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Sample records for fusion protein designed

  1. Rational design of an EGF-IL18 fusion protein: Implication for developing tumor therapeutics

    International Nuclear Information System (INIS)

    Lu Jianxin; Peng Ying; Meng Zhefeng; Jin Liqin; Lu Yongsui; Guan Minxin

    2005-01-01

    Interleukin-18 (IL-18) is a proinflammatory cytokine. This protein has a role in regulating immune responses and exhibits significant anti-tumor activities. Epidermal growth factor (EGF) is an important growth factor that plays a central role in the regulation of cell cycle and differentiation. It was proposed that a targeted delivery of IL-18 by generation of IL-18-EGF fusion protein might decrease adverse effects and result in enhancing cytotoxic and antitumor activities. In the present study, a fusion protein, consisting of EGFR binding domain fused to human IL-18 mature peptide via a linker peptide of (Gly 4 Ser) 3, was constructed and expressed in the insect cell line Sf9 using Bac-to-Bac baculovirus expression system. We showed that the purified recombinant fusion protein induced similar levels of IFN-γ to that of native IL-18 protein in human PBMC in the presence of ConA. Furthermore, EGF receptor competitive test in human epithelial cancer A431 cell line showed that EGF-IL18 fusion protein can specifically bind with EGFR by competing with native EGF protein. These suggest that this rationally designed protein can be further developed as novel tumor therapeutics

  2. Rational design of highly potent HIV-1 fusion inhibitory proteins: Implication for developing antiviral therapeutics

    International Nuclear Information System (INIS)

    Ni Ling; Gao, George F.; Tien Po

    2005-01-01

    Recombinant protein containing one heptad-repeat 1 (HR1) segment and one HR2 segment of the HIV-1 gp41 (HR1-HR2) has been shown to fold into thermally stable six-helix bundle, representing the fusogenic core of gp41. In this study, we have used the fusogenic core as a scaffold to design HIV-1 fusion inhibitory proteins by linking another HR1 to the C terminus of HR1-HR2 (HR121) or additional HR2 to the N terminus of HR1-HR2 (HR212). Both recombinant proteins could be abundantly and solubly expressed and easily purified, exhibiting high stability and potent inhibitory activity on HIV-1 fusion with IC 50 values of 16.2 ± 2.8 and 2.8 ± 0.63 nM, respectively. These suggest that these rationally designed proteins can be further developed as novel anti-HIV-1 therapeutics

  3. Fusion reactor design studies

    International Nuclear Information System (INIS)

    Emmert, G.A.; Kulcinski, G.L.; Santarius, J.F.

    1990-01-01

    This report discusses the following topics on the ARIES tokamak: systems; plasma power balance; impurity control and fusion ash removal; fusion product ripple loss; energy conversion; reactor fueling; first wall design; shield design; reactor safety; and fuel cost and resources

  4. Biochemistry and biophysics of HIV-1 gp41 - membrane interactions and implications for HIV-1 envelope protein mediated viral-cell fusion and fusion inhibitor design.

    Science.gov (United States)

    Cai, Lifeng; Gochin, Miriam; Liu, Keliang

    2011-12-01

    Human immunodeficiency virus type 1 (HIV-1), the pathogen of acquired immunodeficiency syndrome (AIDS), causes ~2 millions death every year and still defies an effective vaccine. HIV-1 infects host cells through envelope protein - mediated virus-cell fusion. The transmembrane subunit of envelope protein, gp41, is the molecular machinery which facilitates fusion. Its ectodomain contains several distinguishing functional domains, fusion peptide (FP), Nterminal heptad repeat (NHR), C-terminal heptad repeat (CHR) and membrane proximal extracellular region (MPER). During the fusion process, FP inserts into the host cell membrane, and an extended gp41 prehairpin conformation bridges the viral and cell membranes through MPER and FP respectively. Subsequent conformational change of the unstable prehairpin results in a coiled-coil 6-helix bundle (6HB) structure formed between NHR and CHR. The energetics of 6HB formation drives membrane apposition and fusion. Drugs targeting gp41 functional domains to prevent 6HB formation inhibit HIV-1 infection. T20 (enfuvirtide, Fuzeon) was approved by the US FDA in 2003 as the first fusion inhibitor. It is a 36-residue peptide from the gp41 CHR, and it inhibits 6HB formation by targeting NHR and lipids. Development of new fusion inhibitors, especially small molecule drugs, is encouraged to overcome the shortcomings of T20 as a peptide drug. Hydrophobic characteristics and membrane association are critical for gp41 function and mechanism of action. Research in gp41-membrane interactions, using peptides corresponding to specific functional domains, or constructs including several interactive domains, are reviewed here to get a better understanding of gp41 mediated virus-cell fusion that can inform or guide the design of new HIV-1 fusion inhibitors.

  5. Design of Recombinant Stem Cell Factor macrophage Colony Stimulating Factor Fusion Proteins and their Biological Activity In Vitro

    Science.gov (United States)

    Chen, Tao; Yang, Jie; Wang, Yuelang; Zhan, Chenyang; Zang, Yuhui; Qin, Junchuan

    2005-05-01

    Stem cell factor (SCF) and macrophage colony stimulating factor (M-CSF) can act in synergistic way to promote the growth of mononuclear phagocytes. SCF-M-CSF fusion proteins were designed on the computer using the Homology and Biopolymer modules of the software packages InsightII. Several existing crystal structures were used as templates to generate models of the complexes of receptor with fusion protein. The structure rationality of the fusion protein incorporated a series of flexible linker peptide was analyzed on InsightII system. Then, a suitable peptide GGGGSGGGGSGG was chosen for the fusion protein. Two recombinant SCF-M-CSF fusion proteins were generated by construction of a plasmid in which the coding regions of human SCF (1-165aa) and M-CSF (1-149aa) cDNA were connected by this linker peptide coding sequence followed by subsequent expression in insect cell. The results of Western blot and activity analysis showed that these two recombinant fusion proteins existed as a dimer with a molecular weight of 84 KD under non-reducing conditions and a monomer of 42 KD at reducing condition. The results of cell proliferation assays showed that each fusion protein induced a dose-dependent proliferative response. At equimolar concentration, SCF/M-CSF was about 20 times more potent than the standard monomeric SCF in stimulating TF-1 cell line growth, while M-CSF/SCF was 10 times of monomeric SCF. No activity difference of M-CSF/SCF or SCF/M-CSF to M-CSF (at same molar) was found in stimulating the HL-60 cell linear growth. The synergistic effect of SCF and M-CSF moieties in the fusion proteins was demonstrated by the result of clonogenic assay performed with human bone mononuclear, in which both SCF/M-CSF and M-CSF/SCF induced much higher number of CFU-M than equimolar amount of SCF or M-CSF or that of two cytokines mixture.

  6. Fusion target design

    International Nuclear Information System (INIS)

    Bangerter, R.O.

    1978-01-01

    Most detailed fusion target design is done by numerical simulation using large computers. Although numerical simulation is briefly discussed, this lecture deals primarily with the way in which basic physical arguments, driver technology considerations and economical power production requirements are used to guide and augment the simulations. Physics topics discussed include target energetics, preheat, stability and symmetry. A specific design example is discussed

  7. Preparation of GST Fusion Proteins.

    Science.gov (United States)

    Einarson, Margret B; Pugacheva, Elena N; Orlinick, Jason R

    2007-04-01

    INTRODUCTIONThis protocol describes the preparation of glutathione-S-transferase (GST) fusion proteins, which have had a wide range of applications since their introduction as tools for synthesis of recombinant proteins in bacteria. GST was originally selected as a fusion moiety because of several desirable properties. First and foremost, when expressed in bacteria alone, or as a fusion, GST is not sequestered in inclusion bodies (in contrast to previous fusion protein systems). Second, GST can be affinity-purified without denaturation because it binds to immobilized glutathione, which provides the basis for simple purification. Consequently, GST fusion proteins are routinely used for antibody generation and purification, protein-protein interaction studies, and biochemical analysis.

  8. Fusion engineering device design description

    Energy Technology Data Exchange (ETDEWEB)

    Flanagan, C.A.; Steiner, D.; Smith, G.E.

    1981-12-01

    The US Magnetic Fusion Engineering Act of 1980 calls for the operation of a Fusion Engineering Device (FED) by 1990. It is the intent of the Act that the FED, in combination with other testing facilities, will establish the engineering feasibility of magnetic fusion energy. During 1981, the Fusion Engineering Design Center (FEDC), under the guidance of a Technical Management Board (TMB), developed a baseline design for the FED. This design is summarized herein.

  9. Fusion Engineering Device design description

    International Nuclear Information System (INIS)

    Flanagan, C.A.; Steiner, D.; Smith, G.E.

    1981-12-01

    The US Magnetic Fusion Engineering Act of 1980 calls for the operation of a Fusion Engineering Device (FED) by 1990. It is the intent of the Act that the FED, in combination with other testing facilities, will establish the engineering feasibility of magnetic fusion energy. During 1981, the Fusion Engineering Design Center (FEDC), under the guidance of a Technical Management Board (TMB), developed a baseline design for the FED. This design is summarized herein

  10. Fusion engineering device design description

    International Nuclear Information System (INIS)

    Flanagan, C.A.; Steiner, D.; Smith, G.E.

    1981-12-01

    The US Magnetic Fusion Engineering Act of 1980 calls for the operation of a Fusion Engineering Device (FED) by 1990. It is the intent of the Act that the FED, in combination with other testing facilities, will establish the engineering feasibility of magnetic fusion energy. During 1981, the Fusion Engineering Design Center (FEDC), under the guidance of a Technical Management Board (TMB), developed a baseline design for the FED. This design is summarized herein

  11. Cellulose binding domain fusion proteins

    Science.gov (United States)

    Shoseyov, Oded; Shpiegl, Itai; Goldstein, Marc A.; Doi, Roy H.

    1998-01-01

    A cellulose binding domain (CBD) having a high affinity for crystalline cellulose and chitin is disclosed, along with methods for the molecular cloning and recombinant production thereof. Fusion products comprising the CBD and a second protein are likewise described. A wide range of applications are contemplated for both the CBD and the fusion products, including drug delivery, affinity separations, and diagnostic techniques.

  12. Design of Fusion Proteins for Efficient and Soluble Production of Immunogenic Ebola Virus Glycoprotein in Escherichia coli.

    Science.gov (United States)

    Ji, Yang; Lu, Yuan; Yan, Yishu; Liu, Xinxin; Su, Nan; Zhang, Chong; Bi, Shengli; Xing, Xin-Hui

    2018-03-03

    The Ebola hemorrhagic fever caused by Ebola virus is an extremely dangerous disease, and effective therapeutic agents are still lacking. Platforms for the efficient production of vaccines are crucial to ensure quick response against an Ebola virus outbreak. Ebola virus glycoprotein (EbolaGP) on the virion surface is responsible for membrane binding and virus entry, thus becoming the key target for vaccine development. However, heterologous expression of this protein still faces engineering challenges such as low production levels and insoluble aggregation. Here, the authors design and compare various fusion strategies, attaching great importance to the solubility-enhancing effect, and tag removal process. It is found that a C-terminal intein-based tag greatly enhances the solubility of EbolaGP and allows one-step chromatographic purification of the untagged EbolaGP through thiol-catalyzed self-cleavage. The purified untagged EbolaGP alone or with Freund's adjuvant are highly immunogenic, as confirmed in a mouse model. Consequently, the present study puts forward a new strategy for the efficient and soluble expression of untagged immunogenic EbolaGP. The intein-based protein fusion approach may be of importance for the large-scale production of Ebola virus subunit vaccine. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. Inertial fusion reactor designs

    International Nuclear Information System (INIS)

    Meier, W.

    1987-01-01

    In this paper, a variety of reactor concepts are proposed. One of the prime concerns is dealing with the x-rays and debris that are emitted by the target. Internal neutron shielding can reduce radiation damage and activation, leading to longer life systems, reduced activation and fewer safety concerns. There is really no consensus on what the best reactor concept is at this point. There has been virtually no chamber technology development to date. This is the flip side of the coin of the separability of the target physics and the reactor design. Since reactor technology has not been required to do target experiments, it's not being developed. Economic analysis of conceptual designs indicates that ICF can be economically competitive with magnetic fusion, fission and fossil plants

  14. Application of Quality by Design to the characterization of the cell culture process of an Fc-Fusion protein.

    Science.gov (United States)

    Rouiller, Yolande; Solacroup, Thomas; Deparis, Véronique; Barbafieri, Marco; Gleixner, Ralf; Broly, Hervé; Eon-Duval, Alex

    2012-06-01

    The production bioreactor step of an Fc-Fusion protein manufacturing cell culture process was characterized following Quality by Design principles. Using scientific knowledge derived from the literature and process knowledge gathered during development studies and manufacturing to support clinical trials, potential critical and key process parameters with a possible impact on product quality and process performance, respectively, were determined during a risk assessment exercise. The identified process parameters were evaluated using a design of experiment approach. The regression models generated from the data allowed characterizing the impact of the identified process parameters on quality attributes. The main parameters having an impact on product titer were pH and dissolved oxygen, while those having the highest impact on process- and product-related impurities and variants were pH and culture duration. The models derived from characterization studies were used to define the cell culture process design space. The design space limits were set in such a way as to ensure that the drug substance material would consistently have the desired quality. Copyright © 2012 Elsevier B.V. All rights reserved.

  15. Fc-fusion Proteins in Therapy: An Updated View.

    Science.gov (United States)

    Jafari, Reza; Zolbanin, Naime M; Rafatpanah, Houshang; Majidi, Jafar; Kazemi, Tohid

    2017-01-01

    Fc-fusion proteins are composed of Fc region of IgG antibody (Hinge-CH2-CH3) and a desired linked protein. Fc region of Fc-fusion proteins can bind to neonatal Fc receptor (FcRn) thereby rescuing it from degradation. The first therapeutic Fc-fusion protein was introduced for the treatment of AIDS. The molecular designing is the first stage in production of Fc-fusion proteins. The amino acid residues in the Fc region and linked protein are very important in the bioactivity and affinity of the fusion proteins. Although, therapeutic monoclonal antibodies are the top selling biologics but the application of therapeutic Fc-fusion proteins in clinic is in progress and among these medications Etanercept is the most effective in therapy. At present, eleven Fc-fusion proteins have been approved by FDA. There are novel Fc-fusion proteins which are in pre-clinical and clinical development. In this article, we review the molecular and biological characteristics of Fc-fusion proteins and then further discuss the features of novel therapeutic Fc-fusion proteins. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  16. Designing proteins for therapeutic applications.

    Science.gov (United States)

    Lazar, Greg A; Marshall, Shannon A; Plecs, Joseph J; Mayo, Stephen L; Desjarlais, John R

    2003-08-01

    Protein design is becoming an increasingly useful tool for optimizing protein drugs and creating novel biotherapeutics. Recent progress includes the engineering of monoclonal antibodies, cytokines, enzymes and viral fusion inhibitors.

  17. Designing the Sniper: Improving Targeted Human Cytolytic Fusion Proteins for Anti-Cancer Therapy via Molecular Simulation

    Directory of Open Access Journals (Sweden)

    Anna Bochicchio

    2017-02-01

    Full Text Available Targeted human cytolytic fusion proteins (hCFPs are humanized immunotoxins for selective treatment of different diseases including cancer. They are composed of a ligand specifically binding to target cells genetically linked to a human apoptosis-inducing enzyme. hCFPs target cancer cells via an antibody or derivative (scFv specifically binding to e.g., tumor associated antigens (TAAs. After internalization and translocation of the enzyme from endocytosed endosomes, the human enzymes introduced into the cytosol are efficiently inducing apoptosis. Under in vivo conditions such enzymes are subject to tight regulation by native inhibitors in order to prevent inappropriate induction of cell death in healthy cells. Tumor cells are known to upregulate these inhibitors as a survival mechanism resulting in escape of malignant cells from elimination by immune effector cells. Cytosolic inhibitors of Granzyme B and Angiogenin (Serpin P9 and RNH1, respectively, reduce the efficacy of hCFPs with these enzymes as effector domains, requiring detrimentally high doses in order to saturate inhibitor binding and rescue cytolytic activity. Variants of Granzyme B and Angiogenin might feature reduced affinity for their respective inhibitors, while retaining or even enhancing their catalytic activity. A powerful tool to design hCFPs mutants with improved potency is given by in silico methods. These include molecular dynamics (MD simulations and enhanced sampling methods (ESM. MD and ESM allow predicting the enzyme-protein inhibitor binding stability and the associated conformational changes, provided that structural information is available. Such “high-resolution” detailed description enables the elucidation of interaction domains and the identification of sites where particular point mutations may modify those interactions. This review discusses recent advances in the use of MD and ESM for hCFP development from the viewpoints of scientists involved in both fields.

  18. Fluorescent S-layer fusion proteins

    International Nuclear Information System (INIS)

    Kainz, B.

    2010-01-01

    This work describes the construction and characterisation of fluorescent S-layer fusion proteins used as building blocks for the fabrication of nanostructured monomolecular biocoatings on silica particles with defined fluorescence properties. The S-layer protein SgsE of Geobacillus stearothermophilus NRS 2004/3a was fused with the pH-dependant cyan, green and yellow variant of the green fluorescent protein (GFP) and the red fluorescent protein mRFP1. These fluorescent S-layer fusion proteins, acting as scaffold and optical sensing element simultaneously, were able to reassemble in solution and on silica particles forming 2D nanostructures with p2 lattice symmetry (a=11 ±0.5 nm, b=14 ±0.4 nm, g=80 ±1 o ). The pH-dependant fluorescence behaviour was studied with fluorimetry, confocal microscopy and flow cytometry. These fluorescent S-layer fusion proteins can be used as pH-sensor. 50% of the fluorescence intensity decreases at their calculated pKa values (pH6 - pH5). The fluorescence intensity of the GFP variants vanished completely between pH4 and pH3 whereas the chromophore of the red protein mRFP1 was only slightly affected in acidic conditions. At the isoelectric point of the S-layer coated silica particles (pH4.6 ±0.2) an increase in particle aggregation was detected by flow cytometry. The cyan and yellow fluorescent proteins were chosen to create a bi-fluorescent S-layer tandem fusion protein with the possibility for resonance energy transfer (FRET). A transfer efficiency of 20% and a molecular distance between the donor (ECFP) and acceptor (YFP) chromophores of around 6.2 nm could be shown. This bi-fluorescent ECFP-SgsE-YFP tandem fusion protein was able to reassemble on solid surfaces. The remarkable combination of fluorescence and self-assembly and the design of bi-functional S-layer tandem fusion protein matrices makes them to a promising tool in nanobiotechnology. (author) [de

  19. Exo-endo cellulase fusion protein

    Science.gov (United States)

    Bower, Benjamin S [Palo Alto, CA; Larenas, Edmund A [Palo Alto, CA; Mitchinson, Colin [Palo Alto, CA

    2012-01-17

    The present invention relates to a heterologous exo-endo cellulase fusion construct, which encodes a fusion protein having cellulolytic activity comprising a catalytic domain derived from a fungal exo-cellobiohydrolase and a catalytic domain derived from an endoglucanase. The invention also relates to vectors and fungal host cells comprising the heterologous exo-endo cellulase fusion construct as well as methods for producing a cellulase fusion protein and enzymatic cellulase compositions.

  20. Mirror fusion reactor design

    International Nuclear Information System (INIS)

    Neef, W.S. Jr.; Carlson, G.A.

    1979-01-01

    Recent conceptual reactor designs based on mirror confinement are described. Four components of mirror reactors for which materials considerations and structural mechanics analysis must play an important role in successful design are discussed. The reactor components are: (a) first-wall and thermal conversion blanket, (b) superconducting magnets and their force restraining structure, (c) neutral beam injectors, and (d) plasma direct energy converters

  1. Advances in fusion reactor design

    International Nuclear Information System (INIS)

    Baker, C.C.

    1987-01-01

    The author addresses the tokamak as a power reactor. Contrary to popular opinion, there are still a few people that think a tokamak might make a good fusion power reactor. In thinking about advances in fusion reactor design, in the U.S., at least, that generally means advances relevant to the Starfire design. He reviews some of the features of Starfire. Starfire is the last major study done of the tokamak as a reactor in this country. It is now over eight years old in the sense that eight years ago was really the time in which major decisions were made as to its features. Starfire was a tokamak with a major radius of seven meters, about twice the linear dimensions of a machine like TIBER

  2. Design activities of a fusion experimental breeder

    International Nuclear Information System (INIS)

    Huang, J.; Feng, K.; Sheng, G.

    1999-01-01

    The fusion reactor design studies in China are under the support of a fusion-fission hybrid reactor research Program. The purpose of this program is to explore the potential near-term application of fusion energy to support the long-term fusion energy on the one hand and the fission energy development on the other. During 1992-1996 a detailed consistent and integral conceptual design of a Fusion Experimental Breeder, FEB was completed. Beginning from 1996, a further design study towards an Engineering Outline Design of the FEB, FEB-E, has started. The design activities are briefly given. (author)

  3. Design activities of a fusion experimental breeder

    International Nuclear Information System (INIS)

    Huang, J.; Feng, K.; Sheng, G.

    2001-01-01

    The fusion reactor design studies in China are under the support of a fusion-fission hybrid reactor research Program. The purpose of this program is to explore the potential near-term application of fusion energy to support the long-term fusion energy on the one hand and the fission energy development on the other. During 1992-1996 a detailed consistent and integral conceptual design of a Fusion Experimental Breeder, FEB was completed. Beginning from 1996, a further design study towards an Engineering Outline Design of the FEB, FEB-E, has started. The design activities are briefly given. (author)

  4. A design of steady state fusion burner

    International Nuclear Information System (INIS)

    Hasegawa, Akira; Hatori, Tadatsugu; Itoh, Kimitaka; Ikuta, Takashi; Kodama, Yuji.

    1975-01-01

    We present a brief design of a steady state fusion burner in which a continuous burning of nuclear fuel may be achieved with output power of a gigawatt. The laser fusion is proposed to ignite the fuel. (auth.)

  5. Advances in laser solenoid fusion reactor design

    International Nuclear Information System (INIS)

    Steinhauer, L.C.; Quimby, D.C.

    1978-01-01

    The laser solenoid is an alternate fusion concept based on a laser-heated magnetically-confined plasma column. The reactor concept has evolved in several systems studies over the last five years. We describe recent advances in the plasma physics and technology of laser-plasma coupling. The technology advances include progress on first walls, inner magnet design, confinement module design, and reactor maintenance. We also describe a new generation of laser solenoid fusion and fusion-fission reactor designs

  6. Design study of laser fusion rocket

    International Nuclear Information System (INIS)

    Nakashima, Hideki; Shoyama, Hidetoshi; Kanda, Yukinori

    1991-01-01

    A design study was made on a rocket powered by laser fusion. Dependence of its flight performance on target gain, driver repetition rate and fuel composition was analyzed to obtain optimal design parameters of the laser fusion rocket. The results indicate that the laser fusion rocket fueled with DT or D 3 He has the potential advantages over other propulsion systems such as fission rocket for interplanetary travel. (author)

  7. Mitochondrial Fusion Proteins and Human Diseases

    Directory of Open Access Journals (Sweden)

    Michela Ranieri

    2013-01-01

    Full Text Available Mitochondria are highly dynamic, complex organelles that continuously alter their shape, ranging between two opposite processes, fission and fusion, in response to several stimuli and the metabolic demands of the cell. Alterations in mitochondrial dynamics due to mutations in proteins involved in the fusion-fission machinery represent an important pathogenic mechanism of human diseases. The most relevant proteins involved in the mitochondrial fusion process are three GTPase dynamin-like proteins: mitofusin 1 (MFN1 and 2 (MFN2, located in the outer mitochondrial membrane, and optic atrophy protein 1 (OPA1, in the inner membrane. An expanding number of degenerative disorders are associated with mutations in the genes encoding MFN2 and OPA1, including Charcot-Marie-Tooth disease type 2A and autosomal dominant optic atrophy. While these disorders can still be considered rare, defective mitochondrial dynamics seem to play a significant role in the molecular and cellular pathogenesis of more common neurodegenerative diseases, for example, Alzheimer’s and Parkinson’s diseases. This review provides an overview of the basic molecular mechanisms involved in mitochondrial fusion and focuses on the alteration in mitochondrial DNA amount resulting from impairment of mitochondrial dynamics. We also review the literature describing the main disorders associated with the disruption of mitochondrial fusion.

  8. High temperature fusion reactor design

    International Nuclear Information System (INIS)

    Harkness, S.D.; dePaz, J.F.; Gohar, M.Y.; Stevens, H.C.

    1979-01-01

    Fusion energy may have unique advantages over other systems as a source for high temperature process heat. A conceptual design of a blanket for a 7 m tokamak reactor has been developed that is capable of producing 1100 0 C process heat at a pressure of approximately 10 atmospheres. The design is based on the use of a falling bed of MgO spheres as the high temperature heat transfer system. By preheating the spheres with energy taken from the low temperature tritium breeding part of the blanket, 1086 MW of energy can be generated at 1100 0 C from a system that produces 3000 MW of total energy while sustaining a tritium breeding ratio of 1.07. The tritium breeding is accomplished using Li 2 O modules both in front of (6 cm thick) and behind (50 cm thick) the high temperature ducts. Steam is used as the first wall and front tritium breeding module coolant while helium is used in the rear tritium breeding region. The system produces 600 MW of net electricity for use on the grid

  9. Fluorescent sensors based on bacterial fusion proteins

    International Nuclear Information System (INIS)

    Mateu, Batirtze Prats; Pum, Dietmar; Sleytr, Uwe B; Toca-Herrera, José L; Kainz, Birgit

    2014-01-01

    Fluorescence proteins are widely used as markers for biomedical and technological purposes. Therefore, the aim of this project was to create a fluorescent sensor, based in the green and cyan fluorescent protein, using bacterial S-layers proteins as scaffold for the fluorescent tag. We report the cloning, expression and purification of three S-layer fluorescent proteins: SgsE-EGFP, SgsE-ECFP and SgsE-13aa-ECFP, this last containing a 13-amino acid rigid linker. The pH dependence of the fluorescence intensity of the S-layer fusion proteins, monitored by fluorescence spectroscopy, showed that the ECFP tag was more stable than EGFP. Furthermore, the fluorescent fusion proteins were reassembled on silica particles modified with cationic and anionic polyelectrolytes. Zeta potential measurements confirmed the particle coatings and indicated their colloidal stability. Flow cytometry and fluorescence microscopy showed that the fluorescence of the fusion proteins was pH dependent and sensitive to the underlying polyelectrolyte coating. This might suggest that the fluorescent tag is not completely exposed to the bulk media as an independent moiety. Finally, it was found out that viscosity enhanced the fluorescence intensity of the three fluorescent S-layer proteins. (paper)

  10. Design and production of various fusion proteins of the nicotinamide/nicotinate mononucleotide adenilil transferase (NMNAT of Plasmodium falciparum

    Directory of Open Access Journals (Sweden)

    Carlos Alfonso Nieto Clavijo

    2017-09-01

    Full Text Available Recombinant proteins have become useful tools in biochemistry research. During their production, however, inclusion bodies (IB appear, on the one hand, due to the high expression rate from the recombinant plasmids, which have high efficiency promoters, and, on the other hand, intrinsic characteristics of the expressed protein. Furhtermore, the nicotinamide/nicotinate mononucleotide adenilyl transferase (NMNAT is a central protein in NAD(H+ biosynthesis, an essential cofactor in cell metabolism, and in protozoon parasite has been studied. To study the NMNAT protein of these parasites, their recombinant version in E. coli has been expressed, getting a great quantity of IB as a by-product. To increase the solubility of the protein, the coding sequence of the NMNAT enzyme of Plasmodium falciparum was cloned in different expression plasmids which were subsequently transformed into E. coli BL21(DE3 expression strain. The solubility of the recombinant proteins was assessed and the one with the highest presence in the soluble fraction was subsequently purified and its enzyme activity was determined. The recombinant protein with a MBP (maltose-binding protein tag showed an increased solubility and purity.

  11. Fusion proteins useful for producing pinene

    Energy Technology Data Exchange (ETDEWEB)

    Peralta-Yahya, Pamela P.; Keasling, Jay D

    2016-06-28

    The present invention provides for a modified host cell comprising a heterologous pinene synthase (PS), or enzymatically active fragment or variant thereof, and optionally a geranyl pyrophosphate synthase (GPPS), or enzymatically active fragment or variant thereof, or a fusion protein comprising: (a) a PS and (b) a GPPS linked by a linker.

  12. Health physics in fusion reactor design

    International Nuclear Information System (INIS)

    Wong, K.Y.; Dinner, P.J.

    1984-06-01

    Experience in the control of tritium exposures to workers and the public gained through the design and operation of Ontario Hydro's nuclear stations has been applied to fusion projects and to design studies on emerging fusion reactor concepts. Ontario Hydro performance in occupational tritium exposure control and environmental impact is reviewed. Application of tritium control technologies and dose management methodology during facility design is highlighted

  13. Fusion Engineering Device. Volume II. Design description

    International Nuclear Information System (INIS)

    1981-10-01

    This volume summarizes the design of the FED. It includes a description of the major systems and subsystems, the supporting plasma design analysis, a projected device cost and associated construction schedule, and a description of the facilities to house and support the device. This effort represents the culmination of the FY81 studies conducted at the Fusion Engineering Design Center (FEDC). Unique in these design activities has been the collaborative involvement of the Design Center personnel and numerous resource physicists from the fusion community who have made significant contributions in the physics design analysis as well as the physics support of the engineering design of the major FED systems and components

  14. SOLASE: a conceptual laser fusion reactor design

    International Nuclear Information System (INIS)

    Conn, R.W.; Abdel-Khalik, S.I.; Moses, G.A.

    1977-12-01

    The SOLASE conceptual laser fusion reactor has been designed to elucidate the technological problems posed by inertial confinement fusion reactors. This report contains a detailed description of all aspects of the study including the physics of pellet implosion and burn, optics and target illumination, last mirror design, laser system analysis, cavity design, pellet fabrication and delivery, vacuum system requirements, blanket design, thermal hydraulics, tritium analysis, neutronics calculations, radiation effects, stress analysis, shield design, reactor and plant building layout, maintenance procedures, and power cycle design. The reactor is designed as a 1000 MW/sub e/ unit for central station electric power generation

  15. SOLASE: a conceptual laser fusion reactor design

    International Nuclear Information System (INIS)

    Conn, R.W.; Abdel-Khalik, S.I.; Moses, G.A.

    1977-12-01

    The SOLASE conceptual laser fusion reactor has been designed to elucidate the technological problems posed by inertial confinement fusion ractors. This report contains a detailed description of all aspects of the study including the physics of pellet implosion and burn, optics and target illumination, last mirror design, laser system analysis, cavity design, pellet fabrication and delivery, vacuum system requirements, blanket design, thermal hydraulics, tritium analysis, neutronics calculations, radiation effects, stress analysis, shield design, reactor and plant building layout, maintenance procedures, and power cycle design. The reactor is designed as a 1000 MW/sub e/ unit for central station electric power generation

  16. Identification of Protein-Protein Interactions with Glutathione-S-Transferase (GST) Fusion Proteins.

    Science.gov (United States)

    Einarson, Margret B; Pugacheva, Elena N; Orlinick, Jason R

    2007-08-01

    INTRODUCTIONGlutathione-S-transferase (GST) fusion proteins have had a wide range of applications since their introduction as tools for synthesis of recombinant proteins in bacteria. GST was originally selected as a fusion moiety because of several desirable properties. First and foremost, when expressed in bacteria alone, or as a fusion, GST is not sequestered in inclusion bodies (in contrast to previous fusion protein systems). Second, GST can be affinity-purified without denaturation because it binds to immobilized glutathione, which provides the basis for simple purification. Consequently, GST fusion proteins are routinely used for antibody generation and purification, protein-protein interaction studies, and biochemical analysis. This article describes the use of GST fusion proteins as probes for the identification of protein-protein interactions.

  17. A novel TNFα antagonizing peptide-Fc fusion protein designed based on CDRs of TNFα neutralizing monoclonal antibody

    International Nuclear Information System (INIS)

    Qin Weisong; Feng Jiannan; Zhang Wei; Li Yan; Shen, Beifen

    2004-01-01

    The variable regions of antibody molecules bind antigens with high affinity and specificity. The binding sites are imparted largely to the hypervariable portions (i.e., CDRs) of the variable region. Peptides derived from CDRs can bind antigen with similar specificity acting as mimic of antibody and become drug-designing core, although with markedly lower affinity. In order to increase the affinity and bioactivity, in this study, a novel peptide (PT) designed on CDRs of a TNFα neutralizing monoclonal antibody Z12 was linked with Fc fragment of human IgG1. The interaction mode of PT-linker-Fc (PLF) with TNFα was analyzed with computer-guided molecular modeling method. After expression in Escherichia coli and purification, recombinant PT-linker-Fc could bind directly with the TNFα coated on the ELISA plates. Furthermore, PLF could competitively inhibit the binding of Z12 to TNFα and also inhibit the TNFα-induced cytotoxicity on L929 cells. The TNFα antagonizing activity of PLF was significantly higher than that of the free peptide. This study highlights the potential of human Fc to enhance the potency of peptides designed on the CDRs of antibodies and could be useful in developing new TNFα antagonists

  18. Introduction to magnetic fusion reactor design

    International Nuclear Information System (INIS)

    Watanabe, Kenji

    1988-01-01

    Trend of the tokamak reactor design works so far carried out is reviewed, and method of conceptual design for commercial fusion reactor is critically considered concerning the black-box conpepts. System-framework of the engineering of magnetic fusion (commercial) reactor design is proposed as four steps. Based on it the next design studies are recommended in parallel approaches for making real-overcome of reactor material problem, from the view point of technological realization and not from the economical one. Real trials are involved. (author)

  19. Current fusion power plant design concepts

    International Nuclear Information System (INIS)

    Gore, B.F.; Murphy, E.S.

    1976-09-01

    Nine current U.S. designs for fusion power plants are described in this document. Summary tabulations include a tenth concept, for which the design document was unavailable during preparation of the descriptions. The information contained in the descriptions was used to define an envelope of fusion power plant characteristics which formed the basis for definition of reference first commercial fusion power plant design. A brief prose summary of primary plant features introduces each of the descriptions contained in the body of this document. In addition, summary tables are presented. These tables summarize in side-by-side fashion, plant parameters, processes, combinations of materials used, requirements for construction materials, requirements for replacement materials during operation, and production of wastes

  20. General software design for multisensor data fusion

    Science.gov (United States)

    Zhang, Junliang; Zhao, Yuming

    1999-03-01

    In this paper a general method of software design for multisensor data fusion is discussed in detail, which adopts object-oriented technology under UNIX operation system. The software for multisensor data fusion is divided into six functional modules: data collection, database management, GIS, target display and alarming data simulation etc. Furthermore, the primary function, the components and some realization methods of each modular is given. The interfaces among these functional modular relations are discussed. The data exchange among each functional modular is performed by interprocess communication IPC, including message queue, semaphore and shared memory. Thus, each functional modular is executed independently, which reduces the dependence among functional modules and helps software programing and testing. This software for multisensor data fusion is designed as hierarchical structure by the inheritance character of classes. Each functional modular is abstracted and encapsulated through class structure, which avoids software redundancy and enhances readability.

  1. Impact of fluorescent protein fusions on the bacterial flagellar motor

    NARCIS (Netherlands)

    Heo, M.; Nord, A. L.; Chamousset, D.; van Rijn, E.; Beaumont, H.J.E.; Pedaci, F.

    2017-01-01

    Fluorescent fusion proteins open a direct and unique window onto protein function. However, they also introduce the risk of perturbation of the function of the native protein. Successful applications of fluorescent fusions therefore rely on a careful assessment and minimization of the side

  2. Designing the Cascade inertial confinement fusion reactor

    International Nuclear Information System (INIS)

    Pitts, J.H.

    1987-01-01

    The primary goal in designing inertial confinement fusion (ICF) reactors is to produce electrical power as inexpensively as possible, with minimum activation and without compromising safety. This paper discusses a method for designing the Cascade rotating ceramic-granule-blanket reactor (Pitts, 1985) and its associated power plant (Pitts and Maya, 1985). Although focus is on the cascade reactor, the design method and issues presented are applicable to most other ICF reactors

  3. Materials design data for fusion reactors

    International Nuclear Information System (INIS)

    Tavassoli, A.A.F.

    1998-01-01

    Design data needed for fusion reactors are characterized by the diversity of materials and the complexity of loading situations found in these reactors. In addition, advanced fabrication techniques, such as hot isostatic pressing, envisaged for fabrication of single and multilayered in-vessel components, could significantly change the original materials properties for which the current design rules are written. As a result, additional materials properties have had to be generated for fusion reactors and new structural design rules formulated. This paper recalls some of the materials properties data generated for ITER and DEMO, and gives examples of how these are converted into design criteria. In particular, it gives specific examples for the properties of 316LN-IG and modified 9Cr-1Mo steels, and CuCrZr alloy. These include, determination of tension, creep, isochronous, fatigue, and creep-fatigue curves and their analysis and conversion into design limits. (orig.)

  4. Materials design data for fusion reactors

    Energy Technology Data Exchange (ETDEWEB)

    Tavassoli, A.A.F. [CEA Commissariat a l`Energie Atomique, Gif sur Yvette (France). CEREM

    1998-10-01

    Design data needed for fusion reactors are characterized by the diversity of materials and the complexity of loading situations found in these reactors. In addition, advanced fabrication techniques, such as hot isostatic pressing, envisaged for fabrication of single and multilayered in-vessel components, could significantly change the original materials properties for which the current design rules are written. As a result, additional materials properties have had to be generated for fusion reactors and new structural design rules formulated. This paper recalls some of the materials properties data generated for ITER and DEMO, and gives examples of how these are converted into design criteria. In particular, it gives specific examples for the properties of 316LN-IG and modified 9Cr-1Mo steels, and CuCrZr alloy. These include, determination of tension, creep, isochronous, fatigue, and creep-fatigue curves and their analysis and conversion into design limits. (orig.) 19 refs.

  5. Recombinant fusion protein of albumin-retinol binding protein inactivates stellate cells

    International Nuclear Information System (INIS)

    Choi, Soyoung; Park, Sangeun; Kim, Suhyun; Lim, Chaeseung; Kim, Jungho; Cha, Dae Ryong; Oh, Junseo

    2012-01-01

    Highlights: ► We designed novel recombinant albumin-RBP fusion proteins. ► Expression of fusion proteins inactivates pancreatic stellate cells (PSCs). ► Fusion proteins are successfully internalized into and inactivate PSCs. ► RBP moiety mediates cell specific uptake of fusion protein. -- Abstract: Quiescent pancreatic- (PSCs) and hepatic- (HSCs) stellate cells store vitamin A (retinol) in lipid droplets via retinol binding protein (RBP) receptor and, when activated by profibrogenic stimuli, they transform into myofibroblast-like cells which play a key role in the fibrogenesis. Despite extensive investigations, there is, however, currently no appropriate therapy available for tissue fibrosis. We previously showed that the expression of albumin, composed of three homologous domains (I–III), inhibits stellate cell activation, which requires its high-affinity fatty acid-binding sites asymmetrically distributed in domain I and III. To attain stellate cell-specific uptake, albumin (domain I/III) was coupled to RBP; RBP-albumin domain III (R-III) and albumin domain I -RBP-albumin III (I-R-III). To assess the biological activity of fusion proteins, cultured PSCs were used. Like wild type albumin, expression of R-III or I-R-III in PSCs after passage 2 (activated PSCs) induced phenotypic reversal from activated to fat-storing cells. On the other hand, R-III and I-R-III, but not albumin, secreted from transfected 293 cells were successfully internalized into and inactivated PSCs. FPLC-purified R-III was found to be internalized into PSCs via caveolae-mediated endocytosis, and its efficient cellular uptake was also observed in HSCs and podocytes among several cell lines tested. Moreover, tissue distribution of intravenously injected R-III was closely similar to that of RBP. Therefore, our data suggest that albumin-RBP fusion protein comprises of stellate cell inactivation-inducing moiety and targeting moiety, which may lead to the development of effective anti

  6. Pellet design for a laser fusion reactor

    International Nuclear Information System (INIS)

    Thiessen, A.R.; Nuckolls, J.

    1974-01-01

    The requirements for laser fusion pellet design are discussed. Computer calculations are presented of a capsule consisting of a spherical solid drop of DT surrounded by a concentric shell of DT. Gains greater than 40 fold are achieved with laser energies of approximately 0.5 MJ, and peak powers of about 10 16 W. (U.S.)

  7. Scyllac fusion test reactor design

    International Nuclear Information System (INIS)

    Dudziak, D.J.; Gerstl, S.A.; Houck, D.L.; Jalbert, R.A.; Krakowski, R.A.; Linford, R.K.; McDonald, T.E.; Rogers, J.D.; Thomassen, K.I.

    1975-01-01

    A general design of the system is given. The implosion heating and compression systems (METS) are described. Tritium handling, shielding and activation of the reactor, and safety and environmental aspects are discussed

  8. Status of fusion reactor blanket design

    International Nuclear Information System (INIS)

    Smith, D.L.; Sze, D.K.

    1986-02-01

    The recent Blanket Comparison and Selection Study (BCSS), which was a comprehensive evaluation of fusion reactor blanket design and the status of blanket technology, serves as an excellent basis for further development of blanket technology. This study provided an evaluation of over 130 blanket concepts for the reference case of electric power producing, DT fueled reactors in both Tokamak and Tandem Mirror (TMR) configurations. Based on a specific set of reactor operating parameters, the current understanding of materials and blanket technology, and a uniform evaluation methodology developed as part of the study, a limited number of concepts were identified that offer the greatest potential for making fusion an attractive energy source

  9. Scyllac fusion test reactor design

    International Nuclear Information System (INIS)

    Linford, R.K.; Oliphant, T.A.; Thomassen, K.I.

    1976-01-01

    The SFTR is a proposed 80-m diameter D-T burning toroidal theta pinch. The system is designed to achieve Q = 1 where Q is the ratio of the total thermonuclear energy output to the maximum stored energy in the plasma. SFTR design studies [1] will provide valuable guidance to the Scyllac related research and to the needed technological development. The portion of the system directly related to the plasma confinement, stability, and heating, is described, and the approach used to obtain an operating point consistent with Q = 1, m = 1 stability, and technological limitations is outlined. (U.K.)

  10. Scyllac fusion test reactor design

    International Nuclear Information System (INIS)

    Linford, R.K.; Oliphant, T.A.; Thomassen, K.I.

    1975-01-01

    The SFTR is a proposed 80-m diameter D--T burning toroidal theta pinch. The system is designed to achieve Q = 1 where Q is the ratio of the total thermonuclear energy output to the maximum stored energy in the plasma. SFTR design studies will provide valuable guidance to the Scyllac related research and to the needed technological development. This paper describes the portion of the system directly related to the plasma confinement, stability, and heating, and outlines the approach used to obtain an operating point consistent with Q = 1, m = 1 stability, and technological limitations. (auth)

  11. Conceptual design of Fusion Experimental Reactor (FER)

    International Nuclear Information System (INIS)

    Tone, T.; Fujisawa, N.

    1983-01-01

    Conceptual design studies of the Fusion Experimental Reactor (FER) have been performed. The FER has an objective of achieving selfignition and demonstrating engineering feasibility as a next generation tokamak to JT-60. Various concepts of the FER have been considered. The reference design is based on a double-null divertor. Optional design studies with some attractive features based on advanced concepts such as pumped limiter and RF current drive have been carried out. Key design parameters are; fusion power of 440 MW, average neutron wall loading of 1MW/m 2 , major radius of 5.5m, plasma minor radius of 1.1m, plasma elongation of 1.5, plasma current of 5.3MA, toroidal beta of 4%, toroidal field on plasma axis of 5.7T and tritium breeding ratio of above unity

  12. Extrap conceptual fusion reactor design study

    International Nuclear Information System (INIS)

    Eninger, J.E; Lehnert, B.

    1987-12-01

    A study has recently been initiated to asses the fusion reactor potential of the Extrap concept. A reactor model is defined that fulfills certain economic and environmental criteria. This model is applied to Extrap and a reference reactor is outlined. The design is optimized by varying parameters subject to both physics and engineering constraints. Several design options are examined and key engineering issues are identified and addressed. Some preliminary results and conclusions of this work are summarized. (authors)

  13. Laser solenoid fusion--fission design

    International Nuclear Information System (INIS)

    Steinhauer, L.C.; Taussig, R.T.

    1976-01-01

    The dependence of breeding performance on system engineering parameters is examined for laser solenoid fusion-fission reactors. Reactor performance is found to be relatively insensitive to most of the engineering parameters, and compact designs can be built based on reasonable technologies. Point designs are described for the prototype series of reactors (mid-term technologies) and for second generation systems (advanced technologies). It is concluded that the laser solenoid has a good probability of timely application to fuel breeding needs

  14. MEMBRANE-FUSION OF SEMLIKI FOREST VIRUS INVOLVES HOMOTRIMERS OF THE FUSION PROTEIN

    NARCIS (Netherlands)

    WAHLBERG, JM; WILSCHUT, J; GAROFF, H

    1992-01-01

    Infection of cells with enveloped viruses is accomplished through membrane fusion. The binding and fusion Processes are mediated by the spike proteins in the envelope of the virus particle and usually involve a series of conformational changes in these proteins. We have studied the low-pH-mediated

  15. Magnet design considerations for Tokamak fusion reactors

    International Nuclear Information System (INIS)

    Purcell, J.R.; Chen, W.; Thomas, R.

    1976-01-01

    Design problems for superconducting ohmic heating and toroidal field coils for large Tokamak fusion reactors are discussed. The necessity for making these coils superconducting is explained, together with the functions of these coils in a Tokamak reactor. Major problem areas include materials related aspects and mechanical design and cryogenic considerations. Projections and comparisons are made based on existing superconducting magnet technology. The mechanical design of large-scale coils, which can contain the severe electromagnetic loading and stress generated in the winding, are emphasized. Additional major tasks include the development of high current conductors for pulsed applications to be used in fabricating the ohmic heating coils. It is important to note, however, that no insurmountable technical barriers are expected in the course of developing superconducting coils for Tokamak fusion reactors. (Auth.)

  16. Economic comparison of fusion power plant designs

    International Nuclear Information System (INIS)

    O'Neill, J.E.

    1986-01-01

    Over the past 10 yr, a number of studies have been developed for fusion power plants of various types (tokamaks, mirrors, etc.) complete with figures of merit such as cost estimates and estimates of the cost of generating electricity (COE). Each of these designs involves unresolved physics and engineering problems which, it is assumed, will eventually be worked out. Because of such uncertainties the figures of merit associated with such designs are not to be compared as absolute measures of worth but as relative indicators of progress within a given concept type. As part of Grumman's involvement in fusion energy development, an effort has been undertaken to compare economic indicators from the referenced studies in order to determine the cost trend in recent reactor design activities

  17. Paramyxovirus F1 protein has two fusion peptides: implications for the mechanism of membrane fusion.

    Science.gov (United States)

    Peisajovich, S G; Samuel, O; Shai, Y

    2000-03-10

    Viral fusion proteins contain a highly hydrophobic segment, named the fusion peptide, which is thought to be responsible for the merging of the cellular and viral membranes. Paramyxoviruses are believed to contain a single fusion peptide at the N terminus of the F1 protein. However, here we identified an additional internal segment in the Sendai virus F1 protein (amino acids 214-226) highly homologous to the fusion peptides of HIV-1 and RSV. A synthetic peptide, which includes this region, was found to induce membrane fusion of large unilamellar vesicles, at concentrations where the known N-terminal fusion peptide is not effective. A scrambled peptide as well as several peptides from other regions of the F1 protein, which strongly bind to membranes, are not fusogenic. The functional and structural characterization of this active segment suggest that the F1 protein has an additional internal fusion peptide that could participate in the actual fusion event. The presence of homologous regions in other members of the same family suggests that the concerted action of two fusion peptides, one N-terminal and the other internal, is a general feature of paramyxoviruses. Copyright 2000 Academic Press.

  18. Structural characterization of Mumps virus fusion protein core

    International Nuclear Information System (INIS)

    Liu Yueyong; Xu Yanhui; Lou Zhiyong; Zhu Jieqing; Hu Xuebo; Gao, George F.; Qiu Bingsheng; Rao Zihe; Tien, Po

    2006-01-01

    The fusion proteins of enveloped viruses mediating the fusion between the viral and cellular membranes comprise two discontinuous heptad repeat (HR) domains located at the ectodomain of the enveloped glycoproteins. The crystal structure of the fusion protein core of Mumps virus (MuV) was determined at 2.2 A resolution. The complex is a six-helix bundle in which three HR1 peptides form a central highly hydrophobic coiled-coil and three HR2 peptides pack against the hydrophobic grooves on the surface of central coiled-coil in an oblique antiparallel manner. Fusion core of MuV, like those of simian virus 5 and human respiratory syncytium virus, forms typical 3-4-4-4-3 spacing. The similar charecterization in HR1 regions, as well as the existence of O-X-O motif in extended regions of HR2 helix, suggests a basic rule for the formation of the fusion core of viral fusion proteins

  19. Optical design considerations for laser fusion reactors

    International Nuclear Information System (INIS)

    Monsler, M.J.; Maniscalco, J.A.

    1977-09-01

    The plan for the development of commercial inertial confinement fusion (ICF) power plants is discussed, emphasizing the utilization of the unique features of laser fusion to arrive at conceptual designs for reactors and optical systems which minimize the need for advanced materials and techniques requiring expensive test facilities. A conceptual design for a liquid lithium fall reactor is described which successfully deals with the hostile x-ray and neutron environment and promises to last the 30 year plant lifetime. Schemes for protecting the final focusing optics are described which are both compatible with this reactor system and show promise of surviving a full year in order to minimize costly downtime. Damage mechanisms and protection techniques are discussed, and a recommendation is made for a high f-number metal mirror final focusing system

  20. Fusion breeder sphere - PAC blanket design

    International Nuclear Information System (INIS)

    Sullivan, J.D.; Palmer, B.J.F.

    1987-11-01

    There is a considerable world-wide effort directed toward the production of materials for fusion reactors. Many ceramic fabrication groups are working on making lithium ceramics in a variety of forms, to be incorporated into the tritium breeding blanket which will surround the fusion reactor. Current blanket designs include ceramic in either monolithic or packed sphere bed (sphere-pac) forms. The major thrust at AECL is the production of lithium aluminate spheres to be incorporated in a sphere-pac bed. Contemporary studies on breeder blanket design offer little insight into the requirements on the sizes of the spheres. This study examined the parameters which determine the properties of pressure drop and coolant requirements. It was determined that an optimised sphere-pac bed would be composed of two diameters of spheres: 75 weight % at 3 mm and 25 weight % at 0.3 mm

  1. Recent designs for advanced fusion reactor blankets

    International Nuclear Information System (INIS)

    Sze, D.K.

    1994-06-01

    A series of reactor design studies based on the Tokamak configuration have been carried out under the direction of Professor Robert Conn of UCLA. They are called ARIES-1 through 4 and PULSAR 1 and 2. The key mission of these studies is to evaluate the attractiveness of fusion assuming different degrees of advancement in either physics or engineering development. Also, the requirements of engineering and physics systems for a pulsed reactor were evaluated by the PULSAR design studies. This paper discusses the directions and conclusions of the blanket and related engineering systems for those design studies

  2. Standard mirror fusion reactor design study

    International Nuclear Information System (INIS)

    Moir, R.W.

    1978-01-01

    This report covers the work of the Magnetic Fusion Energy Division's reactor study group during FY 1976 on the standard mirror reactor. The ''standard'' mirror reactor is characterized as a steady state, neutral beam sustained, D-T fusioning plasma confined by a Yin-Yang magnetic mirror field. The physics parameters are obtained from the same physics model that explains the 2XIIB experiment. The model assumes that the drift cyclotron loss cone mode occurs on the boundary of the plasma, and that it is stabilized by warm plasma with negligible energy investment. The result of the study was a workable mirror fusion power plant, steady-state blanket removal made relatively simple by open-ended geometry, and no impurity problem due to the positive plasma potential. The Q (fusion power/injected beam power) turns out to be only 1.1 because of loss out the ends from Coulomb collisions, i.e., classical losses. This low Q resulted in 77% of the gross electrical power being used to power the injectors, thereby causing the net power cost to be high. The low Q stimulated an intensive search for Q-enhancement concepts, resulting in the LLL reactor design effort turning to the field reversal mirror and the tandem mirror, each having Q of order 5

  3. Designs of tandem-mirror fusion reactors

    International Nuclear Information System (INIS)

    Carlson, G.A.; Barr, W.L.; Boghosian, B.M.

    1981-01-01

    We have completed a comparative evaluation of several end plug configurations for tandem mirror fusion reactors with thermal barriers. The axi-cell configuration has been selected for further study and will be the basis for a detailed conceptual design study to be carried out over the next two years. The axi-cell end plug has a simple mirror cell produced by two circular coils followed by a transition coil and a yin-yang pair, which provides for MHD stability

  4. Computational Protein Design

    DEFF Research Database (Denmark)

    Johansson, Kristoffer Enøe

    Proteins are the major functional group of molecules in biology. The impact of protein science on medicine and chemical productions is rapidly increasing. However, the greatest potential remains to be realized. The fi eld of protein design has advanced computational modeling from a tool of support...... to a central method that enables new developments. For example, novel enzymes with functions not found in natural proteins have been de novo designed to give enough activity for experimental optimization. This thesis presents the current state-of-the-art within computational design methods together...... with a novel method based on probability theory. With the aim of assembling a complete pipeline for protein design, this work touches upon several aspects of protein design. The presented work is the computational half of a design project where the other half is dedicated to the experimental part...

  5. Design of a fusion engineering test facility

    International Nuclear Information System (INIS)

    Sager, P.H.

    1980-01-01

    The fusion Engineering Test Facility (ETF) is being designed to provide for engineering testing capability in a program leading to the demonstration of fusion as a viable energy option. It will combine power-reactor-type components and subsystems into an integrated tokamak system and provide a test bed to test blanket modules in a fusion environment. Because of the uncertainties in impurity control two basic designs are being developed: a design with a bundle divertor (Design 1) and one with a poloidal divertor (Design 2). The two designs are similar where possible, the latter having somewhat larger toroidal field (TF) coils to accommodate removal of the larger torus sectors required for the single-null poloidal divertor. Both designs have a major radius of 5.4 m, a minor radius of 1.3 m, and a D-shaped plasma with an elongation of 1.6. Ten TF coils are incorporated in both designs, producing a toroidal field of 5.5 T on-axis. The ohmic heating and equilibrium field (EF) coils supply sufficient volt-seconds to produce a flat-top burn of 100 s and a duty cycle of 135 s, including a start of 12 s, a burn termination of 10 s, and a pumpdown of 13 s. The total fusion power during burn is 750 MW, giving a neutron wall loading of 1.5 MW/m 2 . In Design 1 of the poloidal field (PF) coils except the fast-response EF coils are located outside the FT coils and are superconducting. The fast-response coils are located inside the TF coil bore near the torus and are normal conducting so that they can be easily replaced.In Design 2 all of the PF coils are located outside the TF coils and are superconducting. Ignition is achieved with 60 MW of neutral beam injection at 150 keV. Five megawatts of radio frequency heating (electron cyclotron resonance heating) is used to assist in the startup and limit the breakdown requirement to 25 V

  6. Distinct roles for key karyogamy proteins during yeast nuclear fusion.

    Science.gov (United States)

    Melloy, Patricia; Shen, Shu; White, Erin; Rose, Mark D

    2009-09-01

    During yeast mating, cell fusion is followed by the congression and fusion of the two nuclei. Proteins required for nuclear fusion are found at the surface (Prm3p) and within the lumen (Kar2p, Kar5p, and Kar8p) of the nuclear envelope (NE). Electron tomography (ET) of zygotes revealed that mutations in these proteins block nuclear fusion with different morphologies, suggesting that they act in different steps of fusion. Specifically, prm3 zygotes were blocked before formation of membrane bridges, whereas kar2, kar5, and kar8 zygotes frequently contained them. Membrane bridges were significantly larger and occurred more frequently in kar2 and kar8, than in kar5 mutant zygotes. The kinetics of NE fusion in prm3, kar5, and kar8 mutants, measured by live-cell fluorescence microscopy, were well correlated with the size and frequency of bridges observed by ET. However the kar2 mutant was defective for transfer of NE lumenal GFP, but not diffusion within the lumen, suggesting that transfer was blocked at the NE fusion junction. These observations suggest that Prm3p acts before initiation of outer NE fusion, Kar5p may help dilation of the initial fusion pore, and Kar2p and Kar8p act after outer NE fusion, during inner NE fusion.

  7. Fusion blanket design and optimization techniques

    International Nuclear Information System (INIS)

    Gohar, Y.

    2005-01-01

    In fusion reactors, the blanket design and its characteristics have a major impact on the reactor performance, size, and economics. The selection and arrangement of the blanket materials, dimensions of the different blanket zones, and different requirements of the selected materials for a satisfactory performance are the main parameters, which define the blanket performance. These parameters translate to a large number of variables and design constraints, which need to be simultaneously considered in the blanket design process. This represents a major design challenge because of the lack of a comprehensive design tool capable of considering all these variables to define the optimum blanket design and satisfying all the design constraints for the adopted figure of merit and the blanket design criteria. The blanket design techniques of the First Wall/Blanket/Shield Design and Optimization System (BSDOS) have been developed to overcome this difficulty and to provide the state-of-the-art techniques and tools for performing blanket design and analysis. This report describes some of the BSDOS techniques and demonstrates its use. In addition, the use of the optimization technique of the BSDOS can result in a significant blanket performance enhancement and cost saving for the reactor design under consideration. In this report, examples are presented, which utilize an earlier version of the ITER solid breeder blanket design and a high power density self-cooled lithium blanket design for demonstrating some of the BSDOS blanket design techniques

  8. Conceptual design of fusion experimental reactor (FER)

    International Nuclear Information System (INIS)

    1984-03-01

    A conceptual design study (option C) has been carried out for the fusion experimental reactor (FER). In addition to design of the tokamak reactor and associated systems based on the reference design specifications, feasibility of a water-shield reactor concept was examined as a topical study. The design study for the reference tokamak reactor has produced a reactor concept for the FER, along with major R D items for the concept, based on close examinations on thermal design, electromagnetics, neutronics and remote maintenance. Particular efforts have been directed to the area of electromagnetics. Detailed analyses with close simulation models have been performed on PF coil arrangements and configurations, shell effects of the blanket for plasma position unstability, feedback control, and eddy currents during disruptions. The major design specifications are as follows; Peak fusion power 437 MW Major radius 5.5 m Minor radius 1.1 m Plasma elongation 1.5 Plasma current 5.3 MA Toroidal beta 4 % Field on axis 5.7 T (author)

  9. Conceptual design of fusion experimental reactor (FER)

    International Nuclear Information System (INIS)

    1984-02-01

    This report describes the engineering conceptual design of Fusion Experimental Reactor (FER) which is to be built as a next generation tokamak machine. This design covers overall reactor systems including MHD equilibrium analysis, mechanical configuration of reactor, divertor, pumped limiter, first wall/breeding blanket/shield, toroidal field magnet, poloidal field magnet, cryostat, electromagnetic analysis, vacuum system, power handling and conversion, NBI, RF heating device, tritium system, neutronics, maintenance, cooling system and layout of facilities. The engineering comparison of a divertor with pumped limiters and safety analysis of reactor systems are also conducted. (author)

  10. Conceptual design of fusion experimental reactor (FER)

    International Nuclear Information System (INIS)

    1985-01-01

    The Fusion Experimental Reactor (FER) being developed at JAERI as a next generation tokamak to JT-60 has a major mission of realizing a self-ignited long-burning DT plasma and demonstrating engineering feasibility. During FY82 and FY83 a comprehensive and intensive conceptual design study has been conducted for a pulsed operation FER as a reference option which employs a conventional inductive current drive and a double-null divertor. In parallel with the reference design, studies have been carried out to evaluate advanced reactor concepts such as quasi-steady state operation and steady state operation based on RF current drive and pumped limiter, and comparative studies for single-null divertor/pumped limiter. This report presents major results obtained primarily from FY83 design studies, while the results of FY82 design studies are described in previous references (JAERI-M 83-213--216). (author)

  11. Premature activation of the paramyxovirus fusion protein before target cell attachment with corruption of the viral fusion machinery.

    Science.gov (United States)

    Farzan, Shohreh F; Palermo, Laura M; Yokoyama, Christine C; Orefice, Gianmarco; Fornabaio, Micaela; Sarkar, Aurijit; Kellogg, Glen E; Greengard, Olga; Porotto, Matteo; Moscona, Anne

    2011-11-04

    Paramyxoviruses, including the childhood pathogen human parainfluenza virus type 3, enter host cells by fusion of the viral and target cell membranes. This fusion results from the concerted action of its two envelope glycoproteins, the hemagglutinin-neuraminidase (HN) and the fusion protein (F). The receptor-bound HN triggers F to undergo conformational changes that render it competent to mediate fusion of the viral and cellular membranes. We proposed that, if the fusion process could be activated prematurely before the virion reaches the target host cell, infection could be prevented. We identified a small molecule that inhibits paramyxovirus entry into target cells and prevents infection. We show here that this compound works by an interaction with HN that results in F-activation prior to receptor binding. The fusion process is thereby prematurely activated, preventing fusion of the viral membrane with target cells and precluding viral entry. This first evidence that activation of a paramyxovirus F can be specifically induced before the virus contacts its target cell suggests a new strategy with broad implications for the design of antiviral agents.

  12. Rho GTPase activity modulates paramyxovirus fusion protein-mediated cell-cell fusion

    International Nuclear Information System (INIS)

    Schowalter, Rachel M.; Wurth, Mark A.; Aguilar, Hector C.; Lee, Benhur; Moncman, Carole L.; McCann, Richard O.; Dutch, Rebecca Ellis

    2006-01-01

    The paramyxovirus fusion protein (F) promotes fusion of the viral envelope with the plasma membrane of target cells as well as cell-cell fusion. The plasma membrane is closely associated with the actin cytoskeleton, but the role of actin dynamics in paramyxovirus F-mediated membrane fusion is unclear. We examined cell-cell fusion promoted by two different paramyxovirus F proteins in three cell types in the presence of constitutively active Rho family GTPases, major cellular coordinators of actin dynamics. Reporter gene and syncytia assays demonstrated that expression of either Rac1 V12 or Cdc42 V12 could increase cell-cell fusion promoted by the Hendra or SV5 glycoproteins, though the effect was dependent on the cell type expressing the viral glycoproteins. In contrast, RhoA L63 decreased cell-cell fusion promoted by Hendra glycoproteins but had little affect on SV5 F-mediated fusion. Also, data suggested that GTPase activation in the viral glycoprotein-containing cell was primarily responsible for changes in fusion. Additionally, we found that activated Cdc42 promoted nuclear rearrangement in syncytia

  13. The dengue virus type 2 envelope protein fusion peptide is essential for membrane fusion

    International Nuclear Information System (INIS)

    Huang, Claire Y.-H.; Butrapet, Siritorn; Moss, Kelly J.; Childers, Thomas; Erb, Steven M.; Calvert, Amanda E.; Silengo, Shawn J.; Kinney, Richard M.; Blair, Carol D.; Roehrig, John T.

    2010-01-01

    The flaviviral envelope (E) protein directs virus-mediated membrane fusion. To investigate membrane fusion as a requirement for virus growth, we introduced 27 unique mutations into the fusion peptide of an infectious cDNA clone of dengue 2 virus and recovered seven stable mutant viruses. The fusion efficiency of the mutants was impaired, demonstrating for the first time the requirement for specific FP AAs in optimal fusion. Mutant viruses exhibited different growth kinetics and/or genetic stabilities in different cell types and adult mosquitoes. Virus particles could be recovered following RNA transfection of cells with four lethal mutants; however, recovered viruses could not re-infect cells. These viruses could enter cells, but internalized virus appeared to be retained in endosomal compartments of infected cells, thus suggesting a fusion blockade. Mutations of the FP also resulted in reduced virus reactivity with flavivirus group-reactive antibodies, confirming earlier reports using virus-like particles.

  14. Impact of fluorescent protein fusions on the bacterial flagellar motor.

    Science.gov (United States)

    Heo, M; Nord, A L; Chamousset, D; van Rijn, E; Beaumont, H J E; Pedaci, F

    2017-10-03

    Fluorescent fusion proteins open a direct and unique window onto protein function. However, they also introduce the risk of perturbation of the function of the native protein. Successful applications of fluorescent fusions therefore rely on a careful assessment and minimization of the side effects, but such insight is still lacking for many applications. This is particularly relevant in the study of the internal dynamics of motor proteins, where both the chemical and mechanical reaction coordinates can be affected. Fluorescent proteins fused to the stator of the Bacterial Flagellar Motor (BFM) have previously been used to unveil the motor subunit dynamics. Here we report the effects on single motors of three fluorescent proteins fused to the stators, all of which altered BFM behavior. The torque generated by individual stators was reduced while their stoichiometry remained unaffected. MotB fusions decreased the switching frequency and induced a novel bias-dependent asymmetry in the speed in the two directions. These effects could be mitigated by inserting a linker at the fusion point. These findings provide a quantitative account of the effects of fluorescent fusions to the stator on BFM dynamics and their alleviation- new insights that advance the use of fluorescent fusions to probe the dynamics of protein complexes.

