as significant using an intersection from both techniques. A subset of genes were examined for confirmation by real-time PCR. Significant decreases were found in S mice for neurotensin and neuropeptide Y receptor Y2 (both confirmed by PCR). Significant increases were found in S mice for neuronal nitric oxide synthase (confirmed by PCR), the K+ channel subunit, Kcna1 (confirmed by PCR), corticotrophin releasing factor binding protein (just above significance using PCR; p = 0.051), and GABA A receptor subunit 1A (not confirmed by PCR, but similar direction). S mice also exhibited significantly higher levels of the neurotransmitter receptor, adenosine A1 receptor, and the transcription factors, c-Fos, and Egr-1. Interestingly, for 24 genes related to metabolism, all were significantly elevated in S mice, suggesting altered metabolism in these mice. Together, this study provides a list of candidate genes (some previously implicated in maternal aggression and some novel) that may play an important role in the production of this behavior. PMID:16939635

  16. Additive manufacturing of liquid/gas diffusion layers for low-cost and high-efficiency hydrogen production

    DOE PAGES

    Mo, Jingke; Zhang, Feng -Yuan; Dehoff, Ryan R.; Peter, William H.; Toops, Todd J.; Green, Jr., Johney Boyd

    2016-01-14

    The electron beam melting (EBM) additive manufacturing technology was used to fabricate titanium liquid/gas diffusion media with high-corrosion resistances and well-controllable multifunctional parameters, including two-phase transport and excellent electric/thermal conductivities, has been first demonstrated. Their applications in proton exchange membrane eletrolyzer cells have been explored in-situ in a cell and characterized ex-situ with SEM and XRD. Compared with the conventional woven liquid/gas diffusion layers (LGDLs), much better performance with EBM fabricated LGDLs is obtained due to their significant reduction of ohmic loss. The EBM technology components exhibited several distinguished advantages in fabricating gas diffusion layer: well-controllable pore morphology and structure,more » rapid prototyping, fast manufacturing, highly customizing and economic. In addition, by taking advantage of additive manufacturing, it possible to fabricate complicated three-dimensional designs of virtually any shape from a digital model into one single solid object faster, cheaper and easier, especially for titanium. More importantly, this development will provide LGDLs with control of pore size, pore shape, pore distribution, and therefore porosity and permeability, which will be very valuable to develop modeling and to validate simulations of electrolyzers with optimal and repeatable performance. Further, it will lead to a manufacturing solution to greatly simplify the PEMEC/fuel cell components and to couple the LGDLs with other parts, since they can be easily integrated together with this advanced manufacturing process« less

  17. High-level expression of Proteinase K from Tritirachium album Limber in Pichia pastoris using multi-copy expression strains.

    PubMed

    Yang, Hu; Zhai, Chao; Yu, Xianhong; Li, Zhezhe; Tang, Wei; Liu, Yunyun; Ma, Xiaojian; Zhong, Xing; Li, Guolong; Wu, Di; Ma, Lixin

    2016-06-01

    Proteinase K is widely used in scientific research and industries. This report was aimed to achieve high-level expression of proteinase K using Pichia pastoris GS115 as the host strain. The coding sequence of a variant of proteinase K that has higher activity than the wild type protein was chosen and optimized based on the codon usage preference of P. pastoris. The novel open reading frame was synthesized and a series of multi-copy expression vectors were constructed based on the pHBM905BDM plasmid, allowing for the tandem integration of multiple copies of the target gene into the genome of P. pastoris with a single recombination. These strains were used to study the correlation between the gene copy number and the expression level of proteinase K. The results of quantitative polymerase chain reaction (qPCR) indicated that the tandem expression cassettes were integrated into the host genome stably. Meanwhile, the results of qPCR and enzyme activity assays indicated that the mRNA and protein expression levels of the target gene increased as the gene copy number increased. Moreover, the effect of gene dosage on the expression level of the recombinant protein was more obvious using high-density fermentation. The maximum expression level and enzyme activity of proteinase K, which were obtained from the recombinant yeast strain bearing 5 copies of the target gene after an 84-h induction, were approximately 8.069 mg/mL and 108,295 U/mL, respectively. The recombinant proteinase was purified and characterized. The optimum pH and temperature for the activity of this protease were approximately pH 11 and 55 °C, respectively. PMID:26892536

  18. High-level expression of Proteinase K from Tritirachium album Limber in Pichia pastoris using multi-copy expression strains.

    PubMed

    Yang, Hu; Zhai, Chao; Yu, Xianhong; Li, Zhezhe; Tang, Wei; Liu, Yunyun; Ma, Xiaojian; Zhong, Xing; Li, Guolong; Wu, Di; Ma, Lixin

    2016-06-01

    Proteinase K is widely used in scientific research and industries. This report was aimed to achieve high-level expression of proteinase K using Pichia pastoris GS115 as the host strain. The coding sequence of a variant of proteinase K that has higher activity than the wild type protein was chosen and optimized based on the codon usage preference of P. pastoris. The novel open reading frame was synthesized and a series of multi-copy expression vectors were constructed based on the pHBM905BDM plasmid, allowing for the tandem integration of multiple copies of the target gene into the genome of P. pastoris with a single recombination. These strains were used to study the correlation between the gene copy number and the expression level of proteinase K. The results of quantitative polymerase chain reaction (qPCR) indicated that the tandem expression cassettes were integrated into the host genome stably. Meanwhile, the results of qPCR and enzyme activity assays indicated that the mRNA and protein expression levels of the target gene increased as the gene copy number increased. Moreover, the effect of gene dosage on the expression level of the recombinant protein was more obvious using high-density fermentation. The maximum expression level and enzyme activity of proteinase K, which were obtained from the recombinant yeast strain bearing 5 copies of the target gene after an 84-h induction, were approximately 8.069 mg/mL and 108,295 U/mL, respectively. The recombinant proteinase was purified and characterized. The optimum pH and temperature for the activity of this protease were approximately pH 11 and 55 °C, respectively.

  19. High Expression of Water-Soluble Recombinant Antigenic Domains of Toxoplasma gondii Secretory Organelles

    PubMed Central

    Yang, Zhaoshou; Ahn, Hye-Jin

    2014-01-01

    Recombinant antigenic proteins of Toxoplasma gondii are alternative source of antigens which are easily obtainable for serodiagnosis of toxoplasmosis. In this study, highly antigenic secretory organellar proteins, dense granular GRA2 and GRA3, rhoptrial ROP2, and micronemal MIC2, were analyzed by bioinformatics approach to express as water-soluble forms of antigenic domains. The transmembrane region and disorder tendency of 4 secretory proteins were predicted to clone the genes into pGEX-4T-1 vector. Recombinant plasmids were transformed into BL21 (DE3) pLysS E. coli, and GST fusion proteins were expressed with IPTG. As a result, GST fusion proteins with GRA225-105, GRA339-138, ROP2324-561, and MIC21-284 domains had respectively higher value of IgG avidity. The rGST-GRA225-105 and rGST-GRA339-138 were soluble, while rGST-ROP2324-561 and rGST-MIC21-284 were not. GRA231-71, intrinsically unstructured domain (IUD) of GRA2, was used as a linker to enhance the solubility. The rGST-GRA231-71-ROP2324-561, a chimeric protein, appeared to be soluble. Moreover, rGST-GRA231-71-MIC21-284 was also soluble and had higher IgG avidity comparing to rGST-MIC21-284. These 4 highly expressed and water-soluble recombinant antigenic proteins may be promising candidates to improve the serodiagnosis of toxoplasmosis in addition to the major surface antigen of SAG1. PMID:25246715

  20. Parallel Gene Expression Differences between Low and High Latitude Populations of Drosophila melanogaster and D. simulans.

    PubMed

    Zhao, Li; Wit, Janneke; Svetec, Nicolas; Begun, David J

    2015-05-01

    Gene expression variation within species is relatively common, however, the role of natural selection in the maintenance of this variation is poorly understood. Here we investigate low and high latitude populations of Drosophila melanogaster and its sister species, D. simulans, to determine whether the two species show similar patterns of population differentiation, consistent with a role for spatially varying selection in maintaining gene expression variation. We compared at two temperatures the whole male transcriptome of D. melanogaster and D. simulans sampled from Panama City (Panama) and Maine (USA). We observed a significant excess of genes exhibiting differential expression in both species, consistent with parallel adaptation to heterogeneous environments. Moreover, the majority of genes showing parallel expression differentiation showed the same direction of differential expression in the two species and the magnitudes of expression differences between high and low latitude populations were correlated across species, further bolstering the conclusion that parallelism for expression phenotypes results from spatially varying selection. However, the species also exhibited important differences in expression phenotypes. For example, the genomic extent of genotype × environment interaction was much more common in D. melanogaster. Highly differentiated SNPs between low and high latitudes were enriched in the 3' UTRs and CDS of the geographically differently expressed genes in both species, consistent with an important role for cis-acting variants in driving local adaptation for expression-related phenotypes.

  1. Parallel Gene Expression Differences between Low and High Latitude Populations of Drosophila melanogaster and D. simulans

    PubMed Central

    Zhao, Li; Wit, Janneke; Svetec, Nicolas; Begun, David J.

    2015-01-01

    Gene expression variation within species is relatively common, however, the role of natural selection in the maintenance of this variation is poorly understood. Here we investigate low and high latitude populations of Drosophila melanogaster and its sister species, D. simulans, to determine whether the two species show similar patterns of population differentiation, consistent with a role for spatially varying selection in maintaining gene expression variation. We compared at two temperatures the whole male transcriptome of D. melanogaster and D. simulans sampled from Panama City (Panama) and Maine (USA). We observed a significant excess of genes exhibiting differential expression in both species, consistent with parallel adaptation to heterogeneous environments. Moreover, the majority of genes showing parallel expression differentiation showed the same direction of differential expression in the two species and the magnitudes of expression differences between high and low latitude populations were correlated across species, further bolstering the conclusion that parallelism for expression phenotypes results from spatially varying selection. However, the species also exhibited important differences in expression phenotypes. For example, the genomic extent of genotype × environment interaction was much more common in D. melanogaster. Highly differentiated SNPs between low and high latitudes were enriched in the 3’ UTRs and CDS of the geographically differently expressed genes in both species, consistent with an important role for cis-acting variants in driving local adaptation for expression-related phenotypes. PMID:25950438

  2. Rapid plant species loss at high rates and at low frequency of N addition in temperate steppe.

    PubMed

    Zhang, Yunhai; Lü, Xiaotao; Isbell, Forest; Stevens, Carly; Han, Xu; He, Nianpeng; Zhang, Guangming; Yu, Qiang; Huang, Jianhui; Han, Xingguo

    2014-11-01

    Humans are both intentionally (fertilization) and unintentionally (atmospheric nutrient deposition) adding nutrients worldwide. Increasing availability of biologically reactive nitrogen (N) is one of the major drivers of plant species loss. It remains unclear, however, whether plant diversity will be equally reduced by inputs of reactive N coming from either small and frequent N deposition events or large and infrequent N fertilization events. By independently manipulating the rate and frequency of reactive N inputs, our study teases apart these potentially contrasting effects. Plant species richness decreased more quickly at high rates and at low frequency of N addition, which suggests that previous fertilization studies have likely over-estimated the effects of N deposition on plant species loss. N-induced species loss resulted from both acidification and ammonium toxicity. Further study of small and frequent N additions will be necessary to project future rates of plant species loss under increasing aerial N deposition.

  3. New Vectors for Chromosomal Integration Enable High-Level Constitutive or Inducible Magnetosome Expression of Fusion Proteins in Magnetospirillum gryphiswaldense

    PubMed Central

    Borg, Sarah; Hofmann, Julia; Pollithy, Anna; Lang, Claus

    2014-01-01

    The alphaproteobacterium Magnetospirillum gryphiswaldense biomineralizes magnetosomes, which consist of monocrystalline magnetite cores enveloped by a phospholipid bilayer containing specific proteins. Magnetosomes represent magnetic nanoparticles with unprecedented magnetic and physicochemical characteristics. These make them potentially useful in a number of biotechnological and biomedical applications. Further functionalization can be achieved by expression of foreign proteins via genetic fusion to magnetosome anchor peptides. However, the available genetic tool set for strong and controlled protein expression in magnetotactic bacteria is very limited. Here, we describe versatile vectors for either inducible or high-level constitutive expression of proteins in M. gryphiswaldense. The combination of an engineered native PmamDC promoter with a codon-optimized egfp gene (Mag-egfp) resulted in an 8-fold increase in constitutive expression and in brighter fluorescence. We further demonstrate that the widely used Ptet promoter is functional and tunable in M. gryphiswaldense. Stable and uniform expression of the EGFP and β-glucuronidase (GusA) reporters was achieved by single-copy chromosomal insertion via Tn5-mediated transposition. In addition, gene duplication by Mag-EGFP–EGFP fusions to MamC resulted in further increased magnetosome expression and fluorescence. Between 80 and 210 (for single MamC–Mag-EGFP) and 200 and 520 (for MamC–Mag-EGFP–EGFP) GFP copies were estimated to be expressed per individual magnetosome particle. PMID:24532068

  4. Expression of Human Carbonic Anhydrase in the Cyanobacterium Synechococcus PCC7942 Creates a High CO2-Requiring Phenotype 1

    PubMed Central

    Price, G. D.; Badger, M. R.

    1989-01-01

    Active human carbonic anhydrase II (HCAII) protein was expressed in the cyanobacterium Synechococcus PCC7942 by means of transformation with the bidirectional expression vector, pCA. This expression was driven by the bacterial Tac promoter and was regulated by the IacIQ repressor protein, which was expressed from the same plasmid. Expression levels reached values of around 0.3% of total cell protein and this protein appeared to be entirely soluble in nature and located within the cytosol of the cell. The expression of this protein has dramatic effects on the photosynthetic physiology of the cell. Induction of expression of carbonic anhydrase (CA) activity in both high dissolved inorganic carbon (Ci) and low Ci grown cells leads the creation of a high Ci requiring phenotype causing: (a) a dramatic increase in the K0.5 (Ci) for photosynthesis, (b) a loss of the ability to accumulate internal Ci, and (c) a decrease in the lag between the initial Ci accumulation following illumination and the efflux of CO2 from the cells. In addition, the effects of the expressed CA can largely be reversed by the carbonic anhydrase inhibitor ethoxyzolamide. As a result of the above findings, it is concluded that the CO2 concentrating mechanism in Synechococcus PCC7942 is largely dependent on (a) the absence of CA activity from the cytosol, and (b) the specific localization of CA activity in the carboxysome. A theoretical model of photosynthesis and Ci accumulation is developed in which the carboxysome plays a central role as both the site of CO2 generation from HCO3− and a resistance barrier to CO2 efflux from the cell. There is good qualitative agreement between this model and the measured physiological effects of expressed cytosolic CA in Synechococcus cells. Images Figure 7 PMID:16667062

  5. The perceptions of teachers and principals toward providing additional compensation to teachers in high-need subject areas

    NASA Astrophysics Data System (ADS)

    Longing, Jeffrey Lucian

    The purpose of this study was to determine possible differences in the perceptions of teachers teaching in high-need areas (i.e., math, science, special education, etc.) and teachers not teaching in high-need areas, (i.e., business education, physical education, etc.) as defined by the states of Arkansas and Louisiana, regarding higher compensation for high-need teachers. In addition, possible perception differences among principals and teachers were determined. The independent variables consisted of gender, position held, years of certified experience, and certification areas. The dependent variable was the perceptions of the participants on providing higher compensation for high-need teachers in order to attract and retain them. The data for all variables were collected using the Teacher Compensation Survey. The sample for this study was limited to teachers, grades 9 through 12, and principals of public high schools in south Arkansas and north Louisiana. Forty-four school districts in south Arkansas (Arkansas Department of Education, 2008a) and north Louisiana (Louisiana Department of Education, 2008a) met the criteria for this study. Twenty-two superintendents gave permission for their districts to participate in the research. A sample of 849 teachers and 38 principals were identified in these districts. Surveys were returned from 350 teachers, creating a 41% response rate. When the 31 principals that returned surveys were added to the total population, the response rate increased to 43% with 381 of the 887 surveyed responding. However, 42 of the teachers and two of the principals skipped some of the questions on the survey and were not included in the study. The researcher used a One-Way ANOVA and independent t-tests to determine the presence of statistical differences at the .05 level. The data showed that most math and science teachers agreed that high-need teachers should be compensated at a higher rate than teachers not teaching in high-need areas. The data

  6. Refinement of Eutectic Si in High Purity Al-5Si Alloys with Combined Ca and P Additions

    NASA Astrophysics Data System (ADS)

    Ludwig, Thomas Hartmut; Li, Jiehua; Schaffer, Paul Louis; Schumacher, Peter; Arnberg, Lars

    2015-01-01

    The effects of combined additions of Ca and P on the eutectic Si in a series of high purity Al-5 wt pct Si alloys have been investigated with the entrained droplet technique and complementary sets of conventional castings. Differential scanning calorimetry (DSC) and thermal analysis were used to investigate the eutectic droplet undercooling and the recalescence undercooling, respectively. Optical microscopy, SEM, EPMA, and TEM were employed to characterize the resultant microstructures. It was found that 250 ppm Ca addition to Al-5Si wt pct alloys with higher P contents leads to a significant increase of the eutectic droplet undercooling. For low or moderate cooling rates, the TEM results underline that Ca additions do not promote Si twinning. Thus, a higher twin density cannot be expected in Ca containing Al-Si alloys after, e.g., sand casting. Consequently, a refinement of the eutectic Si from coarse flake-like to fine plate-like structure, rather than a modification of the eutectic Si to a fibrous morphology, was achieved. This strongly indicates that the main purpose of Ca additions is to counteract the coarsening effect of the eutectic Si imposed by higher P concentrations. Significant multiple Si twinning was observed in melt-spun condition; however, this can be attributed to the higher cooling rate. After DSC heating (slow cooling), most of Si twins disappeared. Thus, the well-accepted impurity-induced twinning mechanism may be not valid in the case of Ca addition. The possible refinement mechanisms were discussed in terms of nucleation and growth of eutectic Si. We propose that the pre-eutectic Al2Si2Ca phase and preferential formation of Ca3P2 deactivate impurity particles, most likely AlP, poisoning the nucleation sites for eutectic Si.

  7. Reduction of salt in pork sausages by the addition of carrot fibre or potato starch and high pressure treatment.

    PubMed

    Grossi, Alberto; Søltoft-Jensen, Jakob; Knudsen, Jes Christian; Christensen, Mette; Orlien, Vibeke

    2012-12-01

    The combined effect of high pressure processing (HPP) (400, 600 and 800 MPa) and carrot fibre (CF) and potato starch (PS) on low salt (1.2%) pork sausages was investigated and compared with high (1.8%) salt sausages. Sausages had a marked increase in whitening with increasing content of fibre or starch, pressure level, and process temperature. The degree of redness was mainly affected by pressure level and heat treatment. An important finding regarding salt reduction was that the use of starch or fibre had more impact on textural properties than the level of salt since Young's modulus and strain at fracture were mainly affected by formulation and HPP. Water binding capacity of low salt sausages was improved to the same level as high salt sausages with HPP and addition of CF or PS particularly by the addition of PS which produced sausages with better sensory properties than CF. The sensory analysis showed that this approach is promising for producing low salt sausages. PMID:22682686

  8. Hypermethylation of the alternative AWT1 promoter in hematological malignancies is a highly specific marker for acute myeloid leukemias despite high expression levels

    PubMed Central

    2014-01-01

    Background Wilms tumor 1 (WT1) is over-expressed in numerous cancers with respect to normal cells, and has either a tumor suppressor or oncogenic role depending on cellular context. This gene is associated with numerous alternatively spliced transcripts, which initiate from two different unique first exons within the WT1 and the alternative (A)WT1 promoter intervals. Within the hematological system, WT1 expression is restricted to CD34+/CD38- cells and is undetectable after differentiation. Detectable expression of this gene is an excellent marker for minimal residual disease in acute myeloid leukemia (AML), but the underlying epigenetic alterations are unknown. Methods To determine the changes in the underlying epigenetic landscape responsible for this expression, we characterized expression, DNA methylation and histone modification profiles in 28 hematological cancer cell lines and confirmed the methylation signature in 356 cytogenetically well-characterized primary hematological malignancies. Results Despite high expression of WT1 and AWT1 transcripts in AML-derived cell lines, we observe robust hypermethylation of the AWT1 promoter and an epigenetic switch from a permissive to repressive chromatin structure between normal cells and AML cell lines. Subsequent methylation analysis in our primary leukemia and lymphoma cohort revealed that the epigenetic signature identified in cell lines is specific to myeloid-lineage malignancies, irrespective of underlying mutational status or translocation. In addition to being a highly specific marker for AML diagnosis (positive predictive value 100%; sensitivity 86.1%; negative predictive value 89.4%), we show that AWT1 hypermethylation also discriminates patients that relapse from those achieving complete remission after hematopoietic stem cell transplantation, with similar efficiency to WT1 expression profiling. Conclusions We describe a methylation signature of the AWT1 promoter CpG island that is a promising marker for

  9. Direct and efficient ethanol production from high-yielding rice using a Saccharomyces cerevisiae strain that express amylases.

    PubMed

    Yamada, Ryosuke; Yamakawa, Syun-Ichi; Tanaka, Tsutomu; Ogino, Chiaki; Fukuda, Hideki; Kondo, Akihiko

    2011-04-01

    Efficient ethanol producing yeast Saccharomyces cerevisiae cannot produce ethanol from raw starch directly. Thus the conventional ethanol production required expensive and complex process. In this study, we developed a direct and efficient ethanol production process from high-yielding rice harvested in Japan by using amylase expressing yeast without any pretreatment or addition of enzymes or nutrients. Ethanol productivity from high-yielding brown rice (1.1g/L/h) was about 5-fold higher than that obtained from purified raw corn starch (0.2g/L/h) when nutrients were added. Using an inoculum volume equivalent to 10% of the fermentation volume without any nutrient supplementation resulted in ethanol productivity and yield reaching 1.2g/L/h and 101%, respectively, in a 24-h period. High-yielding rice was demonstrated to be a suitable feedstock for bioethanol production. In addition, our polyploid amylase-expressing yeast was sufficiently robust to produce ethanol efficiently from real biomass. This is first report of direct ethanol production on real biomass using an amylase-expressing yeast strain without any pretreatment or commercial enzyme addition.

  10. Boronic Acid-Catalyzed, Highly Enantioselective Aza-Michael Additions of Hydroxamic Acid to Quinone Imine Ketals.

    PubMed

    Hashimoto, Takuya; Gálvez, Alberto Osuna; Maruoka, Keiji

    2015-12-30

    Boronic acid is one of the most versatile organic molecules in chemistry. Its uses include organic reactions, molecular recognition, assembly, and even medicine. While boronic acid catalysis, which utilizes an inherent catalytic property, has become an important research objective, it still lags far behind other boronic acid chemistries. Here, we report our discovery of a new boronic acid catalysis that enables the aza-Michael addition of hydroxamic acid to quinone imine ketals. By using 3-borono-BINOL as a chiral boronic acid catalyst, this reaction could be implemented in a highly enantioselective manner, paving the way to densely functionalized cyclohexanes.

  11. Highly E-Selective and Enantioselective Michael Addition to Electron-Deficient Internal Alkynes Under Chiral Iminophosphorane Catalysis.

    PubMed

    Uraguchi, Daisuke; Yamada, Kohei; Ooi, Takashi

    2015-08-17

    A highly E-selective and enantioselective conjugate addition of 2-benzyloxythiazol-5(4H)-ones to β-substituted alkynyl N-acyl pyrazoles is achieved under the catalysis of a P-spiro chiral iminophosphorane. Simultaneous control of the newly generated central chirality and olefin geometry is possible with a wide array of the alkynyl Michael acceptors possessing different aromatic and aliphatic β-substituents, as well as the various α-amino acid-derived thiazolone nucleophiles. This protocol provides access to structurally diverse, optically active α-amino acids bearing a geometrically defined trisubstituted olefinic component at the α-position.

  12. Effect of high energy β-radiation and addition of triallyl isocyanurate on the selected properties of polylactide

    NASA Astrophysics Data System (ADS)

    Malinowski, Rafał

    2016-06-01

    Comparison of some changes occurring in polylactide (PLA) due to high energy β-radiation and addition of triallyl isocyanurate (TAIC) was the main objective of the present study. It was found that irradiation of PLA by high energy β-radiation causes essential changes in its properties, that undergoes mainly degradation, to form a porous structure. The PLA degradation can be diminished by introduction into the polymer matrix of a low-molecular mass multifunctional compound like TAIC. Upon the electron radiation, effective crosslinking of PLA by TAIC occurs. Application of TAIC favorably influences hindering of the PLA degradation or, when the doses are very large, diminishes worsening of the PLA functional qualities. It was also found that the optimum crosslinking of PLA is obtained when the electron radiation doses of the range of 40-200 kGy are applied and the amount of TAIC equal 3-5 wt% is used.

  13. Surface-imprinted magnetic particles for highly selective sulfonamides recognition prepared by reversible addition fragmentation chain transfer polymerization.

    PubMed

    Xie, Xiaoyu; Liu, Xia; Pan, Xiaoyan; Chen, Liang; Wang, Sicen

    2016-01-01

    In this work, novel magnetic molecularly imprinted polymers (MMIPs) were prepared by reversible addition fragmentation chain transfer (RAFT) polymerization using sulfamerazine as the template. With the controlled/living property of RAFT polymerization, the resulting MMIPs showed high selectivity for sulfonamides recognition. The MMIPs were characterized by transmission electron microscopy, Fourier transform infrared, vibrating sample magnetometer, X-ray diffraction, X-ray photoelectron spectroscopy, and thermogravimetric analysis. The static and selectivity binding experiments demonstrated the desirable adsorption capacity and high selectivity of the MMIPs. The developed MMIPs were used as the solid-phase extraction sorbents to selectively extract four sulfonamides from aqueous solution. The recoveries of the spiked pond water ranged from 61.2 to 94.1% with RSD lower than 6.5%. This work demonstrated a versatile approach for the preparation of well-constructed MMIPs for application in the field of solid-phase extraction. PMID:26637219

  14. Layered Double Hydroxide Nanoplatelets with Excellent Tribological Properties under High Contact Pressure as Water-Based Lubricant Additives

    PubMed Central

    Wang, Hongdong; Liu, Yuhong; Chen, Zhe; Wu, Bibo; Xu, Sailong; Luo, Jianbin

    2016-01-01

    High efficient and sustainable utilization of water-based lubricant is essential for saving energy. In this paper, a kind of layered double hydroxide (LDH) nanoplatelets is synthesized and well dispersed in water due to the surface modification with oleylamine. The excellent tribological properties of the oleylamine-modified Ni-Al LDH (NiAl-LDH/OAm) nanoplatelets as water-based lubricant additives are evaluated by the tribological tests in an aqueous environment. The modified LDH nanoplatelets are found to not only reduce the friction but also enhance the wear resistance, compared with the water-based cutting fluid and lubricants containing other particle additives. By adding 0.5 wt% LDH nanoplatelets, under 1.5 GPa initial contact pressure, the friction coefficient, scar diameter, depth and width of the wear track dramatically decrease by 83.1%, 43.2%, 88.5% and 59.5%, respectively. It is considered that the sufficiently small size and the excellent dispersion of NiAl-LDH/OAm nanoplatelets in water are the key factors, so as to make them enter the contact area, form a lubricating film and prevent direct collision of asperity peaks. Our investigations demonstrate that the LDH nanoplatelet as a water-based lubricant additive has a great potential value in industrial application. PMID:26951794

  15. Layered Double Hydroxide Nanoplatelets with Excellent Tribological Properties under High Contact Pressure as Water-Based Lubricant Additives

    NASA Astrophysics Data System (ADS)

    Wang, Hongdong; Liu, Yuhong; Chen, Zhe; Wu, Bibo; Xu, Sailong; Luo, Jianbin

    2016-03-01

    High efficient and sustainable utilization of water-based lubricant is essential for saving energy. In this paper, a kind of layered double hydroxide (LDH) nanoplatelets is synthesized and well dispersed in water due to the surface modification with oleylamine. The excellent tribological properties of the oleylamine-modified Ni-Al LDH (NiAl-LDH/OAm) nanoplatelets as water-based lubricant additives are evaluated by the tribological tests in an aqueous environment. The modified LDH nanoplatelets are found to not only reduce the friction but also enhance the wear resistance, compared with the water-based cutting fluid and lubricants containing other particle additives. By adding 0.5 wt% LDH nanoplatelets, under 1.5 GPa initial contact pressure, the friction coefficient, scar diameter, depth and width of the wear track dramatically decrease by 83.1%, 43.2%, 88.5% and 59.5%, respectively. It is considered that the sufficiently small size and the excellent dispersion of NiAl-LDH/OAm nanoplatelets in water are the key factors, so as to make them enter the contact area, form a lubricating film and prevent direct collision of asperity peaks. Our investigations demonstrate that the LDH nanoplatelet as a water-based lubricant additive has a great potential value in industrial application.

  16. Layered Double Hydroxide Nanoplatelets with Excellent Tribological Properties under High Contact Pressure as Water-Based Lubricant Additives.

    PubMed

    Wang, Hongdong; Liu, Yuhong; Chen, Zhe; Wu, Bibo; Xu, Sailong; Luo, Jianbin

    2016-01-01

    High efficient and sustainable utilization of water-based lubricant is essential for saving energy. In this paper, a kind of layered double hydroxide (LDH) nanoplatelets is synthesized and well dispersed in water due to the surface modification with oleylamine. The excellent tribological properties of the oleylamine-modified Ni-Al LDH (NiAl-LDH/OAm) nanoplatelets as water-based lubricant additives are evaluated by the tribological tests in an aqueous environment. The modified LDH nanoplatelets are found to not only reduce the friction but also enhance the wear resistance, compared with the water-based cutting fluid and lubricants containing other particle additives. By adding 0.5 wt% LDH nanoplatelets, under 1.5 GPa initial contact pressure, the friction coefficient, scar diameter, depth and width of the wear track dramatically decrease by 83.1%, 43.2%, 88.5% and 59.5%, respectively. It is considered that the sufficiently small size and the excellent dispersion of NiAl-LDH/OAm nanoplatelets in water are the key factors, so as to make them enter the contact area, form a lubricating film and prevent direct collision of asperity peaks. Our investigations demonstrate that the LDH nanoplatelet as a water-based lubricant additive has a great potential value in industrial application. PMID:26951794

  17. Supplementing High-Density SNP Microarrays for Additional Coverage of Disease-Related Genes: Addiction as a Paradigm

    SciTech Connect

    SacconePhD, Scott F; Chesler, Elissa J; Bierut, Laura J; Kalivas, Peter J; Lerman, Caryn; Saccone, Nancy L; Uhl, George R; Li, Chuan-Yun; Philip, Vivek M; Edenberg, Howard; Sherry, Steven; Feolo, Michael; Moyzis, Robert K; Rutter, Joni L

    2009-01-01

    Commercial SNP microarrays now provide comprehensive and affordable coverage of the human genome. However, some diseases have biologically relevant genomic regions that may require additional coverage. Addiction, for example, is thought to be influenced by complex interactions among many relevant genes and pathways. We have assembled a list of 486 biologically relevant genes nominated by a panel of experts on addiction. We then added 424 genes that showed evidence of association with addiction phenotypes through mouse QTL mappings and gene co-expression analysis. We demonstrate that there are a substantial number of SNPs in these genes that are not well represented by commercial SNP platforms. We address this problem by introducing a publicly available SNP database for addiction. The database is annotated using numeric prioritization scores indicating the extent of biological relevance. The scores incorporate a number of factors such as SNP/gene functional properties (including synonymy and promoter regions), data from mouse systems genetics and measures of human/mouse evolutionary conservation. We then used HapMap genotyping data to determine if a SNP is tagged by a commercial microarray through linkage disequilibrium. This combination of biological prioritization scores and LD tagging annotation will enable addiction researchers to supplement commercial SNP microarrays to ensure comprehensive coverage of biologically relevant regions.

  18. High expression of small GTPase Rab3D promotes cancer progression and metastasis

    PubMed Central

    Yang, Jian; Liu, Wei; Lu, Xin'an; Fu, Yan; Li, Lin; Luo, Yongzhang

    2015-01-01

    Rab GTPases control exocytic and endocytic membrane trafficking such as exosomes release. As a secretory small GTPase, Rab3D is a vital regulator for protein secretion. However, the role of Rab3D in cancer was never systematically studied. The aim of this study is to examine its function and mechanism in cancer, especially metastasis. We detected protein levels of Rab3D in nine cancer cell lines and twelve types of clinical cancer specimens. Subsequently, we established in vitro migration and in vivo orthotopic metastatic mouse models to study the role of Rab3D in tumor metastasis. Here, we reported that the expression levels of Rab3D were dysregulated in cancer cells and highly correlated with tumor malignancies in the clinical samples. Increased expressions of Rab3D led to tumor invasion in vitro and lung metastasis in vivo, whereas Rab3D knockdown suppressed the tumor cell motility. Mechanistic studies revealed that Rab3D activated intracellular the AKT/GSK3β signaling to induce the EMT process. In addition, it also regulated the extracellular secretion of Hsp90α to promote tumor cell migration and invasion. These results prove that Rab3D is a key molecule to regulate tumor metastasis, suggesting that blocking the Rab3D function can be a potential therapeutic approach for cancer metastasis. PMID:25823663

  19. FXR Agonist INT-747 Upregulates DDAH Expression and Enhances Insulin Sensitivity in High-Salt Fed Dahl Rats

    PubMed Central

    Ghebremariam, Yohannes T.; Yamada, Keisuke; Lee, Jerry C.; Johnson, Christine L. C.; Atzler, Dorothee; Anderssohn, Maike; Agrawal, Rani; Higgins, John P.; Patterson, Andrew J.; Böger, Rainer H.; Cooke, John P

    2013-01-01

    Aims Genetic and pharmacological studies have shown that impairment of the nitric oxide (NO) synthase (NOS) pathway is associated with hypertension and insulin-resistance (IR). In addition, inhibition of NOS by the endogenous inhibitor, asymmetric dimethylarginine (ADMA), may also result in hypertension and IR. On the other hand, overexpression of dimethylarginine dimethylaminohydrolase (DDAH), an enzyme that metabolizes ADMA, in mice is associated with lower ADMA, increased NO and enhanced insulin sensitivity. Since DDAH carries a farnesoid X receptor (FXR)-responsive element, we aimed to upregulate its expression by an FXR-agonist, INT-747, and evaluate its effect on blood pressure and insulin sensitivity. Methods and Results In this study, we evaluated the in vivo effect of INT-747 on tissue DDAH expression and insulin sensitivity in the Dahl rat model of salt-sensitive hypertension and IR (Dahl-SS). Our data indicates that high salt (HS) diet significantly increased systemic blood pressure. In addition, HS diet downregulated tissue DDAH expression while INT-747 protected the loss in DDAH expression and enhanced insulin sensitivity compared to vehicle controls. Conclusion Our study may provide the basis for a new therapeutic approach for IR by modulating DDAH expression and/or activity using small molecules. PMID:23593273

  20. Venus Express bistatic radar: High-elevation anomalous reflectivity

    NASA Astrophysics Data System (ADS)

    Simpson, Richard A.; Tyler, G. Leonard; Häusler, Bernd; Mattei, Riccardo; Pätzold, Martin

    2009-06-01

    Magellan (MGN) bistatic radar observations in 1994 confirmed earlier Pioneer Venus reports of unusual Venus surface reflectivity and emissivity at elevations above 6054 km radius. They also revealed that the anomalous values of surface dielectric constant $\\varepsilon$ near Cleopatra Patera included a large imaginary component ($\\varepsilon$ ≈ -i 100) at 13 cm wavelength, consistent with a semiconducting surface material. The MGN observations were conducted using a linearly polarized wave, canted at 45° with respect to the plane of incidence and radiated by the MGN synthetic aperture radar antenna toward the specularly reflecting region of the mean planetary surface. In 2006 similar experiments were conducted using 13 cm circularly polarized transmissions from Venus Express (VEX). The VEX signal-to-noise ratio (SNR) was lower than that of MGN, but elevated ∣$\\varepsilon$∣ has been inferred broadly over Maxwell Montes. A quasi-specular echo was detected near Cleopatra but with insufficient SNR to address the question of conductivity. An early failure of the VEX 13 cm radio system precludes further measurements with VEX.

  1. Viscosity modification of high-oleic sunflower oil with polymeric additives for the design of new biolubricant formulations.

    PubMed

    Quinchia, L A; Delgado, M A; Valencia, C; Franco, J M; Gallegos, C

    2009-03-15

    Although most common lubricants contain mineral or synthetic oils as basestocks, new environmental regulations are demanding environmentally friendly lubricants. In this sense, vegetable oils represent promising alternatives to mineral-based lubricants because of their high biodegradability, good lubricity, and low volatility. However, their poor thermooxidative stability and the small range of viscosity represent a clear disadvantage to be used as suitable biolubricants. The main objective of this work was to develop new environmentally friendly lubricant formulations with improved kinematic viscosity values and viscosity thermal susceptibility. With this aim, a high-oleic sunflower oil (HOSO) was blended with polymeric additives, such as ethylene vinyl acetate (EVA) and styrene-butadiene-styrene (SBS) copolymers, at different concentrations (0.5-5% w/w). Dynamic viscosity and density measurements were performed in a rotational rheometer and capillary densimeter, respectively, in a temperature range between 25 and 120 degrees C. An Arrhenius-like equation fits the evolution of viscosity with temperature fairly well. Both EVA and SBS copolymers may be satisfactorily used as additives to increase the viscosity of HOSO, thus improving the low viscosity values of this oil. HOSO viscosity increases with polymer concentration. Specifically, EVA/HOSO blends exhibit higher viscosity values, which are needed for applications such as lubrication of bearings and four-stroke engines. On the other hand, viscositythermal susceptibility of HOSO samples increases with EVA or SBS concentration.

  2. Photocurable high internal phase emulsions (HIPEs) containing hydroxyapatite for additive manufacture of tissue engineering scaffolds with multi-scale porosity.

    PubMed

    Wang, Ai-Juan; Paterson, Thomas; Owen, Robert; Sherborne, Colin; Dugan, James; Li, Jun-Ming; Claeyssens, Frederik

    2016-10-01

    Porous composites containing hydroxyapatite (HA) were templated from high internal phase emulsions (HIPEs) and were further structured using direct-write UV stereolithography to produce composite scaffolds with multi-scale porosity. FTIR, TGA and SEM analyses confirmed that HA was retained after photocuring and subsequent treatments and was incorporated within the polymerised HIPE (polyHIPE). The addition of HA particles to the polyHIPE caused changes in the mechanical properties of the material. An increase in both the Young's modulus and maximum stress at yield was observed compared with the pure polyHIPE from 1.544±0.231 to 4.614±0.775 and 0.177±0.009 to 0.267±0.034MPa, respectively. Except at very high concentrations, adding HA did not adversely cause the phase separation of the HIPE or the porous microstructure of the resulting polyHIPE. In combination with a photoinitiator, the HIPE emulsion containing HA was investigated as a photocurable resin for stereolithography-based additive manufacturing. The material was readily processable into "woodpile" structures via direct-write UV stereolithography, producing scaffolds with multi-scale porosity which may be useful for medical applications such as tissue engineering. In conclusion, HA was successfully added into polyHIPEs, producing a similar porous structure to that of the pure polyHIPE whilst improving the mechanical performance. PMID:27287098

  3. Photocurable high internal phase emulsions (HIPEs) containing hydroxyapatite for additive manufacture of tissue engineering scaffolds with multi-scale porosity.

    PubMed

    Wang, Ai-Juan; Paterson, Thomas; Owen, Robert; Sherborne, Colin; Dugan, James; Li, Jun-Ming; Claeyssens, Frederik

    2016-10-01

    Porous composites containing hydroxyapatite (HA) were templated from high internal phase emulsions (HIPEs) and were further structured using direct-write UV stereolithography to produce composite scaffolds with multi-scale porosity. FTIR, TGA and SEM analyses confirmed that HA was retained after photocuring and subsequent treatments and was incorporated within the polymerised HIPE (polyHIPE). The addition of HA particles to the polyHIPE caused changes in the mechanical properties of the material. An increase in both the Young's modulus and maximum stress at yield was observed compared with the pure polyHIPE from 1.544±0.231 to 4.614±0.775 and 0.177±0.009 to 0.267±0.034MPa, respectively. Except at very high concentrations, adding HA did not adversely cause the phase separation of the HIPE or the porous microstructure of the resulting polyHIPE. In combination with a photoinitiator, the HIPE emulsion containing HA was investigated as a photocurable resin for stereolithography-based additive manufacturing. The material was readily processable into "woodpile" structures via direct-write UV stereolithography, producing scaffolds with multi-scale porosity which may be useful for medical applications such as tissue engineering. In conclusion, HA was successfully added into polyHIPEs, producing a similar porous structure to that of the pure polyHIPE whilst improving the mechanical performance.

  4. Additive effects of clofibric acid and pyruvate dehydrogenase kinase isoenzyme 4 (PDK4) deficiency on hepatic steatosis in mice fed a high-saturated fat diet

    PubMed Central

    Hwang, Byounghoon; Wu, Pengfei; Harris, Robert A.

    2012-01-01

    SUMMARY Although improving glucose metabolism by inhibition of pyruvate dehydrogenase kinase 4 (PDK4) might prove beneficial in the treatment of type 2 diabetes or diet-induced obesity, it might induce detrimental effects by inhibiting fatty acid oxidation. PPARα agonists are often used to treat dyslipidemia in patients, especially in type 2 diabetes. Combinational treatment with a PDK4 inhibitor and PPARα agonists may prove beneficial. However, PPARα agonists may be less effective in the presence of a PDK4 inhibitor because PPARα agonists induce PDK4 expression. In the present study, the effects of clofibric acid, a PPARα agonist, on blood and liver lipids were determined in wild type and PDK4 knockout mice fed a high fat diet. As expected, treatment of wild type mice with clofibric acid resulted in less body weight gain, smaller epididymal fat pads, greater insulin sensitivity, and lower levels of serum and liver triacylglycerol. Surprisingly, rather than decreasing the effectiveness of clofibric acid, PDK4 deficiency enhanced the beneficial effects of clofibric acid on hepatic steatosis, lowered blood glucose levels, and did not prevent the positive effects of clofibric acid on serum triacylglycerols and free fatty acids. The metabolic effects of clofibric acid are therefore independent of the induction of PDK4 expression. The additive beneficial effects on hepatic steatosis may be due to induction of increased capacity for fatty acid oxidation and partial uncoupling of oxidative phosphorylation by clofibric acid and a reduction in the capacity for fatty acid synthesis by PDK4 deficiency. PMID:22429297

  5. Gene expression profiles in testis of pigs with extreme high and low levels of androstenone

    PubMed Central

    Moe, Maren; Meuwissen, Theo; Lien, Sigbjørn; Bendixen, Christian; Wang, Xuefei; Conley, Lene Nagstrup; Berget, Ingunn; Tajet, Håvard; Grindflek, Eli

    2007-01-01

    Background: Boar taint is a major obstacle when using uncastrated male pigs for swine production. One of the main compounds causing this taint is androstenone, a pheromone produced in porcine testis. Here we use microarrays to study the expression of thousands of genes simultaneously in testis of high and low androstenone boars. The study allows identification of genes and pathways associated with elevated androstenone levels, which is essential for recognising potential molecular markers for breeding purposes. Results: Testicular tissue was collected from 60 boars, 30 with extreme high and 30 with extreme low levels of androstenone, from each of the two breeds Duroc and Norwegian Landrace. The samples were hybridised to porcine arrays containing 26,877 cDNA clones, detecting 563 and 160 genes that were differentially expressed (p < 0.01) in Duroc and Norwegian Landrace, respectively. Of these significantly up- and down-regulated clones, 72 were found to be common for the two breeds, suggesting the possibility of both general and breed specific mechanisms in regulation of, or response to androstenone levels in boars. Ten genes were chosen for verification of expression patterns by quantitative real competitive PCR and real-time PCR. As expected, our results point towards steroid hormone metabolism and biosynthesis as important biological processes for the androstenone levels, but other potential pathways were identified as well. Among these were oxidoreductase activity, ferric iron binding, iron ion binding and electron transport activities. Genes belonging to the cytochrome P450 and hydroxysteroid dehydrogenase families were highly up-regulated, in addition to several genes encoding different families of conjugation enzymes. Furthermore, a number of genes encoding transcription factors were found both up- and down-regulated. The high number of clones belonging to ferric iron and iron ion binding suggests an importance of these genes, and the association between

  6. High level expression of differentially localized BAG-1 isoforms in some oestrogen receptor-positive human breast cancers

    PubMed Central

    Brimmell, M; Burns, J S; Munson, P; McDonald, L; O’Hare, M J; Lakhani, S R; Packham, G

    1999-01-01

    Sensitivity to oestrogens and apoptosis are critical determinants of the development and progression of breast cancer and reflect closely linked pathways in breast epithelial cells. For example, induction of BCL-2 oncoprotein expression by oestrogen contributes to suppression of apoptosis and BCL-2 and oestrogen receptor (ER) are frequently co-expressed in tumours. BAG-1/HAP is a multifunctional protein which complexes with BCL-2 and steroid hormone receptors (including the ER), and can suppress apoptosis and influence steroid hormone-dependent transcription. Therefore, analysis of expression of BAG-1 in human breast cancer is of considerable interest. BAG-1 was readily detected by immunostaining in normal breast epithelial cells and most ER-positive tumours, but was undetectable or weakly expressed in ER-negative tumours. BAG-1 positive cells showed a predominantly cytoplasmic or cytoplasmic plus nuclear distribution of staining. A correlation between ER and BAG-1 was also evident in breast cancer derived cell lines, as all lines examined with functional ER expression also expressed high levels of BAG-1. In addition to the prototypical 36 kDa BAG-1 isoform, breast cancer cells expressed higher molecular weight isoforms and, in contrast to BCL-2, BAG-1 expression was independent of oestrogens. BAG-1 isoforms were differentially localized to the nucleus or cytoplasm and this was also independent of oestrogens. These results demonstrate a close association between BAG-1 and functional ER expression and suggest BAG-1 may be useful as a therapeutic target or prognostic marker in breast cancer. © 1999 Cancer Research Campaign PMID:10576663

  7. High-Affinity Glucose Transport in Aspergillus nidulans Is Mediated by the Products of Two Related but Differentially Expressed Genes

    PubMed Central

    Ventura, Luisa; González, Ramón; Ramón, Daniel; MacCabe, Andrew P.

    2014-01-01

    Independent systems of high and low affinity effect glucose uptake in the filamentous fungus Aspergillus nidulans. Low-affinity uptake is known to be mediated by the product of the mstE gene. In the current work two genes, mstA and mstC, have been identified that encode high-affinity glucose transporter proteins. These proteins' primary structures share over 90% similarity, indicating that the corresponding genes share a common origin. Whilst the function of the paralogous proteins is little changed, they differ notably in their patterns of expression. The mstC gene is expressed during the early phases of germination and is subject to CreA-mediated carbon catabolite repression whereas mstA is expressed as a culture tends toward carbon starvation. In addition, various pieces of genetic evidence strongly support allelism of mstC and the previously described locus sorA. Overall, our data define MstC/SorA as a high-affinity glucose transporter expressed in germinating conidia, and MstA as a high-affinity glucose transporter that operates in vegetative hyphae under conditions of carbon limitation. PMID:24751997

  8. High-affinity glucose transport in Aspergillus nidulans is mediated by the products of two related but differentially expressed genes.

    PubMed

    Forment, Josep V; Flipphi, Michel; Ventura, Luisa; González, Ramón; Ramón, Daniel; Maccabe, Andrew P

    2014-01-01

    Independent systems of high and low affinity effect glucose uptake in the filamentous fungus Aspergillus nidulans. Low-affinity uptake is known to be mediated by the product of the mstE gene. In the current work two genes, mstA and mstC, have been identified that encode high-affinity glucose transporter proteins. These proteins' primary structures share over 90% similarity, indicating that the corresponding genes share a common origin. Whilst the function of the paralogous proteins is little changed, they differ notably in their patterns of expression. The mstC gene is expressed during the early phases of germination and is subject to CreA-mediated carbon catabolite repression whereas mstA is expressed as a culture tends toward carbon starvation. In addition, various pieces of genetic evidence strongly support allelism of mstC and the previously described locus sorA. Overall, our data define MstC/SorA as a high-affinity glucose transporter expressed in germinating conidia, and MstA as a high-affinity glucose transporter that operates in vegetative hyphae under conditions of carbon limitation. PMID:24751997

  9. Characterization of LEF1 High Expression and Novel Mutations in Adult Acute Lymphoblastic Leukemia

    PubMed Central

    Liu, Juan; Li, Min; Song, Chunhua; Dovat, Sinisa; Li, Jianyong; Ge, Zheng

    2015-01-01

    Aberrant activation of the Wnt pathway plays a pathogenetic role in tumors and has been associated with adverse outcome in acute lymphoblastic leukemia (ALL). Lymphoid enhancer binding factor 1 (LEF1), a key mediator of Wnt signaling, has been linked to leukemic transformation, and LEF1 mutations have been identified in T-ALL. Here we found LEF1 is highly expressed in 25.0% adult ALL patients and LEF1 high expression was associated with high-risk leukemia factors (high WBC, Philadelphia chromosome positive, complex karyotype), shorter event-free survival (EFS), and high relapse rates in patients with B-ALL. LEF1 high expression is also associated with high mutation rate of Notch1 and JAK1 in T-ALL. We identified 2 novel LEF1 mutations (K86E and P106L) in 4 of 131 patients with ALL, and those patients with high-risk ALL (high WBC, complex karyotype). These results suggest a role for LEF1 mutations in leukemogenesis. We further explored the effect of the mutations on cell proliferation and found both mutations significantly promoted the proliferation of ALL cells. We also observed the effect of LEF1 and its mutations on the transcription of its targets, c-MYC and Cyclin D1. We found LEF1 increased the promoter activity of its targets c-MYC and Cyclin D1, and LEF1 K86E and P106L mutants further significantly enhanced this effect. We also observed that the c-MYC and Cyclin D1 mRNA levels were significantly increased in patients with LEF1 high expression compared with those with low expression. Taken together, our findings indicate high LEF1 expression and mutation are associated with high-risk leukemia and our results also revealed that LEF1 high expression and/or gain-of-function mutations are involved in leukemogenesis of ALL. PMID:25942645

  10. Freedom of Expression Laws and the College Press: Lessons Learned from the High Schools.

    ERIC Educational Resources Information Center

    Paxton, Mark

    This paper examines two recent attempts to enact state freedom of expression laws for public college and university students and discusses the prospects for such laws in the context of state scholastic freedom of expression laws covering high school journalists in six states. It examines the case of Kincaid v. Gibson, which decided that…

  11. Adaptation of the highly productive T7 expression system to Streptomyces lividans.

    PubMed

    Lussier, François-Xavier; Denis, François; Shareck, François

    2010-02-01

    Streptomyces lividans is a Gram-positive bacterium known for its remarkable secretion efficiency and low extracellular protease activity. In the present work, we adapted the highly productive T7 expression system to S. lividans. A codon-optimized T7 RNA polymerase gene was chromosomally integrated, and a bifunctional T7 expression vector was constructed.

  12. Apolipoprotein E expression and behavioral toxicity of high charge, high energy (HZE) particle radiation

    NASA Technical Reports Server (NTRS)

    Higuchi, Yoshinori; Nelson, Gregory A.; Vazquez, Marcelo; Laskowitz, Daniel T.; Slater, James M.; Pearlstein, Robert D.

    2002-01-01

    Apolipoprotein E (apoE) is a lipid binding protein that plays an important role in tissue repair following brain injury. In the present studies, we have investigated whether apoE affects the behavioral toxicity of high charge, high energy (HZE) particle radiation. METHODS: Sixteen male apoE knockout (KO) mice and sixteen genetically matched wild-type (WT) C57BL mice were used in this experiment. Half of the KO and half of the WT animals were irradiated with 600 MeV/amu iron particles (2 Gy whole body). The effect of irradiation on motor coordination and stamina (Rotarod test), exploratory behavior (open field test), and spatial working and reference memory (Morris water maze) was assessed. ROTAROD TEST: Performance was adversely affected by radiation exposure in both KO and WT groups at 30 d after irradiation. By 60 d after radiation, the radiation effect was lost in WT, but still apparent in irradiated KO mice. OPEN FIELD TEST: Radiation reduced open field exploratory activity 14, 28, 56, 84, and 168 d after irradiation of KO mice, but had no effect on WT mice. MORRIS WATER MAZE: Radiation adversely affected spatial working memory in the KO mice, but had no discernible effect in the WT mice as assessed 180 d after irradiation. In contrast, irradiated WT mice showed marked impairment of spatial reference memory in comparison to non-irradiated mice, while no effect of radiation was observed in KO mice. CONCLUSIONS: These studies show that apoE expression influences the behavioral toxicity of HZE particle radiation and suggest that apoE plays a role in the repair/recovery from radiation injury of the CNS. ApoE deficiency may exacerbate the previously reported effects of HZE particle radiation in accelerating the brain aging process.

  13. Expression Profiling during Arabidopsis/Downy Mildew Interaction Reveals a Highly-Expressed Effector That Attenuates Responses to Salicylic Acid

    PubMed Central

    Asai, Shuta; Caillaud, Marie-Cécile; Furzer, Oliver J.; Ishaque, Naveed; Wirthmueller, Lennart; Fabro, Georgina; Shirasu, Ken; Jones, Jonathan D. G.

    2014-01-01

    Plants have evolved strong innate immunity mechanisms, but successful pathogens evade or suppress plant immunity via effectors delivered into the plant cell. Hyaloperonospora arabidopsidis (Hpa) causes downy mildew on Arabidopsis thaliana, and a genome sequence is available for isolate Emoy2. Here, we exploit the availability of genome sequences for Hpa and Arabidopsis to measure gene-expression changes in both Hpa and Arabidopsis simultaneously during infection. Using a high-throughput cDNA tag sequencing method, we reveal expression patterns of Hpa predicted effectors and Arabidopsis genes in compatible and incompatible interactions, and promoter elements associated with Hpa genes expressed during infection. By resequencing Hpa isolate Waco9, we found it evades Arabidopsis resistance gene RPP1 through deletion of the cognate recognized effector ATR1. Arabidopsis salicylic acid (SA)-responsive genes including PR1 were activated not only at early time points in the incompatible interaction but also at late time points in the compatible interaction. By histochemical analysis, we found that Hpa suppresses SA-inducible PR1 expression, specifically in the haustoriated cells into which host-translocated effectors are delivered, but not in non-haustoriated adjacent cells. Finally, we found a highly-expressed Hpa effector candidate that suppresses responsiveness to SA. As this approach can be easily applied to host-pathogen interactions for which both host and pathogen genome sequences are available, this work opens the door towards transcriptome studies in infection biology that should help unravel pathogen infection strategies and the mechanisms by which host defense responses are overcome. PMID:25329884

  14. A novel highly thermostable xylanase stimulated by Ca2+ from Thermotoga thermarum: cloning, expression and characterization

    PubMed Central

    2013-01-01

    Background Xylanase is an important component of hemicellulase enzyme system. Since it plays an important role in the hydrolysis of hemicellulose into xylooligosaccharides (XOs), high thermostable xylanase has been the focus of much recent attention as powerful enzyme as well as in the field of biomass utilization. Results A xylanase gene (xyn10A) with 3,474 bp was cloned from the extremely thermophilic bacterium Thermotoga thermarum that encodes a protein containing 1,158 amino acid residues. Based on amino acid sequence homology, hydrophobic cluster and three dimensional structure analyses, it was attested that the xylanase belongs to the glycoside hydrolase (GH) families 10 with five carbohydrate binding domains. When the xylanase gene was cloned and expressed in Escherichia coli BL21 (DE3), the specific enzyme activity of xylanase produced by the recombinant strain was up to 145.8 U mg-1. The xylanase was optimally active at 95°C, pH 7.0. In addition, it exhibited high thermostability over broad range of pH 4.0-8.5 and temperature 55-90°C upon the addition of 5 mM Ca2+. Confirmed by Ion Chromatography System (ICS) analysis, the end products of the hydrolysis of beechwood xylan were xylose, xylobiose, xylotriose, xylotetraose, xylopentaose and xylohexaose. Conclusions The xylanase from T. thermarum is one of the hyperthermophilic xylanases that exhibits high thermostability, and thus, is a suitable candidate for generating XOs from cellulosic materials such as agricultural and forestry residues for the uses as prebiotics and precursors for further preparation of furfural and other chemicals. PMID:23418789

  15. High quality Y3Al5O12 doped transparent ceramics for laser applications, role of sintering additives

    NASA Astrophysics Data System (ADS)

    Kaminskii, A. A.; Balashov, V. V.; Cheshev, E. A.; Kopylov, Yu L.; Koromyslov, A. L.; Krokhin, O. N.; Kravchenko, V. B.; Lopukhin, K. V.; Shemet, V. V.; Tupitsyn, I. M.

    2016-08-01

    SiO2, ZrO2, B2O3 and MgO oxides and their combinations were used as sintering aids for preparation of yttrium aluminum garnet (YAG) ceramics doped by Nd2O3, Er2O3, Ho2O3, Tm2O3 and Yb2O3. The influence of these additives on optimal sintering temperature, grain growth, volume of residual pores and optical quality of the ceramics were investigated. The best combination of the sintering additives was found and high quality samples of YAG:Nd (1 at.%) ceramics were obtained. The original method of laser optical quality characterization of ceramics was developed and tested. The main laser parameters of YAG:Nd (1 at.%) ceramics samples are measured and compared with the best well known laser ceramics. The samples of YAG:RE (RE- Er2O3, Ho2O3, Tm2O3 and Yb2O3) ceramics are obtained, and their optical transmittance spectra are measured. Composite structures of YAG:Yb (5 at.%) - YAG were obtained by the simplest method of successive joint compaction of different composition layers.

  16. Graphene oxide as a high-performance fluid-loss-control additive in water-based drilling fluids.

    PubMed

    Kosynkin, Dmitry V; Ceriotti, Gabriel; Wilson, Kurt C; Lomeda, Jay R; Scorsone, Jason T; Patel, Arvind D; Friedheim, James E; Tour, James M

    2012-01-01

    Graphene oxide (GO) performs well as a filtration additive in water-based drilling fluids at concentrations as low as 0.2 % (w/w) by carbon content. Standard American Petroleum Institute (API) filtration tests were conducted on pH-adjusted, aqueous dispersions of GO and xanthan gum. It was found that a combination of large-flake GO and powdered GO in a 3:1 ratio performed best in the API tests, allowing an average fluid loss of 6.1 mL over 30 min and leaving a filter cake ~20 μm thick. In comparison, a standard suspension (~12 g/L) of clays and polymers used in the oil industry gave an average fluid loss of 7.2 mL and a filter cake ~280 μm thick. Scanning electron microscopy imaging revealed the extreme pliability of well-exfoliated GO, as the pressure due to filtration crumpled single GO sheets, forcing them to slide through pores with diameters much smaller than the flake's flattened size. GO solutions also exhibited greater shear thinning and higher temperature stability compared to clay-based fluid-loss additives, demonstrating potential for high-temperature well applications.

  17. Graphene oxide as a high-performance fluid-loss-control additive in water-based drilling fluids.

    PubMed

    Kosynkin, Dmitry V; Ceriotti, Gabriel; Wilson, Kurt C; Lomeda, Jay R; Scorsone, Jason T; Patel, Arvind D; Friedheim, James E; Tour, James M

    2012-01-01

    Graphene oxide (GO) performs well as a filtration additive in water-based drilling fluids at concentrations as low as 0.2 % (w/w) by carbon content. Standard American Petroleum Institute (API) filtration tests were conducted on pH-adjusted, aqueous dispersions of GO and xanthan gum. It was found that a combination of large-flake GO and powdered GO in a 3:1 ratio performed best in the API tests, allowing an average fluid loss of 6.1 mL over 30 min and leaving a filter cake ~20 μm thick. In comparison, a standard suspension (~12 g/L) of clays and polymers used in the oil industry gave an average fluid loss of 7.2 mL and a filter cake ~280 μm thick. Scanning electron microscopy imaging revealed the extreme pliability of well-exfoliated GO, as the pressure due to filtration crumpled single GO sheets, forcing them to slide through pores with diameters much smaller than the flake's flattened size. GO solutions also exhibited greater shear thinning and higher temperature stability compared to clay-based fluid-loss additives, demonstrating potential for high-temperature well applications. PMID:22136134

  18. [Determination of arsanilic acid and sulfanilic acid as adulterant in feed additives by reversed-phase high performance liquid chromatography].

    PubMed

    Xu, Jinping; He, Heng; Xu, Mengyi; Qu, Yanhua

    2010-02-01

    A reversed-phase high performance liquid chromatographic (RP-HPLC) method was established for the determination of arsanilic acid and sulfanilic acid as adulterant in the feed additives. The separation was carried out on a Waters Bondapak C18 column, and methanol-water (pH 2.9 adjusted by 0.01 mol/L phosphoric acid) (1 : 4, v/v) was used as the mobile phase with a flow rate of 1.0 mL/min. A diode array detector was used at 244 nm as the detection wavelength. Arsanilic acid and sulfanilic acid were separated within 3 min. The linear ranges all were 5 - 200 mg/L and the detection limits (S/N = 3) were 0.20 and 0.15 mg/L for arsanilic acid and sulfanilic acid, respectively. This method is simple and rapid, and suitable for the simultaneous determination of arsanilic acid and sulfanilic acid in feed additives.

  19. Boric acid as a mobile phase additive for high performance liquid chromatography separation of ribose, arabinose and ribulose.

    PubMed

    De Muynck, Cassandra; Beauprez, Joeri; Soetaert, Wim; Vandamme, Erick J

    2006-01-01

    A new high performance liquid chromatographic (HPLC) method is described for the analysis of ribose, arabinose and ribulose mixtures obtained from (bio)chemical isomerization processes. These processes gain importance since the molecules can be used for the synthesis of antiviral therapeutics. The HPLC method uses boric acid as a mobile phase additive to enhance the separation on an Aminex HPX-87K column. By complexing with boric acid, the carbohydrates become negatively charged, thus elute faster from the column by means of ion exlusion and are separated because the complexation capacity with boric acid differs from one carbohydrate to another. Excellent separation between ribose, ribulose and arabinose was achieved with concentrations between 0.1 and 10 gL(-1) of discrete sugar.

  20. Meta-analysis of high-latitude nitrogen-addition and warming studies implies ecological mechanisms overlooked by land models

    DOE PAGES

    Bouskill, N. J.; Riley, W. J.; Tang, J. Y.

    2014-12-11

    Accurate representation of ecosystem processes in land models is crucial for reducing predictive uncertainty in energy and greenhouse gas feedbacks with the climate. Here we describe an observational and modeling meta-analysis approach to benchmark land models, and apply the method to the land model CLM4.5 with two versions of belowground biogeochemistry. We focused our analysis on the aboveground and belowground responses to warming and nitrogen addition in high-latitude ecosystems, and identified absent or poorly parameterized mechanisms in CLM4.5. While the two model versions predicted similar soil carbon stock trajectories following both warming and nitrogen addition, other predicted variables (e.g., belowgroundmore » respiration) differed from observations in both magnitude and direction, indicating that CLM4.5 has inadequate underlying mechanisms for representing high-latitude ecosystems. On the basis of observational synthesis, we attribute the model–observation differences to missing representations of microbial dynamics, aboveground and belowground coupling, and nutrient cycling, and we use the observational meta-analysis to discuss potential approaches to improving the current models. However, we also urge caution concerning the selection of data sets and experiments for meta-analysis. For example, the concentrations of nitrogen applied in the synthesized field experiments (average = 72 kg ha-1 yr-1) are many times higher than projected soil nitrogen concentrations (from nitrogen deposition and release during mineralization), which precludes a rigorous evaluation of the model responses to likely nitrogen perturbations. Overall, we demonstrate that elucidating ecological mechanisms via meta-analysis can identify deficiencies in ecosystem models and empirical experiments.« less

  1. Amylase addition increases starch ruminal digestion in first-lactation cows fed high and low starch diets.

    PubMed

    Nozière, P; Steinberg, W; Silberberg, M; Morgavi, D P

    2014-01-01

    The objective of this study was to evaluate the effect of an exogenous amylase preparation on digestion of low- and high-starch diets in dairy cattle. Rumen and total-tract nutrient digestibility were measured in a 4×4 Latin square design with 28-d periods using 4 first-lactation cows cannulated at the rumen and duodenum. Corn silage-based diets had 20 or 30% starch, attained by changing the composition of concentrate, with or without addition of an exogenous amylase preparation. Effects of the enzyme additive were observed on ruminal digestibility but not at the total-tract level. Ruminal digestibility of starch increased from 75% in control to 81% with amylase supplementation. This difference in ruminal starch digestion was compensated postruminally, so that the total-tract digestibility of starch was almost complete and did not differ between treatments. The amylase supplement also increased the true ruminal digestibility of organic matter but did not affect microbial N flow to the duodenum. Amylase supplement reduced the proportion of acetate and butyrate and increased that of propionate, particularly in the high-starch diet, where it tended to increase the concentration of total volatile fatty acids in the rumen. Other effects were a higher amylase activity in the solid-associated microbial community and a tendency for lower numbers of protozoa. In contrast, we observed no changes in intake, production, dry matter and fiber (neutral detergent fiber and acid detergent fiber) digestibility, or ruminal digestion, and no or small changes on selected fibrolytic and amylolytic bacteria and on the microbial community in general. We conclude that the exogenous amylase improved starch digestion in the rumen in first-lactation cows with moderate intake and production levels.

  2. A novel highly differentially expressed gene in wheat endosperm associated with bread quality

    PubMed Central

    Furtado, A.; Bundock, P. C.; Banks, P. M.; Fox, G.; Yin, X.; Henry, R. J.

    2015-01-01

    Analysis of gene expression in developing wheat seeds was used to identify a gene, wheat bread making (wbm), with highly differential expression (~1000 fold) in the starchy endosperm of genotypes varying in bread making quality. Several alleles differing in the 5’-upstream region (promoter) of this gene were identified, with one present only in genotypes with high levels of wbm expression. RNA-Seq analysis revealed low or no wbm expression in most genotypes but high expression (0.2-0.4% of total gene expression) in genotypes that had good bread loaf volume. The wbm gene is predicted to encode a mature protein of 48 amino acids (including four cysteine residues) not previously identified in association with wheat quality, possibly because of its small size and low frequency in the wheat gene pool. Genotypes with high wbm expression all had good bread making quality but not always good physical dough qualities. The predicted protein was sulphur rich suggesting the possibility of a contribution to bread loaf volume by supporting the crossing linking of proteins in gluten. Improved understanding of the molecular basis of differences in bread making quality may allow more rapid development of high performing genotypes with acceptable end-use properties and facilitate increased wheat production. PMID:26011437

  3. Sigma-1 receptor expression in the dorsal root ganglion: Reexamination using a highly specific antibody.

    PubMed

    Mavlyutov, Timur A; Duellman, Tyler; Kim, Hung Tae; Epstein, Miles L; Leese, Charlotte; Davletov, Bazbek A; Yang, Jay

    2016-09-01

    Sigma-1 receptor (S1R) is a unique pluripotent modulator of living systems and has been reported to be associated with a number of neurological diseases including pathological pain. Intrathecal administration of S1R antagonists attenuates the pain behavior of rodents in both inflammatory and neuropathic pain models. However, the S1R localization in the spinal cord shows a selective ventral horn motor neuron distribution, suggesting the high likelihood of S1R in the dorsal root ganglion (DRG) mediating the pain relief by intrathecally administered drugs. Since primary afferents are the major component in the pain pathway, we examined the mouse and rat DRGs for the presence of the S1R. At both mRNA and protein levels, quantitative RT-PCR (qRT-PCR) and Western confirmed that the DRG contains greater S1R expression in comparison to spinal cord, cortex, or lung but less than liver. Using a custom-made highly specific antibody, we demonstrated the presence of a strong S1R immuno-fluorescence in all rat and mouse DRG neurons co-localizing with the Neuron-Specific Enolase (NSE) marker, but not in neural processes or GFAP-positive glial satellite cells. In addition, S1R was absent in afferent terminals in the skin and in the dorsal horn of the spinal cord. Using immuno-electron microscopy, we showed that S1R is detected in the nuclear envelope and endoplasmic reticulum (ER) of DRG cells. In contrast to other cells, S1R is also located directly at the plasma membrane of the DRG neurons. The presence of S1R in the nuclear envelope of all DRG neurons suggests an exciting potential role of S1R as a regulator of neuronal nuclear activities and/or gene expression, which may provide insight toward new molecular targets for modulating nociception at the level of primary afferent neurons. PMID:27339730

  4. Sigma-1 receptor expression in the dorsal root ganglion: Reexamination using a highly specific antibody.

    PubMed

    Mavlyutov, Timur A; Duellman, Tyler; Kim, Hung Tae; Epstein, Miles L; Leese, Charlotte; Davletov, Bazbek A; Yang, Jay

    2016-09-01

    Sigma-1 receptor (S1R) is a unique pluripotent modulator of living systems and has been reported to be associated with a number of neurological diseases including pathological pain. Intrathecal administration of S1R antagonists attenuates the pain behavior of rodents in both inflammatory and neuropathic pain models. However, the S1R localization in the spinal cord shows a selective ventral horn motor neuron distribution, suggesting the high likelihood of S1R in the dorsal root ganglion (DRG) mediating the pain relief by intrathecally administered drugs. Since primary afferents are the major component in the pain pathway, we examined the mouse and rat DRGs for the presence of the S1R. At both mRNA and protein levels, quantitative RT-PCR (qRT-PCR) and Western confirmed that the DRG contains greater S1R expression in comparison to spinal cord, cortex, or lung but less than liver. Using a custom-made highly specific antibody, we demonstrated the presence of a strong S1R immuno-fluorescence in all rat and mouse DRG neurons co-localizing with the Neuron-Specific Enolase (NSE) marker, but not in neural processes or GFAP-positive glial satellite cells. In addition, S1R was absent in afferent terminals in the skin and in the dorsal horn of the spinal cord. Using immuno-electron microscopy, we showed that S1R is detected in the nuclear envelope and endoplasmic reticulum (ER) of DRG cells. In contrast to other cells, S1R is also located directly at the plasma membrane of the DRG neurons. The presence of S1R in the nuclear envelope of all DRG neurons suggests an exciting potential role of S1R as a regulator of neuronal nuclear activities and/or gene expression, which may provide insight toward new molecular targets for modulating nociception at the level of primary afferent neurons.

  5. Engineering the Expression and Characterization of Two Novel Laccase Isoenzymes from Coprinus comatus in Pichia pastoris by Fusing an Additional Ten Amino Acids Tag at N-Terminus

    PubMed Central

    Gu, Chunjuan; Zheng, Fei; Long, Liangkun; Wang, Jing; Ding, Shaojun

    2014-01-01

    The detail understanding of physiological/biochemical characteristics of individual laccase isoenzymes in fungi is necessary for fundamental and application purposes, but our knowledge is still limited for most of fungi due to difficult to express laccases heterologously. In this study, two novel laccase genes, named lac3 and lac4, encoding proteins of 547 and 532-amino acids preceded by 28 and 16-residue signal peptides, respectively, were cloned from the edible basidiomycete Coprinus comatus. They showed 70% identity but much lower homology with other fungal laccases at protein level (less than 58%). Two novel laccase isoenzymes were successfully expressed in Pichia pastoris by fusing an additional 10 amino acids (Thr-Pro-Phe-Pro-Pro-Phe-Asn-Thr-Asn-Ser) tag at N-terminus, and the volumetric activities could be dramatically enhanced from undetectable level to 689 and 1465 IU/l for Lac3 and Lac4, respectively. Both laccases possessed the lowest Km and highest kcat/Km value towards syringaldazine, followed by ABTS, guaiacol and 2,6-dimethylphenol similar as the low redox potential laccases from other microorganisms. Lac3 and Lac4 showed resistant to SDS, and retained 31.86% and 43.08% activity in the presence of 100 mM SDS, respectively. Lac3 exhibited higher decolorization efficiency than Lac4 for eleven out of thirteen different dyes, which may attribute to the relatively higher catalytic efficiency of Lac3 than Lac4 (in terms of kcat/Km) towards syringaldazine and ABTS. The mild synergistic decolorization by two laccases was observed for triphenylmethane dyes but not for anthraquinone and azo dyes. PMID:24710109

  6. Enhancement of β-carotene production by over-expression of HMG-CoA reductase coupled with addition of ergosterol biosynthesis inhibitors in recombinant Saccharomyces cerevisiae.

    PubMed

    Yan, Guo-liang; Wen, Ke-rui; Duan, Chang-qing

    2012-02-01

    In this study, the synergistic effect of overexpressing the 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase gene and adding ergosterol synthesis inhibitor, ketoconazole, on β-carotene production in the recombinant Saccharomyces cerevisiae was investigated. The results showed that the over-expression of HMG-CoA reductase gene and adding 100 mg/l ketoconazole alone can result in 135.1 and 15.6% increment of β-carotene concentration compared with that of the control (2.05 mg/g dry weight of cells), respectively. However, the combination of overexpressing HMG-CoA reductase gene and adding ketoconazole can achieve a 206.8% increment of pigment content (6.29 mg/g dry weight of cells) compared with that of the control. Due to the fact that over-expression of the HMG-CoA reductase gene can simultaneously improve the flux of the sterol and carotenoid biosynthetic pathway, it can be concluded that under the circumstances of blocking sterol biosynthesis, increasing the activity of HMG-CoA reductase can result in more precursors FPP fluxing into carotenoid branch and obtain a high increment of β-carotene production. The results of this study collectively suggest that the combination of overexpressing HMG-CoA reductase gene and supplying ergosterol synthesis inhibitor is an effective strategy to improve the production of desirable isoprenoid compounds such as carotenoids. PMID:22086347

  7. Identification of cis-elements conferring high levels of gene expression in non-green plastids.

    PubMed

    Zhang, Jiang; Ruf, Stephanie; Hasse, Claudia; Childs, Liam; Scharff, Lars B; Bock, Ralph

    2012-10-01

    Although our knowledge about the mechanisms of gene expression in chloroplasts has increased substantially over the past decades, next to nothing is known about the signals and factors that govern expression of the plastid genome in non-green tissues. Here we report the development of a quantitative method suitable for determining the activity of cis-acting elements for gene expression in non-green plastids. The in vivo assay is based on stable transformation of the plastid genome and the discovery that root length upon seedling growth in the presence of the plastid translational inhibitor kanamycin is directly proportional to the expression strength of the resistance gene nptII in transgenic tobacco plastids. By testing various combinations of promoters and translation initiation signals, we have used this experimental system to identify cis-elements that are highly active in non-green plastids. Surprisingly, heterologous expression elements from maize plastids were significantly more efficient in conferring high expression levels in root plastids than homologous expression elements from tobacco. Our work has established a quantitative method for characterization of gene expression in non-green plastid types, and has led to identification of cis-elements for efficient plastid transgene expression in non-green tissues, which are valuable tools for future transplastomic studies in basic and applied research.

  8. PTK7 regulates Id1 expression in CD44-high glioma cells

    PubMed Central

    Liu, Qing; Zhang, Chi; Yuan, Jian; Fu, Jun; Wu, Minghua; Su, Jun; Wang, Xiangyu; Yuan, Xianrui; Jiang, Weixi

    2015-01-01

    Background CD44 is a molecular marker associated with molecular subtype and treatment resistance in glioma. More effective therapies will result from approaches aimed at targeting the CD44-high gliomas. Methods Protein tyrosine kinase 7 (PTK7) mRNA expression was analyzed based on The Cancer Genome Atlas glioblastoma dataset. PTK7 expression was depleted through lentivirus-mediated short hairpin RNA knockdown. Terminal deoxynucleotidyl transferase dUTP nick-end labeling was used to evaluate cell apoptosis following PTK7 knockdown. Gene expression analysis was performed on Affymetrix microarray. A nude mice orthotopic tumor model was used to evaluate the in vivo effect of PTK7 depletion. Results PTK7 is highly expressed in CD44-high glioblastoma and predicts unfavorable prognosis. PTK7 knockdown attenuated cell proliferation, impaired tumorigenic potential, and induced apoptosis in CD44-high glioma cell lines. Gene expression analysis identified inhibitor of DNA Binding 1 (Id1) gene as a potential downstream effector for PTK7. Overexpression of Id1 mostly restored the cell proliferation and colony formation attenuated by PTK7 depletion. PTK7 enhanced anchorage-independent growth in normal human astrocytes, which was attenuated by Id1 knockdown. Furthermore, PTK7 regulated Id1 expression through modulating TGF-β/Smad signaling, while pharmacological inhibition on TGF-β/Smad signaling or PTK7/Id1 depletion attenuated TGF-β–stimulated cell proliferation. PTK7 depletion consistently reduced Id1 expression, suppressed tumor growth, and induced apoptosis in a murine orthotopic tumor model, which could be translated into prolonged survival in tumor-bearing mice. Conclusions PTK7 regulates Id1 expression in CD44-high glioma cell lines. Targeting PTK7 could be an effective strategy for treating glioma with high CD44 expression. PMID:25204555

  9. High Immunoglobulin A Levels Mediate the Association Between High Anger Expression and Low Somatic Symptoms in Intimate Partner Violence Perpetrators.