  15. Conceptual design of fusion experimental reactor (FER)

    International Nuclear Information System (INIS)

    1984-01-01

    Conceptual Design of Fusion Experimental Reactor (FER) of which the objective will be to realize self-ignition with D-T reaction is reported. Mechanical Configurations of FER are characterized with a noncircular plasma and a double-null divertor. The primary aim of design studies is to demonstrate fissibility of reactor structures as compact and simple as possible with removable torus sectors. The structures of each component such as a first-wall, blanket, shielding, divertor, magnet and so on have been designed. It is also discussed about essential reactor plant system requirements. In addition to the above, a brief concept of a steady-state reactor based on RF current drive is also discussed. The main aim, in this time, is to examine physical studies of a possible RF steady-state reactor. (author)

  16. Recombinant fusion protein of albumin-retinol binding protein inactivates stellate cells

    Energy Technology Data Exchange (ETDEWEB)

    Choi, Soyoung; Park, Sangeun; Kim, Suhyun [Laboratory of Cellular Oncology, Korea University Graduate School of Medicine, Ansan, Gyeonggi do 425-707 (Korea, Republic of); Lim, Chaeseung [Department of Laboratory Medicine, Korea University Guro Hospital, Seoul 152-703 (Korea, Republic of); Kim, Jungho [Department of Life Science, Sogang University, Seoul 121-742 (Korea, Republic of); Cha, Dae Ryong [Department of Internal Medicine, Korea University Ansan Hospital, Ansan, Gyeonggi do 425-020 (Korea, Republic of); Oh, Junseo, E-mail: ohjs@korea.ac.kr [Laboratory of Cellular Oncology, Korea University Graduate School of Medicine, Ansan, Gyeonggi do 425-707 (Korea, Republic of)

    2012-02-03

    Highlights: Black-Right-Pointing-Pointer We designed novel recombinant albumin-RBP fusion proteins. Black-Right-Pointing-Pointer Expression of fusion proteins inactivates pancreatic stellate cells (PSCs). Black-Right-Pointing-Pointer Fusion proteins are successfully internalized into and inactivate PSCs. Black-Right-Pointing-Pointer RBP moiety mediates cell specific uptake of fusion protein. -- Abstract: Quiescent pancreatic- (PSCs) and hepatic- (HSCs) stellate cells store vitamin A (retinol) in lipid droplets via retinol binding protein (RBP) receptor and, when activated by profibrogenic stimuli, they transform into myofibroblast-like cells which play a key role in the fibrogenesis. Despite extensive investigations, there is, however, currently no appropriate therapy available for tissue fibrosis. We previously showed that the expression of albumin, composed of three homologous domains (I-III), inhibits stellate cell activation, which requires its high-affinity fatty acid-binding sites asymmetrically distributed in domain I and III. To attain stellate cell-specific uptake, albumin (domain I/III) was coupled to RBP; RBP-albumin{sup domain} {sup III} (R-III) and albumin{sup domain} {sup I}-RBP-albumin{sup III} (I-R-III). To assess the biological activity of fusion proteins, cultured PSCs were used. Like wild type albumin, expression of R-III or I-R-III in PSCs after passage 2 (activated PSCs) induced phenotypic reversal from activated to fat-storing cells. On the other hand, R-III and I-R-III, but not albumin, secreted from transfected 293 cells were successfully internalized into and inactivated PSCs. FPLC-purified R-III was found to be internalized into PSCs via caveolae-mediated endocytosis, and its efficient cellular uptake was also observed in HSCs and podocytes among several cell lines tested. Moreover, tissue distribution of intravenously injected R-III was closely similar to that of RBP. Therefore, our data suggest that albumin-RBP fusion protein comprises

  17. Protein fold recognition using geometric kernel data fusion.

    Science.gov (United States)

    Zakeri, Pooya; Jeuris, Ben; Vandebril, Raf; Moreau, Yves

    2014-07-01

    Various approaches based on features extracted from protein sequences and often machine learning methods have been used in the prediction of protein folds. Finding an efficient technique for integrating these different protein features has received increasing attention. In particular, kernel methods are an interesting class of techniques for integrating heterogeneous data. Various methods have been proposed to fuse multiple kernels. Most techniques for multiple kernel learning focus on learning a convex linear combination of base kernels. In addition to the limitation of linear combinations, working with such approaches could cause a loss of potentially useful information. We design several techniques to combine kernel matrices by taking more involved, geometry inspired means of these matrices instead of convex linear combinations. We consider various sequence-based protein features including information extracted directly from position-specific scoring matrices and local sequence alignment. We evaluate our methods for classification on the SCOP PDB-40D benchmark dataset for protein fold recognition. The best overall accuracy on the protein fold recognition test set obtained by our methods is ∼ 86.7%. This is an improvement over the results of the best existing approach. Moreover, our computational model has been developed by incorporating the functional domain composition of proteins through a hybridization model. It is observed that by using our proposed hybridization model, the protein fold recognition accuracy is further improved to 89.30%. Furthermore, we investigate the performance of our approach on the protein remote homology detection problem by fusing multiple string kernels. The MATLAB code used for our proposed geometric kernel fusion frameworks are publicly available at http://people.cs.kuleuven.be/∼raf.vandebril/homepage/software/geomean.php?menu=5/. © The Author 2014. Published by Oxford University Press.

  18. Ceramic sphere-pac breeder design for fusion blankets

    International Nuclear Information System (INIS)

    Gierszewski, P.J.; Sullivan, J.D.

    1991-01-01

    Randomly packed beds of ceramic spheres are a practical approach to surrounding fusion plasmas with tritium-breeding material. This paper examines the general properties of sphere-pac beds for application in fusion breeder blankets. The design considerations and models are reviewed for packing, tritium breeding and recovery, thermal conductivity, purge-gas pressure drop, mechanical behavior and fabrication. The design correlations are compared against available fusion ceramic data. Specific conclusions are that ternary (three-size) beds are not attractive for fusion blankets, and that the fusion spheres should be as large as possible subject primarily to packing constraints. (orig.)

  19. Escherichia coli fusion carrier proteins act as solubilizing agents for recombinant uncoupling protein 1 through interactions with GroEL

    International Nuclear Information System (INIS)

    Douette, Pierre; Navet, Rachel; Gerkens, Pascal; Galleni, Moreno; Levy, Daniel; Sluse, Francis E.

    2005-01-01

    Fusing recombinant proteins to highly soluble partners is frequently used to prevent aggregation of recombinant proteins in Escherichia coli. Moreover, co-overexpression of prokaryotic chaperones can increase the amount of properly folded recombinant proteins. To understand the solubility enhancement of fusion proteins, we designed two recombinant proteins composed of uncoupling protein 1 (UCP1), a mitochondrial membrane protein, in fusion with MBP or NusA. We were able to express soluble forms of MBP-UCP1 and NusA-UCP1 despite the high hydrophobicity of UCP1. Furthermore, the yield of soluble fusion proteins depended on co-overexpression of GroEL that catalyzes folding of polypeptides. MBP-UCP1 was expressed in the form of a non-covalent complex with GroEL. MBP-UCP1/GroEL was purified and characterized by dynamic light scattering, gel filtration, and electron microscopy. Our findings suggest that MBP and NusA act as solubilizing agents by forcing the recombinant protein to pass through the bacterial chaperone pathway in the context of fusion protein

  20. The Multifaceted Role of SNARE Proteins in Membrane Fusion.

    Science.gov (United States)

    Han, Jing; Pluhackova, Kristyna; Böckmann, Rainer A

    2017-01-01

    Membrane fusion is a key process in all living organisms that contributes to a variety of biological processes including viral infection, cell fertilization, as well as intracellular transport, and neurotransmitter release. In particular, the various membrane-enclosed compartments in eukaryotic cells need to exchange their contents and communicate across membranes. Efficient and controllable fusion of biological membranes is known to be driven by cooperative action of SNARE proteins, which constitute the central components of the eukaryotic fusion machinery responsible for fusion of synaptic vesicles with the plasma membrane. During exocytosis, vesicle-associated v-SNARE (synaptobrevin) and target cell-associated t-SNAREs (syntaxin and SNAP-25) assemble into a core trans-SNARE complex. This complex plays a versatile role at various stages of exocytosis ranging from the priming to fusion pore formation and expansion, finally resulting in the release or exchange of the vesicle content. This review summarizes current knowledge on the intricate molecular mechanisms underlying exocytosis triggered and catalyzed by SNARE proteins. Particular attention is given to the function of the peptidic SNARE membrane anchors and the role of SNARE-lipid interactions in fusion. Moreover, the regulatory mechanisms by synaptic auxiliary proteins in SNARE-driven membrane fusion are briefly outlined.

  1. Target design for heavy ion beam fusion

    International Nuclear Information System (INIS)

    Meyer-ter-Vehn, J.; Metzler, N.

    1981-07-01

    Target design for Heavy Ion Beam Fusion and related physics are discussed. First, a modified version of the Kidder-Bodner model for pellet gain is presented and is used to define the working point (Esub(beam) = 4.8 MJ, Gain 83) for a reactor size target. Secondly, stopping of heavy ions in hot dense plasma is investigated and numerical results for stopping powers and ranges of 10 GeV Bi-ions in Pb, Li, and PbLi-alloy are given. Finally, results of an explicit implosion calculation, using the 1-D code MINIHY, are discussed in detail. The hydrodynamic efficiency is found to be about 5%. Special attention is given to the shock sequence leading to the ignition configuration. Also the growth of Rayleigh-Taylor instability at the absorber-pusher interface is estimated. (orig.)

  2. The yeast cell fusion protein Prm1p requires covalent dimerization to promote membrane fusion.

    Directory of Open Access Journals (Sweden)

    Alex Engel

    2010-05-01

    Full Text Available Prm1p is a multipass membrane protein that promotes plasma membrane fusion during yeast mating. The mechanism by which Prm1p and other putative regulators of developmentally controlled cell-cell fusion events facilitate membrane fusion has remained largely elusive. Here, we report that Prm1p forms covalently linked homodimers. Covalent Prm1p dimer formation occurs via intermolecular disulfide bonds of two cysteines, Cys-120 and Cys-545. PRM1 mutants in which these cysteines have been substituted are fusion defective. These PRM1 mutants are normally expressed, retain homotypic interaction and can traffic to the fusion zone. Because prm1-C120S and prm1-C545S mutants can form covalent dimers when coexpressed with wild-type PRM1, an intermolecular C120-C545 disulfide linkage is inferred. Cys-120 is adjacent to a highly conserved hydrophobic domain. Mutation of a charged residue within this hydrophobic domain abrogates formation of covalent dimers, trafficking to the fusion zone, and fusion-promoting activity. The importance of intermolecular disulfide bonding informs models regarding the mechanism of Prm1-mediated cell-cell fusion.

  3. Fusion reactor design studies: standard accounts for cost estimates

    International Nuclear Information System (INIS)

    Schulte, S.C.; Willke, T.L.; Young, J.R.

    1978-05-01

    The fusion reactor design studies--standard accounts for cost estimates provides a common format from which to assess the economic character of magnetically confined fusion reactor design concepts. The format will aid designers in the preparation of design concept costs estimates and also provide policymakers with a tool to assist in appraising which design concept may be economically promising. The format sets forth a categorization and accounting procedure to be used when estimating fusion reactor busbar energy cost that can be easily and consistently applied. Reasons for developing the procedure, explanations of the procedure, justifications for assumptions made in the procedure, and the applicability of the procedure are described in this document. Adherence to the format when evaluating prospective fusion reactor design concepts will result in the identification of the more promising design concepts thus enabling the fusion power alternatives with better economic potential to be quickly and efficiently developed

  4. Neutronics issues in fusion-fission hybrid reactor design

    International Nuclear Information System (INIS)

    Liu Chengan

    1995-01-01

    The coupled neutron and γ-ray transport equations and nuclear number density equations, and its computer program systems concerned in fusion-fission hybrid reactor design are briefly described. The current status and focal point for coming work of nuclear data used in fusion reactor design are explained

  5. Recent designs for advanced fusion reactor blankets

    International Nuclear Information System (INIS)

    Sze, D.K.

    1994-01-01

    A series of reactor design studies based on the Tokamak configuration have been carried out under the direction of Professor Robert Conn of UCLA. They are called ARIES-I through IV. The key mission of these studies is to evaluate the attractiveness of fusion assuming different degrees of advancement in either physics or engineering development. This paper discusses the directions and conclusions of the blanket and related engineering systems for those design studies. ARIES-1 investigated the use of SiC composite as the structural material to increase the blanket temperature and reduce the blanket activation. Li 2 ZrO 3 was used as the breeding material due to its high temperature stability and good tritium recovery characteristics. The ARIES-IV is a modification of ARIES-1. The plasma was in the second stability regime. Li 2 O was used as the breeding material to remove Zr. A gaseous divertor was used to replace the conventional divertor so that high Z divertor target is not required. The physics of ARIES-II was the same as ARIES-IV. The engineering design of the ARIES-II was based on a self-cooled lithium blanket with a V-alloy as the structural material. Even though it was assumed that the plasma was in the second stability regime, the plasma beta was still rather low (3.4%). The ARIES-III is an advanced fuel (D- 3 He) tokamak reactor. The reactor design assumed major advancement on the physics, with a plasma beta of 23.9%. A conventional structural material is acceptable due to the low neutron wall loading. From the radiation damage point of view, the first wall can last the life of the reactor, which is expected to be a major advantage from the engineering design and waste disposal point of view

  6. Osteoinductive recombinant silk fusion proteins for bone regeneration.

    Science.gov (United States)

    Dinjaski, Nina; Plowright, Robyn; Zhou, Shun; Belton, David J; Perry, Carole C; Kaplan, David L

    2017-02-01

    Protein polymers provide a unique opportunity for tunable designs of material systems due to the genetic basis of sequence control. To address the challenge of biomineralization interfaces with protein based materials, we genetically engineered spider silks to design organic-inorganic hybrid systems. The spider silk inspired domain (SGRGGLGGQG AGAAAAAGGA GQGGYGGLGSQGT) 15 served as an organic scaffold to control material stability and to allow multiple modes of processing, whereas the hydroxyapatite binding domain VTKHLNQISQSY (VTK), provided control over osteogenesis. The VTK domain was fused either to the N-, C- or both terminals of the spider silk domain to understand the effect of position on material properties and mineralization. The addition of the VTK domain to silk did not affect the physical properties of the silk recombinant constructs, but it had a critical role in the induction of biomineralization. When the VTK domain was placed on both the C- and N-termini the formation of crystalline hydroxyapatite was significantly increased. In addition, all of the recombinant proteins in film format supported the growth and proliferation of human mesenchymal stem cells (hMSCs). Importantly, the presence of the VTK domain enhanced osteoinductive properties up to 3-fold compared to the control (silk alone without VTK). Therefore, silk-VTK fusion proteins have been shown suitable for mineralization and functionalization for specific biomedical applications. Organic-inorganic interfaces are integral to biomaterial functions in many areas of repair and regeneration. Several protein polymers have been investigated for this purpose. Despite their success the limited options to fine-tune their material properties, degradation patterns and functionalize them for each specific biomedical application limits their application. Various studies have shown that the biological performance of such proteins can be improved by genetic engineering. The present study provides data

  7. Conceptual design of imploding liner fusion reactors

    International Nuclear Information System (INIS)

    Turchi, P.J.; Robson, A.E.

    1976-01-01

    The basic new ingredient is the concept of rotationally stabilized liquid metal liners accelerated with free pistons. The liner motion is constrained on its outer surface by the pistons, laterally by channel walls, during acceleration, and on its inner surface, where megagauss field levels are attained by the centrifugal motion of the liner material. In this way, stable, reversible motion of the liner should be possible, permitting repetitive, pulsed operation at interior pressures far greater than can be allowed in static conductor systems. Such higher operating pressures permit the use of simple plasma geometries, such as theta pinches, with greatly reduced dimensions. Furthermore, the implosion of thick, lithium-bearing liners with large radial compression ratios inherently provides the plasma with a surrounding blanket of neutron absorbing liquid metal, thereby substantially reducing the problems of induced radioactivity and first wall damage that haunt conventional fusion reactor designs. The following article discusses the basic operation of liner reactors and several important features influencing their design

  8. Fusion reactor blanket-main design aspects

    International Nuclear Information System (INIS)

    Strebkov, Yu.; Sidorov, A.; Danilov, I.

    1994-01-01

    The main function of the fusion reactor blanket is ensuring tritium breeding and radiation shield. The blanket version depends on the reactor type (experimental, DEMO, commercial) and its parameters. Blanket operation conditions are defined with the heat flux, neutron load/fluence, cyclic operation, dynamic heating/force loading, MHD effects etc. DEMO/commercial blanket design is distinguished e.g. by rather high heat load and neutron fluence - up to 100 W/cm 2 and 7 MWa/m 2 accordingly. This conditions impose specific requirements for the materials, structure, maintenance of the blanket and its most loaded components - FW and limiter. The liquid Li-Pb eutectic is one of the possible breeder for different kinds of blanket in view of its advantages one of which is the blanket convertibility that allow to have shielding blanket (borated water) or breeding one (Li-Pb eutectic). Using Li-Pb eutectic for both ITER and DEMO blankets have been considered. In the conceptual ITER design the solid eutectic blanket was carried out. The liquid eutectic breeder/coolant is suggested also for the advanced (high parameter) blanket

  9. Conceptual design of inertial confinement fusion power plant

    International Nuclear Information System (INIS)

    Mima, Kunioki; Yamanaka, Tatsuhiko; Nakai, Sadao

    1994-01-01

    Presented is the status of the conceptual design studies of inertial confinement fusion reactors. The recent achievements of the laser fusion research enable us to refine the conceptual design of the power plant. In the paper, main features of several new conceptual designs of ICF reactor; KOYO, SIRIUS-P, HYLIFE-II and so on are summarized. In particular, the target design and the reactor chamber design are described. Finally, the overview of the laser fusion reactor and the irradiation system is also described. (author)

  10. Computational and biological characterization of fusion proteins of two insecticidal proteins for control of insect pests.

    Science.gov (United States)

    Javaid, Shaista; Naz, Sehrish; Amin, Imran; Jander, Georg; Ul-Haq, Zaheer; Mansoor, Shahid

    2018-03-19

    Sucking pests pose a serious agricultural challenge, as available transgenic technologies such as Bacillus thuringiensis crystal toxins (Bt) are not effective against them. One approach is to produce fusion protein toxins for the control of these pests. Two protein toxins, Hvt (ω-atracotoxin from Hadronyche versuta) and onion leaf lectin, were translationally fused to evaluate the negative effects of fusion proteins on Phenacoccus solenopsis (mealybug), a phloem-feeding insect pest. Hvt was cloned both N-terminally (HL) and then C-terminally (LH) in the fusion protein constructs, which were expressed transiently in Nicotiana tabacum using a Potato Virus X (PVX) vector. The HL fusion protein was found to be more effective against P. solenopsis, with an 83% mortality rate, as compared to the LH protein, which caused 65% mortality. Hvt and lectin alone caused 42% and 45%, respectively, under the same conditions. Computational studies of both fusion proteins showed that the HL protein is more stable than the LH protein. Together, these results demonstrate that translational fusion of two insecticidal proteins improved the insecticidal activity relative to each protein individually and could be expressed in transgenic plants for effective control of sucking pests.

  11. Study on Fusion Protein and Its gene in Baculovirus Specificity

    International Nuclear Information System (INIS)

    Nemr, W.A.H.

    2012-01-01

    Baculoviruses are subdivided into two groups depending on the type of budded virus envelop fusion protein; group I utilized gp64 which include the most of nucleopolyhedroviruses (NPVs), group II utilized F protein which include the remnants of NPVs and all Granuloviruses (GVs). Recent studies reported the viral F protein coding gene as a host cellular sourced gene and may evolutionary acquired from the host genome referring to phylogeny analysis of fusion proteins. Thus, it was deduced that F protein coding gene is species- specific nucleotide sequence related to the type of the specific host and if virus could infect an unexpected host, the resulted virus may encode a vary F gene. In this regard, the present study utilized the mentioned properties of F gene in an attempt to produce a model of specific and more economic wider range granulovirus bio- pesticide able to infect both Spodoptera littoralis and Phthorimaea operculella larvae. Multiple sequence alignment and phylogeny analysis were performed on six members of group II baculovirus, novel universal PCR primers were manually designed from the conserved regions in the alignment graph, targeted to amplify species- specific sequence entire F gene open reading frame (ORF) which is useful in molecular identification of baculovirus in unknown samples. So, the PCR product of SpliGV used to prepare a specific probe for the F gene of this type of virus. Results reflected that it is possible to infect S. littoralis larvae by PhopGV if injected into larval haemocoel, the resulted virus of this infection showed by using DNA hybridization technique to be encode to F gene homologous with the F gene of Spli GV, which is revealed that the resulted virus acquired this F gene sequence from the host genome after infection. Consequently, these results may infer that if genetic aberrations occur in the host genome, this may affect in baculoviral infectivity. So, this study aimed to investigate the effect of gamma radiation at

  12. Fusion reactor design and technology 1986. V. 1

    International Nuclear Information System (INIS)

    1987-01-01

    The first volume of the Proceedings of the Fourth Technical Committee Meeting and Workshop on Fusion Reactor Design and Technology organized by the IAEA (Yalta, 26 May - 6 June 1986) includes 36 papers devoted to the following topics: fusion programmes (3 papers), tokamaks (15 papers), non-tokamak reactors and open systems (9 papers), inertial confinement concepts (5 papers), fission-fusion hybrids (4 papers). Each of these papers has a separate abstract. Refs, figs and tabs

  13. Review of mirror fusion reactor designs

    International Nuclear Information System (INIS)

    Bender, D.J.

    1977-01-01

    Three magnetic confinement concepts, based on the mirror principle, are described. These mirror concepts are summarized as follows: (1) fusion-fission hybrid reactor, (2) tandem mirror reactor, and (3) reversed field mirror reactor

  14. Radiological design criteria for fusion power test facilities

    International Nuclear Information System (INIS)

    Singh, M.S.; Campbell, G.W.

    1982-01-01

    The quest for fusion power and understanding of plasma physics has resulted in planning, design, and construction of several major fusion power test facilities, based largely on magnetic and inertial confinement concepts. We have considered radiological design aspects of the Joint European Torus (JET), Livermore Mirror and Inertial Fusion projects, and Princeton Tokamak. Our analyses on radiological design criteria cover acceptable exposure levels at the site boundary, man-rem doses for plant personnel and population at large, based upon experience gained for the fission reactors, and on considerations of cost-benefit analyses

  15. Fusion reactor design: On the road to commercialization

    International Nuclear Information System (INIS)

    Kulcinski, G.L.

    1984-01-01

    The worldwide effort in fusion is now approximately 2 billion dollars per year and over 12 billion dollars has been invested since 1951 in developing this energy source for the 21st century. A vital component of the past efforts in fusion research has been the conceptual design activities performed by scientists and engineers around the world. Almost 80 such major designs of Tokamak, Mirror, Laser and Ion Beam Reactors have been published and this article discusses how recent conceptual designs have afftected our perception of future fusion reactor performance. (orig.) [de

  16. Initial trade and design studies for the fusion engineering device

    International Nuclear Information System (INIS)

    Flanagan, C.A.; Steiner, D.; Smith, G.E.

    1981-06-01

    The Magnetic Fusion Energy Engineering Act of 1980 calls for the operation of a Fusion Engineering Device (FED) by 1990. It is the intent of the Act that the FED, in combination with other testing facilities, will establish the engineering feasibility of magnetic fusion energy. The Fusion Engineering Design Center (FEDC), under the guidance of a Technical Management Board (TMB), initiated a program of trade and design studies in October 1980 to support the selection of the FED concept. This document presents the results of these initial trade and design studies. Based on these results, a baseline configuration has been identified and the Design Center effort for the remainder of the fiscal year will be devoted to the development of a self-consistent FED design description

  17. Safety considerations in next step fusion design and beyond

    International Nuclear Information System (INIS)

    Holland, D.F.

    1990-01-01

    Recent U.S. and international design studies provide insights into the potential safety and environmental advantages of fusion as well as the development needed to realize this potential. We in the Fusion Safety Program at EG ampersand G Idaho have analyzed the Compact Ignition Tokamak (CIT), the International Thermonuclear Engineering Reactor (ITER), and the Advanced Reactor Innovative Engineering Study (ARIES). I have reviewed these three designs to determine issues related to meeting the safety and the environmental goals that guide fusion development in the U.S. The paper lists safety and environmental issues that are generic to fusion and approaches to favorably resolve each issue. The technical developments that have the highest potential of contributing to improving the safety and environmental attractiveness of fusion are identified and discussed. These developments are in the areas of low-activation materials, plasma- facing components, and plasma physics relating to off-normal plasma events and tritium burn-up. 8 refs., 7 tabs

  18. Conceptual design of light ion beam inertia nuclear fusion reactors

    International Nuclear Information System (INIS)

    1983-07-01

    Light ion beam, inertia nuclear fusion system drew attention recently as one of the nuclear fusion systems for power reactors in the history of the research on nuclear fusion. Its beginning seemed to be the judgement that the implosion of fusion fuel pellets with light ions can be realized with the light ions which can be obtained in view of accelerator techniques. Of course, in order to generate practically usable nuclear fusion reaction by this system and maintain it, many technical difficulties must be overcome. This research was carried out for the purpose of discovering such technical problems and searching for their solution. At the time of doing the works, the following policy was adopted. Though their is the difference of fine and rough, the design of a whole reactor system is performed conformably. In order to make comparison with other reactor types and nuclear fusion systems, the design is carried out as the power plant of about one million kWe output. As the extent of the design, the works at conceptual design stage are performed to present the concept of design which satisfies the required function. Basically, the design is made from conservative standpoint. This research of design was started in 1981, and in fiscal 1982, the mutual adjustment among the design of respective parts was performed on the basis of the results in 1981, and the possible revision and new proposal were investigated. (Kako, I.)

  19. Regulation of Exocytotic Fusion Pores by SNARE Protein Transmembrane Domains

    Directory of Open Access Journals (Sweden)

    Zhenyong Wu

    2017-10-01

    Full Text Available Calcium-triggered exocytotic release of neurotransmitters and hormones from neurons and neuroendocrine cells underlies neuronal communication, motor activity and endocrine functions. The core of the neuronal exocytotic machinery is composed of soluble N-ethyl maleimide sensitive factor attachment protein receptors (SNAREs. Formation of complexes between vesicle-attached v- and plasma-membrane anchored t-SNAREs in a highly regulated fashion brings the membranes into close apposition. Small, soluble proteins called Complexins (Cpx and calcium-sensing Synaptotagmins cooperate to block fusion at low resting calcium concentrations, but trigger release upon calcium increase. A growing body of evidence suggests that the transmembrane domains (TMDs of SNARE proteins play important roles in regulating the processes of fusion and release, but the mechanisms involved are only starting to be uncovered. Here we review recent evidence that SNARE TMDs exert influence by regulating the dynamics of the fusion pore, the initial aqueous connection between the vesicular lumen and the extracellular space. Even after the fusion pore is established, hormone release by neuroendocrine cells is tightly controlled, and the same may be true of neurotransmitter release by neurons. The dynamics of the fusion pore can regulate the kinetics of cargo release and the net amount released, and can determine the mode of vesicle recycling. Manipulations of SNARE TMDs were found to affect fusion pore properties profoundly, both during exocytosis and in biochemical reconstitutions. To explain these effects, TMD flexibility, and interactions among TMDs or between TMDs and lipids have been invoked. Exocytosis has provided the best setting in which to unravel the underlying mechanisms, being unique among membrane fusion reactions in that single fusion pores can be probed using high-resolution methods. An important role will likely be played by methods that can probe single fusion pores

  20. Localized cyclic AMP-dependent protein kinase activity is required for myogenic cell fusion

    International Nuclear Information System (INIS)

    Mukai, Atsushi; Hashimoto, Naohiro

    2008-01-01

    Multinucleated myotubes are formed by fusion of mononucleated myogenic progenitor cells (myoblasts) during terminal skeletal muscle differentiation. In addition, myoblasts fuse with myotubes, but terminally differentiated myotubes have not been shown to fuse with each other. We show here that an adenylate cyclase activator, forskolin, and other reagents that elevate intracellular cyclic AMP (cAMP) levels induced cell fusion between small bipolar myotubes in vitro. Then an extra-large myotube, designated a 'myosheet,' was produced by both primary and established mouse myogenic cells. Myotube-to-myotube fusion always occurred between the leading edge of lamellipodia at the polar end of one myotube and the lateral plasma membrane of the other. Forskolin enhanced the formation of lamellipodia where cAMP-dependent protein kinase (PKA) was accumulated. Blocking enzymatic activity or anchoring of PKA suppressed forskolin-enhanced lamellipodium formation and prevented fusion of multinucleated myotubes. Localized PKA activity was also required for fusion of mononucleated myoblasts. The present results suggest that localized PKA plays a pivotal role in the early steps of myogenic cell fusion, such as cell-to-cell contact/recognition through lamellipodium formation. Furthermore, the localized cAMP-PKA pathway might be involved in the specification of the fusion-competent areas of the plasma membrane in lamellipodia of myogenic cells

  1. Viral membrane fusion: is glycoprotein G of rhabdoviruses a representative of a new class of viral fusion proteins?

    Directory of Open Access Journals (Sweden)

    A.T. Da Poian

    2005-06-01

    Full Text Available Enveloped viruses always gain entry into the cytoplasm by fusion of their lipid envelope with a cell membrane. Some enveloped viruses fuse directly with the host cell plasma membrane after virus binding to the cell receptor. Other enveloped viruses enter the cells by the endocytic pathway, and fusion depends on the acidification of the endosomal compartment. In both cases, virus-induced membrane fusion is triggered by conformational changes in viral envelope glycoproteins. Two different classes of viral fusion proteins have been described on the basis of their molecular architecture. Several structural data permitted the elucidation of the mechanisms of membrane fusion mediated by class I and class II fusion proteins. In this article, we review a number of results obtained by our laboratory and by others that suggest that the mechanisms involved in rhabdovirus fusion are different from those used by the two well-studied classes of viral glycoproteins. We focus our discussion on the electrostatic nature of virus binding and interaction with membranes, especially through phosphatidylserine, and on the reversibility of the conformational changes of the rhabdovirus glycoprotein involved in fusion. Taken together, these data suggest the existence of a third class of fusion proteins and support the idea that new insights should emerge from studies of membrane fusion mediated by the G protein of rhabdoviruses. In particular, the elucidation of the three-dimensional structure of the G protein or even of the fusion peptide at different pH's might provide valuable information for understanding the fusion mechanism of this new class of fusion proteins.

  2. Conceptual design study of Fusion Experimental Reactor (FY87FER)

    International Nuclear Information System (INIS)

    1988-05-01

    The design study of Fusion Experimental Reactor(FER) which has been proposed to be the next step fusion device has been conducted by JAERI Reactor System Laboratory since 1982 and by FER design team since 1984. This is the final report of the FER design team program and describes the results obtained in FY1987 (partially in FY1986) activities. The contents of this report consist of the reference design which is based on the guideline in FY1986 by the Subcomitees set up in Nuclear Fusion Council of Atomic Energy Commission of Japan, the Low-Physics-Risk reactor design for achieving physics mission more reliably and the system study of FER design candidates including above two designs. (author)

  3. Comparative immunoblot analysis with 10 different, partially overlapping recombinant fusion proteins derived from 5 different cytomegalovirus proteins

    NARCIS (Netherlands)

    van Zanten, J.; LAZZAROTTO, T; CAMPISI, B; VORNHAGEN, R; JAHN, G; LANDINI, MP; The, T. Hauw

    Ten fusion proteins derived from five various CMV encoded proteins were used for the detection of specific antibody response by immunoblot technique in sera from renal transplant recipients. The fusion proteins were derived from the following CMV specific proteins: the assembly protein ppUL80a with

  4. An Approach to Automated Fusion System Design and Adaptation

    Directory of Open Access Journals (Sweden)

    Alexander Fritze

    2017-03-01

    Full Text Available Industrial applications are in transition towards modular and flexible architectures that are capable of self-configuration and -optimisation. This is due to the demand of mass customisation and the increasing complexity of industrial systems. The conversion to modular systems is related to challenges in all disciplines. Consequently, diverse tasks such as information processing, extensive networking, or system monitoring using sensor and information fusion systems need to be reconsidered. The focus of this contribution is on distributed sensor and information fusion systems for system monitoring, which must reflect the increasing flexibility of fusion systems. This contribution thus proposes an approach, which relies on a network of self-descriptive intelligent sensor nodes, for the automatic design and update of sensor and information fusion systems. This article encompasses the fusion system configuration and adaptation as well as communication aspects. Manual interaction with the flexibly changing system is reduced to a minimum.

  5. An Approach to Automated Fusion System Design and Adaptation.

    Science.gov (United States)

    Fritze, Alexander; Mönks, Uwe; Holst, Christoph-Alexander; Lohweg, Volker

    2017-03-16

    Industrial applications are in transition towards modular and flexible architectures that are capable of self-configuration and -optimisation. This is due to the demand of mass customisation and the increasing complexity of industrial systems. The conversion to modular systems is related to challenges in all disciplines. Consequently, diverse tasks such as information processing, extensive networking, or system monitoring using sensor and information fusion systems need to be reconsidered. The focus of this contribution is on distributed sensor and information fusion systems for system monitoring, which must reflect the increasing flexibility of fusion systems. This contribution thus proposes an approach, which relies on a network of self-descriptive intelligent sensor nodes, for the automatic design and update of sensor and information fusion systems. This article encompasses the fusion system configuration and adaptation as well as communication aspects. Manual interaction with the flexibly changing system is reduced to a minimum.

  6. An FPGA-based heterogeneous image fusion system design method

    Science.gov (United States)

    Song, Le; Lin, Yu-chi; Chen, Yan-hua; Zhao, Mei-rong

    2011-08-01

    Taking the advantages of FPGA's low cost and compact structure, an FPGA-based heterogeneous image fusion platform is established in this study. Altera's Cyclone IV series FPGA is adopted as the core processor of the platform, and the visible light CCD camera and infrared thermal imager are used as the image-capturing device in order to obtain dualchannel heterogeneous video images. Tailor-made image fusion algorithms such as gray-scale weighted averaging, maximum selection and minimum selection methods are analyzed and compared. VHDL language and the synchronous design method are utilized to perform a reliable RTL-level description. Altera's Quartus II 9.0 software is applied to simulate and implement the algorithm modules. The contrast experiments of various fusion algorithms show that, preferably image quality of the heterogeneous image fusion can be obtained on top of the proposed system. The applied range of the different fusion algorithms is also discussed.

  7. Measles Virus Fusion Protein: Structure, Function and Inhibition

    Directory of Open Access Journals (Sweden)

    Philippe Plattet

    2016-04-01

    Full Text Available Measles virus (MeV, a highly contagious member of the Paramyxoviridae family, causes measles in humans. The Paramyxoviridae family of negative single-stranded enveloped viruses includes several important human and animal pathogens, with MeV causing approximately 120,000 deaths annually. MeV and canine distemper virus (CDV-mediated diseases can be prevented by vaccination. However, sub-optimal vaccine delivery continues to foster MeV outbreaks. Post-exposure prophylaxis with antivirals has been proposed as a novel strategy to complement vaccination programs by filling herd immunity gaps. Recent research has shown that membrane fusion induced by the morbillivirus glycoproteins is the first critical step for viral entry and infection, and determines cell pathology and disease outcome. Our molecular understanding of morbillivirus-associated membrane fusion has greatly progressed towards the feasibility to control this process by treating the fusion glycoprotein with inhibitory molecules. Current approaches to develop anti-membrane fusion drugs and our knowledge on drug resistance mechanisms strongly suggest that combined therapies will be a prerequisite. Thus, discovery of additional anti-fusion and/or anti-attachment protein small-molecule compounds may eventually translate into realistic therapeutic options.

  8. Heterologous production of peptides in plants: fusion proteins and beyond.

    Science.gov (United States)

    Viana, Juliane Flávia Cançado; Dias, Simoni Campos; Franco, Octávio Luiz; Lacorte, Cristiano

    2013-11-01

    Recombinant DNA technology has allowed the ectopic production of proteins and peptides of different organisms leading to biopharmaceutical production in large cultures of bacterial, yeasts and mammalian cells. Otherwise, the expression of recombinant proteins and peptides in plants is an attractive alternative presenting several advantages over the commonly used expression systems including reduced production costs, easy scale-up and reduced risks of pathogen contamination. Different types of proteins and peptides have been expressed in plants, including antibodies, antigens, and proteins and peptides of medical, veterinary and industrial applications. However, apart from providing a proof of concept, the use of plants as platforms for heterologous protein and peptide production still depends on key steps towards optimization including the enhancement of expression levels, manipulation of post-transcriptional modifications and improvements in purification methods. In this review, strategies to increase heterologous protein and peptide stability and accumulation are discussed, focusing on the expression of peptides through the use of gene fusions.

  9. Localization of a region in the fusion protein of avian metapneumovirus that modulates cell-cell fusion.

    Science.gov (United States)

    Wei, Yongwei; Feng, Kurtis; Yao, Xiangjie; Cai, Hui; Li, Junan; Mirza, Anne M; Iorio, Ronald M; Li, Jianrong

    2012-11-01

    The genus Metapneumovirus within the subfamily Pneumovirinae of the family Paramyxoviridae includes two members, human metapneumovirus (hMPV) and avian metapneumovirus (aMPV), causing respiratory tract infections in humans and birds, respectively. Paramyxoviruses enter host cells by fusing the viral envelope with a host cell membrane. Membrane fusion of hMPV appears to be unique, in that fusion of some hMPV strains requires low pH. Here, we show that the fusion (F) proteins of aMPV promote fusion in the absence of the attachment protein and low pH is not required. Furthermore, there are notable differences in cell-cell fusion among aMPV subtypes. Trypsin was required for cell-cell fusion induced by subtype B but not subtypes A and C. The F protein of aMPV subtype A was highly fusogenic, whereas those from subtypes B and C were not. By construction and evaluation of chimeric F proteins composed of domains from the F proteins of subtypes A and B, we localized a region composed of amino acid residues 170 to 338 in the F protein that is responsible for the hyperfusogenic phenotype of the F from subtype A. Further mutagenesis analysis revealed that residues R295, G297, and K323 in this region collectively contributed to the hyperfusogenicity. Taken together, we have identified a region in the aMPV F protein that modulates the extent of membrane fusion. A model for fusion consistent with these data is presented.

  10. Present status of inertial confinement fusion reactor design

    International Nuclear Information System (INIS)

    Mima, Kunioki; Ido, Shunji; Nakai, Sadao.

    1986-01-01

    Since inertial nuclear fusion reactors do not require high vacuum and high magnetic field, the structure of the reactor cavity becomes markedly simple as compared with tokamak type fusion reactors. In particular, since high vacuum is not necessary, liquid metals such as lithium and lead can be used for the first wall, and the damage of reactor structures by neutrons can be prevented. As for the core, the energy efficiency of lasers is not very high, accordingly it must be designed so that the pellet gain due to nuclear fusion becomes sufficiently high, and typically, the gain coefficient from 100 to 200 is necessary. In this paper, the perspective of pellet gain, the plan from the present status to the practical reactors, and the conceptual design of the practical reactors are discussed. The plan of fuel ignition, energy break-even and high gain by the implosion mode, of which the uncertain factor due to uneven irradiation and instability was limited to the minimum, was clarified. The scenario of the development of laser nuclear fusion reactors is presented, and the concept of the reactor system is shown. The various types of nuclear fusion-fission hybrid reactors are explained. As for the design of inertial fusion power reactors, the engineering characteristics of the core, the conceptual design, water fall type reactors and DD fuel reactors are discussed. (Kako, I.)

  11. Fusion protein is the main determinant of metapneumovirus host tropism.

    Science.gov (United States)

    de Graaf, Miranda; Schrauwen, Eefje J A; Herfst, Sander; van Amerongen, Geert; Osterhaus, Albert D M E; Fouchier, Ron A M

    2009-06-01

    Human metapneumovirus (HMPV) and avian metapneumovirus subgroup C (AMPV-C) infect humans and birds, respectively. This study confirmed the difference in host range in turkey poults, and analysed the contribution of the individual metapneumovirus genes to host range in an in vitro cell-culture model. Mammalian Vero-118 cells supported replication of both HMPV and AMPV-C in contrast to avian quail fibroblast (QT6) cells in which only AMPV-C replicated to high titres. Inoculation of Vero-118 and QT6 cells with recombinant HMPV in which genes were exchanged with those of AMPV-C revealed that the metapneumovirus fusion (F) protein is the main determinant for host tropism. Chimeric viruses in which polymerase complex proteins were exchanged between HMPV and AMPV-C replicated less efficiently compared with HMPV in QT6 cells. Using mini-genome systems, it was shown that exchanging these polymerase proteins resulted in reduced replication and transcription efficiency in QT6 cells. Examination of infected Vero-118 and QT6 cells revealed that viruses containing the F protein of AMPV-C yielded larger syncytia compared with viruses containing the HMPV F protein. Cell-content mixing assays revealed that the F protein of AMPV-C was more fusogenic compared with the F protein of HMPV, and that the F2 region is responsible for the difference observed between AMPV-C and HMPV F-promoted fusion in QT6 and Vero-118 cells. This study provides insight into the determinants of host tropism and membrane fusion of metapneumoviruses.

  12. Identification of a human protein-derived HIV-1 fusion inhibitor targeting the gp41 fusion core structure.

    Directory of Open Access Journals (Sweden)

    Lijun Chao

    Full Text Available The HIV-1 envelope glycoprotein (Env gp41 plays a crucial role in the viral fusion process. The peptides derived from the C-terminal heptad repeat (CHR of gp41 are potent HIV fusion inhibitors. However, the activity of these anti-HIV-1 peptides in vivo may be attenuated by their induction of anti-gp41 antibodies. Thus, it is essential to identify antiviral peptides or proteins with low, or no, immunogenicity to humans. Here, we found that the C-terminal fragment (aa 462-521 of the human POB1 (the partner of RalBP1, designated C60, is an HIV-1 fusion inhibitor. It bound to N36, the peptide derived from the N-terminal heptad repeat (NHR of gp41, and to the six-helix bundle (6-HB formed by N36 and C34, a CHR-peptide, but it did not bind to C34. Unlike the CHR-peptides, C60 did not block gp41 6-HB formation. Rather, results suggest that C60 inhibits HIV-1 fusion by binding to the 6-HB, in particular, the residues in the gp41 NHR domain that are exposed on the surface of 6-HB. Since 6-HB plays a crucial role in the late stage of fusion between the viral envelope and endosomal membrane during the endocytic process of HIV-1, C60 may serve as a host restriction factor to suppress HIV-1 entry into CD4+ T lymphocytes. Taken together, it can be concluded from these results that C60 can be used as a lead for the development of anti-HIV-1 therapeutics or microbicides for the treatment and prevention of HIV-1 infection, as well as a molecular probe to study the fusogenic mechanism of HIV-1.

  13. Nuclear design of a very-low-activation fusion reactor

    International Nuclear Information System (INIS)

    Cheng, E.T.; Hopkins, G.R.

    1983-06-01

    An investigation was conducted to study the nuclear design aspects of using very-low-activation materials, such as SiC, MgO, and aluminum for fusion-reactor first wall, blanket, and shield applications. In addition to the advantage of very-low radioactive inventory, it was found that the very-low-activation fusion reactor can also offer an adequate tritium-breeding ratio and substantial amount of blanket nuclear heating as a conventional-material-structured reactor does. The most-stringent design constraint found in a very-low-activation fusion reactor is the limited space available in the inboard region of a tokamak concept for shielding to protect the superconducting toroidal field coil. A reference design was developed which mitigates the constraint by adopting a removable tungsten shield design that retains the inboard dimensions and gives the same shield performance as the reference STARFIRE tokamak reactor design

  14. Design study of an accelerator for heavy ion fusion

    International Nuclear Information System (INIS)

    Katayama, T.; Noda, A.; Tokuda, N.; Hirao, Y.

    1980-01-01

    Design of a demonstration accelerator for heavy ion fusion based on a synchrotron system is briefly described. The proposed complex system of injector linac, rapid cycling synchrotron and five accumulation rings can produce a peak current 1.6 kA, peak power 32 TW and total energy 0.3 MJ. Investigations of the intrabeam scattering give a lifetime of the beam longer than the fusion cycle time of 1 sec

  15. Definition and conceptual design of a small fusion reactor

    International Nuclear Information System (INIS)

    1979-04-01

    The objective of this project is to evaluate various mirror fusion reactor concepts that might result in small systems for the effective production of electrical power or stored energy (e.g., nuclear and chemical fuels). The basic two-year program goal is to select a particular concept and develop the conceptual design of a pilot plant that could provide a useful output from fusion. The pilot plant would be built and operated in the late 1980s

  16. Genetically engineered endostatin-lidamycin fusion proteins effectively inhibit tumor growth and metastasis

    International Nuclear Information System (INIS)

    Jiang, Wen-guo; Zhen, Yong-su; Lu, Xin-an; Shang, Bo-yang; Fu, Yan; Zhang, Sheng-hua; Zhou, Daifu; Li, Liang; Li, Yi; Luo, Yongzhang

    2013-01-01

    Endostatin (ES) inhibits endothelial cell proliferation, migration, invasion, and tube formation. It also shows antiangiogenesis and antitumor activities in several animal models. Endostatin specifically targets tumor vasculature to block tumor growth. Lidamycin (LDM), which consists of an active enediyne chromophore (AE) and a non-covalently bound apo-protein (LDP), is a member of chromoprotein family of antitumor antibiotics with extremely potent cytotoxicity to cancer cells. Therefore, we reasoned that endostatin-lidamycin (ES-LDM) fusion proteins upon energizing with enediyne chromophore may obtain the combined capability targeting tumor vasculature and tumor cell by respective ES and LDM moiety. In this study, we designed and obtained two new endostatin-based fusion proteins, endostatin-LDP (ES-LDP) and LDP-endostatin (LDP-ES). In vitro, the antiangiogenic effect of fusion proteins was determined by the wound healing assay and tube formation assay and the cytotoxicity of their enediyne-energized analogs was evaluated by CCK-8 assay. Tissue microarray was used to analyze the binding affinity of LDP, ES or ES-LDP with specimens of human lung tissue and lung tumor. The in vivo efficacy of the fusion proteins was evaluated with human lung carcinoma PG-BE1 xenograft and the experimental metastasis model of 4T1-luc breast cancer. ES-LDP and LDP-ES disrupted the formation of endothelial tube structures and inhibited endothelial cell migration. Evidently, ES-LDP accumulated in the tumor and suppressed tumor growth and metastasis. ES-LDP and ES show higher binding capability than LDP to lung carcinoma; in addition, ES-LDP and ES share similar binding capability. Furthermore, the enediyne-energized fusion protein ES-LDP-AE demonstrated significant efficacy against lung carcinoma xenograft in athymic mice. The ES-based fusion protein therapy provides some fundamental information for further drug development. Targeting both tumor vasculature and tumor cells by endostatin

  17. Design aspects of low activation fusion ignition experiments

    International Nuclear Information System (INIS)

    Cheng, E.T.; Creedon, R.L.; Hopkins, G.R.; Trester, P.W.; Wong, C.P.C.; Schultz, K.R.

    1986-01-01

    Preliminary design studies have been done exploring (1) materials selection, (2) shutdown biological dose rates, (3) mechanical design and (4) thermal design of a fusion ignition experiment made of low activation materials. From the results of these preliminary design studies it appears that an ignition experiment could be built of low activation materials, and that this design would allow hands-on access for maintenance

  18. Protein-induced fusion can be modulated by target membrane lipids through a structural switch at the level of the fusion peptide

    NARCIS (Netherlands)

    Pecheur, EI; Martin, [No Value; Bienvenue, A; Ruysschaert, JM; Hoekstra, D

    2000-01-01

    Regulatory features of protein-induced membrane fusion are largely unclear, particularly at the level of the fusion peptide. Fusion peptides being part of larger protein complexes, such investigations are met with technical limitations. Here, we show that the fusion activity of influenza virus or

  19. A fluorescent cassette-based strategy for engineering multiple domain fusion proteins

    Directory of Open Access Journals (Sweden)

    Khorchid Ahmad

    2003-07-01

    Full Text Available Abstract Background The engineering of fusion proteins has become increasingly important and most recently has formed the basis of many biosensors, protein purification systems, and classes of new drugs. Currently, most fusion proteins consist of three or fewer domains, however, more sophisticated designs could easily involve three or more domains. Using traditional subcloning strategies, this requires micromanagement of restriction enzymes sites that results in complex workaround solutions, if any at all. Results Therefore, to aid in the efficient construction of fusion proteins involving multiple domains, we have created a new expression vector that allows us to rapidly generate a library of cassettes. Cassettes have a standard vector structure based on four specific restriction endonuclease sites and using a subtle property of blunt or compatible cohesive end restriction enzymes, they can be fused in any order and number of times. Furthermore, the insertion of PCR products into our expression vector or the recombination of cassettes can be dramatically simplified by screening for the presence or absence of fluorescence. Conclusions Finally, the utility of this new strategy was demonstrated by the creation of basic cassettes for protein targeting to subcellular organelles and for protein purification using multiple affinity tags.

  20. Conceptual design study for a laser fusion hybrid

    International Nuclear Information System (INIS)

    Maniscalco, J.A.

    1976-01-01

    Lawrence Livermore Laboratory and Bechtel Corporation have been involved in a joint effort to conceptually design a laser fusion hybrid reactor. The design which has evolved is a depleted-uranium fueled fast-fission blanket which produces fissile plutonium and electricity. A major objective of the design study was to evaluate the feasibility of producing fissile fuel with laser fusion. This feasibility evaluation was carried out by analyzing the integrated engineering performance of the complete conceptual design and by identifying the required laser/pellet performance. The performance of the laser fusion hybrid has also been compared to a typical fast breeder reactor. The results show that the laser fusion hybrid produces enough fissile material to fuel more than six light water reactors (LWRs) of equivalent thermal power while operating in a regime which requires an order of magnitude less laser and pellet performance than pure laser fusion. In comparison to a fast breeder reactor the hybrid produces 10 times more fissile fuel. An economic analysis of the design shows that the cost of electricity in a combined hybrid-LWR scenario increases by only 20 to 40 percent when the capital cost of the hybrid ranges from 2 to 3 times more than an LWR

  1. Conceptual design study for a laser fusion hybrid

    International Nuclear Information System (INIS)

    Maniscalco, J.A.

    1976-09-01

    Lawrence Livermore Laboratory and Bechtel Corporation have been involved in a joint effort to conceptually design a laser fusion hybrid reactor. The design which has evolved is a depleted-uranium fueled fast-fission blanket which produces fissile plutonium and electricity. A major objective of the design study was to evaluate the feasibility of producing fissile fuel with laser fusion. This feasibility evaluation was carried out by analyzing the integrated engineering performance of the complete conceptual design and by identifying the required laser/pellet performance. The performance of the laser fusion hybrid has also been compared to a typical fast breeder reactor. The results show that the laser fusion hybrid produces enough fissile material to fuel more than six light water reactors (LWR's) of equivalent thermal power while operating in a regime which requires an order of magnitude less laser and pellet performance than pure laser fusion. In comparison to a fast breeder reactor the hybrid produces 10 times more fissile fuel. An economic analysis of the design shows that the cost of electricity in a combined hybrid-LWR scenario is insensitive to the capital cost of the hybrid, increasing by only 20 to 40 percent when the capital cost of the hybrid ranges from 2 to 3 times more than an LWR

  2. Research and Application of Autodesk Fusion360 in Industrial Design

    Science.gov (United States)

    Song, P. P.; Qi, Y. M.; Cai, D. C.

    2018-05-01

    In 2016, Fusion 360, a productintroduced byAutodesk and integrating industrial design, structural design, mechanical simulation, and CAM, turns out a design platform supportingcollaboration and sharing both cross-platform and via the cloud. In previous products, design and manufacturing use to be isolated. In the course of design, research and development, the communication between designers and engineers used to go on through different software products, tool commands, and even industry terms. Moreover, difficulty also lies with the communication between design thoughts and machining strategies. Naturally, a difficult product design and R & D process would trigger a noticeable gap between the design model and the actual product. A complete product development process tends to cover several major areas, such as industrial design, mechanical design, rendering and animation, computer aided emulation (CAE), and computer aided manufacturing (CAM). Fusion 360, a perfect design solving the technical problems of cross-platform data exchange, realizes the effective control of cross-regional collaboration and presents an overview of collaboration and breaks the barriers between art and manufacturing, andblocks between design and processing. The “Eco-development of Fusion360 Industrial Chain” is both a significant means to and an inevitable trend forthe manufacturers and industrial designers to carry out innovation in China.

  3. Safety issues relating to the design of fusion power facilities

    International Nuclear Information System (INIS)

    Stasko, R.R.; Wong, K.Y.; Russell, S.B.

    1986-06-01

    In order to make fusion power a viable future source of energy, it will be necessary to ensure that the cost of power for fusion electric generation is competitive with advanced fission concepts. In addition, fusion power will have to live up to its original promise of being a more radiologically benign technology than fission, and be able to demonstrate excellent operational safety performance. These two requirements are interrelated, since the selection of an appropriate safety philosophy early in the design phase could greatly reduce or eliminate the capital costs of elaborate safety related and protective sytems. This paper will briefly overview a few of the key safety issues presently recognized as critical to the ultimate achievement of licensable, environmentally safe and socially acceptable fusion power facilities. 12 refs

  4. ELMO Bumpy Torus fusion-reactor design study

    International Nuclear Information System (INIS)

    Bathke, C.G.; Krakowski, R.A.

    1981-01-01

    A complete power plant design of a 1200-MWe ELMO Bumpy Torus Reactor (EBTR) is described that emphasizes those features that are unique to the EBT confinement concept, with subsystems and balance-of-plant items that are generic to magnetic fusion being adopted from past, more extensive tokamak reactor designs

  5. Challenges of designing fusion reactors for remote maintainability

    International Nuclear Information System (INIS)

    Mason, L.S.

    1981-01-01

    One of the major problems faced by the fusion community is the development of the high level of reliability required to assure that fusion will be a viable commercial power source. Much of the responsibility for solving this problem falls directly on the designer in developing concepts that have a high level of maintainability. The problems are both near-term, in developing maintainability for next generation engineering oriented reactors; and long range, in developing full maintainability for the more commercial concepts with their required high level of on-line time. The near-time challenge will include development of unqiue design concepts to perform inspection, maintenance, replacement, and testing under the stringent conditions imposed by the next generation engineering oriented machines. The long range challenge will focus on basic design concepts that will enable the full mainatability required by commerical fusion

  6. Conceptual design study of a scyllac fusion test reactor

    International Nuclear Information System (INIS)

    Thomassen, K.I.

    1975-07-01

    The report describes a conceptual design study of a fusion test reactor based on the Scyllac toroidal theta-pinch approach to fusion. It is not the first attempt to describe the physics and technology required for demonstrating scientific feasibility of the approach, but it is the most complete design in the sense that the physics necessary to achieve the device goals is extrapolated from experimentally tested MHD theories of toroidal systems,and it uses technological systems whose engineering performance has been carefully calculated to ensure that they meet the machine requirements

  7. Challenges of designing fusion reactors for remote maintainability

    International Nuclear Information System (INIS)

    Masson, L.S.

    1981-01-01

    One of the major problems faced by the fusion community is the development of the high level of reliability required to assure that fusion will be a viable commercial power source. Much of the responsibility for solving this problem falls directly on the designer in developing concepts that have a high level of maintainability for the next generation engineering oriented reactors; and long range, in developing full maintainability for the more complicated commercial concepts with their required high level of on-line time. The near-term challenge will include development of unique design concepts to perform inspection, maintenance, replacement, and testing under the stringent conditions imposed by the next generation engineering oriented machines. The long range challenge will focus on basic design concepts that will enable the full maintainability required by commercial fusion. In addition to the purely technical challenges, the fusion community is also faced with the problem of developing programmatic means to assure that reactor maintenance issues are given proper and timely emphasis as the nuclear phase of fusion is approached

  8. Expression and Activation of Horseradish Peroxidase-Protein A/G Fusion Protein in Silkworm Larvae for Diagnostic Purposes.

    Science.gov (United States)

    Xxxx, Patmawati; Minamihata, Kosuke; Tatsuke, Tsuneyuki; Lee, Jae Man; Kusakabe, Takahiro; Kamiya, Noriho

    2018-06-01

    Recombinant protein production can create artificial proteins with desired functions by introducing genetic modifications to the target proteins. Horseradish peroxidase (HRP) has been used extensively as a reporter enzyme in biotechnological applications; however, recombinant production of HRP has not been very successful, hampering the utilization of HRP with genetic modifications. A fusion protein comprising an antibody binding protein and HRP will be an ideal bio-probe for high-quality HRP-based diagnostic systems. A HRP-protein A/G fusion protein (HRP-pAG) is designed and its production in silkworm (Bombyx mori) is evaluated for the first time. HRP-pAG is expressed in a soluble apo form, and is activated successfully by incubating with hemin. The activated HRP-pAG is used directly for ELISA experiments and retains its activity over 20 days at 4 °C. Moreover, HRP-pAG is modified with biotin by the microbial transglutaminase (MTG) reaction. The biotinylated HRP-pAG is conjugated with streptavidin to form a HRP-pAG multimer and the multimeric HRP-pAG produced higher signals in the ELISA system than monomeric HRP-pAG. The successful production of recombinant HRP in silkworm will contribute to creating novel HRP-based bioconjugates as well as further functionalization of HRP by applying enzymatic post-translational modifications. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  9. A Protein Disulfide Isomerase Gene Fusion Expression System That Increases the Extracellular Productivity of Bacillus brevis

    Science.gov (United States)

    Kajino, Tsutomu; Ohto, Chikara; Muramatsu, Masayoshi; Obata, Shusei; Udaka, Shigezo; Yamada, Yukio; Takahashi, Haruo

    2000-01-01

    We have developed a versatile Bacillus brevis expression and secretion system based on the use of fungal protein disulfide isomerase (PDI) as a gene fusion partner. Fusion with PDI increased the extracellular production of heterologous proteins (light chain of immunoglobulin G, 8-fold; geranylgeranyl pyrophosphate synthase, 12-fold). Linkage to PDI prevented the aggregation of the secreted proteins, resulting in high-level accumulation of fusion proteins in soluble and biologically active forms. We also show that the disulfide isomerase activity of PDI in a fusion protein is responsible for the suppression of the aggregation of the protein with intradisulfide, whereas aggregation of the protein without intradisulfide was prevented even when the protein was fused to a mutant PDI whose two active sites were disrupted, suggesting that another PDI function, such as chaperone-like activity, synergistically prevented the aggregation of heterologous proteins in the PDI fusion expression system. PMID:10653729

  10. Computational protein design: a review

    International Nuclear Information System (INIS)

    Coluzza, Ivan

    2017-01-01

    Proteins are one of the most versatile modular assembling systems in nature. Experimentally, more than 110 000 protein structures have been identified and more are deposited every day in the Protein Data Bank. Such an enormous structural variety is to a first approximation controlled by the sequence of amino acids along the peptide chain of each protein. Understanding how the structural and functional properties of the target can be encoded in this sequence is the main objective of protein design. Unfortunately, rational protein design remains one of the major challenges across the disciplines of biology, physics and chemistry. The implications of solving this problem are enormous and branch into materials science, drug design, evolution and even cryptography. For instance, in the field of drug design an effective computational method to design protein-based ligands for biological targets such as viruses, bacteria or tumour cells, could give a significant boost to the development of new therapies with reduced side effects. In materials science, self-assembly is a highly desired property and soon artificial proteins could represent a new class of designable self-assembling materials. The scope of this review is to describe the state of the art in computational protein design methods and give the reader an outline of what developments could be expected in the near future. (topical review)

  11. Conceptual design of the SlimCS fusion DEMO reactor

    International Nuclear Information System (INIS)

    Tobita, Kenji; Nishio, Satoshi; Enoeda, Mikio; Nakamura, Hirofumi; Hayashi, Takumi; Asakura, Nobuyuki; Utoh, Hiroyasu; Tanigawa, Hiroyasu; Nishitani, Takeo; Isono, Takaaki; Sakurai, Shinji; Kurita, Genichi; Hayashi, Takao; Oyama, Naoyuki; Liu Changle; Hamamatsu, Kiyotaka; Inoue, Takashi; Ozeki, Takahisa; Sato, Masayasu; Suzuki, Satoshi; Kawashima, Hisato; Ezato, Koichiro; Tsuru, Daigo; Koizumi, Norikiyo; Sakamoto, Keiji; Ando, Masami; Sakamoto, Yoshiteru; Shibama, Yusuke; Suzuki, Takahiro; Takechi, Manabu; Takahashi, Koji; Hirose, Takanori; Sato, Satoru; Nozawa, Takashi; Tanigawa, Hisashi; Kakudate, Satoshi; Kawamura, Yoshinori; Yamanishi, Toshihiko; Hoshino, Tsuyoshi; Ochiai, Kentaro; Ide, Shunsuke; Aiba, Nobuyuki; Shimizu, Katsuhiro; Honda, Mitsuru; Nakamichi, Masaru; Nishi, Hiroshi; Seki, Yoji; Nakamura, Yukiharu; Tsuchiya, Kunihiko; Yoshida, Tohru; Song Yuntao

    2010-08-01

    This report describes the results of the conceptual design study of the SlimCS fusion DEMO reactor aiming at demonstrating fusion power production in a plant scale and allowing to assess the economic prospects of a fusion power plant. The design study has focused on a compact and low aspect ratio tokamak reactor concept with a reduced-sized central solenoid, which is novel compared with previous tokamak reactor concept such as SSTR (Steady State Tokamak Reactor). Owing to low aspect ratio, the reactor will be capable of having comparatively high beta limit and high elongation (which can elevate the Greenwald density limit), having potential for high power density. The reactor has the main parameters of a major radius of 5.5 m, aspect ratio of 2.6, elongation of 2.0, normalized beta of 4.3, fusion out put of 2.95 GW and average neutron wall load of 3 MW/m 2 . This report covers various aspects of design study including systematic design, physics design, torus configuration, blanket, superconducting magnet, maintenance and building, which were carried out increase the engineering feasibility of the concept. (author)

  12. [Research progress in hirudin fusion protein--review].

    Science.gov (United States)

    Zhang, Chuan-Ling; Yu, Ai-Ping; Jin, Ji-De; Wu, Chu-Tse

    2007-02-01

    Natural hirudin extracted from the secretion of medical leech salivary gland is a single-chain peptide containing 65 aminoacid residues with molecular weight of 7000 D, and exists in three isomers of HV1, HV2 and HV3. Hirudin possesses three disulfide bridges forming the structure of core cyclic peptides, which binds to the catalytic site of thrombin so as to inhibit the catalysis of thrombin. Its c-terminus rich in acidic aminoacid residues possesses hydrophilicity, and is free on the molecular surface, and can bind with fibrin recognition site of hirudin. The minimal segment of 12 - 16 C-terminal acidic residues keeps the minimal activity of anti-thrombosis. Thus, hirudin, as a potent and specific inhibitor of thrombin, can be used to protect from and to treat clinically thrombosis. As it has some disadvantages such as short half-life, bleeding side-effect and mono-function, and so on, hirudin has been fused with some other functional proteins in recent years. The obtained fusion proteins can prolong the half life of hirudin, or relieve it bleeding side effect, or bring new functions, such as thrombolysis, inhibiting the platelet aggregation, targeting specifically. The research progress in hirudin fusion protein was summarized in this review.

  13. Conceptual design of a laser fusion power plant

    International Nuclear Information System (INIS)

    Maniscalco, J.A.; Meier, W.R.; Monsler, M.J.

    1977-01-01

    A conceptual design of a laser fusion power plant is extensively discussed. Recent advances in high gain targets are exploited in the design. A smaller blanket structure is made possible by use of a thick falling region of liquid lithium for a first wall. Major design features of the plant, reactor, and laser systems are described. A parametric analysis of performance and cost vs. design parameters is presented to show feasible design points. A more definitive follow-on conceptual design study is planned

  14. Localization of a Region in the Fusion Protein of Avian Metapneumovirus That Modulates Cell-Cell Fusion

    Science.gov (United States)

    Wei, Yongwei; Feng, Kurtis; Yao, Xiangjie; Cai, Hui; Li, Junan; Mirza, Anne M.; Iorio, Ronald M.

    2012-01-01

    The genus Metapneumovirus within the subfamily Pneumovirinae of the family Paramyxoviridae includes two members, human metapneumovirus (hMPV) and avian metapneumovirus (aMPV), causing respiratory tract infections in humans and birds, respectively. Paramyxoviruses enter host cells by fusing the viral envelope with a host cell membrane. Membrane fusion of hMPV appears to be unique, in that fusion of some hMPV strains requires low pH. Here, we show that the fusion (F) proteins of aMPV promote fusion in the absence of the attachment protein and low pH is not required. Furthermore, there are notable differences in cell-cell fusion among aMPV subtypes. Trypsin was required for cell-cell fusion induced by subtype B but not subtypes A and C. The F protein of aMPV subtype A was highly fusogenic, whereas those from subtypes B and C were not. By construction and evaluation of chimeric F proteins composed of domains from the F proteins of subtypes A and B, we localized a region composed of amino acid residues 170 to 338 in the F protein that is responsible for the hyperfusogenic phenotype of the F from subtype A. Further mutagenesis analysis revealed that residues R295, G297, and K323 in this region collectively contributed to the hyperfusogenicity. Taken together, we have identified a region in the aMPV F protein that modulates the extent of membrane fusion. A model for fusion consistent with these data is presented. PMID:22915815

  15. Introduction of fusion driven subcritical system plasma design

    International Nuclear Information System (INIS)

    Bin Wu

    2003-01-01

    Fusion driven subcritical nuclear system (FDS) is a multifunctional hybrid reactor, which could breed nuclear fuel, transmute long-lived wastes, producing tritium and so on. This paper presents an introduction of FDS plasma design. Several different advance equilibrium configurations have been proposed and a 1.5-D discharge simulation of FDS was also present

  16. Reactor design considerations for inertial confinement fusion

    International Nuclear Information System (INIS)

    Booth, L.A.

    1979-01-01

    The most challenging reactor design consideration is protection of the cavity wall from the various energy forms as released by the pellet and as affected by the reaction-chamber phenomena. These phenomena depend on both the design and the yield of the pellet, as well as on ambient conditions in the chamber at the time of the pellet microexplosion. The effects on pellet energy-release mechanisms of various reaction chamber atmosphere options are summarized

  17. The MARVEL domain protein, Singles Bar, is required for progression past the pre-fusion complex stage of myoblast fusion.