    PubMed

    Romero-Martínez, A; Lila, M; Vitoria-Estruch, S; Moya-Albiol, L

    2016-02-01

    It has been hypothesized that anger expression may be associated with increased salivary immunoglobulin A (sIgA) levels, which is associated with decreased somatic symptoms, and therefore anger expression may be associated with reduced somatic symptoms in intimate partner violence (IPV) perpetrators. This study tested the potential mediating effect of sIgA levels on the relationship between anger expression and respiratory and gastrointestinal symptoms in IPV perpetrators and non-violent controls. The sample consisted of IPV perpetrators (n = 19) and controls (n = 21). Saliva samples were collected for assessing sIgA levels. The State-Trait Anger Expression Inventory-2 was used to assess anger expression and the Revised version of the Somatic Symptoms Scale developed by Sandín and Chorot to measure somatic symptoms. High anger expression was associated with low levels of respiratory and gastrointestinal symptoms in IPV perpetrators mediated through high sIgA levels but the same was not true for non-violent controls. This finding supports the hypothesis that for IPV perpetrators, anger expression may be physiologically and psychologically rewarding. Future research examining other immunological parameters is needed to further test this hypothesis. Such effort may illuminate why some IPV perpetrators continue to use violence against their partners.

  10. Suppression of Ripening-Associated Gene Expression in Tomato Fruits Subjected to a High CO2 Concentration.

    PubMed Central

    Rothan, C.; Duret, S.; Chevalier, C.; Raymond, P.

    1997-01-01

    High concentrations of CO2 block or delay the ripening of fruits. In this study we investigated the effects of high CO2 on ripening and on the expression of stress- and ripening-inducible genes in cherry tomato (Lycopersicon esculentum Mill.) fruit. Mature-green tomato fruits were submitted to a high CO2 concentration (20%) for 3 d and then transferred to air. These conditions effectively inhibited ripening-associated color changes and ethylene production, and reduced the protein content. No clear-cut effect was observed on the expression of two proteolysis-related genes, encoding polyubiquitin and ubiquitin-conjugating enzyme E2, respectively. Exposure of fruit to high CO2 also resulted in the strong induction of two genes encoding stress-related proteins: a ripening-regulated heat-shock protein and glutamate decarboxylase. Induction of these two genes indicated that high CO2 had a stress effect, most likely through cytosolic acidification. In addition, high CO2 blocked the accumulation of mRNAs for genes involved in the main ripening-related changes: ethylene synthesis (1-aminocyclopropane-1-carboxylic acid synthase and 1-aminocyclopropane-1-carboxylic acid oxidase), color (phytoene synthase), firmness (polygalacturonase), and sugar accumulation (acid invertase). The expression of ripening-specific genes was affected by CO2 regardless of whether their induction was ethylene- or development-dependent. It is proposed that the inhibition of tomato fruit ripening by high CO2 is due, in part, to the suppression of the expression of ripening-associated genes, which is probably related to the stress effect exerted by high CO2. PMID:12223703

  11. Titanate nanotube thin films with enhanced thermal stability and high-transparency prepared from additive-free sols

    NASA Astrophysics Data System (ADS)

    Kőrösi, László; Papp, Szilvia; Hornok, Viktória; Oszkó, Albert; Petrik, Péter; Patko, Daniel; Horvath, Robert; Dékány, Imre

    2012-08-01

    Titanate nanotubes were synthesized from TiO2 in alkaline medium by a conventional hydrothermal method (150 °C, 4.7 bar). To obtain hydrogen titanates, the as-prepared sodium titanates were treated with either HCl or H3PO4 aqueous solutions. A simple synthesis procedure was devised for stable titanate nanotube sols without using any additives. These highly stable ethanolic sols can readily be used to prepare transparent titanate nanotube thin films of high quality. The resulting samples were studied by X-ray diffraction, N2-sorption measurements, Raman spectroscopy, transmission and scanning electron microscopy, X-ray photoelectron spectroscopy and spectroscopic ellipsometry. The comparative results of using two kinds of acids shed light on the superior thermal stability of the H3PO4-treated titanate nanotubes (P-TNTs). X-ray photoelectron spectroscopy revealed that P-TNTs contains P in the near-surface region and the thermal stability was enhanced even at a low (˜0.5 at%) concentration of P. After calcination at 500 °C, the specific surface areas of the HCl- and H3PO4-treated samples were 153 and 244 m2 g-1, respectively. The effects of H3PO4 treatment on the structure, morphology and porosity of titanate nanotubes are discussed.

  12. Additive Manufacturing of a Photo-Cross-Linkable Polymer via Direct Melt Electrospinning Writing for Producing High Strength Structures.

    PubMed

    Chen, Fei; Hochleitner, Gernot; Woodfield, Tim; Groll, Juergen; Dalton, Paul D; Amsden, Brian G

    2016-01-11

    Melt electrospinning writing (MEW) is an emerging additive manufacturing technique that enables the design and fabrication of micrometer-thin fibrous scaffolds made of biocompatible and biodegradable polymers. By using a computer-aided deposition process, a unique control over pore size and interconnectivity of the resulting scaffolds is achieved, features highly interesting for tissue engineering applications. However, MEW has been mainly used to process low melting point thermoplastics such as poly(ε-caprolactone). Since this polymer exhibits creep and a reduction in modulus upon hydration, we manufactured scaffolds of poly(L-lactide-co-ε-caprolactone-co-acryloyl carbonate) (poly(LLA-ε-CL-AC)), a photo-cross-linkable and biodegradable polymer, for the first time. We show that the stiffness of the scaffolds increases significantly (up to ∼10-fold) after cross-linking by UV irradiation at room temperature, compared with un-cross-linked microfiber scaffolds. The preservation of stiffness and high average fiber modulus (370 ± 166 MPa) within the cross-linked hydrated scaffolds upon repetitive loading (10% strain at 1 Hz up to 200,000 cycles) suggests that the prepared scaffolds may be of potential interest for soft connective tissue engineering applications. Moreover, the approach can be readily adapted through manipulation of polymer properties and scaffold geometry to prepare structures with mechanical properties suitable for other tissue engineering applications.

  13. Optimal welding parameters for very high power ultrasonic additive manufacturing of smart structures with aluminum 6061 matrix

    NASA Astrophysics Data System (ADS)

    Wolcott, Paul J.; Hehr, Adam; Dapino, Marcelo J.

    2014-03-01

    Ultrasonic additive manufacturing (UAM) is a recent solid state manufacturing process that combines ad- ditive joining of thin metal tapes with subtractive milling operations to generate near net shape metallic parts. Due to the minimal heating during the process, UAM is a proven method of embedding Ni-Ti, Fe-Ga, and PVDF to create active metal matrix composites. Recently, advances in the UAM process utilizing 9 kW very high power (VHP) welding has improved bonding properties, enabling joining of high strength materials previously unweldable with 1 kW low power UAM. Consequently, a design of experiments study was conducted to optimize welding conditions for aluminum 6061 components. This understanding is critical in the design of UAM parts containing smart materials. Build parameters, including weld force, weld speed, amplitude, and temperature were varied based on a Taguchi experimental design matrix and tested for me- chanical strength. Optimal weld parameters were identi ed with statistical methods including a generalized linear model for analysis of variance (ANOVA), mean e ects plots, and interaction e ects plots.

  14. Additive Manufacturing of a Photo-Cross-Linkable Polymer via Direct Melt Electrospinning Writing for Producing High Strength Structures.

    PubMed

    Chen, Fei; Hochleitner, Gernot; Woodfield, Tim; Groll, Juergen; Dalton, Paul D; Amsden, Brian G

    2016-01-11

    Melt electrospinning writing (MEW) is an emerging additive manufacturing technique that enables the design and fabrication of micrometer-thin fibrous scaffolds made of biocompatible and biodegradable polymers. By using a computer-aided deposition process, a unique control over pore size and interconnectivity of the resulting scaffolds is achieved, features highly interesting for tissue engineering applications. However, MEW has been mainly used to process low melting point thermoplastics such as poly(ε-caprolactone). Since this polymer exhibits creep and a reduction in modulus upon hydration, we manufactured scaffolds of poly(L-lactide-co-ε-caprolactone-co-acryloyl carbonate) (poly(LLA-ε-CL-AC)), a photo-cross-linkable and biodegradable polymer, for the first time. We show that the stiffness of the scaffolds increases significantly (up to ∼10-fold) after cross-linking by UV irradiation at room temperature, compared with un-cross-linked microfiber scaffolds. The preservation of stiffness and high average fiber modulus (370 ± 166 MPa) within the cross-linked hydrated scaffolds upon repetitive loading (10% strain at 1 Hz up to 200,000 cycles) suggests that the prepared scaffolds may be of potential interest for soft connective tissue engineering applications. Moreover, the approach can be readily adapted through manipulation of polymer properties and scaffold geometry to prepare structures with mechanical properties suitable for other tissue engineering applications. PMID:26620885

  15. Endosialin‑expressing bone sarcoma stem‑like cells are highly tumor‑initiating and invasive.

    PubMed

    Sun, Dong-Xiu; Liao, Guang-Jun; Liu, Ke-Gui; Jian, Han

    2015-10-01

    It has been reported that the presence of a small group of cancer stem‑like 'side population (SP)' cells is responsible for therapy failure and tumor recurrence. The present study demonstrated that primary human osteosarcoma samples contained a SP of about 3.9% which overexpressed ABC transporters, including ABCA1, ABCB1, ABCB2 and ABCG2, which are associated with drug resistance and may have contributed to multi‑drug resistance of SP cells. Furthermore, these SP cells displayed increased expression of endosialin (CD248) and other stem cell surface proteins, including CD133, octamer‑binding transcription factor 3/4A, Nanog and Nestin, which are ultimately responsible for high self‑renewal and deregulated cell proliferation. In addition, it was shown that endosialin‑overexpressing SP cells were able to regenerate the tumor population and had a high invasive potential. Therefore, the present study suggested that osteosarcoma SP cells were cancer stem cells, as they displayed stem‑like properties; furthermore, endosialin may be a potential target to prevent osteosarcoma recurrence following chemotherapy. PMID:26300407

  16. High-level expression and characterization of a novel serine protease in Pichia pastoris by multi-copy integration.

    PubMed

    Shu, Min; Shen, Wei; Yang, Shihui; Wang, Xiaojuan; Wang, Fei; Wang, Yaping; Ma, Lixin

    2016-10-01

    A novel serine protease from Trichoderma koningii (SPTK) was synthesized and expressed in Pichia pastoris. The recombinant SPTK was completely inhibited by phenyl methyl sulfonyl fluoride (PMSF), suggesting that SPTK belonged to the subgroup of serine proteases. The optimum pH and temperature for the recombinant SPTK reaction were 6.0 and 55°C, respectively. SPTK performed a tolerance to most organic solvents and metal ions, and the addition of Triton X-100 exhibited an activation of SPTK up to 243% of its initial activity but SDS strongly inhibited. Moreover, our study showed that a portion of SPTK was N-glycosylated during fermentation. The activity and thermal stability of the recombinant SPTK were improved after the removal of glycosylation, and the N-glycosylation of SPTK could be efficiently removed through co-culture with P. pastoris strains expressing Endo-β-N-acetylglucosaminidase H. We constructed expression vectors harboring from one to four repeats of Sptk-expressing cassettes via an in vitro BioBrick assembly approach. And the result of quantitative polymerase chain reaction (qPCR) indicated that the tandem expression cassettes were integrated into the genome of P. pastoris through a single recombination event. These strains were used to study the correlation between the gene copy number and the expression level of SPTK. The results of qPCR and enzyme activity assays indicated that the copy number variation of Sptk gene generally had a positive effect on the expression level of SPTK, while an increase in integration of target gene did not guarantee its high expression. The maximum yield and specific activity of SPTK in P. pastoris were obtained from the recombinant yeast strain harboring two-copy tandem Sptk-expressing cassettes, the yield reached 0.48g/l after a 6-d induction using menthol in shake flasks and 3.2g/l in high-density fermentation with specific activity of 5200U/mg. In addition, the recombinant SPTK could efficiently degrade chicken

  17. High-level expression and characterization of a novel serine protease in Pichia pastoris by multi-copy integration.

    PubMed

    Shu, Min; Shen, Wei; Yang, Shihui; Wang, Xiaojuan; Wang, Fei; Wang, Yaping; Ma, Lixin

    2016-10-01

    A novel serine protease from Trichoderma koningii (SPTK) was synthesized and expressed in Pichia pastoris. The recombinant SPTK was completely inhibited by phenyl methyl sulfonyl fluoride (PMSF), suggesting that SPTK belonged to the subgroup of serine proteases. The optimum pH and temperature for the recombinant SPTK reaction were 6.0 and 55°C, respectively. SPTK performed a tolerance to most organic solvents and metal ions, and the addition of Triton X-100 exhibited an activation of SPTK up to 243% of its initial activity but SDS strongly inhibited. Moreover, our study showed that a portion of SPTK was N-glycosylated during fermentation. The activity and thermal stability of the recombinant SPTK were improved after the removal of glycosylation, and the N-glycosylation of SPTK could be efficiently removed through co-culture with P. pastoris strains expressing Endo-β-N-acetylglucosaminidase H. We constructed expression vectors harboring from one to four repeats of Sptk-expressing cassettes via an in vitro BioBrick assembly approach. And the result of quantitative polymerase chain reaction (qPCR) indicated that the tandem expression cassettes were integrated into the genome of P. pastoris through a single recombination event. These strains were used to study the correlation between the gene copy number and the expression level of SPTK. The results of qPCR and enzyme activity assays indicated that the copy number variation of Sptk gene generally had a positive effect on the expression level of SPTK, while an increase in integration of target gene did not guarantee its high expression. The maximum yield and specific activity of SPTK in P. pastoris were obtained from the recombinant yeast strain harboring two-copy tandem Sptk-expressing cassettes, the yield reached 0.48g/l after a 6-d induction using menthol in shake flasks and 3.2g/l in high-density fermentation with specific activity of 5200U/mg. In addition, the recombinant SPTK could efficiently degrade chicken

  18. A Novel Dual Expression Platform for High Throughput Functional Screening of Phage Libraries in Product like Format

    PubMed Central

    Mugabe, Sheila; Gao, Changshou; Tkaczyk, Christine; Mazor, Yariv; Pavlik, Peter; Wu, Herren; Dall’Acqua, William; Chowdhury, Partha Sarathi

    2015-01-01

    High throughput screenings of single chain Fv (scFv) antibody phage display libraries are currently done as soluble scFvs produced in E.coli. Due to endotoxin contaminations from bacterial cells these preparations cannot be reliably used in mammalian cell based assays. The monovalent nature and lack of Fc in soluble scFvs prevent functional assays that are dependent on target cross linking and/or Fc functions. A convenient approach is to convert scFvs into scFv.Fc fusion proteins and express them in mammalian cell lines for screening. This approach is low throughput and is only taken after primary screening of monovalent scFvs that are expressed in bacteria. There is no platform at present that combines the benefits of both bacterial and mammalian expression system for screening phage library output. We have, therefore, developed a novel dual expression vector, called pSplice, which can be used to express scFv.Fc fusion proteins both in E.coli and mammalian cell lines. The hallmark of the vector is an engineered intron which houses the bacterial promoter and signal peptide for expression and secretion of scFv.Fc in E.coli. When the vector is transfected into a mammalian cell line, the intron is efficiently spliced out resulting in a functional operon for expression and secretion of the scFv.Fc fusion protein into the culture medium. By applying basic knowledge of mammalian introns and splisosome, we designed this vector to enable screening of phage libraries in a product like format. Like IgG, the scFv.Fc fusion protein is bi-valent for the antigen and possesses Fc effector functions. Expression in E.coli maintains the speed of the bacterial expression platform and is used to triage clones based on binding and other assays that are not sensitive to endotoxin. Triaged clones are then expressed in a mammalian cell line without the need for any additional cloning steps. Conditioned media from the mammalian cell line containing the fusion proteins are then used for

  19. A Novel Dual Expression Platform for High Throughput Functional Screening of Phage Libraries in Product like Format.

    PubMed

    Xiao, Xiaodong; Chen, Yan; Mugabe, Sheila; Gao, Changshou; Tkaczyk, Christine; Mazor, Yariv; Pavlik, Peter; Wu, Herren; Dall'Acqua, William; Chowdhury, Partha Sarathi

    2015-01-01

    High throughput screenings of single chain Fv (scFv) antibody phage display libraries are currently done as soluble scFvs produced in E.coli. Due to endotoxin contaminations from bacterial cells these preparations cannot be reliably used in mammalian cell based assays. The monovalent nature and lack of Fc in soluble scFvs prevent functional assays that are dependent on target cross linking and/or Fc functions. A convenient approach is to convert scFvs into scFv.Fc fusion proteins and express them in mammalian cell lines for screening. This approach is low throughput and is only taken after primary screening of monovalent scFvs that are expressed in bacteria. There is no platform at present that combines the benefits of both bacterial and mammalian expression system for screening phage library output. We have, therefore, developed a novel dual expression vector, called pSplice, which can be used to express scFv.Fc fusion proteins both in E.coli and mammalian cell lines. The hallmark of the vector is an engineered intron which houses the bacterial promoter and signal peptide for expression and secretion of scFv.Fc in E.coli. When the vector is transfected into a mammalian cell line, the intron is efficiently spliced out resulting in a functional operon for expression and secretion of the scFv.Fc fusion protein into the culture medium. By applying basic knowledge of mammalian introns and splisosome, we designed this vector to enable screening of phage libraries in a product like format. Like IgG, the scFv.Fc fusion protein is bi-valent for the antigen and possesses Fc effector functions. Expression in E.coli maintains the speed of the bacterial expression platform and is used to triage clones based on binding and other assays that are not sensitive to endotoxin. Triaged clones are then expressed in a mammalian cell line without the need for any additional cloning steps. Conditioned media from the mammalian cell line containing the fusion proteins are then used for

  20. Genome of turbot rhabdovirus exhibits unusual non-coding regions and an additional ORF that could be expressed in fish cell.

    PubMed

    Zhu, Ruo-Lin; Lei, Xiao-Ying; Ke, Fei; Yuan, Xiu-Ping; Zhang, Qi-Ya

    2011-02-01

    Genomic sequence of Scophthalmus maximus rhabdovirus (SMRV) isolated from diseased turbot has been characterized. The complete genome of SMRV comprises 11,492 nucleotides and encodes five typical rhabdovirus genes N, P, M, G and L. In addition, two open reading frames (ORF) are predicted overlapping with P gene, one upstream of P and smaller than P (temporarily called Ps), and another in P gene which may encodes a protein similar to the vesicular stomatitis virus C protein. The C ORF is contained within the P ORF. The five typical proteins share the highest sequence identities (48.9%) with the corresponding proteins of rhabdoviruses in genus Vesiculovirus. Phylogenetic analysis of partial L protein sequence indicates that SMRV is close to genus Vesiculovirus. The first 13 nucleotides at the ends of the SMRV genome are absolutely inverse complementarity. The gene junctions between the five genes show conserved polyadenylation signal (CATGA(7)) and intergenic dinucleotide (CT) followed by putative transcription initiation sequence A(A/G)(C/G)A(A/G/T), which are different from known rhabdoviruses. The entire Ps ORF was cloned and expressed, and used to generate polyclonal antibody in mice. One obvious band could be detected in SMRV-infected carp leucocyte cells (CLCs) by anti-Ps/C serum via Western blot, and the subcellular localization of Ps-GFP fusion protein exhibited cytoplasm distribution as multiple punctuate or doughnut shaped foci of uneven size.

  1. Titanate nanotube thin films with enhanced thermal stability and high-transparency prepared from additive-free sols

    SciTech Connect

    Koroesi, Laszlo; Papp, Szilvia; Hornok, Viktoria; Oszko, Albert; Petrik, Peter; Patko, Daniel; Horvath, Robert; Dekany, Imre

    2012-08-15

    Titanate nanotubes were synthesized from TiO{sub 2} in alkaline medium by a conventional hydrothermal method (150 Degree-Sign C, 4.7 bar). To obtain hydrogen titanates, the as-prepared sodium titanates were treated with either HCl or H{sub 3}PO{sub 4} aqueous solutions. A simple synthesis procedure was devised for stable titanate nanotube sols without using any additives. These highly stable ethanolic sols can readily be used to prepare transparent titanate nanotube thin films of high quality. The resulting samples were studied by X-ray diffraction, N{sub 2}-sorption measurements, Raman spectroscopy, transmission and scanning electron microscopy, X-ray photoelectron spectroscopy and spectroscopic ellipsometry. The comparative results of using two kinds of acids shed light on the superior thermal stability of the H{sub 3}PO{sub 4}-treated titanate nanotubes (P-TNTs). X-ray photoelectron spectroscopy revealed that P-TNTs contains P in the near-surface region and the thermal stability was enhanced even at a low ({approx}0.5 at%) concentration of P. After calcination at 500 Degree-Sign C, the specific surface areas of the HCl- and H{sub 3}PO{sub 4}-treated samples were 153 and 244 m{sup 2} g{sup -1}, respectively. The effects of H{sub 3}PO{sub 4} treatment on the structure, morphology and porosity of titanate nanotubes are discussed. - Graphical Abstract: TEM picture (left) shows P-TNTs with diameters about 5-6 nm. Inset shows a stable titanate nanotube sol illuminated by a 532 nm laser beam. Due to the presence of the nanoparticles the way of the light is visible in the sol. Cross sectional SEM picture (right) as well as ellipsometry revealed the formation of optical quality P-TNT films with thicknesses below 50 nm. Highlights: Black-Right-Pointing-Pointer H{sub 3}PO{sub 4} treatment led to TNTs with high surface area even after calcination at 500 Degree-Sign C. Black-Right-Pointing-Pointer H{sub 3}PO{sub 4}-treated TNTs preserved their nanotube morphology up to 500

  2. EFFECTS OF QUARTZ PARTICLE SIZE AND SUCROSE ADDITION ON MELTING BEHAVIOR OF A MELTER FEED FOR HIGH-LEVEL GLASS

    SciTech Connect

    MARCIAL J; KRUGER AA; HRMA PR; SCHWEIGER MJ; SWEARINGEN KJ; TEGROTENHUIS WE; HENAGER SH

    2010-07-28

    The behavior of melter feed (a mixture of nuclear waste and glass-forming additives) during waste-glass processing has a significant impact on the rate of the vitrification process. We studied the effects of silica particle size and sucrose addition on the volumetric expansion (foaming) of a high-alumina feed and the rate of dissolution of silica particles in feed samples heated at 5 C/min up to 1200 C. The initial size of quartz particles in feed ranged from 5 to 195 {micro}m. The fraction of the sucrose added ranged from 0 to 0.20 g per g glass. Extensive foaming occurred only in feeds with 5-{micro}m quartz particles; particles {ge}150 {micro}m formed clusters. Particles of 5 {micro}m completely dissolved by 900 C whereas particles {ge}150 {micro}m did not fully dissolve even when the temperature reached 1200 C. Sucrose addition had virtually zero impact on both foaming and the dissolution of silica particles. Over 100 sites in the United States are currently tasked with the storage of nuclear waste. The largest is the Hanford Site located in southeastern Washington State with 177 subterranean tanks containing over fifty-million gallons of nuclear waste from plutonium production from 1944 through 1987. This waste will be vitrified at the Hanford Tank Waste Treatment and Immobilization Plant. In the vitrification process, feed is charged into a melter and converted into glass to be ultimately stored in a permanent repository. The duration of waste-site cleanups by the vitrification process depends on the rate of melting, i.e., on the rate of the feed-to-glass conversion. Foaming associated with the melting process and the rate of dissolution of quartz particles (silica being the major glass-forming additive) are assumed to be important factors that influence the rate of melting. Previous studies on foaming of high-alumina feed demonstrated that varying the makeup of a melter feed has a significant impact on foaming. The volume of feeds that contained 5-{micro

  3. pUNISHER: a high-level expression cassette for use with recombinant viral vectors for rapid and long term in vivo neuronal expression in the CNS.

    PubMed

    Montesinos, Monica S; Chen, Zuxin; Young, Samuel M

    2011-12-01

    Fast onset and high-level neurospecific transgene expression in vivo is of importance for many areas in neuroscience, from basic to translational, and can significantly reduce the amount of vector load required to maintain transgene expression in vivo. In this study, we tested various cis elements to optimize transgene expression at transcriptional, posttranscriptional, and posttranslational levels and combined them together to create the high-level neuronal transgene expression cassette pUNISHER. Using a second-generation adenoviral vector system in combination with the pUNISHER cassette, we characterized its rate of onset of detectable expression and levels of expression compared with a neurospecific expression cassette driven by the 470-bp human synapsin promoter in vitro and in vivo. Our results demonstrate in primary neurons that the pUNISHER cassette, in a recombinant adenovirus type 5 background, led to a faster rate of onset of detectable transgene expression and higher level of transgene expression. More importantly, this cassette led to highly correlated neuronal expression in vivo and to stable transgene expression up to 30 days in the auditory brain stem with no toxicity on the characteristics of synaptic transmission and plasticity at the calyx of Held synapse. Thus the pUNISHER cassette is an ideal high-level neuronal expression cassette for use in vivo for neuroscience applications. PMID:21957229

  4. Unfolding and refolding of Coprinus cinereus peroxidase at high pH, in urea, and at high temperature. Effect of organic and ionic additives on these processes.

    PubMed

    Tams, J W; Welinder, K G

    1996-06-11

    The unfolding and refolding rates of the heme-and Ca2+ -containing Coprinus cinereus peroxidase (CIP) have been measured at pH 12.1, in 4 M urea, and at 61.2 degrees C. The change in peroxidase activity paralleled the change in the Soret band absorbance of the heme group. The unfolding rate constant (ku), was determined independently in thermolysin digestion and EDTA experiments at 59.4 degrees C. Both gave ku values of 1.5 ms-1, and also showed that the presence of 4 mM EDTA made CIP unfolding practically irreversible. Unfolding and refolding rates could therefore be determined under identical conditions of denaturation having either EDTA or Ca2+ in excess. The refolding rates at high pH and in 4 M urea were measured by adding Ca2+ to the unfolded CIP, whereas refolding at 61.2 degrees C was evaluated by comparing the unfolding carried out under reversible (excess of Ca2+) and irreversible conditions (excess EDTA). The activation energies for the unfolding at 61.2 degrees C are approximately delta G++(u) 100, T delta S++(u) 200, and delta H++(u) 300 kJ/mol. Five different additives, glycerol, EtOH, Na2SO4, guanidinium chloride (GdmCl), and NaCl, all at 100 mM, were used as probes to evaluate the mechanism of base, urea, and heat on unfolding and refolding. Salts destabilized CIP at high pH, primarily by enhancing the unfolding rate but also by decreasing the refolding rate. Glycerol had the reverse effects and thus stabilized CIP at high pH. The unfolding rate in urea was only slightly affected by the additives, with the exception of GdmCl which enhanced the unfolding rate. The refolding rate was decreased in urea by EtOH and GdmCl, in contrast to glycerol and Na2SO4 which increased the refolding rate. At high temperature the unfolding was affected only slightly by the additives, except for GdmCl, and to a lesser extent NaCl, which enhanced the unfolding rate. The refolding rates were greatly decreased by Na2SO4, EtOH, and GdmCl, whereas glycerol and Nacl enhanced

  5. Express

    Integrated Risk Information System (IRIS)

    Express ; CASRN 101200 - 48 - 0 Human health assessment information on a chemical substance is included in the IRIS database only after a comprehensive review of toxicity data , as outlined in the IRIS assessment development process . Sections I ( Health Hazard Assessments for Noncarcinogenic Effect

  6. High-throughput sequencing-based genome-wide identification of microRNAs expressed in developing cotton seeds.

    PubMed

    Wang, YanMei; Ding, Yan; Yu, DingWei; Xue, Wei; Liu, JinYuan

    2015-08-01

    MicroRNAs (miRNAs) have been shown to play critical regulatory roles in gene expression in cotton. Although a large number of miRNAs have been identified in cotton fibers, the functions of miRNAs in seed development remain unexplored. In this study, a small RNA library was constructed from cotton seeds sampled at 15 days post-anthesis (DPA) and was subjected to high-throughput sequencing. A total of 95 known miRNAs were detected to be expressed in cotton seeds. The expression pattern of these identified miRNAs was profiled and 48 known miRNAs were differentially expressed between cotton seeds and fibers at 15 DPA. In addition, 23 novel miRNA candidates were identified in 15-DPA seeds. Putative targets for 21 novel and 87 known miRNAs were successfully predicted and 900 expressed sequence tag (EST) sequences were proposed to be candidate target genes, which are involved in various metabolic and biological processes, suggesting a complex regulatory network in developing cotton seeds. Furthermore, miRNA-mediated cleavage of three important transcripts in vivo was validated by RLM-5' RACE. This study is the first to show the regulatory network of miRNAs that are involved in developing cotton seeds and provides a foundation for future studies on the specific functions of these miRNAs in seed development.

  7. Identifying reproducible cancer-associated highly expressed genes with important functional significances using multiple datasets

    PubMed Central

    Huang, Haiyan; Li, Xiangyu; Guo, You; Zhang, Yuncong; Deng, Xusheng; Chen, Lufei; Zhang, Jiahui; Guo, Zheng; Ao, Lu

    2016-01-01

    Identifying differentially expressed (DE) genes between cancer and normal tissues is of basic importance for studying cancer mechanisms. However, current methods, such as the commonly used Significance Analysis of Microarrays (SAM), are biased to genes with low expression levels. Recently, we proposed an algorithm, named the pairwise difference (PD) algorithm, to identify highly expressed DE genes based on reproducibility evaluation of top-ranked expression differences between paired technical replicates of cells under two experimental conditions. In this study, we extended the application of the algorithm to the identification of DE genes between two types of tissue samples (biological replicates) based on several independent datasets or sub-datasets of a dataset, by constructing multiple paired average gene expression profiles for the two types of samples. Using multiple datasets for lung and esophageal cancers, we demonstrated that PD could identify many DE genes highly expressed in both cancer and normal tissues that tended to be missed by the commonly used SAM. These highly expressed DE genes, including many housekeeping genes, were significantly enriched in many conservative pathways, such as ribosome, proteasome, phagosome and TNF signaling pathways with important functional significances in oncogenesis. PMID:27796338

  8. Cloning, expression and biochemical characterization of one Epsilon-class (GST-3) and ten Delta-class (GST-1) glutathione S-transferases from Drosophila melanogaster, and identification of additional nine members of the Epsilon class.

    PubMed

    Sawicki, Rafał; Singh, Sharda P; Mondal, Ashis K; Benes, Helen; Zimniak, Piotr

    2003-03-01

    From the fruitfly, Drosophila melanogaster, ten members of the cluster of Delta-class glutathione S-transferases (GSTs; formerly denoted as Class I GSTs) and one member of the Epsilon-class cluster (formerly GST-3) have been cloned, expressed in Escherichia coli, and their catalytic properties have been determined. In addition, nine more members of the Epsilon cluster have been identified through bioinformatic analysis but not further characterized. Of the 11 expressed enzymes, seven accepted the lipid peroxidation product 4-hydroxynonenal as substrate, and nine were active in glutathione conjugation of 1-chloro-2,4-dinitrobenzene. Since the enzymically active proteins included the gene products of DmGSTD3 and DmGSTD7 which were previously deemed to be pseudogenes, we investigated them further and determined that both genes are transcribed in Drosophila. Thus our present results indicate that DmGSTD3 and DmGSTD7 are probably functional genes. The existence and multiplicity of insect GSTs capable of conjugating 4-hydroxynonenal, in some cases with catalytic efficiencies approaching those of mammalian GSTs highly specialized for this function, indicates that metabolism of products of lipid peroxidation is a highly conserved biochemical pathway with probable detoxification as well as regulatory functions.

  9. Improving Co-Amorphous Drug Formulations by the Addition of the Highly Water Soluble Amino Acid, Proline

    PubMed Central

    Jensen, Katrine Tarp; Löbmann, Korbinian; Rades, Thomas; Grohganz, Holger

    2014-01-01

    Co-amorphous drug amino acid mixtures were previously shown to be a promising approach to create physically stable amorphous systems with the improved dissolution properties of poorly water-soluble drugs. The aim of this work was to expand the co-amorphous drug amino acid mixture approach by combining the model drug, naproxen (NAP), with an amino acid to physically stabilize the co-amorphous system (tryptophan, TRP, or arginine, ARG) and a second highly soluble amino acid (proline, PRO) for an additional improvement of the dissolution rate. Co-amorphous drug-amino acid blends were prepared by ball milling and investigated for solid state characteristics, stability and the dissolution rate enhancement of NAP. All co-amorphous mixtures were stable at room temperature and 40 °C for a minimum of 84 days. PRO acted as a stabilizer for the co-amorphous system, including NAP–TRP, through enhancing the molecular interactions in the form of hydrogen bonds between all three components in the mixture. A salt formation between the acidic drug, NAP, and the basic amino acid, ARG, was found in co-amorphous NAP–ARG. In comparison to crystalline NAP, binary NAP–TRP and NAP–ARG, it could be shown that the highly soluble amino acid, PRO, improved the dissolution rate of NAP from the ternary co-amorphous systems in combination with either TRP or ARG. In conclusion, both the solubility of the amino acid and potential interactions between the molecules are critical parameters to consider in the development of co-amorphous formulations. PMID:25025400

  10. Meta-analysis of high-latitude nitrogen-addition and warming studies imply ecological mechanisms overlooked by land models

    DOE PAGES

    Bouskill, N. J.; Riley, W. J.; Tang, J.

    2014-08-18

    Accurate representation of ecosystem processes in land models is crucial for reducing predictive uncertainty in energy and greenhouse gas feedbacks with the atmosphere. Here we describe an observational and modeling meta-analysis approach to benchmark land models, and apply the method to the land model CLM4.5 with two versions of belowground biogeochemistry. We focused our analysis on the above and belowground high-latitude ecosystem responses to warming and nitrogen addition, and identified mechanisms absent, or poorly parameterized in CLM4.5. While the two model versions predicted similar trajectories for soil carbon stocks following both types of perturbation, other variables (e.g., belowground respiration) differedmore » from the observations in both magnitude and direction, indicating the underlying mechanisms are inadequate for representing high-latitude ecosystems. The observational synthesis attribute these differences to missing representations of microbial dynamics, characterization of above and belowground functional processes, and nutrient competition. We use the observational meta-analyses to discuss potential approaches to improving the current models (e.g., the inclusion of dynamic vegetation or different microbial functional guilds), however, we also raise a cautionary note on the selection of data sets and experiments to be included in a meta-analysis. For example, the concentrations of nitrogen applied in the synthesized field experiments (average =72 kg ha-1 yr-1) are many times higher than projected soil nitrogen concentrations (from nitrogen deposition and release during mineralization), which preclude a rigorous evaluation of the model responses to nitrogen perturbation. Overall, we demonstrate here that elucidating ecological mechanisms via meta-analysis can identify deficiencies in both ecosystem models and empirical experiments.« less

  11. 4-(Trifluoromethyl)-benzonitrile: A novel electrolyte additive for lithium nickel manganese oxide cathode of high voltage lithium ion battery

    NASA Astrophysics Data System (ADS)

    Huang, Wenna; Xing, Lidan; Wang, Yating; Xu, Mengqing; Li, Weishan; Xie, Fengchao; Xia, Shengan

    2014-12-01

    In this work, 4-(Trifluoromethyl)-benzonitrile (4-TB) is used as a novel electrolyte additive for LiNi0.5Mn1.5O4 cathode of high voltage lithium ion battery. Charge-discharge tests show that the cyclic stability of LiNi0.5Mn1.5O4 is significantly improved by using 0.5 wt.% 4-TB. With using 4-TB, LiNi0.5Mn1.5O4 delivers an initial capacity of 133 mAh g-1 and maintains 121 mAh g-1 after 300 cycles with a capacity retention of 91%, compared to the 75% of that using base electrolyte (1 M LiPF6 in ethylene carbonate(EC)/dimethyl carbonate(DMC)). The results from linear sweep voltammetry, density functional theory calculations, electrochemical impedance spectroscopy, scanning electron microscope, energy dispersive spectroscopy, Fourier transform infrared, and inductively coupled plasma, indicate that 4-TB has lower oxidative stability than EC and DMC, and is preferentially oxidized on LiNi0.5Mn1.5O4 forming a low-impedance protective film, which prevents the subsequent oxidation decomposition of the electrolyte and suppresses the manganese dissolution from LiNi0.5Mn1.5O4.

  12. Effect of addition of organic waste on reduction of Escherichia coli during cattle feces composting under high-moisture condition.

    PubMed

    Hanajima, Dai; Kuroda, Kazutaka; Fukumoto, Yasuyuki; Haga, Kiyonori

    2006-09-01

    To ensure Escherichia coli reduction during cattle feces composting, co-composting with a variety of organic wastes was examined. A mixture of dairy cattle feces and shredded rice straw (control) was blended with organic wastes (tofu residue, rice bran, rapeseed meal, dried chicken feces, raw chicken feces, or garbage), and composted using a bench-scale composter under the high-moisture condition (78%). The addition of organic waste except chicken feces brought about maximum temperatures of more than 55 degrees C and significantly reduced the number of E. coli from 10(6) to below 10(2)CFU/g-wet after seven days composting, while in the control treatment, E. coli survived at the same level as that of raw feces. Enhancements of the thermophilic phase and E. coli reduction were related to the initial amount of easily digestible carbon in mass determined as BOD. BOD value more than 166.2 mg O2/DMg brought about significant E. coli reduction.

  13. High-yield soluble expression of recombinant influenza virus antigens from Escherichia coli and their potential uses in diagnosis.

    PubMed

    Jang, Yo Han; Cho, Seung Hee; Son, Ahyun; Lee, Yun Ha; Lee, Jinhee; Lee, Kwang-Hee; Seong, Baik Lin

    2014-02-01

    Although antiviral drugs and vaccines have been successful for mitigating influenza virus infections, the lack of general technical platform for the timely supply of soluble and highly purified influenza viral antigens presents a serious bottleneck for the subsequent analysis for the effective control of the viral disease. Using the Escherichia coli (E. coli) lysyl tRNA synthetase (LysRS) as a novel fusion partner, this study reports the soluble expression of influenza viral proteins in E. coli host, construction of antibody library against the virus, and detection of anti-influenza antibodies using the expressed viral antigens. When influenza A and B viral proteins were fused with the LysRS, the fusion proteins were expressed predominantly as soluble forms and their production yields were high enough to be amenable to immunization protocols in rabbits for antibody generation. The produced antibodies showed high level binding specificity against the respective viral proteins, with cross-reactivity across heterologous viruses within the same type of influenza viruses. In addition, LysRS-HA fusion protein could bind specifically to anti-HA antibodies in the virus-infected mouse serum in widely accepted two detection methods, Western blot and ELISA. These results present a convenient tool for the production of antibodies specific to the virus as well as the rapid detection of influenza viral infections, ultimately contributing to the control of influenza viruses including highly pathogenic H5N1, pandemic H1N1, or the recent H7N9 influenza viruses.

  14. High Laccase Expression by Trametes versicolor in a Simulated Textile Effluent with Different Carbon Sources and PHs.

    PubMed

    Ottoni, Cristiane; Simões, Marta F; Fernandes, Sara; Santos, Cledir R; Lima, Nelson

    2016-01-01

    Textile effluents are highly polluting and have variable and complex compositions. They can be extremely complex, with high salt concentrations and alkaline pHs. A fixed-bed bioreactor was used in the present study to simulate a textile effluent treatment, where the white-rot fungus, Trametes versicolor, efficiently decolourised the azo dye Reactive Black 5 over 28 days. This occurred under high alkaline conditions, which is unusual, but advantageous, for successful decolourisation processes. Active dye decolourisation was maintained by operation in continuous culture. Colour was eliminated during the course of operation and maximum laccase (Lcc) activity (80.2 U∙L(-1)) was detected after glycerol addition to the bioreactor. Lcc2 gene expression was evaluated with different carbon sources and pH values based on reverse transcriptase-PCR (polymerase chain reaction). Glycerol was shown to promote the highest lcc2 expression at pH 5.5, followed by sucrose and then glucose. The highest levels of expression occurred between three and four days, which corroborate the maximum Lcc activity observed for sucrose and glycerol on the bioreactor. These results give new insights into the use of T. versicolor in textile dye wastewater treatment with high pHs. PMID:27490563

  15. High Laccase Expression by Trametes versicolor in a Simulated Textile Effluent with Different Carbon Sources and PHs

    PubMed Central

    Ottoni, Cristiane; Simões, Marta F.; Fernandes, Sara; Santos, Cledir R.; Lima, Nelson

    2016-01-01

    Textile effluents are highly polluting and have variable and complex compositions. They can be extremely complex, with high salt concentrations and alkaline pHs. A fixed-bed bioreactor was used in the present study to simulate a textile effluent treatment, where the white-rot fungus, Trametes versicolor, efficiently decolourised the azo dye Reactive Black 5 over 28 days. This occurred under high alkaline conditions, which is unusual, but advantageous, for successful decolourisation processes. Active dye decolourisation was maintained by operation in continuous culture. Colour was eliminated during the course of operation and maximum laccase (Lcc) activity (80.2 U∙L−1) was detected after glycerol addition to the bioreactor. Lcc2 gene expression was evaluated with different carbon sources and pH values based on reverse transcriptase-PCR (polymerase chain reaction). Glycerol was shown to promote the highest lcc2 expression at pH 5.5, followed by sucrose and then glucose. The highest levels of expression occurred between three and four days, which corroborate the maximum Lcc activity observed for sucrose and glycerol on the bioreactor. These results give new insights into the use of T. versicolor in textile dye wastewater treatment with high pHs. PMID:27490563

  16. High glucose and palmitate increases bone morphogenic protein 4 expression in human endothelial cells

    PubMed Central

    Hong, Oak-Kee; Yoo, Soon-Jib; Son, Jang-Won; Kim, Mee-Kyoung; Baek, Ki-Hyun; Song, Ki-Ho; Cha, Bong-Yun; Jo, Hanjoong

    2016-01-01

    Here, we investigated whether hyperglycemia and/or free fatty acids (palmitate, PAL) aff ect the expression level of bone morphogenic protein 4 (BMP4), a proatherogenic marker, in endothelial cells and the potential role of BMP4 in diabetic vascular complications. To measure BMP4 expression, human umbilical vein endothelial cells (HUVECs) were exposed to high glucose concentrations and/or PAL for 24 or 72 h, and the effects of these treatments on the expression levels of adhesion molecules and reactive oxygen species (ROS) were examined. BMP4 loss-of-function status was achieved via transfection of a BMP4-specific siRNA. High glucose levels increased BMP4 expression in HUVECs in a dose-dependent manner. PAL potentiated such expression. The levels of adhesion molecules and ROS production increased upon treatment with high glucose and/or PAL, but this eff ect was negated when BMP4 was knocked down via siRNA. Signaling of BMP4, a proinflammatory and pro-atherogenic cytokine marker, was increased by hyperglycemia and PAL. BMP4 induced the expression of infl ammatory adhesion molecules and ROS production. Our work suggests that BMP4 plays a role in atherogenesis induced by high glucose levels and/or PAL. PMID:26937213

  17. High glucose and palmitate increases bone morphogenic protein 4 expression in human endothelial cells.

    PubMed

    Hong, Oak-Kee; Yoo, Soon-Jib; Son, Jang-Won; Kim, Mee-Kyoung; Baek, Ki-Hyun; Song, Ki-Ho; Cha, Bong-Yun; Jo, Hanjoong; Kwon, Hyuk-Sang

    2016-03-01

    Here, we investigated whether hyperglycemia and/or free fatty acids (palmitate, PAL) aff ect the expression level of bone morphogenic protein 4 (BMP4), a proatherogenic marker, in endothelial cells and the potential role of BMP4 in diabetic vascular complications. To measure BMP4 expression, human umbilical vein endothelial cells (HUVECs) were exposed to high glucose concentrations and/or PAL for 24 or 72 h, and the effects of these treatments on the expression levels of adhesion molecules and reactive oxygen species (ROS) were examined. BMP4 loss-of-function status was achieved via transfection of a BMP4-specific siRNA. High glucose levels increased BMP4 expression in HUVECs in a dose-dependent manner. PAL potentiated such expression. The levels of adhesion molecules and ROS production increased upon treatment with high glucose and/or PAL, but this eff ect was negated when BMP4 was knocked down via siRNA. Signaling of BMP4, a proinflammatory and pro-atherogenic cytokine marker, was increased by hyperglycemia and PAL. BMP4 induced the expression of infl ammatory adhesion molecules and ROS production. Our work suggests that BMP4 plays a role in atherogenesis induced by high glucose levels and/or PAL. PMID:26937213

  18. Low expression of Bin1, along with high expression of IDO in tumor tissue and draining lymph nodes, are predictors of poor prognosis for esophageal squamous cell cancer patients.

    PubMed

    Jia, Yunlong; Wang, Hongyan; Wang, Yu; Wang, Tingting; Wang, Miao; Ma, Ming; Duan, Yuqing; Meng, Xianli; Liu, Lihua

    2015-09-01

    Indoleamine 2,3-dioxygenase (IDO) has been reported to be involved in esophageal squamous cell cancer (ESCC) progression by promoting immune escape. Previous studies have revealed bridging integrator-1 (Bin1) can inhibit cancer cell growth by suppressing expression of IDO, thus we investigated the correlation between the expression of Bin1 and IDO and their prognostic significances for ESCC patients. Specimens were collected from 196 ESCC patients and detected with flow cytometry, reverse transcription-polymerase chain reaction and immunohistochemistry. We found that in tumor microenvironment (TME) and tumor draining lymph node (TDLN), the proportions of CD3(+) CD4(+) T cell, CD3(+) CD8(+) T cell and CD3(-) CD16(+) CD56(+) NK cell were lower while the proportions of CD3(-) CD19(+) B cell and CD4(+) CD25(+) Treg were higher in specimens with high IDO expression when compared to the specimens with low IDO expression (p < 0.01). In addition, IDO expression was negatively correlated with Bin1 expression at gene and protein level in TME and TDLN. Both the expression of Bin1 and IDO were associated with some clinicopathological parameters including differentiation grade, TNM stage, invasion range, lymph node metastasis (p < 0.05). Moreover, multivariate survival analysis suggested that, along with some other parameters, low expression of Bin1 and high expression of IDO might be independent prognostic factor for ESCC patients. Our results demonstrate that low expression of Bin1, along with high expression of IDO, are predictor for poor prognosis in ESCC and thereby could be used to establish new therapeutic strategies.

  19. High Bak Expression Is Associated with a Favorable Prognosis in Breast Cancer and Sensitizes Breast Cancer Cells to Paclitaxel.

    PubMed

    Luo, Yanwei; Wang, Xinye; Wang, Heran; Xu, Yang; Wen, Qiuyuan; Fan, Songqing; Zhao, Ran; Jiang, Shihe; Yang, Jing; Liu, Yukun; Li, Xiayu; Xiong, Wei; Ma, Jian; Peng, Shuping; Zeng, Zhaoyang; Li, Xiaoling; Phillips, Joshua B; Li, Guiyuan; Tan, Ming; Zhou, Ming

    2015-01-01

    Breast cancer has become the leading cause of cancer-related death among women. A large number of patients become resistant to drug chemotherapy. Paclitaxel (Taxol) is an effective chemotherapeutic agent used to treat cancer patients. Taxol has been widely used in human malignancies including breast cancer because it can stabilize microtubules resulting in cell death by causing an arrest during the G2/M phase of the cell cycle. Pro-apoptotic Bcl-2 antagonist killer 1 (Bak) plays an important role in Taxol-induced apoptosis in breast cancer. In our present study, we investigated the expression of the Bak protein and clinicopathological correlations in a large sample of breast cancer tissues by immunohistochemistry. We found that the percentage of high scores of Bak expression in breast cancer was significantly lower than that of the non-cancerous breast control tissue. In addition, lower Bak expression was positively associated with the clinical TNM stage of breast cancer with a significant decrease in overall survival compared with those with higher Bak expression especially in the Luminal and HER2 subtypes. Importantly, higher Bak expression predicted a favorable clinical outcome in the cases treated with Taxol indicated by a higher overall survival than that of patients with lower Bak expression especially in Luminal and HER2 subtypes. Furthermore, these results were confirmed in vitro since overexpression of Bak sensitized breast cancer cells to Taxol by inhibiting proliferation and promoting apoptosis; in contrast, downregulation of Bak through siRNA transfection inhibited Taxol induced-apoptosis. Therefore, our results demonstrate that Bak acts as a sensitive biomarker and favorable prognostic factor for Taxol treatment in breast cancer. The restoration of Bak expression would be therapeutically beneficial for Taxol resistant breast cancer patients.

  20. SOX11 expression is highly specific for mantle cell lymphoma and identifies the cyclin D1-negative subtype

    PubMed Central

    Mozos, Ana; Royo, Cristina; Hartmann, Elena; De Jong, Daphne; Baró, Cristina; Valera, Alexandra; Fu, Kai; Weisenburger, Dennis D.; Delabie, Jan; Chuang, Shih-Sung; Jaffe, Elaine S.; Ruiz-Marcellan, Carmen; Dave, Sandeep; Rimsza, Lisa; Braziel, Rita; Gascoyne, Randy D.; Solé, Francisco; López-Guillermo, Armando; Colomer, Dolors; Staudt, Louis M.; Rosenwald, Andreas; Ott, German; Jares, Pedro; Campo, Elias

    2009-01-01

    Background Cyclin D1-negative mantle cell lymphoma is difficult to distinguish from other small B-cell lymphomas. The clinical and pathological characteristics of patients with this form of lymphoma have not been well defined. Overexpression of the transcription factor SOX11 has been observed in conventional mantle cell lymphoma. The aim of this study was to determine whether this gene is expressed in cyclin D1-negative mantle cell lymphoma and whether its detection may be useful to identify these tumors. Design and Methods The microarray database of 238 mature B-cell neoplasms was re-examined. SOX11 protein expression was investigated immunohistochemically in 12 cases of cyclin D1-negative mantle cell lymphoma, 54 cases of conventional mantle cell lymphoma, and 209 additional lymphoid neoplasms. Results SOX11 mRNA was highly expressed in conventional and cyclin D1-negative mantle cell lymphoma and in 33% of the cases of Burkitt’s lymphoma but not in any other mature lymphoid neoplasm. SOX11 nuclear protein was detected in 50 cases (93%) of conventional mantle cell lymphoma and also in the 12 cyclin D1-negative cases of mantle cell lymphoma, the six cases of lymphoblastic lymphomas, in two of eight cases of Burkitt’s lymphoma, and in two of three T-prolymphocytic leukemias but was negative in the remaining lymphoid neoplasms. Cyclin D2 and D3 mRNA levels were significantly higher in cyclin D1-negative mantle cell lymphoma than in conventional mantle cell lymphoma but the protein expression was not discriminative. The clinico-pathological features and outcomes of the patients with cyclin D1-negative mantle cell lymphoma identified by SOX11 expression were similar to those of patients with conventional mantle cell lymphoma. Conclusions SOX11 mRNA and nuclear protein expression is a highly specific marker for both cyclin D1-positive and negative mantle cell lymphoma. PMID:19880778

  1. Behavioral effects and CRF expression in brain structures of high- and low-anxiety rats after chronic restraint stress.

    PubMed

    Wisłowska-Stanek, Aleksandra; Lehner, Małgorzata; Skórzewska, Anna; Krząścik, Paweł; Płaźnik, Adam

    2016-09-01

    The aim of our study was to investigate the influence of chronic restraint stress (5 weeks, 3h/day) on behavior and central corticotropin-releasing factor (CRF) expression in rats selected for high (HR) and low anxiety (LR). The conditioned freezing response was used as a discriminating variable. Moreover, we assessed the influence of acute restraint on CRF expression in the brain in HR and LR rats. We found that chronic restraint induced symptoms of anhedonia (decreased consumption of 1% sucrose solution) in HR rats. In addition, HR restraint rats showed an increased learned helplessness behavior (immobility time in the Porsolt test) as well as neophobia in the open field test vs. LR restraint and HR control rats. These behavioral changes were accompanied by a decreased expression of CRF in the paraventricular nucleus of the hypothalamus (pPVN) and the dentate gyrus of the hippocampus (DG) compared to the HR control and LR restraint rat groups, respectively. The acute restraint condition increased the expression of CRF in the pPVN of HR rats compared to the HR control group, and enhanced the expression of CRF in the CA1 area and DG of LR restraint animals compared to the HR restraint and LR control rats, respectively. The present results indicate that chronic restraint stress in high anxiety rats attenuated CRF expression in the pPVN and DG, which was probably due to detrimental actions on the hippocampus-hypothalamus-pituitary-adrenal gland feedback mechanism, thus modulating the stress response and inducing anhedonia and depressive-like symptoms. PMID:27150225

  2. A novel chiral aliphatic-aromatic diamine promoted direct, highly enantio- and diastereoselective Michael addition of cyclohexanone to nitroolefins under solvent-free conditions.

    PubMed

    Miao, Shifeng; Bai, Jinjin; Yang, Jin; Zhang, Yawen

    2010-10-01

    A series of new highly efficient chiral aliphatic-aromatic diamine catalysts have been designed and successfully applied to the asymmetric Michael addition of cyclohexanone with nitroolefins under solvent-free conditions without any acidic additives. The desired adducts were obtained in high yields with excellent enantio- and diastereoselectivities of syn products (up to >99% ee, >99:1 dr).

  3. The Use of Expressive Arts as Experienced by High School Counselor Trainees

    ERIC Educational Resources Information Center

    Wilson, Barb; Ziomek-Daigle, Jolie

    2013-01-01

    This article presents findings of a qualitative study that explored the process of using expressive arts counseling techniques as experienced by high school counselor trainees. The researcher identified themes that were developed from using grounded theory methods. The three-stage theory, as experienced by high school counseling trainees, included…

  4. High-throughput Cloning and Expression of Integral Membrane Proteins in Escherichia coli

    PubMed Central

    Bruni, Renato

    2014-01-01

    Recently, several structural genomics centers have been established and a remarkable number of three-dimensional structures of soluble proteins have been solved. For membrane proteins, the number of structures solved has been significantly trailing those for their soluble counterparts, not least because over-expression and purification of membrane proteins is a much more arduous process. By using high throughput technologies, a large number of membrane protein targets can be screened simultaneously and a greater number of expression and purification conditions can be employed, leading to a higher probability of successfully determining the structure of membrane proteins. This unit describes the cloning, expression and screening of membrane proteins using high throughput methodologies developed in our laboratory. Basic Protocol 1 deals with the cloning of inserts into expression vectors by ligation-independent cloning. Basic Protocol 2 describes the expression and purification of the target proteins on a miniscale. Lastly, for the targets that express at the miniscale, basic protocols 3 and 4 outline the methods employed for the expression and purification of targets at the midi-scale, as well as a procedure for detergent screening and identification of detergent(s) in which the target protein is stable. PMID:24510647

  5. Endothelial lipase is highly expressed in macrophages in advanced human atherosclerotic lesions.

    PubMed

    Bartels, Emil D; Nielsen, John E; Lindegaard, Marie L S; Hulten, Lillemor M; Schroeder, Torben V; Nielsen, Lars B

    2007-12-01

    Endothelial lipase (EL) is expressed in endothelial cells, and affects plasma lipoprotein metabolism by hydrolyzing phospholipids in HDL. To determine the cellular expression of EL mRNA and protein in human atherosclerotic lesions, we performed in situ hybridization and immunohistochemical studies on sections of carotid endarterectomy specimens from patients with symptomatic cerebrovascular disease. In each of eight patients, EL mRNA and/or protein were seen in areas between the necrotic core and the fibrotic cap where they colocalized with LPL and macrophage-specific CD68. Moreover, there was a positive association between the expression of EL mRNA and CD68 mRNA in plaques from 26 patients. The impact of differentiation from monocytes into macrophages, and subsequently foam cells (by incubation with acetylated LDL) on expression was studied using THP-1 monocytes and primary human monocytes. EL mRNA expression increased markedly when either type of monocytes was differentiated into macrophages. Upon further differentiation into foam cells EL mRNA decreased whereas protein levels remained high compared to monocytes. In conclusion, macrophages in advanced human atherosclerotic lesions display high levels of EL expression, and the level of EL expression varies greatly during transformation of blood monocytes into foam cells.

  6. Ensuring critical event sequences in high consequence computer based systems as inspired by path expressions

    SciTech Connect

    Kidd, M.E.C.

    1997-02-01

    The goal of our work is to provide a high level of confidence that critical software driven event sequences are maintained in the face of hardware failures, malevolent attacks and harsh or unstable operating environments. This will be accomplished by providing dynamic fault management measures directly to the software developer and to their varied development environments. The methodology employed here is inspired by previous work in path expressions. This paper discusses the perceived problems, a brief overview of path expressions, the proposed methods, and a discussion of the differences between the proposed methods and traditional path expression usage and implementation.

  7. High Throughput Gene Expression Measurement with Real Time PCR in a Microfluidic Dynamic Array

    PubMed Central

    Spurgeon, Sandra L.; Jones, Robert C.; Ramakrishnan, Ramesh

    2008-01-01

    We describe a high throughput gene expression platform based on microfluidic dynamic arrays. This system allows 2,304 simultaneous real time PCR gene expression measurements in a single chip, while requiring less pipetting than is required to set up a 96 well plate. We show that one can measure the expression of 45 different genes in 18 tissues with replicates in a single chip. The data have excellent concordance with conventional real time PCR and the microfluidic dynamic arrays show better reproducibility than commercial DNA microarrays. PMID:18301740

  8. Cocaine-induced c-Fos expression in rats selectively bred for high or low saccharin intake and in rats selected for high or low impulsivity.

    PubMed

    Regier, Paul S; Carroll, Marilyn E; Meisel, Robert L

    2012-08-01

    Sweet preference and impulsivity are predictors of cocaine self-administration; however, no research has been conducted to investigate neuronal activation in key brain reward areas after first time exposure to cocaine in rats that differ in their propensity for cocaine-seeking and -taking behavior. In this study we used rats that had been selectively bred for high vs. low saccharin intake and rats selected for high vs. low impulsivity for food. The goal of this study was to investigate whether there are differences of c-Fos reactivity between high and low phenotypes and determine whether these differences are similar between the two animal models. A group of rats was bred for high or low saccharin intake. Another group of rats was selected as high or low impulsive based on performance in a delay-discounting task. Subsequently, rats were given an acute injection of cocaine or saline and then c-Fos expression was observed and analyzed in several brain regions. The low reward-seeking phenotypes showed higher cocaine-induced c-Fos expression in several of these regions. Low saccharin preferring rats showed higher cocaine-induced c-Fos expression in the nucleus accumbens shell, and low impulsive rats showed higher cocaine-induced c-Fos expression in the orbitofrontal cortex and cingulate gyrus 1 area. In addition, both low impulsive and low saccharin rats had higher cocaine-induced c-Fos in the dorsal medial and dorsal lateral caudate putamen. The results indicate that individual differences in neuronal reactivity exist prior to chronic exposure to drugs of abuse. Furthermore, similar differences between the two animal models may be indicative of a common mechanism underlying vulnerability to drugs of abuse.

  9. Food additives

    PubMed Central

    Spencer, Michael

    1974-01-01

    Food additives are discussed from the food technology point of view. The reasons for their use are summarized: (1) to protect food from chemical and microbiological attack; (2) to even out seasonal supplies; (3) to improve their eating quality; (4) to improve their nutritional value. The various types of food additives are considered, e.g. colours, flavours, emulsifiers, bread and flour additives, preservatives, and nutritional additives. The paper concludes with consideration of those circumstances in which the use of additives is (a) justified and (b) unjustified. PMID:4467857

  10. Predicted highly expressed genes in Nocardia farcinica and the implication to its primary metabolism and nocardial virulence

    SciTech Connect

    Wu, Gang; Nie, Lei; Zhang, Weiwen

    2006-02-23

    Nocardia farcinica is a gram positive, filamentous bacterium, and is considered an opportunistic pathogen. In this study, the highly expressed genes in N. farcinica were predicted using the codon adaptation index (CAI) as a numerical estimator of gene expressivity. Using ribosomal protein (RP) genes as references, the top {approx}10% of the genes were predicted to be the predicted highly expressed (PHX) genes in N. farcinica using a CAI cutoff of greater than 0.73. Consistent with early analysis in Streptomyces genomes, most of the PHX genes in N. farcinica were involved in various ''house-keeping'' functions important for cell growth. However, fifteen genes putatively involved in no cardial virulence were predicted as PHX in N. farcinica, which included genes encoding four Mce virulence proteins, cyclopropane fatty acid synthase which is involved in the modification of cell wall important for nocardia virulence, polyketide synthase PKS13 for mycolic acid synthesis and non-ribosomal peptide synthetase involved in biosynthesis of a mycobactin-related siderophore. In addition, multiple genes involved in defense against reactive oxygen species (ROS) produced by the phagocyte were predicted with high expressivity, which included alkylhydroperoxide reductase (ahpC), catalase (katG), superoxide dismutase (sodF), thioredoxin, thioredoxin reductase, glutathione peroxidase, and peptide methionine sulfoxide reductase, suggesting that combating against ROS was essential for survival of N. farcinica in host cells. The study also showed that the distribution of PHX genes in the N. farcinica circular chromosome was uneven, with more PHX genes located in the regions close to replication initiation site. The results provided the first approximates of global gene expression patterns in N. farcinica, which will be useful in guiding experimental design for further investigation.

  11. Effects of dust additions on phytoplankton growth and DMS production in high CO2 northeast Pacific HNLC waters

    NASA Astrophysics Data System (ADS)

    Mélançon, J.; Levasseur, M.; Lizotte, M.; Scarratt, M.; Tremblay, J.-É.; Tortell, P.; Yang, G.-P.; Shi, G.-Y.; Gao, H.-W.; Semeniuk, D. M.; Robert, M.; Arychuk, M.; Johnson, K.; Sutherland, N.; Davelaar, M.; Nemcek, N.; Peña, A.; Richardson, W.

    2015-08-01

    Ocean acidification (OA) is likely to have an effect on the fertilizing potential of desert dust in high-nutrient, low-chlorophyll oceanic regions, either by modifying Fe speciation and bioavailability, or by altering phytoplankton Fe requirements and acquisition. To address this issue, short incubations (4 days) of northeast subarctic Pacific waters enriched with either FeSO4 or dust, and set at pH 8.0 (in situ) and 7.8 were conducted in August 2010. We assessed the impact of a decrease in pH on dissolved Fe concentration, phytoplankton biomass, taxonomy and productivity, and the production of dimethylsulfide (DMS) and its algal precursor dimethylsulfoniopropionate (DMSP). Chlorophyll a (chl a) remained unchanged in the controls and doubled in both the FeSO4-enriched and dust-enriched incubations, confirming the Fe-limited status of the plankton assemblage during the experiment. In the acidified treatments, a significant reduction (by 16-38 %) of the final concentration of chl a was measured compared to their non-acidified counterparts, and a 15 % reduction in particulate organic carbon (POC) concentration was measured in the dust-enriched acidified treatment compared to the dust-enriched non-acidified treatment. FeSO4 and dust additions had a fertilizing effect mainly on diatoms and cyanobacteria. Lowering the pH affected mostly the haptophytes, but pelagophyte concentrations were also reduced in some acidified treatments. Acidification did not significantly alter DMSP and DMS concentrations. These results show that dust deposition events in a low-pH iron-limited Northeast subarctic Pacific are likely to stimulate phytoplankton growth to a lesser extent than in today's ocean during the few days following fertilization and point to a low initial sensitivity of the DMSP and DMS dynamics to OA.

  12. Additional Reserve Recovery Using New Polymer Treatment on High Water Oil Ratio Wells in Alameda Field, Kingman County, Kansas

    SciTech Connect

    James Spillane

    2005-10-01

    The Chemical Flooding process, like a polymer treatment, as a tertiary (enhanced) oil recovery process can be a very good solution based on the condition of this field and its low cost compared to the drilling of new wells. It is an improved water flooding method in which high molecular-weight (macro-size molecules) and water-soluble polymers are added to the injection water to improve the mobility ratio by enhancing the viscosity of the water and by reducing permeability in invaded zones during the process. In other words, it can improve the sweep efficiency by reducing the water mobility. This polymer treatment can be performed on the same active oil producer well rather than on an injector well in the existence of strong water drive in the formation. Some parameters must be considered before any polymer job is performed such as: formation temperature, permeability, oil gravity and viscosity, location and formation thickness of the well, amount of remaining recoverable oil, fluid levels, well productivity, water oil ratio (WOR) and existence of water drive. This improved oil recovery technique has been used widely and has significant potential to extend reservoir life by increasing the oil production and decreasing the water cut. This new technology has the greatest potential in reservoirs that are moderately heterogeneous, contain moderately viscous oils, and have adverse water-oil mobility ratios. For example, many wells in Kansas's Arbuckle formation had similar treatments and we have seen very effective results. In addition, there were previous polymer treatments conducted by Texaco in Alameda Field on a number of wells throughout the Viola-Simpson formation in the early 70's. Most of the treatments proved to be very successful.

  13. Alkyne-azide cycloadditions with copper powder in a high-pressure continuous-flow reactor: high-temperature conditions versus the role of additives.

    PubMed

    Ötvös, Sándor B; Mándity, István M; Kiss, Lóránd; Fülöp, Ferenc

    2013-04-01

    A safe and efficient flow-chemistry-based procedure is presented for 1,3-dipolar cycloaddition reactions between organic azides and acetylenes. This simple and inexpensive technique eliminates the need for costly special apparatus and utilizes Cu powder as a plausible Cu(I) source. To maximize the reaction rates, high-pressure/high-temperature conditions are utilized; alternatively, the harsh reaction conditions can be moderated at room temperature by the joint application of basic and acidic additives. A comparison of the performance of these two approaches in a series of model reactions has resulted in the formation of useful 1,4-disubstituted 1,2,3-triazoles in excellent yields. The risks that are associated with the handling of azides are lowered, thanks to the benefits of flow processing, and gram-scale production has been safely implemented. The synthetic capability of this continuous-flow technique is demonstrated by the efficient syntheses of some highly functionalized derivatives of the antifungal cispentacin.

  14. BacMam System for High-Level Expression of Recombinant Soluble and Membrane Glycoproteins for Structural Studies

    PubMed Central

    Dukkipati, Abhiram; Park, Hyun Ho; Waghray, Deepa; Fischer, Suzanne; Garcia, K. Christopher

    2008-01-01

    Baculovirus mediated gene transduction of mammalian cells (BacMam) is an emerging technique for rapid recombinant protein expression in mammalian cells. Towards this, we constructed two baculovirus transfer vectors that incorporate several mammalian transcriptional regulatory elements necessary for high level protein expression in mammalian cells. Using these vectors, we show that the BacMam system in combination with the 293 GnTI− cell line can be used for production of milligram quantities of soluble glycoproteins. Moreover, for crystallization trials, the purified glycoproteins are sensitive to EndoH treatment resulting in a loss of the bulk of the attached N-linked glycosylation. In addition, we also show that a combination of the BacMam system and 293GnTI− cell line can be used for producing milligram quantities of a GPCR-protein ligand complex suitable for crystallization trials. PMID:18782620

  15. Jasmonates induce nonapoptotic death in high-resistance mutant p53-expressing B-lymphoma cells.

    PubMed

    Fingrut, Orit; Reischer, Dorit; Rotem, Ronit; Goldin, Natalia; Altboum, Irit; Zan-Bar, Israel; Flescher, Eliezer

    2005-11-01

    Mutations in p53, a tumor suppressor gene, occur in more than half of human cancers. Therefore, we tested the hypothesis that jasmonates (novel anticancer agents) can induce death in mutated p53-expressing cells. Two clones of B-lymphoma cells were studied, one expressing wild-type (wt) p53 and the other expressing mutated p53. Jasmonic acid and methyl jasmonate (0.25-3 mM) were each equally cytotoxic to both clones, whereas mutant p53-expressing cells were resistant to treatment with the radiomimetic agent neocarzinostatin and the chemotherapeutic agent bleomycin. Neocarzinostatin and bleomycin induced an elevation in the p53 levels in wt p53-expressing cells, whereas methyl jasmonate did not. Methyl jasmonate induced mostly apoptotic death in the wt p53-expressing cells, while no signs of early apoptosis were detected in mutant p53-expressing cells. In contrast, neocarzinostatin and bleomycin induced death only in wt p53-expressing cells, in an apoptotic mode. Methyl jasmonate induced a rapid depletion of ATP in both clones. In both clones, oligomycin (a mitochondrial ATP synthase inhibitor) did not increase ATP depletion induced by methyl jasmonate, whereas inhibition of glycolysis with 2-deoxyglucose did. High glucose levels protected both clones from methyl jasmonate-induced ATP depletion (and reduced methyl jasmonate-induced cytotoxicity), whereas high levels of pyruvate did not. These results suggest that methyl jasmonate induces ATP depletion mostly by compromising oxidative phosphorylation in the mitochondria. In conclusion, jasmonates can circumvent the resistance of mutant p53-expressing cells towards chemotherapy by inducing a nonapoptotic cell death.

  16. Effects of high fat diet on GPR109A and GPR81 gene expression.

    PubMed

    Wanders, Desiree; Graff, Emily C; Judd, Robert L

    2012-08-24

    GPR109A (PUMA-G, NIACR1, HCA(2)) and GPR81 (HCA(1)) are G protein-coupled receptors located predominantly on adipocytes that mediate anti-lipolytic effects. These cell surface receptors give the adipocyte the ability to "sense" metabolic changes in the environment and respond through lipolytic regulation and release of products including free fatty acids and pro- or anti-inflammatory adipokines. The endogenous ligands for GPR109A and GPR81 are β-hydroxybutyrate and lactate, respectively, both of which are hydroxycarboxylic acids and intermediates of energy metabolism. Circulating β-hydroxybutyrate levels are increased during a 2-3 day fast and prolonged starvation, while lactate levels are elevated during times of intense exercise. Therefore, regulation of expression of these receptors is crucial for the metabolic sensing ability of the adipocyte and ultimately whole body energy homeostasis. We investigated the effects of high fat diet-induced obesity on expression of GPR109A and GPR81. Sixteen male C57BL/6 mice were placed on a control (10% kcal fat; n=8) or a high fat (60% kcal fat; n=8) diet for 11 weeks. Diet-induced obesity significantly reduced GPR109A and GPR81 gene expression in epididymal fat pads. This decrease in GPR109A and GPR81 gene expression was positively correlated with a decrease in adipose tissue PPARγ gene expression. In contrast, acute treatment of both 3T3-L1 adipocytes and RAW 264.7 macrophages with lipopolysaccharide significantly increased GPR109A gene expression, but had no effect on GPR81 expression in 3T3-L1 adipocytes. In conclusion, chronic obesity reduces GPR109A and GPR81 expression in the adipose tissue, while acute in vitro LPS treatment increases expression of GPR109A in adipocytes and macrophages.

  17. Government regulation and public opposition create high additional costs for field trials with GM crops in Switzerland.

    PubMed

    Bernauer, Thomas; Tribaldos, Theresa; Luginbühl, Carolin; Winzeler, Michael

    2011-12-01

    Field trials with GM crops are not only plant science experiments. They are also social experiments concerning the implications of government imposed regulatory constraints and public opposition for scientific activity. We assess these implications by estimating additional costs due to government regulation and public opposition in a recent set of field trials in Switzerland. We find that for every Euro spent on research, an additional 78 cents were spent on security, an additional 31 cents on biosafety, and an additional 17 cents on government regulatory supervision. Hence the total additional spending due to government regulation and public opposition was around 1.26 Euros for every Euro spent on the research per se. These estimates are conservative; they do not include additional costs that are hard to monetize (e.g. stakeholder information and dialogue activities, involvement of various government agencies). We conclude that further field experiments with GM crops in Switzerland are unlikely unless protected sites are set up to reduce these additional costs.

  18. IL-18 Expression Results in a Recombinant Vaccinia Virus That Is Highly Attenuated and Immunogenic

    PubMed Central

    Verardi, Paulo H.; Legrand, Fatema A.; Chan, Kenneth S.; Peng, Yue; Jones, Leslie A.

    2014-01-01

    Interferon-γ (IFN-γ) is an attenuating factor for vaccinia virus (VACV), decreasing its virulence in vivo by more than a million fold. It is also a highly effective adjuvant when administered at the time of immunization with protein antigens. However, recombinant VACV (rVACV) vaccines expressing IFN-γ do not induce enhanced immune responses. It is possible that the IFN-γ expressed by rVACVs induces both an antiviral state and increased immunological clearance, thus resulting in decreased levels of antigen expression due to reduced viral replication and spread. We conjectured that delaying expression of IFN-γ would result in enhanced production of antigens by rVACVs thus resulting in increased immune responses to foreign antigens. Interleukin (IL)-18, also known as IFN-γ inducing factor, is a cytokine that induces T and NK cells to produce IFN-γ. In this study, we demonstrated that an rVACV expressing bioactive murine IL-18 replicated to low but detectable levels in vivo, unlike an rVACV expressing IFN-γ. Moreover, the rVACV expressing IL-18 was significantly attenuated in both immunocompromised and immunocompetent mice. This attenuation was dependent on IFN-γ, as IL-18 expression failed to attenuate VACV in IFN-γ knock-out mice. Cytotoxic T-cell (CTL) and anamnestic antibody responses were slightly increased in animals vaccinated with the rVACV expressing IL-18. Thus, induction of IFN-γ because of IL-18 expression resulted in an rVACV that replicated to low but detectable levels in vivo, yet elicited slightly better CTL and anamnestic humoral immune responses. PMID:24168450

  19. Impact of a non-meltable additive on melt agglomeration with a hydrophobic meltable binder in high-shear mixer.

    PubMed

    Cheong, Wai See; Heng, Paul Wan Sia; Wong, Tin Wui

    2007-01-01

    The present study aims to investigate the behavior of melt agglomeration with a low-viscosity hydrophobic meltable binder by using a non-meltable additive. The size, crushing strength, and pore size distribution of resultant agglomerates, the rheological, surface tension, and wetting properties of the molten binder, as well as, the flow characteristics of preagglomeration powder blend were determined. The use of additive showed contradictory agglomerate growth-promoting and -retarding effects on the molten binder surface tension and the interparticulate frictional forces. Critical concentration effects of additive corresponded to threshold transition of agglomeration-promoting to -retarding behavior were discussed. PMID:17763142

  20. Orotate phosphoribosyltransferase is overexpressed in malignant pleural mesothelioma: Dramatically responds one case in high OPRT expression

    PubMed Central

    Hamamoto, Yoichiro; Takeoka, Shinjiro; Mouri, Atsuto; Fukusumi, Munehisa; Wakuda, Kazushige; Ibe, Tatsuya; Honma, Chie; Arimoto, Yoshihito; Yamada, Kazuaki; Wagatsuma, Miyuki; Tashiro, Akito; Kamoshida, Shingo; Kamimura, Mitsuhiro

    2016-01-01

    ABSTRACT Objective: Malignant pleural mesothelioma (MPM) is a rare and aggressive, treatment-resistant cancer. Pemetrexed, an inhibitor of thymidylate synthase (TS), is used worldwide for MPM as a first-line chemotherapy regimen. However, there is little consensus for a second-line chemotherapy. S-1, a highly effective dihydropyrimidine dehydrogenase (DPD)-inhibitory fluoropyrimidine, mainly acts via a TS inhibitory mechanism similar to pemetrexed. Orotate phosphoribosyltransferase (OPRT) is a key enzyme related to the first step activation of 5-fluorouracil (5-FU) for inhibiting RNA synthesis. We investigated 5-FU related-metabolism proteins, especially focusing on OPRT expression, in MPM Methods and Patients: Fifteen MPM patients who were diagnosed between July 2004 and December 2013 were enrolled. We examined the protein levels of 5-FU metabolism-related enzymes (TS, DPD, OPRT, and thymidine phosphorylase [TP]) in 14 cases Results: High TS, DPD, OPRT, and TP expressions were seen in 28.6%, 71.4%, 85.7%, and 35.7% of patients, respectively. We found that OPRT expression was extremely high in MPM tissue. We experienced one remarkable case of highly effective S-1 combined therapy for pemetrexed refractory MPM. This case also showed high OPRT protein expression Conclusion: The present study suggests that OPRT expression is high in MPM tumors. Although pemetrexed is mainly used for MPM chemotherapy as a TS inhibitor, S-1 has potential as an anticancer drug not only as a TS inhibitor but also inhibiting RNA synthesis through the OPRT pathway. This is the first report investigating OPRT protein expressions in MPM. PMID:27274438

  1. 30 CFR 250.807 - Additional requirements for subsurface safety valves and related equipment installed in high...