    Science.gov (United States)

    Estrada, Beatriz; Maeland, Anne D; Gisselbrecht, Stephen S; Bloor, James W; Brown, Nicholas H; Michelson, Alan M

    2007-07-15

    Multinucleated myotubes develop by the sequential fusion of individual myoblasts. Using a convergence of genomic and classical genetic approaches, we have discovered a novel gene, singles bar (sing), that is essential for myoblast fusion. sing encodes a small multipass transmembrane protein containing a MARVEL domain, which is found in vertebrate proteins involved in processes such as tight junction formation and vesicle trafficking where--as in myoblast fusion--membrane apposition occurs. sing is expressed in both founder cells and fusion competent myoblasts preceding and during myoblast fusion. Examination of embryos injected with double-stranded sing RNA or embryos homozygous for ethane methyl sulfonate-induced sing alleles revealed an identical phenotype: replacement of multinucleated myofibers by groups of single, myosin-expressing myoblasts at a stage when formation of the mature muscle pattern is complete in wild-type embryos. Unfused sing mutant myoblasts form clusters, suggesting that early recognition and adhesion of these cells are unimpaired. To further investigate this phenotype, we undertook electron microscopic ultrastructural studies of fusing myoblasts in both sing and wild-type embryos. These experiments revealed that more sing mutant myoblasts than wild-type contain pre-fusion complexes, which are characterized by electron-dense vesicles paired on either side of the fusing plasma membranes. In contrast, embryos mutant for another muscle fusion gene, blown fuse (blow), have a normal number of such complexes. Together, these results lead to the hypothesis that sing acts at a step distinct from that of blow, and that sing is required on both founder cell and fusion-competent myoblast membranes to allow progression past the pre-fusion complex stage of myoblast fusion, possibly by mediating fusion of the electron-dense vesicles to the plasma membrane.

  18. First preliminary design of an experimental fusion reactor

    International Nuclear Information System (INIS)

    1977-09-01

    A preliminary design of a tokamak experimental fusion reactor to be built in the near future is under way. The goals of the reactor are to achieve reactor-level plasma conditions for a sufficiently long operation period and to obtain design, construction and operational experience for the main components of full-scale power reactors. This design covers overall reactor system including plasma characteristics, reactor structure, blanket neutronics, shielding, superconducting magnets, neutral beam injector, electric power supply system, fuel circulating system, reactor cooling system, tritium recovery system and maintenance scheme. The main design parameters are as follows: the reactor fusion power 100 MW, torus radius 6.75 m, plasma radius 1.5 m, first wall radius 1.75 m, toroidal magnet field on axis 6 T, blanket fertile material Li 2 O, coolant He, structural material 316SS and tritium breeding ratio 0.9. (auth.)

  19. An Updated Point Design for Heavy Ion Fusion

    International Nuclear Information System (INIS)

    Yu, S.S.; Meier, W.R.; Abbott, R.B.; Barnard, J.J.; Brown, T.; Callahan, D.A.; Heitzenroeder, P.; Latkowski, J.F.; Logan, B.G.; Pemberton, S.J.; Peterson, P.F.; Rose, D.V.; Sabbi, G.L.; Sharp, W.M.; Welch, D.R.

    2002-01-01

    An updated, self-consistent point design for a heavy ion fusion (HIF) power plant based on an induction linac driver, indirect-drive targets, and a thick liquid wall chamber has been completed. Conservative parameters were selected to allow each design area to meet its functional requirements in a robust manner, and thus this design is referred to as the Robust Point Design (RPD-2002). This paper provides a top-level summary of the major characteristics and design parameters for the target, driver, final focus magnet layout and shielding, chamber, beam propagation to the target, and overall power plant

  20. Design study of blanket structure for tokamak experimental fusion reactor

    International Nuclear Information System (INIS)

    1979-11-01

    Design study of the blanket structure for JAERI Experimental Fusion Reactor (JXFR) has been carried out. Studied here were fabrication and testing of the blanket structure (blanket cells, blanket rings, piping and blanket modules), assembly and disassembly of the blanket module, and monitering and testing technique. Problems in design and fabrication of the blanket structure could be revealed. Research and development problems for the future were also disclosed. (author)

  1. Osiris and SOMBRERO inertial confinement fusion power plant designs

    International Nuclear Information System (INIS)

    Meier, W.R.; Bieri, R.L.; Monsler, M.J.

    1992-03-01

    Conceptual designs and assessments have been completed for two inertial fusion energy (IFE) electric power plants. The detailed designs and results of the assessment studies are presented in this report. Osiris is a heavy-ion-beam (HIB) driven power plant and SOMBRERO is a Krypton-Fluoride (KrF) laser-driven power plant. Both plants are sized for a net electric power of 1000 MWe

  2. Pooled-matrix protein interaction screens using Barcode Fusion Genetics.

    Science.gov (United States)

    Yachie, Nozomu; Petsalaki, Evangelia; Mellor, Joseph C; Weile, Jochen; Jacob, Yves; Verby, Marta; Ozturk, Sedide B; Li, Siyang; Cote, Atina G; Mosca, Roberto; Knapp, Jennifer J; Ko, Minjeong; Yu, Analyn; Gebbia, Marinella; Sahni, Nidhi; Yi, Song; Tyagi, Tanya; Sheykhkarimli, Dayag; Roth, Jonathan F; Wong, Cassandra; Musa, Louai; Snider, Jamie; Liu, Yi-Chun; Yu, Haiyuan; Braun, Pascal; Stagljar, Igor; Hao, Tong; Calderwood, Michael A; Pelletier, Laurence; Aloy, Patrick; Hill, David E; Vidal, Marc; Roth, Frederick P

    2016-04-22

    High-throughput binary protein interaction mapping is continuing to extend our understanding of cellular function and disease mechanisms. However, we remain one or two orders of magnitude away from a complete interaction map for humans and other major model organisms. Completion will require screening at substantially larger scales with many complementary assays, requiring further efficiency gains in proteome-scale interaction mapping. Here, we report Barcode Fusion Genetics-Yeast Two-Hybrid (BFG-Y2H), by which a full matrix of protein pairs can be screened in a single multiplexed strain pool. BFG-Y2H uses Cre recombination to fuse DNA barcodes from distinct plasmids, generating chimeric protein-pair barcodes that can be quantified via next-generation sequencing. We applied BFG-Y2H to four different matrices ranging in scale from ~25 K to 2.5 M protein pairs. The results show that BFG-Y2H increases the efficiency of protein matrix screening, with quality that is on par with state-of-the-art Y2H methods. © 2016 The Authors. Published under the terms of the CC BY 4.0 license.

  3. Design, construction, and characterization of high-performance membrane fusion devices with target-selectivity.

    Science.gov (United States)

    Kashiwada, Ayumi; Yamane, Iori; Tsuboi, Mana; Ando, Shun; Matsuda, Kiyomi

    2012-01-31

    Membrane fusion proteins such as the hemagglutinin glycoprotein have target recognition and fusion accelerative domains, where some synergistically working elements are essential for target-selective and highly effective native membrane fusion systems. In this work, novel membrane fusion devices bearing such domains were designed and constructed. We selected a phenylboronic acid derivative as a recognition domain for a sugar-like target and a transmembrane-peptide (Leu-Ala sequence) domain interacting with the target membrane, forming a stable hydrophobic α-helix and accelerating the fusion process. Artificial membrane fusion behavior between the synthetic devices in which pilot and target liposomes were incorporated was characterized by lipid-mixing and inner-leaflet lipid-mixing assays. Consequently, the devices bearing both the recognition and transmembrane domains brought about a remarkable increase in the initial rate for the membrane fusion compared with the devices containing the recognition domain alone. In addition, a weakly acidic pH-responsive device was also constructed by replacing three Leu residues in the transmembrane-peptide domain by Glu residues. The presence of Glu residues made the acidic pH-dependent hydrophobic α-helix formation possible as expected. The target-selective liposome-liposome fusion was accelerated in a weakly acidic pH range when the Glu-substituted device was incorporated in pilot liposomes. The use of this pH-responsive device seems to be a potential strategy for novel applications in a liposome-based delivery system. © 2011 American Chemical Society

  4. Conceptual design of the JAERI demonstration fusion reactor

    International Nuclear Information System (INIS)

    Sako, K.; Tone, T.; Seki, Y.

    1976-01-01

    Conceptual design of a tokamak demonstration fusion reactor is carried out. This design is an extended and improved version of the previous design which was presented at the 5th IAEA Conference. The main design parameters are as follows: the reactor thermal power 2000 MW, torus radius 10.5 m, plasma radius 2.7 m, first wall radius 3.0 m, toroidal magnetic field on axis 6T, blanket fertile material Li 2 O, coolant He, structural material Mo-alloy and tritium breeding ratio 1.2

  5. Mission and design of the Fusion Ignition Research Experiment (FIRE)

    International Nuclear Information System (INIS)

    Meade, D.M.; Jardin, S.C.; Schmidt, J.

    2001-01-01

    Experiments are needed to test and extend present understanding of confinement, macroscopic stability, alpha-driven instabilities, and particle/power exhaust in plasmas dominated by alpha heating. A key issue is to what extent pressure profile evolution driven by strong alpha heating will act to self-organize advanced configurations with large bootstrap current fractions and internal transport barriers. A design study of a Fusion Ignition Research Experiment (FIRE) is underway to assess near term opportunities for advancing the scientific understanding of self-heated fusion plasmas. The emphasis is on understanding the behavior of fusion plasmas dominated by alpha heating (Q≥5) that are sustained for durations comparable to the characteristic plasma time scales (≥20 τ E and ∼τ skin , where τ skin is the time for the plasma current profile to redistribute at fixed current). The programmatic mission of FIRE is to attain, explore, understand and optimize alpha-dominated plasmas to provide knowledge for the design of attractive magnetic fusion energy systems. The programmatic strategy is to access the alpha-heating-dominated regime with confidence using the present advanced tokamak data base (e.g., Elmy-H-mode, ≤0.75 Greenwald density) while maintaining the flexibility for accessing and exploring other advanced tokamak modes (e. g., reversed shear, pellet enhanced performance) at lower magnetic fields and fusion power for longer durations in later stages of the experimental program. A major goal is to develop a design concept that could meet these physics objectives with a construction cost in the range of $1B. (author)

  6. Qualitative Reliability Issues for Solid and Liquid Wall Fusion Design

    Energy Technology Data Exchange (ETDEWEB)

    Cadwallader, Lee Charles

    2001-01-01

    This report is an initial effort to identify issues affecting reliability and availability of solid and liquid wall designs for magnetic fusion power plant designs. A qualitative approach has been used to identify the possible failure modes of major system components and their effects on the systems. A general set of design attributes known to affect the service reliability has been examined for the overview solid and liquid wall designs, and some specific features of good first wall design have been discussed and applied to these designs as well. The two generalized designs compare well in regard to these design attributes. The strengths and weaknesses of each design approach are seen in the comparison of specific features.

  7. Qualitative Reliability Issues for Solid and Liquid Wall Fusion Designs

    Energy Technology Data Exchange (ETDEWEB)

    Cadwallader, L.C.

    2001-01-31

    This report is an initial effort to identify issues affecting reliability and availability of solid and liquid wall designs for magnetic fusion power plant designs. A qualitative approach has been used to identify the possible failure modes of major system components and their effects on the systems. A general set of design attributes known to affect the service reliability has been examined for the overview solid and liquid wall designs, and some specific features of good first wall design have been discussed and applied to these designs as well. The two generalized designs compare well in regard to these design attributes. The strengths and weaknesses of each design approach are seen in the comparison of specific features.

  8. Qualitative Reliability Issues for Solid and Liquid Wall Fusion Designs

    International Nuclear Information System (INIS)

    Cadwallader, L.C.

    2001-01-01

    This report is an initial effort to identify issues affecting reliability and availability of solid and liquid wall designs for magnetic fusion power plant designs. A qualitative approach has been used to identify the possible failure modes of major system components and their effects on the systems. A general set of design attributes known to affect the service reliability has been examined for the overview solid and liquid wall designs, and some specific features of good first wall design have been discussed and applied to these designs as well. The two generalized designs compare well in regard to these design attributes. The strengths and weaknesses of each design approach are seen in the comparison of specific features

  9. Fluorescent IgG fusion proteins made in E. coli

    Science.gov (United States)

    Luria, Yael; Raichlin, Dina; Benhar, Itai

    2012-01-01

    Antibodies are among the most powerful tools in biological and biomedical research and are presently the fastest growing category of new bio-pharmaceutics. The most common format of antibody applied for therapeutic, diagnostic and analytical purposes is the IgG format. For medical applications, recombinant IgGs are made in cultured mammalian cells in a process that is too expensive to be considered for producing antibodies for diagnostic and analytical purposes. Therefore, for such purposes, mouse monoclonal antibodies or polyclonal sera from immunized animals are used. While looking for an easier and more rapid way to prepare full-length IgGs for therapeutic purposes, we recently developed and reported an expression and purification protocol for full-length IgGs, and IgG-based fusion proteins in E. coli, called “Inclonals.” By applying the Inclonals technology, we could generate full-length IgGs that are genetically fused to toxins. The aim of the study described herein was to evaluate the possibility of applying the “Inclonals” technology for preparing IgG-fluorophore fusion proteins. We found that IgG fused to the green fluorescent proteins enhanced GFP (EGFP) while maintaining functionality in binding, lost most of its fluorescence during the refolding process. In contrast, we found that green fluorescent Superfolder GFP (SFGFP)-fused IgG and red fluorescent mCherry-fused IgG were functional in antigen binding and maintained fluorescence intensity. In addition, we found that we can link several SFGFPs in tandem to each IgG, with fluorescence intensity increasing accordingly. Fluorescent IgGs made in E. coli may become attractive alternatives to monoclonal or polyclonal fluorescent antibodies derived from animals. PMID:22531449

  10. Neutronic Parametric Study on a Conceptual Design for a Transmutation Fusion Blanket

    International Nuclear Information System (INIS)

    Tariq Siddique, M.; Kim, Myung Hyun

    2011-01-01

    Fusion energy may be the one of options of future energy. In all over the world, researchers are putting their efforts for its commercial and economical availability. Fusion-fission hybrid reactors have been studied for various applications in China. First milestone of fusion energy is expected to be the fusion fission hybrid reactors. In fusion-fission hybrid reactor the blanket design is of second prime importance after fusion source. In this study conceptual design of a fusion blanket is initiated for calculation of tritium production, transmutation of minor actinides (MA) and fission products (FP) and energy multiplication calculations

  11. Expression, purification and characterization of hepatitis B virus X protein BH3-like motif-linker-Bcl-xL fusion protein for structural studies

    Directory of Open Access Journals (Sweden)

    Hideki Kusunoki

    2017-03-01

    Full Text Available Hepatitis B virus X protein (HBx is a multifunctional protein that interacts directly with many host proteins. For example, HBx interacts with anti-apoptotic proteins, Bcl-2 and Bcl-xL, through its BH3-like motif, which leads to elevated cytosolic calcium levels, efficient viral DNA replication and the induction of apoptosis. To facilitate sample preparation and perform detailed structural characterization of the complex between HBx and Bcl-xL, we designed and purified a recombinant HBx BH3-like motif-linker-Bcl-xL fusion protein produced in E. coli. The fusion protein was characterized by size exclusion chromatography, circular dichroism and nuclear magnetic resonance experiments. Our results show that the fusion protein is a monomer in aqueous solution, forms a stable intramolecular complex, and likely retains the native conformation of the complex between Bcl-xL and the HBx BH3-like motif. Furthermore, the HBx BH3-like motif of the intramolecular complex forms an α-helix. These observations indicate that the fusion protein should facilitate structural studies aimed at understanding the interaction between HBx and Bcl-xL at the atomic level.

  12. Tritium management in fusion synfuel designs

    International Nuclear Information System (INIS)

    Galloway, T.R.

    1980-01-01

    Two blanket types are being studied: a lithium-sodium pool boiler and a lithium-oxide- or lithium-sodium pool boiler and a lithium-oxide- or aluminate-microsphere moving bed. For each, a wide variety of current technology was considered in handling the tritium. Here, we show the pool boiler with the sulfur-iodine thermochemical cycle first developed and now being piloted by the General Atomic Company. The tritium (T 2 ) will be generated in the lithium-sodium mixture where the concentration is approx. 10 ppM and held constant by a scavenging system consisting mainly of permeators. An intermediate sodium loop carries the blanket heat to the thermochemical cycle, and the T 2 in this loop is held to 1 ppM by a similar scavenging system. With this design, we have maintained blanket inventory at 1 kg of tritium, kept thermochemical cycle losses to 5 Ci/d and environmental loss to 10 Ci/d, and held total plant risk inventory at 7 kg tritium

  13. Fusion reactor design and technology program in China

    International Nuclear Information System (INIS)

    Huang, J.H.

    1994-01-01

    A fusion-fission hybrid reactor program was launched in 1987. The purpose of development of the hybrid reactor is twofold: to solve the problem of nuclear fuel supply for an expected large-scale development of fission reactor plants, and to maintain the momentum of fusion research. The program is described and the activities and progress of the program are presented. Two conceptual designs of an engineering test reactor with tokamak configuration were developed at the Southwestern Institute of Physics and the Institute of Plasma Physics. The results are a tokamak engineering test breeder (TETB) series design and a fusion-fission hybrid reactor design (SSEHR), characterized by a liquid-Li self-cooled blanket and an He-cooled solid tritium breeder blanket respectively. In parallel with the design studies, relevant technological experiments on a small or medium scale have been supported by this program. These include LHCD, ICRH and pellet injection in the area of plasma engineering; neutronics integral experiments with U, Pu, Fe and Be; various irradiation tests of austenitic and ferritic steels, magnetohydrodynamic (MHD) pressure drop experiments using a liquid metal loop; research into permeation barriers for tritium and hydrogen isotopes; solid tritium breeder tests using an in-situ loop in a fission reactor. All these experiments have proceeded successfully. The second step of this program is now starting. It seems reasonable that most of the research carried out in the first step will continue. ((orig.))

  14. Protein functional links in Trypanosoma brucei, identified by gene fusion analysis

    Directory of Open Access Journals (Sweden)

    Trimpalis Philip

    2011-07-01

    Full Text Available Abstract Background Domain or gene fusion analysis is a bioinformatics method for detecting gene fusions in one organism by comparing its genome to that of other organisms. The occurrence of gene fusions suggests that the two original genes that participated in the fusion are functionally linked, i.e. their gene products interact either as part of a multi-subunit protein complex, or in a metabolic pathway. Gene fusion analysis has been used to identify protein functional links in prokaryotes as well as in eukaryotic model organisms, such as yeast and Drosophila. Results In this study we have extended this approach to include a number of recently sequenced protists, four of which are pathogenic, to identify fusion linked proteins in Trypanosoma brucei, the causative agent of African sleeping sickness. We have also examined the evolution of the gene fusion events identified, to determine whether they can be attributed to fusion or fission, by looking at the conservation of the fused genes and of the individual component genes across the major eukaryotic and prokaryotic lineages. We find relatively limited occurrence of gene fusions/fissions within the protist lineages examined. Our results point to two trypanosome-specific gene fissions, which have recently been experimentally confirmed, one fusion involving proteins involved in the same metabolic pathway, as well as two novel putative functional links between fusion-linked protein pairs. Conclusions This is the first study of protein functional links in T. brucei identified by gene fusion analysis. We have used strict thresholds and only discuss results which are highly likely to be genuine and which either have already been or can be experimentally verified. We discuss the possible impact of the identification of these novel putative protein-protein interactions, to the development of new trypanosome therapeutic drugs.

  15. Engineering a pharmacologically superior form of granulocyte-colony-stimulating factor by fusion with gelatin-like-protein polymer.

    Science.gov (United States)

    Huang, Yan-Shan; Wen, Xiao-Fang; Wu, Yi-Liang; Wang, Ye-Fei; Fan, Min; Yang, Zhi-Yu; Liu, Wei; Zhou, Lin-Fu

    2010-03-01

    The plasma half-life of therapeutic proteins is a critical factor in many clinical applications. Therefore, new strategies to prolong plasma half-life of long-acting peptides and protein drugs are in high demand. Here, we designed an artificial gelatin-like protein (GLK) and fused this hydrophilic GLK polymer to granulocyte-colony-stimulating factor (G-CSF) to generate a chimeric GLK/G-CSF fusion protein. The genetically engineered recombinant GLK/G-CSF (rGLK/G-CSF) fusion protein was purified from Pichia pastoris. In vitro studies demonstrated that rGLK/G-CSF possessed an enlarged hydrodynamic radius, improved thermal stability and retained full bioactivity compared to unfused G-CSF. Following a single subcutaneous administration to rats, the rGLK/G-CSF fusion protein displayed a slower plasma clearance rate and stimulated greater and longer lasting increases in circulating white blood cells than G-CSF. Our findings indicate that fusion with this artificial, hydrophilic, GLK polymer provides many advantages in the construction of a potent hematopoietic factor with extended plasma half-life. This approach could be easily applied to other therapeutic proteins and have important clinical applications. (c) 2009 Elsevier B.V. All rights reserved.

  16. Tritium system design studies of fusion experimental breeder

    International Nuclear Information System (INIS)

    Deng Baiquan; Huang Jinhua

    2003-01-01

    A summary of the tritium system design studies for the engineering outline design of a fusion experimental breeder (FEB-E) is presented. This paper is divided into three sections. In first section, the geometry, loading features and tritium concentrations in liquid lithium of tritium breeding zones of blanket are described. The tritium flow chart corresponding to the tritium fuel cycle system has been constructed, and the inventories in ten subsystems are calculated using SWITRIM code in section 2. Results show that the necessary initial tritium storage to start up FEB-E with fusion power of 143 MW is about 319 g. In final section, the tritium leakage issues under different operation circumstances have been analyzed. It was found that the potential danger of tritium leakage could be resulted from the exhausted gas of the diverter system. It is important to elevate the tritium burnup fraction and reduce the tritium throughput. (authors)

  17. Conceptual design of laser fusion reactor KOYO-fast

    International Nuclear Information System (INIS)

    Tomabechi, K.; Kozaki, Y.; Norimatsu, T.

    2006-01-01

    A conceptual design of the laser fusion reactor KOYO-F based on the fast ignition scheme is reported including the target design, the laser system and the design for chamber. A Yb-YAG ceramic laser operated at 200 K is the primary candidate for the compression laser and an OPCPA (optical parametric chirped pulse amplification) system is the one for the ignition laser. The chamber is basically a wet wall type but the fire position is vertically off-set to simplify the protection scheme of the ceiling. The target consists of foam insulated, cryogenic DT shells with a LiPb, reentrant guide-cone. (authors)

  18. Second preliminary design of JAERI experimental fusion reactor (JXFR)

    International Nuclear Information System (INIS)

    Sako, Kiyoshi; Tone, Tatsuzo; Seki, Yasushi; Iida, Hiromasa; Yamato, Harumi

    1979-06-01

    Second preliminary design of a tokamak experimental fusion reactor to be built in the near future has been performed. This design covers overall reactor system including plasma characteristics, reactor structure, blanket neutronics radiation shielding, superconducting magnets, neutral beam injector, electric power supply system, fuel recirculating system, reactor cooling and tritium recovery systems and maintenance scheme. Safety analyses of the reactor system have been also performed. This paper gives a brief description of the design as of January, 1979. The feasibility study of raising the power density has been also studied and is shown as appendix. (author)

  19. Mechanical design of a magnetic fusion production reactor

    International Nuclear Information System (INIS)

    Neef, W.S.; Jassby, D.L.

    1986-01-01

    The mechanical aspects of a tandem mirror and tokamak concepts for the tritium production mission are compared, and a proposed breeding blanket configuration for each type of reactor is presented in detail, along with a design outline of the complete fusion reaction system. In both cases, the reactor design is developed sufficiently to permit preliminary cost estimates of all components. A qualitative comparison is drawn between both concepts from the view of mechanical design and serviceability, and suggestions are made for technology proof tests on unique mechanical features. Detailed cost breakdowns indicate less than 10% difference in the overall costs of the two reactors

  20. The coronavirus spike protein : mechanisms of membrane fusion and virion incorporation

    NARCIS (Netherlands)

    Bosch, B.J.

    2004-01-01

    The coronavirus spike protein is a membrane-anchored glycoprotein responsible for virus-cell attachment and membrane fusion, prerequisites for a successful virus infection. In this thesis, two aspects are described regarding the molecular biology of the coronavirus spike protein: its membrane fusion

  1. Spike Protein Fusion Peptide and Feline Coronavirus Virulence

    Science.gov (United States)

    Chang, Hui-Wen; Egberink, Herman F.; Halpin, Rebecca; Spiro, David J.

    2012-01-01

    Coronaviruses are well known for their potential to change their host or tissue tropism, resulting in unpredictable new diseases and changes in pathogenicity; severe acute respiratory syndrome and feline coronaviruses, respectively, are the most recognized examples. Feline coronaviruses occur as 2 pathotypes: nonvirulent feline enteric coronaviruses (FECVs), which replicate in intestinal epithelium cells, and lethal feline infectious peritonitis viruses (FIPVs), which replicate in macrophages. Evidence indicates that FIPV originates from FECV by mutation, but consistent distinguishing differences have not been established. We sequenced the full genome of 11 viruses of each pathotype and then focused on the single most distinctive site by additionally sequencing hundreds of viruses in that region. As a result, we identified 2 alternative amino acid differences in the putative fusion peptide of the spike protein that together distinguish FIPV from FECV in >95% of cases. By these and perhaps other mutations, the virus apparently acquires its macrophage tropism and spreads systemically. PMID:22709821

  2. Progress on the reference mirror fusion reactor design

    International Nuclear Information System (INIS)

    Carlson, G.A.; Doggett, J.N.; Moir, R.W.

    1976-01-01

    The design of a reference mirror fusion reactor is underway at Lawrence Livermore Laboratory. The reactor, rated at about 900 MWe, features steady-state operation, an absence of plasma impurity problems, and good accessibility for blanket maintenance. It is concluded that a mirror reactor appears workable, but its dollar/kWe cost will be considerably higher than present-day nuclear costs. The cost would be reduced most markedly by an increase in plasma Q

  3. Design issues for a laboratory high gain fusion facility

    International Nuclear Information System (INIS)

    Hogan, W.J.

    1987-01-01

    In an inertial fusion laboratory high gain facility, experiments will be carried out with up to 1000 MJ of thermonuclear yield. The experiment area of such a facility will include many systems and structures that will have to operate successfully in the difficult environment created by the sudden large energy release. This paper estimates many of the nuclear effects that will occur, discusses the implied design issues and suggests possible solutions so that a useful experimental facility can be built. 4 figs

  4. Menin-MLL inhibitors reverse oncogenic activity of MLL fusion proteins in leukemia.

    Science.gov (United States)

    Grembecka, Jolanta; He, Shihan; Shi, Aibin; Purohit, Trupta; Muntean, Andrew G; Sorenson, Roderick J; Showalter, Hollis D; Murai, Marcelo J; Belcher, Amalia M; Hartley, Thomas; Hess, Jay L; Cierpicki, Tomasz

    2012-01-29

    Translocations involving the mixed lineage leukemia (MLL) gene result in human acute leukemias with very poor prognosis. The leukemogenic activity of MLL fusion proteins is critically dependent on their direct interaction with menin, a product of the multiple endocrine neoplasia (MEN1) gene. Here we present what are to our knowledge the first small-molecule inhibitors of the menin-MLL fusion protein interaction that specifically bind menin with nanomolar affinities. These compounds effectively reverse MLL fusion protein-mediated leukemic transformation by downregulating the expression of target genes required for MLL fusion protein oncogenic activity. They also selectively block proliferation and induce both apoptosis and differentiation of leukemia cells harboring MLL translocations. Identification of these compounds provides a new tool for better understanding MLL-mediated leukemogenesis and represents a new approach for studying the role of menin as an oncogenic cofactor of MLL fusion proteins. Our findings also highlight a new therapeutic strategy for aggressive leukemias with MLL rearrangements.

  5. Osiris and SOMBRERO inertial confinement fusion power plant designs

    International Nuclear Information System (INIS)

    Meier, W.R.; Bieri, R.L.; Monsler, M.J.

    1992-03-01

    The primary objective of the of the IFE Reactor Design Studies was to provide the Office of Fusion Energy with an evaluation of the potential of inertial fusion for electric power production. The term reactor studies is somewhat of a misnomer since these studies included the conceptual design and analysis of all aspects of the IFE power plants: the chambers, heat transport and power conversion systems, other balance of plant facilities, target systems (including the target production, injection, and tracking systems), and the two drivers. The scope of the IFE Reactor Design Studies was quite ambitious. The majority of our effort was spent on the conceptual design of two IFE electric power plants, one using an induction linac heavy ion beam (HIB) driver and the other using a Krypton Fluoride (KrF) laser driver. After the two point designs were developed, they were assessed in terms of their (1) environmental and safety aspects; (2) reliability, availability, and maintainability; (3) technical issues and technology development requirements; and (4) economics. Finally, we compared the design features and the results of the assessments for the two designs

  6. Chaperone activity of human small heat shock protein-GST fusion proteins.

    Science.gov (United States)

    Arbach, Hannah; Butler, Caley; McMenimen, Kathryn A

    2017-07-01

    Small heat shock proteins (sHsps) are a ubiquitous part of the machinery that maintains cellular protein homeostasis by acting as molecular chaperones. sHsps bind to and prevent the aggregation of partially folded substrate proteins in an ATP-independent manner. sHsps are dynamic, forming an ensemble of structures from dimers to large oligomers through concentration-dependent equilibrium dissociation. Based on structural studies and mutagenesis experiments, it is proposed that the dimer is the smallest active chaperone unit, while larger oligomers may act as storage depots for sHsps or play additional roles in chaperone function. The complexity and dynamic nature of their structural organization has made elucidation of their chaperone function challenging. HspB1 and HspB5 are two canonical human sHsps that vary in sequence and are expressed in a wide variety of tissues. In order to determine the role of the dimer in chaperone activity, glutathione-S-transferase (GST) was genetically linked as a fusion protein to the N-terminus regions of both HspB1 and HspB5 (also known as Hsp27 and αB-crystallin, respectively) proteins in order to constrain oligomer formation of HspB1 and HspB5, by using GST, since it readily forms a dimeric structure. We monitored the chaperone activity of these fusion proteins, which suggest they primarily form dimers and monomers and function as active molecular chaperones. Furthermore, the two different fusion proteins exhibit different chaperone activity for two model substrate proteins, citrate synthase (CS) and malate dehydrogenase (MDH). GST-HspB1 prevents more aggregation of MDH compared to GST-HspB5 and wild type HspB1. However, when CS is the substrate, both GST-HspB1 and GST-HspB5 are equally effective chaperones. Furthermore, wild type proteins do not display equal activity toward the substrates, suggesting that each sHsp exhibits different substrate specificity. Thus, substrate specificity, as described here for full-length GST

  7. Elastin-like-polypeptide based fusion proteins for osteogenic factor delivery in bone healing.

    Science.gov (United States)

    McCarthy, Bryce; Yuan, Yuan; Koria, Piyush

    2016-07-08

    Modern treatments of bone injuries and diseases are becoming increasingly dependent on the usage of growth factors to stimulate bone growth. Bone morphogenetic protein-2 (BMP-2), a potent osteogenic inductive protein, exhibits promising results in treatment models, but recently has had its practical efficacy questioned due to the lack of local retention, ectopic bone formation, and potentially lethal inflammation. Where a new delivery technique of the BMP-2 is necessary, here we demonstrate the viability of an elastin-like peptide (ELP) fusion protein containing BMP-2 for delivery of the BMP-2. This fusion protein retains the performance characteristics of both the BMP-2 and ELP. The fusion protein was found to induce osteogenic differentiation of mesenchymal stem cells as evidenced by the production of alkaline phosphatase and extracellular calcium deposits in response to treatment by the fusion protein. Retention of the ELPs inverse phase transition property has allowed for expression of the fusion protein within a bacterial host (such as Escherichia coli) and easy and rapid purification using inverse transition cycling. The fusion protein formed self-aggregating nanoparticles at human-body temperature. The data collected suggests the viability of these fusion protein nanoparticles as a dosage-efficient and location-precise noncytotoxic delivery vehicle for BMP-2 in bone treatment. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1029-1037, 2016. © 2016 American Institute of Chemical Engineers.

  8. Cloning, Expression and Purification of the Recombinant HIV-1 Tat-Nef Fusion Protein in Prokaryotic Expression System

    Directory of Open Access Journals (Sweden)

    Somayeh Kadkhodayan

    2016-07-01

    Full Text Available Abstract Background: Nef is one of the HIV-1 critical proteins, because it is essential for viral replication and AIDS disease progression and induction of immune response against it can partially inhibit viral infection. Moreover, a domain of the HIV-1 Trans-Activator of Transcription (Tat, 48-60 aa could act as a cell penetrating peptide (CPP. In current study, cloning and expression of Tat-Nef fusion protein was performed in E. coli for the first time. The protein expression was confirmed by western blot analysis and was purified using reverse staining method. Materials and Methods: In this experimental study, primarily, cloning of Tat-Nef fusion gene was done in pGEX6p2 expression vector. Then, the expression of Tat-Nef recombinat protein in E.coli BL21 (DE3 strain was performed by using IPTG inducer. The protein expression was confirmed by SDS-PAGE and western blotting using anti-Nef monoclonal antibody. Then, the recombinant fusion protein was purified from gel using reverse staining method. Results: The results of PCR analysis and enzyme digestion showed a clear band of ~ 726 bp in agarose gel indicating the correct Tat-Nef fusion cloning in pGEX6p2 prokaryotic expression vector. In addition, a 54 kDa band of Tat-Nef on SDS-PAGE revealed Tat-Nef protein expression that western blot analysis using anti-Nef monoclonal antibody confirmed it. Conclusion: The purified Tat-Nef recombinant fusion protein will be used as an antigen for protein vaccine design against HIV infection.

  9. New concepts for controlled fusion reactor blanket design

    International Nuclear Information System (INIS)

    Conn, R.W.; Kulcinski, G.L.; Avci, H.; El-Maghrabi, M.

    1975-01-01

    Several new concepts for fusion reactor blanket design based on the idea of shifting, or tailoring, the neutron spectrum incident on the first structural wall are presented. The spectral shifter is a nonstructural element which can be made of graphite, silicon carbide, or three dimensionally woven carbon fibers (and containing other materials as appropriate) placed between the neutron source and the first structural wall. The softened neutron spectrum incident on the structural components leads to lower gas production and atom displacement rates than in more standard fusion blanket designs. In turn, this results in longer anticipated lifetimes for the structural materials and can significantly reduce radioactivity and afterheat levels. In addition, the neutron spectrum in the first structural wall can be made to approach the flux shape in fast breeder reactors. Such spectral softening means that existing radiation facilities may be more profitably used to provide relevant materials radiation damage data for the structural materials in these fusion blanket designs. This general class of blanket concepts are referred to as internal spectral shifter and energy converter, or ISSEC concepts. These specific design concepts fall into three main categories: ISSEC/EB concepts based on utilizing existing designs which breed tritium behind the first structural wall; ISSEC/IB concepts based on breeding tritium inside the first vacuum wall; and ISSEC/Bu concepts based on using boron, carbon, and perhaps, beryllium to obtain an energy multiplier and converter design that does not attempt to breed tritium or utilize lithium. The detailed analyses relate specifically to the nuclear performance of ISSEC systems and to a discussion of materials radiation damage problems in the structural material.(U.S.)

  10. Magnet Design Considerations for Fusion Nuclear Science Facility

    Energy Technology Data Exchange (ETDEWEB)

    Zhai, Y. [Princeton Plasma Physics Lab. (PPPL), Princeton, NJ (United States); Kessel, C. [Princeton Plasma Physics Lab. (PPPL), Princeton, NJ (United States); El-Guebaly, L. [Univ. of Wisconsin, Madison, WI (United States) Fusion Technology Institute; Titus, P. [Princeton Plasma Physics Lab. (PPPL), Princeton, NJ (United States)

    2016-06-01

    The Fusion Nuclear Science Facility (FNSF) is a nuclear confinement facility that provides a fusion environment with components of the reactor integrated together to bridge the technical gaps of burning plasma and nuclear science between the International Thermonuclear Experimental Reactor (ITER) and the demonstration power plant (DEMO). Compared with ITER, the FNSF is smaller in size but generates much higher magnetic field, i.e., 30 times higher neutron fluence with three orders of magnitude longer plasma operation at higher operating temperatures for structures surrounding the plasma. Input parameters to the magnet design from system code analysis include magnetic field of 7.5 T at the plasma center with a plasma major radius of 4.8 m and a minor radius of 1.2 m and a peak field of 15.5 T on the toroidal field (TF) coils for the FNSF. Both low-temperature superconductors (LTS) and high-temperature superconductors (HTS) are considered for the FNSF magnet design based on the state-of-the-art fusion magnet technology. The higher magnetic field can be achieved by using the high-performance ternary restacked-rod process Nb3Sn strands for TF magnets. The circular cable-in-conduit conductor (CICC) design similar to ITER magnets and a high-aspect-ratio rectangular CICC design are evaluated for FNSF magnets, but low-activation-jacket materials may need to be selected. The conductor design concept and TF coil winding pack composition and dimension based on the horizontal maintenance schemes are discussed. Neutron radiation limits for the LTS and HTS superconductors and electrical insulation materials are also reviewed based on the available materials previously tested. The material radiation limits for FNSF magnets are defined as part of the conceptual design studies for FNSF magnets.

  11. Evaluation of divertor conceptual designs for a fusion power plant

    International Nuclear Information System (INIS)

    Ferrari, M.; Giancarli, L.; Kleefeldt, K.; Nardi, C.; Roedig, M.; Reimann, J.; Salavy, J.F.

    2001-01-01

    In the frame of the preliminary study of plants suitable for the energy production from the fusion power, particular emphasis has been given on the divertor studies. Since a significant percentage of the power generated from the fusion process is absorbed in the divertor, the thermal efficiency of the power conversion cycle requires a high coolant outlet temperature of the divertor, leading to solutions that are different from those adopted for the present experimental fusion plants. Therefore, copper alloys having extremely high thermal conductivity, cannot be used as structural material for this kind of devices. The most suitable coolants to be used in the divertor are water, helium and liquid metals. A conceptual design study has been developed for each of these three fluids, with the aim to evaluate the maximum allowable thermal flux at the divertor target plate and the R and D requirements for each solution. While a water-cooled divertor can be designed with a limited R and D effort, the development of helium or liquid metal cooled divertors requires a more engaging R and D program

  12. Design of force-cooled conductors for large fusion magnets

    Energy Technology Data Exchange (ETDEWEB)

    Dresner, L.; Lue, J.W.

    1977-01-01

    Conductors cooled by supercritical helium in forced convection are under active consideration for large toroidal fusion magnets. One of the central problems in designing such force cooled conductors is to maintain an adequate stability margin while keeping the pumping power tolerably low. A method has been developed for minimizing the pumping power for fixed stability by optimally choosing the matrix-to-superconductor and the metal-to-helium ratios. Such optimized conductors reduce pumping power requirements for fusion size magnets to acceptable limits. Furthermore, the mass flow and hence pumping losses can be varied through a magnet according to the local magnetic field and magnitude of desired stability margin. Force cooled conductors give flexibility in operation, permitting, for example, higher fields to be obtained than originally intended by lowering the bath temperature or increasing the pumping power or both. This flexibility is only available if the pumping power is low to begin with. Scaling laws for the pumping requirement and stability margin as functions of operating current density, number of strands and such physical parameters as stabilizer resistivity and critical current density, have been proved. Numerical examples will be given for design of conductors intended for use in large toroidal fusion magnet systems.

  13. Conceptual design study for a mirror fusion breeder

    International Nuclear Information System (INIS)

    Huang Jinhua; Deng Boquan; Li Guiqing

    1986-01-01

    A mirror fusion breeder, CHD, has been designed for providing plenty of nuclear fuel for light water reactors to meet the needs for rapid development of nuclear power in the first half of next century. The breeder is able to support the nuclear fuel needs for more than 10 LWRs of equal scale in power with fuel enriched directly in CHD without reprocessing. Measures are taken to flatten the power density distribution in the blanket so that fission is suppressed in the region close to the plasma, and by this way fuel production is enhanced for this direct enriched fusion breeder. In order to reduce the MHD pressure drop, LiPb flows in the blanket axially. Though the tritium inventory in the reactor is very low, special material and design have to be developed to reduce the permeation of tritium through the coolant pipes. The cost of electricity from the system, consisting of 11 LWR plants and one fusion breeder is predicted to be 1.05 times of that from a traditional LWR plant. This figure is insensitive both to the cost of CHD and its support ratio

  14. Design of force-cooled conductors for large fusion magnets

    International Nuclear Information System (INIS)

    Dresner, L.; Lue, J.W.

    1977-01-01

    Conductors cooled by supercritical helium in forced convection are under active consideration for large toroidal fusion magnets. One of the central problems in designing such force cooled conductors is to maintain an adequate stability margin while keeping the pumping power tolerably low. A method has been developed for minimizing the pumping power for fixed stability by optimally choosing the matrix-to-superconductor and the metal-to-helium ratios. Such optimized conductors reduce pumping power requirements for fusion size magnets to acceptable limits. Furthermore, the mass flow and hence pumping losses can be varied through a magnet according to the local magnetic field and magnitude of desired stability margin. Force cooled conductors give flexibility in operation, permitting, for example, higher fields to be obtained than originally intended by lowering the bath temperature or increasing the pumping power or both. This flexibility is only available if the pumping power is low to begin with. Scaling laws for the pumping requirement and stability margin as functions of operating current density, number of strands and such physical parameters as stabilizer resistivity and critical current density, have been proved. Numerical examples will be given for design of conductors intended for use in large toroidal fusion magnet systems

  15. A Maltose-Binding Protein Fusion Construct Yields a Robust Crystallography Platform for MCL1.

    Directory of Open Access Journals (Sweden)

    Matthew C Clifton

    Full Text Available Crystallization of a maltose-binding protein MCL1 fusion has yielded a robust crystallography platform that generated the first apo MCL1 crystal structure, as well as five ligand-bound structures. The ability to obtain fragment-bound structures advances structure-based drug design efforts that, despite considerable effort, had previously been intractable by crystallography. In the ligand-independent crystal form we identify inhibitor binding modes not observed in earlier crystallographic systems. This MBP-MCL1 construct dramatically improves the structural understanding of well-validated MCL1 ligands, and will likely catalyze the structure-based optimization of high affinity MCL1 inhibitors.

  16. Critical plasma-materials issues for fusion reactor designs

    International Nuclear Information System (INIS)

    Wilson, K.L.; Bauer, W.

    1983-01-01

    Plasma-materials interactions are a dominant driving force in the design of fusion power reactors. This paper presents a summary of plasma-materials interactions research. Emphasis is placed on critical aspects related to reactor design. Particular issues to be addressed are plasma edge characterization, hydrogen recycle, impurity introduction, and coating development. Typical wall fluxes in operating magnetically confined devices are summarized. Recent calculations of tritium inventory and first wall permeation, based on laboratory measurements of hydrogen recycling, are given for various reactor operating scenarios. Impurity introduction/wall erosion mechanisms considered include sputtering, chemical erosion, and evaporation (melting). Finally, the advanced material development for in-vessel components is discussed. (author)

  17. Expression of the Acyl-Coenzyme A: Cholesterol Acyltransferase GFP Fusion Protein in Sf21 Insect Cells

    Science.gov (United States)

    Mahtani, H. K.; Richmond, R. C.; Chang, T. Y.; Chang, C. C. Y.; Rose, M. Franklin (Technical Monitor)

    2001-01-01

    The enzyme acyl-coenzyme A:cholesterol acyltransferase (ACAT) is an important contributor to the pathological expression of plaque leading to artherosclerosis n a major health problem. Adequate knowledge of the structure of this protein will enable pharmaceutical companies to design drugs specific to the enzyme. ACAT is a membrane protein located in the endoplasmic reticulum.t The protein has never been purified to homogeneity.T.Y. Chang's laboratory at Dartmouth College provided a 4-kb cDNA clone (K1) coding for a structural gene of the protein. We have modified the gene sequence and inserted the cDNA into the BioGreen His Baculovirus transfer vector. This was successfully expressed in Sf2l insect cells as a GFP-labeled ACAT protein. The advantage to this ACAT-GFP fusion protein (abbreviated GCAT) is that one can easily monitor its expression as a function of GFP excitation at 395 nm and emission at 509 nm. Moreover, the fusion protein GCAT can be detected on Western blots with the use of commercially available GFP antibodies. Antibodies against ACAT are not readily available. The presence of the 6xHis tag in the transfer vector facilitates purification of the recombinant protein since 6xHis fusion proteins bind with high affinity to Ni-NTA agarose. Obtaining highly pure protein in large quantities is essential for subsequent crystallization. The purified GCAT fusion protein can readily be cleaved into distinct GFP and ACAT proteins in the presence of thrombin. Thrombin digests the 6xHis tag linking the two protein sequences. Preliminary experiments have indicated that both GCAT and ACAT are expressed as functional proteins. The ultimate aim is to obtain large quantities of the ACAT protein in pure and functional form appropriate for protein crystal growth. Determining protein structure is the key to the design and development of effective drugs. X-ray analysis requires large homogeneous crystals that are difficult to obtain in the gravity environment of earth

  18. Design study of fusion Demo plant at JAERI

    International Nuclear Information System (INIS)

    Tobita, K.; Nishio, S.; Enoeda, M.

    2006-01-01

    Three options of fusion Demo plant are proposed characterized by functions of the center solenoid (Cs). The prime option uses a downsized CS, which does not provide sufficient V-s for plasma current ramp-up but supplies enough coil current for plasma shaping. This option produces a fusion output of 3 GW with a major radius of 5.5 m, aspect ratio of 2.6, normalized beta of 4.3 and maximum field of 16.4 T. The estimated reactor weight is lighter than that of other conventional tokamak reactors, suggesting an economic advantage. The plant uses rather conservative technologies such as Nb 3 Al superconductor, water-cooled solid breeder blanket, low activation ferritic steel as the structural material and tungsten monoblock divertor plate. The design philosophy and key issues related to the constituent technologies of the plant are described in the present paper

  19. Overview of the Fusion Engineering Device (FED) design

    International Nuclear Information System (INIS)

    Steiner, D.; Flanagan, C.A.

    1981-01-01

    The device has a major radius of 5.0 m with a plasma minor radius of 1.3 m elongated by 1.6. Capability is provided for operating the toroidal field coils up to 10 T, but the bulk of the operations are designed for 8 T. At 8-T conditions the fusion power is approx. 180 MW (neutron wall loading approx. 0.4 MW/m 2 ) and a plasma Q of approx. 5 is expected. At 10-T conditions, which are expected to be limited to about 10% of the total operations, the fusion power is approx. 450 MW (approx. 1.0 MW/m 2 ) and ignition is expected

  20. Overview of the fusion engineering device (FED) design

    International Nuclear Information System (INIS)

    Steiner, D.; Flanagan, C.A.

    1981-10-01

    The device has a major radius of 5.0 m with a plasma minor radius of 1.3 m elongated by 1.6. Capability is provided for operating the toroidal field coils up to 10 T, but the bulk of the operations are designed for 8 T. At 8-T conditions, the fusion power is approx. 180 MW (neutron wall loading approx. 0.4 MW/m 2 ) and a plasma Q of approx. 5 is expected. At 10-T conditions, which are expected to be limited to about 10% of the total operations, the fusion power is approx. 450 MW (approx. 1.0 MW/m 2 ) and ignition is expected

  1. Conceptual design of neutron diagnostic systems for fusion experimental reactor

    International Nuclear Information System (INIS)

    Iguchi, T.; Kaneko, J.; Nakazawa, M.

    1994-01-01

    Neutron measurement in fusion experimental reactors is very important for burning plasma diagnostics and control, monitoring of irradiation effects on device components, neutron source characterization for in-situ engineering tests, etc. A conceptual design of neutron diagnostic systems for an ITER-like fusion experimental reactor has been made, which consists of a neutron yield monitor, a neutron emission profile monitor and a 14-MeV spectrometer. Each of them is based on a unique idea to meet the required performances for full power conditions assumed at ITER operation. Micro-fission chambers of 235 U (and 238 U) placed at several poloidal angles near the first wall are adopted as a promising neutron yield monitor. A collimated long counter system using a 235 U fission chamber and graphite neutron moderators is also proposed to improve the calibration accuracy of absolute neutron yield determination

  2. Neutronics design for a spherical tokamak fusion-transmutation reactor

    International Nuclear Information System (INIS)

    Deng Meigen; Feng Kaiming; Yang Bangchao

    2002-01-01

    Based on studies of the spherical tokamak fusion reactors, a concept of fusion-transmutation reactor is put forward. By using the one-dimension transport and burn-up code BISON3.0 to process optimized design, a set of plasma parameters and blanket configuration suitable for the transmutation of MA (Minor Actinides) nuclear waste is selected. Based on the one-dimension calculation, two-dimension calculation has been carried out by using two-dimension neutronics code TWODANT. Combined with the neutron flux given by TWODANT calculation, burn-up calculation has been processed by using the one-dimension radioactivity calculation code FDKR and some useful and reasonable results are obtained

  3. Resolution of Disulfide Heterogeneity in Nogo Receptor 1 Fusion Proteins by Molecular Engineering

    Energy Technology Data Exchange (ETDEWEB)

    P Weinreb; D Wen; F Qian; C Wildes; E Garber; L Walus; M Jung; J Wang; J Relton; et al.

    2011-12-31

    NgRI (Nogo-66 receptor) is part of a signalling complex that inhibits axon regeneration in the central nervous system. Truncated soluble versions of NgRI have been used successfully to promote axon regeneration in animal models of spinal-cord injury, raising interest in this protein as a potential therapeutic target. The LRR (leucine-rich repeat) regions in NgRI are flanked by N- and C-terminal disulfide-containing 'cap' domains (LRRNT and LRRCT respectively). In the present work we show that, although functionally active, the NgRI(310)-Fc fusion protein contains mislinked and heterogeneous disulfide patterns in the LRRCT domain, and we report the generation of a series of variant molecules specifically designed to prevent this heterogeneity. Using these variants we explored the effects of modifying the NgRI truncation site or the spacing between the NgRI and Fc domains, or replacing cysteines within the NgRI or IgG hinge regions. One variant, which incorporates replacements of Cys{sup 266} and Cys{sup 309} with alanine residues, completely eliminated disulfide scrambling while maintaining functional in vitro and in vivo efficacy. This modified NgRI-Fc molecule represents a significantly improved candidate for further pharmaceutical development, and may serve as a useful model for the optimization of other IgG fusion proteins made from LRR proteins.

  4. Protein design for pathway engineering.

    Science.gov (United States)

    Eriksen, Dawn T; Lian, Jiazhang; Zhao, Huimin

    2014-02-01

    Design and construction of biochemical pathways has increased the complexity of biosynthetically-produced compounds when compared to single enzyme biocatalysis. However, the coordination of multiple enzymes can introduce a complicated set of obstacles to overcome in order to achieve a high titer and yield of the desired compound. Metabolic engineering has made great strides in developing tools to optimize the flux through a target pathway, but the inherent characteristics of a particular enzyme within the pathway can still limit the productivity. Thus, judicious protein design is critical for metabolic and pathway engineering. This review will describe various strategies and examples of applying protein design to pathway engineering to optimize the flux through the pathway. The proteins can be engineered for altered substrate specificity/selectivity, increased catalytic activity, reduced mass transfer limitations through specific protein localization, and reduced substrate/product inhibition. Protein engineering can also be expanded to design biosensors to enable high through-put screening and to customize cell signaling networks. These strategies have successfully engineered pathways for significantly increased productivity of the desired product or in the production of novel compounds. Copyright © 2013 Elsevier Inc. All rights reserved.

  5. Aequorin fusion proteins as bioluminescent tracers for competitive immunoassays

    Science.gov (United States)

    Mirasoli, Mara; Michelini, Elisa; Deo, Sapna K.; Dikici, Emre; Roda, Aldo; Daunert, Sylvia

    2004-06-01

    The use of bio- and chemiluminescence for the development of quantitative binding assays offers undoubted advantages over other detection systems, such as spectrophotometry, fluorescence, or radioactivity. Indeed, bio- and chemiluminescence detection provides similar, or even better, sensitivity and detectability than radioisotopes, while avoiding the problems of health hazards, waste disposal, and instability associated with the use of radioisotopes. Among bioluminescent labels, the calcium-activated photoprotein aequorin, originally isolated from Aequorea victoria and today available as a recombinant product, is characterized by very high detectability, down to attomole levels. It has been used as a bioluminescent label for developing a variety of highly sensitive immunoassays, using various analyte-aequorin conjugation strategies. When the analyte is a protein or a peptide, genetic engineering techniques can be used to produce protein fusions where the analyte is in-frame fused with aequorin, thus producing homogeneous one-to-one conjugation products, available in virtually unlimited amount. Various assays were developed using this strategy: a short review of the most interesting applications is presented, as well as the cloning, purification and initial characterization of an endothelin-1-aequorin conjugate suitable for developing a competitive immunoassay for endothelin-1, a potent vasoconstrictor peptide, involved in hypertension.

  6. OSIRIS and SOMBRERO Inertial Fusion Power Plant Designs, Volume 2: Designs, Assessments, and Comparisons

    Energy Technology Data Exchange (ETDEWEB)

    Meier, W. R.; Bieri, R. L.; Monsler, M. J.; Hendricks, C. D.; Laybourne, P.; Shillito, K. R.

    1992-03-01

    This is a comprehensive design study of two Inertial Fusion Energy (IFE) electric power plants. Conceptual designs are presented for a fusion reactor (called Osiris) using an induction-linac heavy-ion beam driver, and another (called SOMBRERO) using a KrF laser driver. The designs covered all aspects of IFE power plants, including the chambers, heat transport and power conversion systems, balance-of-plant facilities, target fabrication, target injection and tracking, as well as the heavy-ion and KrF drivers. The point designs were assessed and compared in terms of their environmental & safety aspects, reliability and availability, economics, and technology development needs.

  7. Study on conceptual design system of tritium production fusion reactor

    International Nuclear Information System (INIS)

    He Kaihui

    2004-11-01

    Conceptual design of an advanced tritium production reactor based on spherical torus, which is intermediate application of fusion energy, was presented. Different from traditional tokamak tritium production reactor design, advanced plasma physics performance and compact structural characteristics of ST were used to minimize tritium leakage and to maximize tritium breeding ratio with arrangement of tritium production blankets as possible as it can within vacuum vessel in order to produce 1 kg excess tritium except self-sufficient plasma core, corresponding plant availability 40% or more. Based on 2D neutronics calculation, preliminary conceptual design of ST-TPR was presented. Besides systematical analyses; design risk, uncertainty and backup are introduced generally for the backgrounds of next detailed conceptual design. (author)

  8. Conceptual design study of fusion experimental reactor (FER)

    International Nuclear Information System (INIS)

    1986-11-01

    Since 1980 the design study has been conducted at JAERI for the Fusion Experimental Reactor (FER) which has been proposed to be the next machine to JT-60 in the Japanese long term program of fusion reactor development. During two years from 1984 to 1985 FER concept was reviewed and redesigned. This report is the summary of the results obtained in the review and redesign activities in 1984 and 85. In the first year FER concept was discussed again and its frame work was reestablished. According to the new frame work the major reactor components of FER were designed. In the second year the whole plant system design including plant layout plan was conducted as well as the more detailed design analysis of the reactor conponents. The newly established frame for FER design is as follows: 1) Plasma : Self-ignition. 2) Operation scenario : Quasi-steady state operation with long burn pulse. 3) Neutron fluence on the first wall : 0.3 MWY/M 2 . 4) Blanket : Non-tritium breeding blanket with test modules for breeding blanket development. 5) Magnets : Superconducting Magnets. (author)

  9. Conceptual design study of fusion experimental reactor (FY86 FER)

    International Nuclear Information System (INIS)

    Kobayashi, Takeshi; Yamada, Masao; Mizoguchi, Tadanori

    1987-09-01

    This report describes the results of the reactor configuration/structure design for the fusion experimental reactor (FER) performed in FY 1986. The design was intended to meet the physical and engineering mission of the next step device which was decided by the subcommittee on the next step device of the nuclear fusion council. The objectives of the design study in FY 1986 are to advance and optimize the design concept of the last year because the recommendation of the subcommittee was basically the same as the design philosophy of the last year. Six candidate reactor configurations which correspond to options C ∼ D presented by the subcommittee were extensively examined. Consequently, ACS reactor (Advanced Option-C with Single Null Divertor) was selected as the reference configuration from viewpoints of technical risks and cost performance. Regarding the reactor structure, the following items were investigated intensively: minimization of reactor size, protection of first wall against plasma disruption, simplification of shield structure, reactor configuration which enables optimum arrangement of poloidal field coils. (author)

  10. Fusion

    CERN Document Server

    Mahaffey, James A

    2012-01-01

    As energy problems of the world grow, work toward fusion power continues at a greater pace than ever before. The topic of fusion is one that is often met with the most recognition and interest in the nuclear power arena. Written in clear and jargon-free prose, Fusion explores the big bang of creation to the blackout death of worn-out stars. A brief history of fusion research, beginning with the first tentative theories in the early 20th century, is also discussed, as well as the race for fusion power. This brand-new, full-color resource examines the various programs currently being funded or p

  11. Revealing Surface Waters on an Antifreeze Protein by Fusion Protein Crystallography Combined with Molecular Dynamic Simulations.

    Science.gov (United States)

    Sun, Tianjun; Gauthier, Sherry Y; Campbell, Robert L; Davies, Peter L

    2015-10-08

    Antifreeze proteins (AFPs) adsorb to ice through an extensive, flat, relatively hydrophobic surface. It has been suggested that this ice-binding site (IBS) organizes surface waters into an ice-like clathrate arrangement that matches and fuses to the quasi-liquid layer on the ice surface. On cooling, these waters join the ice lattice and freeze the AFP to its ligand. Evidence for the generality of this binding mechanism is limited because AFPs tend to crystallize with their IBS as a preferred protein-protein contact surface, which displaces some bound waters. Type III AFP is a 7 kDa globular protein with an IBS made up two adjacent surfaces. In the crystal structure of the most active isoform (QAE1), the part of the IBS that docks to the primary prism plane of ice is partially exposed to solvent and has clathrate waters present that match this plane of ice. The adjacent IBS, which matches the pyramidal plane of ice, is involved in protein-protein crystal contacts with few surface waters. Here we have changed the protein-protein contacts in the ice-binding region by crystallizing a fusion of QAE1 to maltose-binding protein. In this 1.9 Å structure, the IBS that fits the pyramidal plane of ice is exposed to solvent. By combining crystallography data with MD simulations, the surface waters on both sides of the IBS were revealed and match well with the target ice planes. The waters on the pyramidal plane IBS were loosely constrained, which might explain why other isoforms of type III AFP that lack the prism plane IBS are less active than QAE1. The AFP fusion crystallization method can potentially be used to force the exposure to solvent of the IBS on other AFPs to reveal the locations of key surface waters.

  12. Mechanical design aspects of a tandem mirror fusion reactor

    International Nuclear Information System (INIS)

    Neef, W.S. Jr.

    1977-01-01

    Two ''plugs'' of dense plasma at either end of a central solenoid cell form the basis of a new mirror fusion power plant concept. A central cell blanket design is presented. Modules on crawler tracks serviced by remote welding and handling machines of very simple design are important features resulting from linear axisymmetric geometry. Three blanket designs are considered and the best one presented in some detail. It has lithium as the breeder material, helium cooled. ''Plug'' magnet field strengths must be high. A novel magnet is presented to satisfy the physics of the end plugs. Beam sources at 1,200 KV present special problems. Methods of voltage standoff, arc damage control, and neutralization are discussed. New secondary containment ideas are presented to allow removable roof sections of balanced design

  13. Conceptual design study of fusion experimental reactor (FY86 FER)

    International Nuclear Information System (INIS)

    Saito, Ryusei; Kashihara, Shin-ichiro; Itoh, Shin-ichi

    1987-08-01

    This report describes the results of conceptual design study on plant systems for the Fusion Experimental Reactor (FY86 FER). Design studies for FER plant systems have been continued from FY85, especially for design modifications made in accordance with revisions of plasma scaling parameters and system improvements. This report describes 1) system construction, 2) site and reactor building plan, 3) repaire and maintenance system, 4) tritium circulation system, 5) heating, ventilation and air conditioning system, 6) tritium clean-up system, 7) cooling and baking system, 8) waste treatment and storage system, 9) control system, 10) electric power system, 11) site factory plan, all of which are a part of FY86 design work. The plant systems described in this report generally have been based on the FY86 FER (ACS Reactor) which is an one of the six candidates for FER. (author)

  14. Tritium pellet injector design for tokamak fusion test reactor

    International Nuclear Information System (INIS)

    Fisher, P.W.; Baylor, L.R.; Bryan, W.E.

    1985-01-01

    A tritium pellet injector (TPI) system has been designed for the Tokamak Fusion Test Reactor (TFTR) Q approx. 1 phase of operation. The injector gun utilizes a radial design with eight independent barrels and a common extruder to minimize tritium inventory. The injection line contains guide tubes with intermediate vacuum pumping stations and fast valves to minimize propellant leakage to the torus. The vacuum system is designed for tritium compatibility. The entire injector system is contained in a glove box for secondary containment protection against tritium release. Failure modes and effects have been analyzed, and structural analysis has been performed for most intense predicted earthquake conditions. Details of the design and operation of this system are presented in this paper

  15. Conceptual design report for a Fusion Engineering Device sector-handling machine and movable manipulator system

    International Nuclear Information System (INIS)

    Watts, K.D.; Masson, L.S.; McPherson, R.S.

    1982-10-01

    Design requirements, trade studies, design descriptions, conceptual designs, and cost estimates have been completed for the Fusion Engineering Device sector handling machine, movable manipulator system, subcomponent handling machine, and limiter blade handling machine. This information will be used by the Fusion Engineering Design Center to begin to determine the cost and magnitude of the effort required to perform remote maintenance on the Fusion Engineering Device. The designs presented are by no means optimum, and the costs estimates are rough-order-of-magnitude

  16. Protein design using continuous rotamers.

    Directory of Open Access Journals (Sweden)

    Pablo Gainza

    2012-01-01

    Full Text Available Optimizing amino acid conformation and identity is a central problem in computational protein design. Protein design algorithms must allow realistic protein flexibility to occur during this optimization, or they may fail to find the best sequence with the lowest energy. Most design algorithms implement side-chain flexibility by allowing the side chains to move between a small set of discrete, low-energy states, which we call rigid rotamers. In this work we show that allowing continuous side-chain flexibility (which we call continuous rotamers greatly improves protein flexibility modeling. We present a large-scale study that compares the sequences and best energy conformations in 69 protein-core redesigns using a rigid-rotamer model versus a continuous-rotamer model. We show that in nearly all of our redesigns the sequence found by the continuous-rotamer model is different and has a lower energy than the one found by the rigid-rotamer model. Moreover, the sequences found by the continuous-rotamer model are more similar to the native sequences. We then show that the seemingly easy solution of sampling more rigid rotamers within the continuous region is not a practical alternative to a continuous-rotamer model: at computationally feasible resolutions, using more rigid rotamers was never better than a continuous-rotamer model and almost always resulted in higher energies. Finally, we present a new protein design algorithm based on the dead-end elimination (DEE algorithm, which we call iMinDEE, that makes the use of continuous rotamers feasible in larger systems. iMinDEE guarantees finding the optimal answer while pruning the search space with close to the same efficiency of DEE.Software is available under the Lesser GNU Public License v3. Contact the authors for source code.

  17. Integrated Chamber Design for the Laser Inertial Fusion Energy (LIFE) Engine

    Energy Technology Data Exchange (ETDEWEB)

    Latkowski, J F; Kramer, K J; Abbott, R P; Morris, K R; DeMuth, J; Divol, L; El-Dasher, B; Lafuente, A; Loosmore, G; Reyes, S; Moses, G A; Fratoni, M; Flowers, D; Aceves, S; Rhodes, M; Kane, J; Scott, H; Kramer, R; Pantano, C; Scullard, C; Sawicki, R; Wilks, S; Mehl, M

    2010-12-07

    The Laser Inertial Fusion Energy (LIFE) concept is being designed to operate as either a pure fusion or hybrid fusion-fission system. A key component of a LIFE engine is the fusion chamber subsystem. The present work details the chamber design for the pure fusion option. The fusion chamber consists of the first wall and blanket. This integrated system must absorb the fusion energy, produce fusion fuel to replace that burned in previous targets, and enable both target and laser beam transport to the ignition point. The chamber system also must mitigate target emissions, including ions, x-rays and neutrons and reset itself to enable operation at 10-15 Hz. Finally, the chamber must offer a high level of availability, which implies both a reasonable lifetime and the ability to rapidly replace damaged components. An integrated LIFE design that meets all of these requirements is described herein.

  18. Integrated Chamber Design for the Laser Inertial Fusion Energy (LIFE) Engine

    International Nuclear Information System (INIS)

    Latkowski, J.F.; Kramer, K.J.; Abbott, R.P.; Morris, K.R.; DeMuth, J.; Divol, L.; El-Dasher, B.; Lafuente, A.; Loosmore, G.; Reyes, S.; Moses, G.A.; Fratoni, M.; Flowers, D.; Aceves, S.; Rhodes, M.; Kane, J.; Scott, H.; Kramer, R.; Pantano, C.; Scullard, C.; Sawicki, R.; Wilks, S.; Mehl, M.

    2010-01-01

    The Laser Inertial Fusion Energy (LIFE) concept is being designed to operate as either a pure fusion or hybrid fusion-fission system. A key component of a LIFE engine is the fusion chamber subsystem. The present work details the chamber design for the pure fusion option. The fusion chamber consists of the first wall and blanket. This integrated system must absorb the fusion energy, produce fusion fuel to replace that burned in previous targets, and enable both target and laser beam transport to the ignition point. The chamber system also must mitigate target emissions, including ions, x-rays and neutrons and reset itself to enable operation at 10-15 Hz. Finally, the chamber must offer a high level of availability, which implies both a reasonable lifetime and the ability to rapidly replace damaged components. An integrated LIFE design that meets all of these requirements is described herein.

  19. Application of uncertainty analysis in conceptual fusion reactor design

    International Nuclear Information System (INIS)

    Wu, T.; Maynard, C.W.

    1979-01-01

    The theories of sensitivity and uncertainty analysis are described and applied to a new conceptual tokamak fusion reactor design--NUWMAK. The responses investigated in this study include the tritium breeding ratio, first wall Ti dpa and gas productions, nuclear heating in the blanket, energy leakage to the magnet, and the dpa rate in the superconducting magnet aluminum stabilizer. The sensitivities and uncertainties of these responses are calculated. The cost/benefit feature of proposed integral measurements is also studied through the uncertainty reductions of these responses

  20. Shield design for the Fusion Materials Irradiation Test facility

    International Nuclear Information System (INIS)

    Carter, L.L.; Mann, F.M.; Morford, R.J.; Wilcox, A.D.; Johnson, D.L.; Huang, S.T.