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... valves and related equipment installed in high pressure high temperature (HPHT) environments. 250.807... installed in high pressure high temperature (HPHT) environments. (a) If you plan to install SSSVs and related equipment in an HPHT environment, you must submit detailed information with your Application...

  2. 30 CFR 250.807 - Additional requirements for subsurface safety valves and related equipment installed in high...

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... valves and related equipment installed in high pressure high temperature (HPHT) environments. 250.807... installed in high pressure high temperature (HPHT) environments. (a) If you plan to install SSSVs and related equipment in an HPHT environment, you must submit detailed information with your Application...

  3. 30 CFR 250.807 - Additional requirements for subsurface safety valves and related equipment installed in high...

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... valves and related equipment installed in high pressure high temperature (HPHT) environments. 250.807... installed in high pressure high temperature (HPHT) environments. (a) If you plan to install SSSVs and related equipment in an HPHT environment, you must submit detailed information with your Application...

  4. 30 CFR 250.807 - Additional requirements for subsurface safety valves and related equipment installed in high...

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... valves and related equipment installed in high pressure high temperature (HPHT) environments. 250.807... related equipment installed in high pressure high temperature (HPHT) environments. (a) If you plan to install SSSVs and related equipment in an HPHT environment, you must submit detailed information with...

  5. TGFBI protein high expression predicts poor prognosis in colorectal cancer patients

    PubMed Central

    Zhu, Jing; Chen, Xijun; Liao, Zhongcai; He, Chao; Hu, Xiaotong

    2015-01-01

    Transforming growth factor-beta-induced (TGFBI) serves as a linker protein and plays a role in the activation of morphogenesis, cell proliferation, adhesion, migration, differentiation and inflammation. High expression levels of the human TGFBI gene are correlated with numerous human malignancies. In order to explore the roles of TGFBI in the tumor progression of colorectal cancer, colorectal cancer specimens from 115 patients with strict follow-up were selected for the analysis of TGFBI by immunohistochemistry. The correlations between TGFBI expression and the clinicopathological features of colorectal cancers were evaluated. In the colorectal cancer tissues, TGFBI was mainly localized in the cytoplasm and stroma and scarcely in the nucleus. TGFBI expression in the cytoplasm and stroma was not found to be associated with age, gender, tumor histopathological grading, PT category and tumor location (P > 0.05 for each). However, high TGFBI expression in the cytoplasm and stroma correlated with lymph node metastasis, distant metastasis and Dukes stage (P < 0.05 for each). The survival rate was significantly lower in patients with high TGFBI expression than in those with low TGFBI expression. Furthermore, we found that tumor node metastasis (TNM) staging (HR: 2.963; 95% CI: 1.573-1.664; P = 0.000), differentiation (HR: 1.574; 95% CI: 1.001-2.476; P = 0.049) and high TGFBI cytoplasmic expression (HR: 3.332; 95% CI: 1.410-7.873; P = 0.000) proved to be independent prognostic factors for survival in colorectal cancer. In conclusion, TGFBI plays an important role in the progression of colorectal cancers and it is an independent poor prognostic factor for colorectal cancer patients. PMID:25755764

  6. HIGHLY DIASTEREOSELECTIVE MICHAEL REACTION UNDER SOLVENT-FREE CONDITIONS USING MICROWAVES: CONJUGATE ADDITION OF FLAVANONE TO ITS CHALCONE PRECURSOR

    EPA Science Inventory

    Microwave-assisted reaction of 2'-hydroxychalcones in the presence of DBU resulted in the formation of hitherto unknown dimers by conjugate addition of the intermediate cyclic ketone to the starting enone.

  7. Development of High-Antifouling PPSU Ultrafiltration Membrane by Using Compound Additives: Preparation, Morphologies, and Filtration Resistant Properties.

    PubMed

    Liu, Jie; Zhong, Zhencheng; Ma, Rui; Zhang, Weichen; Li, Jiding

    2016-01-01

    In this study, flat sheet asymmetric polyphenylsulfone (PPSU) ultrafiltration membranes with enhanced antifouling properties were prepared with a non-solvent induced phase separation (NIPS) method through compound additives containing a polymeric pore-forming agent, a small molecular non-solvent and a surfactant. The formation processes of the porous asymmetric membranes with different kinds of additives were studied in detail, and the microstructure controllable preparation of membrane was achieved by establishing a bridge between the membrane preparation parameters and separation performances. All prepared membranes were characterized by using a scanning electron microscope (SEM), contact angle analysis, porosity, maximum pore size, water and BSA solution permeability studies. The performance efficiency of the membrane was evaluated by using BSA as a model foulant in terms of permeability, solute rejection (R), Rm (membrane inherent resistance), Rc (cake layer resistance), and Rp (pore plugging resistance). The results showed that when the compound additives were used, the inter-connected pores were observed, maximum pore size, contact angle and membrane filtration resistance decreased, while the porosity increased. When PVP compound additives were added, the water flux increased from 80.4 to 148.1 L/(m²·h), the BSA rejection increased from 53.2% to 81.5%. A similar trend was observed for membranes with added PEG compound additives; the water flux and BSA rejection simultaneously increased. The filtration resistance decreased as a result of compound additives. The uniformity of membrane and the number of effective pores could be enhanced by adding compound additives through the cooperation of different additives.

  8. Development of High-Antifouling PPSU Ultrafiltration Membrane by Using Compound Additives: Preparation, Morphologies, and Filtration Resistant Properties.

    PubMed

    Liu, Jie; Zhong, Zhencheng; Ma, Rui; Zhang, Weichen; Li, Jiding

    2016-01-01

    In this study, flat sheet asymmetric polyphenylsulfone (PPSU) ultrafiltration membranes with enhanced antifouling properties were prepared with a non-solvent induced phase separation (NIPS) method through compound additives containing a polymeric pore-forming agent, a small molecular non-solvent and a surfactant. The formation processes of the porous asymmetric membranes with different kinds of additives were studied in detail, and the microstructure controllable preparation of membrane was achieved by establishing a bridge between the membrane preparation parameters and separation performances. All prepared membranes were characterized by using a scanning electron microscope (SEM), contact angle analysis, porosity, maximum pore size, water and BSA solution permeability studies. The performance efficiency of the membrane was evaluated by using BSA as a model foulant in terms of permeability, solute rejection (R), Rm (membrane inherent resistance), Rc (cake layer resistance), and Rp (pore plugging resistance). The results showed that when the compound additives were used, the inter-connected pores were observed, maximum pore size, contact angle and membrane filtration resistance decreased, while the porosity increased. When PVP compound additives were added, the water flux increased from 80.4 to 148.1 L/(m²·h), the BSA rejection increased from 53.2% to 81.5%. A similar trend was observed for membranes with added PEG compound additives; the water flux and BSA rejection simultaneously increased. The filtration resistance decreased as a result of compound additives. The uniformity of membrane and the number of effective pores could be enhanced by adding compound additives through the cooperation of different additives. PMID:27338487

  9. Development of High-Antifouling PPSU Ultrafiltration Membrane by Using Compound Additives: Preparation, Morphologies, and Filtration Resistant Properties

    PubMed Central

    Liu, Jie; Zhong, Zhencheng; Ma, Rui; Zhang, Weichen; Li, Jiding

    2016-01-01

    In this study, flat sheet asymmetric polyphenylsulfone (PPSU) ultrafiltration membranes with enhanced antifouling properties were prepared with a non-solvent induced phase separation (NIPS) method through compound additives containing a polymeric pore-forming agent, a small molecular non-solvent and a surfactant. The formation processes of the porous asymmetric membranes with different kinds of additives were studied in detail, and the microstructure controllable preparation of membrane was achieved by establishing a bridge between the membrane preparation parameters and separation performances. All prepared membranes were characterized by using a scanning electron microscope (SEM), contact angle analysis, porosity, maximum pore size, water and BSA solution permeability studies. The performance efficiency of the membrane was evaluated by using BSA as a model foulant in terms of permeability, solute rejection (R), Rm (membrane inherent resistance), Rc (cake layer resistance), and Rp (pore plugging resistance). The results showed that when the compound additives were used, the inter-connected pores were observed, maximum pore size, contact angle and membrane filtration resistance decreased, while the porosity increased. When PVP compound additives were added, the water flux increased from 80.4 to 148.1 L/(m2·h), the BSA rejection increased from 53.2% to 81.5%. A similar trend was observed for membranes with added PEG compound additives; the water flux and BSA rejection simultaneously increased. The filtration resistance decreased as a result of compound additives. The uniformity of membrane and the number of effective pores could be enhanced by adding compound additives through the cooperation of different additives. PMID:27338487

  10. Genomic Imprinting of IGF2 Is Maintained in Infantile Hemangioma despite its High Level of Expression

    PubMed Central

    Yu, Ying; Wylie-Sears, Jill; Boscolo, Elisa; Mulliken, John B; Bischoff, Joyce

    2004-01-01

    Hemangioma, the most common tumor of infancy, is characterized by rapid growth and slow regression. Increased mRNA expression of insulin-like growth factor 2 (IGF2) has been detected in the proliferating phase by cDNA microarray analysis, but the underlying mechanism causing the increase remains unknown. Here, using quantitative real-time polymerase chain reaction (PCR) and immunohistochemistry, we show that IGF2 is highly expressed in both proliferating and involuting phase hemangioma, but is not detectable in other vascular lesions such as pyogenic granuloma, venous malformation, lymphatic malformation, or in normal infant skin. Loss of imprinting of the Igf2 gene has been associated with IGF2 overexpression in a variety of childhood tumors. To determine if loss of imprinting and consequent bi-allelic expression might contribute to the increased expression of IGF2, we examined the genomic imprinting status of Igf2 in 48 individual hemangiomas. We determined allele-specific Igf2 expression using reverse transcriptase–PCR combined with analysis of an Apa I–sensitive restriction fragment length polymorphism. Similar to heterozygous normal skin controls, all 15 informative hemangiomas showed uniform mono-allelic expression of Igf2. Therefore, loss of imprinting is not involved in the increased expression of IGF2 in infantile hemangioma. PMID:15706404

  11. Development of a new noncytopathic Semliki Forest virus vector providing high expression levels and stability

    SciTech Connect

    Casales, Erkuden; Rodriguez-Madoz, Juan R.; Ruiz-Guillen, Marta; Razquin, Nerea; Cuevas, Yolanda; Prieto, Jesus; Smerdou, Cristian

    2008-06-20

    Alphavirus vectors express high levels of recombinant proteins in mammalian cells, but their cytopathic nature makes this expression transient. In order to generate a Semliki Forest virus (SFV) noncytopathic vector we introduced mutations previously described to turn Sindbis virus noncytopathic into a conserved position in an SFV vector expressing LacZ. Interestingly, mutant P718T in replicase nsp2 subunit was able to replicate in only a small percentage of BHK cells, producing {beta}-gal-expressing colonies without selection. Puromycin N-acetyl-transferase (pac) gene was used to replace LacZ in this mutant allowing selection of an SFV noncytopathic replicon containing a second mutation in nsp2 nuclear localization signal (R649H). This latter mutation did not confer a noncytopathic phenotype by itself and did not alter nsp2 nuclear translocation. Replicase synthesis was diminished in the SFV double mutant, leading to genomic and subgenomic RNA levels that were 125-fold and 66-fold lower than in wild-type vector, respectively. Interestingly, this mutant expressed {beta}-gal levels similar to parental vector. By coexpressing pac and LacZ from independent subgenomic promoters this vector was able to generate stable cell lines maintaining high expression levels during at least 10 passages, indicating that it could be used as a powerful system for protein production in mammalian cells.

  12. Functional analysis of a RING domain ankyrin repeat protein that is highly expressed during flower senescence.

    PubMed

    Xu, Xinjia; Jiang, Cai-Zhong; Donnelly, Linda; Reid, Michael S

    2007-01-01

    A gene encoding a RING zinc finger ankyrin repeat protein (MjXB3), a putative E3 ubiquitin ligase, is highly expressed in petals of senescing four o'clock (Mirabilis jalapa) flowers, increasing >40,000-fold during the onset of visible senescence. The gene has homologues in many other species, and the Petunia homologue is strongly up-regulated in senescing Petunia corollas. Silencing the expression of this gene in Petunia, using virus-induced gene silencing, resulted in a 2 d extension in flower life. In Mirabilis, a 2 kb promoter region, 5' upstream of the MjXB3 gene, was isolated. The promoter sequence included putative binding sites for many DNA-binding proteins, including the bZIP, Myb, homeodomain-leucine zipper (HD-Zip), MADS-box, and WRKY transcription factors. The construct containing a 1 kb promoter region immediately upstream of the MjXB3 gene drove the strongest expression of the beta-glucuronidase (GUS) reporter gene in a transient expression assay. In Petunia, GUS expression under the control of this heterologous promoter fragment was specific to senescing flowers. The Mirabilis promoter GUS construct was tested in other flower species; while GUS activity in carnation petals was high during senescence, no expression was detected in three monocotyledonous flowers--daylily (Hemerocallis 'Stella d'Oro'), daffodil (Narcissus pseudonarcissus 'King Alfred'), and orchid (Dendrobium 'Emma White'). PMID:18057040

  13. Development of a new noncytopathic Semliki Forest virus vector providing high expression levels and stability.

    PubMed

    Casales, Erkuden; Rodriguez-Madoz, Juan R; Ruiz-Guillen, Marta; Razquin, Nerea; Cuevas, Yolanda; Prieto, Jesus; Smerdou, Cristian

    2008-06-20

    Alphavirus vectors express high levels of recombinant proteins in mammalian cells, but their cytopathic nature makes this expression transient. In order to generate a Semliki Forest virus (SFV) noncytopathic vector we introduced mutations previously described to turn Sindbis virus noncytopathic into a conserved position in an SFV vector expressing LacZ. Interestingly, mutant P718T in replicase nsp2 subunit was able to replicate in only a small percentage of BHK cells, producing beta-gal-expressing colonies without selection. Puromycin N-acetyl-transferase (pac) gene was used to replace LacZ in this mutant allowing selection of an SFV noncytopathic replicon containing a second mutation in nsp2 nuclear localization signal (R649H). This latter mutation did not confer a noncytopathic phenotype by itself and did not alter nsp2 nuclear translocation. Replicase synthesis was diminished in the SFV double mutant, leading to genomic and subgenomic RNA levels that were 125-fold and 66-fold lower than in wild-type vector, respectively. Interestingly, this mutant expressed beta-gal levels similar to parental vector. By coexpressing pac and LacZ from independent subgenomic promoters this vector was able to generate stable cell lines maintaining high expression levels during at least 10 passages, indicating that it could be used as a powerful system for protein production in mammalian cells.

  14. High LIFr expression stimulates melanoma cell migration and is associated with unfavorable prognosis in melanoma.

    PubMed

    Guo, Hongwei; Cheng, Yabin; Martinka, Magdalena; McElwee, Kevin

    2015-09-22

    Increased or decreased expression of LIF receptor (LIFr) has been reported in several human cancers, including skin cancer, but its role in melanoma is unknown. In this study, we investigated the expression pattern of LIFr in melanoma and assessed its prognostic value. Using tissue microarrays consisting of 441 melanomas and 96 nevi, we found that no normal nevi showed high LIFr expression. LIFr staining was significantly increased in primary melanoma compared to dysplastic nevi (P = 0.0003) and further increased in metastatic melanoma (P = 0.0000). Kaplan-Meier survival curve and univariate Cox regression analyses showed that increased expression of LIFr was correlated with poorer 5-year patient survival (overall survival, P = 0.0000; disease-specific survival, P = 0.0000). Multivariate Cox regression analyses indicated that increased LIFr expression was an independent prognostic marker for primary melanoma (P = 0.036). LIFr knockdown inhibited melanoma cell migration in wound healing assays and reduced stress fiber formation. LIFr knockdown correlated with STAT3 suppression, but not YAP, suggesting that LIFr activation might stimulate melanoma cell migration through the STAT3 pathway. Our data indicate that strong LIFr expression identifies potentially highly malignant melanocytic lesions at an early stage and LIFr may be a potential target for the development of early intervention therapeutics. PMID:26329521

  15. Evaluation of two intracavitary high-dose-rate brachytherapy devices for irradiating additional and irregularly shaped volumes of breast tissue

    SciTech Connect

    Lu, Sharon M.; Scanderbeg, Daniel J.; Barna, Patrick; Yashar, William; Yashar, Catheryn

    2012-04-01

    The SAVI and Contura breast brachytherapy applicators represent 2 recent advancements in brachytherapy technology that have expanded the number of women eligible for accelerated partial breast irradiation in the treatment of early-stage breast cancer. Early clinical experience with these 2 single-entry, multichannel high-dose-rate brachytherapy devices confirms their ease of use and dosimetric versatility. However, current clinical guidelines for SAVI and Contura brachytherapy may result in a smaller or less optimal volume of treated tissue compared with traditional interstitial brachytherapy. This study evaluates the feasibility of using the SAVI and Contura to irradiate larger and irregularly shaped target volumes, approaching what is treatable with the interstitial technique. To investigate whether additional tissue can be treated, 17 patients treated with the SAVI and 3 with the Contura were selected. For each patient, the planning target volume (PTV) was modified to extend 1.1 cm, 1.3 cm, and 1.5 cm beyond the tumor bed cavity. To evaluate dose conformance to an irregularly shaped target volume, 9 patients treated with the SAVI and 3 with the Contura were selected from the original 20 patients. The following asymmetric PTV margin combinations were assessed for each patient: 1.5/0.3, 1.3/0.3, and 1.1/0.3 cm. For all patients, treatment planning was performed, adopting the National Surgical Adjuvant Breast and Bowel Project guidelines, and dosimetric comparisons were made. The 6-1 and 8-1 SAVI devices can theoretically treat a maximal tissue margin of 1.5 cm and an asymmetric PTV with margins ranging from 0.3 to 1.5 cm. The 10-1 SAVI and Contura can treat a maximal margin of 1.3 cm and 1.1 cm, respectively, and asymmetric PTV with margins ranging from 0.3-1.3 cm. Compared with the Contura, the SAVI demonstrated greater dosimetric flexibility. Risk of developing excessive hot spots increased with the size of the SAVI device. Both the SAVI and Contura appear

  16. Structure and high-temperature properties of Ti{sub 5}Si{sub 3} with interstitial additions

    SciTech Connect

    Williams, Jason

    1999-12-01

    This study was motivated by the fact that previous research on the structure and properties of Ti{sub 5}Si{sub 3} showed unacceptably inconsistent results. The primary reason for these inconsistencies was interstitial contamination of Ti{sub 5}Si{sub 3} by carbon, nitrogen and oxygen. Thus, this study measured the effects that these interstitial atoms have on some of the previously reported properties. These properties include crystalline structure, thermal expansion anisotropy, electronic structure and bonding, and high temperature oxidation resistance. In Chapter 2 of this study, the lattice parameters and atomic positions of Ti{sub 5}Si{sub 3} as a function of carbon, nitrogen or oxygen content were measured via x-ray and neutron diffraction. Comparing these lattice parameters to those reported in other studies on supposedly pure Ti{sub 5}Si{sub 3} confirmed that the majority of the previous studies had samples with a considerable amount of interstitial impurities. In fact, the lattice parameter trends given in Chapter 2 can be used to estimate the types and level of impurities in these studies. Furthermore, Chapter 2 discusses how atomic positions change as interstitial atoms are incorporated into the lattice. These changes in atomic separations suggest that strong bonds form between the interstitial atoms and the surrounding titanium atoms. This is in full agreement with the electronic structure calculations given in Chapter 4. These calculations show that bonding does occur between titanium d-states and interstitial atom p-states at the expense of bonding between some of the titanium and silicon atoms. In addition, carbon seems to be the most strongly bonded interstitial atom. Knowledge of the exact interstitial content and its effect on bonding is important because Chapters 3 and 5 have shown that interstitial atoms have a marked effect on the thermal expansion and oxidation resistance. As discussed in Chapter 3, all interstitial atoms lower the thermal

  17. Evaluation of two intracavitary high-dose-rate brachytherapy devices for irradiating additional and irregularly shaped volumes of breast tissue.

    PubMed

    Lu, Sharon M; Scanderbeg, Daniel J; Barna, Patrick; Yashar, William; Yashar, Catheryn

    2012-01-01

    The SAVI and Contura breast brachytherapy applicators represent 2 recent advancements in brachytherapy technology that have expanded the number of women eligible for accelerated partial breast irradiation in the treatment of early-stage breast cancer. Early clinical experience with these 2 single-entry, multichannel high-dose-rate brachytherapy devices confirms their ease of use and dosimetric versatility. However, current clinical guidelines for SAVI and Contura brachytherapy may result in a smaller or less optimal volume of treated tissue compared with traditional interstitial brachytherapy. This study evaluates the feasibility of using the SAVI and Contura to irradiate larger and irregularly shaped target volumes, approaching what is treatable with the interstitial technique. To investigate whether additional tissue can be treated, 17 patients treated with the SAVI and 3 with the Contura were selected. For each patient, the planning target volume (PTV) was modified to extend 1.1 cm, 1.3 cm, and 1.5 cm beyond the tumor bed cavity. To evaluate dose conformance to an irregularly shaped target volume, 9 patients treated with the SAVI and 3 with the Contura were selected from the original 20 patients. The following asymmetric PTV margin combinations were assessed for each patient: 1.5/0.3, 1.3/0.3, and 1.1/0.3 cm. For all patients, treatment planning was performed, adopting the National Surgical Adjuvant Breast and Bowel Project guidelines, and dosimetric comparisons were made. The 6-1 and 8-1 SAVI devices can theoretically treat a maximal tissue margin of 1.5 cm and an asymmetric PTV with margins ranging from 0.3 to 1.5 cm. The 10-1 SAVI and Contura can treat a maximal margin of 1.3 cm and 1.1 cm, respectively, and asymmetric PTV with margins ranging from 0.3-1.3 cm. Compared with the Contura, the SAVI demonstrated greater dosimetric flexibility. Risk of developing excessive hot spots increased with the size of the SAVI device. Both the SAVI and Contura appear

  18. DAP1 high expression increases risk of lymph node metastases in squamous cell carcinoma of the oral cavity.

    PubMed

    Santos, M; Maia, L L; Silva, C V M; Peterle, G T; Mercante, A M C; Nunes, F D; Carvalho, M B; Tajara, E H; Louro, I D; Silva-Conforti, A M A

    2015-01-01

    Death-associated protein 1 (DAP1) is a member of the DAP family. Its expression is associated with cell growth and normal death of the neoplastic cells, regulated by the mammalian target of the rapamycin protein. Activated DAP1 negatively regulates autophagy, which has been associated with the development and progression of several diseases, such as cancer, and with prognosis and survival of diverse tumor types. Therefore, in this study we analyzed DAP1 expression in 54 oral squamous cell carcinoma tumor samples and in 20 non-tumoral margins by immunohistochemistry. The results showed that DAP1 is more frequently expressed in tumor tissues compared with marginal non-tumoral cells. Additionally, high DAP1 expression is associated with a 4-fold increase in the risk of lymph node metastases. Our results suggest that the DAP1 protein can be used as a potential marker of lymph node metastases predisposition, helping define the best therapy for each patient to minimize risk of developing metastases. PMID:26400283

  19. Expression of high-affinity IL-4 receptors on human melanoma, ovarian and breast carcinoma cells.

    PubMed Central

    Obiri, N I; Siegel, J P; Varricchio, F; Puri, R K

    1994-01-01

    It has previously been shown that murine sarcoma cells express high-affinity IL-4 receptors (IL-4R) which are internalized after binding to the ligand (Puri et al., Cancer Res 1991; 51:3011-7). We have also reported that human renal cell carcinoma cells express high-affinity IL-4R, and IL-4 inhibits tumour growth in vitro (Obiri et al., J Clin Invest 1993; 91:88). In this study we investigated the expression and function of IL-4R on other human solid tumours. Human melanoma, ovarian carcinoma and breast carcinoma cell lines were assessed for the cell surface expression of IL-4R by radio-ligand receptor binding and for IL-4R gene expression by Northern blot analysis. Primary cultures of mesothelioma and neurofibrosarcoma cells were similarly investigated. Human melanoma, ovarian carcinoma and breast carcinoma cell lines expressed IL-4R on their cell surface with a dissociation constant (Kd) of 140-549 pM. These tumour lines expressed a single 4 kb species of mRNA for IL-4R. Similarly, primary cultures of mesothelioma and neurofibrosarcoma cells were positive for the IL-4R mRNA by Northern blot analysis. Fresh, non-cultured mesothelioma and neurofibrosarcoma tumour sections were also positive for the presence of IL-4R as determined by immunohistochemistry of frozen sections using anti-IL-4R antibody. In order to study possible functions of IL-4R, we evaluated the effects of IL-4 on cell growth and its effect on MHC antigen expression in the presence or absence of interferon-gamma (IFN-gamma). In tissue culture, IL-4 reduced the growth of tumour cell lines and primary cell cultures studied. IL-4 had very little effect on MHC class I antigen expression on ovarian, breast and melanoma cell lines; however, MHC class II (HLA-DR) expression was enhanced on melanoma and breast carcinoma cells. IL-4 also enhanced the IFN-gamma-induced class II expression on melanoma and breast carcinoma cells. Taken together, our observations indicate that IL-4R are expressed on a variety of

  20. Matricellular protein CCN1 (CYR61) expression is associated with high-grade ductal carcinoma in situ.

    PubMed

    Saglam, Ozlen; Dai, Feng; Husain, Seema; Zhan, Yilei; Toruner, Gokce; Haines, G Kenneth

    2014-06-01

    Cysteine-rich protein 61, connective tissue growth factor, and nephroblastoma overexpressed gene (CCN) comprise a family of matricellular proteins that have multiple physiologic functions including development, tissue repair, cell adhesion, migration, and proliferation. The expression of CCN1, cyclin D1, β-catenin, and p53 was explored by immunohistochemistry in different grades of ductal carcinoma in situ (DCIS) cases. These cases did not contain any infiltrating carcinoma components. In addition, all cysteine-rich protein 61 gene exons (encoding the CCN1 protein) were sequenced in 30 samples. Allred and H-scores were calculated for expression in both DCIS and the surrounding benign breast tissue. All cases of DCIS showed degrees of cytoplasmic CCN1 staining with median H-scores of 170, 160, and 60 in grades 3, 2, and 1, respectively (P = .043). Twelve of 28 DCIS 3, 1 of 15 DCIS 2, and 0 of 18 DCIS 1 also showed nuclear staining for CCN1. The cytoplasmic staining difference was preserved when the cases were divided into estrogen receptor (ER)+/DCIS grade 1, ER+/DCIS 2 and 3, and ER-/DCIS 2 and 3 by the H-score (P = .037). Cyclin D1 expression was positively correlated with the CCN1 cytoplasmic H-score in all DCIS samples (P = .038). Membranous β-catenin expression correlated with the grade of intraepithelial carcinoma by both H-score (P = .047) and Allred score (P = .026). Our results suggest that CCN1 has a role in the development of intraepithelial carcinoma. CCN1 expression correlates with grade of DCIS independent of ER status. It can induce cell cycle progression through cyclin D1. It is warranted to study high expression of CCN1 in DCIS as an independent risk factor in a larger cohort.

  1. High expression and biosilica encapsulation of alkaline-active carbonic anhydrase for CO2 sequestration system development.

    PubMed

    Min, Ki-Ha; Son, Ryeo Gang; Ki, Mi-Ran; Choi, Yoo Seong; Pack, Seung Pil

    2016-01-01

    Carbonic anhydrase (CA) is a biocatalyst for CO2 sequestration because of its distinctive ability to accelerate CO2 hydration. High production and efficient immobilization of alkaline-active CAs are required, because one potential application of CA is its use in the alkaline solvent-based CO2 absorption/desorption process. Here, we designed and applied an α-type CA from Hahella chejuensis (HCA), which was reported as highly active in alkaline conditions, but was mostly expressed as insoluble forms. We found that the signal peptide-removed form of HCA [HCA(SP-)] was successfully expressed in the soluble form [∼70mg of purified HCA(SP-) per L of culture]. HCA(SP-) also displayed high pH stability in alkaline conditions, with maximal activity at pH 10; at this pH, ∼90% activity was maintained for 2h. Then, we prepared HCA(SP-)-encapsulated silica particles [HCA(SP-)@silica] via a spermine-mediated bio-inspired silicification method. HCA(SP-)@silica exhibited high-loading and highly stable CA activity. In addition, HCA(SP-)@silica retained more than 90% of the CA activity even after 10 cycles of use in mild conditions, and ∼80% in pH 10 conditions. These results will be useful for the development of practical CO2 sequestration processes employing CA.

  2. High-throughput recombinant protein expression in Escherichia coli: current status and future perspectives.

    PubMed

    Jia, Baolei; Jeon, Che Ok

    2016-08-01

    The ease of genetic manipulation, low cost, rapid growth and number of previous studies have made Escherichia coli one of the most widely used microorganism species for producing recombinant proteins. In this post-genomic era, challenges remain to rapidly express and purify large numbers of proteins for academic and commercial purposes in a high-throughput manner. In this review, we describe several state-of-the-art approaches that are suitable for the cloning, expression and purification, conducted in parallel, of numerous molecules, and we discuss recent progress related to soluble protein expression, mRNA folding, fusion tags, post-translational modification and production of membrane proteins. Moreover, we address the ongoing efforts to overcome various challenges faced in protein expression in E. coli, which could lead to an improvement of the current system from trial and error to a predictable and rational design. PMID:27581654

  3. High-throughput recombinant protein expression in Escherichia coli: current status and future perspectives

    PubMed Central

    2016-01-01

    The ease of genetic manipulation, low cost, rapid growth and number of previous studies have made Escherichia coli one of the most widely used microorganism species for producing recombinant proteins. In this post-genomic era, challenges remain to rapidly express and purify large numbers of proteins for academic and commercial purposes in a high-throughput manner. In this review, we describe several state-of-the-art approaches that are suitable for the cloning, expression and purification, conducted in parallel, of numerous molecules, and we discuss recent progress related to soluble protein expression, mRNA folding, fusion tags, post-translational modification and production of membrane proteins. Moreover, we address the ongoing efforts to overcome various challenges faced in protein expression in E. coli, which could lead to an improvement of the current system from trial and error to a predictable and rational design. PMID:27581654

  4. High fat diet and body weight have different effects on cannabinoid CB1 receptor expression in rat nodose ganglia

    PubMed Central

    Cluny, N.L.; Baraboi, E.D.; Mackie, K; Burdyga, G.; Richard, D.; Dockray, G.J.; Sharkey, K.A.

    2013-01-01

    Energy balance is regulated, in part, by orexigenic signaling pathways of the vagus nerve. Fasting-induced modifications in the expression of orexigenic signaling systems have been observed in vagal afferents of lean animals. Altered basal cannabinoid (CB)1 receptor expression in the nodose ganglia in obesity has been reported. Whether altered body weight or a high fat diet modifies independent or additive changes in CB1 expression is unknown. We investigated the expression of CB1 and orexin 1 receptor (OX-1R) in nodose ganglia of rats fed ad libitum or food deprived (24h), maintained on low or high fat diets (HFD), with differing body weights. Male Wistar rats were fed chow or HFD (diet-induced obese: DIO or diet-resistant: DR) or were body weight matched to the DR group but fed chow (wmDR). CB1 and OX-1R immunoreactivity were investigated and CB1 mRNA density was determined using in situ hybridization. CB1 immunoreactivity was measured in fasted rats after sulfated cholecystokinin octapeptide (CCK8s) administration. In chow rats, fasting did not modify the level of CB1 mRNA. More CB1 immunoreactive cells were measured in fed DIO, DR and wmDR rats than chow rats; levels increased after fasting in chow and wmDR rats but not in DIO or DR rats. In HFD fasted rats CCK8s did not reduce CB1 immunoreactivity. OX-1R immunoreactivity was modified by fasting only in DR rats. These data suggest that body weight contributes to the proportion of neurons expressing CB1 immunoreactivity in the nodose ganglion, while HFD blunts fasting-induced increases, and CCK-induced suppression of, CB1-immunoreactivity. PMID:24145047

  5. High Levels of MeCP2 Depress MHC Class I Expression in Neuronal Cells

    PubMed Central

    Miralvès, Julie; Magdeleine, Eddy; Kaddoum, Lara; Brun, Hélène; Peries, Sophie; Joly, Etienne

    2007-01-01

    Background The expression of MHC class I genes is repressed in mature neurons. The molecular basis of this regulation is poorly understood, but the genes are particularly rich in CpG islands. MeCP2 is a transcriptional repressor that binds to methylated CpG dinucleotides; mutations in this protein also cause the neurodevelopmental disease called Rett syndrome. Because MHC class I molecules play a role in neuronal connectivity, we hypothesised that MeCP2 might repress MHC class I expression in the CNS and that this might play a role in the pathology of Rett syndrome. Methodology We show here that transiently transfected cells expressing high levels of MeCP2 specifically downregulate cell-surface expression of MHC class I molecules in the neuronal cell line N2A and they prevent the induction of MHC class I expression in response to interferon in these cells, supporting our first hypothesis. Surprisingly, however, overexpression of the mutated forms of MeCP2 that cause Rett syndrome had a similar effect on MHC class I expression as the wild-type protein. Immunohistological analyses of brain slices from MECP2 knockout mice (the MeCP2tm1.1Bird strain) demonstrated a small but reproducible increase in MHC class I when compared to their wild type littermates, but we found no difference in MHC class I expression in primary cultures of mixed glial cells (mainly neurons and astrocytes) from the knockout and wild-type mice. Conclusion These data suggest that high levels of MeCP2, such as those found in mature neurons, may contribute to the repression of MHC expression, but we find no evidence that MeCP2 regulation of MHC class I is important for the pathogenesis of Rett syndrome. PMID:18159237

  6. ADAM9 is highly expressed in renal cell cancer and is associated with tumour progression

    PubMed Central

    Fritzsche, Florian R; Wassermann, Kirsten; Jung, Monika; Tölle, Angelika; Kristiansen, Ilka; Lein, Michael; Johannsen, Manfred; Dietel, Manfred; Jung, Klaus; Kristiansen, Glen

    2008-01-01

    Background A Disintegrin And Metalloprotease (ADAM) 9 has been implicated in tumour progression of various solid tumours, however, little is known about its role in renal cell carcinoma. We evaluated the expression of ADAM9 on protein and transcript level in a clinico-pathologically characterized renal cell cancer cohort. Methods 108 renal cancer cases were immunostained for ADAM9 on a tissue-micro-array. For 30 additional cases, ADAM9 mRNA of microdissected tumour and normal tissue was analyzed via quantitative RT-PCR. SPSS 14.0 was used to apply crosstables (Fisher's exact test and χ2-test), correlations and univariate as well as multivariate survival analyses. Results ADAM9 was significantly up-regulated in renal cancer in comparison to the adjacent normal tissue on mRNA level. On protein level, ADAM9 was significantly associated with higher tumour grade, positive nodal status and distant metastasis. Furthermore, ADAM9 protein expression was significantly associated with shortened patient survival in the univariate analysis. Conclusion ADAM9 is strongly expressed in a large proportion of renal cell cancers, concordant with findings in other tumour entities. Additionally, ADAM9 expression is significantly associated with markers of unfavourable prognosis. Whether the demonstrated prognostic value of ADAM9 is independent from other tumour parameters will have to be verified in larger study cohorts. PMID:18582378

  7. Transgenic mice expressing high plasma concentrations of human apolipoprotein B100 and lipoprotein(a).

    PubMed Central

    Linton, M F; Farese, R V; Chiesa, G; Grass, D S; Chin, P; Hammer, R E; Hobbs, H H; Young, S G

    1993-01-01

    The B apolipoproteins, apo-B48 and apo-B100, are key structural proteins in those classes of lipoproteins considered to be atherogenic [e.g., chylomicron remnants, beta-VLDL, LDL, oxidized LDL, and Lp(a)]. Here we describe the development of transgenic mice expressing high levels of human apo-B48 and apo-B100. A 79.5-kb human genomic DNA fragment containing the entire human apo-B gene was isolated from a P1 bacteriophage library and microinjected into fertilized mouse eggs. 16 transgenic founders expressing human apo-B were generated, and the animals with the highest expression had plasma apo-B100 levels nearly as high as those of normolipidemic humans (approximately 50 mg/dl). The human apo-B100 in transgenic mouse plasma was present largely in lipoproteins of the LDL class as shown by agarose gel electrophoresis, chromatography on a Superose 6 column, and density gradient ultracentrifugation. When the human apo-B transgenic founders were crossed with transgenic mice expressing human apo(a), the offspring that expressed both transgenes had high plasma levels of human Lp(a). Both the human apo-B and Lp(a) transgenic mice will be valuable resources for studying apo-B metabolism and the role of apo-B and Lp(a) in atherosclerosis. Images PMID:8254057

  8. Neural activity to positive expressions predicts daily experience of schizophrenia-spectrum symptoms in adults with high social anhedonia.