    1983-03-01

    The shield design for the Fusion Materials Irradiation Test facility is based upon one-, two- and three-dimensional transport calculations with experimental measurements utilized to refine the nuclear data including the neutron cross sections from 20 to 50 MeV and the gamma ray and neutron source terms. The high energy neutrons and deuterons produce activation products from the numerous reactions that are kinematically allowed. The analyses for both beam-on and beam-off (from the activation products) conditions have required extensive nuclear data libraries and the utilization of Monte Carlo, discrete ordinates, point kernel and auxiliary computer codes

  1. Laser Intertial Fusion Energy: Neutronic Design Aspects of a Hybrid Fusion-Fission Nuclear Energy System

    Energy Technology Data Exchange (ETDEWEB)

    Kramer, Kevin James [Univ. of California, Berkeley, CA (United States)

    2010-04-08

    This study investigates the neutronics design aspects of a hybrid fusion-fission energy system called the Laser Fusion-Fission Hybrid (LFFH). A LFFH combines current Laser Inertial Confinement fusion technology with that of advanced fission reactor technology to produce a system that eliminates many of the negative aspects of pure fusion or pure fission systems. When examining the LFFH energy mission, a significant portion of the United States and world energy production could be supplied by LFFH plants. The LFFH engine described utilizes a central fusion chamber surrounded by multiple layers of multiplying and moderating media. These layers, or blankets, include coolant plenums, a beryllium (Be) multiplier layer, a fertile fission blanket and a graphite-pebble reflector. Each layer is separated by perforated oxide dispersion strengthened (ODS) ferritic steel walls. The central fusion chamber is surrounded by an ODS ferritic steel first wall. The first wall is coated with 250-500 μm of tungsten to mitigate x-ray damage. The first wall is cooled by Li17Pb83 eutectic, chosen for its neutron multiplication and good heat transfer properties. The Li17Pb83 flows in a jacket around the first wall to an extraction plenum. The main coolant injection plenum is immediately behind the Li17Pb83, separated from the Li17Pb83 by a solid ODS wall. This main system coolant is the molten salt flibe (2LiF-BeF2), chosen for beneficial neutronics and heat transfer properties. The use of flibe enables both fusion fuel production (tritium) and neutron moderation and multiplication for the fission blanket. A Be pebble (1 cm diameter) multiplier layer surrounds the coolant injection plenum and the coolant flows radially through perforated walls across the bed. Outside the Be layer, a fission fuel layer comprised of depleted uranium contained in Tristructural-isotropic (TRISO) fuel particles

  2. LIBRA - a light ion beam fusion conceptual reactor design

    International Nuclear Information System (INIS)

    Badger, B.; Moses, G.A.; Engelstad, R.L.; Kulcinski, G.L.; Lovell, E.; MacFarlane, J.; Peterson, R.R.; Sawan, M.E.; Sviatovslavsky, I.N.; Wittenberg, L.J.; Cook, D.L.; Olson, R.E.; Stinnett, R.W.; Ehrhardt, J.; Kessler, G.; Stein, E.

    1990-08-01

    The LIBRA light ion beam fusion commercial reactor study is a self-consistent conceptual design of a 330 MWe power plant with an accompanying economic analysis. Fusion targets are imploded by 4 MJ shaped pulses of 30 MeV Li ions at a rate of 3 Hz. The target gain is 80, leading to a yield of 320 MJ. The high intensity part of the ion pulse is delivered by 16 diodes through 16 separate z-pinch plasma channels formed in 100 torr of helium with trace amounts of lithium. The blanket is an array of porous flexible silicon carbind tubes with Li 17 Pb 83 flowing downward through them. These tubes (INPORT units) shield the target chamber wall from both neutron damage and the shock overpressure of the target explosion. The target chamber is 'self-pumped' by the target explosion generated overpressure into a surge tank partially filled with Li 17 Pb 83 that surrounds the target chamber. This scheme refreshes the chamber at the desired 3 Hz frequently without excessive pumping demands. The blanket multiplication is 1.2 and the tritium breeding ratio is 1.4. The direct capital cost of a 331 MWe LIBRA design is estimated to be 2843 Dollar/kWe while a 1200 MWe LIBRA design will cost approximately 1300 Dollar/kWe. (orig.) [de

  3. The Fusion Loops of the Initial Prefusion Conformation of Herpes Simplex Virus 1 Fusion Protein Point Toward the Membrane

    Directory of Open Access Journals (Sweden)

    Juan Fontana

    2017-08-01

    Full Text Available All enveloped viruses, including herpesviruses, must fuse their envelope with the host membrane to deliver their genomes into target cells, making this essential step subject to interference by antibodies and drugs. Viral fusion is mediated by a viral surface protein that transits from an initial prefusion conformation to a final postfusion conformation. Strikingly, the prefusion conformation of the herpesvirus fusion protein, gB, is poorly understood. Herpes simplex virus (HSV, a model system for herpesviruses, causes diseases ranging from mild skin lesions to serious encephalitis and neonatal infections. Using cryo-electron tomography and subtomogram averaging, we have characterized the structure of the prefusion conformation and fusion intermediates of HSV-1 gB. To this end, we have set up a system that generates microvesicles displaying full-length gB on their envelope. We confirmed proper folding of gB by nondenaturing electrophoresis-Western blotting with a panel of monoclonal antibodies (MAbs covering all gB domains. To elucidate the arrangement of gB domains, we labeled them by using (i mutagenesis to insert fluorescent proteins at specific positions, (ii coexpression of gB with Fabs for a neutralizing MAb with known binding sites, and (iii incubation of gB with an antibody directed against the fusion loops. Our results show that gB starts in a compact prefusion conformation with the fusion loops pointing toward the viral membrane and suggest, for the first time, a model for gB’s conformational rearrangements during fusion. These experiments further illustrate how neutralizing antibodies can interfere with the essential gB structural transitions that mediate viral entry and therefore infectivity.

  4. Nanoparticle-Fusion Protein Complexes Protect against Mycobacterium tuberculosis Infection.

    Science.gov (United States)

    Hart, Peter; Copland, Alastair; Diogo, Gil Reynolds; Harris, Shane; Spallek, Ralf; Oehlmann, Wulf; Singh, Mahavir; Basile, Juan; Rottenberg, Martin; Paul, Matthew John; Reljic, Rajko

    2018-03-07

    Tuberculosis (TB) is the leading cause of death from infectious disease, and the current vaccine, Bacillus Calmette-Guerin (BCG), is inadequate. Nanoparticles (NPs) are an emerging vaccine technology, with recent successes in oncology and infectious diseases. NPs have been exploited as antigen delivery systems and also for their adjuvantic properties. However, the mechanisms underlying their immunological activity remain obscure. Here, we developed a novel mucosal TB vaccine (Nano-FP1) based upon yellow carnauba wax NPs (YC-NPs), coated with a fusion protein consisting of three Mycobacterium tuberculosis (Mtb) antigens: Acr, Ag85B, and HBHA. Mucosal immunization of BCG-primed mice with Nano-FP1 significantly enhanced protection in animals challenged with low-dose, aerosolized Mtb. Bacterial control by Nano-FP1 was associated with dramatically enhanced cellular immunity compared to BCG, including superior CD4 + and CD8 + T cell proliferation, tissue-resident memory T cell (Trm) seeding in the lungs, and cytokine polyfunctionality. Alongside these effects, we also observed potent humoral responses, such as the generation of Ag85B-specific serum IgG and respiratory IgA. Finally, we found that YC-NPs were able to activate antigen-presenting cells via an unconventional IRF-3-associated activation signature, without the production of potentially harmful inflammatory mediators, providing a mechanistic framework for vaccine efficacy and future development. Copyright © 2017. Published by Elsevier Inc.

  5. Neutronics design aspects of reference ARIES-I fusion blanket

    International Nuclear Information System (INIS)

    Cheng, E.T.

    1990-12-01

    A SiC composite blanket concept was recently conceived for a deuterium-tritium burning, 1000 MW(e) tokamak fusion reactor design, ARIES-I. SiC composite structural material was chosen due to its very low activation features. High blanket nuclear performance and thermal efficiency, adequate tritium breeding, and a low level of activation are important design requirements for the ARIES-I reactor. The major approaches, other than using SiC as structural material, in meeting these design requirements, are to employ beryllium, the only low activation neutron multiplying material, and isotopically tailored Li 2 ZrO 3 , a tritium breeding material stable at high temperature, as blanket materials. 5 refs., 4 figs., 2 tabs

  6. A Perspective on Equipment Design for Fusion Remote Handling

    International Nuclear Information System (INIS)

    Mills, S.; Haist, B.; Hamilton, D.

    2006-01-01

    For 8 years, JET remote operations have become more capable and confident. Many tasks have been successfully completed, even those never intended to be remote maintenance activities. The general approach to the provision of remote handling equipment at JET has been the preferred use of commercially-off-the-shelf equipment. In the areas of electrical, electronic, software and control this approach has been generally achievable. However, in the area of mechanical equipment it has been more difficult. In particular the RH tooling has been almost entirely bespoke as its requirements are highly sensitive to the design of the JET component being handled and there are many design variations. Hence, JET has required the design and manufacture of over 700 types of bespoke RH equipment. This paper will discuss the experience of introducing and developing remote handling mechanical equipment for JET. The paper will cover the relationship between the remote handling equipment and the JET component design and the potential for improving the design function. A major lesson from the introduction of remote handling to JET has been demonstration of the very close interdependency of the design of JET components with design of remote handling tooling. The JET remote handling manual was originally introduced as the vehicle to ensure remote handling compatibility by the introduction of standards. Experience has shown that in general the remote handling manual approach has been insufficient. Future fusion machines will be much more complex than JET and will demand even greater remote handling compatibility. This paper will discuss possible methods for improving this process. Equipment operating in a high radiation environment must be dependable It may spend part of its time in areas that would be extremely difficult to recover from in the case of failure. The equipment may also have a high duty cycle to minimise shutdown times and probably cannot be manually inspected on a frequent

  7. A preliminary conceptual design study for Korean fusion DEMO reactor

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Keeman, E-mail: kkeeman@nfri.re.kr [National Fusion Research Institute, 169-148 Gwahak-ro, Daejeon 305-806 (Korea, Republic of); Kim, Hyoung Chan; Oh, Sangjun; Lee, Young Seok; Yeom, Jun Ho; Im, Kihak; Lee, Gyung-Su [National Fusion Research Institute, 169-148 Gwahak-ro, Daejeon 305-806 (Korea, Republic of); Neilson, George; Kessel, Charles; Brown, Thomas; Titus, Peter [Princeton Plasma Physics Laboratory, Princeton University, Princeton, NJ 08543 (United States)

    2013-10-15

    Highlights: ► Perform a preliminary conceptual study for a steady-state Korean DEMO reactor. ► Present design guidelines and requirements of Korean DEMO reactor. ► Present a preliminary design of TF (toroidal field) and CS (central solenoid) magnet. ► Present a preliminary result of the radial build scheme of Korean DEMO reactor. -- Abstract: As the ITER is being constructed, there is a growing anticipation for an earlier realization of fusion energy, so called fast-track approach. Korean strategy for fusion energy can be regarded as a fast-track approach and one special concept discussed in this paper is a two-stage development plan. At first, a steady-state Korean DEMO Reactor (K-DEMO) is designed not only to demonstrate a net electricity generation and a self-sustained tritium cycle, but also to be used as a component test facility. Then, at its second stage, a major upgrade is carried out by replacing in-vessel components in order to show a net electric generation on the order of 300 MWe and the competitiveness in cost of electricity (COE). The major radius is designed to be just below 6.5 m, considering practical engineering feasibilities. By using high performance Nb{sub 3}Sn-based superconducting cable currently available, high magnetic field at the plasma center above 8 T can be achieved. A design concept for TF magnets and radial builds for the K-DEMO considering a vertical maintenance scheme, are presented together with preliminary design parameters.

  8. Proposed design criteria for a fusion facility electrical ground system

    International Nuclear Information System (INIS)

    Armellino, C.A.

    1983-01-01

    Ground grid design considerations for a nuclear fusion reactor facility are no different than any other facility in that the basis for design must be safety first and foremost. Unlike a conventional industrial facility the available fault energy comes not only from the utility source and in-house rotating machinery, but also from energy storage capacitor banks, collapsing magnetic fields and D.C. transmission lines. It is not inconceivable for a fault condition occurrence where all available energy can be discharged. The ground grid must adequately shunt this sudden energy discharge in a way that personnel will not be exposed by step and/or touch to hazardous energy levels that are in excess of maximum tolerable levels for humans. Fault energy discharge rate is a function of the ground grid surge impedance characteristic. Closed loop paths must be avoided in the ground grid design so that during energy discharge no stray magnetic fields or large voltage potentials between remote points can be created by circulating currents. Single point connection of equipment to the ground grid will afford protection to personnel and sensitive equipment by reducing the probability of circulating currents. The overall ground grid system design is best illustrated as a wagon wheel concept with the fusion machine at the center. Radial branches or spokes reach out to the perimeter limits designated by step-and-touch high risk areas based on soil resistivity criteria considerations. Conventional methods for the design of a ground grid with all of its radial branches are still pertinent. The center of the grid could include a deep well single ground rod element the length of which is at least equivalent to the radius of an imaginary sphere that enshrouds the immediate machine area. Special facilities such as screen rooms or other shielded areas are part of the ground grid system by way of connection to radial branches

  9. Introduction to Nuclear Fusion Power and the Design of Fusion Reactors. An Issue-Oriented Module.

    Science.gov (United States)

    Fillo, J. A.

    This three-part module focuses on the principles of nuclear fusion and on the likely nature and components of a controlled-fusion power reactor. The physical conditions for a net energy release from fusion and two approaches (magnetic and inertial confinement) which are being developed to achieve this goal are described. Safety issues associated…

  10. Role of protein disulfide isomerase and other thiol-reactive proteins in HIV-1 envelope protein-mediated fusion

    International Nuclear Information System (INIS)

    Ou Wu; Silver, Jonathan

    2006-01-01

    Cell-surface protein disulfide isomerase (PDI) has been proposed to promote disulfide bond rearrangements in HIV-1 envelope protein (Env) that accompany Env-mediated fusion. We evaluated the role of PDI in ways that have not been previously tested by downregulating PDI with siRNA and by overexpressing wild-type or variant forms of PDI in transiently and stably transfected cells. These manipulations, as well as treatment with anti-PDI antibodies, had only small effects on infection or cell fusion mediated by NL4-3 or AD8 strains of HIV-1. However, the cell-surface thiol-reactive reagent 5, 5'-dithiobis(2-nitrobenzoic acid) (DTNB) had a much stronger inhibitory effect in our system, suggesting that cell-surface thiol-containing molecules other than PDI, acting alone or in concert, have a greater effect than PDI on HIV-1 Env-mediated fusion. We evaluated one such candidate, thioredoxin, a PDI family member reported to reduce a labile disulfide bond in CD4. We found that the ability of thioredoxin to reduce the disulfide bond in CD4 is enhanced in the presence of HIV-1 Env gp120 and that thioredoxin also reduces disulfide bonds in gp120 directly in the absence of CD4. We discuss the implications of these observations for identification of molecules involved in disulfide rearrangements in Env during fusion

  11. Automation of fusion first wall design using artificial intelligence technique

    International Nuclear Information System (INIS)

    Yoshimura, Shinobu; Yagawa, Genki; Mochizuki, Yoshihiko

    1990-01-01

    This paper describes the application of artificial intelligence techniques to a design automation of the fusion first wall to be operated in the complex environment where huge electromagnetic and thermal loading as well as heavy neutron irradiation occur. As a basic strategy of designing structure shape considering many coupled phenomena, an ordinary design procedure based on the generate and test strategy is adopted because of its simplicity and broad applicability. To automate the design procedure with maintaining its flexibility, extensibility and efficiency, artificial intelligence techniques are utilized in the following. An object-oriented knowledge representation technique is adopted to store knowledge modules, that is, objects, related to the first wall design, while a data-flow processing technique is utilized as an inference mechanism among the knowledge modules. These techniques realize the flexibility and extensibility of the system. Moreover, as an efficient design modification mechanism, which is essential in a design process, an empirical approach based on experts' empirical knowledge and a mathematical approach based on a kind of numerical sensitivity analysis are introduced. The developed system is applied to a simple example of the design of a two-dimensional model of the first wall with a cooling channel, and its fundamental performance is clearly demonstrated. (author)

  12. Recent developments in the design of conceptual fusion reactors

    International Nuclear Information System (INIS)

    Ribe, F.L.

    1977-01-01

    Since the first round of conceptual fusion reactor designs in 1973 - 1974, there has been considerable progress in design improvement. Two recent tokamak designs of the Wisconsin and Culham groups, with increased plasma beta and wall loading (power density), lead to more compact reactors with easier maintenance. The Reference Theta-Pinch Reactor has undergone considerable upgrading in the design of the first wall insulator and blanket. In addition, a conceptual homopolar energy storage and transfer system has been designed. In the case of the mirror reactor, there are design changes toward improved modular construction and ease of handling, as well as improved direct converters. Conceptual designs of toroidal-multiple-mirror, liner-compression, and reverse-field pinch reactors are also discussed. A design is presented of a toroidal multiple-mirror reactor that combines the advantages of steady-state operation and high-aspect ratio. The liner-compression reactor eliminates a major problem of radiation damage by using a liquid-metal first wall that also serves as a neutron-thermalizing blanket. The reverse-field pinch reactor operates at higher beta, larger current density and larger aspect ratio than a tokamak reactor. These properties allow the possibility of ignition by ohmic heating alone and greater ease of maintenance

  13. Human antibody recognition of antigenic site IV on Pneumovirus fusion proteins.

    Science.gov (United States)

    Mousa, Jarrod J; Binshtein, Elad; Human, Stacey; Fong, Rachel H; Alvarado, Gabriela; Doranz, Benjamin J; Moore, Martin L; Ohi, Melanie D; Crowe, James E

    2018-02-01

    Respiratory syncytial virus (RSV) is a major human pathogen that infects the majority of children by two years of age. The RSV fusion (F) protein is a primary target of human antibodies, and it has several antigenic regions capable of inducing neutralizing antibodies. Antigenic site IV is preserved in both the pre-fusion and post-fusion conformations of RSV F. Antibodies to antigenic site IV have been described that bind and neutralize both RSV and human metapneumovirus (hMPV). To explore the diversity of binding modes at antigenic site IV, we generated a panel of four new human monoclonal antibodies (mAbs) and competition-binding suggested the mAbs bind at antigenic site IV. Mutagenesis experiments revealed that binding and neutralization of two mAbs (3M3 and 6F18) depended on arginine (R) residue R429. We discovered two R429-independent mAbs (17E10 and 2N6) at this site that neutralized an RSV R429A mutant strain, and one of these mAbs (17E10) neutralized both RSV and hMPV. To determine the mechanism of cross-reactivity, we performed competition-binding, recombinant protein mutagenesis, peptide binding, and electron microscopy experiments. It was determined that the human cross-reactive mAb 17E10 binds to RSV F with a binding pose similar to 101F, which may be indicative of cross-reactivity with hMPV F. The data presented provide new concepts in RSV immune recognition and vaccine design, as we describe the novel idea that binding pose may influence mAb cross-reactivity between RSV and hMPV. Characterization of the site IV epitope bound by human antibodies may inform the design of a pan-Pneumovirus vaccine.

  14. Conservation of a proteinase cleavage site between an insect retrovirus (gypsy) Env protein and a baculovirus envelope fusion protein

    International Nuclear Information System (INIS)

    Pearson, Margot N.; Rohrmann, George F.

    2004-01-01

    The predicted Env protein of insect retroviruses (errantiviruses) is related to the envelope fusion protein of a major division of the Baculoviridae. The highest degree of homology is found in a region that contains a furin cleavage site in the baculovirus proteins and an adjacent sequence that has the properties of a fusion peptide. In this investigation, the homologous region in the Env protein of the gypsy retrovirus of Drosophila melanogaster (DmegypV) was investigated. Alteration of the predicted DmegypV Env proteinase cleavage site from RIAR to AIAR significantly reduced cleavage of Env in both Spodoptera frugiperda (Sf-9) and D. melanogaster (S2) cell lines. When the predicted DmegypV Env cleavage site RIAR was substituted for the cleavage sequence RRKR in the Lymantria dispar nucleopolyhedrovirus fusion protein (LD130) sequence, cleavage of the hybrid LD130 molecules still occurred, although at a reduced level. The conserved 21-amino acid sequence just downstream of the cleavage site, which is thought to be the fusion peptide in LD130, was also characterized. When this sequence from DmegypV Env was substituted for the homologous sequence in LD130, cleavage still occurred, but no fusion was observed in either cell type. In addition, although a DmegypV-Env-green fluorescent protein construct localized to cell membranes, no cell fusion was observed

  15. Design study of electrostatically plugged cusp fusion reactor

    International Nuclear Information System (INIS)

    Dolan, T.J.

    1976-01-01

    This study concentrates on the following aspects of an electrostatically plugged cusp reactor that will be different from other fusion reactor designs: the coil geometry and structural supports, high voltage electrodes, plasma parameters, power balance, and operating cycle. Assuming the electron density distribution in the anodes to have a characteristic width of two electron Larmor radii, which is consistent with present experimental results, the theory predicts that a device with a magnetic field strength, B = 8 T sustained solely by electron beam injection at 300 kV will have a power gain ratio, Q, of about 5. A toroidal multipole cusp configuration with six cusps was selected for the present design, based on a study of the ratio of plasma volume to coil volume. Coil forces are sustained by cryogenic trusses between like coils, fiberglass compression columns, and room temperature hoops. Radiation collimators in front of the high voltage electrodes greatly reduce the radiation impinging on the cathodes, helping to avoid breakdown and to prolong insulator life. The operating cycle consists of a startup period of about 20 s, followed by a fusion burn period lasting about 200 s (limited by impurity buildup) and a 20-s flushing period

  16. Production of N-acetyl-D-neuraminic acid using two sequential enzymes overexpressed as double-tagged fusion proteins

    Directory of Open Access Journals (Sweden)

    Cheng Chung-Hsien

    2009-07-01

    Full Text Available Abstract Background Two sequential enzymes in the production of sialic acids, N-acetyl-D-glucosamine 2-epimerase (GlcNAc 2-epimerase and N-acetyl-D-neuraminic acid aldolase (Neu5Ac aldolase, were overexpressed as double-tagged gene fusions. Both were tagged with glutathione S-transferase (GST at the N-terminus, but at the C-terminus, one was tagged with five contiguous aspartate residues (5D, and the other with five contiguous arginine residues (5R. Results Both fusion proteins were overexpressed in Escherichia coli and retained enzymatic activity. The fusions were designed so their surfaces were charged under enzyme reaction conditions, which allowed isolation and immobilization in a single step, through a simple capture with either an anionic or a cationic exchanger (Sepharose Q or Sepharose SP that electrostatically bound the 5D or 5R tag. The introduction of double tags only marginally altered the affinity of the enzymes for their substrates, and the double-tagged proteins were enzymatically active in both soluble and immobilized forms. Combined use of the fusion proteins led to the production of N-acetyl-D-neuraminic acid (Neu5Ac from N-acetyl-D-glucosamine (GlcNAc. Conclusion Double-tagged gene fusions were overexpressed to yield two enzymes that perform sequential steps in sialic acid synthesis. The proteins were easily immobilized via ionic tags onto ionic exchange resins and could thus be purified by direct capture from crude protein extracts. The immobilized, double-tagged proteins were effective for one-pot enzymatic production of sialic acid.

  17. Single chain Fc-dimer-human growth hormone fusion protein for improved drug delivery.

    Science.gov (United States)

    Zhou, Li; Wang, Hsuan-Yao; Tong, Shanshan; Okamoto, Curtis T; Shen, Wei-Chiang; Zaro, Jennica L

    2017-02-01

    Fc fusion protein technology has been successfully used to generate long-acting forms of several protein therapeutics. In this study, a novel Fc-based drug carrier, single chain Fc-dimer (sc(Fc) 2 ), was designed to contain two Fc domains recombinantly linked via a flexible linker. Since the Fc dimeric structure is maintained through the flexible linker, the hinge region was omitted to further stabilize it against proteolysis and reduce FcγR-related effector functions. The resultant sc(Fc) 2 candidate preserved the neonatal Fc receptor (FcRn) binding. sc(Fc) 2 -mediated delivery was then evaluated using a therapeutic protein with a short plasma half-life, human growth hormone (hGH), as the protein drug cargo. This novel carrier protein showed a prolonged in vivo half-life and increased hGH-mediated bioactivity compared to the traditional Fc-based drug carrier. sc(Fc) 2 technology has the potential to greatly advance and expand the use of Fc-technology for improving the pharmacokinetics and bioactivity of protein therapeutics. Copyright © 2016 Elsevier Ltd. All rights reserved.

  18. Structural Transition and Antibody Binding of EBOV GP and ZIKV E Proteins from Pre-Fusion to Fusion-Initiation State

    Directory of Open Access Journals (Sweden)

    Anna Lappala

    2018-05-01

    Full Text Available Membrane fusion proteins are responsible for viral entry into host cells—a crucial first step in viral infection. These proteins undergo large conformational changes from pre-fusion to fusion-initiation structures, and, despite differences in viral genomes and disease etiology, many fusion proteins are arranged as trimers. Structural information for both pre-fusion and fusion-initiation states is critical for understanding virus neutralization by the host immune system. In the case of Ebola virus glycoprotein (EBOV GP and Zika virus envelope protein (ZIKV E, pre-fusion state structures have been identified experimentally, but only partial structures of fusion-initiation states have been described. While the fusion-initiation structure is in an energetically unfavorable state that is difficult to solve experimentally, the existing structural information combined with computational approaches enabled the modeling of fusion-initiation state structures of both proteins. These structural models provide an improved understanding of four different neutralizing antibodies in the prevention of viral host entry.

  19. Safety considerations in the design of the fusion engineering device

    International Nuclear Information System (INIS)

    Barrett, R.J.

    1983-01-01

    Safety considerations play a significant role in the design of a near-term Fusion Engineering Device (FED). For the safety of the general public and the plant workers, the radiation environment caused by the reacting plasma and the potential release of tritium fuel are the dominant considerations. The U.S. Department of Energy (DOE) regulations and guidelines for radiation protection have been reviewed and are being applied to the device design. Direct radiation protection is provided by the device shield and the reactor building walls. Radiation from the activated device components and the tritium fuel is to be controlled with shielding, contamination control, and ventilation. The potential release of tritium from the plant has influenced the selection of reactor building and plant designs and specifications. The safety of the plant workers is affected primarily by the radiation from the activated device components and from plasma chamber debris. The highly activated device components make it necessary to design many of the maintenance activities in the reactor building for totally remote operation. The hot cell facility has evolved as a totally remote maintenance facility due to the high radiation levels of the device components. Safety considerations have had substantial impacts on the design of FED. Several examples of safety-related design impacts are discussed in the paper. Feasible solutions have been identified for all outstanding safety-related items, and additional optimization of these solutions is anticipated in future design studies

  20. Towards a strategy of reliable fusion first-wall design

    International Nuclear Information System (INIS)

    Schultz, J.H.

    1981-05-01

    Fusion first walls are subject to a large number of possible failure mechanisms, including erosion due to sputtering, arcing, blistering and vaporization and crack growth due to thermal and magnetic stresses. Each of these failure mechanisms is poorly characterized and has the potential of being severe. A strategy for designing reliably in the face of great uncertainty is discussed. Topological features beneficial to reactor availability are identified. The integration of limiter pumping with rf wave launching is discussed, as a means of simplifying reactor design. The concept of a sewer limiter is introduced, as a possible long-life limiter topology. The concept of flexible armor is discussed, as a means of extending maximum life

  1. A conceptual design of LIB fusion reactor: UTLIF(2)

    International Nuclear Information System (INIS)

    Madarame, Haruki; Kondo, Shunsuke; Iwata, Shuichi; Oka, Yoshiaki; Miya, Kenzo.

    1984-01-01

    UTLIF(2) is a conceptual design study on a light ion beam driven fusion reactor based on a concept of rod-bundle blanket. Survivability and maintainability of the first wall and the blanket are regarded as of major importance in the design. The blanket rod is composed of a thick tube which has enough stiffness, a thin wrapping wall which receives high heat flux, and liquid lithium which breeds tritium and removes generated heat. The rod can be pulled out from the outside of the reactor vessel, hence the replacement is very easy. Nuclear and thermal analysis have been made and the performance of the reactor has been shown to be satisfactory. (author)

  2. Conceptual design study of fusion experimental reactor (FY86 FER)

    International Nuclear Information System (INIS)

    Seki, Yasushi; Iida, Hiromasa; Honda, Tsutomu.

    1987-08-01

    This report describes the study on safety for FER(Fusion Experimental Reactor) which has been designed as a next step machine to the JT-60. Though the final purpose of this study is to have an image of design base accident, maximum credible accident and to assess their risk or probability, etc., as FER plant system, the emphasis of this years study is placed on fuel-gas circulation system where the tritium inventory is maximum. This report consists of two chapters. The first chapter of this report summaries the FER system and describes FMEA(Failure Mode and Effect Analysis) and related accident progression sequence for FER plant system as a whole. The second chapter of this report is focused on fuel-gas circulation system including the purification, isotope separation system and storage system. Here, probability of risk is assessed by the probabilistic risk analysis (PRA) procedure based on FMEA, ETA and FTA. (author)

  3. Fusion peptides from oncogenic chimeric proteins as putative specific biomarkers of cancer.

    Science.gov (United States)

    Conlon, Kevin P; Basrur, Venkatesha; Rolland, Delphine; Wolfe, Thomas; Nesvizhskii, Alexey I; MacCoss, Michael J; Lim, Megan S; Elenitoba-Johnson, Kojo S J

    2013-10-01

    Chromosomal translocations encoding chimeric fusion proteins constitute one of the most common mechanisms underlying oncogenic transformation in human cancer. Fusion peptides resulting from such oncogenic chimeric fusions, though unique to specific cancer subtypes, are unexplored as cancer biomarkers. Here we show, using an approach termed fusion peptide multiple reaction monitoring mass spectrometry, the direct identification of different cancer-specific fusion peptides arising from protein chimeras that are generated from the juxtaposition of heterologous genes fused by recurrent chromosomal translocations. Using fusion peptide multiple reaction monitoring mass spectrometry in a clinically relevant scenario, we demonstrate the specific, sensitive, and unambiguous detection of a specific diagnostic fusion peptide in clinical samples of anaplastic large cell lymphoma, but not in a diverse array of benign lymph nodes or other forms of primary malignant lymphomas and cancer-derived cell lines. Our studies highlight the utility of fusion peptides as cancer biomarkers and carry broad implications for the use of protein biomarkers in cancer detection and monitoring.

  4. Conceptual design of China fusion power plant FDS-II

    International Nuclear Information System (INIS)

    Wu, Y.; Liu, S.; Chen, H.

    2007-01-01

    As one of the series of fusion system design concepts developed by the FDS Team of China, FDS-II is designated to exploit and evaluate potential attractiveness of fusion energy application for the generation of electricity on the basis of conservatively advanced plasma parameters, which can be limitedly extrapolated from the successful operation of ITER. The principle of the blanket design is established in both the feasibility and potential attractiveness of technology to meet the requirement for tritium self-sufficiency, safety margin, operation economy and environment protection etc. The plasma physics and engineering parameters of FDS-II are selected on the basis of the progress in recent experiments and associated theoretical studies of magnetic confinement fusion plasma with a fusion power of 2∝3 GW. The neutron wall load of 2∝3 MW/m 2 and the surface heat flux of 0.5∝1 MW/m 2 are considered for high effective power conversion. The ''multi-modules'' scenario is adopted in the FDS-II blanket design to reduce thermal stress and electromagnetic forces under plasma disruption, with liquid metal lithium lead (LiPb) as tritium breeder, the Reduced Activation Ferritic/Martensitic (RAFM) steel as structural material. Two options of specific liquid LiPb blanket concepts have been proposed, named the Dual-cooled Lithium Lead (DLL) breeder blanket and the Quasi-Static Lithium Lead (SLL) breeder blanket. The DLL blanket is a dual-cooled LiPb breeder system with helium gas to cool the first wall and main structure and LiPb eutectic to be self-cooled. The flow channel inserts (FCIs), e.g. SiCf/SiC composites, are designed as the thermal and electrical insulators inside the LiPb flow channels to reduce the magnetohydrodynamic (MHD) pressure drop and to allow the coolant LiPb outlet temperature up to 700 C for high thermal efficiency. The SLL blanket is another option of the FDS-II blanket with the technology developed relatively easily. To avoid or mitigate the

  5. The TIP30 protein complex, arachidonic acid and coenzyme A are required for vesicle membrane fusion.

    Directory of Open Access Journals (Sweden)

    Chengliang Zhang

    Full Text Available Efficient membrane fusion has been successfully mimicked in vitro using artificial membranes and a number of cellular proteins that are currently known to participate in membrane fusion. However, these proteins are not sufficient to promote efficient fusion between biological membranes, indicating that critical fusogenic factors remain unidentified. We have recently identified a TIP30 protein complex containing TIP30, acyl-CoA synthetase long-chain family member 4 (ACSL4 and Endophilin B1 (Endo B1 that promotes the fusion of endocytic vesicles with Rab5a vesicles, which transport endosomal acidification enzymes vacuolar (H⁺-ATPases (V-ATPases to the early endosomes in vivo. Here, we demonstrate that the TIP30 protein complex facilitates the fusion of endocytic vesicles with Rab5a vesicles in vitro. Fusion of the two vesicles also depends on arachidonic acid, coenzyme A and the synthesis of arachidonyl-CoA by ACSL4. Moreover, the TIP30 complex is able to transfer arachidonyl groups onto phosphatidic acid (PA, producing a new lipid species that is capable of inducing close contact between membranes. Together, our data suggest that the TIP30 complex facilitates biological membrane fusion through modification of PA on membranes.

  6. Hefei experimental hybrid fusion-fission reactor conceptual design

    International Nuclear Information System (INIS)

    Qiu Lijian; Luan Guishi; Xu Qiang

    1992-03-01

    A new concept of hybrid reactor is introduced. It uses JET-like(Joint European Tokamak) device worked at sub-breakeven conditions, as a source of high energy neutrons to induce a blanket fission of depleted uranium. The solid breeding material and helium cooling technique are also used. It can produce 100 kg of 239 Pu per year by partial fission suppressed. The energy self-sustained of the fusion core is not necessary. Plasma temperature is maintained by external 20 MW ICRF (ion cyclotron resonance frequency) and 10 MW ECRF (electron cyclotron resonance frequency) heating. A steady state plasma current at 1.5 Ma is driven by 10 MW LHCD (lower hybrid current driven). Plasma density will be kept by pellet injection. ICRF can produce a high energy tail in ion distribution function and lead to significant enhancement of D-T reaction rate by 2 ∼ 5 times so that the neutron source strength reaches to the level of 1 x 10 19 n/s. This system is a passive system. It's power density is 10 W/cm 3 and the wall loading is 0.6 W/cm 2 that is the lower limitation of fusion and fission technology. From the calculation of neutrons it could always be in sub-critical and has intrinsic safety. The radiation damage and neutron flux distribution on the first wall are also analyzed. According to the conceptual design the application of this type hybrid reactor earlier is feasible

  7. Rapamycin-induced oligomer formation system of FRB-FKBP fusion proteins.

    Science.gov (United States)

    Inobe, Tomonao; Nukina, Nobuyuki

    2016-07-01

    Most proteins form larger protein complexes and perform multiple functions in the cell. Thus, artificial regulation of protein complex formation controls the cellular functions that involve protein complexes. Although several artificial dimerization systems have already been used for numerous applications in biomedical research, cellular protein complexes form not only simple dimers but also larger oligomers. In this study, we showed that fusion proteins comprising the induced heterodimer formation proteins FRB and FKBP formed various oligomers upon addition of rapamycin. By adjusting the configuration of fusion proteins, we succeeded in generating an inducible tetramer formation system. Proteins of interest also formed tetramers by fusing to the inducible tetramer formation system, which exhibits its utility in a broad range of biological applications. Copyright © 2015 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  8. Polyclonal and monoclonal antibodies specific for the six-helix bundle of the human respiratory syncytial virus fusion glycoprotein as probes of the protein post-fusion conformation

    International Nuclear Information System (INIS)

    Palomo, Concepción; Mas, Vicente; Vázquez, Mónica; Cano, Olga; Luque, Daniel; Terrón, María C.; Calder, Lesley J.; Melero, José A.

    2014-01-01

    Human respiratory syncytial virus (hRSV) has two major surface glycoproteins (G and F) anchored in the lipid envelope. Membrane fusion promoted by hRSV F occurs via refolding from a pre-fusion form to a highly stable post-fusion state involving large conformational changes of the F trimer. One of these changes results in assembly of two heptad repeat sequences (HRA and HRB) into a six-helix bundle (6HB) motif. To assist in distinguishing pre- and post-fusion conformations of hRSV F , we have prepared polyclonal (α-6HB) and monoclonal (R145) rabbit antibodies specific for the 6HB. Among other applications, these antibodies were used to explore the requirements of 6HB formation by isolated protein segments or peptides and by truncated mutants of the F protein. Site-directed mutagenesis and electron microscopy located the R145 epitope in the post-fusion hRSV F at a site distantly located from previously mapped epitopes, extending the repertoire of antibodies that can decorate the F molecule. - Highlights: • Antibodies specific for post-fusion respiratory syncytial virus fusion protein are described. • Polyclonal antibodies were obtained in rabbit inoculated with chimeric heptad repeats. • Antibody binding required assembly of a six-helix bundle in the post-fusion protein. • A monoclonal antibody with similar structural requirements is also described. • Binding of this antibody to the post-fusion protein was visualized by electron microscopy

  9. Polyclonal and monoclonal antibodies specific for the six-helix bundle of the human respiratory syncytial virus fusion glycoprotein as probes of the protein post-fusion conformation

    Energy Technology Data Exchange (ETDEWEB)

    Palomo, Concepción; Mas, Vicente; Vázquez, Mónica; Cano, Olga [Unidad de Biología Viral, Centro Nacional de Microbiología, Madrid (Spain); CIBER de Enfermedades Respiratorias, Instituto de Salud Carlos III, Majadahonda, 28220 Madrid (Spain); Luque, Daniel; Terrón, María C. [Unidad de Microscopía Electrónica y Confocal, Centro Nacional de Microbiología, Instituto de Salud Carlos III, Majadahonda, 28220 Madrid (Spain); Calder, Lesley J. [National Institute for Medical Research, MRC, Mill Hill, London NW7 1AA (United Kingdom); Melero, José A., E-mail: jmelero@isciii.es [Unidad de Biología Viral, Centro Nacional de Microbiología, Madrid (Spain); CIBER de Enfermedades Respiratorias, Instituto de Salud Carlos III, Majadahonda, 28220 Madrid (Spain)

    2014-07-15

    Human respiratory syncytial virus (hRSV) has two major surface glycoproteins (G and F) anchored in the lipid envelope. Membrane fusion promoted by hRSV{sub F} occurs via refolding from a pre-fusion form to a highly stable post-fusion state involving large conformational changes of the F trimer. One of these changes results in assembly of two heptad repeat sequences (HRA and HRB) into a six-helix bundle (6HB) motif. To assist in distinguishing pre- and post-fusion conformations of hRSV{sub F}, we have prepared polyclonal (α-6HB) and monoclonal (R145) rabbit antibodies specific for the 6HB. Among other applications, these antibodies were used to explore the requirements of 6HB formation by isolated protein segments or peptides and by truncated mutants of the F protein. Site-directed mutagenesis and electron microscopy located the R145 epitope in the post-fusion hRSV{sub F} at a site distantly located from previously mapped epitopes, extending the repertoire of antibodies that can decorate the F molecule. - Highlights: • Antibodies specific for post-fusion respiratory syncytial virus fusion protein are described. • Polyclonal antibodies were obtained in rabbit inoculated with chimeric heptad repeats. • Antibody binding required assembly of a six-helix bundle in the post-fusion protein. • A monoclonal antibody with similar structural requirements is also described. • Binding of this antibody to the post-fusion protein was visualized by electron microscopy.

  10. Design of a hyperstable 60-subunit protein icosahedron

    Science.gov (United States)

    Hsia, Yang; Bale, Jacob B.; Gonen, Shane; Shi, Dan; Sheffler, William; Fong, Kimberly K.; Nattermann, Una; Xu, Chunfu; Huang, Po-Ssu; Ravichandran, Rashmi; Yi, Sue; Davis, Trisha N.; Gonen, Tamir; King, Neil P.; Baker, David

    2016-07-01

    The icosahedron is the largest of the Platonic solids, and icosahedral protein structures are widely used in biological systems for packaging and transport. There has been considerable interest in repurposing such structures for applications ranging from targeted delivery to multivalent immunogen presentation. The ability to design proteins that self-assemble into precisely specified, highly ordered icosahedral structures would open the door to a new generation of protein containers with properties custom-tailored to specific applications. Here we describe the computational design of a 25-nanometre icosahedral nanocage that self-assembles from trimeric protein building blocks. The designed protein was produced in Escherichia coli, and found by electron microscopy to assemble into a homogenous population of icosahedral particles nearly identical to the design model. The particles are stable in 6.7 molar guanidine hydrochloride at up to 80 degrees Celsius, and undergo extremely abrupt, but reversible, disassembly between 2 molar and 2.25 molar guanidinium thiocyanate. The icosahedron is robust to genetic fusions: one or two copies of green fluorescent protein (GFP) can be fused to each of the 60 subunits to create highly fluorescent ‘standard candles’ for use in light microscopy, and a designed protein pentamer can be placed in the centre of each of the 20 pentameric faces to modulate the size of the entrance/exit channels of the cage. Such robust and customizable nanocages should have considerable utility in targeted drug delivery, vaccine design and synthetic biology.

  11. Mycobacterium tuberculosis HspX/EsxS Fusion Protein: Gene Cloning, Protein Expression, and Purification in Escherichia coli.

    Science.gov (United States)

    Khademi, Farzad; Yousefi-Avarvand, Arshid; Derakhshan, Mohammad; Meshkat, Zahra; Tafaghodi, Mohsen; Ghazvini, Kiarash; Aryan, Ehsan; Sankian, Mojtaba

    2017-10-01

    The purpose of this study was to clone, express, and purify a novel multidomain fusion protein of Micobacterium tuberculosis (Mtb) in a prokaryotic system. An hspX/esxS gene construct was synthesized and ligated into a pGH plasmid, E. coli TOP10 cells were transformed, and the vector was purified. The vector containing the construct and pET-21b (+) plasmid were digested with the same enzymes and the construct was ligated into pET-21b (+). The accuracy of cloning was confirmed by colony PCR and sequencing. E. coli BL21 cells were transformed with the pET-21b (+)/hspX/esxS expression vector and protein expression was evaluated. Finally, the expressed fusion protein was purified on a Ni-IDA column and verified by SDS-PAGE and western blotting. The hspX/esxS gene construct was inserted into pET-21b (+) and recombinant protein expression was induced with IPTG in E. coli BL21 cells. Various concentrations of IPTG were tested to determine the optimum concentration for expression induction. The recombinant protein was expressed in insoluble inclusion bodies. Three molar guanidine HCl was used to solubilize the insoluble protein. An HspX/EsxS Mtb fusion protein was expressed in E. coli and the recombinant protein was purified. After immunological analysis, the HspX/EsxS fusion protein might be an anti-tuberculosis vaccine candidate in future clinical trial studies.

  12. Production of recombinant proteins in Escherichia coli tagged with the fusion protein CusF3H.

    Science.gov (United States)

    Vargas-Cortez, Teresa; Morones-Ramirez, Jose Ruben; Balderas-Renteria, Isaias; Zarate, Xristo

    2017-04-01

    Recombinant protein expression in the bacterium Escherichia coli still is the number one choice for large-scale protein production. Nevertheless, many complications can arise using this microorganism, such as low yields, the formation of inclusion bodies, and the requirement for difficult purification steps. Most of these problems can be solved with the use of fusion proteins. Here, the use of the metal-binding protein CusF3H+ is described as a new fusion protein for recombinant protein expression and purification in E. coli. We have previously shown that CusF produces large amounts of soluble protein, with low levels of formation of inclusion bodies, and that proteins can be purified using IMAC resins charged with Cu(II) ions. CusF3H+ is an enhanced variant of CusF, formed by the addition of three histidine residues at the N-terminus. These residues then can bind Ni(II) ions allowing improved purity after affinity chromatography. Expression and purification of Green Fluorescent Protein tagged with CusF3H+ showed that the mutation did not alter the capacity of the fusion protein to increase protein expression, and purity improved considerably after affinity chromatography with immobilized nickel ions; high yields are obtained after tag-removal since CusF3H+ is a small protein of just 10 kDa. Furthermore, the results of experiments involving expression of tagged proteins having medium to large molecular weights indicate that the presence of the CusF3H+ tag improves protein solubility, as compared to a His-tag. We therefore endorse CusF3H+ as a useful alternative fusion protein/affinity tag for production of recombinant proteins in E. coli. Copyright © 2017 Elsevier Inc. All rights reserved.

  13. A fungal biofilm reactor based on metal structured packing improves the quality of a Gla::GFP fusion protein produced by Aspergillus oryzae.

    Science.gov (United States)

    Zune, Q; Delepierre, A; Gofflot, S; Bauwens, J; Twizere, J C; Punt, P J; Francis, F; Toye, D; Bawin, T; Delvigne, F

    2015-08-01

    Fungal biofilm is known to promote the excretion of secondary metabolites in accordance with solid-state-related physiological mechanisms. This work is based on the comparative analysis of classical submerged fermentation with a fungal biofilm reactor for the production of a Gla::green fluorescent protein (GFP) fusion protein by Aspergillus oryzae. The biofilm reactor comprises a metal structured packing allowing the attachment of the fungal biomass. Since the production of the target protein is under the control of the promoter glaB, specifically induced in solid-state fermentation, the biofilm mode of culture is expected to enhance the global productivity. Although production of the target protein was enhanced by using the biofilm mode of culture, we also found that fusion protein production is also significant when the submerged mode of culture is used. This result is related to high shear stress leading to biomass autolysis and leakage of intracellular fusion protein into the extracellular medium. Moreover, 2-D gel electrophoresis highlights the preservation of fusion protein integrity produced in biofilm conditions. Two fungal biofilm reactor designs were then investigated further, i.e. with full immersion of the packing or with medium recirculation on the packing, and the scale-up potentialities were evaluated. In this context, it has been shown that full immersion of the metal packing in the liquid medium during cultivation allows for a uniform colonization of the packing by the fungal biomass and leads to a better quality of the fusion protein.

  14. Optimizing HIV-1 protease production in Escherichia coli as fusion protein

    Directory of Open Access Journals (Sweden)

    Piubelli Luciano

    2011-06-01

    Full Text Available Abstract Background Human immunodeficiency virus (HIV is the etiological agent in AIDS and related diseases. The aspartyl protease encoded by the 5' portion of the pol gene is responsible for proteolytic processing of the gag-pol polyprotein precursor to yield the mature capsid protein and the reverse transcriptase and integrase enzymes. The HIV protease (HIV-1Pr is considered an attractive target for designing inhibitors which could be used to tackle AIDS and therefore it is still the object of a number of investigations. Results A recombinant human immunodeficiency virus type 1 protease (HIV-1Pr was overexpressed in Escherichia coli cells as a fusion protein with bacterial periplasmic protein dithiol oxidase (DsbA or glutathione S-transferase (GST, also containing a six-histidine tag sequence. Protein expression was optimized by designing a suitable HIV-1Pr cDNA (for E. coli expression and to avoid autoproteolysis and by screening six different E. coli strains and five growth media. The best expression yields were achieved in E. coli BL21-Codon Plus(DE3-RIL host and in TB or M9 medium to which 1% (w/v glucose was added to minimize basal expression. Among the different parameters assayed, the presence of a buffer system (based on phosphate salts and a growth temperature of 37°C after adding IPTG played the main role in enhancing protease expression (up to 10 mg of chimeric DsbA:HIV-1Pr/L fermentation broth. GST:HIVPr was in part (50% produced as soluble protein while the overexpressed DsbA:HIV-1Pr chimeric protein largely accumulated in inclusion bodies as unprocessed fusion protein. A simple refolding procedure was developed on HiTrap Chelating column that yielded a refolded DsbA:HIV-1Pr with a > 80% recovery. Finally, enterokinase digestion of resolubilized DsbA:HIV-1Pr gave more than 2 mg of HIV-1Pr per liter of fermentation broth with a purity ≤ 80%, while PreScission protease cleavage of soluble GST:HIVPr yielded ~ 0.15 mg of pure HIV-1

  15. Divertor conceptual designs for a fusion power plant

    International Nuclear Information System (INIS)

    Norajitra, P.; Ihli, T.; Janeschitz, G.; Abdel-Khalik, S.; Mazul, I.; Malang, S.

    2007-01-01

    The development of a divertor concept for post-ITER fusion power plants is deemed to be an urgent task to meet the EU Fast Track scenario. Developing a divertor is particularly challenging due to the wide range of requirements to be met including the high incident peak heat flux, the blanket design with which the divertor has to be integrated, sputtering erosion of the plasma-facing material caused by the incident a particles, radiation effects on the properties of structural materials, and efficient recovery and conversion of the divertor thermal power (∝15% of the total fusion thermal power) by maximizing the coolant operating temperature while minimizing the pumping power. In the course of the EU PPCS, three near-term (A, B and AB) and two advanced power plant models (C, D) were investigated. Model A utilizes a water-cooled lead-lithium (WCLL) blanket and a water-cooled divertor with a peak heat flux of 15 MW/m 2 . Model B uses a He-cooled ceramics/beryllium pebble bed (HCPB) blanket and a He-cooled divertor concept (10 MW/m 2 ). Model AB uses a He-cooled lithium-lead (HCLL) blanket and a He-cooled divertor concept (10 MW/m 2 ). Model C is based on a dual-coolant (DC) blanket (lead/lithium self-cooled bulk and He-cooled structures) and a He-cooled divertor (10 MW/m 2 ). Model D employs a self-cooled lead/lithium (SCLL) blanket and lead-lithiumcooled divertor (5 MW/m 2 ). The values in parenthesis correspond to the maximum peak heat fluxes required. It can be noted that the helium-cooled divertor is used in most of the EU plant models; it has also been proposed for the US ARIES-CS reactor study. Since 2002, it has been investigated extensively in Europe under the PPCS with the goal of reaching a maximum heat flux of at least 10 MW/m2. Work has covered many areas including conceptual design, analysis, material and fabrication issues, and experiments. Generally, the helium-cooled divertor is considered to be a suitable solution for fusion power plants, as it

  16. Fusion Engineering Device (FED) first wall/shield design

    International Nuclear Information System (INIS)

    Sager, P.H.; Fuller, G.; Cramer, B.; Davisson, J.; Haines, J.; Kirchner, J.

    1981-01-01

    The torus of the Fusion Engineering Device (FED) is comprised of the bulk shield and its associated spool lstructure and support system, the first wall water-cooled panel and armor systems, and the pumped limiter. The bulk shielding is provided by ten shield sectors that are installed in the spool structure in such a way as to permit extraction of the sectors through the openings between adjacent toroidal field coils with a direct radial movement. The first wall armor is installed on the inboard and top interior walls of these sectors, and the water-cooled panels are installed on the outboard interior walls and the pumped limiter in the bottom of the sectors. The overall design of the first wall and shield system is described in this paper

  17. Heavy ion fusion (HIF) impulse injector design, construction, and checkout

    Energy Technology Data Exchange (ETDEWEB)

    Wilson, M. J., LLNL

    1998-05-04

    The following report describes the design, construction, and checkout of a high-voltage (HV) impulser built for the heavy ion fusion (HIF) project. The purpose of this impulser is to provide an adjustable diode voltage source of sufficient quality and level to allow the optimization of beam transport and accelerator sections of HIF. An elegant, low-impedance, high-energy storage capacitor circuit has been selected for this application. A retrofit to the diode region has been included to provide additional beam stability and a controlled rise time. The critical part of this circuit that is common to all candidates is the impedance matching component. The following report provides a description of the implemented circuit, the basic circuit variables for wave shaping, component screening techniques, resulting operating parameters, diode modifications, operating considerations, and fault protection.

  18. Accelerator conceptual design of the international fusion materials irradiation facility

    Energy Technology Data Exchange (ETDEWEB)

    Sugimoto, M.; Kinsho, M. [Japan Atomic Energy Res. Inst., Tokai, Ibaraki (Japan). Intense Neutron Source Lab.; Jameson, R.A.; Blind, B. [Los Alamos National Lab., NM (United States); Teplyakov, V. [Institute for High Energy Physics, Moscow (Russian Federation); Berwald, D.; Bruhwiler, D.; Peakock, M.; Rathke, J. [Northrop Grumman Corp., Bethpage, NY (United States); Deitinghoff, H.; Klein, H.; Pozimski, Y.; Volk, K. [Johann Wolfgang Goethe Univ., Frankfurt (Germany). Inst. fur Angewandte Phys.; Ferdinand, R.; Lagniel, J.-M. [CEA Saclay LNS, Gif-sur-Yvette (France); Miyahara, A. [Teikyo Univ., Tokyo (Japan); Olivier, M. [CEA DSM, Saclay, Gif-sur-Yvette (France); Piechowiak, E. [Northrop Grumman Corp., Baltimore, MD (United States); Tanabe, Y. [Toshiba Corp., Tsurumi-ku, Yokohama (Japan)

    1998-10-01

    The accelerator system of the international fusion materials irradiation facility (IFMIF) provides the 250-mA, 40-MeV continuous-wave deuteron beam at one of the two lithium target stations. It consists of two identical linear accelerator modules, each of which independently delivers a 125-mA beam to the common footprint of 20 cm x 5 cm at the target surface. The accelerator module consists of an ion injector, a 175 MHz RFQ and eight DTL tanks, and rf power supply system. The requirements for the accelerator system and the design concept are described. The interface issues and operational considerations to attain the proposed availability are also discussed. (orig.) 8 refs.

  19. Accelerator conceptual design of the international fusion materials irradiation facility

    International Nuclear Information System (INIS)

    Sugimoto, M.; Kinsho, M.; Teplyakov, V.; Berwald, D.; Bruhwiler, D.; Peakock, M.; Rathke, J.; Deitinghoff, H.; Klein, H.; Pozimski, Y.; Volk, K.; Miyahara, A.; Olivier, M.; Piechowiak, E.; Tanabe, Y.

    1998-01-01

    The accelerator system of the international fusion materials irradiation facility (IFMIF) provides the 250-mA, 40-MeV continuous-wave deuteron beam at one of the two lithium target stations. It consists of two identical linear accelerator modules, each of which independently delivers a 125-mA beam to the common footprint of 20 cm x 5 cm at the target surface. The accelerator module consists of an ion injector, a 175 MHz RFQ and eight DTL tanks, and rf power supply system. The requirements for the accelerator system and the design concept are described. The interface issues and operational considerations to attain the proposed availability are also discussed. (orig.)

  20. Incorporation of albumin fusion proteins into fibrin clots in vitro and in vivo: comparison of different fusion motifs recognized by factor XIIIa

    Directory of Open Access Journals (Sweden)

    Sheffield William P

    2011-12-01

    Full Text Available Abstract Background The transglutaminase activated factor XIII (FXIIIa acts to strengthen pathological fibrin clots and to slow their dissolution, in part by crosslinking active α2-antiplasmin (α2AP to fibrin. We previously reported that a yeast-derived recombinant fusion protein comprising α2AP residues 13-42 linked to human serum albumin (HSA weakened in vitro clots but failed to become specifically incorporated into in vivo clots. In this study, our aims were to improve both the stability and clot localization of the HSA fusion protein by replacing α2AP residues 13-42 with shorter sequences recognized more effectively by FXIIIa. Results Expression plasmids were prepared encoding recombinant HSA with the following N-terminal 23 residue extensions: H6NQEQVSPLTLLAG4Y (designated XL1; H6DQMMLPWAVTLG4Y (XL2; H6WQHKIDLPYNGAG4Y (XL3; and their 17 residue non-His-tagged equivalents (XL4, XL5, and XL6. The HSA moiety of XL4- to XL6-HSA proteins was C-terminally His-tagged. All chimerae were efficiently secreted from transformed Pichia pastoris yeast except XL3-HSA, and following nickel chelate affinity purification were found to be intact by amino acid sequencing, as was an N-terminally His-tagged version of α2AP(13-42-HSA. Of the proteins tested, XL5-HSA was cross-linked to biotin pentylamine (BPA most rapidly by FXIIIa, and was the most effective competitor of α2AP crosslinking not only to BPA but also to plasma fibrin clots. In the mouse ferric chloride vena cava thrombosis model, radiolabeled XL5-HSA was retained in the clot to a greater extent than recombinant HSA. In the rabbit jugular vein stasis thrombosis model, XL5-HSA was also retained in the clot, in a urea-insensitive manner indicative of crosslinking to fibrin, to a greater extent than recombinant HSA. Conclusions Fusion protein XL5-HSA (DQMMLPWAVTLG4Y-HSAH6 was found to be more active as a substrate for FXIIIa-mediated transamidation than seven other candidate fusion proteins in

  1. Expression of Leukemia-Associated Nup98 Fusion Proteins Generates an Aberrant Nuclear Envelope Phenotype.

    Science.gov (United States)

    Fahrenkrog, Birthe; Martinelli, Valérie; Nilles, Nadine; Fruhmann, Gernot; Chatel, Guillaume; Juge, Sabine; Sauder, Ursula; Di Giacomo, Danika; Mecucci, Cristina; Schwaller, Jürg

    2016-01-01

    Chromosomal translocations involving the nucleoporin NUP98 have been described in several hematopoietic malignancies, in particular acute myeloid leukemia (AML). In the resulting chimeric proteins, Nup98's N-terminal region is fused to the C-terminal region of about 30 different partners, including homeodomain (HD) transcription factors. While transcriptional targets of distinct Nup98 chimeras related to immortalization are relatively well described, little is known about other potential cellular effects of these fusion proteins. By comparing the sub-nuclear localization of a large number of Nup98 fusions with HD and non-HD partners throughout the cell cycle we found that while all Nup98 chimeras were nuclear during interphase, only Nup98-HD fusion proteins exhibited a characteristic speckled appearance. During mitosis, only Nup98-HD fusions were concentrated on chromosomes. Despite the difference in localization, all tested Nup98 chimera provoked morphological alterations in the nuclear envelope (NE), in particular affecting the nuclear lamina and the lamina-associated polypeptide 2α (LAP2α). Importantly, such aberrations were not only observed in transiently transfected HeLa cells but also in mouse bone marrow cells immortalized by Nup98 fusions and in cells derived from leukemia patients harboring Nup98 fusions. Our findings unravel Nup98 fusion-associated NE alterations that may contribute to leukemogenesis.

  2. Overview of Fusion-Fission Hybrid Reactor Design Study in China

    International Nuclear Information System (INIS)

    Huang Jinhua; Feng Kaiming; Deng Baiquan; Deng, P.Zh.; Zhang Guoshu; Hu Gang; He Kaihui; Wu Yican; Qiu Lijian; Huang Qunying; Xiao Bingjia; Liu Xiaoping; Chen Yixue; Kong, M.H.

    2002-01-01

    The motivation for developing fusion-fission hybrid reactors is discussed in the context of electricity power requirements by 2050 in China. A detailed conceptual design of the Fusion Experimental Breeder (FEB) was developed from 1986-1995. The FEB has a subignited tokamak fusion core with a major radius of 4.0 m, a fusion power of 145 MW, and a fusion energy gain Q of 3. Based on this, an engineering outline design study of the FEB, FEB-E, has been performed. This design study is a transition from conceptual to engineering design in this research. The main results beyond that given in the detailed conceptual design are included in this paper, namely, the design studies of the blanket, divertor, test blanket, and tritium and environment issues. In-depth analyses have been performed to support the design. Studies of related advanced concepts such as the waste transmutation blanket concept and the spherical tokamak core concept are also presented

  3. Conceptual design of laser fusion reactor, SENRI-I - 1. concept and system design

    International Nuclear Information System (INIS)

    Ido, S.; Naki, S.; Norimatsu, T.

    1981-01-01

    Design features of a laser fusion reactor concept SENRI-I and new concepts are reviewed and discussed. The unique feature is the utilization of a magnetic field to guide and control the inner liquid Li flow. Basic requirements and typical parameters used in the design are presented. Items to be discussed are constitution of the system, performance of liquid Li flow, neutronics, thermo-electric cycle, fuel cycle and new concepts

  4. Design and evaluation of a laser fusion energy station for industrial applications

    International Nuclear Information System (INIS)

    Kok, K.D.; Bates, F.J.; Denning, R.S.; Triplett, M.B.; Waddell, J.D.

    1978-01-01

    The identification and development of long-term energy options is important in the continued growth of industry in the United States. Fusion and particularly laser fusion is one of the possible options. This paper applies the criteria used by industry in the selection of an energy source to the first of a series of conceptual designs for a laser fusion energy station. Several conclusions are presented including the constraints placed on the design by the criteria

  5. Trophoblast cell fusion and differentiation are mediated by both the protein kinase C and a pathways.

    Directory of Open Access Journals (Sweden)

    Waka Omata

    Full Text Available The syncytiotrophoblast of the human placenta is an epithelial barrier that interacts with maternal blood and is a key for the transfer of nutrients and other solutes to the developing fetus. The syncytiotrophoblast is a true syncytium and fusion of progenitor cytotrophoblasts is the cardinal event leading to the formation of this layer. BeWo cells are often used as a surrogate for cytotrophoblasts, since they can be induced to fuse, and then express certain differentiation markers associated with trophoblast syncytialization. Dysferlin, a syncytiotrophoblast membrane repair protein, is up-regulated in BeWo cells induced to fuse by treatment with forskolin; this fusion is thought to occur through cAMP/protein kinase A-dependent mechanisms. We hypothesized that dysferlin may also be up-regulated in response to fusion through other pathways. Here, we show that BeWo cells can also be induced to fuse by treatment with an activator of protein kinase C, and that this fusion is accompanied by increased expression of dysferlin. Moreover, a dramatic synergistic increase in dysferlin expression is observed when both the protein kinase A and protein kinase C pathways are activated in BeWo cells. This synergy in fusion is also accompanied by dramatic increases in mRNA for the placental fusion proteins syncytin 1, syncytin 2, as well as dysferlin. Dysferlin, however, was shown to be dispensable for stimulus-induced BeWo cell syncytialization, since dysferlin knockdown lines fused to the same extent as control cells. The classical trophoblast differentiation marker human chorionic gonadotropin was also monitored and changes in the expression closely parallel that of dysferlin in all of the experimental conditions employed. Thus different biochemical markers of trophoblast fusion behave in concert supporting the hypothesis that activation of both protein kinase C and A pathways lead to trophoblastic differentiation.

  6. Purification of CD47-streptavidin fusion protein from bacterial lysate using biotin-agarose affinity chromatography.

    Science.gov (United States)

    Salehi, Nasrin; Peng, Ching-An

    2016-07-08

    CD47 is a widely expressed transmembrane glycoprotein that modulates the activity of a plethora of immune cells via its extracellular domain. Therefore, CD47 plays important roles in the regulation of immune responses and may serve as targets for the development of immunotherapeutic agents. To make sure CD47 functionality is intact under the process of protein conjugation, CD47-streptavidin fusion protein was expressed and purified because it can easily bind to biotin-tagged materials via the unique biotin-streptavidin affinity. In this study, gene sequences of CD47 extracellular domain (CD47ECD) and core streptavidin (coreSA) with a total 834 bp were inserted into pET20b plasmid to construct recombinant plasmid encoding CD47-SA fusion gene. After bacteria transformation, the CD47-SA fusion protein was expressed by isopropyl-β-d-thiogalactopyranoside (IPTG) induction. The collected bacteria lysate was loaded on biotinylated agarose to proceed the purification of CD47-SA fusion protein. Due to the unexpected high affinity between biotin and coreSA, standard washing and elution approaches (e.g., varying pH, using biotin, and applying guanidine hydrochloride) reported for biotin-streptavidin affinity chromatography were not able to separate the target fusion protein. Instead, using low concentration of the non-ionic detergent Triton X-100 followed with alkaline buffer could efficiently weaken the binding between biotin and coreSA, thereby eluting out CD47-SA fusion protein from the biotin agarose column. The purified CD47-SA fusion protein was further characterized by molecular biology methods and its antiphagocytic functionality was confirmed by the phagocytosis assay. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:949-958, 2016. © 2016 American Institute of Chemical Engineers.

  7. Nuclear challenges and progress in designing stellarator fusion power plants

    International Nuclear Information System (INIS)

    El-Guebaly, L.A.; Wilson, P.; Henderson, D.; Sawan, M.; Sviatoslavsky, G.; Tautges, T.; Slaybaugh, R.; Kiedrowski, B.; Ibrahim, A.

    2008-01-01

    Over the past 5-6 decades, stellarator power plants have been studied in the US, Europe, and Japan as an alternate to the mainline magnetic fusion tokamaks, offering steady-state operation and eliminating the risk of plasma disruptions. The earlier 1980s studies suggested large-scale stellarator power plants with an average major radius exceeding 20 m. The most recent development of the compact stellarator concept delivered ARIES-CS - a compact stellarator with 7.75 m average major radius, approaching that of tokamaks. For stellarators, the most important engineering parameter that determines the machine size and cost is the minimum distance between the plasma boundary and mid-coil. Accommodating the breeding blanket and necessary shield within this distance to protect the ARIES-CS superconducting magnet represents a challenging task. Selecting the ARIES-CS nuclear and engineering parameters to produce an economic optimum, modeling the complex geometry for 3D nuclear analysis to confirm the key parameters, and minimizing the radwaste stream received considerable attention during the design process. These engineering design elements combined with advanced physics helped enable the compact stellarator to be a viable concept. This paper provides a brief historical overview of the progress in designing stellarator power plants and a perspective on the successful integration of the nuclear activity into the final ARIES-CS configuration

  8. Remote maintenance design for Fusion Experimental Reactor (FER)

    International Nuclear Information System (INIS)

    Tachikawa, K.; Iida, H.; Nishio, S.; Tone, T.; Aota, T.; Iwamoto, T.; Niikura, S.; Nishizawa, H.