    PubMed

    Hooker, Christine I; Benson, Taylor L; Gyurak, Anett; Yin, Hong; Tully, Laura M; Lincoln, Sarah Hope

    2014-02-01

    Social anhedonia (SA), the diminished pleasure from social relationships, is a prominent characteristic of the vulnerability and manifestation of schizophrenia disorder. However, SA can develop for multiple reasons and little is known about its neural basis; these 2 issues hinder the utility and sensitivity of SA as a marker of schizophrenia pathology. This study investigated whether lateral prefrontal cortex (LPFC) deficits in social reward processing are associated with both SA and other schizophrenia-spectrum symptoms. During functional MRI (fMRI), a community sample of healthy adults (N = 30) with high and low SA viewed positive, negative, and neutral facial expressions. Afterward, participants completed an online daily diary in which they rated schizophrenia-spectrum symptoms and occurrence of interpersonal conflict each day for 21 days. Compared with low SA, high SA participants had less ventral (V)LPFC activity to positive versus neutral expressions. In addition, participants with a combination of high SA and low VLPFC activity to positive versus neutral expressions had worse daily diary ratings of schizophrenia-spectrum symptoms, including worse cognition, paranoia, motivation/productivity, and vigor/positive affect (i.e., psychomotor activation). Finally, among high SA participants, VLPFC activity predicted the daily relationship between distress from interpersonal conflict and symptom-severity; specifically, high SA participants with low VLPFC activity had worse paranoia on days of high conflict distress. These findings indicate that VLPFC deficits in positive emotion are associated with both SA and other schizophrenia-spectrum symptoms and that understanding the interaction of SA, VLPFC function, and social stress could facilitate the use of SA in the prevention and treatment of schizophrenia.

  9. ARID1A loss correlates with mismatch repair deficiency and intact p53 expression in high-grade endometrial carcinomas

    PubMed Central

    Allo, Ghassan; Bernardini, Marcus Q; Wu, Ren-Chin; Shih, Ie-Ming; Kalloger, Steve; Pollett, Aaron; Gilks, C Blake; Clarke, Blaise A

    2015-01-01

    BAF250a (ARID1A) loss is a frequent event in high-grade endometrial cancers. It has been proposed that ARID1A is a driver gene, with ARID1A mutations occurring secondary to deregulated mismatch repair mechanism in gastric cancers, representing an alternative oncogenic pathway to p53 alteration. The prognostic significance of ARID1A loss is controversial. In this study, we investigated the frequency of BAF250a immunohistochemical loss in a cohort of high-grade endometrial cancers (n = 190) and correlated it with mismatch repair (hMLH1, hMSH2, hMSH6, and hPMS2) and p53 protein expression. The 190 cases consisted of 82 high-grade endometrioid, 88 serous, 10 clear cell, and 10 mixed (carcinosarcomas and mixed histology). There was BAF250a loss in 55/190 (29%) cancers, most commonly in high-grade endometrioid carcinomas (46 vs 9% in serous carcinomas, P<0.0001). Loss of any mismatch repair proteins was observed in 63/190 (33%) cancers, most commonly in high-grade endometrioid carcinomas (57 vs 10% in serous carcinomas, P<0.0001). Aberrant p53 expression was found in 86/190 (45%) cancers, more commonly in serous carcinomas (77 vs 18% in high-grade endometrioid carcinomas, P<0.0001). BAF250a loss was associated with mismatch repair loss (P<0.0001) and normal p53 expression (P<0.0001). These associations were maintained in the subset analysis within the high-grade endometrioid (P = 0.026 and P = 0.0083, respectively) and serous carcinoma cases (P = 0.0031 and P<0.0001, respectively). Survival analysis revealed a superior progression-free survival (P = 0.017) for patients with BAF250a loss within the entire cohort but not within the high-grade endometrioid and serous subtypes. Additionally, data from The Cancer Genome Atlas were extracted to correlate mutations in ARID1A, TP53, and MMR genes; we found that ARID1A mutations were negatively associated with TP53 mutations but were unrelated to mismatch repair gene mutations. In conclusion, BAF250a loss is more common in high

  10. High atomic weight, high-energy radiation (HZE) induces transcriptional responses shared with conventional stresses in addition to a core "DSB" response specific to clastogenic treatments.

    PubMed

    Missirian, Victor; Conklin, Phillip A; Culligan, Kevin M; Huefner, Neil D; Britt, Anne B

    2014-01-01

    Plants exhibit a robust transcriptional response to gamma radiation which includes the induction of transcripts required for homologous recombination and the suppression of transcripts that promote cell cycle progression. Various DNA damaging agents induce different spectra of DNA damage as well as "collateral" damage to other cellular components and therefore are not expected to provoke identical responses by the cell. Here we study the effects of two different types of ionizing radiation (IR) treatment, HZE (1 GeV Fe(26+) high mass, high charge, and high energy relativistic particles) and gamma photons, on the transcriptome of Arabidopsis thaliana seedlings. Both types of IR induce small clusters of radicals that can result in the formation of double strand breaks (DSBs), but HZE also produces linear arrays of extremely clustered damage. We performed these experiments across a range of time points (1.5-24 h after irradiation) in both wild-type plants and in mutants defective in the DSB-sensing protein kinase ATM. The two types of IR exhibit a shared double strand break-repair-related damage response, although they differ slightly in the timing, degree, and ATM-dependence of the response. The ATM-dependent, DNA metabolism-related transcripts of the "DSB response" were also induced by other DNA damaging agents, but were not induced by conventional stresses. Both Gamma and HZE irradiation induced, at 24 h post-irradiation, ATM-dependent transcripts associated with a variety of conventional stresses; these were overrepresented for pathogen response, rather than DNA metabolism. In contrast, only HZE-irradiated plants, at 1.5 h after irradiation, exhibited an additional and very extensive transcriptional response, shared with plants experiencing "extended night." This response was not apparent in gamma-irradiated plants.

  11. 30 CFR 250.807 - Additional requirements for subsurface safety valves and related equipment installed in high...

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... valves and related equipment installed in high pressure high temperature (HPHT) environments. 250.807... pressure high temperature (HPHT) environments. (a) If you plan to install SSSVs and related equipment in an HPHT environment, you must submit detailed information with your Application for Permit to Drill...

  12. A Chemokine Expressed in Lymphoid High Endothelial Venules Promotes the Adhesion and Chemotaxis of Naive T Lymphocytes

    NASA Astrophysics Data System (ADS)

    Gunn, Michael D.; Tangemann, Kirsten; Tam, Carmen; Cyster, Jason G.; Rosen, Steven D.; Williams, Lewis T.

    1998-01-01

    Preferential homing of naive lymphocytes to secondary lymphoid organs is thought to involve the action of chemokines, yet no chemokine has been shown to have either the expression pattern or the activities required to mediate this process. Here we show that a chemokine represented in the EST database, secondary lymphoid-tissue chemokine (SLC), is expressed in the high endothelial venules of lymph nodes and Peyer's patches, in the T cell areas of spleen, lymph nodes, and Peyer's patches, and in the lymphatic endothelium of multiple organs. SLC is a highly efficacious chemoattractant for lymphocytes with preferential activity toward naive T cells. Moreover, SLC induces firm adhesion of naive T lymphocytes via β 2 integrin binding to the counter receptor, intercellular adhesion molecule-1, a necessary step for lymphocyte recruitment. SLC is the first chemokine demonstrated to have the characteristics required to mediate homing of lymphocytes to secondary lymphoid organs. In addition, the expression of SLC in lymphatic endothelium suggests that the migration of lymphocytes from tissues into efferent lymphatics may be an active process mediated by this molecule.

  13. Transgenic maize plants expressing the Totivirus antifungal protein, KP4, are highly resistant to corn smut.

    PubMed

    Allen, Aron; Islamovic, Emir; Kaur, Jagdeep; Gold, Scott; Shah, Dilip; Smith, Thomas J

    2011-10-01

    The corn smut fungus, Ustilago maydis, is a global pathogen responsible for extensive agricultural losses. Control of corn smut using traditional breeding has met with limited success because natural resistance to U. maydis is organ specific and involves numerous maize genes. Here, we present a transgenic approach by constitutively expressing the Totivirus antifungal protein KP4, in maize. Transgenic maize plants expressed high levels of KP4 with no apparent negative impact on plant development and displayed robust resistance to U. maydis challenges to both the stem and ear tissues in the greenhouse. More broadly, these results demonstrate that a high level of organ independent fungal resistance can be afforded by transgenic expression of this family of antifungal proteins.

  14. Rapid selection using G418 of high copy number transformants of Pichia pastoris for high-level foreign gene expression.

    PubMed

    Scorer, C A; Clare, J J; McCombie, W R; Romanos, M A; Sreekrishna, K

    1994-02-01

    Pichia pastoris is a methylotrophic yeast increasingly important in the production of therapeutic proteins. Expression vectors are based on the methanol-inducible AOX1 promoter and are integrated into the host chromosome. In most cases high copy number integration has been shown to be important for high-level expression. Since this occurs at low frequency during transformation, we previously used DNA dot blot screens to identify suitable clones. In this paper we report the use of vectors containing the Tn903 kanr gene conferring G418-resistance. Initial experiments demonstrated that copy number showed a tight correlation with drug-resistance. Using a G418 growth inhibition screen, we readily isolated a series of transformants, containing progressively increasing numbers (1 to 12) of a vector expressing HIV-1 ENV, which we used to examine the relationship between copy number and foreign mRNA levels. Northern blot analysis indicated that ENV mRNA levels from a single-copy clone were nearly as high as AOX1 mRNA, and increased progressively with increasing copy number so as to greatly exceed AOX1 mRNA. We have also developed protocols for the selection, using G418, of high copy number transformants following spheroplast transformation or electroporation. We anticipate that these protocols will simplify the use of Pichia as a biotechnological tool.

  15. High energy particles at Mars and Venus: Phobos-2, Mars Express and Venus Express observations and their interpretation by hybrid model simulations

    NASA Astrophysics Data System (ADS)

    McKenna-Lawlor, Susan; Kallio, Esa; Fram, Rudy A.; Alho, Markku; Jarvinen, Riku; Dyadechkin, Sergey; Wedlund, Cyril Simon; Zhang, Tielong; Collinson, Glyn A.; Futaana, Yoshifumi

    2013-04-01

    Mars and Venus can both be reached by Solar Energetic Particles (SEPs). Such high energy particles (protons, multiply charged heavy ions, electrons) penetrate the upper atmospheres of Mars and Venus because, in contrast to Earth, these bodies do not have a significant, global, intrinsic magnetic field to exclude them. One especially well documented, complex and prolonged SEP took in place in early 1989 (Solar Cycle 23) when the Phobos-2 spacecraft was orbiting Mars. This spacecraft had a dedicated high energy particle instrument onboard (SLED), which measured particles with energies in the keV range up to a few tens of MeV. There was in addition a magnetometer as well as solar wind plasma detectors onboard which together provided complementary data to support contemporaneous studies of the background SEP environment. Currently, while the Sun is displaying maximum activity (Solar Cycle 24), Mars and Venus are being individually monitored by instrumentation flown onboard the Mars Express (MEX) and Venus Express (VEX) spacecraft. Neither of these spacecraft carry a high energy particle instrument but their Analyzer of Space Plasmas and Energetic Atoms (ASPERA) experiments (ASPERA-3 on MEX and ASPERA-4 on VEX), can be used to study SEPs integrated over E ≥ ~30 MeV which penetrate the instrument hardware and form background counts in the plasma data. In the present work we present SEP events measured at Mars and Venus based on Phobos-2, 1989 data and on, more recent, MEX and VEX (identified from particle background) observations. We further introduce numerical global SEP simulations of the measured events based on 3-D self-consistent hybrid models (HYB-Mars and HYB-Venus). Through comparing the in situ SEP observations with these simulations, new insights are provided into the properties of the measured SEPs as well as into how their individual planetary bow shocks and magnetospheres affect the characteristics of their ambient Martian and Venusian SEP environments.

  16. Squaramide-tertiary amine catalyzed asymmetric cascade sulfa-Michael/Michael addition via dynamic kinetic resolution: access to highly functionalized chromans with three contiguous stereocenters.

    PubMed

    Yang, Wen; Yang, Yi; Du, Da-Ming

    2013-03-15

    An efficient asymmetric cascade sulfa-Michael/Michael addition reaction catalyzed by a chiral bifunctional squaramide-tertiary amine catalyst has been developed. This organocatalytic cascade reaction provides easy access to highly functionalized chromans with three contiguous stereocenters, including one quaternary center. In addition, a novel cascade sulfa Michael/retro-sulfa-Michael/sulfa-Michael/Michael reaction process, involving dynamic kinetic resolution, is described.

  17. Gene expression profiling in undifferentiated thyroid carcinoma induced by high-dose radiation

    PubMed Central

    Bang, Hyun Soon; Choi, Moo Hyun; Kim, Cha Soon; Choi, Seung Jin

    2016-01-01

    Published gene expression studies for radiation-induced thyroid carcinogenesis have used various methodologies. In this study, we identified differential gene expression in a human thyroid epithelial cell line after exposure to high-dose γ-radiation. HTori-3 cells were exposed to 5 or 10 Gy of ionizing radiation using two dose rates (high-dose rate: 4.68 Gy/min, and low-dose rate: 40 mGy/h) and then implanted into the backs of BALB/c nude mice after 4 (10 Gy) or 5 weeks (5 Gy). Decreases in cell viability, increases in giant cell frequency, anchorage-independent growth in vitro, and tumorigenicity in vivo were observed. Particularly, the cells irradiated with 5 Gy at the high-dose rate or 10 Gy at the low-dose rate demonstrated more prominent tumorigenicity. Gene expression profiling was analyzed via microarray. Numerous genes that were significantly altered by a fold-change of >50% following irradiation were identified in each group. Gene expression analysis identified six commonly misregulated genes, including CRYAB, IL-18, ZNF845, CYP24A1, OR4N4 and VN1R4, at all doses. These genes involve apoptosis, the immune response, regulation of transcription, and receptor signaling pathways. Overall, the altered genes in high-dose rate (HDR) 5 Gy and low-dose rate (LDR) 10 Gy were more than those of LDR 5 Gy and HDR 10 Gy. Thus, we investigated genes associated with aggressive tumor development using the two dosage treatments. In this study, the identified gene expression profiles reflect the molecular response following high doses of external radiation exposure and may provide helpful information about radiation-induced thyroid tumors in the high-dose range. PMID:27006382

  18. A High-content Imaging Workflow to Study Grb2 Signaling Complexes by Expression Cloning

    PubMed Central

    Freeman, Jamie; Kriston-Vizi, Janos; Seed, Brian; Ketteler, Robin

    2012-01-01

    Signal transduction by growth factor receptors is essential for cells to maintain proliferation and differentiation and requires tight control. Signal transduction is initiated by binding of an external ligand to a transmembrane receptor and activation of downstream signaling cascades. A key regulator of mitogenic signaling is Grb2, a modular protein composed of an internal SH2 (Src Homology 2) domain flanked by two SH3 domains that lacks enzymatic activity. Grb2 is constitutively associated with the GTPase Son-Of-Sevenless (SOS) via its N-terminal SH3 domain. The SH2 domain of Grb2 binds to growth factor receptors at phosphorylated tyrosine residues thus coupling receptor activation to the SOS-Ras-MAP kinase signaling cascade. In addition, other roles for Grb2 as a positive or negative regulator of signaling and receptor endocytosis have been described. The modular composition of Grb2 suggests that it can dock to a variety of receptors and transduce signals along a multitude of different pathways1-3. Described here is a simple microscopy assay that monitors recruitment of Grb2 to the plasma membrane. It is adapted from an assay that measures changes in sub-cellular localization of green-fluorescent protein (GFP)-tagged Grb2 in response to a stimulus4-6. Plasma membrane receptors that bind Grb2 such as activated Epidermal Growth Factor Receptor (EGFR) recruit GFP-Grb2 to the plasma membrane upon cDNA expression and subsequently relocate to endosomal compartments in the cell. In order to identify in vivo protein complexes of Grb2, this technique can be used to perform a genome-wide high-content screen based on changes in Grb2 sub-cellular localization. The preparation of cDNA expression clones, transfection and image acquisition are described in detail below. Compared to other genomic methods used to identify protein interaction partners, such as yeast-two-hybrid, this technique allows the visualization of protein complexes in mammalian cells at the sub

  19. Insect cell transformation vectors that support high level expression and promoter assessment in insect cell culture

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A somatic transformation vector, pDP9, was constructed that provides a simplified means of producing permanently transformed cultured insect cells that support high levels of protein expression of foreign genes. The pDP9 plasmid vector incorporates DNA sequences from the Junonia coenia densovirus th...

  20. Plastid-expressed 5-enolpyruvylshikimate-3-phosphate synthase genes provide high level glyphosate tolerance in tobacco.

    PubMed

    Ye, G N; Hajdukiewicz, P T; Broyles, D; Rodriguez, D; Xu, C W; Nehra, N; Staub, J M

    2001-02-01

    Plastid transformation (transplastomic) technology has several potential advantages for biotechnological applications including the use of unmodified prokaryotic genes for engineering, potential high-level gene expression and gene containment due to maternal inheritance in most crop plants. However, the efficacy of a plastid-encoded trait may change depending on plastid number and tissue type. We report a feasibility study in tobacco plastids to achieve high-level herbicide resistance in both vegetative tissues and reproductive organs. We chose to test glyphosate resistance via over-expression in plastids of tolerant forms of 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS). Immunological, enzymatic and whole-plant assays were used to prove the efficacy of three different prokaryotic (Achromobacter, Agrobacterium and Bacillus) EPSPS genes. Using the Agrobacterium strain CP4 EPSPS as a model we identified translational control sequences that direct a 10,000-fold range of protein accumulation (to >10% total soluble protein in leaves). Plastid-expressed EPSPS could provide very high levels of glyphosate resistance, although levels of resistance in vegetative and reproductive tissues differed depending on EPSPS accumulation levels, and correlated to the plastid abundance in these tissues. Paradoxically, higher levels of plastid-expressed EPSPS protein accumulation were apparently required for efficacy than from a similar nuclear-encoded gene. Nevertheless, the demonstration of high-level glyphosate tolerance in vegetative and reproductive organs using transplastomic technology provides a necessary step for transfer of this technology to other crop species.

  1. High-level expression and preparation of recombinant human fibrinogen as biopharmaceuticals.

    PubMed

    Hirashima, Masaki; Imamura, Takayuki; Yano, Kentaro; Kawamura, Ryoichi; Meta, Akihiro; Tokieda, Yoshiyuki; Nakashima, Toshihiro

    2016-02-01

    Fibrinogen is a large and complex glycoprotein containing two sets of each of three different chains (α, β and γ). There have been no reports of high-level expression of fibrinogen at commercial levels using mammalian cultured cells such as CHO cells because of the difficulty in highly expressing a protein with such a complex structure. We achieved high-level (1.3 g/l or higher) expression of recombinant human fibrinogen using CHO DG44 cells by optimizing the expression system and culture conditions. We also succeeded in establishing a high-recovery preparation method for recombinant fibrinogen that rarely yields degraded products. To characterize the properties of the recombinant human fibrinogen, we performed SDS-PAGE; western blotting of the α, β and γ chains using specific antibodies and scanning electron microscopy observations of fibrin fibres. We also evaluated the functional equivalence between recombinant fibrinogen and plasma fibrinogen with respect to the release of fibrinopeptides initiated by thrombin and its cross-linking properties. The basic properties of recombinant fibrinogen showed no apparent differences from those of plasma fibrinogen. Here, we report the development of methods for the culture and preparation of recombinant human fibrinogen of satisfactory quality that can be scaled up to the commercial level.

  2. Social Skills Expression of Senior High School Age Students in Physical Education Classes

    ERIC Educational Resources Information Center

    Akelaitis, Arturas V.

    2015-01-01

    The main purpose of the present study is to reveal the peculiarities of social skills expression of senior high school age students in physical education classes. The independent random sample consisted of 244 (15-16 years old) students and 258 (17-18 years old) students, of which there were 224 boys and 278 girls. L. Bulotaite and V. Gudžinskiene…

  3. Emotional Experience, Expression, and Regulation of High-Quality Japanese Elementary School Teachers

    ERIC Educational Resources Information Center

    Hosotani, Rika; Imai-Matsumura, Kyoko

    2011-01-01

    The present study investigates the emotional experience, expression, and regulation processes of high-quality Japanese elementary school teachers while they interact with children, in terms of teachers' emotional competence. Qualitative analysis of interview data demonstrated that teachers had various emotional experiences including self-elicited…

  4. High-level production of a functional immunoglobulin heterodimer in a baculovirus expression system.

    PubMed Central

    Hasemann, C A; Capra, J D

    1990-01-01

    A murine immunoglobulin heterodimer has been expressed in a baculovirus expression system. This was achieved by using both double infection of insect cells with separate heavy- and light-chain-expressing viruses and infection with a double-recombinant virus containing both the immunoglobulin heavy- and light-chain cDNAs. In both cases, the polypeptide chains were correctly processed, glycosylated, and assembled into normal H2L2 (H = heavy, L = light) immunoglobulin monomers. These molecules bound antigen and expressed both polyclonal idiotype and monoclonal idiotopes. Furthermore, the transfer vectors described have been modified to contain the F1 origin of replication for the production of single-stranded DNA, which facilitates site-specific mutations of either the polyhedrin promoter or the inserted foreign gene. Use of this system should significantly advance the analysis of the structural bases for both idiotype expression and antigen binding by immunoglobulin. More importantly, it provides a generic method for the high-level expression of antibodies of diverse interest. Images PMID:2111022

  5. TSLP Expression and High Serum TSLP Level Indicate a Poor Prognosis in Gastric Cancer Patients

    PubMed Central

    Watanabe, Joji; Saito, Hiroaki; Miyatani, Kozo; Ikeguchi, Masahide; Umekita, Yoshihisa

    2015-01-01

    Background Thymic stromal lymphopoietin (TSLP) plays an important role in promoting tumor survival, by manipulating the immune response and angiogenesis. However, the clinical significance of TSLP in gastric cancer is unclear. Methods Immunohistochemistry was used to investigate TSLP expression in non-cancerous gastric mucosa and gastric cancer tissue from patients with gastric cancer. Serum TSLP levels were measured using an enzyme-linked immunosorbent assay. Results Tumors with TSLP expression were significantly larger than those without TSLP expression. TSLP expression was observed more frequently in advanced (T2/T3/T4) than in early (T1) gastric cancer and in stage 3/4 than in stage 1/2. Lymph node metastasis, liver metastasis, positive peritoneal lavage cytology, lymphatic invasion, and vascular invasion occurred significantly more often in TSLP-expressing than in non-expressing tumors. The prognosis of patients with TSLP-positive tumors was significantly worse than that of patients with TSLP-negative tumors. Patients with high serum TSLP concentrations also had a significantly worse prognosis than those with low concentrations. Multivariate analysis identified serum TSLP level as an independent prognostic indicator. Conclusion TSLP is closely related to the progression of gastric cancer and may predict survival in these patients. PMID:26538800

  6. The infectious BAC genomic DNA expression library: a high capacity vector system for functional genomics

    PubMed Central

    Lufino, Michele M. P.; Edser, Pauline A. H.; Quail, Michael A.; Rice, Stephen; Adams, David J.; Wade-Martins, Richard

    2016-01-01

    Gene dosage plays a critical role in a range of cellular phenotypes, yet most cellular expression systems use heterologous cDNA-based vectors which express proteins well above physiological levels. In contrast, genomic DNA expression vectors generate physiologically-relevant levels of gene expression by carrying the whole genomic DNA locus of a gene including its regulatory elements. Here we describe the first genomic DNA expression library generated using the high-capacity herpes simplex virus-1 amplicon technology to deliver bacterial artificial chromosomes (BACs) into cells by viral transduction. The infectious BAC (iBAC) library contains 184,320 clones with an average insert size of 134.5 kb. We show in a Chinese hamster ovary (CHO) disease model cell line and mouse embryonic stem (ES) cells that this library can be used for genetic rescue studies in a range of contexts including the physiological restoration of Ldlr deficiency, and viral receptor expression. The iBAC library represents an important new genetic analysis tool openly available to the research community. PMID:27353647

  7. High expression of TACC3 in esophageal squamous cell carcinoma correlates with poor prognosis.

    PubMed

    Huang, Zhi-Liang; Lin, Zhi-Rui; Xiao, Yi-Ren; Cao, Xun; Zhu, Lin-Chun; Zeng, Mu-Sheng; Zhong, Qian; Wen, Zhe-Sheng

    2015-03-30

    To analyze the expression of the transforming acidic coiled-coil protein 3 (TACC3) in esophageal squamous cell carcinoma (ESCC) samples, and to identify whether TACC3 can serve as a biomarker for the diagnosis and prognosis of ESCC, qPCR, western blotting and immunohistochemistry staining (IHC) were utilized to detect the expression of TACC3. Furthermore, cell growth, colony formation, migration ability and the epithelial-mesenchymal transition markers of ESCC cells in which TACC3 were knocked-down were measured. The mRNA and protein levels of TACC3 were higher in ESCC specimens compared to non-tumorous esophageal epithelial tissues. IHC results revealed TACC3 expression was significantly correlated to differentiation (p = 0.017) and lymphoid nodal status (p = 0.028). The patients with high-expression of TACC3 had a significantly poor prognosis compared to those of low-expression (p = 0.017), especially in the patients at stages I-II (p = 0.028). Multivariate analysis indicated that TACC3 expression was an independent prognostic factor for ESCC patients (p = 0.025). Knockdown of TACC3 inhibited the ability of cell proliferation, colony formation and migration. This study first identifies TACC3 not only as a useful biomarker for diagnose and prognosis of ESCC, but also as a potential therapeutic target for patients with ESCC.

  8. High-level expression, purification and antibacterial activity of bovine lactoferricin and lactoferrampin in Photorhabdus luminescens.

    PubMed

    Tang, Zhiru; Zhang, Youming; Stewart, Adrian Francis; Geng, Meimei; Tang, Xiangsha; Tu, Qiang; Yin, Yulong

    2010-10-01

    Bovine lactoferricin (LFC) and bovine lactoferrampin (LFA) are two active fragments located in the N(1)-domain of bovine lactoferrin. Recent studies suggested that LFC and LFA have broad-spectrum activity against Gram-positive and Gram-negative bacteria. To date, LFC and LFA have usually been produced from milk. We report here the high-level expression, purification and characterization of LFC and LFA using the Photorhabdus luminescens expression system. After the cipA and cipB genes were deleted by ET recombination, the expression host P. luminescens TZR(001) was constructed. A synthetic LFC-LFA gene containing LFC and LFA was fused with the cipB gene to form a cipB-LFC-LFA gene. To obtain the expression vector pBAD-cipB-LFC-LFA, the cipB-LFC-LFA gene was cloned on the L-arabinose-inducible expression vector pBAD24. pBAD-cipB-LFC-LFA was transformed into P. luminescens TZR(001). The cipB-LFC-LFA fusion protein was expressed under the induction of L-arabinose and its yield reached 12 mg L(-1) bacterial culture. Recombinant LFC-LFA was released from cipB by pepsin. The MIC of recombinant LFC-LFA toward E. coli 0149, 0141 and 020 was 6.25, 12.5 and 3.175 microg ml(-1), respectively. PMID:20510367

  9. Comparative gene expression profiles for highly similar aggressive phenotypes in male and female cichlid fishes (Julidochromis)

    PubMed Central

    Schumer, Molly; Krishnakant, Kavita; Renn, Suzy C. P.

    2011-01-01

    SUMMARY Julidochromis marlieri and Julidochromis transcriptus are two closely related Tanganyikan cichlids that have evolved different behavior and mating strategies since they diverged from their common ancestor. While J. transcriptus follows the ancestral pattern of male dominance, male-biased sexual size dimorphism and territoriality, the pattern is reversed in J. marlieri. In J. marlieri, females show all of these behavioral and morphological characteristics. This raises the question of whether female J. marlieri achieve the dominant phenotype by expressing the same genes as J. transcriptus males or whether novel brain gene expression patterns have evolved to produce a similar behavioral phenotype in the females of J. marlieri. This study used cDNA microarrays to investigate whether female J. marlieri and male J. transcriptus show conserved or divergent patterns of brain gene expression. Analysis of microarray data in both species showed certain gene expression patterns associated with sex role independent of gonadal sex and, to a lesser extent, gene expression patterns associated with sex independent of sex role. In general, these data suggest that while there has been substantial divergence in gene expression patterns between J. transcriptus and J. marlieri, we can detect a highly significant overlap for a core set of genes related to aggression in both species. These results suggest that the proximate mechanisms regulating aggressive behavior in J. transcriptus and J. marlieri may be shared. PMID:21900474

  10. Profiling status epilepticus-induced changes in hippocampal RNA expression using high-throughput RNA sequencing

    PubMed Central

    Hansen, Katelin F.; Sakamoto, Kensuke; Pelz, Carl; Impey, Soren; Obrietan, Karl

    2014-01-01

    Status epilepticus (SE) is a life-threatening condition that can give rise to a number of neurological disorders, including learning deficits, depression, and epilepsy. Many of the effects of SE appear to be mediated by alterations in gene expression. To gain deeper insight into how SE affects the transcriptome, we employed the pilocarpine SE model in mice and Illumina-based high-throughput sequencing to characterize alterations in gene expression from the induction of SE, to the development of spontaneous seizure activity. While some genes were upregulated over the entire course of the pathological progression, each of the three sequenced time points (12-hour, 10-days and 6-weeks post-SE) had a largely unique transcriptional profile. Hence, genes that regulate synaptic physiology and transcription were most prominently altered at 12-hours post-SE; at 10-days post-SE, marked changes in metabolic and homeostatic gene expression were detected; at 6-weeks, substantial changes in the expression of cell excitability and morphogenesis genes were detected. At the level of cell signaling, KEGG analysis revealed dynamic changes within the MAPK pathways, as well as in CREB-associated gene expression. Notably, the inducible expression of several noncoding transcripts was also detected. These findings offer potential new insights into the cellular events that shape SE-evoked pathology. PMID:25373493

  11. Profiling status epilepticus-induced changes in hippocampal RNA expression using high-throughput RNA sequencing.

    PubMed

    Hansen, Katelin F; Sakamoto, Kensuke; Pelz, Carl; Impey, Soren; Obrietan, Karl

    2014-01-01

    Status epilepticus (SE) is a life-threatening condition that can give rise to a number of neurological disorders, including learning deficits, depression, and epilepsy. Many of the effects of SE appear to be mediated by alterations in gene expression. To gain deeper insight into how SE affects the transcriptome, we employed the pilocarpine SE model in mice and Illumina-based high-throughput sequencing to characterize alterations in gene expression from the induction of SE, to the development of spontaneous seizure activity. While some genes were upregulated over the entire course of the pathological progression, each of the three sequenced time points (12-hour, 10-days and 6-weeks post-SE) had a largely unique transcriptional profile. Hence, genes that regulate synaptic physiology and transcription were most prominently altered at 12-hours post-SE; at 10-days post-SE, marked changes in metabolic and homeostatic gene expression were detected; at 6-weeks, substantial changes in the expression of cell excitability and morphogenesis genes were detected. At the level of cell signaling, KEGG analysis revealed dynamic changes within the MAPK pathways, as well as in CREB-associated gene expression. Notably, the inducible expression of several noncoding transcripts was also detected. These findings offer potential new insights into the cellular events that shape SE-evoked pathology. PMID:25373493

  12. Targeted toxin-based selectable drug-free enrichment of Mammalian cells with high transgene expression.

    PubMed

    Sato, Masahiro; Akasaka, Eri; Saitoh, Issei; Ohtsuka, Masato; Nakamura, Shingo; Sakurai, Takayuki; Watanabe, Satoshi

    2013-01-01

    Almost all transfection protocols for mammalian cells use a drug resistance gene for the selection of transfected cells. However, it always requires the characterization of each isolated clone regarding transgene expression, which is time-consuming and labor-intensive. In the current study, we developed a novel method to selectively isolate clones with high transgene expression without drug selection. Porcine embryonic fibroblasts were transfected with pCEIEnd, an expression vector that simultaneously expresses enhanced green fluorescent protein (EGFP) and endo-b-galactosidase C(EndoGalC; an enzyme capable of digesting cell surface a-Gal epitope) upon transfection. After transfection, the surviving cells were briefly treated with IB4SAP (a-Gal epitope-specific BS-I-B4 lectin conjugated with a toxin saporin). The treated cells were then allowed to grow in normal medium, during which only cells strongly expressing EndoGalC and EGFP would survive because of the absence of a-Gal epitopes on their cell surface. Almost all the surviving colonies after IB4SAP treatment were in fact negative for BS-I-B4 staining, and also strongly expressed EGFP. This system would be particularly valuable for researchers who wish to perform large-scale production of therapeutically important recombinant proteins. PMID:24832665

  13. Single Nucleotide Polymorphisms in Regulator-Encoding Genes Have an Additive Effect on Virulence Gene Expression in a Vibrio cholerae Clinical Isolate.

    PubMed

    Carignan, Bailey M; Brumfield, Kyle D; Son, Mike S

    2016-01-01

    Vibrio cholerae is the etiological agent of the infectious disease cholera, which is characterized by vomiting and severe watery diarrhea. Recently, V. cholerae clinical isolates have demonstrated increased virulence capabilities, causing more severe symptoms with a much higher rate of disease progression than previously observed. We have identified single nucleotide polymorphisms (SNPs) in four virulence-regulatory genes (hapR, hns, luxO, and vieA) of a hypervirulent V. cholerae clinical isolate, MQ1795. Herein, all SNPs and SNP combinations of interest were introduced into the prototypical El Tor reference strain N16961, and the effects on the production of numerous virulence-related factors, including cholera toxin (CT), the toxin-coregulated pilus (TCP), and ToxT, were analyzed. Our data show that triple-SNP (hapR hns luxO and hns luxO vieA) and quadruple-SNP combinations produced the greatest increases in CT, TCP, and ToxT production. The hns and hns luxO SNP combinations were sufficient for increased TCP and ToxT production. Notably, the hns luxO vieA triple-SNP combination strain produced TCP and ToxT levels similar to those of MQ1795. Certain SNP combinations (hapR and hapR vieA) had the opposite effect on CT, TCP, and ToxT expression. Interestingly, the hns vieA double-SNP combination strain increased TCP production while decreasing CT production. Our findings suggest that SNPs identified in the four regulatory genes, in various combinations, are associated with increased virulence capabilities observed in V. cholerae clinical isolates. These studies provide insight into the evolution of highly virulent strains. IMPORTANCE Cholera, an infectious disease of the small intestine caused by the aquatic bacterium Vibrio cholerae, often results in vomiting and acute watery diarrhea. If left untreated or if the response is too slow, the symptoms can quickly lead to extreme dehydration and ultimately death of the patient. Recent anecdotal evidence of cholera

  14. Single Nucleotide Polymorphisms in Regulator-Encoding Genes Have an Additive Effect on Virulence Gene Expression in a Vibrio cholerae Clinical Isolate

    PubMed Central

    Carignan, Bailey M.; Brumfield, Kyle D.

    2016-01-01

    ABSTRACT Vibrio cholerae is the etiological agent of the infectious disease cholera, which is characterized by vomiting and severe watery diarrhea. Recently, V. cholerae clinical isolates have demonstrated increased virulence capabilities, causing more severe symptoms with a much higher rate of disease progression than previously observed. We have identified single nucleotide polymorphisms (SNPs) in four virulence-regulatory genes (hapR, hns, luxO, and vieA) of a hypervirulent V. cholerae clinical isolate, MQ1795. Herein, all SNPs and SNP combinations of interest were introduced into the prototypical El Tor reference strain N16961, and the effects on the production of numerous virulence-related factors, including cholera toxin (CT), the toxin-coregulated pilus (TCP), and ToxT, were analyzed. Our data show that triple-SNP (hapR hns luxO and hns luxO vieA) and quadruple-SNP combinations produced the greatest increases in CT, TCP, and ToxT production. The hns and hns luxO SNP combinations were sufficient for increased TCP and ToxT production. Notably, the hns luxO vieA triple-SNP combination strain produced TCP and ToxT levels similar to those of MQ1795. Certain SNP combinations (hapR and hapR vieA) had the opposite effect on CT, TCP, and ToxT expression. Interestingly, the hns vieA double-SNP combination strain increased TCP production while decreasing CT production. Our findings suggest that SNPs identified in the four regulatory genes, in various combinations, are associated with increased virulence capabilities observed in V. cholerae clinical isolates. These studies provide insight into the evolution of highly virulent strains. IMPORTANCE Cholera, an infectious disease of the small intestine caused by the aquatic bacterium Vibrio cholerae, often results in vomiting and acute watery diarrhea. If left untreated or if the response is too slow, the symptoms can quickly lead to extreme dehydration and ultimately death of the patient. Recent anecdotal evidence of

  15. Single Nucleotide Polymorphisms in Regulator-Encoding Genes Have an Additive Effect on Virulence Gene Expression in a Vibrio cholerae Clinical Isolate

    PubMed Central

    Carignan, Bailey M.; Brumfield, Kyle D.

    2016-01-01

    ABSTRACT Vibrio cholerae is the etiological agent of the infectious disease cholera, which is characterized by vomiting and severe watery diarrhea. Recently, V. cholerae clinical isolates have demonstrated increased virulence capabilities, causing more severe symptoms with a much higher rate of disease progression than previously observed. We have identified single nucleotide polymorphisms (SNPs) in four virulence-regulatory genes (hapR, hns, luxO, and vieA) of a hypervirulent V. cholerae clinical isolate, MQ1795. Herein, all SNPs and SNP combinations of interest were introduced into the prototypical El Tor reference strain N16961, and the effects on the production of numerous virulence-related factors, including cholera toxin (CT), the toxin-coregulated pilus (TCP), and ToxT, were analyzed. Our data show that triple-SNP (hapR hns luxO and hns luxO vieA) and quadruple-SNP combinations produced the greatest increases in CT, TCP, and ToxT production. The hns and hns luxO SNP combinations were sufficient for increased TCP and ToxT production. Notably, the hns luxO vieA triple-SNP combination strain produced TCP and ToxT levels similar to those of MQ1795. Certain SNP combinations (hapR and hapR vieA) had the opposite effect on CT, TCP, and ToxT expression. Interestingly, the hns vieA double-SNP combination strain increased TCP production while decreasing CT production. Our findings suggest that SNPs identified in the four regulatory genes, in various combinations, are associated with increased virulence capabilities observed in V. cholerae clinical isolates. These studies provide insight into the evolution of highly virulent strains. IMPORTANCE Cholera, an infectious disease of the small intestine caused by the aquatic bacterium Vibrio cholerae, often results in vomiting and acute watery diarrhea. If left untreated or if the response is too slow, the symptoms can quickly lead to extreme dehydration and ultimately death of the patient. Recent anecdotal evidence of

  16. High expression of WISP-1 correlates with poor prognosis in pancreatic ductal adenocarcinoma.

    PubMed

    Yang, Jian-Yu; Yang, Min-Wei; Huo, Yan-Miao; Liu, Wei; Liu, De-Jun; Li, Jiao; Zhang, Jun-Feng; Hua, Rong; Sun, Yong-Wei

    2015-01-01

    WNT1 inducible signaling pathway protein 1 (WISP-1) is a member of the CCN family of growth factors and reported to possess an important role in tumorigenesis by triggering downstream events via integrin signaling. However, the exact role of WISP-1 in cancer remains unclear. In this study, we examined the expression pattern of WISP-1 at both mRNA and protein levels and evaluated the prognostic value of WISP-1 in pancreatic ductal adenocarcinoma (PDA). Expression of WISP-1 at mRNA level was upregulated in 17/24 tumor tissues compared to the matched adjacent non-tumor tissues and the result was confirmed by western blotting at protein level. Immunohistochemical staining of 194 pairs of PDA specimens suggested that high expression of WISP-1 is strongly correlated with clinical stage (P=0.003), T classification (P=0.008) and liver metastasis (P=0.012). Consistently, Kaplan-Meier survival curves indicated that patients with high expression of WISP-1 had a shorter survival time independent of clinical stage and lymphatic metastasis status. Moreover, univariate and multivariate analysis confirmed WISP-1 expression, age, classification and liver metastasis as independent prognostic factors for overall survival of PDA patients. Taken together, these results suggest that WISP-1 may serve as a potential prognostic biomarker for PDA.