    1984-01-01

    Design of Fusion Experimental Reactor, FER, has been conducted by Japan Atomic Energy Research Institute (JAERI) since 1981. Two typical reactors can be classified in general from the viewpoints of remote maintenance among four design concepts of FER. In the case of the type 1 FER, the torus module consists of shield structure and blanket, and the connective joints between toruses provided at the outer region of the reactor. As for the type 2 FER, the shield structure is joined with the vacuum cryostat, and only the blanket module is allowed to move, but connection between toruses are located in the inner region of the reactor. Comparing type 1 with type 2 FER, this paper describes on the remote maintenance of FER including reactor configurations, work procedures, remote systems/equipments, repairing facility and future R and D problems. Reviewing design studies and investigation for the existing robotics technologies, R and D for FER remote maintenance technology should be performed under the reasonable long-term program. The main items of remote technology required to start urgently are multi-purpose manipulator system with performance of dextrousity, tele-viewing system which reduces operator fatigue and remote tests for commercially available components

  9. Applying design principles to fusion reactor configurations for propulsion in space

    International Nuclear Information System (INIS)

    Carpenter, S.A.; Deveny, M.E.; Schulze, N.R.

    1993-01-01

    The application of fusion power to space propulsion requires rethinking the engineering-design solution to controlled-fusion energy. Whereas the unit cost of electricity (COE) drives the engineering-design solution for utility-based fusion reactor configurations; initial mass to low earth orbit (IMLEO), specific jet power (kW(thrust)/kg(engine)), and reusability drive the engineering-design solution for successful application of fusion power to space propulsion. Three design principles (DP's) were applied to adapt and optimize three candidate-terrestrial-fusion-reactor configurations for propulsion in space. The three design principles are: provide maximum direct access to space for waste radiation, operate components as passive radiators to minimize cooling-system mass, and optimize the plasma fuel, fuel mix, and temperature for best specific jet power. The three candidate terrestrial fusion reactor configurations are: the thermal barrier tandem mirror (TBTM), field reversed mirror (FRM), and levitated dipole field (LDF). The resulting three candidate space fusion propulsion systems have their IMLEO minimized and their specific jet power and reusability maximized. A preliminary rating of these configurations was performed, and it was concluded that the leading engineering-design solution to space fusion propulsion is a modified TBTM that we call the Mirror Fusion Propulsion System (MFPS)

  10. Chemical Hazards and Safety Issues in Fusion Safety Design

    International Nuclear Information System (INIS)

    Cadwallader, L.C.

    2003-01-01

    Radiological inventory releases have dominated accident consequences for fusion; these consequences are important to analyze and are generally the most severe result of a fusion facility accident event. However, the advent of, or plan for, large-scale usage of some toxic materials poses the additional hazard of chemical exposure from an accident event. Examples of toxic chemicals are beryllium for magnetic fusion and fluorine for laser fusion. Therefore, chemical exposure consequences must also be addressed in fusion safety assessment. This paper provides guidance for fusion safety analysis. US Department of Energy (DOE) chemical safety assessment practices for workers and the public are reviewed. The US Environmental Protection Agency (EPA) has published some guidance on public exposure to releases of mixtures of chemicals, this guidance has been used to create an initial guideline for treating mixed radiological and toxicological releases in fusion; for example, tritiated hazardous dust from a tokamak vacuum vessel. There is no convenient means to judge the hazard severity of exposure to mixed materials. The chemical fate of mixed material constituents must be reviewed to determine if there is a separate or combined radiological and toxicological carcinogenesis, or if other health threats exist with radiological carcinogenesis. Recommendations are made for fusion facility chemical safety evaluation and safety guidance for protecting the public from chemical releases, since such levels are not specifically identified in the DOE fusion safety standard

  11. Radiological safety design considerations for fusion research experiments

    International Nuclear Information System (INIS)

    Crase, K.W.; Singh, M.S.

    1979-01-01

    A wide variety of fusion research experiments are in the planning or construction stages. Two such experiments, the Nova Laser Fusion Facility and the Mirror Fusion Test Facility (MFTF), are currently under construction at Lawrence Livermore Laboratory. Although the plasma chamber vault for MFTF and the Nova target room will have thick concrete walls and roofs, the radiation safety problems are made complex by the numerous requirements for shield wall penetrations. This paper addresses radiation safety considerations for the MFTF and Nova experiments, and the need for integrated safety considerations and safety technology development during the planning stages of fusion experiments

  12. Macrophage fusion is controlled by the cytoplasmic protein tyrosine phosphatase PTP-PEST/PTPN12.

    Science.gov (United States)

    Rhee, Inmoo; Davidson, Dominique; Souza, Cleiton Martins; Vacher, Jean; Veillette, André

    2013-06-01

    Macrophages can undergo cell-cell fusion, leading to the formation of multinucleated giant cells and osteoclasts. This process is believed to promote the proteolytic activity of macrophages toward pathogens, foreign bodies, and extracellular matrices. Here, we examined the role of PTP-PEST (PTPN12), a cytoplasmic protein tyrosine phosphatase, in macrophage fusion. Using a macrophage-targeted PTP-PEST-deficient mouse, we determined that PTP-PEST was not needed for macrophage differentiation or cytokine production. However, it was necessary for interleukin-4-induced macrophage fusion into multinucleated giant cells in vitro. It was also needed for macrophage fusion following implantation of a foreign body in vivo. Moreover, in the RAW264.7 macrophage cell line, PTP-PEST was required for receptor activator of nuclear factor kappa-B ligand (RANKL)-triggered macrophage fusion into osteoclasts. PTP-PEST had no impact on expression of fusion mediators such as β-integrins, E-cadherin, and CD47, which enable macrophages to become fusion competent. However, it was needed for polarization of macrophages, migration induced by the chemokine CC chemokine ligand 2 (CCL2), and integrin-induced spreading, three key events in the fusion process. PTP-PEST deficiency resulted in specific hyperphosphorylation of the protein tyrosine kinase Pyk2 and the adaptor paxillin. Moreover, a fusion defect was induced upon treatment of normal macrophages with a Pyk2 inhibitor. Together, these data argue that macrophage fusion is critically dependent on PTP-PEST. This function is seemingly due to the ability of PTP-PEST to control phosphorylation of Pyk2 and paxillin, thereby regulating cell polarization, migration, and spreading.

  13. Conceptual design studies of experimental and demonstration fusion reactors

    International Nuclear Information System (INIS)

    1978-01-01

    Since 1973 the FINTOR Group has been involved in conceptual design studies of TOKAMAK-type fusion reactors to precede the construction of a prototype power reactor plant. FINTOR-1 was the first conceptual design aimed at investigating the main physics and engineering constraints on a minimum-size (both dimensions and thermal power) tokamak experimental reactor. The required plasma energy confinement time as evaluated by various power balance models was compared with the values resulting from different transport models. For the reference design, an energy confinement time ten times smaller than neoclassical was assumed. This also implied a rather high (thermally stable) working temperature (above 20 keV) for the reactor. Other relevant points of the design were: circular plasma cross section, single-null axisymmetric divertor; lithium breeder, stainless steel structures, helium coolant; modular blanket and shield structure; copper-stabilized, superconducting Nb-Ti toroidal field and divertor coils; vertical field and transformer coils inside the toroidal coils; vacuum-tight containment vessel. Solutions involving air and iron transformer cores were compared. These assumptions led to a minimum size reactor with a thermal power of about 100MW and rather large dimensions (major radius of about 9m) similar to those of full-scale power reactors considered in other conceptual studies. The FINTOR-1 analysis was completed by the end of 1976. In 1977 a conceptual design of a Demonstration Power Reactor Plant (FINTOR-D) was started. In this study the main working assumptions differing from those of FINTOR-1 are: non-circular plasma cross section; plasma confinement compatible with trapped ion instabilities; cold (gas) blanket sufficient for wall protection (no divertor); wall loading between 1-3MW/m 2 and thermal power of a few GW. (author)

  14. Structure of the cleavage-activated prefusion form of the parainfluenza virus 5 fusion protein.

    Science.gov (United States)

    Welch, Brett D; Liu, Yuanyuan; Kors, Christopher A; Leser, George P; Jardetzky, Theodore S; Lamb, Robert A

    2012-10-09

    The paramyxovirus parainfluenza virus 5 (PIV5) enters cells by fusion of the viral envelope with the plasma membrane through the concerted action of the fusion (F) protein and the receptor binding protein hemagglutinin-neuraminidase. The F protein folds initially to form a trimeric metastable prefusion form that is triggered to undergo large-scale irreversible conformational changes to form the trimeric postfusion conformation. It is thought that F refolding couples the energy released with membrane fusion. The F protein is synthesized as a precursor (F0) that must be cleaved by a host protease to form a biologically active molecule, F1,F2. Cleavage of F protein is a prerequisite for fusion and virus infectivity. Cleavage creates a new N terminus on F1 that contains a hydrophobic region, known as the FP, which intercalates target membranes during F protein refolding. The crystal structure of the soluble ectodomain of the uncleaved form of PIV5 F is known; here we report the crystal structure of the cleavage-activated prefusion form of PIV5 F. The structure shows minimal movement of the residues adjacent to the protease cleavage site. Most of the hydrophobic FP residues are buried in the uncleaved F protein, and only F103 at the newly created N terminus becomes more solvent-accessible after cleavage. The conformational freedom of the charged arginine residues that compose the protease recognition site increases on cleavage of F protein.

  15. Protein-Protein Docking in Drug Design and Discovery.

    Science.gov (United States)

    Kaczor, Agnieszka A; Bartuzi, Damian; Stępniewski, Tomasz Maciej; Matosiuk, Dariusz; Selent, Jana

    2018-01-01

    Protein-protein interactions (PPIs) are responsible for a number of key physiological processes in the living cells and underlie the pathomechanism of many diseases. Nowadays, along with the concept of so-called "hot spots" in protein-protein interactions, which are well-defined interface regions responsible for most of the binding energy, these interfaces can be targeted with modulators. In order to apply structure-based design techniques to design PPIs modulators, a three-dimensional structure of protein complex has to be available. In this context in silico approaches, in particular protein-protein docking, are a valuable complement to experimental methods for elucidating 3D structure of protein complexes. Protein-protein docking is easy to use and does not require significant computer resources and time (in contrast to molecular dynamics) and it results in 3D structure of a protein complex (in contrast to sequence-based methods of predicting binding interfaces). However, protein-protein docking cannot address all the aspects of protein dynamics, in particular the global conformational changes during protein complex formation. In spite of this fact, protein-protein docking is widely used to model complexes of water-soluble proteins and less commonly to predict structures of transmembrane protein assemblies, including dimers and oligomers of G protein-coupled receptors (GPCRs). In this chapter we review the principles of protein-protein docking, available algorithms and software and discuss the recent examples, benefits, and drawbacks of protein-protein docking application to water-soluble proteins, membrane anchoring and transmembrane proteins, including GPCRs.

  16. Design windows of laser fusion power plants and conceptual design of laser-diode pumped slab laser

    International Nuclear Information System (INIS)

    Kozaki, Y.; Eguchi, T.; Izawa, Y.

    1999-01-01

    An analysis of the design space available to laser fusion power plants has been carried out, in terms of design key parameters such as target gain, laser energy and laser repetition rate, the number of fusion react ion chambers, and plant size. The design windows of economically attractive laser fusion plants is identified with the constraints of key design parameters and the cost conditions. Especially, for achieving high repetition rate lasers, we have proposed and designed a diode-pumped solid-state laser driver which consists of water-cooled zig-zag path slab amplifiers. (author)

  17. Cooling System Design Options for a Fusion Reactor

    Science.gov (United States)

    Natalizio, Antonio; Collén, Jan; Vieider, Gottfried

    1997-06-01

    The objective of a fusion power reactor is to produce electricity safely and reliably. Accordingly, the design, objective of the heat transport system is to optimize power production, safety, and reliability. Such an optimization process, however, is constrained by many factors, including, among others: public safety, worker safety, steam cycle efficiency, reliability, and cost. As these factors impose conflicting requirements, there is a need to find an optimum design solution, i.e., one that satisfies all requirements, but not necessarily each requirement optimally. The SEAFP reactor study developed helium-cooled and water-cooled models for assessment purposes. Among other things, the current study demonstrates that neither model offers an optimum solution. Helium cooling offers a high steam cycle efficiency but poor reliability for the cooling of high heat flux components (divertor and first wall). Alternatively, water cooling offers a low steam cycle efficiency, but reasonable reliability for the cooling of such components. It is concluded that an optimum solution includes helium cooling of low heat flux components and water cooling of high heat flux components. Relative to the SEAFP helium model, this hybrid system enhances safety and reliability, while retaining the high steam cycle efficiency of that model.

  18. Conceptual design study of fusion experimental reactor (FY86 FER)

    International Nuclear Information System (INIS)

    Nakashima, Kunihiko; Okano, Kunihiko; Miyamoto, Kazuhiro.

    1987-09-01

    This report describes the results of a conceptual study on the RF system in the typical candidates for the Fusion Experimental Reactor (FER), which were picked out through the '86FER scoping studies. According to the FER operation scenario, three RF systems, that is, ICRF (heating), LHRF (current drive and heating), ECRF (auxiliary heating) were studied. Main concern in these RF systems is the launcher, which may be so designed that required power match the geometrical constraints of the reactor. Then studies were concentrated on the launcher configuration. A prug-in concept of the launcher was adopted in each system and vacancies except transmission space were filled with water. The ICRF launcher had the 2 x 2 loop arrays antenna and the faraday shield area of 1.5 m x 1 m to provide a power of 20 MW. The LHRF launcher had the grillantenna with 28 x 8 open waveguides, and included multi junction-type power splitters which were connected to 56 transmission wave guides. The grild was designed to have two functions of current drive and heating, and provide a power of 20 MW each. The ECRF launcher had a boundle of open wave guides which a reflection mirror each, and three plain mirrors. Assuming a oscillator unit size of 200 kW, it had 40 oversized wave guides to provide a power of 3 MW. (author)

  19. Development of design system of manipulator for fusion reactor maintenance

    International Nuclear Information System (INIS)

    Ida, Toshio; Niikura, Setsuo; Ishiguro, Akiko; Yamada, Masao; Matsuoka, Fushiki

    1989-01-01

    A program which supports designers of a manipulator for in-vessel maintenance of a fusion reactor has been developed. The main purpose of this program is to provide the designer with a promising manipulator specification by furnishing useful information. It combines a technique of knowledge engineering with numerical solutions. This program consists of three parts: The first part is to generate candidates for the manipulator using a knowledge base; the second is to evaluate both static and dynamic properties of each candidate through numerical simulation of the maintenance task; and the third is to select the candidates having better performance and feasibility on the basis of the simulation data and knowledge base. The feasibility of the method used in this program is confirmed by the preliminary application. This application also emphasizes the importance of the knowledge base for the candidate generation and selection. Although the degree of freedom of the manipulator is restricted to less than 7 degrees in this study, further development of the capability of manipulator simulation (seven or more degrees of freedom) will enhance the effectiveness of this program. (orig.)

  20. Conceptual design of the fusion-driven subcritical system FDS-I

    International Nuclear Information System (INIS)

    Wu, Y.; Zheng, S.; Zhu, X.; Wang, W.; Wang, H.; Liu, S.; Bai, Y.; Chen, H.; Hu, L.; Chen, M.; Huang, Q.; Huang, D.; Zhang, S.; Li, J.; Chu, D.; Jiang, J.; Song, Y.

    2006-01-01

    The fusion-driven subcritical system (named FDS-I) was previously proposed as an intermediate step toward the final application of fusion energy. A conceptual design of the FDS-I is presented, which consists of the fusion neutron driver with relatively easy-achieved plasma parameters, and the He-gas/liquid lithium-lead Dual-cooled subcritical Waste Transmutation (DWT) blanket used to transmute long-lived radioactive wastes and to generate energy on the basis of self-sustainable fission and fusion fuel cycle. An overview of the FDS-I is given and the specifications of the design analysis are summarized

  1. Updated reference design of a liquid metal cooled tandem mirror fusion breeder

    Energy Technology Data Exchange (ETDEWEB)

    Berwald, D.H.; Whitley, R.H.; Garner, J.K.; Gromada, R.J.; McCarville, T.J.; Moir, R.W.; Lee, J.D.; Bandini, B.R.; Fulton, F.J.; Wong, C.P.C.; Maya, I.; Hoot, C.G.; Schultz, K.R.; Miller, L.G.; Beeston, J.M.; Harris, B.L.; Westman, R.A.; Ghoniem, N.M.; Orient, G.; Wolfer, M.; DeVan, J.H.; Torterelli, P.

    1985-09-01

    Detailed studies of key techinical issues for liquid metal cooled fusion breeder (fusion-fission hybrid blankets) have been performed during the period 1983-4. Based upon the results of these studies, the 1982 reference liquid metal cooled tandem mirror fusion breeder blanket design was updated and is described. The updated reference blankets provides increased breeding and lower technological risk in comparison with the original reference blanket. In addition to the blanket design revisions, a plant concept, cost, and fuel cycle economics assessment is provided. The fusion breeder continues to promise an economical source of fissile fuel for the indefinite future.

  2. Updated reference design of a liquid metal cooled tandem mirror fusion breeder

    International Nuclear Information System (INIS)

    Berwald, D.H.; Whitley, R.H.; Garner, J.K.

    1985-09-01

    Detailed studies of key techinical issues for liquid metal cooled fusion breeder (fusion-fission hybrid blankets) have been performed during the period 1983-4. Based upon the results of these studies, the 1982 reference liquid metal cooled tandem mirror fusion breeder blanket design was updated and is described. The updated reference blankets provides increased breeding and lower technological risk in comparison with the original reference blanket. In addition to the blanket design revisions, a plant concept, cost, and fuel cycle economics assessment is provided. The fusion breeder continues to promise an economical source of fissile fuel for the indefinite future

  3. Antibody-cytokine fusion proteins for improving efficacy and safety of cancer therapy.

    Science.gov (United States)

    Valedkarimi, Zahra; Nasiri, Hadi; Aghebati-Maleki, Leili; Majidi, Jafar

    2017-11-01

    Cytokines are key players in the regulation of immune responses both in physiological and pathological states. A number of cytokines have been evaluated in clinical trials and shown promising results in the treatment of different malignancies. Despite this, the clinical application of these molecules may be plagued by undesirable side effects The development of recombinant antibody-cytokine fusion proteins, which offer a means for target delivery of cytokines toward the tumor site, has significantly improved the therapeutic index of these immunomodulatory molecules. Selective tumor localization is provided by the monoclonal antibody component of the fusion protein that binds to the molecules present on the surface of tumor cells or accumulated preferentially in the diseased site. In this manner, the cytokine element is specifically located at the tumor site and can stimulate immune cells with appropriate cytokine receptors. Over the recent years, several antibody-cytokine fusion proteins have been developed with the capacity to target a wide variety of cancers whose application, in some cases, has led to complete rejection of the tumor. These findings support the notion that antibody-cytokine fusion proteins represent huge potential for cancer therapy. This review presents an overview of the advances made in the field of targeted cytokine delivery, which is made possible by genetically engineering antibody-cytokine fusion proteins. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

  4. IGF1 is a common target gene of Ewing's sarcoma fusion proteins in mesenchymal progenitor cells.

    Directory of Open Access Journals (Sweden)

    Luisa Cironi

    Full Text Available BACKGROUND: The EWS-FLI-1 fusion protein is associated with 85-90% of Ewing's sarcoma family tumors (ESFT, the remaining 10-15% of cases expressing chimeric genes encoding EWS or FUS fused to one of several ets transcription factor family members, including ERG-1, FEV, ETV1 and ETV6. ESFT are dependent on insulin-like growth factor-1 (IGF-1 for growth and survival and recent evidence suggests that mesenchymal progenitor/stem cells constitute a candidate ESFT origin. METHODOLOGY/PRINCIPAL FINDINGS: To address the functional relatedness between ESFT-associated fusion proteins, we compared mouse progenitor cell (MPC permissiveness for EWS-FLI-1, EWS-ERG and FUS-ERG expression and assessed the corresponding expression profile changes. Whereas all MPC isolates tested could stably express EWS-FLI-1, only some sustained stable EWS-ERG expression and none could express FUS-ERG for more than 3-5 days. Only 14% and 4% of the total number of genes that were respectively induced and repressed in MPCs by the three fusion proteins were shared. However, all three fusion proteins, but neither FLI-1 nor ERG-1 alone, activated the IGF1 promoter and induced IGF1 expression. CONCLUSION/SIGNIFICANCE: Whereas expression of different ESFT-associated fusion proteins may require distinct cellular microenvironments and induce transcriptome changes of limited similarity, IGF1 induction may provide one common mechanism for their implication in ESFT pathogenesis.

  5. Egg CD9 protein tides correlated with sperm oscillations tune the gamete fusion ability in mammal.

    Science.gov (United States)

    Ravaux, Benjamin; Favier, Sophie; Perez, Eric; Gourier, Christine

    2018-01-23

    Mammalian fertilization involves membrane events -adhesion, fusion, sperm engulfment, membrane block to polyspermy- whose causes remain largely unknown. Recently, specific oscillations of the sperm in contact with the egg were shown to be necessary for fusion. Using a microfluidic chip to impose the venue for the encounter of two gametes allowed real-time observation of the membrane remodelling occurring at the sperm/egg interface. The spatiotemporal mapping of egg CD9 revealed that this protein concentrates at the egg/sperm interface as a result of sperm oscillations, until a CD9-rich platform is nucleated on which fusion immediately takes place. Within 2 to 5 minutes after fusion, most of the CD9 leaves the egg for the external aqueous medium. Then an egg membrane wave engulfs the sperm head in approximately 25 minutes. These results show that sperm oscillations initiate the CD9 recruitment that causes gamete fusion after which CD9 and associated proteins leave the membrane in a process likely to contribute to block polyspermy. They highlight that the gamete fusion story in mammals is an unexpected interplay between mechanical constraints and proteins. © The Author(s) (2018). Published by Oxford University Press on behalf of Journal of Molecular Cell Biology, IBCB, SIBS, CAS. All rights reserved.

  6. Biodistribution and tumor imaging of an anti-CEA single-chain antibody-albumin fusion protein

    International Nuclear Information System (INIS)

    Yazaki, Paul J.; Kassa, Thewodros; Cheung, Chia-wei; Crow, Desiree M.; Sherman, Mark A.; Bading, James R.; Anderson, Anne-Line J.; Colcher, David; Raubitschek, Andrew

    2008-01-01

    Albumin fusion proteins have demonstrated the ability to prolong the in vivo half-life of small therapeutic proteins/peptides in the circulation and thereby potentially increase their therapeutic efficacy. To evaluate if this format can be employed for antibody-based imaging, an anticarcinoembryonic antigen (CEA) single-chain antibody(scFv)-albumin fusion protein was designed, expressed and radiolabeled for biodistribution and imaging studies in athymic mice bearing human colorectal carcinoma LS-174T xenografts. The [ 125 I]-T84.66 fusion protein demonstrated rapid tumor uptake of 12.3% injected dose per gram (ID/g) at 4 h that reached a plateau of 22.7% ID/g by 18 h. This was a dramatic increase in tumor uptake compared to 4.9% ID/g for the scFv alone. The radiometal [ 111 In]-labeled version resulted in higher tumor uptake, 37.2% ID/g at 18 h, which persisted at the tumor site with tumor: blood ratios reaching 18:1 and with normal tissues showing limited uptake. Based on these favorable imaging properties, a pilot [ 64 Cu]-positron emission tomography imaging study was performed with promising results. The anti-CEA T84.66 scFv-albumin fusion protein demonstrates highly specific tumor uptake that is comparable to cognate recombinant antibody fragments. The radiometal-labeled version, which shows lower normal tissue accumulation than these recombinant antibodies, provides a promising and novel platform for antibody-based imaging agents

  7. Induction of protein body formation in plant leaves by elastin-like polypeptide fusions

    Directory of Open Access Journals (Sweden)

    Joensuu Jussi J

    2009-08-01

    Full Text Available Abstract Background Elastin-like polypeptides are synthetic biopolymers composed of a repeating pentapeptide 'VPGXG' sequence that are valuable for the simple non-chromatographic purification of recombinant proteins. In addition, elastin-like polypeptide fusions have been shown to enhance the accumulation of a range of different recombinant proteins in plants, thus addressing the major limitation of plant-based expression systems, which is a low production yield. This study's main objectives were to determine the general utility of elastin-like polypeptide protein fusions in various intracellular compartments and to elucidate elastin-like polypeptide's mechanism of action for increasing recombinant protein accumulation in the endoplasmic reticulum of plants. Results The effect of elastin-like polypeptide fusions on the accumulation of green fluorescent protein targeted to the cytoplasm, chloroplasts, apoplast, and endoplasmic reticulum was evaluated. The endoplasmic reticulum was the only intracellular compartment in which an elastin-like polypeptide tag was shown to significantly enhance recombinant protein accumulation. Interestingly, endoplasmic reticulum-targeted elastin-like polypeptide fusions induced the formation of a novel type of protein body, which may be responsible for elastin-like polypeptide's positive effect on recombinant protein accumulation by excluding the heterologous protein from normal physiological turnover. Although expressed in the leaves of plants, these novel protein bodies appeared similar in size and morphology to the prolamin-based protein bodies naturally found in plant seeds. The elastin-like polypeptide-induced protein bodies were highly mobile organelles, exhibiting various dynamic patterns of movement throughout the cells, which were dependent on intact actin microfilaments and a functional actomyosin motility system. Conclusion An endoplasmic reticulum-targeted elastin-like polypeptide fusion approach

  8. Characterization of the fusion core in zebrafish endogenous retroviral envelope protein

    Energy Technology Data Exchange (ETDEWEB)

    Shi, Jian [State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan, Hubei 430072 (China); State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, Hubei 430071 (China); Zhang, Huaidong [CAS Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, Hubei 430071 (China); Gong, Rui, E-mail: gongr@wh.iov.cn [CAS Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, Hubei 430071 (China); Xiao, Gengfu, E-mail: xiaogf@wh.iov.cn [State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan, Hubei 430072 (China); State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, Hubei 430071 (China)

    2015-05-08

    Zebrafish endogenous retrovirus (ZFERV) is the unique endogenous retrovirus in zebrafish, as yet, containing intact open reading frames of its envelope protein gene in zebrafish genome. Similarly, several envelope proteins of endogenous retroviruses in human and other mammalian animal genomes (such as syncytin-1 and 2 in human, syncytin-A and B in mouse) were identified and shown to be functional in induction of cell–cell fusion involved in placental development. ZFERV envelope protein (Env) gene appears to be also functional in vivo because it is expressible. After sequence alignment, we found ZFERV Env shares similar structural profiles with syncytin and other type I viral envelopes, especially in the regions of N- and C-terminal heptad repeats (NHR and CHR) which were crucial for membrane fusion. We expressed the regions of N + C protein in the ZFERV Env (residues 459–567, including predicted NHR and CHR) to characterize the fusion core structure. We found N + C protein could form a stable coiled-coil trimer that consists of three helical NHR regions forming a central trimeric core, and three helical CHR regions packing into the grooves on the surface of the central core. The structural characterization of the fusion core revealed the possible mechanism of fusion mediated by ZFERV Env. These results gave comprehensive explanation of how the ancient virus infects the zebrafish and integrates into the genome million years ago, and showed a rational clue for discovery of physiological significance (e.g., medicate cell–cell fusion). - Highlights: • ZFERV Env shares similar structural profiles with syncytin and other type I viral envelopes. • The fusion core of ZFERV Env forms stable coiled-coil trimer including three NHRs and three CHRs. • The structural mechanism of viral entry mediated by ZFERV Env is disclosed. • The results are helpful for further discovery of physiological function of ZFERV Env in zebrafish.

  9. Conceptual design of a mirror reactor for a fusion engineering research facility (FERF)

    International Nuclear Information System (INIS)

    Batzer, T.H.; Burleigh, R.C.; Carlson, G.A.; Dexter, W.L.; Hamilton, G.W.; Harvey, A.R.; Hickman, R.G.; Hoffman, M.A.; Hooper, E.B. Jr.; Moir, R.W.; Nelson, R.L.; Pittenger, L.C.; Smith, B.H.; Taylor, C.E.; Werner, R.W.; Wilcox, T.P.

    1975-01-01

    A conceptual design is presented for a small mirror fusion reactor for a Fusion Engineering Research Facility (FERF). The reactor produces 3.4 MW of fusion power and a useful neutron flux of about 10 14 n.cm -2 .s -1 . Superconducting ''yin-yang'' coils are used, and the plasma is sustained by injection of energetic neutral D 0 and T 0 . Conceptual layouts are given for the reactor, its major components, and supporting facilities. (author)

  10. Review of fission-fusion pellet designs and inertial confinement system studies at EIR

    Energy Technology Data Exchange (ETDEWEB)

    Seifriz, W [Eidgenoessisches Inst. fuer Reaktorforschung, Wuerenlingen (Switzerland)

    1978-01-01

    The article summarizes the work done so far at the Swiss Federal Institute for Reactor Research (EIR) in the field of the inertial confinement fusion technique. The following subjects are reviewed: a) fission fusion pellet designs using fissionable triggers, b) uranium tampered pellets, c) tampered pellets recycling unwanted actinide wastes from fission reactors in beam-driven micro-explosion reactors, and d) symbiotic fusion/fission reactor studies.

  11. Cleaved thioredoxin fusion protein enables the crystallization of poorly soluble ERα in complex with synthetic ligands

    International Nuclear Information System (INIS)

    Cura, Vincent; Gangloff, Monique; Eiler, Sylvia; Moras, Dino; Ruff, Marc

    2007-01-01

    A new crystallization strategy: the presence of cleaved thioredoxin fusion is critical for crystallization of the estrogen nuclear receptor ligand binding domain in complex with synthetic ligands. This novel technique should be regarded as an interesting alternative for crystallization of difficult proteins. The ligand-binding domain (LBD) of human oestrogen receptor α was produced in Escherichia coli as a cleavable thioredoxin (Trx) fusion in order to improve solubility. Crystallization trials with either cleaved and purified LBD or with the purified fusion protein both failed to produce crystals. In another attempt, Trx was not removed from the LBD after endoproteolytic cleavage and its presence promoted nucleation and subsequent crystal growth, which allowed the structure determination of two different LBD–ligand–coactivator peptide complexes at 2.3 Å resolution. This technique is likely to be applicable to other low-solubility proteins

  12. Mechanistic insight provided by glutaredoxin within a fusion to redox-sensitive yellow fluorescent protein

    DEFF Research Database (Denmark)

    Björnberg, Olof; Østergaard, Henrik; Winther, Jakob R

    2006-01-01

    Redox-sensitive yellow fluorescent protein (rxYFP) contains a dithiol disulfide pair that is thermodynamically suitable for monitoring intracellular glutathione redox potential. Glutaredoxin 1 (Grx1p) from yeast is known to catalyze the redox equilibrium between rxYFP and glutathione, and here, we...... have generated a fusion of the two proteins, rxYFP-Grx1p. In comparison to isolated subunits, intramolecular transfer of reducing equivalents made the fusion protein kinetically superior in reactions with glutathione. The rate of GSSG oxidation was thus improved by a factor of 3300. The reaction...... separately and in the fusion. This could not be ascribed to the lack of an unproductive side reaction to glutaredoxin disulfide. Instead, slower alkylation kinetics with iodoacetamide indicates a better leaving-group capability of the remaining cysteine residue, which can explain the increased activity....

  13. High-level expression of soluble recombinant proteins in Escherichia coli using an HE-maltotriose-binding protein fusion tag.

    Science.gov (United States)

    Han, Yingqian; Guo, Wanying; Su, Bingqian; Guo, Yujie; Wang, Jiang; Chu, Beibei; Yang, Guoyu

    2018-02-01

    Recombinant proteins are commonly expressed in prokaryotic expression systems for large-scale production. The use of genetically engineered affinity and solubility enhancing fusion proteins has increased greatly in recent years, and there now exists a considerable repertoire of these that can be used to enhance the expression, stability, solubility, folding, and purification of their fusion partner. Here, a modified histidine tag (HE) used as an affinity tag was employed together with a truncated maltotriose-binding protein (MBP; consisting of residues 59-433) from Pyrococcus furiosus as a solubility enhancing tag accompanying a tobacco etch virus protease-recognition site for protein expression and purification in Escherichia coli. Various proteins tagged at the N-terminus with HE-MBP(Pyr) were expressed in E. coli BL21(DE3) cells to determine expression and solubility relative to those tagged with His6-MBP or His6-MBP(Pyr). Furthermore, four HE-MBP(Pyr)-fused proteins were purified by immobilized metal affinity chromatography to assess the affinity of HE with immobilized Ni 2+ . Our results showed that HE-MBP(Pyr) represents an attractive fusion protein allowing high levels of soluble expression and purification of recombinant protein in E. coli. Copyright © 2017 Elsevier Inc. All rights reserved.

  14. Radiological safety design considerations for a laser-fusion facility

    International Nuclear Information System (INIS)

    Singh, M.S.

    1977-01-01

    Detailed neutronics and photonics calculations have been performed for analyzing prompt and residual radiations and required shielding associated with the design of a laser-fusion facility with a nominal yield of 10 19 neutrons per D--T burn pulse. The standard Livermore Monte Carlo codes and nuclear data cross section libraries were used in calculations. The Bateman equation was used to calculate the accumulation and decay of radionuclide chain products. A number of activation sensitivity experiments were conducted and the results were found to be in very good agreement within 10 percent of those calculated. It has been found that neutron yields of 2 x 10 19 per day can be conducted continuously if the reactor chamber is Kevlar-epoxy or silica, the primary shield is 0.60-m of water immediately on the chamber, and the building concrete is 1.80 m thick. These precautions result in dose equivalents below the primary protection limits inside the target room after a few hours of cool-down per each 10 19 pulse, 10 percent of the primary protection limits immediately outside the target room, and 1 percent of the natural background level at the nearest site boundary

  15. Conceptual design study of fusion experimental reactor (FY86FER)

    International Nuclear Information System (INIS)

    Nakashima, Kunihiko; Yamamoto, Shin; Ohara, Yoshihiro; Watanabe, Kazuhiro; Mizuno, Makoto; Araki, Masanori; Uede, Taisei; Okano, Kunihiko.

    1987-09-01

    This report describes the results of applicability studies for the negative ion-based neutral beam injector to the Fusion Experimental Reactor (FER). The operation scenario of FER has been proposed to adopt the neutral injection method as one of candidates, which has three functions of heating, current drive and profile control. One of the fundamental requirements is the tangential injection of the neutral beam. For neutral beam injectors, three port sections are available. Supposing to adopt the beam line with the straight long neutralizer which has been designed in JAERI, the geometrical arrangement was determined so as to avoid any trouble to the reactor structure. The conceptual study for major components which compose the beam line system was carried out including the estimation of the neutron streaming. The power supply system was studied also and the work was concentrated on the acceleration power supply which requires the output voltage of 500 kV and fast cut-off time. A basic concept, in which a inverter with a AC switch is used and the frequency of the supplied AC line is increased was proposed. In these works, the configuration of the neutral beam injection system was detailed and it was shown that the beam line seems to be well implemented with the geometrical constraints related to the reactor configuration. (author)

  16. The Modular Point Design for Heavy Ion Fusion

    International Nuclear Information System (INIS)

    Yu, S.S.; Barnard, J.J.; Briggs, R.J.; Callahan, D.; Celata, C.M.; Chao, L.; Davidson, R.; Debonnel, C.S.; Eylon, S.; Friedman, A.; Henestroza, E.; Kaganovich, I.; Kwan, J.W.; Lee, E.P.; Leitner, M.; Logan, B.G.; Meier, W.; Peterson, P.F.; Reginato, L.; Rose, D.; Roy, P.; Waldron, W.; Welch, D.R.

    2004-01-01

    We report on an ongoing study on modular Heavy Ion Fusion drivers. The modular driver is characterized by 10 to 20 nearly identical induction linacs, each carrying a single high current beam. In this scheme, the Integrated Research Experiment (IRE) can be one of the full size induction linacs. Hence, this approach offers significant advantages in terms of driver development path. For beam transport, these modules use solenoids which are capable of carrying high line charge densities, even at low energies. A new injector concept allows compression of the beam to high line densities right at the source. The final drift compression is performed in a plasma, in which the large repulsive space charge effects are neutralized. Finally, the beam is transversely compressed onto the target, using either external solenoids or current-carrying channels (in the Assisted Pinch Mode of beam propagation). We will report on progress towards a self-consistent point design from injector to target. Considerations of driver architecture, chamber environment as well as the methodology for meeting target requirements of spot size, pulse shape and symmetry will also be described. Finally, some near-term experiments to address the key scientific issues will be discussed

  17. The Modular Point Design for Heavy Ion Fusion

    International Nuclear Information System (INIS)

    Yu, S.S.; Barnard, J.J.; Briggs, R.J.; Callahan-Miller, D.; Celata, C.M.; Chao, L.; Davidson, R.; Debonnel, C.S.; Eylon, S.; Friedman, A.; Henestroza, E.; Kaganovich, I.; Kwan, J.W.; Lee, E.P.; Leitner, M.; Logan, B.G.; Meier, W.; Peterson, P.F.; Reginato, L.; Rose, D.; Roy, P.; Waldron, W.; Welch, D.R.

    2005-01-01

    We report on an ongoing study on modular Heavy Ion Fusion drivers. The modular driver is characterized by tens (∼ 20) nearly identical induction linacs, each carrying a single high current beam. In this scheme, the Integrated Research Experiment (IRE) can be one of the full size induction linacs. Hence, this approach offers significant advantages in terms of driver development path. For beam transport, these modules use solenoids which are capable of carrying high line charge densities, even at low energies. A new injector concept allows compression of the beam to high line densities right at the source. The final drift compression is performed in a plasma, in which the large repulsive space charge effects are neutralized. Finally, the beam is transversely compressed onto the target, using either external solenoids or current-carrying channels (in the Assisted Pinch Mode of beam propagation). We will report on progress towards a self-consistent point design from injector to target. Considerations of driver architecture, chamber environment as well as the methodology for meeting target requirements of spot size, pulse shape and symmetry will also be described. Finally, some near-term experiments to address the key scientific issues will be discussed

  18. Critical safety issues in the design of fusion machines

    International Nuclear Information System (INIS)

    Kramer, W.

    1991-01-01

    In the course of developing fusion machines both general safety considerations and safety assessments for the various components and systems of actual machines increase in number and become more and more coherent. This is particularly true for the NET/ITER projects where safety analysis plays an increasing role for the design of the machine. Since in a D/T tokamak the radiological hazards will be dominant basic radiological safety objectives are discussed. Critical safety issues as identified in particular by the NET/ITER community are reviewed. Subsequently, issues of major concern are considered both for normal operation and for conceivable accidents. The following accidents are considered to be crucial: Loss of cooling in plasma facing components, loss of vacuum, tritium system failure, and magnet system failure. To mitigate accident consequences a confinement concept based on passive features and multiple barriers including detritiation and filtering has to be applied. The reactor building as final barrier needs special attention to cope with both internal and external hazards. (orig.)

  19. Design of the TFTR [Tokamak Fusion Test Reactor] maintenance manipulator

    International Nuclear Information System (INIS)

    Loesser, G. D.; Heitzenroeder, P.; Bohme, G.; Selig, M.

    1987-01-01

    The Tokamak Fusion Test Reactor (TFTR) plans to generate a total of 3 x 10 21 neutrons during its deuterium-tritium run period in 1900. This will result in high levels of radiation, especially within the TFTR vacuum vessel. The maintenance manipulator's mission is to assist TFTR in meeting Princeton Plasma Physics Laboratory's personnel radiation exposure criteria and in maintaining as-low-as-reasonably-achievable principals by limiting the radiation exposure received by operating and maintenance personnel. The manipulator, which is currently being fabricated and tested by Kernforschungszentrum Karlsruhe, is designed to perform limited, but routine and necessary, functions within the TFTR vacuum torus after activation levels within the torus preclude such functions being performed by personnel. These functions include visual inspection, tile replacement, housekeeping tasks, diagnostic calibrations, and leak detection. To meet its functional objectives, the TFTR maintenance manipulator is required to be operable in TFTR's very high vacuum environment (typically 2 x 10 -8 Torr). It must also be bakeable at 150 degree C and able to withstand the radiation environment

  20. In vivo immobilization of fusion proteins on bioplastics by the novel tag BioF.

    Science.gov (United States)

    Moldes, Cristina; García, Pedro; García, José L; Prieto, María A

    2004-06-01

    A new protein immobilization and purification system has been developed based on the use of polyhydroxyalkanoates (PHAs, or bioplastics), which are biodegradable polymers accumulated as reserve granules in the cytoplasm of certain bacteria. The N-terminal domain of the PhaF phasin (a PHA-granule-associated protein) from Pseudomonas putida GPo1 was used as a polypeptide tag (BioF) to anchor fusion proteins to PHAs. This tag provides a novel way to immobilize proteins in vivo by using bioplastics as supports. The granules carrying the BioF fusion proteins can be isolated by a simple centrifugation step and used directly for some applications. Moreover, when required, a practically pure preparation of the soluble BioF fusion protein can be obtained by a mild detergent treatment of the granule. The efficiency of this system has been demonstrated by constructing two BioF fusion products, including a functional BioF-beta-galactosidase. This is the first example of an active bioplastic consisting of a biodegradable matrix carrying an active enzyme.

  1. An ER-directed fusion protein comprising a bacterial subtilisin ...

    African Journals Online (AJOL)

    nausch

    subtilase tag was fused to human interleukin 6 (IL6) and transiently expressed in Nicotiana ..... MP, tobacco mosaic virus (TMV) movement protein; TVCV-3'-NTR, TVCV-3' untranslated .... on the degradation pattern of heterologous proteins.

  2. Directed supramolecular surface assembly of SNAP-tag fusion proteins

    NARCIS (Netherlands)

    Uhlenheuer, D.A.; Wasserberg, D.; Haase, C.; Nguyen, H.; Schenkel, J.H.; Huskens, J.; Ravoo, B.J.; Jonkheijm, P.; Brunsveld, L.

    2012-01-01

    Supramolecular assembly of proteins on surfaces and vesicles was investigated by site-selective incorporation of a supramolecular guest element on proteins. Fluorescent proteins were site-selectively labeled with bisadamantane by SNAP-tag technology. The assembly of the bisadamantane functionalized

  3. Immobilization of ferrocene-modified SNAP-fusion proteins

    NARCIS (Netherlands)

    Wasserberg, D.; Uhlenheuer, D.; Neirynck, P.; Neirynck, Pauline; Cabanas Danés, Jordi; Schenkel, J.H.; Ravoo, B.J.; An, Q.; Huskens, Jurriaan; Milroy, L.G.; Brunsveld, Luc; Jonkheijm, Pascal

    2013-01-01

    The supramolecular assembly of proteins on surfaces has been investigated via the site-selective incorporation of a supramolecular moiety on proteins. To this end, fluorescent proteins have been site-selectively labeled with ferrocenes, as supramolecular guest moieties, via SNAP-tag technology. The

  4. Directed Supramolecular Surface Assembly of SNAP-tag Fusion Proteins

    NARCIS (Netherlands)

    Uhlenheuer, D.A.; Wasserberg, D.; Haase, C.; Nguyen, Hoang D.; Schenkel, J.H.; Huskens, Jurriaan; Ravoo, B.J.; Jonkheijm, Pascal; Brunsveld, Luc

    2012-01-01

    Supramolecular assembly of proteins on surfaces and vesicles was investigated by site-selective incorporation of a supramolecular guest element on proteins. Fluorescent proteins were site-selectively labeled with bisadamantane by SNAP-tag technology. The assembly of the bisadamantane functionalized

  5. Fusion energy

    International Nuclear Information System (INIS)

    Gross, R.A.

    1984-01-01

    This textbook covers the physics and technology upon which future fusion power reactors will be based. It reviews the history of fusion, reaction physics, plasma physics, heating, and confinement. Descriptions of commercial plants and design concepts are included. Topics covered include: fusion reactions and fuel resources; reaction rates; ignition, and confinement; basic plasma directory; Tokamak confinement physics; fusion technology; STARFIRE: A commercial Tokamak fusion power plant. MARS: A tandem-mirror fusion power plant; and other fusion reactor concepts

  6. Computational design of protein interactions: designing proteins that neutralize influenza by inhibiting its hemagglutinin surface protein

    Science.gov (United States)

    Fleishman, Sarel

    2012-02-01

    Molecular recognition underlies all life processes. Design of interactions not seen in nature is a test of our understanding of molecular recognition and could unlock the vast potential of subtle control over molecular interaction networks, allowing the design of novel diagnostics and therapeutics for basic and applied research. We developed the first general method for designing protein interactions. The method starts by computing a region of high affinity interactions between dismembered amino acid residues and the target surface and then identifying proteins that can harbor these residues. Designs are tested experimentally for binding the target surface and successful ones are affinity matured using yeast cell surface display. Applied to the conserved stem region of influenza hemagglutinin we designed two unrelated proteins that, following affinity maturation, bound hemagglutinin at subnanomolar dissociation constants. Co-crystal structures of hemagglutinin bound to the two designed binders were within 1Angstrom RMSd of their models, validating the accuracy of the design strategy. One of the designed proteins inhibits the conformational changes that underlie hemagglutinin's cell-invasion functions and blocks virus infectivity in cell culture, suggesting that such proteins may in future serve as diagnostics and antivirals against a wide range of pathogenic influenza strains. We have used this method to obtain experimentally validated binders of several other target proteins, demonstrating the generality of the approach. We discuss the combination of modeling and high-throughput characterization of design variants which has been key to the success of this approach, as well as how we have used the data obtained in this project to enhance our understanding of molecular recognition. References: Science 332:816 JMB, in press Protein Sci 20:753

  7. Conceptual fusion reactor designs based on the laser heat solenoid

    International Nuclear Information System (INIS)

    Steinhauer, L.C.

    1976-01-01

    The feasibility of the laser heated solenoid (LHS) as an approach to fusion and fusion-fission commercial power generation has been examined. The LHS concept is based on magnetic confinement of a long slender plasma column which is partly heated by the axially directed beam from a powerful long wavelength laser. As a pure fusion concept, the LHS configurations studied so far are characterized by fairly difficult engineering constraints, particularly on the magnet, a large laser, and a marginally acceptable system energy balance. As a fusion-fission system, however, the LHS is capable of a very attractive energy balance, has much more relaxed engineering constraints, requires a relatively modest laser, and as such holds great potential as a power generator and fissile fuel breeding scheme

  8. Trends in radiation protection: possible effects on fusion power plant design

    International Nuclear Information System (INIS)

    Eurajoki, Tapani; Frias, Manuel Pascual; Orlandi, Sergio

    2003-01-01

    Since the design of fusion power plants involves long-term issues, ranging over several decades, it is useful to try to foresee under what kind of regulations the first fusion plants are to be operated. Application of present radiological regulations and practice to a fusion power plant concept is considered. The current design phase of fusion power plants motivates the top-down dose assessment, but it is crucial to aim at bottom-up assessments to ensure radiation doses as low as reasonably achievable. Since several issues, relating both to our knowledge on radiation as well as to the practice of radiation protection, may change in the future, it is necessary to continuously follow the development in the further design of fusion power plants

  9. Potential design modifications for the High Yield Lithium Injection Fusion Energy (HYLIFE) reaction chamber

    International Nuclear Information System (INIS)

    Pitts, J.H.; Hovingh, J.; Meier, W.R.; Monsler, M.J.; Powell, E.G.; Walker, P.E.

    1979-01-01

    Generation of electric power from inertial confinement fusion requires a reaction chamber. One promising type, the High Yield Lithium Injection Fusion Energy (HYLIFE) chamber, includes a falling array of liquid lithium jets. These jets act as: (1) a renewable first wall and blanket to shield metal components from x-ray and neutron exposure, (2) a tritium breeder to replace tritium burned during the fusion process, and (3) an absorber and transfer medium for fusion energy. Over 90% of the energy produced in the reaction chamber is absorbed in the lithium jet fall. Design aspects are included

  10. Recent contributions to fusion reactor design and technology development

    International Nuclear Information System (INIS)

    1979-11-01

    The report contains a collection of 16 recent fusion technology papers on the STARFIRE Project, the study of alternate fusion fuel cycles, a maintainability study, magnet safety, neutral beam power supplies and pulsed superconducting magnets and energy transfer. This collection of papers contains contributions for Argonne National Laboratory, McDonnell Douglas Astronautics Company, General Atomic Company, The Ralph M. Parsons Company, the University of Illinois, and the University of Wisconsin. Separate abstracts are presented for each paper

  11. Inhibition of CRM1-mediated nuclear export of transcription factors by leukemogenic NUP98 fusion proteins.

    Science.gov (United States)

    Takeda, Akiko; Sarma, Nayan J; Abdul-Nabi, Anmaar M; Yaseen, Nabeel R

    2010-05-21

    NUP98 is a nucleoporin that plays complex roles in the nucleocytoplasmic trafficking of macromolecules. Rearrangements of the NUP98 gene in human leukemia result in the expression of numerous fusion oncoproteins whose effect on nucleocytoplasmic trafficking is poorly understood. The present study was undertaken to determine the effects of leukemogenic NUP98 fusion proteins on CRM1-mediated nuclear export. NUP98-HOXA9, a prototypic NUP98 fusion, inhibited the nuclear export of two known CRM1 substrates: mutated cytoplasmic nucleophosmin and HIV-1 Rev. In vitro binding assays revealed that NUP98-HOXA9 binds CRM1 through the FG repeat motif in a Ran-GTP-dependent manner similar to but stronger than the interaction between CRM1 and its export substrates. Two NUP98 fusions, NUP98-HOXA9 and NUP98-DDX10, whose fusion partners are structurally and functionally unrelated, interacted with endogenous CRM1 in myeloid cells as shown by co-immunoprecipitation. These leukemogenic NUP98 fusion proteins interacted with CRM1, Ran, and the nucleoporin NUP214 in a manner fundamentally different from that of wild-type NUP98. NUP98-HOXA9 and NUP98-DDX10 formed characteristic aggregates within the nuclei of a myeloid cell line and primary human CD34+ cells and caused aberrant localization of CRM1 to these aggregates. These NUP98 fusions caused nuclear accumulation of two transcription factors, NFAT and NFkappaB, that are regulated by CRM1-mediated export. The nuclear entrapment of NFAT and NFkappaB correlated with enhanced transcription from promoters responsive to these transcription factors. Taken together, the results suggest a new mechanism by which NUP98 fusions dysregulate transcription and cause leukemia, namely, inhibition of CRM1-mediated nuclear export with aberrant nuclear retention of transcriptional regulators.

  12. Development of a novel fluorescent imaging probe for tumor hypoxia by use of a fusion protein with oxygen-dependent degradation domain of HIF-1α

    Science.gov (United States)

    Tanaka, Shotaro; Kizaka-Kondoh, Shinae; Harada, Hiroshi; Hiraoka, Masahiro

    2007-02-01

    More malignant tumors contain more hypoxic regions. In hypoxic tumor cells, expression of a series of hypoxiaresponsive genes related to malignant phenotype such as angiogenesis and metastasis are induced. Hypoxia-inducible factor-1 (HIF-1) is a master transcriptional activator of such genes, and thus imaging of hypoxic tumor cells where HIF-1 is active, is important in cancer therapy. We have been developing PTD-ODD fusion proteins, which contain protein transduction domain (PTD) and the VHL-mediated protein destruction motif in oxygen-dependent degradation (ODD) domain of HIF-1 alpha subunit (HIF-1α). Thus PTD-ODD fusion proteins can be delivered to any tissue in vivo through PTD function and specifically stabilized in hypoxic cells through ODD function. To investigate if PTD-ODD fusion protein can be applied to construct hypoxia-specific imaging probes, we first constructed a fluorescent probe because optical imaging enable us to evaluate a probe easily, quickly and economically in a small animal. We first construct a model fusion porein PTD-ODD-EGFP-Cy5.5 named POEC, which is PTD-ODD protein fused with EGFP for in vitro imaging and stabilization of fusion protein, and conjugated with a near-infrared dye Cy5.5. This probe is designed to be degraded in normoxic cells through the function of ODD domain and followed by quick clearance of free fluorescent dye. On the other hand, this prove is stabilized in hypoxic tumor cells and thus the dye is stayed in the cells. Between normoxic and hypoxic conditions, the difference in the clearance rate of the dye will reveals suited contrast for tumor-hypoxia imaging. The optical imaging probe has not been optimized yet but the results presented here exhibit a potential of PTD-ODD fusion protein as a hypoxia-specific imaging probe.

  13. Site-directed antibody immobilization using a protein A-gold binding domain fusion protein for enhanced SPR immunosensing.

    Science.gov (United States)

    de Juan-Franco, Elena; Caruz, Antonio; Pedrajas, J R; Lechuga, Laura M

    2013-04-07

    We have implemented a novel strategy for the oriented immobilization of antibodies onto a gold surface based on the use of a fusion protein, the protein A-gold binding domain (PAG). PAG consists of a gold binding peptide (GBP) coupled to the immunoglobulin-binding domains of staphylococcal protein A. This fusion protein provides an easy and fast oriented immobilization of antibodies preserving its native structure, while leaving the antigen binding sites (Fab) freely exposed. Using this immobilization strategy, we have demonstrated the performance of the immunosensing of the human Growth Hormone by SPR. A limit of detection of 90 ng mL(-1) was obtained with an inter-chip variability lower than 7%. The comparison of this method with other strategies for the direct immobilization of antibodies over gold surfaces has showed the enhanced sensitivity provided by the PAG approach.

  14. Generation of a selectively cytotoxic fusion protein against p53 mutated cancers

    International Nuclear Information System (INIS)

    Kousparou, Christina A; Yiacoumi, Efthymia; Deonarain, Mahendra P; Epenetos, Agamemnon A

    2012-01-01

    A significant number of cancers are caused by defects in p21 causing functional defects in p21 or p53 tumour-suppressor proteins. This has led to many therapeutic approaches including restoration by gene therapy with wild-type p53 or p21 using viral or liposomal vectors, which have toxicity or side-effect limitations. We set out to develop a safer, novel fusion protein which has the ability to reconstitute cancer cell lines with active p21 by protein transduction. The fusion protein was produced from the cell-translocating peptide Antennapedia (Antp) and wild-type, full-length p21 (Antp-p21). This was expressed and refolded from E. coli and tested on a variety of cell lines and tumours (in a BALB/c nude xenograft model) with differing p21 or p53 status. Antp-p21 penetrated and killed cancer cells that do not express wild type p53 or p21. This included cells that were matched to cogenic parental cell lines. Antp-p21 killed cancer cells selectively that were malignant as a result of mutations or nuclear exclusion of the p53 and p21 genes and over-expression of MDM2. Non-specific toxicity was excluded by showing that Antp-p21 penetrated but did not kill p53- or p21- wild-type cells. Antp-p21 was not immunogenic in normal New Zealand White rabbits. Recombinant Antp peptide alone was not cytotoxic, showing that killing was due to the transduction of the p21 component of Antp-p21. Antp-p21 was shown to penetrate cancer cells engrafted in vivo and resulted in tumour eradication when administered with conventionally-used chemotherapeutic agents, which alone were unable to produce such an effect. Antp-p21 may represent a new and promising targeted therapy for patients with p53-associated cancers supporting the concept that rational design of therapies directed against specific cancer mutations will play a part in the future of medical oncology

  15. Generation of a selectively cytotoxic fusion protein against p53 mutated cancers

    Directory of Open Access Journals (Sweden)

    Kousparou Christina A

    2012-08-01

    Full Text Available Abstract Background A significant number of cancers are caused by defects in p21 causing functional defects in p21 or p53 tumour-suppressor proteins. This has led to many therapeutic approaches including restoration by gene therapy with wild-type p53 or p21 using viral or liposomal vectors, which have toxicity or side-effect limitations. We set out to develop a safer, novel fusion protein which has the ability to reconstitute cancer cell lines with active p21 by protein transduction. Methods The fusion protein was produced from the cell-translocating peptide Antennapedia (Antp and wild-type, full-length p21 (Antp-p21. This was expressed and refolded from E. coli and tested on a variety of cell lines and tumours (in a BALB/c nude xenograft model with differing p21 or p53 status. Results Antp-p21 penetrated and killed cancer cells that do not express wild type p53 or p21. This included cells that were matched to cogenic parental cell lines. Antp-p21 killed cancer cells selectively that were malignant as a result of mutations or nuclear exclusion of the p53 and p21 genes and over-expression of MDM2. Non-specific toxicity was excluded by showing that Antp-p21 penetrated but did not kill p53- or p21- wild-type cells. Antp-p21 was not immunogenic in normal New Zealand White rabbits. Recombinant Antp peptide alone was not cytotoxic, showing that killing was due to the transduction of the p21 component of Antp-p21. Antp-p21 was shown to penetrate cancer cells engrafted in vivo and resulted in tumour eradication when administered with conventionally-used chemotherapeutic agents, which alone were unable to produce such an effect. Conclusions Antp-p21 may represent a new and promising targeted therapy for patients with p53-associated cancers supporting the concept that rational design of therapies directed against specific cancer mutations will play a part in the future of medical oncology.

  16. HYLIFE-II inertial fusion energy power plant design

    International Nuclear Information System (INIS)

    Moir, R.W.

    1992-01-01

    The HYLIFE-II inertial fusion power plant design study uses a liquid fall, in the form of jets, to protect the first structural wall from neutron damage, x rays, and blast to provide a 30-y lifetime. HYLIFE-I used liquid lithium. HYLIFE-II avoids the fire hazard of lithium by using a molten salt composed of fluorine, lithium, and beryllium (Li 2 BeF 4 ) called Flibe. Access for heavy-ion beams is provided. Calculations for assumed heavy-ion beam performance show a nominal gain of 70 at 5 MJ producing 350 MJ, about 5.2 times less yield than the 1.8 GJ from a driver energy of 4.5 MJ with gain of 400 for HYLIFE-I. The nominal 1 GWe of power can be maintained by increasing the repetition rate by a factor of about 5.2, from 1.5 to 8 Hz. A higher repetition rate requires faster re-establishment of the jets after a shot, which can be accomplished in part by decreasing the jet fall height and increasing the jet flow velocity. In addition, although not adequately considered for HYLIFE-I, there is liquid splash that must be forcibly cleared because gravity is too slow, at higher repetition rates than 1 Hz. Splash removal is accomplished in the central region by oscillating jet flows. The cost of electricity is estimated to be 0.09 $/kW·h in constant 1988 dollars, about twice that of future coal and light water reactor nuclear power. The driver beam cost is about one-half the total cost, that is, a zero cost driver would give a calculated cost of electricity of 0.045 $/kWh

  17. HYLIFE-II inertial fusion energy power plant design

    International Nuclear Information System (INIS)

    Moir, R.W.

    1992-01-01

    The HYLIFE-II inertial fusion power plant design study uses a liquid fall, in the form of jets, to protect the first structural wall from neutron damage, x rays, and blast to provide a 30-y lifetime. HYLIFE-I used liquid lithium. HYLIFE-II avoids the fire hazard of lithium by using a molten salt composed of fluorine, lithium, and beryllium (Li 2 BeF 4 ) called Flibe. Access for heavy-ion beams is provided. Calculations for assumed heavy-ion beam performance show a nominal gain of 70 at 5 MJ producing 350 MJ, about 5.2 times less yield than the 1.8 Gj from a driver energy of 4.5 MJ with gain of 400 for HYLIFE-I. The nominal 1 GWe of power can be maintained by increasing the repetition rate by a factor of about 5.2, from 1.5 to 8Hz. A higher repetition rate requires faster re-establishment of the jets after a shot, which can be accomplished in part by decreasing the jet fall height and increasing the jet flow velocity. In addition, although not adequately considered for HYLIFE-I, there is liquid splash that must be forcibly cleared because gravity is too slow, at higher repetition rates than 1 Hz. Splash removal is accomplished in the central region by oscillating jet flows. The cost of electricity is estimated to be 0.09 $/kW·h in constant 1988 dollars, about twice that of future coal and light water reactor nuclear power. The driver beam cost is about one-half the total cost, that is, a zero cost driver would give a calculated cost of electricity of 0.045 $/kWh

  18. HYLIFE-II inertial confinement fusion reactor design

    International Nuclear Information System (INIS)

    Moir, R.W.

    1990-01-01

    The HYLIFE-2 inertial fusion power plant design study uses a liquid fall, in the form of jets to protect the first structural wall from neutron damage, x rays, and blast to provide a 30-y lifetime. HYLIFE-1 used liquid lithium. HYLIFE 2 avoids the fire hazard of lithium by using a molten salt composed of fluorine, lithium, and beryllium (Li 2 BeF 4 ) called Flibe. Access for heavy-ion beams is provided. Calculations for assumed heavy-ion beam performance show a nominal gain of 70 at 5 MJ producing 350 MJ, about 5.2 times less yield than the 1.8 GJ from a driver energy of 4.5 MJ with gain of 400 for HYLIFE-1. The nominal 1 GWe of power can be maintained by increasing the repetition rate by a factor of about 5.2, from 1.5 to 8 Hz. A higher repetition rate requires faster re-establishment of the jets after a shot, which can be accomplished in part by decreasing the jet fall height and increasing the jet flow velocity. Multiple chambers may be required. In addition, although not considered for HYLIFE-1, there is undoubtedly liquid splash that must be forcibly cleared because gravity is too slow, especially at high repetition rates. Splash removal can be accomplished by either pulsed or oscillating jet flows. The cost of electricity is estimated to be 0.09 $/kW·h in constant 1988 dollars, about twice that of future coal and light water reactor nuclear power. The driver beam cost is about one-half the total cost. 15 refs., 9 figs., 3 tabs

  19. HYLIFE-II inertial confinement: Fusion power plant design

    International Nuclear Information System (INIS)

    Moir, R.W.

    1990-01-01

    The HYLIFE-2 inertial fusion power plant design study uses a liquid fall, in the form of jets to protect the first structural wall from neutron damage, x rays, and blast to provide a 30-y lifetime. HYLIFE-1 used liquid lithium. HYLIFE 2 avoids the fire hazard of lithium by using a molten salt composed of fluorine, lithium, and beryllium (Li 2 BeF 4 ) called Flibe. Access for heavy-ion beams is provided. Calculations for assumed heavy-ion beam performance show a nominal gain of 70 at 5 MJ producing 350 MJ, about 5.2 times less yield than the 1.8 GJ from a driver energy of 4.5 MJ with gain of 400 for HYLIFE-1. The nominal 1 GWe of power can be maintained by increasing the repetition rate by a factor of about 5.2, from 1.5 to 8 Hz. A higher repetition rate requires faster re-establishment of the jets after a shot, which can be accomplished in part by decreasing the jet fall height and increasing the jet flow velocity. Multiple chambers may be required. In addition, although not considered for HYLIFE-1, there is undoubtedly liquid splash that must be forcibly cleared because gravity is too slow, especially at high repetition rates. Splash removal can be accomplished by either pulsed or oscillating jet flows. The cost of electricity is estimated to be 0.09 $/kW·h in constant 1988 dollars, about twice that of future coal and light water reactor nuclear power. The driver beam cost is about one-half the total cost. 16 refs., 6 figs., 2 tabs

  20. Studies to Prevent Degradation of Recombinant Fc-Fusion Protein Expressed in Mammalian Cell Line and Protein Characterization

    Directory of Open Access Journals (Sweden)

    Sanjukta Chakrabarti

    2016-06-01

    Full Text Available Clipping of recombinant proteins is a major issue in animal cell cultures. A recombinant Fc-fusion protein, VEGFR1(D1–D3-Fc expressed in CHOK1SV GS-KO cells was observed to be undergoing clippings in lab scale cultures. Partial cleaving of expressed protein initiated early on in cell culture and was observed to increase over time in culture and also on storage. In this study, a few parameters were explored in a bid to inhibit clipping in the fusion protein The effects of culture temperature, duration of culture, the addition of an anti-clumping agent, ferric citrate and use of protease inhibitor cocktail on inhibition of proteolysis of the Fc fusion were studied. Lowering of culture temperature from 37 to 30 °C alone appears to be the best solution for reducing protein degradation from the quality, cost and regulatory points of view. The obtained Fc protein was characterized and found to be in its stable folded state, exhibiting a high affinity for its ligand and also biological and functional activities.

  1. Enhanced SUMOylation of proteins containing a SUMO-interacting motif by SUMO-Ubc9 fusion

    International Nuclear Information System (INIS)

    Kim, Eui Tae; Kim, Kyeong Kyu; Matunis, Mike J.; Ahn, Jin-Hyun

    2009-01-01

    Identifying new targets for SUMO and understanding the function of protein SUMOylation are largely limited by low level of SUMOylation. It was found recently that Ubc9, the SUMO E2 conjugating enzyme, is covalently modified by SUMO at a lysine 14 in the N-terminal alpha helix, and that SUMO-modified Ubc9 has enhanced conjugation activity for certain target proteins containing a SUMO-interacting motif (SIM). Here, we show that, compared to intact Ubc9, the SUMO-Ubc9 fusion protein has higher conjugating activity for SIM-containing targets such as Sp100 and human cytomegalovirus IE2. Assays using an IE2 SIM mutant revealed the requirement of SIM for the enhanced IE2 SUMOylation by SUMO-Ubc9. In pull-down assays with cell extracts, the SUMO-Ubc9 fusion protein bound to more diverse cellular proteins and interacted with some SIM-containing proteins with higher affinities than Ubc9. Therefore, the devised SUMO-Ubc9 fusion will be useful for identifying SIM-containing SUMO targets and producing SUMO-modified proteins.

  2. Integrin αvβ1 Modulation Affects Subtype B Avian Metapneumovirus Fusion Protein-mediated Cell-Cell Fusion and Virus Infection.

    Science.gov (United States)

    Yun, Bing-Ling; Guan, Xiao-Lu; Liu, Yong-Zhen; Zhang, Yao; Wang, Yong-Qiang; Qi, Xiao-Le; Cui, Hong-Yu; Liu, Chang-Jun; Zhang, Yan-Ping; Gao, Hong-Lei; Gao, Li; Li, Kai; Gao, Yu-Long; Wang, Xiao-Mei

    2016-07-08

    Avian metapneumovirus (aMPV) fusion (F) protein mediates virus-cell membrane fusion to initiate viral infection, which requires F protein binding to its receptor(s) on the host cell surface. However, the receptor(s) for aMPV F protein is still not identified. All known subtype B aMPV (aMPV/B) F proteins contain a conserved Arg-Asp-Asp (RDD) motif, suggesting that the aMPV/B F protein may mediate membrane fusion via the binding of RDD to integrin. When blocked with integrin-specific peptides, aMPV/B F protein fusogenicity and viral replication were significantly reduced. Specifically we identified integrin αv and/or β1-mediated F protein fusogenicity and viral replication using antibody blocking, small interfering RNAs (siRNAs) knockdown, and overexpression. Additionally, overexpression of integrin αv and β1 in aMPV/B non-permissive cells conferred aMPV/B F protein binding and aMPV/B infection. When RDD was altered to RAE (Arg-Ala-Glu), aMPV/B F protein binding and fusogenic activity were profoundly impaired. These results suggest that integrin αvβ1 is a functional receptor for aMPV/B F protein-mediated membrane fusion and virus infection, which will provide new insights on the fusogenic mechanism and pathogenesis of aMPV. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  3. Integrin αvβ1 Modulation Affects Subtype B Avian Metapneumovirus Fusion Protein-mediated Cell-Cell Fusion and Virus Infection*

    Science.gov (United States)

    Yun, Bing-Ling; Guan, Xiao-Lu; Liu, Yong-Zhen; Zhang, Yao; Wang, Yong-Qiang; Qi, Xiao-Le; Cui, Hong-Yu; Liu, Chang-Jun; Zhang, Yan-Ping; Gao, Hong-Lei; Gao, Li; Li, Kai; Gao, Yu-Long; Wang, Xiao-Mei

    2016-01-01

    Avian metapneumovirus (aMPV) fusion (F) protein mediates virus-cell membrane fusion to initiate viral infection, which requires F protein binding to its receptor(s) on the host cell surface. However, the receptor(s) for aMPV F protein is still not identified. All known subtype B aMPV (aMPV/B) F proteins contain a conserved Arg-Asp-Asp (RDD) motif, suggesting that the aMPV/B F protein may mediate membrane fusion via the binding of RDD to integrin. When blocked with integrin-specific peptides, aMPV/B F protein fusogenicity and viral replication were significantly reduced. Specifically we identified integrin αv and/or β1-mediated F protein fusogenicity and viral replication using antibody blocking, small interfering RNAs (siRNAs) knockdown, and overexpression. Additionally, overexpression of integrin αv and β1 in aMPV/B non-permissive cells conferred aMPV/B F protein binding and aMPV/B infection. When RDD was altered to RAE (Arg-Ala-Glu), aMPV/B F protein binding and fusogenic activity were profoundly impaired. These results suggest that integrin αvβ1 is a functional receptor for aMPV/B F protein-mediated membrane fusion and virus infection, which will provide new insights on the fusogenic mechanism and pathogenesis of aMPV. PMID:27226547

  4. New sensitive and specific assay for human immunodeficiency virus antibodies using labeled recombinant fusion protein and time-resolved fluoroimmunoassay.

    OpenAIRE

    Siitari, H; Turunen, P; Schrimsher, J; Nunn, M

    1990-01-01

    A new, rapid method for the detection of human immunodeficiency virus type 1 (HIV-1) antibody by time-resolved fluoroimmunoassay (TR-FIA) was developed. In this assay format, microtitration strips were coated with a recombinant fusion protein, and the same protein was labeled with europium and added into the wells simultaneously with the test specimens. The recombinant fusion protein contained the HIV-1 p24 gag protein sequence that carried an insertion, near the carboxyl terminus, of a 23-am...

  5. Different regions of the newcastle disease virus fusion protein modulate pathogenicity.

    Directory of Open Access Journals (Sweden)

    Sandra Heiden

    Full Text Available Newcastle disease virus (NDV, also designated as Avian paramyxovirus type 1 (APMV-1, is the causative agent of a notifiable disease of poultry but it exhibits different pathogenicity dependent on the virus strain. The molecular basis for this variability is not fully understood. The efficiency of activation of the fusion protein (F is determined by presence or absence of a polybasic amino acid sequence at an internal proteolytic cleavage site which is a major determinant of NDV virulence. However, other determinants of pathogenicity must exist since APMV-1 of high (velogenic, intermediate (mesogenic and low (lentogenic virulence specify a polybasic F cleavage site. We aimed at elucidation of additional virulence determinants by constructing a recombinant virus that consists of a lentogenic NDV Clone 30 backbone and the F protein gene from a mesogenic pigeon paramyxovirus-1 (PPMV-1 isolate with an intracerebral pathogenicity index (ICPI of 1.1 specifying the polybasic sequence R-R-K-K-R*F motif at the cleavage site. The resulting virus was characterized by an ICPI of 0.6, indicating a lentogenic pathotype. In contrast, alteration of the cleavage site G-R-Q-G-R*L of the lentogenic Clone 30 to R-R-K-K-R*F resulted in a recombinant virus with an ICPI of 1.36 which was higher than that of parental PPMV-1. Substitution of different regions of the F protein of Clone 30 by those of PPMV-1, while maintaining the polybasic amino acid sequence at the F cleavage site, resulted in recombinant viruses with ICPIs ranging from 0.59 to 1.36 suggesting that virulence is modulated by regions of the F protein other than the polybasic cleavage site.

  6. Expression and Purification of Recombinant Human Basic Fibroblast Growth Factor Fusion Proteins and Their Uses in Human Stem Cell Culture.

    Science.gov (United States)

    Imsoonthornruksa, Sumeth; Pruksananonda, Kamthorn; Parnpai, Rangsun; Rungsiwiwut, Ruttachuk; Ketudat-Cairns, Mariena

    2015-01-01

    To reduce the cost of cytokines and growth factors in stem cell research, a simple method for the production of soluble and biological active human basic fibroblast growth factor (hbFGF) fusion protein in Escherichia coli was established. Under optimal conditions, approximately 60-80 mg of >95% pure hbFGF fusion proteins (Trx-6xHis-hbFGF and 6xHis-hbFGF) were obtained from 1 liter of culture broth. The purified hbFGF proteins, both with and without the fusion tags, were biologically active, which was confirmed by their ability to stimulate proliferation of NIH3T3 cells. The fusion proteins also have the ability to support several culture passages of undifferentiated human embryonic stem cells and induce pluripotent stem cells. This paper describes a low-cost and uncomplicated method for the production and purification of biologically active hbFGF fusion proteins. © 2015 S. Karger AG, Basel.

  7. Protein design on computers. Five new proteins: Shpilka, Grendel, Fingerclasp, Leather, and Aida.

    Science.gov (United States)

    Sander, C; Vriend, G; Bazan, F; Horovitz, A; Nakamura, H; Ribas, L; Finkelstein, A V; Lockhart, A; Merkl, R; Perry, L J

    1992-02-01

    What is the current state of the art in protein design? This question was approached in a recent two-week protein design workshop sponsored by EMBO and held at the EMBL in Heidelberg. The goals were to test available design tools and to explore new design strategies. Five novel proteins were designed: Shpilka, a sandwich of two four-stranded beta-sheets, a scaffold on which to explore variations in loop topology; Grendel, a four-helical membrane anchor, ready for fusion to water-soluble functional domains; Finger-clasp, a dimer of interdigitating beta-beta-alpha units, the simplest variant of the "handshake" structural class; Aida, an antibody binding surface intended to be specific for flavodoxin; Leather--a minimal NAD binding domain, extracted from a larger protein. Each design is available as a set of three-dimensional coordinates, the corresponding amino acid sequence and a set of analytical results. The designs are placed in the public domain for scrutiny, improvement, and possible experimental verification.

  8. Low Resolution Structure of RAR1-GST-Tag Fusion Protein in Solution

    International Nuclear Information System (INIS)

    Taube, M.; Kozak, M.; Jarmolowski, A.

    2010-01-01

    RAR1 is a protein required for resistance mediated by many R genes and function upstream of signaling pathways leading to H 2 O 2 accumulation. The structure and conformation of RAR1-GST-Tag fusion protein from barley (Hordeum vulgare) in solution was studied by the small angle scattering of synchrotron radiation. It was found that the dimer of RAR1-GST-Tag protein is characterized in solution by radius of gyration R G = 6.19 nm and maximal intramolecular vector D max = 23 nm. On the basis of the small angle scattering of synchrotron radiation SAXS data two bead models obtained by ab initio modeling are proposed. Both models show elongated conformations. We also concluded that molecules of fusion protein form: dimers in solution via interaction of GST domains. (authors)

  9. The production of antibody fragments and antibody fusion proteins by yeasts and filamentous fungi

    NARCIS (Netherlands)

    Joosten, V.; Lokman, C.; Hondel, C.A.M.J.J. van den; Punt, P.J.

    2003-01-01

    In this review we will focus on the current status and views concerning the production of antibody fragments and antibody fusion proteins by yeasts and filamentous fungi. We will focus on single-chain antibody fragment production (scFv and VHH) by these lower eukaryotes and the possible applications

  10. IgM-specific serodiagnosis of acute human cytomegalovirus infection using recombinant autologous fusion proteins

    NARCIS (Netherlands)

    Vornhagen, R; Hinderer, W; Sonneborn, HH; Bein, G; Matter, L; The, T. Hauw; Enders, G; Jahn, G; Plachter, B

    Portions of three human cytomegalovirus (HCMV) polypeptides, which were shown previously to be highly reactive with patient sera, were expressed in Escherichia coli as autologous fusion proteins. Purified recombinant polypeptides were used as antigens in enzyme linked immunosorbent assay (ELISA) and

  11. Anti-Diabetic Effects of CTB-APSL Fusion Protein in Type 2 Diabetic Mice

    Directory of Open Access Journals (Sweden)

    Yunlong Liu

    2014-03-01

    Full Text Available To determine whether cholera toxin B subunit and active peptide from shark liver (CTB-APSL fusion protein plays a role in treatment of type 2 diabetic mice, the CTB-APSL gene was cloned and expressed in silkworm (Bombyx mori baculovirus expression vector system (BEVS, then the fusion protein was orally administrated at a dose of 100 mg/kg for five weeks in diabetic mice. The results demonstrated that the oral administration of CTB-APSL fusion protein can effectively reduce the levels of both fasting blood glucose (FBG and glycosylated hemoglobin (GHb, promote insulin secretion and improve insulin resistance, significantly improve lipid metabolism, reduce triglycerides (TG, total cholesterol (TC and low density lipoprotein (LDL levels and increase high density lipoprotein (HDL levels, as well as effectively improve the inflammatory response of type 2 diabetic mice through the reduction of the levels of inflammatory cytokines tumor necrosis factor-α (TNF-α and interleukin-6 (IL-6. Histopathology shows that the fusion protein can significantly repair damaged pancreatic tissue in type 2 diabetic mice, significantly improve hepatic steatosis and hepatic cell cloudy swelling, reduce the content of lipid droplets in type 2 diabetic mice, effectively inhibit renal interstitial inflammatory cells invasion and improve renal tubular epithelial cell nucleus pyknosis, thus providing an experimental basis for the development of a new type of oral therapy for type 2 diabetes.

  12. Membrane fusion is induced by a distinct peptide sequence of the sea urchin fertilization protein bindin

    NARCIS (Netherlands)

    Ulrich, AS; Glabe, CG; Hoekstra, D

    1998-01-01

    Fertilization in the sea urchin is mediated by the membrane-associated acrosomal protein bindin, which plays a key role in the adhesion and fusion between sperm and egg. We have investigated the structure/function relationship of an 18-amino acid peptide fragment "B18," which represents the minimal

  13. Design of neutron streak camera for fusion diagnostics

    International Nuclear Information System (INIS)

    Wang, C.L.; Kalibjian, R.; Singh, M.S.

    1982-06-01

    The D-T burn time for advanced laser-fusion targets is calculated to be very short, 2 . Each fission fragment leaving the cathode generates 400 secondary electrons that are all < 20 eV. These electrons are focussed to a point with an extractor and an anode, and are then purified with an electrostatic deflector. The electron beam is streaked and detected with the standard streak camera techniques. Careful shielding is needed for x-rays from the fusion target and general background. It appears that the neutron streak camera can be a viable and unique tool for studying temporal history of fusion burns in D-T plasmas of a few keV ion temperature

  14. Expression and Purification of Neurotrophin-Elastin-Like Peptide Fusion Proteins for Neural Regeneration.

    Science.gov (United States)

    Johnson, Tamina; Koria, Piyush

    2016-04-01

    Neural injuries such as spinal cord injuries, traumatic brain injuries, or nerve transection injuries pose a major health problem. Neurotrophins such as nerve growth factor (NGF) or brain-derived neurotrophic factor (BDNF) have been shown to improve the outcome of neural injuries in several pre-clinical models, but their use in clinics is limited by the lack of a robust delivery system that enhances their bioavailability and half-life. We describe two fusion proteins comprising NGF or BDNF fused with elastin-like peptides (ELPs). The aim of this study was to investigate the biological activity of neurotrophin-ELP (N-ELP) fusion proteins via in vitro culture models. NGF and BDNF were cloned in front of an elastin-like polypeptide sequence V40C2. These proteins were expressed in bacteria as inclusion bodies. These fusion proteins underwent solubilization via 8 M urea and purification via inverse transition cycling (ITC). We measured the particle size and the effect of temperature on precipitated particles using dynamic light scattering (DLS). We used western blot analysis to confirm the specificity of NGF-ELP to tropomyosin receptor kinase A (TrkA) antibody and to confirm the specificity of BDNF-ELP to TrkB antibody. PC12 cells were used to perform a neurite outgrowth assay to determine the biological activity of NGF-ELP. Bioactivity of BDNF-ELP was ascertained via transfecting human epithelial kidney (HEK 293-T) cells to express the TrkB receptor. The proteins were successfully purified to high homogeneity by exploiting the phase transition property of ELPs and urea, which solubilize inclusion bodies. Using PC12 neurite outgrowth assay, we further demonstrated that the biological activity of NGF was retained in the fusion. Similarly, BDNF-ELP phosphorylated the TrkB receptor, suggesting the biological activity of BDNF was also retained in the fusion. We further show that owing to the phase transition property of ELPs in the fusion, these proteins self-assembled into

  15. C-E1 fusion protein synthesized by rubella virus DI RNAs maintained during serial passage

    International Nuclear Information System (INIS)

    Tzeng, W.-P.; Frey, Teryl K.

    2006-01-01

    Rubella virus (RUB) replicons are derivatives of the RUB infectious cDNA clone that retain the nonstructural open reading frame (NS-ORF) that encodes the replicase proteins but not the structural protein ORF (SP-ORF) that encodes the virion proteins. RUB defective interfering (DI) RNAs contain deletions within the SP-ORF and thus resemble replicons. DI RNAs often retain the 5' end of the capsid protein (C) gene that has been shown to modulate virus-specific RNA synthesis. However, when replicons either with or without the C gene were passaged serially in the presence of wt RUB as a source of the virion proteins, it was found that neither replicon was maintained and DI RNAs were generated. The majority DI RNA species contained in-frame deletions in the SP-ORF leading to a fusion between the 5' end of the C gene and the 3' end of the E1 glycoprotein gene. DI infectious cDNA clones were constructed and transcripts from these DI infectious cDNA clones were maintained during serial passage with wt RUB. The C-E1 fusion protein encoded by the DI RNAs was synthesized and was required for maintenance of the DI RNA during serial passage. This is the first report of a functional novel gene product resulting from deletion during DI RNA generation. Thus far, the role of the C-E1 fusion protein in maintenance of DI RNAs during serial passage remained elusive as it was found that the fusion protein diminished rather than enhanced DI RNA synthesis and was not incorporated into virus particles

  16. Conceptual design of the Purdue compact torus/passive liner fusion reactor

    International Nuclear Information System (INIS)

    Terry, W.K.

    1981-01-01

    This proposal describes a program for the conceptual development of a novel fusion reactor design, the Purdue Compact Torus/Passive Liner Reactor. The key features of the concept are described and a comparison is made with a conventional tokamak

  17. Peptide-Based Membrane Fusion Inhibitors Targeting HCoV-229E Spike Protein HR1 and HR2 Domains

    Directory of Open Access Journals (Sweden)

    Shuai Xia

    2018-02-01

    Full Text Available Human coronavirus 229E (HCoV-229E infection in infants, elderly people, and immunocompromised patients can cause severe disease, thus calling for the development of effective and safe therapeutics to treat it. Here we reported the design, synthesis and characterization of two peptide-based membrane fusion inhibitors targeting HCoV-229E spike protein heptad repeat 1 (HR1 and heptad repeat 2 (HR2 domains, 229E-HR1P and 229E-HR2P, respectively. We found that 229E-HR1P and 229E-HR2P could interact to form a stable six-helix bundle and inhibit HCoV-229E spike protein-mediated cell-cell fusion with IC50 of 5.7 and 0.3 µM, respectively. 229E-HR2P effectively inhibited pseudotyped and live HCoV-229E infection with IC50 of 0.5 and 1.7 µM, respectively. In a mouse model, 229E-HR2P administered intranasally could widely distribute in the upper and lower respiratory tracts and maintain its fusion-inhibitory activity. Therefore, 229E-HR2P is a promising candidate for further development as an antiviral agent for the treatment and prevention of HCoV-229E infection.

  18. Nanoscale organization of {beta}{sub 2}-adrenergic receptor-Venus fusion protein domains on the surface of mammalian cells

    Energy Technology Data Exchange (ETDEWEB)

    Vobornik, Dusan; Rouleau, Yanouchka; Haley, Jennifer [Steacie Institute for Molecular Sciences, National Research Council Canada, Ottawa, ON, Canada K1A 0R6 (Canada); Bani-Yaghoub, Mahmud [Institute for Biological Sciences, National Research Council Canada, Ottawa, ON, Canada K1A 0R6 (Canada); Taylor, Rod [Steacie Institute for Molecular Sciences, National Research Council Canada, Ottawa, ON, Canada K1A 0R6 (Canada); Johnston, Linda J., E-mail: Linda.Johnston@nrc-cnrc.gc.ca [Steacie Institute for Molecular Sciences, National Research Council Canada, Ottawa, ON, Canada K1A 0R6 (Canada); Pezacki, John Paul, E-mail: John.Pezacki@nrc-cnrc.gc.ca [Steacie Institute for Molecular Sciences, National Research Council Canada, Ottawa, ON, Canada K1A 0R6 (Canada)

    2009-04-24

    Adrenergic receptors are a key component of nanoscale multiprotein complexes that are responsible for controlling the beat rate in a mammalian heart. We demonstrate the ability of near-field scanning optical microscopy (NSOM) to visualize {beta}{sub 2}-adrenergic receptors ({beta}{sub 2}AR) fused to the GFP analogue Venus at the nanoscale on HEK293 cells. The expression of the {beta}{sub 2}AR-Venus fusion protein was tightly controlled using a tetracycline-induced promoter. Both the size and density of the observed nanoscale domains are dependent on the level of induction and thus the level of protein expression. At concentrations between 100 and 700 ng/ml of inducer doxycycline, the size of domains containing the {beta}{sub 2}AR-Venus fusion protein appears to remain roughly constant, but the number of domains per cell increase. At 700 ng/ml doxycycline the functional receptors are organized into domains with an average diameter of 150 nm with a density similar to that observed for the native protein on primary murine cells. By contrast, larger micron-sized domains of {beta}{sub 2}AR are observed in the membrane of the HEK293 cells that stably overexpress {beta}{sub 2}AR-GFP and {beta}{sub 2}AR-eYFP. We conclude that precise chemical control of gene expression is highly advantageous for the use {beta}{sub 2}AR-Venus fusion proteins as models for {beta}{sub 2}AR function. These observations are critical for designing future cell models and assays based on {beta}{sub 2}AR, since the receptor biology is consistent with a relatively low density of nanoscale receptor domains.

  19. Engineering design of a fusion test reactor (FTR) and fusion engineering research facility (FERF) based on a toroidal theta pinch

    International Nuclear Information System (INIS)

    Abdou, M.; Burke, R.J.; Dauzvardis, P.V.; Foss, M.; Gerstl, S.A.W.; Maroni, V.A.; Pierce, A.W.; Turner, A.F.; Krakowski, R.A.; Linford, R.K.; Oliphant, T.A.; Ribe, F.L.; Thomassen, K.I.

    1975-01-01

    This paper describes two advanced toroidal theta-pinch devices which are being proposed for future construction. The Fusion Test Reactor (FTR) is being designed to produce thermonuclear energy (at 20 MeV/neutron) equal to the maximum plasma energy (Q=1) and to demonstrate α-particle heating. The Fusion Engineering and Research Facility (FERF) is being designed to test materials in a fusion environment where the average 14-MeV neutron flux from the plasma is greater than or of the order of 5.10 13 n/cm 2 .s over large surface areas. These devices employ the staged theta-pinch principle where the heating is accomplished by rapid (about 0.1 μs) implosion and expansion followed by a slow compression of the plasma. The rapid implosion injects as much heat as possible at as large a plasma radious as possible so that the plasma remains stable even after further compression. The final compression to ignition requires the transfer of a large amount of magnetic energy which implies a long transfer time (about 1 ms) for realistic voltages in the driving circuit. Throughout the heating and burn cycle the plasma must remain in equilibrium and stable to the dominant MHD-modes. A sufficiently large plasma radius guarantees stability against the m = 1 modes. These equilibrium and stability conditions and the requirements on thermonuclear burn determine the design parameters for either machine. The design parameters must also be consistent with economic limitations and technological feasibility of components. In addition to these requirements, the FERF must provide a steady and reliable source of fusion neutrons. (author)

  20. Protein Design Using Unnatural Amino Acids

    Science.gov (United States)

    Bilgiçer, Basar; Kumar, Krishna

    2003-11-01

    With the increasing availability of whole organism genome sequences, understanding protein structure and function is of capital importance. Recent developments in the methodology of incorporation of unnatural amino acids into proteins allow the exploration of proteins at a very detailed level. Furthermore, de novo design of novel protein structures and function is feasible with unprecedented sophistication. Using examples from the literature, this article describes the available methods for unnatural amino acid incorporation and highlights some recent applications including the design of hyperstable protein folds.

  1. The small G-proteins Rac1 and Cdc42 are essential for myoblast fusion in the mouse

    DEFF Research Database (Denmark)

    Vasyutina, Elena; Martarelli, Benedetta; Brakebusch, Cord

    2009-01-01

    Rac1 and Cdc42 are small G-proteins that regulate actin dynamics and affect plasma membrane protrusion and vesicle traffic. We used conditional mutagenesis in mice to demonstrate that Rac1 and Cdc42 are essential for myoblast fusion in vivo and in vitro. The deficit in fusion of Rac1 or Cdc42 mut...... genetic analysis demonstrates thus that the function of Rac in myoblast fusion is evolutionarily conserved from insects to mammals and that Cdc42, a molecule hitherto not implicated in myoblast fusion, is essential for the fusion of murine myoblasts....

  2. Myristylation of gag-onc fusion proteins in mammalian transforming retroviruses

    International Nuclear Information System (INIS)

    Schultz, A.; Oroszlan, S.

    1984-01-01

    Four cell lines producing transforming proteins encoded by three mammalian oncogenes (fes, abl, and ras) were investigated for incorporation of [ 3 H]myristate into gag-onc fusion proteins. Using 5-min pulse-labelings, fusion proteins of Abelson murine leukemia virus, Gardner-Arnstein strain of feline sarcoma virus (FeSV), and Snyder-Theilen strain of FeSV were shown to be myristylated. In a 4-hr pulse, p29gag-ras of rat sarcoma virus (RaSV) was also shown to incorporate radiolabel. The fatty acid was recovered from this labeled protein by acid hydrolysis, and identified by reverse-phase thin-layer chromatography to be [ 3 H]myristic acid. The results indicate that substitution of viral gag sequences by cellular oncogene sequences does not abolish their ability to become post-translationally modified by this long chain fatty acid. It is assumed that in the fusion proteins the myristyl moiety is linked through an amide linkage to the amino-terminal glycine as previously found for several retroviral gag precursor polyproteins. The possible role of myristylation of transforming proteins is discussed

  3. Myristylation of gag-onc fusion proteins in mammalian transforming retroviruses

    Energy Technology Data Exchange (ETDEWEB)

    Schultz, A.; Oroszlan, S.

    1984-03-01

    Four cell lines producing transforming proteins encoded by three mammalian oncogenes (fes, abl, and ras) were investigated for incorporation of (/sup 3/H)myristate into gag-onc fusion proteins. Using 5-min pulse-labelings, fusion proteins of Abelson murine leukemia virus, Gardner-Arnstein strain of feline sarcoma virus (FeSV), and Snyder-Theilen strain of FeSV were shown to be myristylated. In a 4-hr pulse, p29gag-ras of rat sarcoma virus (RaSV) was also shown to incorporate radiolabel. The fatty acid was recovered from this labeled protein by acid hydrolysis, and identified by reverse-phase thin-layer chromatography to be (/sup 3/H)myristic acid. The results indicate that substitution of viral gag sequences by cellular oncogene sequences does not abolish their ability to become post-translationally modified by this long chain fatty acid. It is assumed that in the fusion proteins the myristyl moiety is linked through an amide linkage to the amino-terminal glycine as previously found for several retroviral gag precursor polyproteins. The possible role of myristylation of transforming proteins is discussed.

  4. Safety and environmental aspects of the HYLIFE-II and ARIES fusion reactor designs

    International Nuclear Information System (INIS)

    Dolan, T.J.; Longhurst, G.R.; Herring, J.S.

    1993-01-01

    The HYLIFE-II inertial confinement fusion reactor design uses jets of Flibe molten salt to protect the blast chamber walls and to breed tritium. It has a low tritium inventory and effective tritium removal. The issue with this design is not one of safety but of economics. The ARIES reactor designs have safety concerns associated with fires. These reactors designs are described

  5. A Visual Language for Protein Design

    KAUST Repository

    Cox, Robert Sidney

    2017-02-08

    As protein engineering becomes more sophisticated, practitioners increasingly need to share diagrams for communicating protein designs. To this end, we present a draft visual language, Protein Language, that describes the high-level architecture of an engineered protein with easy-to draw glyphs, intended to be compatible with other biological diagram languages such as SBOL Visual and SBGN. Protein Language consists of glyphs for representing important features (e.g., globular domains, recognition and localization sequences, sites of covalent modification, cleavage and catalysis), rules for composing these glyphs to represent complex architectures, and rules constraining the scaling and styling of diagrams. To support Protein Language we have implemented an extensible web-based software diagram tool, Protein Designer, that uses Protein Language in a

  6. A Visual Language for Protein Design

    KAUST Repository

    Cox, Robert Sidney; McLaughlin, James Alastair; Grunberg, Raik; Beal, Jacob; Wipat, Anil; Sauro, Herbert M.

    2017-01-01

    As protein engineering becomes more sophisticated, practitioners increasingly need to share diagrams for communicating protein designs. To this end, we present a draft visual language, Protein Language, that describes the high-level architecture of an engineered protein with easy-to draw glyphs, intended to be compatible with other biological diagram languages such as SBOL Visual and SBGN. Protein Language consists of glyphs for representing important features (e.g., globular domains, recognition and localization sequences, sites of covalent modification, cleavage and catalysis), rules for composing these glyphs to represent complex architectures, and rules constraining the scaling and styling of diagrams. To support Protein Language we have implemented an extensible web-based software diagram tool, Protein Designer, that uses Protein Language in a

  7. Systems-design and energy-balance considerations for impact fusion

    International Nuclear Information System (INIS)

    Krakowski, R.A.; Miller, R.L.

    1979-01-01

    Areas of concern and potential problems for impact fusion are qualitatively considered within an overall systems context. A parametric and qualitative description of the general energy balance and systems considerations for an Impact Fusion Reactor (IFR) design is discussed. Reactor systems design considerations for an IFR are presented. An attempt to assess the IFR viability is made based on highly simplified but limiting projectile-target energy balances and thermonuclear burn models

  8. Spatiotemporal dynamics of membrane remodeling and fusion proteins during endocytic transport.

    Science.gov (United States)

    Arlt, Henning; Auffarth, Kathrin; Kurre, Rainer; Lisse, Dominik; Piehler, Jacob; Ungermann, Christian

    2015-04-01

    Organelles of the endolysosomal system undergo multiple fission and fusion events to combine sorting of selected proteins to the vacuole with endosomal recycling. This sorting requires a consecutive remodeling of the organelle surface in the course of endosomal maturation. Here we dissect the remodeling and fusion machinery on endosomes during the process of endocytosis. We traced selected GFP-tagged endosomal proteins relative to exogenously added fluorescently labeled α-factor on its way from the plasma membrane to the vacuole. Our data reveal that the machinery of endosomal fusion and ESCRT proteins has similar temporal localization on endosomes, whereas they precede the retromer cargo recognition complex. Neither deletion of retromer nor the fusion machinery with the vacuole affects this maturation process, although the kinetics seems to be delayed due to ESCRT deletion. Of importance, in strains lacking the active Rab7-like Ypt7 or the vacuolar SNARE fusion machinery, α-factor still proceeds to late endosomes with the same kinetics. This indicates that endosomal maturation is mainly controlled by the early endosomal fusion and remodeling machinery but not the downstream Rab Ypt7 or the SNARE machinery. Our data thus provide important further understanding of endosomal biogenesis in the context of cargo sorting. © 2015 Arlt et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

  9. Investigation of different cage designs and mechano-regulation algorithms in the lumbar interbody fusion process - a finite element analysis.

    Science.gov (United States)

    Postigo, Sergio; Schmidt, Hendrik; Rohlmann, Antonius; Putzier, Michael; Simón, Antonio; Duda, Georg; Checa, Sara

    2014-04-11

    Lumbar interbody fusion cages are commonly used to treat painful spinal degeneration and instability by achieving bony fusion. Many different cage designs exist, however the effect of cage morphology and material properties on the fusion process remains largely unknown. This finite element model study aims to investigate the influence of different cage designs on bone fusion using two mechano-regulation algorithms of tissue formation. It could be observed that different cages play a distinct key role in the mechanical conditions within the fusion region and therefore regulate the time course of the fusion process. Copyright © 2014 Elsevier Ltd. All rights reserved.

  10. Thermal-hydraulic analysis of low activity fusion blanket designs

    International Nuclear Information System (INIS)

    Fillo, J.A.; Powell, J.; Yu, W.S.

    1977-01-01

    The heat transfer aspects of fusion blankets are considered where: (a) conduction and (b) boiling and condensation are the dominant heat transfer mechanisms. In some cases, unique heat transfer problems arise and additional heat transfer data and analyses may be required

  11. Tokamak Fusion Test Reactor. Final conceptual design report

    International Nuclear Information System (INIS)

    1976-02-01

    The TFTR is the first U.S. magnetic confinement device planned to demonstrate the fusion of D-T at reactor power levels. This report addresses the physics objectives and the engineering goals of the TFTR project. Technical, cost, and schedule aspects of the project are included

  12. Engineering of a parainfluenza virus type 5 fusion protein (PIV-5 F): development of an autonomous and hyperfusogenic protein by a combinational mutagenesis approach.

    Science.gov (United States)

    Terrier, O; Durupt, F; Cartet, G; Thomas, L; Lina, B; Rosa-Calatrava, M

    2009-12-01

    The entry of enveloped viruses into host cells is accomplished by fusion of the viral envelope with the target cell membrane. For the paramyxovirus parainfluenza virus type 5 (PIV-5), this fusion involves an attachment protein (HN) and a class I viral fusion protein (F). We investigated the effect of 20 different combinations of 12 amino-acid substitutions within functional domains of the PIV-5 F glycoprotein, by performing cell surface expression measurements, quantitative fusion and syncytia assays. We found that combinations of mutations conferring an autonomous phenotype with mutations leading to an increased fusion activity were compatible and generated functional PIV-5 F proteins. The addition of mutations in the heptad-repeat domains led to both autonomous and hyperfusogenic phenotypes, despite the low cell surface expression of the corresponding mutants. Such engineering approach may prove useful not only for deciphering the fundamental mechanism behind viral-mediated membrane fusion but also in the development of potential therapeutic applications.

  13. Novel fusion protein approach for efficient high-throughput screening of small molecule-mediating protein-protein interactions in cells and living animals.

    Science.gov (United States)

    Paulmurugan, Ramasamy; Gambhir, Sanjiv S

    2005-08-15

    Networks of protein interactions execute many different intracellular pathways. Small molecules either synthesized within the cell or obtained from the external environment mediate many of these protein-protein interactions. The study of these small molecule-mediated protein-protein interactions is important in understanding abnormal signal transduction pathways in a variety of disorders, as well as in optimizing the process of drug development and validation. In this study, we evaluated the rapamycin-mediated interaction of the human proteins FK506-binding protein (FKBP12) rapamycin-binding domain (FRB) and FKBP12 by constructing a fusion of these proteins with a split-Renilla luciferase or a split enhanced green fluorescent protein (split-EGFP) such that complementation of the reporter fragments occurs in the presence of rapamycin. Different linker peptides in the fusion protein were evaluated for the efficient maintenance of complemented reporter activity. This system was studied in both cell culture and xenografts in living animals. We found that peptide linkers with two or four EAAAR repeat showed higher protein-protein interaction-mediated signal with lower background signal compared with having no linker or linkers with amino acid sequences GGGGSGGGGS, ACGSLSCGSF, and ACGSLSCGSFACGSLSCGSF. A 9 +/- 2-fold increase in signal intensity both in cell culture and in living mice was seen compared with a system that expresses both reporter fragments and the interacting proteins separately. In this fusion system, rapamycin induced heterodimerization of the FRB and FKBP12 moieties occurred rapidly even at very lower concentrations (0.00001 nmol/L) of rapamycin. For a similar fusion system employing split-EGFP, flow cytometry analysis showed significant level of rapamycin-induced complementation.

  14. Single-step affinity and cost-effective purification of recombinant proteins using the Sepharose-binding lectin-tag from the mushroom Laetiporus sulphureus as fusion partner.

    Science.gov (United States)

    Li, Xiao-Jing; Liu, Jin-Ling; Gao, Dong-Sheng; Wan, Wen-Yan; Yang, Xia; Li, Yong-Tao; Chang, Hong-Tao; Chen, Lu; Wang, Chuan-Qing; Zhao, Jun

    2016-03-01

    Previous research showed that a lectin from the mushroom Laetiporus sulphureus, designed LSL, bound to Sepharose and could be eluted by lactose. In this study, by taking advantage of the strong affinity of LSL-tag for Sepharose, we developed a single-step purification method for LSL-tagged fusion proteins. We utilized unmodified Sepharose-4B as a specific adsorbent and 0.2 M lactose solution as an elution buffer. Fusion proteins of LSL-tag and porcine circovirus capsid protein, designated LSL-Cap was recovered with purity of 90 ± 4%, and yield of 87 ± 3% from crude extract of recombinant Escherichia coli. To enable the remove of LSL-tag, tobacco etch virus (TEV) protease recognition sequence was placed downstream of LSL-tag in the expression vector, and LSL-tagged TEV protease, designated LSL-TEV, was also expressed in E. coli., and was recovered with purity of 82 ± 5%, and yield of 85 ± 2% from crude extract of recombinant E. coli. After digestion of LSL-tagged recombinant proteins with LSL-TEV, the LSL tag and LSL-TEV can be easily removed by passing the digested products through the Sepharose column. It is of worthy noting that the Sepharose can be reused after washing with PBS. The LSL affinity purification method enables rapid and inexpensive purification of LSL-tagged fusion proteins and scale-up production of native proteins. Copyright © 2015 Elsevier Inc. All rights reserved.

  15. Characterization of a novel fusion protein from IpaB and IpaD of Shigella spp. and its potential as a pan-Shigella vaccine.

    Science.gov (United States)

    Martinez-Becerra, Francisco J; Chen, Xiaotong; Dickenson, Nicholas E; Choudhari, Shyamal P; Harrison, Kelly; Clements, John D; Picking, William D; Van De Verg, Lillian L; Walker, Richard I; Picking, Wendy L

    2013-12-01

    Shigellosis is an important disease in the developing world, where about 90 million people become infected with Shigella spp. each year. We previously demonstrated that the type three secretion apparatus (T3SA) proteins IpaB and IpaD are protective antigens in the mouse lethal pulmonary model. In order to simplify vaccine formulation and process development, we have evaluated a vaccine design that incorporates both of these previously tested Shigella antigens into a single polypeptide chain. To determine if this fusion protein (DB fusion) retains the antigenic and protective capacities of IpaB and IpaD, we immunized mice with the DB fusion and compared the immune response to that elicited by the IpaB/IpaD combination vaccine. Purification of the DB fusion required coexpression with IpgC, the IpaB chaperone, and after purification it maintained the highly α-helical characteristics of IpaB and IpaD. The DB fusion also induced comparable immune responses and retained the ability to protect mice against Shigella flexneri and S. sonnei in the lethal pulmonary challenge. It also offered limited protection against S. dysenteriae challenge. Our results show the feasibility of generating a protective Shigella vaccine comprised of the DB fusion.

  16. Characterization of a Novel Fusion Protein from IpaB and IpaD of Shigella spp. and Its Potential as a Pan-Shigella Vaccine

    Science.gov (United States)

    Martinez-Becerra, Francisco J.; Chen, Xiaotong; Dickenson, Nicholas E.; Choudhari, Shyamal P.; Harrison, Kelly; Clements, John D.; Picking, William D.; Van De Verg, Lillian L.; Walker, Richard I.

    2013-01-01

    Shigellosis is an important disease in the developing world, where about 90 million people become infected with Shigella spp. each year. We previously demonstrated that the type three secretion apparatus (T3SA) proteins IpaB and IpaD are protective antigens in the mouse lethal pulmonary model. In order to simplify vaccine formulation and process development, we have evaluated a vaccine design that incorporates both of these previously tested Shigella antigens into a single polypeptide chain. To determine if this fusion protein (DB fusion) retains the antigenic and protective capacities of IpaB and IpaD, we immunized mice with the DB fusion and compared the immune response to that elicited by the IpaB/IpaD combination vaccine. Purification of the DB fusion required coexpression with IpgC, the IpaB chaperone, and after purification it maintained the highly α-helical characteristics of IpaB and IpaD. The DB fusion also induced comparable immune responses and retained the ability to protect mice against Shigella flexneri and S. sonnei in the lethal pulmonary challenge. It also offered limited protection against S. dysenteriae challenge. Our results show the feasibility of generating a protective Shigella vaccine comprised of the DB fusion. PMID:24060976

  17. Autographa californica multiple nucleopolyhedrovirus GP64 protein: Analysis of domain I and V amino acid interactions and membrane fusion activity

    Energy Technology Data Exchange (ETDEWEB)

    Yu, Qianlong [State Key Laboratory of Crop Stress Biology for Arid Areas, Key Laboratory of Northwest Loess Plateau Crop Pest Management of Ministry of Agriculture, College of Plant Protection, Northwest A& F University, Yangling, Shaanxi 712100 (China); Blissard, Gary W. [Boyce Thompson Institute, Cornell University, Ithaca, NY 14853, United State (United States); Liu, Tong-Xian [State Key Laboratory of Crop Stress Biology for Arid Areas, Key Laboratory of Northwest Loess Plateau Crop Pest Management of Ministry of Agriculture, College of Plant Protection, Northwest A& F University, Yangling, Shaanxi 712100 (China); Li, Zhaofei, E-mail: zhaofeili73@outlook.com [State Key Laboratory of Crop Stress Biology for Arid Areas, Key Laboratory of Northwest Loess Plateau Crop Pest Management of Ministry of Agriculture, College of Plant Protection, Northwest A& F University, Yangling, Shaanxi 712100 (China)

    2016-01-15

    The Autographa californica multiple nucleopolyhedrovirus GP64 is a class III viral fusion protein. Although the post-fusion structure of GP64 has been solved, its pre-fusion structure and the detailed mechanism of conformational change are unknown. In GP64, domain V is predicted to interact with two domain I segments that flank fusion loop 2. To evaluate the significance of the amino acids involved in these interactions, we examined 24 amino acid positions that represent interacting and conserved residues within domains I and V. In several cases, substitution of a single amino acid involved in a predicted interaction disrupted membrane fusion activity, but no single amino acid pair appears to be absolutely required. We identified 4 critical residues in domain V (G438, W439, T452, and T456) that are important for membrane fusion, and two residues (G438 and W439) that appear to be important for formation or stability of the pre-fusion conformation of GP64. - Highlights: • The baculovirus envelope glycoprotein GP64 is a class III viral fusion protein. • The detailed mechanism of conformational change of GP64 is unknown. • We analyzed 24 positions that might stabilize the post-fusion structure of GP64. • We identified 4 residues in domain V that were critical for membrane fusion. • Two residues are critical for formation of the pre-fusion conformation of GP64.

  18. Design of a fusion reaction-history measurement system with high temporal resolution

    International Nuclear Information System (INIS)

    Peng Xiaoshi; Wang Feng; Liu Shenye; Jiang Xiaohua; Tang Qi

    2010-01-01

    In order to accurately measure the history of fusion reaction for experimental study of inertial confinement fusion, we advance the design of a fusion reaction-history measurement system with high temporal resolution. The diagnostic system is composed of plastic scintillator and nose cone, an optical imaging system and the system of optic streak camera. Analyzing the capability of the system indicated that the instrument measured fusion reaction history at temporal resolution as low as 55ps and 40ps correspond to 2.45MeV DD neutrons and 14.03MeV DT neutrons. The instrument is able to measure the fusion reaction history at yields 1.5 x 10 9 DD neutrons, about 4 x 10 8 DT neutrons are required for a similar quality signal. (authors)

  19. Conceptual design and neutronics analyses of a fusion reactor blanket simulation facility

    International Nuclear Information System (INIS)

    Beller, D.E.; Ott, K.O.; Terry, W.K.

    1987-01-01

    A new conceptual design of a fusion reactor blanket simulation facility has been developed. This design follows the principles that have been successfully employed in the Purdue Fast Breeder Blanket Facility (FBBF), where experiments have resulted in the discovery of substantial deficiencies in neutronics predictions. With this design, discrepancies between calculation and experimental data can be nearly fully attributed to calculation methods because design deficiencies that could affect results are insignificant. The conceptual design of this FBBF analog, the Fusion Reactor Blanket Facility, is presented

  20. Characterization and Oral Delivery of Proinsulin-Transferrin Fusion Protein Expressed Using ExpressTec

    Directory of Open Access Journals (Sweden)

    Yu-Sheng Chen

    2018-01-01

    Full Text Available Proinsulin-transferrin fusion protein (ProINS-Tf has been designed and successfully expressed from the mammalian HEK293 cells (HEK-ProINS-Tf. It was found that HEK-ProINS-Tf could be converted into an activated form in the liver. Furthermore, HEK-ProINS-Tf was demonstrated as an extra-long acting insulin analogue with liver-specific insulin action in streptozotocin (STZ-induced type 1 diabetic mice. However, due to the low production yield from transfected HEK293 cells, there are other interesting features, including the oral bioavailability, which have not been fully explored and characterized. To improve the protein production yield, an alternative protein expression system, ExpressTec using transgenic rice (Oryza sativa L., was used. The intact and active rice-derived ProINS-Tf (ExpressTec-ProINS-Tf was successfully expressed from the transgenic rice expression system. Our results suggested that, although the insulin-like bioactivity of ExpressTec-ProINS-Tf was slightly lower in vitro, its potency of in vivo blood glucose control was considerably stronger than that of HEK-ProINS-Tf. The oral delivery studies in type 1 diabetic mice demonstrated a prolonged control of blood glucose to near-normal levels after oral administration of ExpressTec-ProINS-Tf. Results in this report suggest that ExpressTec-ProINS-Tf is a promising insulin analog with advantages including low cost, prolonged and liver targeting effects, and most importantly, oral bioactivity.

  1. [Construction of cTnC-linker-TnI (P) Genes, Expression of Fusion Protein and Preparation of Lyophilized Protein].

    Science.gov (United States)

    Song, Xiaoli; Liu, Xiaoyun; Cai, Lei; Wu, Jianwei; Wang, Jihua

    2015-12-01

    In order to construct and express human cardiac troponin C-linker-troponin I(P) [ cTnC-linker-TnI(P)] fusion protein, detect its activity and prepare lyophilized protein, we searched the CDs of human cTnC and cTnI from GenBank, synthesized cTnC and cTnI(30-110aa) into cloning vector by a short DNA sequence coding for 15 neutral amino acid residues. pCold I-cTnC-linker-TnI(P) was constructed and transformed into E. coli BL21(DE3). Then, cTnC-linker-TnI(P) fusion protein was induced by isopropyl-β-D-thiogalactopyranoside (IPTG). Soluable expression of cTnC-linker-TnI(P) in prokaryotic system was successfully obtained. The fusion protein was purified by Ni²⁺ Sepharose 6 Fast Flow affinity chromatography with over 95% purity and prepared into lyophilized protein. The activity of purified cTnC-linker-TnI(P) and its lyophilized protein were detected by Wondfo Finecare™ cTnI Test. Lyophilized protein of cTnC-linker-TnI(P) was stable for 10 or more days at 37 °C and 4 or more months at 25 °C and 4 °C. The expression system established in this research is feasible and efficient. Lyophilized protein is stable enough to be provided as biological raw materials for further research.

  2. Fusion protein based on Grb2-SH2 domain for cancer therapy

    International Nuclear Information System (INIS)

    Saito, Yuriko; Furukawa, Takako; Arano, Yasushi; Fujibayashi, Yasuhisa; Saga, Tsuneo

    2010-01-01

    Research highlights: → Grb2 mediates EGFR signaling through binding to phosphorylate EGFR with SH2 domain. → We generated fusion proteins containing 1 or 2 SH2 domains of Grb2 added with TAT. → The one with 2 SH2 domains (TSSF) interfered ERK phosphorylation. → TSSF significantly delayed the growth of EGFR overexpressing tumor in a mouse model. -- Abstract: Epidermal growth factor receptor (EGFR) is one of the very attractive targets for cancer therapy. In this study, we generated fusion proteins containing one or two Src-homology 2 (SH2) domains of growth factor receptor bound protein 2 (Grb2), which bind to phosphorylated EGFR, added with HIV-1 transactivating transcription for cell membrane penetration (termed TSF and TSSF, respectively). We examined if they can interfere Grb2-mediated signaling pathway and suppress tumor growth as expected from the lack of SH3 domain, which is necessary to intermediate EGFR-Grb2 cell signaling, in the fusion proteins. The transduction efficiency of TSSF was similar to that of TSF, but the binding activity of TSSF to EGFR was higher than that of TSF. Treatment of EGFR-overexpressing cells showed that TSSF decreased p42-ERK phosphorylation, while TSF did not. Both the proteins delayed cell growth but did not induce cell death in culture. TSSF also significantly suppressed tumor growth in vivo under consecutive administration. In conclusion, TSSF showed an ability to inhibit EGFR-Grb2 signaling and could have a potential to treat EGFR-activated cancer.

  3. Domain fusion analysis by applying relational algebra to protein sequence and domain databases.

    Science.gov (United States)

    Truong, Kevin; Ikura, Mitsuhiko

    2003-05-06

    Domain fusion analysis is a useful method to predict functionally linked proteins that may be involved in direct protein-protein interactions or in the same metabolic or signaling pathway. As separate domain databases like BLOCKS, PROSITE, Pfam, SMART, PRINTS-S, ProDom, TIGRFAMs, and amalgamated domain databases like InterPro continue to grow in size and quality, a computational method to perform domain fusion analysis that leverages on these efforts will become increasingly powerful. This paper proposes a computational method employing relational algebra to find domain fusions in protein sequence databases. The feasibility of this method was illustrated on the SWISS-PROT+TrEMBL sequence database using domain predictions from the Pfam HMM (hidden Markov model) database. We identified 235 and 189 putative functionally linked protein partners in H. sapiens and S. cerevisiae, respectively. From scientific literature, we were able to confirm many of these functional linkages, while the remainder offer testable experimental hypothesis. Results can be viewed at http://calcium.uhnres.utoronto.ca/pi. As the analysis can be computed quickly on any relational database that supports standard SQL (structured query language), it can be dynamically updated along with the sequence and domain databases, thereby improving the quality of predictions over time.

  4. Structural characterization of the fusion core in syncytin, envelope protein of human endogenous retrovirus family W

    International Nuclear Information System (INIS)

    Gong Rui; Peng Xiaoxue; Kang Shuli; Feng Huixing; Huang Jianying; Zhang Wentao; Lin Donghai; Tien Po; Xiao Gengfu

    2005-01-01

    Syncytin is a captive retroviral envelope protein, possibly involved in the formation of the placental syncytiotrophoblast layer generated by trophoblast cell fusion at the maternal-fetal interface. We found that syncytin and type I viral envelope proteins shared similar structural profiling, especially in the regions of N- and C-terminal heptad repeats (NHR and CHR). We expressed the predicted regions of NHR (41 aa) and CHR (34 aa) in syncytin as a native single chain (named 2-helix protein) to characterize it. 2-helix protein exists as a trimer and is highly α-helix, thermo-stable, and denatured by low pH. NHR and CHR could form a protease-resistant complex. The complex structure built by the molecular docking demonstrated that NHR and CHR associated in an antiparallel manner. Overall, the 2-helix protein could form a thermo-stable coiled coil trimer. The fusion core structure of syncytin was first demonstrated in endogenous retrovirus. These results support the explanation how syncytin mediates cytotrophoblast cell fusion involved in placental morphogenesis

  5. A novel fusion protein of IP10-scFv retains antibody specificity and chemokine function

    Energy Technology Data Exchange (ETDEWEB)

    Junqing, Guo; Liu, Chen; Hongwu, Ai; Jiannian, Jing; Jiyong, Zhou; Chuyu, Zhang; Shangyou, You

    2004-07-23

    We combined the specificity of tumor-specific antibody with the chemokine function of interferon-{gamma} inducible protein 10 (IP-10) to recruit immune effector cells in the vicinity of tumor cells. A novel fusion protein of IP10-scFv was constructed by fusing mouse IP-10 to V{sub H} region of single-chain Fv fragment (scFv) against acidic isoferritin (AIF), and expressed in NS0 murine myeloma cells. The IP10-scFv fusion protein was shown to maintain the specificity of the antiAIF scFv with similar affinity constant, and bind to the human hepatocarcinoma SMMC 7721 cells secreting AIF as well as the activated mouse T lymphocytes expressing CXCR3 receptor. Furthermore, the IP10-scFv protein either in solution or bound on the surface of SMMC 7721 cells induced significant chemotaxis of mouse T cells in vitro. The results indicate that the IP10-scFv fusion protein possesses both bioactivities of the tumor-specific antibody and IP-10 chemokine, suggesting its possibility to induce an enhanced immune response against the residual tumor cells in vivo.

  6. A novel fusion protein of IP10-scFv retains antibody specificity and chemokine function

    International Nuclear Information System (INIS)

    Guo Junqing; Chen Liu; Ai Hongwu; Jing Jiannian; Zhou Jiyong; Zhang Chuyu; You Shangyou

    2004-01-01

    We combined the specificity of tumor-specific antibody with the chemokine function of interferon-γ inducible protein 10 (IP-10) to recruit immune effector cells in the vicinity of tumor cells. A novel fusion protein of IP10-scFv was constructed by fusing mouse IP-10 to V H region of single-chain Fv fragment (scFv) against acidic isoferritin (AIF), and expressed in NS0 murine myeloma cells. The IP10-scFv fusion protein was shown to maintain the specificity of the antiAIF scFv with similar affinity constant, and bind to the human hepatocarcinoma SMMC 7721 cells secreting AIF as well as the activated mouse T lymphocytes expressing CXCR3 receptor. Furthermore, the IP10-scFv protein either in solution or bound on the surface of SMMC 7721 cells induced significant chemotaxis of mouse T cells in vitro. The results indicate that the IP10-scFv fusion protein possesses both bioactivities of the tumor-specific antibody and IP-10 chemokine, suggesting its possibility to induce an enhanced immune response against the residual tumor cells in vivo

  7. Combining random gene fission and rational gene fusion to discover near-infrared fluorescent protein fragments that report on protein-protein interactions.

    Science.gov (United States)

    Pandey, Naresh; Nobles, Christopher L; Zechiedrich, Lynn; Maresso, Anthony W; Silberg, Jonathan J

    2015-05-15

    Gene fission can convert monomeric proteins into two-piece catalysts, reporters, and transcription factors for systems and synthetic biology. However, some proteins can be challenging to fragment without disrupting function, such as near-infrared fluorescent protein (IFP). We describe a directed evolution strategy that can overcome this challenge by randomly fragmenting proteins and concomitantly fusing the protein fragments to pairs of proteins or peptides that associate. We used this method to create libraries that express fragmented IFP as fusions to a pair of associating peptides (IAAL-E3 and IAAL-K3) and proteins (CheA and CheY) and screened for fragmented IFP with detectable near-infrared fluorescence. Thirteen novel fragmented IFPs were identified, all of which arose from backbone fission proximal to the interdomain linker. Either the IAAL-E3 and IAAL-K3 peptides or CheA and CheY proteins could assist with IFP fragment complementation, although the IAAL-E3 and IAAL-K3 peptides consistently yielded higher fluorescence. These results demonstrate how random gene fission can be coupled to rational gene fusion to create libraries enriched in fragmented proteins with AND gate logic that is dependent upon a protein-protein interaction, and they suggest that these near-infrared fluorescent protein fragments will be suitable as reporters for pairs of promoters and protein-protein interactions within whole animals.

  8. The conserved glycine residues in the transmembrane domain of the Semliki Forest virus fusion protein are not required for assembly and fusion

    International Nuclear Information System (INIS)

    Liao Maofu; Kielian, Margaret

    2005-01-01

    The alphavirus Semliki Forest virus (SFV) infects cells via a low pH-triggered fusion reaction mediated by the viral E1 protein. Both the E1 fusion peptide and transmembrane (TM) domain are essential for membrane fusion, but the functional requirements for the TM domain are poorly understood. Here we explored the role of the five TM domain glycine residues, including the highly conserved glycine pair at E1 residues 415/416. SFV mutants with alanine substitutions for individual or all five glycine residues (5G/A) showed growth kinetics and fusion pH dependence similar to those of wild-type SFV. Mutants with increasing substitution of glycine residues showed an increasingly more stringent requirement for cholesterol during fusion. The 5G/A mutant showed decreased fusion kinetics and extent in fluorescent lipid mixing assays. TM domain glycine residues thus are not required for efficient SFV fusion or assembly but can cause subtle effects on the properties of membrane fusion

  9. A Type-2 fuzzy data fusion approach for building reliable weighted protein interaction networks with application in protein complex detection.

    Science.gov (United States)

    Mehranfar, Adele; Ghadiri, Nasser; Kouhsar, Morteza; Golshani, Ashkan

    2017-09-01

    Detecting the protein complexes is an important task in analyzing the protein interaction networks. Although many algorithms predict protein complexes in different ways, surveys on the interaction networks indicate that about 50% of detected interactions are false positives. Consequently, the accuracy of existing methods needs to be improved. In this paper we propose a novel algorithm to detect the protein complexes in 'noisy' protein interaction data. First, we integrate several biological data sources to determine the reliability of each interaction and determine more accurate weights for the interactions. A data fusion component is used for this step, based on the interval type-2 fuzzy voter that provides an efficient combination of the information sources. This fusion component detects the errors and diminishes their effect on the detection protein complexes. So in the first step, the reliability scores have been assigned for every interaction in the network. In the second step, we have proposed a general protein complex detection algorithm by exploiting and adopting the strong points of other algorithms and existing hypotheses regarding real complexes. Finally, the proposed method has been applied for the yeast interaction datasets for predicting the interactions. The results show that our framework has a better performance regarding precision and F-measure than the existing approaches. Copyright © 2017 Elsevier Ltd. All rights reserved.

  10. First generation of fusion power plants: Design and technology. Proceedings of the 2. IAEA technical meeting

    International Nuclear Information System (INIS)

    2008-01-01

    This series of meetings has been initiated under recommendation of the International Fusion Research Council for the IAEA and is expected to initiate, develop and mature ideas on fusion strategy that would be of benefit for all players. The present objectives of this meeting are to provide a forum to discuss concepts, technology and environmental aspects of future fusion power plants, the next step following ITER, their role in future energy mix and to assess a selection of urgent topics aiming at identifying the physics and the technological requirements that ITER and a fusion grade materials developing programme will have to address to support the construction of a DEMO(s) fusion power plant(s) prototype demonstrating viable economics. The meeting was organized in five sessions addressing five topics: - (PPCA) Power Plant Concepts and systems Analysis. - (MCP) Materials analysis/Components design/Plasma requirements - (NE) Non-Electric applications of fusion - (SESE) Social, Economic, Safety and Environmental aspects of fusion - (EP) Energy Policy, strategy and scenario for fusion development. A summary session took place at the end of the meeting. Thirty-three participants representing 12 Countries and 3 International Organizations were present at the meeting

  11. Production of FMDV virus-like particles by a SUMO fusion protein approach in Escherichia coli

    Directory of Open Access Journals (Sweden)

    Liang Shu-Mei

    2009-08-01

    Full Text Available Abstract Virus-like particles (VLPs are formed by the self-assembly of envelope and/or capsid proteins from many viruses. Some VLPs have been proven successful as vaccines, and others have recently found applications as carriers for foreign antigens or as scaffolds in nanoparticle biotechnology. However, production of VLP was usually impeded due to low water-solubility of recombinant virus capsid proteins. Previous studies revealed that virus capsid and envelope proteins were often posttranslationally modified by SUMO in vivo, leading into a hypothesis that SUMO modification might be a common mechanism for virus proteins to retain water-solubility or prevent improper self-aggregation before virus assembly. We then propose a simple approach to produce VLPs of viruses, e.g., foot-and-mouth disease virus (FMDV. An improved SUMO fusion protein system we developed recently was applied to the simultaneous expression of three capsid proteins of FMDV in E. coli. The three SUMO fusion proteins formed a stable heterotrimeric complex. Proteolytic removal of SUMO moieties from the ternary complexes resulted in VLPs with size and shape resembling the authentic FMDV. The method described here can also apply to produce capsid/envelope protein complexes or VLPs of other disease-causing viruses.

  12. Identification of two Th1 cell epitopes on the Babesia bovis-encoded 77-kilodalton merozoite protein (Bb-1) by use of truncated recombinant fusion proteins.

    Science.gov (United States)

    Brown, W C; Zhao, S; Woods, V M; Tripp, C A; Tetzlaff, C L; Heussler, V T; Dobbelaere, D A; Rice-Ficht, A C

    1993-01-01

    Previous studies have demonstrated the serologic and T-cell immunogenicity for cattle of a recombinant form of the apical complex-associated 77-kDa merozite protein of Babesia bovis, designated Bb-1. The present study characterizes the immunogenic epitopes of the Bb-1 protein. A series of recombinant truncated fusion proteins spanning the majority of the Bb-1 protein were expressed in Escherichia coli, and their reactivities with bovine peripheral blood mononuclear cells and T-cell clones derived from B. bovis-immune cattle and with rabbit antibodies were determined. Lymphocytes from two immune cattle were preferentially stimulated by the N-terminal half of the Bb-1 protein (amino acids 23 to 266, termed Bb-1A), localizing the T-cell epitopes to the Bb-1A portion of the molecule. CD4+ T-cell clones derived by stimulation with the intact Bb-1 fusion protein were used to identify two T-cell epitopes in the Bb-1A protein, consisting of amino acids SVVLLSAFSGN VWANEAEVSQVVK and FSDVDKTKSTEKT (residues 23 to 46 and 82 to 94). In contrast, rabbit antiserum raised against the intact fusion protein reacted only with the C-terminal half of the protein (amino acids 267 to 499, termed Bb-1B), which contained 28 tandem repeats of the tetrapeptide PAEK or PAET. Biological assays and Northern (RNA) blot analyses for cytokines revealed that following activation with concanavalin A, T-cell clones reactive against the two Bb-1A epitopes produced interleukin-2, gamma interferon, and tumor necrosis factors beta and alpha, but not interleukin-4, suggesting that the Bb-1 antigen preferentially stimulates the Th1 subset of CD4+ T cells in cattle. The studies described here report for the first time the characterization, by cytokine production, of the Th1 subset of bovine T cells and show that, as in mice, protozoal antigens can induce Th1 cells in ruminants. This first demonstration of B. bovis-encoded Th1 cell epitopes provides a rationale for incorporation of all or part of the Bb-1

  13. US-DOE Fusion-Breeder Program: blanket design and system performance

    International Nuclear Information System (INIS)

    Lee, J.D.

    1983-01-01

    Conceptual design studies are being used to assess the technical and economic feasibility of fusion's potential to produce fissile fuel. A reference design of a fission-suppressed blanket using conventional materials is under development. Theoretically, a fusion breeder that incorporates this fusion-suppressed blanket surrounding a 3000-MW tandem mirror fusion core produces its own tritium plus 5600 kg of 233 U per year. The 233 U could then provide fissile makeup for 21 GWe of light-water reactor (LWR) power using a denatured thorium fuel cycle with full recycle. This is 16 times the net electric power produced by the fusion breeder (1.3 GWe). The cost of electricity from this fusion-fission system is estimated to be only 23% higher than the cost from LWRs that have makeup from U 3 O 8 at present costs (55 $/kg). Nuclear performance, magnetohydrodynamics (MHD), radiation effects, and other issues concerning the fission-suppressed blanket are summarized, as are some of the present and future objectives of the fusion breeder program

  14. Higher accumulation of F1-V fusion recombinant protein in plants after induction of protein body formation.

    Science.gov (United States)

    Alvarez, M Lucrecia; Topal, Emel; Martin, Federico; Cardineau, Guy A

    2010-01-01

    Improving foreign protein accumulation is crucial for enhancing the commercial success of plant-based production systems since product yields have a major influence on process economics. Cereal grain evolved to store large amounts of proteins in tightly organized aggregates. In maize, gamma-Zein is the major storage protein synthesized by the rough endoplasmic reticulum (ER) and stored in specialized organelles called protein bodies (PB). Zera (gamma-Zein ER-accumulating domain) is the N-terminal proline-rich domain of gamma-zein that is sufficient to induce the assembly of PB formation. Fusion of the Zera domain to proteins of interest results in assembly of dense PB-like, ER-derived organelles, containing high concentration of recombinant protein. Our main goal was to increase recombinant protein accumulation in plants in order to enhance the efficiency of orally-delivered plant-made vaccines. It is well known that oral vaccination requires substantially higher doses than parental formulations. As a part of a project to develop a plant-made plague vaccine, we expressed our model antigen, the Yersinia pestis F1-V antigen fusion protein, with and without a fused Zera domain. We demonstrated that Zera-F1-V protein accumulation was at least 3x higher than F1-V alone when expressed in three different host plant systems: Ncotiana benthamiana, Medicago sativa (alfalfa) and Nicotiana tabacum NT1 cells. We confirmed the feasibility of using Zera technology to induce protein body formation in non-seed tissues. Zera expression and accumulation did not affect plant development and growth. These results confirmed the potential exploitation of Zera technology to substantially increase the accumulation of value-added proteins in plants.

  15. G protein betagamma-subunits activated by serotonin mediate presynaptic inhibition by regulating vesicle fusion properties.

    Science.gov (United States)

    Photowala, Huzefa; Blackmer, Trillium; Schwartz, Eric; Hamm, Heidi E; Alford, Simon

    2006-03-14

    Neurotransmitters are thought to be released as quanta, where synaptic vesicles deliver packets of neurotransmitter to the synaptic cleft by fusion with the plasma membrane. However, synaptic vesicles may undergo incomplete fusion. We provide evidence that G protein-coupled receptors inhibit release by causing such incomplete fusion. 5-hydroxytryptamine (5-HT) receptor signaling potently inhibits excitatory postsynaptic currents (EPSCs) between lamprey reticulospinal axons and their postsynaptic targets by a direct action on the vesicle fusion machinery. We show that 5-HT receptor-mediated presynaptic inhibition, at this synapse, involves a reduction in EPSC quantal size. Quantal size was measured directly by comparing unitary quantal amplitudes of paired EPSCs before and during 5-HT application and indirectly by determining the effect of 5-HT on the relationship between mean-evoked EPSC amplitude and variance. Results from FM dye-labeling experiments indicate that 5-HT prevents full fusion of vesicles. 5-HT reduces FM1-43 staining of vesicles with a similar efficacy to its effect on the EPSC. However, destaining of FM1-43-labeled vesicles is abolished by lower concentrations of 5-HT that leave a substantial EPSC. The use of a water-soluble membrane impermeant quenching agent in the extracellular space reduced FM1-43 fluorescence during stimulation in 5-HT. Thus vesicles contact the extracellular space during inhibition of synaptic transmission by 5-HT. We conclude that 5-HT, via free Gbetagamma, prevents the collapse of synaptic vesicles into the presynaptic membrane.

  16. Inhibitory effect of PTD-OD-HA fusion protein on Bcr-Abl in K562 cells

    Directory of Open Access Journals (Sweden)

    Miao GAO

    2012-10-01

    Full Text Available Objective To study the transduction dynamics, location of PTD-OD-HA fusion protein and its interaction with Bcr-Abl oncoprotein in K562 cell lines, and explore the influence of PTD-OD-HA fusion protein on oligomerization and tyrosine kinase activity of Bcr-Abl. Methods PTD-OD-HA fusion protein was labeled with FITC and co-cultured with K562 cells. The transduction efficiency of labeled PTD-OD-HA at different doses and time intervals was observed under fluorescence microscope. The location of labeled PTD-OD-HA fusion protein in K562 cells was detected by confocal microscopy. The interaction of PTD-OD-HA fusion protein with Bcr-Abl oncoprotein was confirmed by coimmunoprecipitation. The phosphorylation of Bcr-Abl oncoprotein was detected by Western blotting. Results PTD-OD-HA fusion protein labeled with FITC was transduced into K562 cells in a dose- and time-dependent manner. PTD-OD-HA fusion protein was located in the cytoplasm of K562 cells and was consistent with the location of Bcr-Abl oncoprotein. The interaction of PTD-OD-HA fusion protein with Bcr-Abl oncoprotein was proved in K562 cells. This interaction could interrupt the homologous oligomerization of Bcr-Abl oncoprotein and reduce the phosphorylation of Bcr-Abl oncoprotein. Conclusion PTD-OD-HA fusion protein could be transduced into K562 cells efficiently, inhibit the oligomerization and reduce the phosphorylation of Bcr-Abl oncoprotein.