  17. IRES-mediated Tricistronic vectors for enhancing generation of high monoclonal antibody expressing CHO cell lines.

    PubMed

    Ho, Steven C L; Bardor, Muriel; Feng, Huatao; Mariati; Tong, Yen Wah; Song, Zhiwei; Yap, Miranda G S; Yang, Yuansheng

    2012-01-01

    A Tricistronic vector utilizing internal ribosome entry site (IRES) elements to express the light chain (LC), heavy chain (HC), and a neomycin phosphotransferase (NPT) selection marker from one transcript is designed for generation of mAb expressing CHO cell lines. As compared to the commonly used vectors, benefits of this design include: (1) minimized non-expressing clones, (2) enhanced stable mAb productivity without gene amplification, (3) control of LC and HC expression at defined ratios, and (4) consistent product quality. After optimization of the LC and HC arrangement and increasing selection stringency by weakening the NPT selection marker, this Tricistronic vector is able to generate stably transfected pools with specific productivity (qmAb) greater than 5pg/cell/day (pcd) and titers over 150mg/L. 5% of clones from these pools have qmAb greater than 20pcd and titers ranging from 300 to more than 500mg/L under non-optimized shake flask batch cultures using commercially available protein-free medium. The mAb produced by these clones have low aggregation and consistent glycosylation profiles. The entire process of transfection to high-expressing clones requires only 6 months. The IRES-mediated Tricistronic vector provides an attractive alternative to commonly used vectors for fast generation of mAb CHO cell lines with high productivity. PMID:22024589

  18. Localized brain differences in Arc expression between mice showing low vs. high propensity to ethanol sensitization.

    PubMed

    Nona, Christina N; Lam, Marcus; Nobrega, José N

    2016-03-01

    Behavioral sensitization to ethanol (EtOH) manifests as a progressive and enduring increase in locomotor activity with repeated drug exposure. However, not all mice sensitize to EtOH and the neuronal mechanisms mediating vulnerability and resistance to EtOH sensitization remain unclear. We examined regional brain expression of the immediate early gene activity-regulated cytoskeleton-associated protein (Arc) in order to identify brain areas in which neuroplastic changes may contribute to the development and expression of EtOH sensitization. Male DBA/2J mice received 5 biweekly injections of EtOH (2.2g/kg, i.p.) or saline (SAL). They were categorized as high- (HS) or low-sensitized (LS) on the basis of final locomotor activity scores. In both LS and HS mice sacrificed after the last sensitization injection, Arc expression was decreased throughout the brain in comparison to SAL animals. A similar pattern was seen in mice sacrificed after an EtOH challenge two weeks after the last sensitization injection. However in this cohort, Arc expression was significantly increased in the central amygdala (CeA) in LS mice and in SAL mice receiving EtOH for the first time. No significant increases in Arc expression were seen in brains of sensitized (HS) animals. These results indicate an acute EtOH challenge results in different patterns of Arc expression in brains of LS, HS, and SAL mice. The dramatic increases in Arc expression in the CeA in LS and SAL mice showing little or no behavioral activation suggests that neural activity in this region may serve to inhibit the stimulant effects of EtOH. The observation that HS mice do not show increases in Arc expression with an EtOH challenge suggests the possibility that increased tolerance to the Arc-inducing effects of EtOH may be a factor in behavioral sensitization.

  19. Implementing an Additive, College Access and Readiness Program for Latina/o High School Students in the U.S.

    ERIC Educational Resources Information Center

    Amaro-Jimenez, Carla; Hungerford-Kresser, Holly

    2013-01-01

    In this article we draw on the experiences of a diverse group of 34 first-generation college students, collected over a year, who served as peer mentors to minority and Latina/o high school students enrolled in four Title I (low-income) high schools in the Southwest U.S. The article identifies the successes and challenges of implementing an…

  20. A Comprehensive Analysis of the Combined Effects of High Light and High Temperature Stresses on Gene Expression in Sunflower

    PubMed Central

    Hewezi, Tarek; Léger, Mathieu; Gentzbittel, Laurent

    2008-01-01

    Background and Aims Although high light (HL) and high temperature (HT) stresses have been extensively investigated, a global analysis of their combined effects on the transcriptome of any plant species has not yet been described. Sunflower is an agronomically important oil crop frequently subjected to these stress factors. Because results in model plants may not always translate well to crop plants, responses of sunflower (Helianthus annuus) to HL, HT and a combination of both stresses were analysed by profiling gene expression in leaves and immature seeds. Methods Plants were grown in HL (600 µE m−2 s−1), HT (35 °C) and a combination of HL and HT (HL + HT), and gene expression in leaves and immature seeds was profiled using cDNA microarrays containing more than 8000 putative unigenes. Key Results Using two-way analysis of variance, 105, 55 and 129 cDNA clones were identified showing significant changes in steady-state transcript levels, across the two tissues, in response to HL, HT and HL + HT, respectively. A significant number of these transcripts were found to be specific to each stress. Comparing gene expression profiles between leaves and immature seeds revealed that 89, 113 and 186 cDNA clones can be considered as differentially expressed in response to HL, HT and HL + HT, respectively. More than half of the cDNA clones showing significant differences between embryo and leaf tissues in response to HL + HT were specific to this stress. Significant differences between leaves and seeds shared by all three stress treatments were observed for only eight genes. Conclusions Taken together, these results indicate that vegetative and reproductive tissues employ different transcriptome responses to these stress treatments. Careful examination of the putative functions of these genes revealed novel and specific responses. The potential roles of many of the differentially expressed genes in stress tolerance are mentioned and discussed. PMID:18477560

  1. [Simultaneous rapid determination of eight food additives in foods by reversed-phase high performance liquid chromatography].

    PubMed

    Zhang, X Y

    2000-11-01

    A simple and rapid method for the determination of eight food additives by RP-HPLC is described. They were saccharin, aspartame, benzoic acid, sorbic acid, vanillin, caffeine, carmine and sunset yellow. The experiments were carried on Shim-pack CLC-ODS (150 mm x 6.0 mm i.d.) with methanol-20 mmol/L NH4Ac (44:56, V/V; pH 7.0) as the eluent at a flow rate of 1.0 mL/min. The UV detection wavelength was fixed at 220 nm. The food samples, after precipitation of the impurities with Carrez reagent, were injected directly into the HPLC system. The average recoveries of all the eight additives were between 91.9%-108.5%, and the RSDs were lower than 4% (n = 5). The analysis of a single sample required only 8 min. This method has been successfully applied to the routine analysis of these additives in foods.

  2. Triethylborate as an electrolyte additive for high voltage layered lithium nickel cobalt manganese oxide cathode of lithium ion battery

    NASA Astrophysics Data System (ADS)

    Wang, Zaisheng; Xing, Lidan; Li, JianHui; Xu, Mengqing; Li, Weishan

    2016-03-01

    Triethylborate (TEB) is used as an electrolyte additive to improve the electrochemical performances of LiNi1/3Co1/3Mn1/3O2 (LNCM) upon cycling at 4.5 V vs. Li/Li+. Charge/discharge tests demonstrate that the cyclic stability of LNCM at room and elevated temperature can be improved effectively by TEB. With addition of 10 wt. % TEB into STD electrolyte (1.0 M LiPF6/EC:EMC:DEC), LNCM achieves a capacity retention of 99.8% after 150 cycles and 94.7% after 120 cycles at room and elevated temperature, respectively, comparing to that of 68.9% and 68.8% of STD electrolyte. In addition, 10 wt. % TEB also improves the rate capability of LNCM at room temperature. Physical and electrochemical characterizations from XRD, SEM, TEM, XPS, ICP-MS, LSV, CA, and EIS reveal that the preferential oxidative reaction of TEB generates a thin, uniform and low interfacial resistance film on the LNCM surface. This film not only suppresses the subsequent decomposition of STD electrolyte, but also prevents the dissolution of transition metal ions from LNCM, resulting in improved cyclic stability and rate capability of LNCM.

  3. Coffee cysteine proteinases and related inhibitors with high expression during grain maturation and germination

    PubMed Central

    2012-01-01

    Background Cysteine proteinases perform multiple functions in seeds, including participation in remodelling polypeptides and recycling amino acids during maturation and germination. Currently, few details exist concerning these genes and proteins in coffee. Furthermore, there is limited information on the cysteine proteinase inhibitors which influence the activities of these proteinases. Results Two cysteine proteinase (CP) and four cysteine proteinase inhibitor (CPI) gene sequences have been identified in coffee with significant expression during the maturation and germination of coffee grain. Detailed expression analysis of the cysteine proteinase genes CcCP1 and CcCP4 in Robusta using quantitative RT-PCR showed that these transcripts accumulate primarily during grain maturation and germination/post germination. The corresponding proteins were expressed in E. coli and purified, but only one, CcCP4, which has a KDDL/KDEL C-terminal sequence, was found to be active after a short acid treatment. QRT-PCR expression analysis of the four cysteine proteinase inhibitor genes in Robusta showed that CcCPI-1 is primarily expressed in developing and germinating grain and CcCPI-4 is very highly expressed during the late post germination period, as well as in mature, but not immature leaves. Transcripts corresponding to CcCPI-2 and CcCPI-3 were detected in most tissues examined at relatively similar, but generally low levels. Conclusions Several cysteine proteinase and cysteine proteinase inhibitor genes with strong, relatively specific expression during coffee grain maturation and germination are presented. The temporal expression of the CcCP1 gene suggests it is involved in modifying proteins during late grain maturation and germination. The expression pattern of CcCP4, and its close identity with KDEL containing CP proteins, implies this proteinase may play a role in protein and/or cell remodelling during late grain germination, and that it is likely to play a strong role

  4. Expressed sequence tag (EST) profiling in hyper saline shocked Dunaliella salina reveals high expression of protein synthetic apparatus components.

    PubMed

    Alkayal, Fadi; Albion, Rebecca L; Tillett, Richard L; Hathwaik, Leyla T; Lemos, Mark S; Cushman, John C

    2010-11-01

    The unicellular, halotolerant, green alga, Dunaliella salina (Chlorophyceae) has the unique ability to adapt and grow in a wide range of salt conditions from about 0.05 to 5.5M. To better understand the molecular basis of its salinity tolerance, a complementary DNA (cDNA) library was constructed from D. salina cells adapted to 2.5M NaCl, salt-shocked at 3.4M NaCl for 5h, and used to generate an expressed sequence tag (EST) database. ESTs were obtained for 2831 clones representing 1401 unique transcripts. Putative functions were assigned to 1901 (67.2%) ESTs after comparison with protein databases. An additional 154 (5.4%) ESTs had significant similarity to known sequences whose functions are unclear and 776 (27.4%) had no similarity to known sequences. For those D. salina ESTs for which functional assignments could be made, the largest functional categories included protein synthesis (35.7%), energy (photosynthesis) (21.4%), primary metabolism (13.8%) and protein fate (6.8%). Within the protein synthesis category, the vast majority of ESTs (80.3%) encoded ribosomal proteins representing about 95% of the approximately 82 subunits of the cytosolic ribosome indicating that D. salina invests substantial resources in the production and maintenance of protein synthesis. The increased mRNA expression upon salinity shock was verified for a small set of selected genes by real-time, quantitative reverse-transcription-polymerase chain reaction (qRT-PCR). This EST collection also provided important new insights into the genetic underpinnings for the biosynthesis and utilization of glycerol and other osmoprotectants, the carotenoid biosynthetic pathway, reactive oxygen-scavenging enzymes, and molecular chaperones (heat shock proteins) not described previously for D. salina. EST discovery also revealed the existence of RNA interference and signaling pathways associated with osmotic stress adaptation. The unknown ESTs described here provide a rich resource for the identification

  5. Expression of high frequency-losses in Mn-Zn ferrite

    SciTech Connect

    Fujiwara, T.; Tahara, R. )

    1993-11-01

    This paper gives a numerical expression for power losses at high frequencies (100 kHz to 2 MHz) in Mn-Zn ferrites, which are used as the core materials of transformers and inductors for switching power supplies. The power losses of some commercial ferrites were measured, and a new type of empirical formula was derived to express the power loss as a function of frequency and magnetic flux density. The calculated results agree well with measurements. The formula will prove useful in the design of compact and efficient magnetic components.

  6. High-yield production of canine parvovirus virus-like particles in a baculovirus expression system.

    PubMed

    Jin, Hongli; Xia, Xiaohong; Liu, Bing; Fu, Yu; Chen, Xianping; Wang, Huihui; Xia, Zhenqiang

    2016-03-01

    An optimized VP2 gene from the current prevalent CPV strain (new CPV-2a) in China was expressed in a baculovirus expression system. It was found that the VP2 proteins assembled into virus-like particles (VLPs) with antigenic properties similar to those of natural CPV and with an especially high hemagglutination (HA) titer (1:2(20)). Dogs intramuscularly or orally immunized with VLPs produced antibodies against CPV with >1:80 hemagglutination inhibition (HI) units for at least 3 months. The CPV VLPs could be considered for use as a vaccine against CPV or as a platform for research on chimeric VLP vaccines against other diseases.

  7. Simvastatin and Dipentyl Phthalate Lower Ex vivo Testicular Testosterone Production and Exhibit Additive Effects on Testicular Testosterone and Gene Expression Via Distinct Mechanistic Pathways in the Fetal Rat

    EPA Science Inventory

    Sex differentiation of the male reproductive tract in mammals is driven, in part, by fetal androgen production. In utero, some phthalate esters (PEs) alter fetal Leydig cell differentiation, reducing the expression of several genes associated with steroid synthesis/transport, and...

  8. The effect of nitrogen additions on oak foliage and herbivore communities at sites with high and low atmospheric pollution.

    PubMed

    Eatough Jones, Michele; Paine, Timothy D; Fenn, Mark E

    2008-02-01

    To evaluate plant and herbivore responses to nitrogen we conducted a fertilization study at a low and high pollution site in the mixed conifer forests surrounding Los Angeles, California. Contrary to expectations, discriminant function analysis of oak herbivore communities showed significant response to N fertilization when atmospheric deposition was high, but not when atmospheric deposition was low. We hypothesize that longer-term fertilization treatments are needed at the low pollution site before foliar N nutrition increases sufficiently to affect herbivore communities. At the high pollution site, fertilization was also associated with increased catkin production and higher densities of a byturid beetle that feeds on the catkins of oak. Leaf nitrogen and nitrate were significantly higher at the high pollution site compared to the low pollution site. Foliar nitrate concentrations were positively correlated with abundance of sucking insects, leafrollers and plutellids in all three years of the study.

  9. An Ultra-High Fluorescence Enhancement and High Throughput Assay for Revealing Expression and Internalization of Chemokine Receptor CXCR4.

    PubMed

    He, Hua; Wang, Xiaojuan; Cheng, Tiantian; Xia, Yongqing; Lao, Jun; Ge, Baosheng; Ren, Hao; Khan, Naseer Ullah; Huang, Fang

    2016-04-18

    Revealing chemokine receptor CXCR4 expression, distribution, and internalization levels in different cancers helps to evaluate cancer progression or prognosis and to set personalized treatment strategy. We here describe a sensitive and high-throughput immunoassay for determining CXCR4 expression and distribution in cancer cells. The assay is accessible to a wide range of users in an ordinary lab only by dip-coating poly(styrene-co-N-isopropylacrylamide) spheres on the glass substrate. The self- assembled spheres form three-dimensional photonic colloidal crystals which enhance the fluorescence of CF647 and Alexa Fluor 647 by a factor of up to 1000. CXCR4 in cells is detected by using the sandwich immunoassay, where the primary antibody recognizes CXCR4 and the secondary antibody is labeled with CF647. With the newly established assay, we quantified the total expression of CXCR4, its distribution on the cell membrane and cytoplasm, and revealed their internalization level upon SDF-1α activation in various cancer cells, even for those with extremely low expression level. PMID:26879206

  10. Constitutive high-level SOS1 expression and absence of HKT1;1 expression in the salt-accumulating halophyte Salicornia dolichostachya.

    PubMed

    Katschnig, D; Bliek, T; Rozema, J; Schat, H

    2015-05-01

    We investigated the effects of salinity on ion accumulation and expression of candidate salt tolerance genes in the highly tolerant salt accumulating halophyte Salicornia dolichostachya and the taxonomically related glycophytic Spinacia oleracea. S. dolichostachya, in comparison with S. oleracea, constitutively expressed SOS1 at a high level, but did not detectably express HKT1;1. These findings suggest that the constitutive high level of shoot salt accumulation in S. dolichostachya is accomplished through enhancement of SOS1-mediated Na(+) xylem loading, in combination with complete suppression of HKT1;1-mediated Na(+) retrieval from the xylem. Our findings demonstrate the importance of gene expression comparisons between highly tolerant halophytes and taxonomically related glycophytes to improve the understanding of mechanisms of Na(+) movement and salt tolerance in plants. PMID:25804817

  11. High levels of class III β-tubulin expression are associated with aggressive tumor features in breast cancer

    PubMed Central

    LEBOK, PATRICK; ÖZTÜRK, MELIKE; HEILENKÖTTER, UWE; JAENICKE, FRITZ; MÜLLER, VOLKMAR; PALUCHOWSKI, PETER; GEIST, STEFAN; WILKE, CHRISTIAN; BURANDT, EICKE; LEBEAU, ANNETTE; WILCZAK, WALDEMAR; KRECH, TILL; SIMON, RONALD; SAUTER, GUIDO; QUAAS, ALEXANDER

    2016-01-01

    Overexpression of class III β-tubulin (TUBB3), a factor that confers dynamic properties to microtubules, is a candidate biomarker for resistance to microtubule-targeting chemotherapeutics in breast and other types of solid cancer. Discrepant results from previous studies, with respect to the association of TUBB3 expression levels with breast cancer phenotype and patient prognosis, prompted the present study to investigate TUBB3 expression in a large cohort of breast cancer cases, with available clinical follow-up data. A preexisting breast cancer prognosis tissue microarray, containing a single 0.6 mm tissue core from each of 2,197 individual patients with breast cancer, was analyzed for TUBB3 expression by immunohistochemistry. The results of the present study revealed that TUBB3 expression was less frequent in lobular breast cancer cases (34%), compared with that of cancer cases of alternative histologies, including breast cancer of no special type (60%; P<0.0001). High TUBB3 positivity was associated with high tumor grade (P<0.0001), negativity for estrogen (P<0.0001) and progesterone receptors (P<0.004), as well as the presence of human epidermal growth factor 2 amplification (P<0.0001) and a triple-negative phenotype (P<0.0001). TUBB3 overexpression was additionally associated with reduced patient survival if all breast cancer cases of any histology were jointly analyzed (P=0.0088); however this link was not evident in the subset of breast cancer cases of no special type, or in a multivariate analysis including the established prognostic factors of tumor stage, grade and nodal stage. In conclusion, the present study demonstrated that TUBB3 overexpression was associated with adverse features of breast cancer, and that TUBB3 may possess a distinct role in lobular breast cancer cases, compared with alternative histological subtypes. The results of the present study do not support a clinically relevant role for TUBB3 as a prognostic marker in breast cancer. PMID

  12. Methods for efficient high-throughput screening of protein expression in recombinant Pichia pastoris strains.

    PubMed

    Camattari, Andrea; Weinhandl, Katrin; Gudiminchi, Rama K

    2014-01-01

    The methylotrophic yeast Pichia pastoris is becoming one of the favorite industrial workhorses for protein expression. Due to the widespread use of integration vectors, which generates significant clonal variability, screening methods allowing assaying hundreds of individual clones are of particular importance. Here we describe methods to detect and analyze protein expression, developed in a 96-well format for high-throughput screening of recombinant P. pastoris strains. The chapter covers essentially three common scenarios: (1) an enzymatic assay for proteins expressed in the cell cytoplasm, requiring cell lysis; (2) a whole-cell assay for a fungal cytochrome P450; and (3) a nonenzymatic assay for detection and quantification of tagged protein secreted into the supernatant. PMID:24744029

  13. Highly Expressed Genes within Hippocampal Sector CA1: Implications for the Physiology of Memory.

    PubMed

    Meyer, Michael A

    2014-04-22

    As the CA1 sector has been implicated to play a key role in memory formation, a dedicated search for highly expressed genes within this region was made from an on-line atlas of gene expression within the mouse brain (GENSAT). From a data base of 1013 genes, 16 were identified that had selective localization of gene expression within the CA1 region, and included Angpt2, ARHGEF6, CCK, Cntnap1, DRD3, EMP1, Epha2, Itm2b, Lrrtm2, Mdk, PNMT, Ppm1e, Ppp2r2d, RASGRP1, Slitrk5, and Sstr4. Of the 16 identified, the most selective and intense localization for both adult and post-natal day 7 was noted for ARHGEF6, which is known to be linked to non-syndromic mental retardation, and has also been localized to dendritic spines. Further research on the role played by ARHGEF6 in memory formation is strongly advocated. PMID:24987507

  14. Highly Expressed Genes within Hippocampal Sector CA1: Implications for the Physiology of Memory

    PubMed Central

    Meyer, Michael A.

    2014-01-01

    As the CA1 sector has been implicated to play a key role in memory formation, a dedicated search for highly expressed genes within this region was made from an on-line atlas of gene expression within the mouse brain (GENSAT). From a data base of 1013 genes, 16 were identified that had selective localization of gene expression within the CA1 region, and included Angpt2, ARHGEF6, CCK, Cntnap1, DRD3, EMP1, Epha2, Itm2b, Lrrtm2, Mdk, PNMT, Ppm1e, Ppp2r2d, RASGRP1, Slitrk5, and Sstr4. Of the 16 identified, the most selective and intense localization for both adult and post-natal day 7 was noted for ARHGEF6, which is known to be linked to non-syndromic mental retardation, and has also been localized to dendritic spines. Further research on the role played by ARHGEF6 in memory formation is strongly advocated PMID:24987507

  15. Evaluation of Vapor Pressure and Ultra-High Vacuum Tribological Properties of Ionic Liquids (2) Mixtures and Additives

    NASA Technical Reports Server (NTRS)

    Morales, Wilfredo; Koch, Victor R.; Street, Kenneth W., Jr.; Richard, Ryan M.

    2008-01-01

    Ionic liquids are salts, many of which are typically viscous fluids at room temperature. The fluids are characterized by negligible vapor pressures under ambient conditions. These properties have led us to study the effectiveness of ionic liquids containing both organic cations and anions for use as space lubricants. In the previous paper we have measured the vapor pressure and some tribological properties of two distinct ionic liquids under simulated space conditions. In this paper we will present vapor pressure measurements for two new ionic liquids and friction coefficient data for boundary lubrication conditions in a spiral orbit tribometer using stainless steel tribocouples. In addition we present the first tribological data on mixed ionic liquids and an ionic liquid additive. Post mortem infrared and Raman analysis of the balls and races indicates the major degradation pathway for these two organic ionic liquids is similar to those of other carbon based lubricants, i.e. deterioration of the organic structure into amorphous graphitic carbon. The coefficients of friction and lifetimes of these lubricants are comparable to or exceed these properties for several commonly used space oils.

  16. The effects of high-pressure treatments on Campylobacter jejuni in ground poultry products containing polyphosphate additives.

    PubMed

    Gunther, Nereus W; Sites, Joseph; Sommers, Christopher

    2015-09-01

    Marinades containing polyphosphates have been previously implicated in the enhanced survival of Campylobacter spp. in poultry product exudates. The enhanced Campylobacter survival has been attributed primarily to the ability of some polyphosphates to change the pH of the exudate to one more amenable to Campylobacter. In this study a ground poultry product contaminated with a 6 strain Campylobacter jejuni cocktail was utilized to determine if the efficiency of high-hydrostatic-pressure treatments was negatively impacted by the presence of commonly utilized polyphosphates. Two polyphosphates, hexametaphosphate and sodium tripolyphosphate, used at 2 concentrations, 0.25 and 0.5%, failed to demonstrate any significant negative effects on the efficiency of inactivation of C. jejuni by high-pressure treatment. However, storage at 4°C of the ground poultry samples containing C. jejuni after high-pressure treatment appeared to provide a synergistic effect on Campylobacter inactivation. High-pressure treatment in conjunction with 7 d of storage at 4°C resulted in a mean reduction in C. jejuni survival that was larger than the sum of the individual reductions caused by high pressure or 4°C storage when applied separately.

  17. High-level expression of a phage display-derived scFv in Pichia pastoris.

    PubMed

    Damasceno, Leonardo M; Lee, Frank; Ritter, Gerd; Old, Lloyd; Batt, Carl

    2009-01-01

    Numerous techniques are available for investigating protein-ligand interactions. The phage display technique is one such method routinely used to identify antibody-antigen interactions and has the benefit of being easily adaptable to high-throughput screening platforms. Once identified, antigen-binding domains on fragment antibodies or single-chain fragment antibodies (scFv) can be expressed and purified for further studies. In this chapter, we describe a method for high-level expression of a phage display-derived scFv in Pichia pastoris. The phage display-derived antibody A33scFv recognizes a cell surface glycoprotein (designated A33) expressed in colon cancer that serves as a target antigen for radioimmunoimaging and/or immunotherapy of human colon cancer. The expression and purification of A33scFv was optimized for the methylotrophic yeast P. pastoris. P. pastoris with a Mut(S) phenotype was selected to express A33scFv under regulation of the methanol-inducible AOX1 promoter. Here we describe a large-scale fed-batch fermentation process with an efficient online closed-loop methanol control for the production of the recombinant protein. Purification of A33scFv from clarified culture medium was done using a two-step chromatographic procedure using anion exchange and hydrophobic interaction chromatography, resulting in a final product with more than 90% purity. This chapter provides protocols that can be used as a base for process development of recombinant protein expression in P. pastoris and purification of these proteins for use in further functionality studies and in diagnostic and therapeutic applications.

  18. High cell density cultivation and high recombinant protein production of Escherichia coli strain expressing uricase.

    PubMed

    Nakagawa, S; Oda, H; Anazawa, H

    1995-12-01

    Uricase from Cellulomonas flavigena SK-4 is an industrially useful enzyme for commercial formulations of hair coloring. The uricase production by recombinant Escherichia coli strain with a high cell density cultivation technique was described. Of three kinds of media, synthetic media with the feeding of a high concentration of glucose solution were suitable for high cell density cultivation. As for feeding, both biomass concentration and uricase productivity were increased by about two (61.2 g dry cell weight (DCW)/liter) and three times (1037 U/ml broth), respectively, in 24 h by continuous supply. In the case of feeding by a DO-stat method, however, cell concentration was comparable to continuous glucose supply but uricase activity was reduced. By supplying pure oxygen to compensate for oxygen limitation during cultivation, the highest values of 77.4g DCW/liter and 1113 U/ml broth of the uricase activity were achieved with the total cultivation time of 15 h.

  19. Validation of the Amsterdam Dynamic Facial Expression Set – Bath Intensity Variations (ADFES-BIV): A Set of Videos Expressing Low, Intermediate, and High Intensity Emotions

    PubMed Central

    Wingenbach, Tanja S. H.

    2016-01-01

    Most of the existing sets of facial expressions of emotion contain static photographs. While increasing demand for stimuli with enhanced ecological validity in facial emotion recognition research has led to the development of video stimuli, these typically involve full-blown (apex) expressions. However, variations of intensity in emotional facial expressions occur in real life social interactions, with low intensity expressions of emotions frequently occurring. The current study therefore developed and validated a set of video stimuli portraying three levels of intensity of emotional expressions, from low to high intensity. The videos were adapted from the Amsterdam Dynamic Facial Expression Set (ADFES) and termed the Bath Intensity Variations (ADFES-BIV). A healthy sample of 92 people recruited from the University of Bath community (41 male, 51 female) completed a facial emotion recognition task including expressions of 6 basic emotions (anger, happiness, disgust, fear, surprise, sadness) and 3 complex emotions (contempt, embarrassment, pride) that were expressed at three different intensities of expression and neutral. Accuracy scores (raw and unbiased (Hu) hit rates) were calculated, as well as response times. Accuracy rates above chance level of responding were found for all emotion categories, producing an overall raw hit rate of 69% for the ADFES-BIV. The three intensity levels were validated as distinct categories, with higher accuracies and faster responses to high intensity expressions than intermediate intensity expressions, which had higher accuracies and faster responses than low intensity expressions. To further validate the intensities, a second study with standardised display times was conducted replicating this pattern. The ADFES-BIV has greater ecological validity than many other emotion stimulus sets and allows for versatile applications in emotion research. It can be retrieved free of charge for research purposes from the corresponding author

  20. Characterization of a Monoclonal Antibody Against CREPT, a Novel Protein Highly Expressed in Tumors

    PubMed Central

    Ren, Fangli; Wang, Ruoke; Zhang, Yanquan; Liu, Chunxiao; Wang, Yinyin; Hu, Jim; Zhang, Linqi

    2014-01-01

    CREPT (cell-cycle related and expression-elevated protein in tumor), a novel gene also called RPRD1B and C20ORF77, was recently identified to promote tumorigenesis through up-regulation of the expression of genes related to cell cycle. The previous study demonstrated that CREPT is highly expressed in a variety of tumors and enhances the expression of Cyclin D1 by promoting the formation of a chromatin loop. To study the correlation of CREPT expression with clinical factors in different tumors, we generated a monoclonal antibody (3E10) using purified recombinant human GST-CREPT protein as an antigen. In this study, we characterized the specificity of the monoclonal antibody and cloned the gene encoding the antibody for preparation of industrial production. Our results showed that the monoclonal antibody 3E10 was sensitive and specific to recognize human endogenous CREPT protein. We have mapped the epitope of the antibody and cloned the variable region sequence of the gene encoding the antibody. We confirmed that the cloned gene produced an equivalent antibody as that produced by the original hybridoma. This study provided a basis for large-scale production of the CREPT antibody, which will be useful for the study of the role of CREPT in different tumors. PMID:25545209

  1. Patterning Expression of Regenerative Growth Factors Using High Intensity Focused Ultrasound

    PubMed Central

    Wilson, Christopher G.; Martín-Saavedra, Francisco M.; Padilla, Frédéric; Fabiilli, Mario L.; Zhang, Man; Baez, Alexander M.; Bonkowski, Christopher J.; Kripfgans, Oliver D.; Voellmy, Richard; Vilaboa, Nuria; Fowlkes, J. Brian

    2014-01-01

    Temporal and spatial control of growth factor gradients is critical for tissue patterning and differentiation. Reinitiation of this developmental program is also required for regeneration of tissues during wound healing and tissue regeneration. Devising methods for reconstituting growth factor gradients remains a central challenge in regenerative medicine. In the current study we develop a novel gene therapy approach for temporal and spatial control of two important growth factors in bone regeneration, vascular endothelial growth factor, and bone morphogenetic protein 2, which involves application of high intensity focused ultrasound to cells engineered with a heat-activated- and ligand-inducible gene switch. Induction of transgene expression was tightly localized within cell-scaffold constructs to subvolumes of ∼30 mm3, and the amplitude and projected area of transgene expression was tuned by the intensity and duration of ultrasound exposure. Conditions for ultrasound-activated transgene expression resulted in minimal cytotoxicity and scaffold damage. Localized regions of growth factor expression also established gradients in signaling activity, suggesting that patterns of growth factor expression generated by this method will have utility in basic and applied studies on tissue development and regeneration. PMID:24460731

  2. High-Affinity PEGylated Polyacridine Peptide Polyplexes Mediate Potent In Vivo Gene Expression

    PubMed Central

    Kizzire, Koby; Khargharia, Sanjib; Rice, Kevin G.

    2012-01-01

    PEGylated polyacridine peptides bind to plasmid DNA with high affinity to form unique polyplexes that possess a long circulatory half-life and are hydrodynamically (HD)-stimulated to produce efficient gene expression in the liver of mice. We previously demonstrated that (Acr-Lys)6-Cys-PEG5kDa stabilizes a 1 μg pGL3 dose for up to 1 hr in the circulation, resulting in HD-stimulated (saline only) gene expression in the liver, equivalent in magnitude to direct-HD dosing of 1 μg of pGL3 (Fernandez C.A. et al. Gene Therapy 2011). In the present study we report that increasing the spacing of Acr with either 4 or 5 Lys residues, dramatically increases the stability of PEGylated polyacridine peptide polyplexes in the circulation allowing maximal HD-stimulated expression for up to 5 hrs post-DNA administration. Co-administration of a decoy dose of 9 μg of non-expressing DNA polyplex with 1 μg of pGL3 polyplex further extended the HD-stimulated expression to 9 hrs. This structure-activity relationship study defines the PEGylated polyacridine peptide requirements for maintaining fully transfection competent plasmid DNA in the circulation for 5 hrs and provides an understanding as to why polyplexes or lipoplexes prepared with PEI, chitosan or Lipofectamine are inactive within 5 min following i.v. dosing. PMID:22786534

  3. Clinical significance of CD81 expression by clonal plasma cells in high-risk smoldering and symptomatic multiple myeloma patients.

    PubMed

    Paiva, B; Gutiérrez, N-C; Chen, X; Vídriales, M-B; Montalbán, M-Á; Rosiñol, L; Oriol, A; Martínez-López, J; Mateos, M-V; López-Corral, L; Díaz-Rodríguez, E; Pérez, J-J; Fernández-Redondo, E; de Arriba, F; Palomera, L; Bengoechea, E; Terol, M-J; de Paz, R; Martin, A; Hernández, J; Orfao, A; Lahuerta, J-J; Bladé, J; Pandiella, A; Miguel, J-F San

    2012-08-01

    The presence of CD19 in myelomatous plasma cells (MM-PCs) correlates with adverse prognosis in multiple myeloma (MM). Although CD19 expression is upregulated by CD81, this marker has been poorly investigated and its prognostic value in MM remains unknown. We have analyzed CD81 expression by multiparameter flow cytometry in MM-PCs from 230 MM patients at diagnosis included in the Grupo Español de Mieloma (GEM)05>65 years trial as well as 56 high-risk smoldering MM (SMM). CD81 expression was detected in 45% (103/230) MM patients, and the detection of CD81(+) MM-PC was an independent prognostic factor for progression-free (hazard ratio=1.9; P=0.003) and overall survival (hazard ratio=2.0; P=0.02); this adverse impact was validated in an additional series of 325 transplant-candidate MM patients included in the GEM05 <65 years trial. Moreover, CD81(+) SMM (n=34/56, 57%) patients had a shorter time to progression to MM (P=0.02). Overall, our results show that CD81 may have a relevant role in MM pathogenesis and represent a novel adverse prognostic marker in myeloma. PMID:22333880

  4. High expression of N-myc (and STAT) interactor predicts poor prognosis and promotes tumor growth in human glioblastoma

    PubMed Central

    Yun, Dapeng; Zhao, Yingjie; Wang, Jingkun; Xu, Tao; Li, Xiaoying; Wang, Yuqi; Yuan, Li; Sun, Ruochuan; Song, Xiao; Huai, Cong; Hu, Lingna; Yang, Song; Min, Taishan; Chen, Juxiang; Chen, Hongyan; Lu, Daru

    2015-01-01

    Glioma is the most malignant brain tumor and glioblastoma (GBM) is the most aggressive type. The involvement of N-myc (and STAT) interactor (NMI) in tumorigenesis was sporadically reported but far from elucidation. This study aims to investigate roles of NMI in human glioma. Three independent cohorts, the Chinese tissue microarray (TMA) cohort (N = 209), the Repository for Molecular Brain Neoplasia Data (Rembrandt) cohort (N = 371) and The Cancer Genome Atlas (TCGA) cohort (N = 528 or 396) were employed. Transcriptional or protein levels of NMI expression were significantly increased according to tumor grade in all three cohorts. High expression of NMI predicted significantly unfavorable clinical outcome for GBM patients, which was further determined as an independent prognostic factor. Additionally, expression and prognostic value of NMI were associated with molecular features of GBM including PTEN deletion and EGFR amplification in TCGA cohort. Furthermore, overexpression or depletion of NMI revealed its regulation on G1/S progression and cell proliferation (both in vitro and in vivo), and this effect was partially dependent on STAT1, which interacted with and was regulated by NMI. These data demonstrate that NMI may serve as a novel prognostic biomarker and a potential therapeutic target for glioblastoma. PMID:25669971

  5. Mars Express High Resolution Stereo Camera spectrophotometric data: Characteristics and science analysis

    NASA Astrophysics Data System (ADS)

    McCord, T. B.; Adams, J. B.; Bellucci, G.; Combe, J.-P.; Gillespie, A. R.; Hansen, G.; Hoffmann, H.; Jaumann, R.; Neukum, G.; Pinet, P.; Poulet, F.; Stephan, K.