  17. IFMIF : International Fusion Materials Irradiation Facility Conceptual Design Activity: Executive summary

    International Nuclear Information System (INIS)

    1997-01-01

    This report is a summary of the results of the Conceptual Design Activity (CDA) on the International Fusion Materials Irradiation Facility (IFMIF), conducted during 1995 and 1996. The activity is under the auspices of the International Energy Agency (IEA) Implementing Agreement for a Programme of Research and Development on Fusion Materials. An IEA Fusion Materials Executive Subcommittee was charged with overseeing the IFMIF-CDA work. Participants in the CDA are the European Union, Japan, and the United States, with the Russian Federation as an associate member

  18. IFMIF : International Fusion Materials Irradiation Facility Conceptual Design Activity: Executive summary

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1997-01-01

    This report is a summary of the results of the Conceptual Design Activity (CDA) on the International Fusion Materials Irradiation Facility (IFMIF), conducted during 1995 and 1996. The activity is under the auspices of the International Energy Agency (IEA) Implementing Agreement for a Programme of Research and Development on Fusion Materials. An IEA Fusion Materials Executive Subcommittee was charged with overseeing the IFMIF-CDA work. Participants in the CDA are the European Union, Japan, and the United States, with the Russian Federation as an associate member.

  19. Design aspects of a multipurpose fusion power plant for desalination and agrochemical processes

    International Nuclear Information System (INIS)

    Sabri, Z.A.

    1975-02-01

    A description is given of the skeletal structure of a multipurpose fusion power plant, designed for desalination and agrochemicals production. The plant is a complex that comprises dual purpose power and desalination units, separation and processing units for recovery of soluble salts in the effluent of the desalination unit, mariculture units for production of algae for food and as food for shrimp and other fish species. The electrical power unit is a two-component fusion device that burns deuterium and helium-3 utilizing a fast fusion cycle

  20. The reversed-field-pinch (RFP) fusion neutron source: A conceptual design

    International Nuclear Information System (INIS)

    Bathke, C.G.; Krakowski, R.A.; Miller, R.L.; Werley, K.A.

    1989-01-01

    The conceptual design of an ohmically heated, reversed-field pinch (RFP) operating at ∼5-MW/m 2 steady-state DT fusion neutron wall loading and ∼124-MW total fusion power is presented. These results are useful in projecting the development of a cost effective, low input power (∼206 MW) source of DT neutrons for large-volume (∼10 m 3 ), high-fluence (3.4 MW yr/m 2 ) fusion nuclear materials and technology testing. 19 refs., 15 figs., 9 tabs

  1. IFMIF : International Fusion Materials Irradiation Facility Conceptual Design Activity: Final report

    International Nuclear Information System (INIS)

    Martone, M.

    1997-01-01

    This report documents the results of the Conceptual Design Activity (CDA) on the International Fusion Materials Irradiation Facility (IFMIF), conducted during 1995 and 1996. The activity is under the auspices of the International Energy Agency (IEA) Implementing Agreement for a Programme of Research and Development on Fusion Materials. An IEA Fusion Materials Executive Subcommittee was charged with overseeing the IFMIF-CDA work. Participants in the CDA are the European Union, Japan, and the United States, with the Russian Federation as an associate member

  2. IFMIF : International Fusion Materials Irradiation Facility Conceptual Design Activity: Final report

    Energy Technology Data Exchange (ETDEWEB)

    Martone, M [ENEA, Centro Ricerche Frascati, Rome (Italy)

    1997-01-01

    This report documents the results of the Conceptual Design Activity (CDA) on the International Fusion Materials Irradiation Facility (IFMIF), conducted during 1995 and 1996. The activity is under the auspices of the International Energy Agency (IEA) Implementing Agreement for a Programme of Research and Development on Fusion Materials. An IEA Fusion Materials Executive Subcommittee was charged with overseeing the IFMIF-CDA work. Participants in the CDA are the European Union, Japan, and the United States, with the Russian Federation as an associate member.

  3. Progress on the conceptual design of a mirror hybrid fusion--fission reactor

    International Nuclear Information System (INIS)

    Moir, R.W.; Lee, J.D.; Burleigh, R.J.

    1975-01-01

    A conceptual design study was made of a fusion-fission reactor for the purpose of producing fissile material and electricity. The fusion component is a D-T plasma confined by a pair of magnetic mirror coils in a Yin-Yang configuration and is sustained by neutral beam injection. The neutrons from the fusion plasma drive the fission assembly which is composed of natural uranium carbide fuel rods clad with stainless steel and helium cooled. It was shown conceptually how the reactor might be built using essentially present-day technology and how the uranium-bearing blanket modules can be routinely changed to allow separation of the bred fissile fuel

  4. Analysis of nuclear export using photoactivatable GFP fusion proteins and interspecies heterokaryons.

    Science.gov (United States)

    Nakrieko, Kerry-Ann; Ivanova, Iordanka A; Dagnino, Lina

    2010-01-01

    In this chapter, we review protocols for the analysis of nucleocytoplasmic shuttling of transcription factors and nuclear proteins, using two different approaches. The first involves the use of photoactivatable forms of the protein of interest by fusion to photoactivatable green fluorescent protein to follow its movement out of the nucleus by live-cell confocal microscopy. This methodology allows for the kinetic characterization of protein movements as well as measurement of steady-state levels. In a second procedure to assess the ability of a nuclear protein to move into and out of the nucleus, we describe the use of interspecies heterokaryon assays, which provide a measurement of steady-state distribution. These technologies are directly applicable to the analysis of nucleocytoplasmic movements not only of transcription factors, but also other nuclear proteins.

  5. Design system for in-vessel mainipulator of fusion reactor 'DESIM'

    International Nuclear Information System (INIS)

    Adachi, Junihci; Kobayashi, Takeshi; Ise, Hideo; Sato, Keisuke; Matsuda, Hirotsugu

    1989-01-01

    A computer aided design system 'DESIM' for the in-vessel manipulators of nuclear fusion reactors has been developed to design the manipulators efficiently. The DESIM consists of the following subsystems: (1) the design system for arm mechanisms to realize optimum manipulation performance in the specified workspace; (2) the robot simulator to study manipulator movement, postures and interference problems; (3) the CAD system which is used to define the structure object data for robots, and the interface system for the data conversion from the CAD system to the robot simulator. The DESIM has been used to design the in-vessel manipulator for the Fusion Experimental Reactor (FER) to confirm the effectiveness. (author)

  6. Conceptual design of a fission-based integrated test facility for fusion reactor components

    International Nuclear Information System (INIS)

    Watts, K.D.; Deis, G.A.; Hsu, P.Y.S.; Longhurst, G.R.; Masson, L.S.; Miller, L.G.

    1982-01-01

    The testing of fusion materials and components in fission reactors will become increasingly important because of lack of fusion engineering test devices in the immediate future and the increasing long-term demand for fusion testing when a fusion reactor test station becomes available. This paper presents the conceptual design of a fission-based Integrated Test Facility (ITF) developed by EG and G Idaho. This facility can accommodate entire first wall/blanket (FW/B) test modules such as those proposed for INTOR and can also accommodate smaller cylindrical modules similar to those designed by Oak Ridge National laboratory (ORNL) and Westinghouse. In addition, the facility can be used to test bulk breeder blanket materials, materials for tritium permeation, and components for performance in a nuclear environment. The ITF provides a cyclic neutron/gamma flux as well as the numerous module and experiment support functions required for truly integrated tests

  7. Purification method for recombinant proteins based on a fusion between the target protein and the C-terminus of calmodulin

    Science.gov (United States)

    Schauer-Vukasinovic, Vesna; Deo, Sapna K.; Daunert, Sylvia

    2002-01-01

    Calmodulin (CaM) was used as an affinity tail to facilitate the purification of the green fluorescent protein (GFP), which was used as a model target protein. The protein GFP was fused to the C-terminus of CaM, and a factor Xa cleavage site was introduced between the two proteins. A CaM-GFP fusion protein was expressed in E. coli and purified on a phenothiazine-derivatized silica column. CaM binds to the phenothiazine on the column in a Ca(2+)-dependent fashion and it was, therefore, used as an affinity tail for the purification of GFP. The fusion protein bound to the affinity column was then subjected to a proteolytic digestion with factor Xa. Pure GFP was eluted with a Ca(2+)-containing buffer, while CaM was eluted later with a buffer containing the Ca(2+)-chelating agent EGTA. The purity of the isolated GFP was verified by SDS-PAGE, and the fluorescence properties of the purified GFP were characterized.

  8. The B isozyme creatine kinase is active as a fusion protein in Escherichia coli

    International Nuclear Information System (INIS)

    Koretsky, A.P.; Traxler, B.A.

    1989-01-01

    A cDNA encoding the B isozyme of creatine kinase CK B has been expressed in Escherichia coli from a fusion with lacZ carried by λgtll. Western blots indicate that a stable polypeptide with the appropriate mobility for the Β-galactosidase-creatine kinase Β-gal-CK B ) fusion protein cross-reacts with both Β-gal and CK B antiserum. No significant CK activity is detected in control E. coli; however, extracts from cells containing the λgtll-CK B construct have a CK activity of 1.54j0.07 μmol/min per mg protein. The fusion protein appears to provide this activity bacause immunoprecipitation of protein with Β-gal antiserum leads to a loss of CK activity from extracts. That the enzyme is active in vivo was demonstrated by detection of a phosphocreatine (PCr) peak in the 31 P NMR spectrum from E. coli grown on medium supplemented with creatine. As in mammalian brain and muscle, the PCr peak detected was sensitive to the energy status of the E. coli. (author). 17 refs.; 3 figs.; 1 tab

  9. Primary heat transfer loop design for the Cascade inertial confinement fusion reactor

    International Nuclear Information System (INIS)

    Murray, K.A.; McDowell, M.W.

    1984-05-01

    This study investigates a heat exchanger and balance of plant design to accompany the Cascade inertial confinement fusion reaction chamber concept. The concept uses solid Li 2 O or other lithium-ceramic granules, held to the wall of a rotating reaction chamber by centrifugal action, as a tritium breeding blanket and first wall protection. The Li 2 O granules enter the chamber at 800 K and exit at 1200 K after absorbing the thermal energy produced by the fusion process

  10. Bulk-shield design for the Fusion Materials Irradiation Test facility

    International Nuclear Information System (INIS)

    Carter, L.L.; Mann, F.M.; Morford, R.J.; Johnson, D.L.; Huang, S.T.

    1982-07-01

    The accelerator-based Fusion Materials Irradiation Test (FMIT) facility will provide a high-fluence, fusion-like radiation environment for the testing of materials. While the neutron spectrum produced in the forward direction by the 35 MeV deuterons incident upon a flowing lithium target is characterized by a broad peak around 14 MeV, a high energy tail extends up to about 50 MeV. Some shield design considerations are reviewed

  11. Targeted recombinant fusion proteins of IFNγ and mimetic IFNγ with PDGFβR bicyclic peptide inhibits liver fibrogenesis in vivo.

    Directory of Open Access Journals (Sweden)

    Ruchi Bansal

    Full Text Available Hepatic stellate cells (HSCs, following transdifferentiation to myofibroblasts plays a key role in liver fibrosis. Therefore, attempts to attenuate this myofibroblastic phenotype would be a promising therapeutic approach. Interferon gamma (IFNγ is a potent anti-fibrotic cytokine, but its pleiotropic receptor expression leading to severe adverse effects has limited its clinical application. Since, activated HSC express high-level of platelet derived growth factor beta receptor (PDGFβR, we investigated the potential of PDGFβR-specific targeting of IFNγ and its signaling peptide that lacks IFNγR binding site (mimetic IFNγ or mimIFNγ in liver fibrosis. We prepared DNA constructs expressing IFNγ, mimIFNγ or BiPPB (PDGFβR-specific bicyclic peptide-IFNγ, BiPPB-mimIFNγ fusion proteins. Both chimeric proteins alongwith IFNγ and mimIFNγ were produced in E.coli. The expressed proteins were purified and analyzed for PDGFβR-specific binding and in vitro effects. Subsequently, these recombinant proteins were investigated for the liver uptake (pSTAT1α signaling pathway, for anti-fibrotic effects and adverse effects (platelet counts in CCl4-induced liver fibrogenesis in mice. The purified HSC-targeted IFNγ and mimIFNγ fusion proteins showed PDGFβR-specific binding and significantly reduced TGFβ-induced collagen-I expression in human HSC (LX2 cells, while mouse IFNγ and mimIFNγ did not show any effect. Conversely, mouse IFNγ and BiPPB-IFNγ induced activation and dose-dependent nitric oxide release in mouse macrophages (express IFNγR while lack PDGFβR, which was not observed with mimIFNγ and BiPPB-mimIFNγ, due to the lack of IFNγR binding sites. In vivo, targeted BiPPB-IFNγ and BiPPB-mimIFNγ significantly activated intrahepatic IFNγ-signaling pathway compared to IFNγ and mimIFNγ suggesting increased liver accumulation. Furthermore, the targeted fusion proteins ameliorated liver fibrogenesis in mice by significantly reducing

  12. Blood-brain barrier drug delivery of IgG fusion proteins with a transferrin receptor monoclonal antibody.

    Science.gov (United States)

    Pardridge, William M

    2015-02-01

    Biologic drugs are large molecules that do not cross the blood- brain barrier (BBB). Brain penetration is possible following the re-engineering of the biologic drug as an IgG fusion protein. The IgG domain is a MAb against an endogenous BBB receptor such as the transferrin receptor (TfR). The TfRMAb acts as a molecular Trojan horse to ferry the fused biologic drug into the brain via receptor-mediated transport on the endogenous BBB TfR. This review discusses TfR isoforms, models of BBB transport of transferrin and TfRMAbs, and the genetic engineering of TfRMAb fusion proteins, including BBB penetrating IgG-neurotrophins, IgG-decoy receptors, IgG-lysosomal enzyme therapeutics and IgG-avidin fusion proteins, as well as BBB transport of bispecific antibodies formed by fusion of a therapeutic antibody to a TfRMAb targeting antibody. Also discussed are quantitative aspects of the plasma pharmacokinetics and brain uptake of TfRMAb fusion proteins, as compared to the brain uptake of small molecules, and therapeutic applications of TfRMAb fusion proteins in mouse models of neural disease, including Parkinson's disease, stroke, Alzheimer's disease and lysosomal storage disorders. The review covers the engineering of TfRMAb-avidin fusion proteins for BBB targeted delivery of biotinylated peptide radiopharmaceuticals, low-affinity TfRMAb Trojan horses and the safety pharmacology of chronic administration of TfRMAb fusion proteins. The BBB delivery of biologic drugs is possible following re-engineering as a fusion protein with a molecular Trojan horse such as a TfRMAb. The efficacy of this technology will be determined by the outcome of future clinical trials.

  13. International Fusion Materials Irradiation Facility conceptual design activity. Present status and perspective

    International Nuclear Information System (INIS)

    Kondo, Tatsuo; Noda, Kenji; Oyama, Yukio

    1998-01-01

    For developing the materials for nuclear fusion reactors, it is indispensable to study on the neutron irradiation behavior under fusion reactor conditions, but there is not any high energy neutron irradiation facility that can simulate fusion reactor conditions at present. Therefore, the investigation of the IFMIF was begun jointly by Japan, USA, Europe and Russia following the initiative of IEA. The conceptual design activities were completed in 1997. As to the background and the course, the present status of the research on heavy irradiation and the testing means for fusion materials, the requirement and the technical basis of high energy neutron irradiation, and the international joint design activities are reported. The materials for fusion reactors are exposed to the neutron irradiation with the energy spectra up to 14 MeV. The requirements from the users that the IFMIF should satisfy, the demand of the tests for the materials of prototype and demonstration fusion reactors and the evaluation of the neutron field characteristics of the IFMIF are discussed. As to the conceptual design of the IFMIF, the whole constitution, the operational mode, accelerator system and target system are described. (K.I.)

  14. Solid-State Nuclear Magnetic Resonance Investigation of the Structural Topology and Lipid Interactions of a Viral Fusion Protein Chimera Containing the Fusion Peptide and Transmembrane Domain.

    Science.gov (United States)

    Yao, Hongwei; Lee, Myungwoon; Liao, Shu-Yu; Hong, Mei

    2016-12-13

    The fusion peptide (FP) and transmembrane domain (TMD) of viral fusion proteins play important roles during virus-cell membrane fusion, by inducing membrane curvature and transient dehydration. The structure of the water-soluble ectodomain of viral fusion proteins has been extensively studied crystallographically, but the structures of the FP and TMD bound to phospholipid membranes are not well understood. We recently investigated the conformations and lipid interactions of the separate FP and TMD peptides of parainfluenza virus 5 (PIV5) fusion protein F using solid-state nuclear magnetic resonance. These studies provide structural information about the two domains when they are spatially well separated in the fusion process. To investigate how these two domains are structured relative to each other in the postfusion state, when the ectodomain forms a six-helix bundle that is thought to force the FP and TMD together in the membrane, we have now expressed and purified a chimera of the FP and TMD, connected by a Gly-Lys linker, and measured the chemical shifts and interdomain contacts of the protein in several lipid membranes. The FP-TMD chimera exhibits α-helical chemical shifts in all the membranes examined and does not cause strong curvature of lamellar membranes or membranes with negative spontaneous curvature. These properties differ qualitatively from those of the separate peptides, indicating that the FP and TMD interact with each other in the lipid membrane. However, no 13 C- 13 C cross peaks are observed in two-dimensional correlation spectra, suggesting that the two helices are not tightly associated. These results suggest that the ectodomain six-helix bundle does not propagate into the membrane to the two hydrophobic termini. However, the loosely associated FP and TMD helices are found to generate significant negative Gaussian curvature to membranes that possess spontaneous positive curvature, consistent with the notion that the FP-TMD assembly may

  15. Nuclear localization and transactivating capacities of the papillary renal cell carcinoma-associated TFE3 and PRCC (fusion) proteins

    NARCIS (Netherlands)

    Weterman, M. A. J.; van Groningen, J. J.; Jansen, A.; van Kessel, A. G.

    2000-01-01

    The papillary renal cell carcinoma-associated t(X;1)(p11;q21) leads to fusion of the transcription factor TFE3 gene on the X-chromosome to a novel gene, PRCC, on chromosome 1. As a result, two putative fusion proteins are formed: PRCCTFE3, which contains all known domains for DNA binding,

  16. Functional NifD-K fusion protein in Azotobacter vinelandii is a homodimeric complex equivalent to the native heterotetrameric MoFe protein

    International Nuclear Information System (INIS)

    Lahiri, Surobhi; Pulakat, Lakshmi; Gavini, Nara

    2005-01-01

    The MoFe protein of the complex metalloenzyme nitrogenase folds as a heterotetramer containing two copies each of the homologous α and β subunits, encoded by the nifD and the nifK genes respectively. Recently, the functional expression of a fusion NifD-K protein of nitrogenase was demonstrated in Azotobacter vinelandii, strongly implying that the MoFe protein is flexible as it could accommodate major structural changes, yet remain functional [M.H. Suh, L. Pulakat, N. Gavini, J. Biol. Chem. 278 (2003) 5353-5360]. This finding led us to further explore the type of interaction between the fused MoFe protein units. We aimed to determine whether an interaction exists between the two fusion MoFe proteins to form a homodimer that is equivalent to native heterotetrameric MoFe protein. Using the Bacteriomatch Two-Hybrid System, translationally fused constructs of NifD-K (fusion) with the full-length λCI of the pBT bait vector and also NifD-K (fusion) with the N-terminal α-RNAP of the pTRG target vector were made. To compare the extent of interaction between the fused NifD-K proteins to that of the β-β interactions in the native MoFe protein, we proceeded to generate translationally fused constructs of NifK with the α-RNAP of the pTRG vector and λCI protein of the pBT vector. The strength of the interaction between the proteins in study was determined by measuring the β-galactosidase activity and extent of ampicillin resistance of the colonies expressing these proteins. This analysis demonstrated that direct protein-protein interaction exists between NifD-K fusion proteins, suggesting that they exist as homodimers. As the interaction takes place at the β-interfaces of the NifD-K fusion proteins, we propose that these homodimers of NifD-K fusion protein may function in a similar manner as that of the heterotetrameric native MoFe protein. The observation that the extent of protein-protein interaction between the β-subunits of the native MoFe protein in Bacterio

  17. Determination of the topology of endoplasmic reticulum membrane proteins using redox-sensitive green-fluorescence protein fusions.

    Science.gov (United States)

    Tsachaki, Maria; Birk, Julia; Egert, Aurélie; Odermatt, Alex

    2015-07-01

    Membrane proteins of the endoplasmic reticulum (ER) are involved in a wide array of essential cellular functions. Identification of the topology of membrane proteins can provide significant insight into their mechanisms of action and biological roles. This is particularly important for membrane enzymes, since their topology determines the subcellular site where a biochemical reaction takes place and the dependence on luminal or cytosolic co-factor pools and substrates. The methods currently available for the determination of topology of proteins are rather laborious and require post-lysis or post-fixation manipulation of cells. In this work, we have developed a simple method for defining intracellular localization and topology of ER membrane proteins in living cells, based on the fusion of the respective protein with redox-sensitive green-fluorescent protein (roGFP). We validated the method and demonstrated that roGFP fusion proteins constitute a reliable tool for the study of ER membrane protein topology, using as control microsomal 11β-hydroxysteroid dehydrogenase (11β-HSD) proteins whose topology has been resolved, and comparing with an independent approach. We then implemented this method to determine the membrane topology of six microsomal members of the 17β-hydroxysteroid dehydrogenase (17β-HSD) family. The results revealed a luminal orientation of the catalytic site for three enzymes, i.e. 17β-HSD6, 7 and 12. Knowledge of the intracellular location of the catalytic site of these enzymes will enable future studies on their biological functions and on the role of the luminal co-factor pool. Copyright © 2015 Elsevier B.V. All rights reserved.

  18. The RCC-MR design code for LMFBR components. A useful basic for fusion reactor design tools development

    International Nuclear Information System (INIS)

    Acker, D.; Chevereau, G.

    1985-11-01

    LMFBR and fusion reactors exhibit common features with regard to structural materials (Stainless steels), temperature service level (550-600 0 C), loading types. So, design and construction rules used in France for LMFBR, that is to say RCC-MR Code, can constitute a good basis for fusion reactors design. Some original aspects of RCC-MR design rules are described, relating to unsignificant creep, ratchetting effect, fatigue and creep damage limits, creep damage evaluation, fatigue damage evaluation, buckling. The main originality of RCC-MR consists to propose comprehensive simplified rules based on elastic calculations and extended from classical cold temperatures to the elevated temperature domain

  19. The RCC-MR design code for LMFBR components. A useful basis for fusion reactor design tools development

    International Nuclear Information System (INIS)

    Acker, D.; Chevereau, G.

    1986-01-01

    LMFBR and fusion reactors exhibit common features with regard to structural materials, temperature service level, loading types. So, design and construction rules used in France for LMFBR, that is to say RCC-MR Code, can constitute a good basis for fusion reactors design. Some original aspects of RCC-MR design rules are described, relating to unsignificant creep, ratchetting effect, fatigue and creep damage limits, creep damage evaluation, fatigue damage evaluation, buckling. The main originality of RCC-MR consists to propose comprehensive simplified rules based on elastic calculations and extended from classical cold temperatures to the elevated temperature domain. (author)

  20. Pioglitazone Induces a Proadipogenic Antitumor Response in Mice with PAX8-PPARγ Fusion Protein Thyroid Carcinoma

    OpenAIRE

    Dobson, Melissa E.; Diallo-Krou, Ericka; Grachtchouk, Vladimir; Yu, Jingcheng; Colby, Lesley A.; Wilkinson, John E.; Giordano, Thomas J.; Koenig, Ronald J.

    2011-01-01

    Approximately 35% of follicular thyroid carcinomas harbor a chromosomal translocation that results in expression of a paired box gene 8-peroxisome proliferator-activated receptor γ gene (PPARγ) fusion protein (PPFP). To better understand the oncogenic role of PPFP and its relationship to endogenous PPARγ, we generated a transgenic mouse model that combines Cre-dependent PPFP expression (PPFP;Cre) with homozygous deletion of floxed Pten (PtenFF;Cre), both thyroid specific. Although neither PPF...

  1. A recombinant fusion protein-based, fluorescent protease assay for high throughput-compatible substrate screening.

    Science.gov (United States)

    Bozóki, Beáta; Gazda, Lívia; Tóth, Ferenc; Miczi, Márió; Mótyán, János András; Tőzsér, József

    2018-01-01

    In connection with the intensive investigation of proteases, several methods have been developed for analysis of the substrate specificity. Due to the great number of proteases and the expected target molecules to be analyzed, time- and cost-efficient high-throughput screening (HTS) methods are preferred. Here we describe the development and application of a separation-based HTS-compatible fluorescent protease assay, which is based on the use of recombinant fusion proteins as substrates of proteases. The protein substrates used in this assay consists of N-terminal (hexahistidine and maltose binding protein) fusion tags, cleavage sequences of the tobacco etch virus (TEV) and HIV-1 proteases, and a C-terminal fluorescent protein (mApple or mTurquoise2). The assay is based on the fluorimetric detection of the fluorescent proteins, which are released from the magnetic bead-attached substrates by the proteolytic cleavage. The protease assay has been applied for activity measurements of TEV and HIV-1 proteases to test the suitability of the system for enzyme kinetic measurements, inhibition studies, and determination of pH optimum. We also found that denatured fluorescent proteins can be renatured after SDS-PAGE of denaturing conditions, but showed differences in their renaturation abilities. After in-gel renaturation both substrates and cleavage products can be identified by in-gel UV detection. Copyright © 2017 Elsevier Inc. All rights reserved.

  2. TALE-PvuII Fusion Proteins – Novel Tools for Gene Targeting

    Science.gov (United States)

    Yanik, Mert; Alzubi, Jamal; Lahaye, Thomas; Cathomen, Toni; Pingoud, Alfred; Wende, Wolfgang

    2013-01-01

    Zinc finger nucleases (ZFNs) consist of zinc fingers as DNA-binding module and the non-specific DNA-cleavage domain of the restriction endonuclease FokI as DNA-cleavage module. This architecture is also used by TALE nucleases (TALENs), in which the DNA-binding modules of the ZFNs have been replaced by DNA-binding domains based on transcription activator like effector (TALE) proteins. Both TALENs and ZFNs are programmable nucleases which rely on the dimerization of FokI to induce double-strand DNA cleavage at the target site after recognition of the target DNA by the respective DNA-binding module. TALENs seem to have an advantage over ZFNs, as the assembly of TALE proteins is easier than that of ZFNs. Here, we present evidence that variant TALENs can be produced by replacing the catalytic domain of FokI with the restriction endonuclease PvuII. These fusion proteins recognize only the composite recognition site consisting of the target site of the TALE protein and the PvuII recognition sequence (addressed site), but not isolated TALE or PvuII recognition sites (unaddressed sites), even at high excess of protein over DNA and long incubation times. In vitro, their preference for an addressed over an unaddressed site is > 34,000-fold. Moreover, TALE-PvuII fusion proteins are active in cellula with minimal cytotoxicity. PMID:24349308

  3. TALE-PvuII fusion proteins--novel tools for gene targeting.

    Science.gov (United States)

    Yanik, Mert; Alzubi, Jamal; Lahaye, Thomas; Cathomen, Toni; Pingoud, Alfred; Wende, Wolfgang

    2013-01-01

    Zinc finger nucleases (ZFNs) consist of zinc fingers as DNA-binding module and the non-specific DNA-cleavage domain of the restriction endonuclease FokI as DNA-cleavage module. This architecture is also used by TALE nucleases (TALENs), in which the DNA-binding modules of the ZFNs have been replaced by DNA-binding domains based on transcription activator like effector (TALE) proteins. Both TALENs and ZFNs are programmable nucleases which rely on the dimerization of FokI to induce double-strand DNA cleavage at the target site after recognition of the target DNA by the respective DNA-binding module. TALENs seem to have an advantage over ZFNs, as the assembly of TALE proteins is easier than that of ZFNs. Here, we present evidence that variant TALENs can be produced by replacing the catalytic domain of FokI with the restriction endonuclease PvuII. These fusion proteins recognize only the composite recognition site consisting of the target site of the TALE protein and the PvuII recognition sequence (addressed site), but not isolated TALE or PvuII recognition sites (unaddressed sites), even at high excess of protein over DNA and long incubation times. In vitro, their preference for an addressed over an unaddressed site is > 34,000-fold. Moreover, TALE-PvuII fusion proteins are active in cellula with minimal cytotoxicity.

  4. IQCJ-SCHIP1, a novel fusion transcript encoding a calmodulin-binding IQ motif protein

    International Nuclear Information System (INIS)

    Kwasnicka-Crawford, Dorota A.; Carson, Andrew R.; Scherer, Stephen W.

    2006-01-01

    The existence of transcripts that span two adjacent, independent genes is considered rare in the human genome. This study characterizes a novel human fusion gene named IQCJ-SCHIP1. IQCJ-SCHIP1 is the longest isoform of a complex transcriptional unit that bridges two separate genes that encode distinct proteins, IQCJ, a novel IQ motif containing protein and SCHIP1, a schwannomin interacting protein that has been previously shown to interact with the Neurofibromatosis type 2 (NF2) protein. IQCJ-SCHIP1 is located on the chromosome 3q25 and comprises a 1692-bp transcript encompassing 11 exons spanning 828 kb of the genomic DNA. We show that IQCJ-SCHIP1 mRNA is highly expressed in the brain. Protein encoded by the IQCJ-SCHIP1 gene was localized to cytoplasm and actin-rich regions and in differentiated PC12 cells was also seen in neurite extensions

  5. Conceptual design study of quasi-steady state fusion experimental reactor (FEQ-Q), part 1

    International Nuclear Information System (INIS)

    1985-12-01

    Since 1980 the design study has been conducted at JAERI for the Fusion Experimental Reactor (FER) which has been proposed to be the next machine to JT-60 in the Japanese long term program of fusion reactor development. Starting from 1984 JER design is being reviewed and redesigned. This report is a part of the interim report which describes the results obtained in the review and redesign activities in FY 1984. The results of the following design items are included; core plasma, reactor structure, reactor core components, magnets. (author)

  6. Conceptual design study of quasi-steady state fusion experimental reactor (FER-Q), part 2

    International Nuclear Information System (INIS)

    1985-12-01

    Since 1980 the design study has been conducted at JAERI for the Fusion Experimental Reactor (FER) which has been proposed to be the next machine to JT-60 in the Japanese long term program of fusion reactor development. Starting from 1984 FER design is being reviewed and redesigned. This report is a part of the interim report which describes the results obtained in the review and redesign activities in FY 1984. The results of the following design items are included: heating/current drive system, plasma position control, power supply, diagnostics, neutronics, blanket test module, repair and maintenance and safety. (author)

  7. Antibody-cytokine fusion proteins for treatment of cancer: engineering cytokines for improved efficacy and safety.

    Science.gov (United States)

    Young, Patricia A; Morrison, Sherie L; Timmerman, John M

    2014-10-01

    The true potential of cytokine therapies in cancer treatment is limited by the inability to deliver optimal concentrations into tumor sites due to dose-limiting systemic toxicities. To maximize the efficacy of cytokine therapy, recombinant antibody-cytokine fusion proteins have been constructed by a number of groups to harness the tumor-targeting ability of monoclonal antibodies. The aim is to guide cytokines specifically to tumor sites where they might stimulate more optimal anti-tumor immune responses while avoiding the systemic toxicities of free cytokine therapy. Antibody-cytokine fusion proteins containing interleukin (IL)-2, IL-12, IL-21, tumor necrosis factor (TNF)α, and interferons (IFNs) α, β, and γ have been constructed and have shown anti-tumor activity in preclinical and early-phase clinical studies. Future priorities for development of this technology include optimization of tumor targeting, bioactivity of the fused cytokine, and choice of appropriate agents for combination therapies. This review is intended to serve as a framework for engineering an ideal antibody-cytokine fusion protein, focusing on previously developed constructs and their clinical trial results. Copyright © 2014 Elsevier Inc. All rights reserved.

  8. Purification, crystallization and X-ray diffraction analysis of human dynamin-related protein 1 GTPase-GED fusion protein

    International Nuclear Information System (INIS)

    Klinglmayr, Eva; Wenger, Julia; Mayr, Sandra; Bossy-Wetzel, Ella; Puehringer, Sandra

    2012-01-01

    The crystallization and initial diffraction analysis of human Drp1 GTPase-GED fusion protein are reported. The mechano-enzyme dynamin-related protein 1 plays an important role in mitochondrial fission and is implicated in cell physiology. Dysregulation of Drp1 is associated with abnormal mitochondrial dynamics and neuronal damage. Drp1 shares structural and functional similarities with dynamin 1 with respect to domain organization, ability to self-assemble into spiral-like oligomers and GTP-cycle-dependent membrane scission. Structural studies of human dynamin-1 have greatly improved the understanding of this prototypical member of the dynamin superfamily. However, high-resolution structural information for full-length human Drp1 covering the GTPase domain, the middle domain and the GTPase effector domain (GED) is still lacking. In order to obtain mechanistic insights into the catalytic activity, a nucleotide-free GTPase-GED fusion protein of human Drp1 was expressed, purified and crystallized. Initial X-ray diffraction experiments yielded data to 2.67 Å resolution. The hexagonal-shaped crystals belonged to space group P2 1 2 1 2, with unit-cell parameters a = 53.59, b = 151.65, c = 43.53 Å, one molecule per asymmetric unit and a solvent content of 42%. Expression of selenomethionine-labelled protein is currently in progress. Here, the expression, purification, crystallization and X-ray diffraction analysis of the Drp1 GTPase-GED fusion protein are presented, which form a basis for more detailed structural and biophysical analysis

  9. Expression and activity analysis of a new fusion protein targeting ovarian cancer cells.

    Science.gov (United States)

    Su, Manman; Chang, Weiqin; Wang, Dingding; Cui, Manhua; Lin, Yang; Wu, Shuying; Xu, Tianmin

    2015-09-01

    The aim of the present study was to develop a new therapeutic drug to improve the prognosis of ovarian cancer patients. Human urokinase-type plasminogen activator (uPA)17-34-kunitz-type protease inhibitor (KPI) eukaryotic expression vector was constructed and recombinant human uPA17-34-KPI (rhuPA17-34-KPI) in P. pastoris was expressed. In the present study, the DNA sequences that encode uPA 17-34 amino acids were created according to the native amino acids sequence and inserted into the KPI-pPICZαC vector, which was constructed. Then, uPA17‑34-KPI-pPICZαC was transformed into P. pastoris X-33, and rhuPA17-34-KPI was expressed by induction of methanol. The bioactivities of a recombinant fusion protein were detected with trypsin inhibition analysis, and the inhibitory effects on the growth of ovarian cancer cells were identified using the TUNEL assay, in vitro wound‑healing assay and Matrigel model analysis. The results of the DNA sequence analysis of the recombinant vector uPA17-34-KPI‑pPICZα demonstrated that the DNA‑encoding human uPA 17-34 amino acids, 285-288 amino acids of amyloid precursor protein (APP) and 1-57 amino acids of KPI were correctly inserted into the pPICZαC vector. Following induction by methonal, the fusion protein with a molecular weight of 8.8 kDa was observed using SDS-PAGE and western blot analysis. RhuPA17-34-KPI was expressed in P. pastoris with a yield of 50 mg/l in a 50-ml tube. The recombinant fusion protein was able to inhibit the activity of trypsin, inhibit growth and induce apoptosis of SKOV3 cells, and inhibit the invasion and metastasis of ovarian cancer cells. By considering uPA17-34 amino acid specific binding uPAR as the targeted part of fusion protein and utilizing the serine protease inhibitor activity of KPI, it was found that the recombinant fusion protein uPA17-34-KPI inhibited the invasion and metastasis of ovarian tumors, and may therefore be regarded as effective in targeted treatment.

  10. Employment of colorimetric enzyme assay for monitoring expression and solubility of GST fusion proteins targeted to inclusion bodies.

    Science.gov (United States)

    Mačinković, Igor S; Abughren, Mohamed; Mrkic, Ivan; Grozdanović, Milica M; Prodanović, Radivoje; Gavrović-Jankulović, Marija

    2013-12-01

    High levels of recombinant protein expression can lead to the formation of insoluble inclusion bodies. These complex aggregates are commonly solubilized in strong denaturants, such as 6-8M urea, although, if possible, solubilization under milder conditions could facilitate subsequent refolding and purification of bioactive proteins. Commercially available GST-tag assays are designed for quantitative measurement of GST activity under native conditions. GST fusion proteins accumulated in inclusion bodies are considered to be undetectable by such assays. In this work, solubilization of recombinantly produced proteins was performed in 4M urea. The activity of rGST was assayed in 2M urea and it was shown that rGST preserves 85% of its activity under such denaturing conditions. A colorimetric GST activity assay with 1-chloro-2, 4-dinitrobenzene (CDNB) was examined for use in rapid detection of expression targeted to inclusion bodies and for the identification of inclusion body proteins which can be solubilized in low concentrations of chaotropic agents. Applicability of the assay was evaluated by tracking protein expression of two GST-fused allergens of biopharmaceutical value in E. coli, GST-Der p 2 and GST-Mus a 5, both targeted to inclusion bodies. Copyright © 2013 Elsevier B.V. All rights reserved.

  11. Influence of rhTPO/GM-CSF fusion protein on hemopoiesis in mice irradiated with 60Co γ-rays

    International Nuclear Information System (INIS)

    Cao Hua; Ge Zhongliang; Zhang Qunwei; Liu Xiuzhen

    1999-01-01

    Objective: To find a new biological therapy for secondary hematopoietic failure including anemia, infection and hemorrhage after administration of chemotherapeutic drugs etc. Methods: hGM-CSF gene was ligated with hTPO gene isolated from human fetal liver mRNA and a new fusion protein rh TPO/GM-CSF obtained. Results: The new fusion protein could promote recovery of peripheral WBC and PLT of 5.0 Gy irradiated mice. BFU-E, CFU-Meg and CFU-GM in bone marrow of mice after irradiation recovered significantly by treatment with rhTPO/GM-CSF fusion protein for 10 days. Conclusion: These results suggest that the new fusion protein has the biological activity of both hTPO and hGM-CSF simultaneously and can stimulate the proliferation of megakaryocytes and granulocyte progenitors

  12. Engineering design of the Nova Laser Facility for inertial-confinement fusion

    International Nuclear Information System (INIS)

    Simmons, W.W.; Godwin, R.O.; Hurley, C.A.

    1982-01-01

    The design of the Nova Laser Facility for inertial confinement fusion experiments at Lawrence Livermore National Laboratory is presented from an engineering perspective. Emphasis is placed upon design-to-performance requirements as they impact the various subsystems that comprise this complex experimental facility

  13. Design of intense neutron source for fusion material study and the role of universities

    International Nuclear Information System (INIS)

    Ishino, Shiori

    1993-01-01

    Need and requirement for the intense neutron source for fusion materials study have been discussed for many years. Recently, international climate has been becoming gradually maturing to consider this problem more seriously because of the recognition of crucial importance of solving materials problems for fusion energy development. The present symposium was designed to discuss the problems associated with the intense neutron source for material irradiation studies which will have a potential for the National Institute for Fusion Science to become one of the important future research areas. The symposium comprises five sessions; first, the role of materials research in fusion development strategies was discussed followed by a brief summary of current IFMIF (International Fusion Materials Irradiation Facility) activity. Despite the pressing need for intense fusion neutron source, currently available neutron sources are reactor or accelerator based sources of which FFTF and LASREF were discussed. Then, various concepts of intense neutron source candidates were presented including ESNIT, which are currently under design by JAERI. In the fourth session, discussions were made on the study of materials with the intense neutron source from the viewpoint of materials scientists and engineers as the user of the facility. This is followed by discussions on the role of universities from the two stand points, namely, fusion irradiation studies and fusion materials development. Finally summary discussions were made by the participants, indicating important role fundamental studies in universities for the full utilization of irradiation data and the need of pure 14 MeV neutron source for fundamental studies together with the intense surrogate neutron sources. (author)

  14. Probing plasma membrane microdomains in cowpea protoplasts using lipidated GFP-fusion proteins and multimode FRET microscopy

    NARCIS (Netherlands)

    Vermeer, J.E.M.; van Munster, E.B.; Vischer, N.O.; Gadella, T.

    2004-01-01

    Multimode fluorescence resonance energy transfer (FRET) microscopy was applied to study the plasma membrane organization using different lipidated green fluorescent protein (GFP)-fusion proteins co-expressed in cowpea protoplasts. Cyan fluorescent protein (CFP) was fused to the hyper variable region

  15. Functional role of the cytoplasmic tail domain of the major envelope fusion protein of group II baculoviruses

    NARCIS (Netherlands)

    Long, G.; Pan, M.; Westenberg, M.; Vlak, J.M.

    2006-01-01

    F proteins from baculovirus nucleopolyhedrovirus (NPV) group II members are the major budded virus (BV) viral envelope fusion proteins. They undergo furin-like proteolysis processing in order to be functional. F proteins from different baculovirus species have a long cytoplasmic tail domain (CTD),

  16. The destructive effect of botulinum neurotoxins on the SNARE protein: SNAP-25 and synaptic membrane fusion

    Directory of Open Access Journals (Sweden)

    Bin Lu

    2015-06-01

    Full Text Available Synaptic exocytosis requires the assembly of syntaxin 1A and SNAP-25 on the plasma membrane and synaptobrevin 2 (VAMP2 on the vesicular membrane to bridge the two opposite membranes. It is believed that the three SNARE proteins assemble in steps along the dynamic assembly pathway. The C-terminus of SNAP-25 is known to be the target of botulinum neurotoxins (BoNT/A and BoNT/E that block neurotransmitters release in vivo. In this study, we employed electron paramagnetic resonance (EPR spectroscopy to investigate the conformation of the SNAP-25 C-terminus in binary and ternary SNARE complexes. The fluorescence lipid mixing assay shows that the C-terminal of SNAP-25 is essential for membrane fusion, and that the truncated SNAP-25 mutants cleaved by BoNT/A and BoNT/E display different inhibition effects on membrane fusion: SNAP-25E (Δ26 abolishes the fusion activity of the SNARE complex, while SNAP-25A (Δ9 loses most of its function, although it can still form a SDS-resistant SNARE complex as the wild-type SNAP-25. CW-EPR spectra validate the unstable structures of the SNARE complex formed by SNAP-25 mutants. We propose that the truncated SNAP-25 mutants will disrupt the assembly of the SNARE core complex, and then inhibit the synaptic membrane fusion accordingly.

  17. Overview of the TITAN-II reversed-field pinch aqueous fusion power core design

    Energy Technology Data Exchange (ETDEWEB)

    Wong, C.P.C.; Creedon, R.L.; Grotz, S.; Cheng, E.T.; Sharafat, S.; Cooke, P.I.H.

    1988-03-01

    TITAN-II is a compact, high power density Reversed-Field Pinch fusion power reactor design based on the aqueous lithium solution fusion power core concept. The selected breeding and structural materials are LiNO/sub 3/ and 9-C low activation ferritic steel, respectively. TITAN-II is a viable alternative to the TITAN-I lithium self-cooled design for the Reversed-Field Pinch reactor to operate at a neutron wall loading of 18 MWm/sup 2/. Submerging the complete fusion power core and the primary loop in a large pool of cool water will minimize the probability of radioactivity release. Since the protection of the large pool integrity is the only requirement for the protection of the public, TITAN-II is a passive safety assurance design. 13 refs., 3 figs., 1 tab.

  18. Overview of the TITAN-II reversed-field pinch aqueous fusion power core design

    Energy Technology Data Exchange (ETDEWEB)

    Wong, C.P.C.; Creedon, R.L.; Cheng, E.T. (General Atomic Co., San Diego, CA (USA)); Grotz, S.P.; Sharafat, S.; Cooke, P.I.H. (California Univ., Los Angeles (USA). Dept. of Mechanical, Aerospace and Nuclear Engineering; California Univ., Los Angeles, CA (USA). Inst. for Plasma and Fusion Research); TITAN Research Group

    1989-04-01

    TITAN-II is a compact, high-power-density Reversed-Field Pinch fusion power reactor design based on the aqueous lithium solution fusion power core concept. The selected breeding and structural materials are LiNO/sub 3/ and 9-C low activation ferritic steel, respectively. TITAN-II is a viable alternative to the TITAN-I lithium self-cooled design for the Reversed-Field Pinch reactor to operate at a neutron wall loading of 18 MW/m/sup 2/. Submerging the complete fusion power core and the primary loop in a large pool of cool water will minimize the probability of radioactivity release. Since the protection of the large pool integrity is the only requirement for the protection of the public, TITAN-II is a level 2 of passive safety assurance design. (orig.).

  19. A study on the fusion reactor - A study on the design feature of fusion reactor divertor

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Kyung Jin [Chosun University, Kwangju (Korea, Republic of); Paek, Won Pil; Jang, Soon Hong [Korea Advanced Institute of Science and Technology, Taejon (Korea, Republic of); Sim, Young Jae [Kyungsang University, Jinju (Korea, Republic of)

    1996-09-01

    The contents and scope of the project can be summarized as, - study on the trend of divertor design - study on characteristics of coolant materials - study on characteristics of divertor materials - study on the thermal analysis method of divertor design. 36 refs., 12 tabs., 16 figs. (author)

  20. Present status of design, research and development of nuclear fusion reactors and problems

    International Nuclear Information System (INIS)

    1983-04-01

    Seven years have elapsed since the publication of ''Progress of nuclear fusion research and perspective toward the development of power reactors'' by the Atomic Energy Society of Japan in August, 1976. During this period, the research and development of nuclear fusion have changed from plasma physics to reactor technology, being conscious of the realization of fusion reactors. There are the R project in the Institute of Plasma Physics, Nagoya University, and the design and construction of JT-60 in Japan Atomic Energy Research Institute, to put it concretely. Now the research and development taking the economical efficiency into account are adopted. However, the type of fusion reactors is not reduced to tokamak type, accordingly the research and development to meet the diverse possibilities are forwarded. The progress of tokamak reactor research, core plasma design, nuclear design and shielding design, thermal structure design, the design of superconducting magnets, disassembling and repair, safety, economical efficiency, the conceptual design of other types than tokamak and others are reported. (Kako, I.)

  1. Recombinant protein blends: silk beyond natural design.

    Science.gov (United States)

    Dinjaski, Nina; Kaplan, David L

    2016-06-01

    Recombinant DNA technology and new material concepts are shaping future directions in biomaterial science for the design and production of the next-generation biomaterial platforms. Aside from conventionally used synthetic polymers, numerous natural biopolymers (e.g., silk, elastin, collagen, gelatin, alginate, cellulose, keratin, chitin, polyhydroxyalkanoates) have been investigated for properties and manipulation via bioengineering. Genetic engineering provides a path to increase structural and functional complexity of these biopolymers, and thereby expand the catalog of available biomaterials beyond that which exists in nature. In addition, the integration of experimental approaches with computational modeling to analyze sequence-structure-function relationships is starting to have an impact in the field by establishing predictive frameworks for determining material properties. Herein, we review advances in recombinant DNA-mediated protein production and functionalization approaches, with a focus on hybrids or combinations of proteins; recombinant protein blends or 'recombinamers'. We highlight the potential biomedical applications of fibrous protein recombinamers, such as Silk-Elastin Like Polypeptides (SELPs) and Silk-Bacterial Collagens (SBCs). We also discuss the possibility for the rationale design of fibrous proteins to build smart, stimuli-responsive biomaterials for diverse applications. We underline current limitations with production systems for these proteins and discuss the main trends in systems/synthetic biology that may improve recombinant fibrous protein design and production. Copyright © 2016. Published by Elsevier Ltd.

  2. Coating Nanoparticles with Plant-Produced Transferrin-Hydrophobin Fusion Protein Enhances Their Uptake in Cancer Cells

    DEFF Research Database (Denmark)

    Reuter, Lauri J.; Shahbazi, Mohammad-Ali; Makila, Ermei M.

    2017-01-01

    can be expressed in Nicotiana benthamiana plants as a fusion with Trichoderma reesei hydrophobins HFBI, HFBII, or HFBIV. Transferrin-HFBIV was further expressed in tobacco BY-2 suspension cells. Both partners of the fusion protein retained their functionality; the hydrophobin moiety enabled migration...... to a surfactant phase in an aqueous two-phase system, and the transferrin moiety was able to reversibly bind iron. Coating porous silicon nanoparticles with the fusion protein resulted in uptake of the nanoparticles in human cancer cells. This study provides a proof-of concept for the functionalization...

  3. Design study of electrical power supply system for tokamak fusion power reactor

    International Nuclear Information System (INIS)

    1977-01-01

    Design study of the electrical power supply system for a 2000MWt Tokamak-type fusion reactor has been carried out. The purposes are to reveal and study problems in the system, leading to a plan of the research and development. Performed were study of the electrical power supply system and design of superconducting inductive energy storages and power switches. In study of the system, specification and capability of various power supplies for the fusion power reactor and design of the total system with its components were investigated. For the superconducting inductive energy storages, material choice, design calculation, and structural design were conducted, giving the size, weight and performance. For thyristor switches, circuit design in the parallel / series connection of element valves and cooling design were studied, providing the size and weight. (auth.)

  4. Matrix protein 2 of influenza A virus blocks autophagosome fusion with lysosomes

    DEFF Research Database (Denmark)

    Gannagé, Monique; Dormann, Dorothee; Albrecht, Randy

    2009-01-01

    Influenza A virus is an important human pathogen causing significant morbidity and mortality every year and threatening the human population with epidemics and pandemics. Therefore, it is important to understand the biology of this virus to develop strategies to control its pathogenicity. Here, we...... demonstrate that influenza A virus inhibits macroautophagy, a cellular process known to be manipulated by diverse pathogens. Influenza A virus infection causes accumulation of autophagosomes by blocking their fusion with lysosomes, and one viral protein, matrix protein 2, is necessary and sufficient...... for this inhibition of autophagosome degradation. Macroautophagy inhibition by matrix protein 2 compromises survival of influenza virus-infected cells but does not influence viral replication. We propose that influenza A virus, which also encodes proapoptotic proteins, is able to determine the death of its host cell...

  5. Eradication of Human Hepatic and Pulmonary Melanoma Metastases in SCID Mice by Antibody--Interleukin 2 Fusion Proteins

    Science.gov (United States)

    Becker, Jurgen C.; Pancook, James D.; Gillies, Stephen D.; Mendelsohn, John; Reisfeld, Ralph A.

    1996-04-01

    Antibody--cytokine fusion proteins combine the unique targeting ability of antibodies with the multifunctional activity of cytokines. Here, we demonstrate the therapeutic efficacy of such constructs for the treatment of hepatic and pulmonary metastases of different melanoma cell lines. Two antibody--interleukin 2 (IL-2) fusion proteins, ch225-IL2 and ch14.18-IL2, constructed by fusion of a synthetic sequence coding for human IL-2 to the carboxyl end of the Cγ 1 gene of the corresponding antibodies, were tested for their therapeutic efficacy against xenografted human melanoma in vivo. Tumorspecific fusion proteins completely inhibited the growth of hepatic and pulmonary metastases in C.B-17 scid/scid mice previously reconstituted with human lymphokine-activated killer cells, whereas treatment with combinations of the corresponding antibodies plus recombinant IL-2 only reduced the tumor load. Even when treatment with fusion proteins was delayed up to 8 days after inoculation of tumor cells, it still resulted in complete eradication of micrometastases that were established at that time point. Selection of tumor cell lines expressing or lacking the targeted antigen of the administered fusion protein proved the specificity of the observed antitumor effect. Biodistribution analysis demonstrated that the tumorspecific fusion protein accumulated not only in subcutaneous tumors but also in lungs and livers affected with micrometastases. Survival times of animals treated with the fusion protein were more than doubled as compared to those treated with the combination of the corresponding antibody plus IL-2. Our data demonstrate that an immunotherapeutic approach using cytokines targeted by antibodies to tumor sites has potent effects against disseminated human melanoma.

  6. Osiris and SOMBRERO inertial fusion power plant designs - summary, conclusions, and recommendations

    International Nuclear Information System (INIS)

    Meier, Wayne R.

    1994-01-01

    An 18 month study to evaluate the potential of inertial fusion energy (IFE) for electric power production has been completed. The primary objective of the study was to provide the US Department of Energy with an evaluation of the potential of inertial fusion for electric power production. The study included the conceptual design of two inertial fusion power plants. Osiris uses an induction linac heavy ion beam driver, and SOMBRERO uses a krypton fluoride laser driver. Conceptual designs were completed for the reactors, power conversion and plant facilities, and drivers. Environmental and safety aspects, technical issues, technology development needs, and economics of the final point designs were assessed and compared. This paper summarizes the results and conclusions of the conceptual designs and results of the assessment studies. We conclude that IFE has the potential of producing technically credible designs with environmental, safety, and economics characteristics that are just as attractive as magnetic fusion. Realizing this potential will require additional research and development on target physics, chamber design, target production and injection systems, and drivers. ((orig.))

  7. A compact, multifunctional fusion module directs cholesterol-dependent homomultimerization and syncytiogenic efficiency of reovirus p10 FAST proteins.

    Directory of Open Access Journals (Sweden)

    Tim Key

    2014-03-01

    Full Text Available The homologous p10 fusion-associated small transmembrane (FAST proteins of the avian (ARV and Nelson Bay (NBV reoviruses are the smallest known viral membrane fusion proteins, and are virulence determinants of the fusogenic reoviruses. The small size of FAST proteins is incompatible with the paradigmatic membrane fusion pathway proposed for enveloped viral fusion proteins. Understanding how these diminutive viral fusogens mediate the complex process of membrane fusion is therefore of considerable interest, from both the pathogenesis and mechanism-of-action perspectives. Using chimeric ARV/NBV p10 constructs, the 36-40-residue ectodomain was identified as the major determinant of the differing fusion efficiencies of these homologous p10 proteins. Extensive mutagenic analysis determined the ectodomain comprises two distinct, essential functional motifs. Syncytiogenesis assays, thiol-specific surface biotinylation, and liposome lipid mixing assays identified an ∼25-residue, N-terminal motif that dictates formation of a cystine loop fusion peptide in both ARV and NBV p10. Surface immunofluorescence staining, FRET analysis and cholesterol depletion/repletion studies determined the cystine loop motif is connected through a two-residue linker to a 13-residue membrane-proximal ectodomain region (MPER. The MPER constitutes a second, independent motif governing reversible, cholesterol-dependent assembly of p10 multimers in the plasma membrane. Results further indicate that: (1 ARV and NBV homomultimers segregate to distinct, cholesterol-dependent microdomains in the plasma membrane; (2 p10 homomultimerization and cholesterol-dependent microdomain localization are co-dependent; and (3 the four juxtamembrane MPER residues present in the multimerization motif dictate species-specific microdomain association and homomultimerization. The p10 ectodomain therefore constitutes a remarkably compact, multifunctional fusion module that directs syncytiogenic

  8. Chemotropism and Cell Fusion in Neurospora crassa Relies on the Formation of Distinct Protein Complexes by HAM-5 and a Novel Protein HAM-14.

    Science.gov (United States)

    Jonkers, Wilfried; Fischer, Monika S; Do, Hung P; Starr, Trevor L; Glass, N Louise

    2016-05-01

    In filamentous fungi, communication is essential for the formation of an interconnected, multinucleate, syncytial network, which is constructed via hyphal fusion or fusion of germinated asexual spores (germlings). Anastomosis in filamentous fungi is comparable to other somatic cell fusion events resulting in syncytia, including myoblast fusion during muscle differentiation, macrophage fusion, and fusion of trophoblasts during placental development. In Neurospora crassa, fusion of genetically identical germlings is a highly dynamic and regulated process that requires components of a MAP kinase signal transduction pathway. The kinase pathway components (NRC-1, MEK-2 and MAK-2) and the scaffold protein HAM-5 are recruited to hyphae and germling tips undergoing chemotropic interactions. The MAK-2/HAM-5 protein complex shows dynamic oscillation to hyphae/germling tips during chemotropic interactions, and which is out-of-phase to the dynamic localization of SOFT, which is a scaffold protein for components of the cell wall integrity MAP kinase pathway. In this study, we functionally characterize HAM-5 by generating ham-5 truncation constructs and show that the N-terminal half of HAM-5 was essential for function. This region is required for MAK-2 and MEK-2 interaction and for correct cellular localization of HAM-5 to "fusion puncta." The localization of HAM-5 to puncta was not perturbed in 21 different fusion mutants, nor did these puncta colocalize with components of the secretory pathway. We also identified HAM-14 as a novel member of the HAM-5/MAK-2 pathway by mining MAK-2 phosphoproteomics data. HAM-14 was essential for germling fusion, but not for hyphal fusion. Colocalization and coimmunoprecipitation data indicate that HAM-14 interacts with MAK-2 and MEK-2 and may be involved in recruiting MAK-2 (and MEK-2) to complexes containing HAM-5. Copyright © 2016 by the Genetics Society of America.

  9. Interactions involved in pH protection of the alphavirus fusion protein

    Energy Technology Data Exchange (ETDEWEB)

    Fields, Whitney; Kielian, Margaret, E-mail: margaret.kielian@einstein.yu.edu

    2015-12-15

    The alphavirus membrane protein E1 mediates low pH-triggered fusion of the viral and endosome membranes during virus entry. During virus biogenesis E1 associates as a heterodimer with the transmembrane protein p62. Late in the secretory pathway, cellular furin cleaves p62 to the mature E2 protein and a peripheral protein E3. E3 remains bound to E2 at low pH, stabilizing the heterodimer and thus protecting E1 from the acidic pH of the secretory pathway. Release of E3 at neutral pH then primes the virus for fusion during entry. Here we used site-directed mutagenesis and revertant analysis to define residues important for the interactions at the E3–E2 interface. Our data identified a key residue, E2 W235, which was required for E1 pH protection and alphavirus production. Our data also suggest additional residues on E3 and E2 that affect their interacting surfaces and thus influence the pH protection of E1 during alphavirus exit.

  10. Protein Sub-Nuclear Localization Based on Effective Fusion Representations and Dimension Reduction Algorithm LDA

    Directory of Open Access Journals (Sweden)

    Shunfang Wang

    2015-12-01

    Full Text Available An effective representation of a protein sequence plays a crucial role in protein sub-nuclear localization. The existing representations, such as dipeptide composition (DipC, pseudo-amino acid composition (PseAAC and position specific scoring matrix (PSSM, are insufficient to represent protein sequence due to their single perspectives. Thus, this paper proposes two fusion feature representations of DipPSSM and PseAAPSSM to integrate PSSM with DipC and PseAAC, respectively. When constructing each fusion representation, we introduce the balance factors to value the importance of its components. The optimal values of the balance factors are sought by genetic algorithm. Due to the high dimensionality of the proposed representations, linear discriminant analysis (LDA is used to find its important low dimensional structure, which is essential for classification and location prediction. The numerical experiments on two public datasets with KNN classifier and cross-validation tests showed that in terms of the common indexes of sensitivity, specificity, accuracy and MCC, the proposed fusing representations outperform the traditional representations in protein sub-nuclear localization, and the representation treated by LDA outperforms the untreated one.

  11. Protein Sub-Nuclear Localization Based on Effective Fusion Representations and Dimension Reduction Algorithm LDA.

    Science.gov (United States)

    Wang, Shunfang; Liu, Shuhui

    2015-12-19

    An effective representation of a protein sequence plays a crucial role in protein sub-nuclear localization. The existing representations, such as dipeptide composition (DipC), pseudo-amino acid composition (PseAAC) and position specific scoring matrix (PSSM), are insufficient to represent protein sequence due to their single perspectives. Thus, this paper proposes two fusion feature representations of DipPSSM and PseAAPSSM to integrate PSSM with DipC and PseAAC, respectively. When constructing each fusion representation, we introduce the balance factors to value the importance of its components. The optimal values of the balance factors are sought by genetic algorithm. Due to the high dimensionality of the proposed representations, linear discriminant analysis (LDA) is used to find its important low dimensional structure, which is essential for classification and location prediction. The numerical experiments on two public datasets with KNN classifier and cross-validation tests showed that in terms of the common indexes of sensitivity, specificity, accuracy and MCC, the proposed fusing representations outperform the traditional representations in protein sub-nuclear localization, and the representation treated by LDA outperforms the untreated one.

  12. Botulinum neurotoxin: a marvel of protein design.

    Science.gov (United States)

    Montal, Mauricio

    2010-01-01

    Botulinum neurotoxin (BoNT), the causative agent of botulism, is acknowledged to be the most poisonous protein known. BoNT proteases disable synaptic vesicle exocytosis by cleaving their cytosolic SNARE (soluble NSF attachment protein receptor) substrates. BoNT is a modular nanomachine: an N-terminal Zn(2+)-metalloprotease, which cleaves the SNAREs; a central helical protein-conducting channel, which chaperones the protease across endosomes; and a C-terminal receptor-binding module, consisting of two subdomains that determine target specificity by binding to a ganglioside and a protein receptor on the cell surface and triggering endocytosis. For BoNT, functional complexity emerges from its modular design and the tight interplay between its component modules--a partnership with consequences that surpass the simple sum of the individual component's action. BoNTs exploit this design at each step of the intoxication process, thereby achieving an exquisite toxicity. This review summarizes current knowledge on the structure of individual modules and presents mechanistic insights into how this protein machine evolved to this level of sophistication. Understanding the design principles underpinning the function of such a dynamic modular protein remains a challenging task.

  13. Conceptual design study of fusion experimental reactor (FY86FER)

    International Nuclear Information System (INIS)

    Nakashima, Kunihiko; Ishigaki, Yukio; Ozaki, Akira; Yamane, Minoru.

    1987-09-01

    This report describes the results of the capacity estimation for the electrical power system on the typical two candidates for the FER (Fusion Experimental Reactor) which were picked out through the process of '86 FER scoping studies. Main concern in the electrical systems is coil power supplies which have a capacity of about 1 GW, and this is dominated by poloidal coil power supplies. Then, studies to reduce the converter capacity are concentrated on the poloidal coil power system in relation to the sypplying poloidal flux at the initial phase of plasma ramp-up. A quench protection circuit was proposed on the toroidal coil power supply. On the position control power supply, a circuit with reasonable functions was proposed. Under these system studies, general specifications were determined and the capacity of each power supply unit was estimated. On the poloidal coil power supply system, the accumulated capacity of converters amounted to 885 MW for the one candidate and 782 MW for another. (author)

  14. Conceptual design study of fusion experimental reactor (FY86 FER)

    International Nuclear Information System (INIS)

    Miki, Nobuharu; Iida, Fumio; Suzuki, Shohei; Wachi, Yoshihiro; Toyoda, Katsuyoshi; Hashizume, Takashi; Konno, Masayuki.

    1987-09-01

    This report summarizes the FER magnet design which was conducted last year (1986). Main objective of the new FER design is to have better cost performance of the machine. The physics assumptions are reviewed to reduce risks. Optimization of the physics design and improvements of the engineering design have been done without changing missions of the device. After a preliminary investigation for the optimization and improvements, six FER concepts have been developed to establish the improved design point, and have been studied in more detail. In the magnet design, the improvements of superconducting magnet design were mainly investigated to reduce the reactor size. A normal conductor was studied as an alternative option for appling to the special poloidal field coils that were located on the interior to the toroidal field coils. Some improvements were made on the superconducting magnet design. Based on the preliminary investigation, the magnet design specifications have been modified somewhat. The conceptual design of the magnet system components have been done for the candidate FER concepts. (author)

  15. Design assumptions and bases for small D-T-fueled Sperical Tokamak (ST) fusion core

    International Nuclear Information System (INIS)

    Peng, Y.K.M.; Galambos, J.D.; Fogarty, P.J.

    1996-01-01

    Recent progress in defining the assumptions and clarifying the bases for a small D-T-fueled ST fusion core are presented. The paper covers several issues in the physics of ST plasmas, the technology of neutral beam injection, the engineering design configuration, and the center leg material under intense neutron irradiation. This progress was driven by the exciting data from pioneering ST experiments, a heightened interest in proof-of-principle experiments at the MA level in plasma current, and the initiation of the first conceptual design study of the small ST fusion core. The needs recently identified for a restructured fusion energy sciences program have provided a timely impetus for examining the subject of this paper. Our results, though preliminary in nature, strengthen the case for the potential realism and attractiveness of the ST approach

  16. Enhanced neutralization potency of botulinum neurotoxin antibodies using a red blood cell-targeting fusion protein.

    Directory of Open Access Journals (Sweden)

    Sharad P Adekar

    2011-03-01

    Full Text Available Botulinum neurotoxin (BoNT potently inhibits cholinergic signaling at the neuromuscular junction. The ideal countermeasures for BoNT exposure are monoclonal antibodies or BoNT antisera, which form BoNT-containing immune complexes that are rapidly cleared from the general circulation. Clearance of opsonized toxins may involve complement receptor-mediated immunoadherence to red blood cells (RBC in primates or to platelets in rodents. Methods of enhancing immunoadherence of BoNT-specific antibodies may increase their potency in vivo. We designed a novel fusion protein (FP to link biotinylated molecules to glycophorin A (GPA on the RBC surface. The FP consists of an scFv specific for murine GPA fused to streptavidin. FP:mAb:BoNT complexes bound specifically to the RBC surface in vitro. In a mouse model of BoNT neutralization, the FP increased the potency of single and double antibody combinations in BoNT neutralization. A combination of two antibodies with the FP gave complete neutralization of 5,000 LD50 BoNT in mice. Neutralization in vivo was dependent on biotinylation of both antibodies and correlated with a reduction of plasma BoNT levels. In a post-exposure model of intoxication, FP:mAb complexes gave complete protection from a lethal BoNT/A1 dose when administered within 2 hours of toxin exposure. In a pre-exposure prophylaxis model, mice were fully protected for 72 hours following administration of the FP:mAb complex. These results demonstrate that RBC-targeted immunoadherence through the FP is a potent enhancer of BoNT neutralization by antibodies in vivo.