    2007-06-01

    The High Resolution Stereo Camera (HRSC) on the Mars Express (MEx) spacecraft in orbit about Mars is delivering images of the Mars surface and atmosphere from its high-inclination, elliptical orbit that are intended to cover most of the Mars surface by the end of the mission. These are acquired in four specific spectral passbands that sample the scene at different parts of the extended visual spectrum from near 0.4 μm to about 1μm and in five additional channels, four used for stereo and one nadir, that use the same fifth spectral passband. These channels view from different angles and can be used for photometric and spectral analysis. The data allow a variety of studies of the surface and atmosphere of Mars, but these require detailed understanding of the data characteristics, as there are several effects, including of the observation geometry and the atmosphere, that need to be treated. This article is a report to the science community on the current understanding of the HRSC color data after an initial analysis and also a demonstration of their capabilities for study of Mars, which, from this study, appear to be considerable. We first discuss the characteristics and calibration of the HRSC spectral data. The atmospheric influence on the photometric appearance of the surface materials is explored, and methods of compensating are considered. Reflectances are calculated for surface units by assuming the dark material is basalt and scaling the entire HRSC scene using a laboratory basalt spectrum for the selected dark material area; the results and the correction factors for different orbits are compared. We present several examples of what could become a large volume of future science results utilizing the HRSC spectrophotometric data in conjunction with photo-interpretation. For example, we find relatively few but very important distinct spectral components at the HRSC resolution, including what we interpret as iron oxide-rich material, unoxidized basalt, and

  6. Phosphazene additives

    DOEpatents

    Harrup, Mason K; Rollins, Harry W

    2013-11-26

    An additive comprising a phosphazene compound that has at least two reactive functional groups and at least one capping functional group bonded to phosphorus atoms of the phosphazene compound. One of the at least two reactive functional groups is configured to react with cellulose and the other of the at least two reactive functional groups is configured to react with a resin, such as an amine resin of a polycarboxylic acid resin. The at least one capping functional group is selected from the group consisting of a short chain ether group, an alkoxy group, or an aryloxy group. Also disclosed are an additive-resin admixture, a method of treating a wood product, and a wood product.

  7. Potlining Additives

    SciTech Connect

    Rudolf Keller

    2004-08-10

    In this project, a concept to improve the performance of aluminum production cells by introducing potlining additives was examined and tested. Boron oxide was added to cathode blocks, and titanium was dissolved in the metal pool; this resulted in the formation of titanium diboride and caused the molten aluminum to wet the carbonaceous cathode surface. Such wetting reportedly leads to operational improvements and extended cell life. In addition, boron oxide suppresses cyanide formation. This final report presents and discusses the results of this project. Substantial economic benefits for the practical implementation of the technology are projected, especially for modern cells with graphitized blocks. For example, with an energy savings of about 5% and an increase in pot life from 1500 to 2500 days, a cost savings of $ 0.023 per pound of aluminum produced is projected for a 200 kA pot.

  8. Comprehensive data base of high-level nuclear waste glasses: September 1987 status report: Volume 2, Additional appendices

    SciTech Connect

    Kindle, C.H.; Kreiter, M.R.

    1987-12-01

    The Materials Characterization Center (MCC) is assembling a comprehensive data base (CDB) of experimental data collected for high-level nuclear waste package components. The status of the CDB is summarized in Volume I of this report. Volume II contains appendices that present data from the data base and an evaluation of glass durability models applied to the data base.

  9. Addressing the Language and Literacy Needs of Aboriginal High School VET Students Who Speak SAE as an Additional Language

    ERIC Educational Resources Information Center

    Oliver, Rhonda; Grote, Ellen; Rochecouste, Judith; Exell, Mike

    2012-01-01

    Vocational Education and Training (VET) in high schools has had positive effects on the retention of Indigenous students, providing important pathways into further education and the workforce. However, low-level literacy (and numeracy) skills can make successful completion difficult, especially for students who speak Standard Australian English as…

  10. TBL1XR1 Is Highly Expressed in Gastric Cancer and Predicts Poor Prognosis

    PubMed Central

    He, Yuan; Liu, Ni; Zhang, Wenhui

    2016-01-01

    Objective. To investigate the expression of transducin- (β-) like 1 X-linked receptor 1 (TBL1XR1) in human gastric cancer (GC) and its correlation with prognostic and biologic significance. Methods. TBL1XR1 mRNA expression was analyzed in gastric cancer using a microarray dataset (GSE2701) from the Gene Expression Omnibus (GEO). Immunohistochemistry (IHC) analysis of TBL1XR1 was performed on GC tissue microarray (TMA) to assess its prognostic and biological significance in 334 patients of GC. Results. Analysis of GSE2701 showed that the mRNA levels of TBL1XR1 were significantly elevated in primary gastric tumor and lymph node tissues than normal gastric tissues (P < 0.05). The same results of TBL1XR1 protein level were observed by IHC staining in 334 GC tissues. 204 of 334 (60.1%) primary gastric cancer tissues showed high expression of TBL1XR1 protein. TBL1XR1 overexpression was significantly correlated with lymph node metastasis (P = 0.000) and advanced TNM stage (P = 0.001). Moreover, high levels of TBL1XR1 predicted worse overall survival (P = 0.015). Multivariate Cox regression analysis indicated that high expression of TBL1XR1 was an independent prognostic factor for poor overall survival (HR, 0.525; 95% confidence interval, 0.367–0.752; P = 0.005). Conclusion. This present study demonstrates that TBL1XR1 is overexpressed in gastric cancer and may be a potential predictor and therapeutic target for GC patients.

  11. Cyclooxygenase-2 (COX-2) expression in high-risk premalignant oral lesions.

    PubMed

    Sudbø, Jon; Ristimäki, Ari; Sondresen, Jan Erik; Kildal, Wanja; Boysen, Morten; Koppang, Hanna S; Reith, Albrecht; Risberg, Björn; Nesland, Jahn M; Bryne, Magne

    2003-07-01

    Emerging data indicate a link between genetic instability and up-regulation of cyclooxygenase-2 (COX-2). To see if individuals at high risk of oral cancer are candidates for treatment with selective COX-2 inhibitors (coxibs), levels of COX-2 expression in healthy, premalignant and cancerous oral mucosa were compared with the occurrence of DNA ploidy status as a genetic risk marker of oral cancer. COX-2 gene product was evaluated immunohistochemically in 30 healthy persons, in 22 patients with dysplastic lesions without previous or concomitant carcinomas, and in 29 patients with oral carcinomas. The immunohistochemical findings were verified by western blotting. COX-2 expression was correlated to DNA content as a genetic risk marker of oral cancer. COX-2 was up-regulated from healthy to premalignant to cancerous oral mucosa. Thus, COX-2 expression was found in 1 case of healthy oral mucosa (3%). All specimens from healthy mucosa had a normal DNA content. In patients with premalignancies. In 29 patients with oral carcinomas, cyclooxygenase-2 expression was observed in 26 (88%), and aneuploidy was observed in 25 cases (94%, P=0.04). Notably, of 22 patients with dysplastic lesions, COX-2 was exclusively expressed in a subgroup of nine patients (41%) identified to be at high risk of cancer by the aberrant DNA content of their lesions. Seven of these patients were followed for 5 years or more. An oral carcinoma developed in six of them (85%; P=0.02). These findings emphasize the need to determine whether coxibs can reduce the risk of oral cancer in patients with high-risk precancerous lesions. PMID:12747975

  12. TBL1XR1 Is Highly Expressed in Gastric Cancer and Predicts Poor Prognosis

    PubMed Central

    He, Yuan; Liu, Ni; Zhang, Wenhui

    2016-01-01

    Objective. To investigate the expression of transducin- (β-) like 1 X-linked receptor 1 (TBL1XR1) in human gastric cancer (GC) and its correlation with prognostic and biologic significance. Methods. TBL1XR1 mRNA expression was analyzed in gastric cancer using a microarray dataset (GSE2701) from the Gene Expression Omnibus (GEO). Immunohistochemistry (IHC) analysis of TBL1XR1 was performed on GC tissue microarray (TMA) to assess its prognostic and biological significance in 334 patients of GC. Results. Analysis of GSE2701 showed that the mRNA levels of TBL1XR1 were significantly elevated in primary gastric tumor and lymph node tissues than normal gastric tissues (P < 0.05). The same results of TBL1XR1 protein level were observed by IHC staining in 334 GC tissues. 204 of 334 (60.1%) primary gastric cancer tissues showed high expression of TBL1XR1 protein. TBL1XR1 overexpression was significantly correlated with lymph node metastasis (P = 0.000) and advanced TNM stage (P = 0.001). Moreover, high levels of TBL1XR1 predicted worse overall survival (P = 0.015). Multivariate Cox regression analysis indicated that high expression of TBL1XR1 was an independent prognostic factor for poor overall survival (HR, 0.525; 95% confidence interval, 0.367–0.752; P = 0.005). Conclusion. This present study demonstrates that TBL1XR1 is overexpressed in gastric cancer and may be a potential predictor and therapeutic target for GC patients. PMID:27672238

  13. High level of MT-MMP expression is associated with invasiveness of cervical cancer cells.

    PubMed

    Gilles, C; Polette, M; Piette, J; Munaut, C; Thompson, E W; Birembaut, P; Foidart, J M

    1996-01-17

    MMP-2 (gelatinase A) has been associated with the invasive potential of many cancer cells both in vitro and in vivo. It is now becoming clear that the activation of this enzyme might be a key step in tumor invasion. This activation process has been shown to be a membrane-associated pathway inducible by various agents such as collagen type I, concanavalin A or TGF-beta, but its physiological regulation is still largely unresolved. MT-MMP was recently discovered and described as a potential gelatinase-A activator. In the present study, we investigated the expression of MT-MMP (membrane-type metalloproteinase) in cervical cancer cells both in vitro and in vivo. Comparing several in vitro-transformed cervical cell lines, previously shown to display different invasive potentials, our results showed that the ability of cells to overexpress MT-MMP mRNA following ConA induction correlated with their ability to activate gelatinase A and with a highly invasive behavior. Moreover, using immunohistochemistry and in situ hybridization, we found a higher level of MT-MMP expression in invasive cervical carcinoma and lymph node metastases compared to its expression in non-invasive CIN III lesions. Our in vivo observations also clearly demonstrated a cooperation between stromal and tumor cells for the production of MT-MMP. Taken together, our results clearly correlated high level MT-MMP expression with invasiveness, and thus suggested that MT-MMP might play a crucial role in cervical tumor invasion.

  14. Addressing Shewanella oneidensis cytochromome : a first step towards high-throughput expression of cytochromes c.

    SciTech Connect

    Londer, Y. Y.; Giuliani, S.; Peppler, T.; Collart, F.; Biosciences Division

    2008-11-01

    Integrated studies that address proteins structure and function in the new era of systems biology and genomics often require the application of high-throughput approaches for parallel production of many different purified proteins from the same organism. Cytochromes c-electron transfer proteins carrying one or more hemes covalently bound to the polypeptide chain--are essential in most organisms. However, they are one of the most recalcitrant classes of proteins with respect to heterologous expression because post-translational incorporation of hemes is required for proper folding and stability. We have addressed this challenge by designing two families of vectors (total of 6 vectors) suitable for ligation-independent cloning and developing a pipeline for expression and solubility analysis of cytochromes c. This system has been validated by expression analysis of thirty genes from Shewanella oneidensis coding for cytochromes c or cytochromes c-type domains predicted to have 1-4 hemes. Out of 30 targets, 26 (87%) were obtained in soluble form in one or more vectors. This work establishes a methodology for high-throughput expression of this class of proteins and provides a clone resource for the microbiological and functional genomics research communities.

  15. Highly efficient integration and expression of piggyBac-derived cassettes in the honeybee (Apis mellifera)

    PubMed Central

    Schulte, Christina; Theilenberg, Eva; Müller-Borg, Marion; Gempe, Tanja; Beye, Martin

    2014-01-01

    Honeybees (Apis mellifera), which are important pollinators of plants, display remarkable individual behaviors that collectively contribute to the organization of a complex society. Advances in dissecting the complex processes of honeybee behavior have been limited in the recent past due to a lack of genetic manipulation tools. These tools are difficult to apply in honeybees because the unit of reproduction is the colony, and many interesting phenotypes are developmentally specified at later stages. Here, we report highly efficient integration and expression of piggyBac-derived cassettes in the honeybee. We demonstrate that 27 and 20% of queens stably transmitted two different expression cassettes to their offspring, which is a 6- to 30-fold increase in efficiency compared with those generally reported in other insect species. This high efficiency implies that an average beekeeping facility with a limited number of colonies can apply this tool. We demonstrated that the cassette stably and efficiently expressed marker genes in progeny under either an artificial or an endogenous promoter. This evidence of efficient expression encourages the use of this system to inhibit gene functions through RNAi in specific tissues and developmental stages by using various promoters. We also showed that the transgenic marker could be used to select transgenic offspring to be employed to facilitate the building of transgenic colonies via the haploid males. We present here the first to our knowledge genetic engineering tool that will efficiently allow for the systematic detection and better understanding of processes underlying the biology of honeybees. PMID:24821811

  16. High-resolution prediction of mouse brain connectivity using gene expression patterns.

    PubMed

    Fakhry, Ahmed; Ji, Shuiwang

    2015-02-01

    The brain is a multi-level system in which the high-level functions are generated by low-level genetic mechanisms. Thus, elucidating the relationship among multiple brain levels via correlative and predictive analytics is an important area in brain research. Currently, studies in multiple species have indicated that the spatiotemporal gene expression patterns are predictive of brain wiring. Specifically, results on the worm Caenorhabditis elegans have shown that the prediction of neuronal connectivity using gene expression signatures yielded statistically significant results. Recent studies on the mammalian brain produced similar results at the coarse regional level. In this study, we provide the first high-resolution, large-scale integrative analysis of the transcriptome and connectome in a single mammalian brain at a fine voxel level. By using the Allen Brain Atlas data, we predict voxel-level brain connectivity based on the gene expressions in the adult mouse brain. We employ regularized models to show that gene expression is predictive of connectivity at the voxel-level with an accuracy of 93%. We also identify a set of genes playing the most important role in connectivity prediction. We use only this small number of genes to predict the brain wiring with an accuracy over 80%. We discover that these important genes are enriched in neurons as compared to glia, and they perform connectivity-related functions. We perform several interesting correlative studies to further elucidate the transcriptome-connectome relationship.

  17. Selection of high expressing mammalian cells by surface display of reporters.

    PubMed

    DeMaria, Christine T

    2012-01-01

    A flow cytometry method using a nonfluorescent reporter protein was developed for rapid, early-stage identification of cells producing high levels of a recombinant protein of interest. A cell surface reporter protein is coexpressed with the protein of interest, and the reporter protein is detected using a fluorescently labeled antibody. The genes encoding the reporter protein and the protein of interest are linked by an IRES so that they are transcribed in the same mRNA but are translated independently. Since they each arise from a common mRNA, the reporter protein's expression level accurately predicts, on a per cell basis, the relative expression level of the protein of interest. This method provides an effective process for selecting cells that express high levels of recombinant proteins, with the benefits of rapid and accurate 96-well plate clone screening (that is both quantitative and qualitative) and elimination of unstable clones during subsequent scale up and culture. Furthermore, because this method does not rely on the availability of a detection reagent specific for the protein of interest that is expressed, it can be easily implemented into any cell line development process. PMID:21987245

  18. The Addition of Noncondensable Gases into RELAP5-3D for Analysis of High Temperature Gas-Cooled Reactors

    SciTech Connect

    C. B. Davis; C. H. Oh

    2003-08-01

    Oxygen, carbon dioxide, and carbon monoxide have been added to the RELAP5-3D computer code as noncondensable gases to support analysis of high temperature gas-cooled reactors. Models of these gases are required to simulate the effects of air ingress on graphite oxidation following a loss-of-coolant accident. Correlations were developed for specific internal energy, thermal conductivity, and viscosity for each gas at temperatures up to 3000 K. The existing model for internal energy (a quadratic function of temperature) was not sufficiently accurate at these high temperatures and was replaced by a more general, fourth-order polynomial. The maximum deviation between the correlations and the underlying data was 2.2% for the specific internal energy and 7% for the specific heat capacity at constant volume. The maximum deviation in the transport properties was 4% for oxygen and carbon monoxide and 12% for carbon dioxide.

  19. Improving high temperature creep resistance of reduced activation steels by addition of nitrogen and intermediate heat treatment

    NASA Astrophysics Data System (ADS)

    Liu, W. B.; Zhang, C.; Xia, Z. X.; Yang, Z. G.

    2014-12-01

    In the present study, we report an enhanced high-temperature creep resistance in reduced activation ferrite/martensite (RAFM) steels, by introducing nitrogen (0.035 wt%, M3 steel) and employing a novel intermediate heat treatment I-Q-T (intermediate treatment, quenching and tempering). In comparison with all the control groups, the uniaxial tests of the I-Q-T treated M3 steel showed significant increase in rupture time and decrease in elongation. The microstructures of the samples were further characterized to elucidate the origin of the enhanced creep resistance. It is found that, by introducing nitrogen, the primary TaC particles were refined; by employing the I-Q-T heat treatment, the dispersed fine secondary MX precipitates, as well as the lath subgrains containing high-density dislocations, were increased: all are responsible for the improved creep resistance.

  20. Improvement of corrosion resistance of high-velocity oxyfuel-sprayed stainless steel coatings by addition of molybdenum

    NASA Astrophysics Data System (ADS)

    Kawakita, Jin; Kuroda, Seiji; Fukushima, Takeshi; Kodama, Toshiaki

    2005-06-01

    To improve the marine corrosion resistance of stainless steel coatings fabricated by high-velocity oxyfuel (HVOF) spraying with a gas shroud attachment, the molybdenum (Mo) content of stainless steel was increased to form coatings with a chemical composition of Fe balance-18mass%Cr-22mass%Ni-2˜8mass%Mo. These coatings were highly dense, with <0.1 vol.% in porosity, and less oxidized, with 0.5 mass% in oxygen content at most. The corrosion mechanism and resistance of the coatings were investigated by electrochemical measurement, chemical analysis, and statistical processing. The general corrosion resistance of the coatings in 0.5 mol/dm3 sulfuric acid was improved with increases in Mo content, and the corrosion rate could be decreased to 8.8 × 10-2 mg/cm2 per hour (˜1 mm/year) at 8 mass% Mo. The pitting corrosion resistance of the coatings in artificial seawater was improved with increases in Mo content and was superior to that of the 316L stainless steel coating. The crevice corrosion resistance of the coatings in artificial seawater was improved and the number of rust spots at 4 mass% Mo was decreased to 38% of that for the 316L coating. Accordingly, Mo is highly effective in improving the corrosion resistance of the stainless steel coatings by HVOF spraying.

  1. High expression of RELM-α correlates with poor prognosis and promotes angiogenesis in gastric cancer.

    PubMed

    Chen, Ping; Zhao, Deshou; Wang, Weiyi; Zhang, Yongping; Yuan, Yaozong; Wang, Lifu; Wu, Yunlin

    2015-07-01

    Accumulating evidence indicates that resistin-like molecule-α (RELM-α) is involved in angiogenesis, while the clinical significance and the exact role of RELM-α in gastric cancer remain obscure. The aim of the present study was to evaluate the clinical significance of RELM-α in gastric cancer, and to investigate its effective mechanisms in order to identify a potential therapeutic target. The expression levels of RELM-α in 92 gastric cancer and adjacent normal tissues were investigated and the relationship between RELM-α expression and the clinicopathological characteristics was explored. To investigate the potential role of RELM-α in gastric cancer cell biological behavior, the cell proliferation, migration and invasion assays were conducted using two gastric cancer cell lines (SGC7901 and MKN45). We also assessed whether RELM-α gene silencing modulates angiogenesis using small interference RNA in cancer cell lines, and investigated its effect on nuclear factor (NF)-κB activation and vascular endothelial growth factor (VEGF) and MMP-9 expression. Contrasting sharply with the strong RELM-α-positive tumors, adjacent normal tissues and cell lines exhibited negative or weakly positive expression (P<0.01). High expression level of RELM-α was associated with advanced stage and tumor size (P<0.01). The silencing of RELM-α expression by Ad5/F35-siRNA treatment significantly inhibited cell migratory and invasive ability in SGC7901 and MKN45 gastric cancer cells compared with the control and Ad5/F35 vector-transfected cell lines (P<0.01). However, the silencing of RELM-α expression also significantly blocked NF-κB activation and attenuated VEGF and MMP-9 expression. The data demonstrated that RELM-α is a promising novel biomarker of angiogenesis in patients with gastric cancer. The study identified that the silencing of RELM-α expression may regulate the proliferation, invasion and migration of gastric cancer cells by targeting VEGF/MMP-9, and the mechanism

  2. High expression of RELM-α correlates with poor prognosis and promotes angiogenesis in gastric cancer.

    PubMed

    Chen, Ping; Zhao, Deshou; Wang, Weiyi; Zhang, Yongping; Yuan, Yaozong; Wang, Lifu; Wu, Yunlin

    2015-07-01

    Accumulating evidence indicates that resistin-like molecule-α (RELM-α) is involved in angiogenesis, while the clinical significance and the exact role of RELM-α in gastric cancer remain obscure. The aim of the present study was to evaluate the clinical significance of RELM-α in gastric cancer, and to investigate its effective mechanisms in order to identify a potential therapeutic target. The expression levels of RELM-α in 92 gastric cancer and adjacent normal tissues were investigated and the relationship between RELM-α expression and the clinicopathological characteristics was explored. To investigate the potential role of RELM-α in gastric cancer cell biological behavior, the cell proliferation, migration and invasion assays were conducted using two gastric cancer cell lines (SGC7901 and MKN45). We also assessed whether RELM-α gene silencing modulates angiogenesis using small interference RNA in cancer cell lines, and investigated its effect on nuclear factor (NF)-κB activation and vascular endothelial growth factor (VEGF) and MMP-9 expression. Contrasting sharply with the strong RELM-α-positive tumors, adjacent normal tissues and cell lines exhibited negative or weakly positive expression (P<0.01). High expression level of RELM-α was associated with advanced stage and tumor size (P<0.01). The silencing of RELM-α expression by Ad5/F35-siRNA treatment significantly inhibited cell migratory and invasive ability in SGC7901 and MKN45 gastric cancer cells compared with the control and Ad5/F35 vector-transfected cell lines (P<0.01). However, the silencing of RELM-α expression also significantly blocked NF-κB activation and attenuated VEGF and MMP-9 expression. The data demonstrated that RELM-α is a promising novel biomarker of angiogenesis in patients with gastric cancer. The study identified that the silencing of RELM-α expression may regulate the proliferation, invasion and migration of gastric cancer cells by targeting VEGF/MMP-9, and the mechanism

  3. High-grade fimbrial-ovarian carcinomas are unified by altered p53, PTEN and PAX2 expression.

    PubMed

    Roh, Michael H; Yassin, Yosuf; Miron, Alexander; Mehra, Karishma K; Mehrad, Mitra; Monte, Nicolas M; Mutter, George L; Nucci, Marisa R; Ning, Geng; Mckeon, Frank D; Hirsch, Michelle S; Wa, Xian; Crum, Christopher P

    2010-10-01

    High-grade endometrioid and serous carcinomas of the ovary and fallopian tube are responsible for the majority of cancer deaths and comprise a spectrum that includes early or localized (tubal intraepithelial carcinoma) and advanced (invasive or metastatic) disease. We subdivided a series of these tumors into three groups, (1) classic serous, (2) mixed serous and endometrioid and (3) endometrioid carcinomas and determined: (1) the frequencies of coexisting tubal intraepithelial carcinoma, (2) frequency of a dominant ovarian mass suggesting an ovarian origin and (3) immuno-localization of WT-1, p53, PTEN, PAX2 and p16(ink4). All tumors were analyzed for p53 mutations. Thirty six, 25 and 8% of groups 1-3 were associated with tubal intraepithelial carcinoma (P=0.09) and 34, 45 and 62% predominated in one ovary (P=0.028), respectively. Differences in frequencies of diffuse p53 immunostaining (85-93%), WT-1 (70-98%) and p16(ink4) positivity (69-75%) were not significant for all groups. Greater than 95% reduction in PAX2 and PTEN occurred in 67-75 and 5-12%, respectively; however, PAX2 and PTEN staining intensity, when present, was often heterogeneous, highlighting different tumor populations. PAX2 and PTEN expression were markedly reduced or absent in 12 of 12 and 4 of 12 tubal intraepithelial carcinomas. In summary, high-grade müllerian carcinomas share identical frequencies of altered or reduced expression of p53, PTEN and PAX2, all of which can be appreciated in tubal intraepithelial carcinomas. Because only a subset of these tumors appears to arise in the fallopian tube, attention to expression of these biomarkers in the ovary and other müllerian sites might facilitate the identification of other carcinogenic pathways. PAX2 and PTEN, in addition to p53 and p16(ink4), comprise a potentially important gene combination in high-grade pelvic carcinogenesis.

  4. [Benign salivary gland-type tumors of the bronchus: expression of high molecular weight cytokeratins].

    PubMed

    Méjean-Lebreton, Frédérique; Barnoud, Raphaëlle; de la Roche, Eric; Devouassoux-Shisheboran, Mojgan

    2006-02-01

    Primary lung tumors showing features of salivary gland-type neoplasms are extremely rare, and their immunohistochemical profile has been seldom studied. We report two cases of bronchial pleomorphic and mucous gland adenomas and study the expression of markers such as TTF-1 and high molecular weight keratins in these tumors. Both tumors were endobronchial. The pleomorphic adenoma also had a well-circumscribed parenchymal component, with a biphasic morphology composed of epithelial and myoepithelial cells in a background of myxoid and hyaline stroma. The mucous gland adenoma displayed papillary and dilated glandular structures. In both cases, epithelial cells showed strong and diffuse cytoplasmic staining with high molecular weight cytokeratins (cytokeratin 5/6 and keratin 903), and lacked TTF-1 expression. This immunoprofile provides useful clues for the histogenesis of pulmonary benign salivary gland-type adenomas and helps in distinguishing them from primary adenocarcinomas in small biopsy specimens.

  5. Impact of high fat diet on long non-coding RNAs and messenger RNAs expression in the aortas of ApoE(−/−) mice

    PubMed Central

    Bao, Mei-hua; Luo, Huai-qing; Chen, Li-hua; Tang, Liang; Ma, Kui-fen; Xiang, Ju; Dong, Li-ping; Zeng, Jie; Li, Guang-yi; Li, Jian-ming

    2016-01-01

    Atherosclerosis is a chronic multifactorial inflammatory disease with high prevalence worldwide, and has become the leading cause of death. The present study was designed to investigate the impact of high-fat diet on ApoE(−/−) mice exhibiting atherosclerosis by detecting the genome-wide expression profile of lncRNAs and mRNAs. A total of 354 differentially expressed lncRNAs were identified (≥2.0 folds). Simultaneously, 357 differentially expressed mRNAs from the same chip were found. The expression differences of lncRNAs and mRNAs were consistent in both qPCR and microarray detection. Annotation results of the mRNAs which correlated with lncRNAs showed that the commonly related pathways were metabolism and inflammation. Hypergeometric distribution analysis indicated that the differentially expressed lncRNAs had been mostly regulated by transcription factors (TFs) such as Myod1, Rxra, Pparg, Tcf3, etc. Additional lncRNA-target-TFs network analysis was conducted for the top 20 differentially expressed lncRNAs. The results indicated Hnf4a, Ppara, Vdr, and Runx3 as the TFs most likely to regulate the production of these lncRNAs, and might play roles in inflammatory and metabolic processes in atherosclerosis. In a nutshell, the present study identified a panel of dysregulated lncRNAs and mRNAs that may be potential biomarkers or drug targets relevant to the high-fat diet related atherogenesis. PMID:27698357

  6. A thieno[3,2-b][1]benzothiophene isoindigo building block for additive- and annealing-free high-performance polymer solar cells.

    PubMed

    Yue, Wan; Ashraf, Raja Shahid; Nielsen, Christian B; Collado-Fregoso, Elisa; Niazi, Muhammad R; Yousaf, Syeda Amber; Kirkus, Mindaugas; Chen, Hung-Yang; Amassian, Aram; Durrant, James R; McCulloch, Iain

    2015-08-26

    A novel photoactive polymer with two different molecular weights is reported, based on a new building block: thieno[3,2-b][1]benzothiophene isoindigo. Due to the improved crystallinity, optimal blend morphology, and higher charge mobility, solar-cell devices of the high-molecular-weight polymer exhibit a superior performance, affording efficiencies of 9.1% without the need for additives, annealing, or additional extraction layers during device fabrication.

  7. The Effect of Processing Additives on Energetic Disorder in Highly Efficient Organic Photovoltaics: A Case Study on PBDTTT-C-T:PC71 BM.

    PubMed

    Gao, Feng; Himmelberger, Scott; Andersson, Mattias; Hanifi, David; Xia, Yuxin; Zhang, Shaoqing; Wang, Jianpu; Hou, Jianhui; Salleo, Alberto; Inganäs, Olle

    2015-07-01

    Energetic disorder, an important parameter affecting the performance of organic photovoltaics, is significantly decreased upon the addition of processing additives in a highly efficient benzodithiophene-based copolymer blend (PBDTTT-C-T:PC71 BM). Wide-angle and small-angle X-ray scattering measurements suggest that the origin of this reduced energetic disorder is due to increased aggregation and a larger average fullerene domain size together with purer phases.

  8. Rapid High-Level Production of Functional HIV Broadly Neutralizing Monoclonal Antibodies in Transient Plant Expression Systems

    PubMed Central

    Rosenberg, Yvonne; Sack, Markus; Montefiori, David; Forthal, Donald; Mao, Lingjun; -Abanto, Segundo Hernandez; Urban, Lori; Landucci, Gary; Fischer, Rainer; Jiang, Xiaoming

    2013-01-01

    Passive immunotherapy using anti-HIV broadly neutralizing monoclonal antibodies (mAbs) has shown promise as an HIV treatment, reducing mother-to-child-transmission (MTCT) of simian/human immunodeficiency virus (SHIV) in non-human primates and decreasing viral rebound in patients who ceased receiving anti-viral drugs. In addition, a cocktail of potent mAbs may be useful as mucosal microbicides and provide an effective therapy for post-exposure prophylaxis. However, even highly neutralizing HIV mAbs used today may lose their effectiveness if resistance occurs, requiring the rapid production of new or engineered mAbs on an ongoing basis in order to counteract the viral resistance or the spread of a certain HIV-1 clade in a particular region or patient. Plant-based expression systems are fast, inexpensive and scalable and are becoming increasingly popular for the production of proteins and monoclonal antibodies. In the present study, Agrobacterium-mediated transient transfection of plants, utilizing two species of Nicotiana, have been tested to rapidly produce high levels of an HIV 89.6PΔ140env and several well-studied anti-HIV neutralizing monoclonal antibodies (b12, 2G12, 2F5, 4E10, m43, VRC01) or a single chain antibody construct (m9), for evaluation in cell-based viral inhibition assays. The protein-A purified plant-derived antibodies were intact, efficiently bound HIV envelope, and were equivalent to, or in one case better than, their counterparts produced in mammalian CHO or HEK-293 cells in both neutralization and antibody dependent viral inhibition assays. These data indicate that transient plant-based transient expression systems are very adaptable and could rapidly generate high levels of newly identified functional recombinant HIV neutralizing antibodies when required. In addition, they warrant detailed cost-benefit analysis of prolonged incubation in plants to further increase mAb production. PMID:23533588

  9. Werner Complexes with ω-Dimethylaminoalkyl Substituted Ethylenediamine Ligands: Bifunctional Hydrogen-Bond-Donor Catalysts for Highly Enantioselective Michael Additions.

    PubMed

    Ghosh, Subrata K; Ganzmann, Carola; Bhuvanesh, Nattamai; Gladysz, John A

    2016-03-18

    The racemic carbonate complex [Co(en)2 O2 CO](+) Cl(-) (en=1,2-ethylenediamine) and (S)-[H3 NCH((CH2 )n NHMe2 )CH2 NH3 ](3+) 3 Cl(-) (n=1-4) react (water, charcoal, 100 °C) to give [Co(en)2 ((S)-H2 NCH((CH2 )n NHMe2 )CH2 NH2 )](4+) 4 Cl(-) (3 a-d H(4+) 4 Cl(-) ) as a mixture of Λ/Δ diastereomers that separate on chiral-phase Sephadex columns. These are treated with NaOH/Na(+) BArf (-) (BArf =B(3,5-C6 H3 (CF3 )2 )4 ) to give lipophilic Λ- and Δ-3 a-d(3+) 3 BArf (-) , which are screened as catalysts (10 mol %) for additions of dialkyl malonates to nitroalkenes. Optimal results are obtained with Λ-3 c(3+) 3 BArf (-) (CH2 Cl2 , -35 °C; 98-82 % yields and 99-93 % ee for six β-arylnitroethenes). The monofunctional catalysts Λ- and Δ-[Co(en)3 ](3+) 3 BArf (-) give enantioselectivities of <10 % ee with equal loadings of Et3 N. The crystal structure of Δ-3 a H(4+) 4 Cl(-) provides a starting point for speculation regarding transition-state assemblies. PMID:26918320

  10. The effect of alloying additions on the high temperature deformation characteristics of Ti-48Al (at%) alloys

    SciTech Connect

    Sabinash, C.M.; Sastry, S.M.L.; Jerina, K.L.

    1995-05-01

    Addition of ternary and quaternary alloying elements at levels between one and three atomic percent increase the room temperature ductility of Ti-48Al (at%) based titanium aluminides. In this investigation the deformation characteristics of Ti-48Al-2X (X = Cr, Mn) alloys at temperatures of 1,200 C to 1,300 C and strain rates of 10{sup {minus}3}s{sup {minus}1} to 10{sup {minus}1}s{sup {minus}1} were studied and compared with the deformation characteristics of Ti-48Al using the same conditions. Three titanium aluminide alloys with target compositions of Ti-48Al, Ti-48Al-2Cr, and Ti-48Al-2Mn (at%) were induction skull melted and cast into 69.85 mm. diameter ingots. The cast ingots were hot isostatically pressed (HIPed) at 103.5 MPa, 1,177 C for 4 hours to close casting porosity and homogenize the alloy microstructure and composition. Cylindrical test specimens 10 mm dia by 15 mm height were excised by EDM from the ingot and were subjected to isothermal compression testing in air at 1,200--1,300 C and the effective strain rates of 10{sup {minus}3}--10{sup {minus}1}s{sup {minus}1}.

  11. Synthesis of Highly Active Mg-BASED Hydrides Using Hydriding Combustion Synthesis and NbF5 Additives

    NASA Astrophysics Data System (ADS)

    Chourashiya, M. G.; Park, C. N.; Park, C. J.

    2012-09-01

    Superiority of the hydriding combustion (HC) technique over conventional metallurgical approach to the synthesis of cost-effective Mg based hydrides, which show promise as hydrogen storage materials, is well known. In the present research, we report further improvements in HC prepared Mg-based materials, achieved by optimizing the preparative parameters of HC and by catalytic addition. Mg90-Ni60-C40 composites prepared using optimized processing parameters were ball-milled with NbF5 (10 h) and characterized for their micro-structural and hydriding properties. The ball-milled/catalyzed powder showed decreased crystallinity with CNTs on its surfaces. Surface area of the ball-milled powder decreased to almost half of the as-HC powder, while TG analysis revealed a four-fold decrease in the desorption temperature of the milled powder compared to that of the as-HC prepared powder. Activated samples achieved the maximum absorption/desorption limits (5.3 wt.%) at as low as 100°C, underlining the possibility of the use of these materials in portable hydrogen storage devices.

  12. Differential miRNA expression in inherently high- and low-active inbred mice

    PubMed Central

    Dawes, Michelle; Kochan, Kelli J; Riggs, Penny K; Timothy Lightfoot, J

    2015-01-01

    Despite established health benefits of regular exercise, the majority of Americans do not meet the recommended levels of physical activity. While it is known that voluntary activity levels are largely heritable, the genetic mechanisms that regulate activity are not well understood. MicroRNAs (miRNAs) are small non-coding RNAs that inhibit transcription by binding to a target gene, inhibiting protein production. The purpose of this study was to investigate differential miRNA expression between inherently high- (C57L/J) and low- (C3H/HeJ) active inbred mice in soleus, extensor digitorum longus (EDL), and nucleus accumbens tissues. Expression was initially determined by miRNA microarray analysis, and selected miRNAs were validated by qRT-PCR. Expression of 13 miRNAs varied between strains in the nucleus accumbens, 20 in soleus, and eight in EDL, by microarray analysis. Two miRNAs were validated by qRT-PCR in the nucleus accumbens; miR-466 was downregulated (∼4 fold; P < 0.0004), and miR-342-5p was upregulated (∼115 fold; P < 0.0001) in high-active mice. MiR-466 was downregulated (∼5 fold; P < 0.0001) in the soleus of high-active mice as well. Interestingly, miR-466 is one of several miRNA families with sequence located in intron 10 of Sfmbt2; miRNAs at this locus are thought to drive imprinting of this gene. “Pathways in cancer” and “TGFβ signaling” were the most significant pathways of putative target genes in both the soleus and nucleus accumbens. Our results are the first to consider differential miRNA expression between high- and low-active mice, and suggest that miRNAs may play a role in regulation of physical activity. PMID:26229004

  13. Functional expression of the high affinity receptor for IgE (FcepsilonRI) in human platelets and its' intracellular expression in human megakaryocytes.

    PubMed

    Hasegawa, S; Pawankar, R; Suzuki, K; Nakahata, T; Furukawa, S; Okumura, K; Ra, C

    1999-04-15

    The high affinity IgE receptor (FcepsilonRI) expressed on the cell surface of mast cells and basophils is the key molecule in triggering the IgE-mediated allergic reaction. Recently, it was elucidated that the FcepsilonRI is expressed on a variety of other cells like Langerhans cells, monocytes, and eosinophils, and the functional importance of the FcepsilonRI expression in Langerhans cells was also shown. Some studies suggest that human platelets may play important roles in allergic inflammation through the cell-surface expression of the FcepsilonRII and FcgammaRII. Here, we report that human platelets and megakaryocytes constitutively express the messenger RNA and protein for the FcepsilonRI. Although the FcepsilonRI is expressed on the cell surface of human platelets, it is only detected in the cytoplasm of human megakaryocytes. We also confirmed that human platelets express the genes for the alpha, beta, and gamma chains of the FcepsilonRI without any defined mutations. Furthermore, stimulation of human platelets via the FcepsilonRI induced the release of serotonin and RANTES (Regulated on Activation, Normal T Expressed, and presumably Secreted). Taken together, these results suggest a novel and important role for human platelets in perpetuating allergic inflammation through the expression of and activation via the FcepsilonRI.

  14. High expression of C-C chemokine receptor 2 associates with poor overall survival in gastric cancer patients after surgical resection

    PubMed Central

    Lin, Chao; Cao, Yifan; Zhang, Weijuan; Shen, Zhenbin; Xu, Jiejie

    2016-01-01

    Background Being a critical chemokine receptor in chemoattracting myeloid cells into tumor tissues, C-C chemokine receptor 2 (CCR2) has been detected in many malignant tumors. This study aims to evaluate the pro