  17. Intracellular delivery of cell-penetrating peptide-transcriptional factor fusion protein and its role in selective osteogenesis

    Science.gov (United States)

    Suh, Jin Sook; Lee, Jue Yeon; Choi, Yoon Jung; You, Hyung Keun; Hong, Seong-Doo; Chung, Chong Pyoung; Park, Yoon Jeong

    2014-01-01

    Protein-transduction technology has been attempted to deliver macromolecular materials, including protein, nucleic acids, and polymeric drugs, for either diagnosis or therapeutic purposes. Herein, fusion protein composed of an arginine-rich cell-penetrating peptide, termed low-molecular-weight protamine (LMWP), and a transcriptional coactivator with a PDZ-binding motif (TAZ) protein was prepared and applied in combination with biomaterials to increase bone-forming capacity. TAZ has been recently identified as a specific osteogenic stimulating transcriptional coactivator in human mesenchymal stem cell (hMSC) differentiation, while simultaneously blocking adipogenic differentiation. However, TAZ by itself cannot penetrate the cells, and thus needs a transfection tool for translocalization. The LMWP-TAZ fusion proteins were efficiently translocalized into the cytosol of hMSCs. The hMSCs treated with cell-penetrating LMWP-TAZ exhibited increased expression of osteoblastic genes and protein, producing significantly higher quantities of mineralized matrix compared to free TAZ. In contrast, adipogenic differentiation of the hMSCs was blocked by treatment of LMWP-TAZ fusion protein, as reflected by reduced marker-protein expression, adipocyte fatty acid-binding protein 2, and peroxisome proliferator-activated receptor-γ messenger ribonucleic acid levels. LMWP-TAZ was applied in alginate gel for the purpose of localization and controlled release. The LMWP-TAZ fusion protein-loaded alginate gel matrix significantly increased bone formation in rabbit calvarial defects compared with alginate gel matrix mixed with free TAZ protein. The protein transduction of TAZ fused with cell-penetrating LMWP peptide was able selectively to stimulate osteogenesis in vitro and in vivo. Taken together, this fusion protein-transduction technology for osteogenic protein can thus be applied in combination with biomaterials for tissue regeneration and controlled release for tissue

  18. Intrafocal heterogeneity of ERG protein expression and gene fusion pattern in prostate cancer.

    Science.gov (United States)

    Suh, Ja Hee; Park, Jeong Hwan; Lee, Cheol; Moon, Kyung Chul

    2017-10-01

    Prostate cancer is considered to be highly heterogeneous, with various morphologic features and biologic behaviors. The TMPRSS2-ERG gene fusion is the most frequently observed genetic aberration in prostate cancer. The aim of this study was to elucidate the intrafocal heterogeneity of ERG gene fusion status. ERG immunohistochemistry (IHC) was performed in samples from 168 prostate cancer patients who had undergone radical prostatectomy, and 40 cases showing ERG-positive IHC staining were selected for tissue microarray (TMA) construction. Two to six representative cores were selected from each tumor focus. In the cases with heterogeneous ERG IHC staining intensity, the areas showing different intensities were separately selected. Using the TMA blocks, IHC and fluorescence in situ hybridization (FISH) were conducted to evaluate the heterogeneity of ERG protein expression and ERG fusion gene patterns, respectively, in a single tumor focus. Heterogeneity of ERG IHC staining was defined as the simultaneous presence of negative and positive cores in the same tumor focus. Heterogeneity of ERG FISH was defined by the presence of cores with positive and negative FISH signals or cores with break-apart and interstitial deletion FISH signals in the same tumor focus. A total of 202 TMA cores were isolated from 40 ERG-positive cases. Of the 202 total cores, 19 were negative for ERG IHC staining, and 46 showed 1+, 52 showed 2+, and 85 showed 3+ ERG staining intensity. Eleven cores were negative for ERG FISH signal, 119 cores showed ERG break-apart FISH signals, and the remaining 72 cores revealed interstitial deletion. Intrafocal heterogeneity of ERG IHC staining was found in 20% (8/40) of cases, and intrafocal heterogeneity of ERG gene fusion pattern was found in 32.5% (13/40) of cases. In summary, this study showed significantly frequent intrafocal heterogeneity of ERG protein expression, gene fusion status and fusion pattern. This heterogeneity can be caused by the development

  19. Once for All: A Novel Robust System for Co-expression of Multiple Chimeric Fluorescent Fusion Proteins in Plants

    Directory of Open Access Journals (Sweden)

    Guitao Zhong

    2017-06-01

    Full Text Available Chimeric fluorescent fusion proteins have been employed as a powerful tool to reveal the subcellular localizations and dynamics of proteins in living cells. Co-expression of a fluorescent fusion protein with well-known organelle markers in the same cell is especially useful in revealing its spatial and temporal functions of the protein in question. However, the conventional methods for co-expressing multiple fluorescent tagged proteins in plants have the drawbacks of low expression efficiency, variations in the expression level and time-consuming genetic crossing. Here, we have developed a novel robust system that allows for high-efficient co-expression of multiple chimeric fluorescent fusion proteins in plants in a time-saving fashion. This system takes advantage of employing a single expression vector which consists of multiple semi-independent expressing cassettes for the protein co-expression thereby overcoming the limitations of using multiple independent expressing plasmids. In addition, it is a highly manipulable DNA assembly system, in which modification and recombination of DNA molecules are easily achieved through an optimized one-step assembly reaction. By employing this effective system, we demonstrated that co-expression of two chimeric fluorescent fusion reporter proteins of vacuolar sorting receptor and secretory carrier membrane protein gave rise to their perspective subcellular localizations in plants via both transient expression and stable transformation. Thus, we believed that this technical advance represents a promising approach for multi-color-protein co-expression in plant cells.

  20. Expression, purification, crystallization and preliminary X-ray crystallographic studies of hepatitis B virus core fusion protein corresponding to octahedral particles

    International Nuclear Information System (INIS)

    Kikuchi, Masaki; Iwabuchi, Shinichiro; Kikkou, Tatsuhiko; Noguchi, Keiichi; Odaka, Masafumi; Yohda, Masafumi; Kawata, Masaaki; Sato, Chikara; Matsumoto, Osamu

    2013-01-01

    Novel hepatitis B virus-like particles of recombinant dimeric core–GFP fusion protein were expressed, purified and crystallized. The crystals diffracted to 2.15 Å resolution and belonged to space group F432, with unit-cell parameters a = b = c = 219.7 Å. Recombinant hepatitis B virus core proteins dimerize to form building blocks that are capable of self-assembly into a capsid. A core capsid protein dimer (CPD) linked to a green fluorescent protein variant, EGFP, at the C-terminus has been designed. The recombinant fusion CPD was expressed in Escherichia coli, assembled into virus-like particles (VLPs), purified and crystallized. The single crystal diffracted to 2.15 Å resolution and belonged to the cubic space group F432, with unit-cell parameters a = b = c = 219.7 Å. The fusion proteins assembled into icosahedral VLPs in aqueous solution, but were rearranged into octahedral symmetry through the crystal-packing process under the crystallization conditions

  1. Conceptual design considerations and neutronics of lithium fall laser fusion target chambers

    International Nuclear Information System (INIS)

    Meier, W.R.; Thomson, W.B.

    1978-01-01

    Atomics International and Lawrence Livermore Laboratory are involved in the conceptual design of a laser fusion power plant incorporating the lithium fall target chamber. In this paper we discuss some of the more important design considerations for the target chamber and evaluate its nuclear performance. Sizing and configuration of the fall, hydraulic effects, and mechanical design considerations are addressed. The nuclear aspects examined include tritium breeding, energy deposition, and radiation damage

  2. Prm3p is a pheromone-induced peripheral nuclear envelope protein required for yeast nuclear fusion.

    Science.gov (United States)

    Shen, Shu; Tobery, Cynthia E; Rose, Mark D

    2009-05-01

    Nuclear membrane fusion is the last step in the mating pathway of the yeast Saccharomyces cerevisiae. We adapted a bioinformatics approach to identify putative pheromone-induced membrane proteins potentially required for nuclear membrane fusion. One protein, Prm3p, was found to be required for nuclear membrane fusion; disruption of PRM3 caused a strong bilateral defect, in which nuclear congression was completed but fusion did not occur. Prm3p was localized to the nuclear envelope in pheromone-responding cells, with significant colocalization with the spindle pole body in zygotes. A previous report, using a truncated protein, claimed that Prm3p is localized to the inner nuclear envelope. Based on biochemistry, immunoelectron microscopy and live cell microscopy, we find that functional Prm3p is a peripheral membrane protein exposed on the cytoplasmic face of the outer nuclear envelope. In support of this, mutations in a putative nuclear localization sequence had no effect on full-length protein function or localization. In contrast, point mutations and deletions in the highly conserved hydrophobic carboxy-terminal domain disrupted both protein function and localization. Genetic analysis, colocalization, and biochemical experiments indicate that Prm3p interacts directly with Kar5p, suggesting that nuclear membrane fusion is mediated by a protein complex.

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

    Science.gov (United States)

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

    2017-01-01

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

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

    Directory of Open Access Journals (Sweden)

    Fatemeh Shafiee

    2017-01-01

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

  5. Conceptual design study of Fusion Experimental Reactor (FY87FER)

    International Nuclear Information System (INIS)

    Miki, Nobuharu; Iida, Fumio; Wachi, Yoshihiro; Toyoda, Katsuyoshi; Hashizume, Takashi; Konno, Masayuki.

    1988-06-01

    This report describes the FER magnet design which was conducted last year (1987). Based on a large uncertainty of the physics assumption, two sets of FER concepts have been developed. One is based on the best existing physics data bases and another is based on rather conservative physics bases. In the magnet design, the improvements of superconducting magnet design were investigated to reduce the reactor size and to realize higher reactor-core performance. In addition, we studied several critical technical issues that affect the magnet design specification. (author)

  6. Parametric design study of tandem mirror fusion reactors

    International Nuclear Information System (INIS)

    Carlson, G.A.

    1977-01-01

    The parametric design study of the tandem mirror reactor (TMR) is described. The results of this study illustrate the variation of reactor characteristics with changes in the independent design parameters, reveal the set of design parameters which minimizes the cost of the reactor, and show the sensitivity of the optimized design to physics and technological uncertainties. The total direct capital cost of an optimized 1000 MWe TMR is estimated to be $1300/kWe. The direct capital cost of a 2000 MWe plant is less than $1000/kWe

  7. Synergies in the design and development of fusion and generation IV fission reactors

    International Nuclear Information System (INIS)

    Bogusch, E.; Carre, F.; Knebel, J.; Aoto, K.

    2007-01-01

    Future fusion reactors or systems and Generation IV fission reactors are designed and developed in worldwide programmes mostly involving the same partners to investigate and assess their potential for realisation and contribution to meet the future energy needs beyond 2030. Huge scientific and financial effort is necessary to meet these objectives. First programmes have been launched in Generation IV International Forum (GIF) for fission and in the Broader Approach for fusion reactor system development. Except the physics basis for the energy source, future fusion and fission reactors, in particular those with fast neutron core face similar design issues and development needs. Therefore the call for the identification of synergies became evident. Beyond ITER cooled by water, future fusion reactors or systems will be designed for helium and liquid metal cooling and higher temperatures similar to those proposed for some of the six fission reactor concepts in GIF with their diverse coolants. Beside materials developments which are not discussed in this paper, design and performance of components and systems related to the diverse coolants including lifetime and maintenance aspects might offer significant potentials for synergies. Furthermore, the use of process heat for applications in addition to electricity production as well as their safety approaches might create synergistic design and development programmes. Therefore an early identification of possible synergies in the relevant programmes should be endorsed to minimise the effort for future power plants in terms of investments and resources. In addition to a general overview of a possible synergistic work programme which promotes the interaction between fusion and fission programmes towards an integrated organisation of their design and R and D programmes, some specific remarks will be given for joint design tools, numerical code systems and joint experiments in support of common technologies. (orig.)

  8. Synergies in the design and development of fusion and generation IV fission reactors

    International Nuclear Information System (INIS)

    Bogusch, E.; Carre, F.; Knebel, J.U.; Aoto, K.

    2008-01-01

    Future fusion reactor and Generation IV fission reactor systems are designed and developed in worldwide programmes to investigate and assess their potential for realisation and contribution to the future energy needs beyond 2030 mostly involving the same partners. Huge scientific and financial effort is necessary to meet these objectives. First programmes have been launched in Generation IV International Forum (GIF) for fission and in the Broader Approach for fusion reactor system development. Except for the physics basis for the energy source, future fusion and fission reactors, in particular those with fast neutron core, face similar design issues and development needs. Therefore, the call for the identification of synergies became evident. Beyond ITER cooled by water, future fusion reactor systems will be designed for high-temperature helium and liquid metal cooling but also water including supercritical water and molten salt similar to those proposed for some of the six fission reactor concepts in GIF with their diverse coolants. Beside materials developments which are not discussed in this paper, design and performance of components and systems related to the diverse coolants including lifetime and maintenance aspects might offer significant potentials for synergies. Furthermore, the use of process heat for applications in addition to electricity production as well as their safety approaches can create synergistic design and development programmes. Therefore, an early identification of possible synergies in the relevant programmes should be endorsed to minimise the effort for future power plants in terms of investments and resources. In addition to a general overview of a possible synergistic work programme which promotes the interaction between fusion and fission programmes towards an integrated organisation of their design and R and D programmes, some specific remarks will be given for joint design tools, numerical code systems and joint experiments in

  9. Preconceptual design of hyfire. A fusion driven high temperature electrolysis plant

    International Nuclear Information System (INIS)

    Varljen, T.C.; Chi, J.W.H.; Karbowski, J.S.

    1983-01-01

    Brookhaven National Laboratory has been engaged in a scoping study to investigate the potential merits of coupling a fusion reactor with a high temperature blanket to a high temperature electrolysis (HTE) process to produce hydrogen and oxygen. Westinghouse is assisting this study in the areas of systems design integration, plasma engineering, balance of plant design and electrolyzer technology. The aim of the work done in the past year has been to focus on a reference design point for the plant, which has been designated HYFIRE. In prior work, the STARFIRE commercial tokamak fusion reactor was directly used as the fusion driver. This report describes a new design obtained by scaling the basic STARFIRE design to permit the achievement of a blanket power of 6000 MWt. The high temperature blanket design employs a thermally insulated refractory oxide region which provides high temperature (>1000 deg. C) steam at moderate pressures to high temperature electrolysis units. The electrolysis process selected is based on the high temperature, solid electrolyte fuel cell technology developed by Westinghouse. An initial process design and plant layout has been completed; component cost and plant economics studies are now underway to develop estimates of hydrogen production costs and to determine the sensitivity of this cost to changes in major design parameters. (author)

  10. Public acceptance of fusion energy and scientific feasibility of a fusion reactor. Design of inductively driven long pulse tokamak reactors: IDLT

    International Nuclear Information System (INIS)

    Ogawa, Yuichi

    1998-01-01

    Based on scientific data based adopted for designing ITER plasmas and on the advancement of fusion nuclear technology from the recent R and D program, the scientific feasibility of inductively-driven tokamak fusion reactors is studied. A low wall-loading DEMO fusion reactor is designed, which utilizes an austenitic stainless steel in conjunction with significant data bases and operating experiences, since we have given high priority to the early and reliable realization of a tokamak fusion plasma over the cost performance. Since the DEMO reactor with the relatively large volume (i.e., major radius of 10 m) is employed, plasma ignition is achievable with a low fusion power of 0.8 GW, and an operation period of 4 - 5 hours is available only with inductive current drive. Disadvantages of pulsed operation in commercial fusion reactors include fatigue in structural materials and the necessity of an energy storage system to compensate the electric power during the dwell time. To overcome these disadvantages, a pulse length is prolonged up to about 10 hours, resulting in the remarkable reduction of the total cycle number to 10 4 during the life of the fusion plant. (author)

  11. Safety design of the international fusion materials irradiation facility (IFMIF)

    International Nuclear Information System (INIS)

    Konishi, Satoshi; Yamaki, Daiju; Katsuta, Hiroji; Moeslang, Anton; Jameson, R.A.; Martone, Marcello; Shannon, T.E.

    1997-11-01

    In the Conceptual Design Activity of the IFMIF, major subsystems, as well as the entire facility is carefully designed to satisfy the safety requirements for any possible construction sites. Each subsystem is qualitatively analyzed to identify possible hazards to the workers, public and environments using Failure Mode and Effect Analysis (FMEA). The results are reflected in the design and operation procedure. Shielding of radiation, particularly neutron around the test cell is one of the most important issue in normal operation. Radiation due to beam halo and activation is a hazard for operation personnel in the accelerator system. For the maintenance, remote handling technology is designed to be applied in various facilities of the IFMIF. Lithium loop and target system hold the majority of the radioactive material in the facility. Tritium and beryllium-7 are generated by the nuclear reaction during operation and thus needed to be removed continuously. They are also the potential hazards of airborne source in off-normal events. Minimization of inventory, separation and immobilization, and multiple confinement are considered in the design. Generation of radioactive waste is anticipated to be minor, but waste treatment systems for gas, liquid and solid wastes are designed to minimize the environmental impact. Lithium leak followed by a fire is a major concern, and extensive prevention plan is made in the target design. One of the design option considered is composed of; primary enclosure of the lithium loop, secondary containment filled with positive pressure argon, and an air tight lithium cell made of concrete with a steel lining. This study will report some technical issues considered in the design of IFMIF. It was concluded that the IFMIF can be designed and constructed to meet or exceed current safely standards for workers, public and the environment with existing technology and reasonable construction cost. (J.P.N.)

  12. Fcγ1 fragment of IgG1 as a powerful affinity tag in recombinant Fc-fusion proteins: immunological, biochemical and therapeutic properties.

    Science.gov (United States)

    Soleimanpour, Saman; Hassannia, Tahereh; Motiee, Mahdieh; Amini, Abbas Ali; Rezaee, S A R

    2017-05-01

    Affinity tags are vital tools for the production of high-throughput recombinant proteins. Several affinity tags, such as the hexahistidine tag, maltose-binding protein, streptavidin-binding peptide tag, calmodulin-binding peptide, c-Myc tag, glutathione S-transferase and FLAG tag, have been introduced for recombinant protein production. The fragment crystallizable (Fc) domain of the IgG1 antibody is one of the useful affinity tags that can facilitate detection, purification and localization of proteins and can improve the immunogenicity, modulatory effects, physicochemical and pharmaceutical properties of proteins. Fcγ recombinant forms a group of recombinant proteins called Fc-fusion proteins (FFPs). FFPs are widely used in drug discovery, drug delivery, vaccine design and experimental research on receptor-ligand interactions. These fusion proteins have become successful alternatives to monoclonal antibodies for drug developments. In this review, the physicochemical, biochemical, immunological, pharmaceutical and therapeutic properties of recombinant FFPs were discussed as a new generation of bioengineering strategies.

  13. Conceptual design of a laser fusion power plant. Part I. An integrated facility

    International Nuclear Information System (INIS)

    1981-07-01

    This study is a new preliminary conceptual design and economic analysis of an inertial confinement fusion (ICF) power plant performed by Bechtel under the direction of Lawrence Livermore National Laboratory (LLNL). The purpose of a new conceptual design is to examine alternatives to the LLNL HYLIFE power plant and to incorporate information from the recent liquid metal cooled power plant conceptual design study (CDS) into the reactor system and balance of plant design. A key issue in the design of a laser fusion power plant is the degree of symmetry in the illumination of the target that will be required for a proper burn. Because this matter is expected to remain unresolved for some time, another purpose of this study is to determine the effect of symmetry requirements on the total plant size, layout, and cost

  14. Conceptual design of blanket structures for fusion experimental reactor (FER)

    International Nuclear Information System (INIS)

    1984-03-01

    Conceptual design study for in-vessel components including tritium breeding blanket of FER has been carried out. The objective of this study is to obtain the engineering and technological data for selecting the reactor concept and for its construction by investigating fully and broadly. The design work covers in-vessel components (such as tritium breeding blanket, first wall, shield, divertor and blanket test module), remote handling system and tritium system. The designs of those components and systems are accomplished in consideration of their accomodation to whole reactor system and problems for furthur study are clarified. (author)

  15. Conceptual design of a fast-ignition laser fusion reactor FALCON-D

    International Nuclear Information System (INIS)

    Goto, T.; Ogawa, Y.; Okano, K.; Hiwatari, R.; Asaoka, Y.; Someya, Y.; Sunahara, A.; Johzaki, T.

    2008-10-01

    A new conceptual design of the laser fusion power plant FALCON-D (Fast ignition Advanced Laser fusion reactor CONcept with a Dry wall chamber) has been proposed. The fast ignition method can achieve the sufficient fusion gain for a commercial operation (∼100) with about 10 times smaller fusion yield than the conventional central ignition method. FALCON-D makes full use of this property and aims at designing with a compact dry wall chamber (5 - 6 m radius). 1-D/2-D hydrodynamic simulations showed the possibility of the sufficient gain achievement with a 40 MJ target yield. The design feasibility of the compact dry wall chamber and solid breeder blanket system was shown through the thermomechanical analysis of the dry wall and neutronics analysis of the blanket system. A moderate electric output (∼400 MWe) can be achieved with a high repetition (30 Hz) laser. This dry wall concept not only reduces some difficulties accompanied with a liquid wall but also enables a simple cask maintenance method for the replacement of the blanket system, which can shorten the maintenance time. The basic idea of the maintenance method for the final optics system has also been proposed. Some critical R and D issues required for this design are also discussed. (author)

  16. Space-time energy concentration and the design of DT fusion micro-explosions

    International Nuclear Information System (INIS)

    Sahlin, H.L.; Brandenburg, J.E.

    1980-03-01

    As part of the effort to employ a plasma focus as a driver for fusion micro-explosions, many target concepts were explored and extensive imposion calculations have been carried out. Some of the basic principles of micro-explosion design are presented

  17. Human cytomegaloviruses expressing yellow fluorescent fusion proteins--characterization and use in antiviral screening.

    Directory of Open Access Journals (Sweden)

    Sarah Straschewski

    Full Text Available Recombinant viruses labelled with fluorescent proteins are useful tools in molecular virology with multiple applications (e.g., studies on intracellular trafficking, protein localization, or gene activity. We generated by homologous recombination three recombinant cytomegaloviruses carrying the enhanced yellow fluorescent protein (EYFP fused with the viral proteins IE-2, ppUL32 (pp150, and ppUL83 (pp65. In growth kinetics, the three viruses behaved all like wild type, even at low multiplicity of infection (MOI. The expression of all three fusion proteins was detected, and their respective localizations were the same as for the unmodified proteins in wild-type virus-infected cells. We established the in vivo measurement of fluorescence intensity and used the recombinant viruses to measure inhibition of viral replication by neutralizing antibodies or antiviral substances. The use of these viruses in a pilot screen based on fluorescence intensity and high-content analysis identified cellular kinase inhibitors that block viral replication. In summary, these viruses with individually EYFP-tagged proteins will be useful to study antiviral substances and the dynamics of viral infection in cell culture.

  18. Fusion protein-based biofilm fabrication composed of recombinant azurin–myoglobin for dual-level biomemory application

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Taek [Research Institute for Basic Science, Sogang University, Heukseok-dong, Dongjak-gu, Seoul 156-756 (Korea, Republic of); Chung, Yong-Ho; Yoon, Jinho [Department of Chemical and Biomolecular Engineering, Sogang University, Heukseok-dong, Dongjak-gu, 35 Baekbeom-ro (Sinsu-dong), Mapo-gu, Seoul 121-742 (Korea, Republic of); Min, Junhong [School of Integrative Engineering, Chung-Ang University, Heukseok-dong, Dongjak-gu, Seoul 156-756 (Korea, Republic of); Choi, Jeong-Woo, E-mail: jwchoi@sogang.ac.kr [Department of Chemical and Biomolecular Engineering, Sogang University, Heukseok-dong, Dongjak-gu, 35 Baekbeom-ro (Sinsu-dong), Mapo-gu, Seoul 121-742 (Korea, Republic of)

    2014-11-30

    Graphical abstract: - Highlights: • We developed the fusion protein-based biofilm on the inorganic surface. • For making the fusion protein, the recombinant azurin and the myoglobin was conjugated by the native chemical ligation method. • The developed fusion protein shows unique electrochemical property. • The proposed fusion protein biofilm appears to be a good method for dual-level biomemory device. - Abstract: In the present study, a fusion protein-based biofilm composed of a recombinant azurin–myoglobin (Azu-Myo) has been developed and confirmed its original electrochemical property for dual-level biomemory device application. For this purpose, the azurin was modified with cysteine residues for direct immobilization and conjugation. Then, the recombinant azurin was conjugated with the myoglobin via a sulfo-SMCC bifunctional linker using the chemical ligation method (CLM). The SDS-PAGE and UV–vis spectroscopy were performed to examine the fusion protein conjugates. The prepared Azu-Myo fusion protein was self-assembled onto Au substrate for the biofilm fabrication. Then, the atomic force microscopy (AFM) was used to confirm the immobilization and the surface-enhanced Raman spectroscopy (SERS) was carried out to the surface analysis. Also, the cyclic voltammetry (CV) was carried out to observe an electrochemical property of fabricated biofilm. As a result, the two pair of redox potential values was obtained for dual-level biomemory device application. Then, the dual-level biomemory function was verified by the multi-potential chronoamperometry (MPCA). The results indicate a new fabrication method and material combination for advances in bioelectronic device development.

  19. Fusion protein-based biofilm fabrication composed of recombinant azurin–myoglobin for dual-level biomemory application

    International Nuclear Information System (INIS)

    Lee, Taek; Chung, Yong-Ho; Yoon, Jinho; Min, Junhong; Choi, Jeong-Woo

    2014-01-01

    Graphical abstract: - Highlights: • We developed the fusion protein-based biofilm on the inorganic surface. • For making the fusion protein, the recombinant azurin and the myoglobin was conjugated by the native chemical ligation method. • The developed fusion protein shows unique electrochemical property. • The proposed fusion protein biofilm appears to be a good method for dual-level biomemory device. - Abstract: In the present study, a fusion protein-based biofilm composed of a recombinant azurin–myoglobin (Azu-Myo) has been developed and confirmed its original electrochemical property for dual-level biomemory device application. For this purpose, the azurin was modified with cysteine residues for direct immobilization and conjugation. Then, the recombinant azurin was conjugated with the myoglobin via a sulfo-SMCC bifunctional linker using the chemical ligation method (CLM). The SDS-PAGE and UV–vis spectroscopy were performed to examine the fusion protein conjugates. The prepared Azu-Myo fusion protein was self-assembled onto Au substrate for the biofilm fabrication. Then, the atomic force microscopy (AFM) was used to confirm the immobilization and the surface-enhanced Raman spectroscopy (SERS) was carried out to the surface analysis. Also, the cyclic voltammetry (CV) was carried out to observe an electrochemical property of fabricated biofilm. As a result, the two pair of redox potential values was obtained for dual-level biomemory device application. Then, the dual-level biomemory function was verified by the multi-potential chronoamperometry (MPCA). The results indicate a new fabrication method and material combination for advances in bioelectronic device development

  20. Design issues and implications for the structural integrity and lifetime of fusion power plant components

    International Nuclear Information System (INIS)

    Karditas, P.J.

    1996-05-01

    This review discusses, with example calculations, the criteria, and imposed constraints and limitations, for the design of fusion components and assesses the implications for successful design and power plant operation. The various loading conditions encountered during the operation of a tokamak lead to structural damage and possible failure by such mechanisms as yielding, thermal creep rupture and fatigue due to thermal cycling, plastic strain cycling (ratcheting), crack growth-propagation and radiation induced swelling and creep. Of all the possible damage mechanisms, fatigue, creep and their combination are the most important in the structural design and lifetime of fusion power plant components operating under steady or load varying conditions. Also, the effect of neutron damage inflicted onto the structural materials and the degradation of key properties is of major concern in the design and lifetime prediction of components. Structures are classified by, and will be restricted by existing or future design codes relevant to medium and high temperature power plant environments. The ways in which existing design codes might be used in present and near future design activities, and the implications, are discussed; the desirability of an early start towards the development of fusion-specific design codes is emphasised. (UK)

  1. Nuclear-thermal-coupled optimization code for the fusion breeding blanket conceptual design

    International Nuclear Information System (INIS)

    Li, Jia; Jiang, Kecheng; Zhang, Xiaokang; Nie, Xingchen; Zhu, Qinjun; Liu, Songlin

    2016-01-01

    Highlights: • A nuclear-thermal-coupled predesign code has been developed for optimizing the radial build arrangement of fusion breeding blanket. • Coupling module aims at speeding up the efficiency of design progress by coupling the neutronics calculation code with the thermal-hydraulic analysis code. • Radial build optimization algorithm aims at optimal arrangement of breeding blanket considering one or multiple specified objectives subject to the design criteria such as material temperature limit and available TBR. - Abstract: Fusion breeding blanket as one of the key in-vessel components performs the functions of breeding the tritium, removing the nuclear heat and heat flux from plasma chamber as well as acting as part of shielding system. The radial build design which determines the arrangement of function zones and material properties on the radial direction is the basis of the detailed design of fusion breeding blanket. For facilitating the radial build design, this study aims for developing a pre-design code to optimize the radial build of blanket with considering the performance of nuclear and thermal-hydraulic simultaneously. Two main features of this code are: (1) Coupling of the neutronics analysis with the thermal-hydraulic analysis to speed up the analysis progress; (2) preliminary optimization algorithm using one or multiple specified objectives subject to the design criteria in the form of constrains imposed on design variables and performance parameters within the possible engineering ranges. This pre-design code has been applied to the conceptual design of water-cooled ceramic breeding blanket in project of China fusion engineering testing reactor (CFETR).

  2. Nuclear-thermal-coupled optimization code for the fusion breeding blanket conceptual design

    Energy Technology Data Exchange (ETDEWEB)

    Li, Jia, E-mail: lijia@ustc.edu.cn [School of Nuclear Science and Technology, University of Science and Technology of China, Hefei 230027, Anhui (China); Jiang, Kecheng; Zhang, Xiaokang [Institute of Plasma Physics, Chinese Academy of Sciences, Hefei 230031, Anhui (China); Nie, Xingchen [School of Nuclear Science and Technology, University of Science and Technology of China, Hefei 230027, Anhui (China); Zhu, Qinjun; Liu, Songlin [Institute of Plasma Physics, Chinese Academy of Sciences, Hefei 230031, Anhui (China)

    2016-12-15

    Highlights: • A nuclear-thermal-coupled predesign code has been developed for optimizing the radial build arrangement of fusion breeding blanket. • Coupling module aims at speeding up the efficiency of design progress by coupling the neutronics calculation code with the thermal-hydraulic analysis code. • Radial build optimization algorithm aims at optimal arrangement of breeding blanket considering one or multiple specified objectives subject to the design criteria such as material temperature limit and available TBR. - Abstract: Fusion breeding blanket as one of the key in-vessel components performs the functions of breeding the tritium, removing the nuclear heat and heat flux from plasma chamber as well as acting as part of shielding system. The radial build design which determines the arrangement of function zones and material properties on the radial direction is the basis of the detailed design of fusion breeding blanket. For facilitating the radial build design, this study aims for developing a pre-design code to optimize the radial build of blanket with considering the performance of nuclear and thermal-hydraulic simultaneously. Two main features of this code are: (1) Coupling of the neutronics analysis with the thermal-hydraulic analysis to speed up the analysis progress; (2) preliminary optimization algorithm using one or multiple specified objectives subject to the design criteria in the form of constrains imposed on design variables and performance parameters within the possible engineering ranges. This pre-design code has been applied to the conceptual design of water-cooled ceramic breeding blanket in project of China fusion engineering testing reactor (CFETR).

  3. Production of Hev b5 as a fluorescent biotin-binding tripartite fusion protein in insect cells

    International Nuclear Information System (INIS)

    Nordlund, Henri R.; Laitinen, Olli H.; Uotila, Sanna T.H.; Kulmala, Minna; Kalkkinen, Nisse; Kulomaa, Markku S.

    2005-01-01

    The presented green fluorescent protein and streptavidin core-based tripartite fusion system provides a simple and efficient way for the production of proteins fused to it in insect cells. This fusion protein forms a unique tag, which serves as a multipurpose device enabling easy optimization of production, one-step purification via streptavidin-biotin interaction, and visualization of the fusion protein during downstream processing and in applications. In the present study, we demonstrate the successful production, purification, and detection of a natural rubber latex allergen Hev b5 with this system. We also describe the production of another NRL allergen with the system, Hev b1, which formed large aggregates and gave small yields in purification. The aggregates were detected at early steps by microscopical inspection of the infected insect cells producing this protein. Therefore, this fusion system can also be utilized as a fast indicator of the solubility of the expressed fusion proteins and may therefore be extremely useful in high-throughput expression approaches

  4. Production of Hev b5 as a fluorescent biotin-binding tripartite fusion protein in insect cells.

    Science.gov (United States)

    Nordlund, Henri R; Laitinen, Olli H; Uotila, Sanna T H; Kulmala, Minna; Kalkkinen, Nisse; Kulomaa, Markku S

    2005-10-14

    The presented green fluorescent protein and streptavidin core-based tripartite fusion system provides a simple and efficient way for the production of proteins fused to it in insect cells. This fusion protein forms a unique tag, which serves as a multipurpose device enabling easy optimization of production, one-step purification via streptavidin-biotin interaction, and visualization of the fusion protein during downstream processing and in applications. In the present study, we demonstrate the successful production, purification, and detection of a natural rubber latex allergen Hev b5 with this system. We also describe the production of another NRL allergen with the system, Hev b1, which formed large aggregates and gave small yields in purification. The aggregates were detected at early steps by microscopical inspection of the infected insect cells producing this protein. Therefore, this fusion system can also be utilized as a fast indicator of the solubility of the expressed fusion proteins and may therefore be extremely useful in high-throughput expression approaches.

  5. Junction region of EWS-FLI1 fusion protein has a dominant negative effect in Ewing's sarcoma in vitro.

    Science.gov (United States)

    Jully, Babu; Vijayalakshmi, Ramshankar; Gopal, Gopisetty; Sabitha, Kesavan; Rajkumar, Thangarajan

    2012-11-12

    Ewing's sarcoma is a malignancy characterized by a specific 11:22 chromosomal translocation which generates a novel EWS-FLI1 fusion protein functioning as an aberrant transcription factor. In the present study, we have further characterized the junction region of the EWS-FLI1 fusion protein. In-silico model of EWS-FLI1 fusion protein was analysed for ligand binding sites, and a putative region (amino acid (aa) 251-343 of the type 1 fusion protein) in the vicinity of the fusion junction was cloned and expressed using bacterial expression. The recombinant protein was characterized by Circular Dichroism (CD). We then expressed aa 251-280 ectopically in Ewing's sarcoma cell-line and its effect on cell proliferation, tumorigenicity and expression of EWS-FLI1 target genes were analysed. Our modelling analysis indicated that Junction region (aa 251-343) encompasses potential ligand biding sites in the EWS-FLI1 protein and when expressed in bacteria was present as soluble form. Ectopically expressing this region in Ewing's sarcoma cells inhibited tumorigenicity, and EWS-FLI1 target genes indicating a dominant negative biological effect. Junction region can be exploited further as target for drug development in future to specifically target EWS-FLI1 in Ewing's Sarcoma.

  6. Self-Assembly of Spider Silk-Fusion Proteins Comprising Enzymatic and Fluorescence Activity.

    Science.gov (United States)

    Humenik, Martin; Mohrand, Madeleine; Scheibel, Thomas

    2018-04-18

    The recombinant spider silk protein eADF4(C16) was genetically fused either with esterase 2 (EST2) or green fluorescent protein (GFP). The fusions EST-eADF4(C16) and GFP-eADF4(C16) were spectroscopically investigated and showed native structures of EST and GFP. The structural integrity was confirmed by the enzymatic activity of EST and the fluorescence of GFP. The spider silk moiety retained its intrinsically unstructured conformation in solution and the self-assembly into either nanofibrils or nanoparticles could be controlled by the concentration of phosphate. Particles, however, showed significantly lower activity of the EST and GFP domains likely caused by a steric hindrance. However, upon self-assembly of EST-eADF4(C16) and GFP-eADF4(C16) into fibrils the protein activities were retained. In general, the fusion of globular enzymes with the spider silk domain allows the generation of fibrous biomaterials with catalytic or light emitting properties.

  7. Delayed Toxicity Associated with Soluble Anthrax Toxin Receptor Decoy-Ig Fusion Protein Treatment

    Science.gov (United States)

    Cote, Christopher; Welkos, Susan; Manchester, Marianne; Young, John A. T.

    2012-01-01

    Soluble receptor decoy inhibitors, including receptor-immunogloubulin (Ig) fusion proteins, have shown promise as candidate anthrax toxin therapeutics. These agents act by binding to the receptor-interaction site on the protective antigen (PA) toxin subunit, thereby blocking toxin binding to cell surface receptors. Here we have made the surprising observation that co-administration of receptor decoy-Ig fusion proteins significantly delayed, but did not protect, rats challenged with anthrax lethal toxin. The delayed toxicity was associated with the in vivo assembly of a long-lived complex comprised of anthrax lethal toxin and the receptor decoy-Ig inhibitor. Intoxication in this system presumably results from the slow dissociation of the toxin complex from the inhibitor following their prolonged circulation. We conclude that while receptor decoy-Ig proteins represent promising candidates for the early treatment of B. anthracis infection, they may not be suitable for therapeutic use at later stages when fatal levels of toxin have already accumulated in the bloodstream. PMID:22511955

  8. HUWE1 and TRIP12 Collaborate in Degradation of Ubiquitin-Fusion Proteins and Misframed Ubiquitin

    DEFF Research Database (Denmark)

    Poulsen, Esben G; Steinhauer, Cornelia; Lees, Michael

    2012-01-01

    In eukaryotic cells an uncleavable ubiquitin moiety conjugated to the N-terminus of a protein signals the degradation of the fusion protein via the proteasome-dependent ubiquitin fusion degradation (UFD) pathway. In yeast the molecular mechanism of the UFD pathway has been well characterized...... in degradation of the UFD substrate Ub(G76V)-YFP. The most significant hits from the screen were the E3 ubiquitin-protein ligase HUWE1, as well as PSMD7 and PSMD14 that encode proteasome subunits. Accordingly, knock down of HUWE1 led to an increase in the steady state level and a retarded degradation of the UFD...... substrate. Knock down of HUWE1 also led to a stabilization of the physiological UFD substrate UBB(+1). Precipitation experiments revealed that HUWE1 is associated with both the Ub(G76V)-YFP substrate and the 26S proteasome, indicating that it functions late in the UFD pathway. Double knock down of HUWE1...

  9. F-18 Labeled Diabody-Luciferase Fusion Proteins for Optical-ImmunoPET

    Energy Technology Data Exchange (ETDEWEB)

    Wu, Anna M. [Univ. of California, Los Angeles, CA (United States)

    2013-01-18

    The goal of the proposed work is to develop novel dual-labeled molecular imaging probes for multimodality imaging. Based on small, engineered antibodies called diabodies, these probes will be radioactively tagged with Fluorine-18 for PET imaging, and fused to luciferases for optical (bioluminescence) detection. Performance will be evaluated and validated using a prototype integrated optical-PET imaging system, OPET. Multimodality probes for optical-PET imaging will be based on diabodies that are dually labeled with 18F for PET detection and fused to luciferases for optical imaging. 1) Two sets of fusion proteins will be built, targeting the cell surface markers CEA or HER2. Coelenterazine-based luciferases and variant forms will be evaluated in combination with native substrate and analogs, in order to obtain two distinct probes recognizing different targets with different spectral signatures. 2) Diabody-luciferase fusion proteins will be labeled with 18F using amine reactive [18F]-SFB produced using a novel microwave-assisted, one-pot method. 3) Sitespecific, chemoselective radiolabeling methods will be devised, to reduce the chance that radiolabeling will inactivate either the target-binding properties or the bioluminescence properties of the diabody-luciferase fusion proteins. 4) Combined optical and PET imaging of these dual modality probes will be evaluated and validated in vitro and in vivo using a prototype integrated optical-PET imaging system, OPET. Each imaging modality has its strengths and weaknesses. Development and use of dual modality probes allows optical imaging to benefit from the localization and quantitation offered by the PET mode, and enhances the PET imaging by enabling simultaneous detection of more than one probe.

  10. Engineering of a Potent Recombinant Lectin-Toxin Fusion Protein to Eliminate Human Pluripotent Stem Cells.

    Science.gov (United States)

    Tateno, Hiroaki; Saito, Sayoko

    2017-07-10

    The use of human pluripotent stem cells (hPSCs) such as human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs) in regenerative medicine is hindered by their tumorigenic potential. Previously, we developed a recombinant lectin-toxin fusion protein of the hPSC-specific lectin rBC2LCN, which has a 23 kDa catalytic domain (domain III) of Pseudomonas aeruginosa exotoxin A (rBC2LCN-PE23). This fusion protein could selectively eliminate hPSCs following its addition to the cell culture medium. Here we conjugated rBC2LCN lectin with a 38 kDa domain of exotoxin A containing domains Ib and II in addition to domain III (PE38). The developed rBC2LCN-PE38 fusion protein could eliminate 50% of 201B7 hPSCs at a concentration of 0.003 μg/mL (24 h incubation), representing an approximately 556-fold higher activity than rBC2LCN-PE23. Little or no effect on human fibroblasts, human mesenchymal stem cells, and hiPSC-derived hepatocytes was observed at concentrations lower than 1 μg/mL. Finally, we demonstrate that rBC2LCN-PE38 selectively eliminates hiPSCs from a mixed culture of hiPSCs and hiPSC-derived hepatocytes. Since rBC2LCN-PE38 can be prepared from soluble fractions of E. coli culture at a yield of 9 mg/L, rBC2LCN-PE38 represents a practical reagent to remove human pluripotent stem cells residing in cultured cells destined for transplantation.

  11. S-layer fusion protein as a tool functionalizing emulsomes and CurcuEmulsomes for antibody binding and targeting.

    Science.gov (United States)

    Ucisik, Mehmet H; Küpcü, Seta; Breitwieser, Andreas; Gelbmann, Nicola; Schuster, Bernhard; Sleytr, Uwe B

    2015-04-01

    Selective targeting of tumor cells by nanoparticle-based drug delivery systems is highly desirable because it maximizes the drug concentration at the desired target while simultaneously protecting the surrounding healthy tissues. Here, we show a design for smart nanocarriers based on a biomimetic approach that utilizes the building principle of virus envelope structures. Emulsomes and CurcuEmulsomes comprising a tripalmitin solid core surrounded by phospholipid layers are modified by S-layer proteins that self-assemble into a two-dimensional array to form a surface layer. One significant advantage of this nanoformulation is that it increases the solubility of the lipophilic anti-cancer agent curcumin in the CurcuEmulsomes by a factor of 2700. In order to make the emulsomes specific for IgG, the S-layer protein is fused with two protein G domains. This S-layer fusion protein preserves its recrystallization characteristics, forming an ordered surface layer (square lattice with 13 nm unit-by-unit distance). The GG domains are presented in a predicted orientation and exhibit a selective binding affinity for IgG. Copyright © 2015 The Authors. Published by Elsevier B.V. All rights reserved.

  12. First wall studies of a laser-fusion hybrid reactor design

    International Nuclear Information System (INIS)

    Hovingh, J.

    1976-09-01

    The design of a first wall for a 20 MW thermonuclear power laser fusion hybrid reactor is presented. The 20 mm thick graphite first wall is located 3.5 m from the DT microexplosion with a thermonuclear yield of 10 MJ. Estimates of the energy deposition, temperature, stresses, and material vaporized from the first wall due to the interaction of the x-rays, charged particle debris, and reflected laser light with the graphite are presented, along with a brief description of the analytical methods used for these estimations. Graphite is a viable first wall material for inertially-confined fusion reactors, with lifetimes of a year possible

  13. Identifying heavy-ion-beam fusion design and system features with high economic leverage

    International Nuclear Information System (INIS)

    Meier, W.R.; Hogan, W.J.

    1985-01-01

    We have conducted parametric economic studies for heavy-ion-beam fusion electric power plants. We examined the effects on the cost of electricity of several design parameters: maximum achievable chamber pulse rate, driver cost, target gain, and electric conversion efficiency, and net electric power. We found with reasonable assumptions on driver cost, target gain, and electric conversion efficiency, a 2 to 3 GWe heavy-ion-beam fusion power plant, with a chamber pulse rate of 5 to 10 Hz, can be competitive with nuclear and coal power plants

  14. Design study of a fusion-driven tokamak hybrid reactor for fissile fuel production. Final report

    International Nuclear Information System (INIS)

    Rose, R.P.

    1979-05-01

    This study evaluated conceptual approaches for a tokamak fusion-driven fuel producing reactor. The conceptual design of this hybrid reactor was based on using projected state-of-the-art technology for the late 1980s. This reactor would be a demonstration plant and, therefore, first-of-a-kind considerations have been included. The conceptual definitions of two alternatives for the fusion driver were evaluated. A Two-Component Tokamak (TCT) concept, based on the TFTR plasma physics parameters, was compared to a Beam-Driven Thermonuclear (BDTN) concept, based on the USSR T-20 plasma physics parameters

  15. The heat transport system and plant design for the HYLIFE-2 fusion reactor

    International Nuclear Information System (INIS)

    Hoffman, M.A.

    1990-01-01

    HYLIFE is the name given to a family of self-healing liquid-wall reactor concepts for inertial confinement fusion. This HYLIFE-II concept employs the molten salt, Flibe, for the liquid jets instead of liquid lithium used in the original HYLIFE-I study. A preliminary conceptual design study of the heat transport system and the balance of plant of the HYLIFE-II fusion power plant is described in this paper with special emphasis on a scoping study to determine the best intermediate heat exchanger geometry and flow conditions for minimum cost of electricity. 11 refs., 8 figs

  16. Laser fusion reactor design in a fast ignition with a dry wall chamber

    International Nuclear Information System (INIS)

    Ogawa, Yichi; Goto, Takuya; Ninomiya, Daisuke; Hiwatari, Ryoji; Asaoka, Yoshiyuki; Okano, Kunihiko

    2007-01-01

    One of the critical issues in laser fusion reactor design is high pulse heat load on the first wall by the X-rays and the fast/debris ions from fusion burn. There are mainly two concepts for the first wall of laser fusion reactor, a dry wall and a liquid metal wall. We should notice that the fast ignition method can achieve sufficiently high pellet gain with smaller (about 1/10 of the conventional central ignition method) input energy. To take advantage of this property, the design of a laser fusion reactor with a small size dry wall chamber may become possible. Since a small fusion pulse leads to a small electric power, high repetition of laser irradiation is required to keep sufficient electric power. Then we tried to design a laser fusion reactor with a dry wall chamber and a high repetition laser. This is a new challenging path to realize a laser fusion plant. Based on the point model of the core plasma, we have estimated that fusion energy in one pulse can be reduced to be 40 MJ with a pellet gain around G>100. To evaluate the validity of this simple estimation and to optimize the pellet design and the pulse shaping for the fast ignition scenario, we have introduced 1-D hydrodynamic simulation code ILESTA-1D and carried out implosion simulations. Since the code is one-dimensional, the detailed physics process of fast heating cannot be reproduced. Thus the fast heating is reflected in the code as the additional artificial heating source in the energy equation. It is modeled as a homogeneous heating of electrons in core region at the time just before when the maximum compression is achieved. At present we obtained the pellet gain G∝100 with the same input energy as the above estimation by a simple point model (350kJ for implosion, 50kJ for heating and assuming 20% coupling of heating laser). A dry wall is exposed to several threats due to the cyclic load by the high energy X-ray and charged particles: surface melting, physical and chemical sputtering

  17. Design of a high-flux test assembly for the Fusion Materials Irradiation Test Facility

    International Nuclear Information System (INIS)

    Opperman, E.K.; Vogel, M.A.

    1982-01-01

    The Fusion Material Test Facility (FMIT) will provide a high flux fusion-like neutron environment in which a variety of structural and non-structural materials irradiations can be conducted. The FMIT experiments, called test assemblies, that are subjected to the highest neutron flux magnitudes and associated heating rates will require forced convection liquid metal cooling systems to remove the neutron deposited power and maintain test specimens at uniform temperatures. A brief description of the FMIT facility and experimental areas is given with emphasis on the design, capabilities and handling of the high flux test assembly

  18. Design and Implementation of Multi Agentbased Information Fusion System for Decision Making Support (A Case Study on Military Operation

    Directory of Open Access Journals (Sweden)

    Arwin Datunaya Wahyudi Sumari

    2013-09-01

    Full Text Available Quick, accurate, and complete information is highly required for supporting strategically impact decision making in a Military Operation (MO in order to reduce the decision cycle and to minimize the loss. For that purpose, we propose, design and implement a hierarchical Multi Agentbased Information Fusion System for Decision Making Support (MAIFSDMS. The information fusion is implemented by applying Maximum Score of the Total Sum of Joint Probabilities (MSJP fusion method and is done by a collection of Information Fusion Agents (IFA that forms a multiagent system. MAIFS uses a combination of generalization of Dasarathy and Joint Director’s Laboratory (JDL process models for information fusion mechanism. Information fusion products that are displayed in graphical forms provide comprehensive information regarding the MO’s area dynamics. By observing the graphics resulted from the information fusion, the commandant will have situational awareness and knowledge in order to make the most accurate strategic de cision as fast as possible.

  19. Achievements and Challenges in Computational Protein Design.

    Science.gov (United States)

    Samish, Ilan

    2017-01-01

    Computational protein design (CPD), a yet evolving field, includes computer-aided engineering for partial or full de novo designs of proteins of interest. Designs are defined by a requested structure, function, or working environment. This chapter describes the birth and maturation of the field by presenting 101 CPD examples in a chronological order emphasizing achievements and pending challenges. Integrating these aspects presents the plethora of CPD approaches with the hope of providing a "CPD 101". These reflect on the broader structural bioinformatics and computational biophysics field and include: (1) integration of knowledge-based and energy-based methods, (2) hierarchical designated approach towards local, regional, and global motifs and the integration of high- and low-resolution design schemes that fit each such region, (3) systematic differential approaches towards different protein regions, (4) identification of key hot-spot residues and the relative effect of remote regions, (5) assessment of shape-complementarity, electrostatics and solvation effects, (6) integration of thermal plasticity and functional dynamics, (7) negative design, (8) systematic integration of experimental approaches, (9) objective cross-assessment of methods, and (10) successful ranking of potential designs. Future challenges also include dissemination of CPD software to the general use of life-sciences researchers and the emphasis of success within an in vivo milieu. CPD increases our understanding of protein structure and function and the relationships between the two along with the application of such know-how for the benefit of mankind. Applied aspects range from biological drugs, via healthier and tastier food products to nanotechnology and environmentally friendly enzymes replacing toxic chemicals utilized in the industry.

  20. Reduction in Brain Heparan Sulfate with Systemic Administration of an IgG Trojan Horse-Sulfamidase Fusion Protein in the Mucopolysaccharidosis Type IIIA Mouse.

    Science.gov (United States)

    Boado, Ruben J; Lu, Jeff Zhiqiang; Hui, Eric Ka-Wai; Pardridge, William M

    2018-02-05

    Mucopolysaccharidosis Type IIIA (MPSIIIA), also known as Sanfilippo A syndrome, is an inherited neurodegenerative disease caused by mutations in the lysosomal enzyme, N-sulfoglucosamine sulfohydrolase (SGSH), also known as sulfamidase. Mutations in the SGSH enzyme, the only mammalian heparan N-sulfatase, cause accumulation of lysosomal inclusion bodies in brain cells comprising heparan sulfate (HS) glycosaminoglycans (GAGs). Treatment of MPSIIIA with intravenous recombinant SGSH is not possible because this large molecule does not cross the blood-brain barrier (BBB). BBB penetration by SGSH was enabled in the present study by re-engineering this enzyme as an IgG-SGSH fusion protein, where the IgG domain is a chimeric monoclonal antibody (mAb) against the mouse transferrin receptor (TfR), designated the cTfRMAb. The IgG domain of the fusion protein acts as a molecular Trojan horse to deliver the enzyme into brain via transport on the endogenous BBB TfR. The cTfRMAb-SGSH fusion protein bound to the mouse TfR with high affinity, ED 50 = 0.74 ± 0.07 nM, and retained high SGSH enzyme activity, 10 043 ± 1003 units/mg protein, which is comparable to recombinant human SGSH. Male and female MPSIIIA mice, null for the SGSH enzyme, were treated for 6 weeks with thrice-weekly intraperitoneal injections of vehicle, 5 mg/kg of the cTfRMAb alone, or 5 mg/kg of the cTfRMAb-SGSH fusion protein, starting at the age of 2 weeks, and were euthanized 1 week after the last injection. Brain and liver HS, as determined by liquid chromatography-mass spectrometry, were elevated 30-fold and 36-fold, respectively, in the MPSIIIA mouse. Treatment of the mice with the cTfRMAb-SGSH fusion protein caused a 70% and 85% reduction in brain and liver HS, respectively. The reduction in brain HS was associated with a 28% increase in latency on the rotarod test of motor activity in male mice. The mice exhibited no injection related reactions, and only a low titer end of study antidrug antibody

  1. Neutron shield analysis and design for the PDX fusion facility

    International Nuclear Information System (INIS)

    Grimesey, R.A.; Nigg, D.W.; Scott, A.J.; Wheeler, F.J.; Jassby, D.L.; Perry, E.D.

    1979-01-01

    The basic component of the biological shield for PDX is an existing 81 cm thick high-density concrete shielding wall surrounding the machine. The principal additional shielding requirement is a roof shield over the machine to reduce air-scattered skyshine dose into the PDX control room and to the site boundary. The roof shield is designed in removable sections on a steel support structure permitting overhead crane access to major PDX components. After analysis of a number of alternate concepts, a roof shield consisting of 50 cm of water in polyethylene tanks was selected to meet design objectives of effectiveness, weight, removability, and cost

  2. Conceptual design study of fusion experimental reactor (FY 86 FER)

    International Nuclear Information System (INIS)

    Kobayashi, Takeshi; Yamada, Masao; Mizoguchi, Tadanori

    1987-09-01

    This report describes the results of the investigation on critical issues of FY 86 FER reactor configuration/structure design. Accuracy evaluation of shielding calculation and crack growth prediction of first wall and divertor based on the elastic-plastic fracture mechanics were performed. Further, optimization of shield configuration, graphite first wall armor and flexifility of reactor were investigated to support future design work. Feasibilities of innovative ideas were also examined, such as the ripple insert effect and the application of shape memory alloys. (author)

  3. Safety considerations in the design of the Fusion Engineering Device

    International Nuclear Information System (INIS)

    Barrett, R.J.

    1983-01-01

    The US Department of Energy (DOE) regulations and guidelines for radiation protection have been reviewed and are being applied to the device design. Direct radiation protection is provided by the device shield and the reactor building walls. Radiation from the activated device components and the tritium fuel is to be controlled with shielding, contamination control, and ventilation. The potential release of tritium from the plant has influenced the selection of reactor building and plant designs and specifications. The safety of the plant workers is affected primarily by the radiation from the activated device components and from plasma chamber debris

  4. Conceptual design of a fusion-fission hybrid reactor for transmutation of high level nuclear waste

    International Nuclear Information System (INIS)

    Qiu, L.J.; Wu, Y.C.; Yang, Y.W.; Wu, Y.; Luan, G.S.; Xu, Q.; Guo, Z.J.; Xiao, B.J.

    1994-01-01

    To assess the feasibility of the transmutation of long-lived radioactive waste using fusion-fission hybrid reactors, we are studying all the possible types of blanket, including a comparison of the thermal and fast neutron spectrum blankets. Conceptual designs of a small tokamak hybrid blanket with small inventory of actinides and fission products are presented. The small inventory of wastes makes the system safer. The small hybrid reactor system based on a fusion core with experimental parameters to be realized in the near future can effectively transmute actinides and fission products at a neutron wall loading of 1MWm -2 . An innovative energy system is also presented, including a fusion driver, fuel breeder, high level waste transmuter, fission reactor and so on. An optimal combination of all types of reactor is proposed in the system. ((orig.))

  5. Conceptual design of tritium production fusion reactor based on spherical torus

    International Nuclear Information System (INIS)

    He Kaihui; Huang Jinhua

    2003-01-01

    Conceptual design of an advanced tritium production fusion reactor based on spherical torus, which is intermediate application of fusion energy, was presented in this paper. Differing from the traditional tokamak tritium production reactor design, advanced plasma physics performance and compact structural characteristics of ST were used to minimize tritium leakage and maximize tritium breeding ratio with arrangement of tritium production blankets within vacuum vessel as possible in order to produce 1 kg excess tritium except need of self-sufficient plasma core with 40% or more corresponding plant availability. Based on 2D neutronics calculation, preliminary conceptual design of ST-TPR was presented, providing the backgrounds and reference for next detailed conceptual design

  6. Design of an 18 Tesla, tandem mirror, fusion reactor, hybrid choke coil

    International Nuclear Information System (INIS)

    Parmer, J.F.; Agarwal, K.; Gurol, H.; Mancuso, A.; Michels, P.H.; Peck, S.D.; Burgeson, J.; Dalder, E.N.

    1987-01-01

    A hybrid, part normal part superconducting 18-Tesla solenoid choke coil is designed for a tandem mirror fusion reactor. The present state of the art is represented by the 12-Tesla, superconducting NbSn coil. Future applications other than tandem mirror fusion devices needing high field solenoids might require hybrid magnets of the type described herein. The hybrid design was generated because of critical field performance limitations on present, practical superconducting wires. A hybrid design might be required (due to structural limits) even if the critical field were higher. Also, hybrids could be a cost-effective way of getting very high fields for certain applications. The 18-Tesla solenoid described is composed of an inner coil made of water-cooled, high-strength zirconium copper which generates 3 Tesla. A superconducting NbSn background coil contributes the remaining 15 Tesla. The focus of the design study was on the inner coil. Demonstration fabrication and testing was performed

  7. A mature and fusogenic form of the Nipah virus fusion protein requires proteolytic processing by cathepsin L

    International Nuclear Information System (INIS)

    Pager, Cara Theresia; Craft, Willie Warren; Patch, Jared; Dutch, Rebecca Ellis

    2006-01-01

    The Nipah virus fusion (F) protein is proteolytically processed to F 1 + F 2 subunits. We demonstrate here that cathepsin L is involved in this important maturation event. Cathepsin inhibitors ablated cleavage of Nipah F. Proteolytic processing of Nipah F and fusion activity was dramatically reduced in cathepsin L shRNA-expressing Vero cells. Additionally, Nipah virus F-mediated fusion was inhibited in cathepsin L-deficient cells, but coexpression of cathepsin L restored fusion activity. Both purified cathepsin L and B could cleave immunopurified Nipah F protein, but only cathepsin L produced products of the correct size. Our results suggest that endosomal cathepsins can cleave Nipah F, but that cathepsin L specifically converts Nipah F to a mature and fusogenic form

  8. Systems approach for condition management design: JET neutral beam system-A fusion case study

    Energy Technology Data Exchange (ETDEWEB)

    Khella, M., E-mail: M.Khella@lboro.ac.uk [Systems Engineering Innovation Centre (SEIC) - BAE Systems, Loughborough University, Holywell Park, Leicestershire LE11 3TU (United Kingdom); Pearson, J. [Systems Engineering Innovation Centre (SEIC) - BAE Systems, Loughborough University, Holywell Park, Leicestershire LE11 3TU (United Kingdom); Dixon, R. [Electronic and Electrical Engineering, Loughborough University, Leicestershire LE11 3TU (United Kingdom); Ciric, D.; Day, I.; King, R.; Milnes, J.; Stafford-Allen, R. [EURATOM/CCFE Fusion Association, Culham Science Centre, Oxfordshire OX14 3DB (United Kingdom)

    2011-10-15

    The maturation of any new technology can be coarsely divided into three stages of a development lifecycle: (1) fundamental research, (2) experimental rig development and testing through to (3) commercialization. With the enhancement of machines like JET, the building of ITER and the initiation of DEMO design activities, the fusion community is moving from stages 1 and 2 towards stage 3. One of the consequences of this transition will be a shift in emphasis from scientific achievement to maximizing machine reliability and availability. The fusion community should therefore be preparing itself for this shift by examining all methods and tools utilized in established engineering sectors that might help to improve these fundamental performance parameters. To this end, the Culham Centre for Fusion Energy (CCFE) has proactively engaged with UK industry to examine whether the development of condition management (CM) systems could help improve such performance parameters. This paper describes an initial CM design case study on the JET neutral beam system. The primary output of this study was the development of a CM design methodology that captures existing experience in fault detection, and classification as well as new methods for fault diagnosis. A summary of the methods used and the potential benefits of data fusion are presented here.

  9. Mechanical design for a large fusion laser system

    International Nuclear Information System (INIS)

    Hurley, C.A.

    1979-01-01

    The Nova Mechanical Systems Group at LLL is responsible for the design, fabrication, and installation of all laser chain components, for the stable support structure that holds them, and for the beam lines that transport the laser beam to the target system. This paper is an overview of the group's engineering effort, emphasizing new developments

  10. Design of the 1-Mw, 200-Ghz, Fom Fusion Fem

    NARCIS (Netherlands)

    Urbanus, W. H.; Best, R. W. B.; Bongers, W. A.; Vaningen, A. M.; Manintveld, P.; Sterk, A. B.; Verhoeven, A. G. A.; van der Wiel, M. J.; Caplan, M.; Bratman, V. L.; Denisov, G. G.; Varfolomeev, A. A.; Khlebnikov, A. S.

    1993-01-01

    The FOM Institute for Plasma Physics has obtained funding for the development of a 1 MW, long pulse, 140-250 GHz free-electron maser. The engineering design is presently being performed in an international collaboration. In this paper the main components of the free-electron maser, the electron beam

  11. An Electrostatic Free-Electron Maser for Fusion - Design Considerations

    NARCIS (Netherlands)

    van Amersfoort, P. W.; Urbanus, W. H.; Verhoeven, A. G. A.; Verheul, A.; Sterk, A. B.; Vaningen, A. M.; van der Wiel, M. J.

    1991-01-01

    For the next generation of large tokamaks, efficient mm-wave sources at frequencies of up to 300 GHz and unit size of 1 MW cw will be required. The design of a free electron maser for this application, based on a dc electrostatic accelerator, is discussed.

  12. HUWE1 and TRIP12 collaborate in degradation of ubiquitin-fusion proteins and misframed ubiquitin.

    Directory of Open Access Journals (Sweden)

    Esben G Poulsen

    Full Text Available In eukaryotic cells an uncleavable ubiquitin moiety conjugated to the N-terminus of a protein signals the degradation of the fusion protein via the proteasome-dependent ubiquitin fusion degradation (UFD pathway. In yeast the molecular mechanism of the UFD pathway has been well characterized. Recently the human E3 ubiquitin-protein ligase TRIP12 was connected with the UFD pathway, but little is otherwise known about this system in mammalian cells. In the present work, we utilized high-throughput imaging on cells transfected with a targeted siRNA library to identify components involved in degradation of the UFD substrate Ub(G76V-YFP. The most significant hits from the screen were the E3 ubiquitin-protein ligase HUWE1, as well as PSMD7 and PSMD14 that encode proteasome subunits. Accordingly, knock down of HUWE1 led to an increase in the steady state level and a retarded degradation of the UFD substrate. Knock down of HUWE1 also led to a stabilization of the physiological UFD substrate UBB(+1. Precipitation experiments revealed that HUWE1 is associated with both the Ub(G76V-YFP substrate and the 26S proteasome, indicating that it functions late in the UFD pathway. Double knock down of HUWE1 and TRIP12 resulted in an additive stabilization of the substrate, suggesting that HUWE1 and TRIP12 function in parallel during UFD. However, even when both HUWE1 and TRIP12 are downregulated, ubiquitylation of the UFD substrate was still apparent, revealing functional redundancy between HUWE1, TRIP12 and yet other ubiquitin-protein ligases.

  13. New design of cable-in-conduit conductor for application in future fusion reactors

    Science.gov (United States)

    Qin, Jinggang; Wu, Yu; Li, Jiangang; Liu, Fang; Dai, Chao; Shi, Yi; Liu, Huajun; Mao, Zhehua; Nijhuis, Arend; Zhou, Chao; Yagotintsev, Konstantin A.; Lubkemann, Ruben; Anvar, V. A.; Devred, Arnaud

    2017-11-01

    The China Fusion Engineering Test Reactor (CFETR) is a new tokamak device whose magnet system includes toroidal field, central solenoid (CS) and poloidal field coils. The main goal is to build a fusion engineering tokamak reactor with about 1 GW fusion power and self-sufficiency by blanket. In order to reach this high performance, the magnet field target is 15 T. However, the huge electromagnetic load caused by high field and current is a threat for conductor degradation under cycling. The conductor with a short-twist-pitch (STP) design has large stiffness, which enables a significant performance improvement in view of load and thermal cycling. But the conductor with STP design has a remarkable disadvantage: it can easily cause severe strand indentation during cabling. The indentation can reduce the strand performance, especially under high load cycling. In order to overcome this disadvantage, a new design is proposed. The main characteristic of this new design is an updated layout in the triplet. The triplet is made of two Nb3Sn strands and one soft copper strand. The twist pitch of the two Nb3Sn strands is large and cabled first. The copper strand is then wound around the two superconducting strands (CWS) with a shorter twist pitch. The following cable stages layout and twist pitches are similar to the ITER CS conductor with STP design. One short conductor sample with a similar scale to the ITER CS was manufactured and tested with the Twente Cable Press to investigate the mechanical properties, AC loss and internal inspection by destructive examination. The results are compared to the STP conductor (ITER CS and CFETR CSMC) tests. The results show that the new conductor design has similar stiffness, but much lower strand indentation than the STP design. The new design shows potential for application in future fusion reactors.

  14. Muon-catalyzed fusion experiment target and detector system. Preliminary design report

    International Nuclear Information System (INIS)

    Jones, S.E.; Watts, K.D.; Caffrey, A.J.; Walter, J.B.

    1982-03-01

    We present detailed plans for the target and particle detector systems for the muon-catalyzed fusion experiment. Requirements imposed on the target vessel by experimental conditions and safety considerations are delineated. Preliminary designs for the target vessel capsule and secondary containment vessel have been developed which meet these requirements. In addition, the particle detection system is outlined, including associated fast electronics and on-line data acquisition. Computer programs developed to study the target and detector system designs are described

  15. Structural characterization of respiratory syncytial virus fusion inhibitor escape mutants: homology model of the F protein and a syncytium formation assay

    International Nuclear Information System (INIS)

    Morton, Craig J.; Cameron, Rachel; Lawrence, Lynne J.; Lin Bo; Lowe, Melinda; Luttick, Angela; Mason, Anthony; McKimm-Breschkin, Jenny; Parker, Michael W.; Ryan, Jane; Smout, Michael; Sullivan, Jayne; Tucker, Simon P.; Young, Paul R.

    2003-01-01

    Respiratory syncytial virus (RSV) is a ubiquitous human pathogen and the leading cause of lower respiratory tract infections in infants. Infection of cells and subsequent formation of syncytia occur through membrane fusion mediated by the RSV fusion protein (RSV-F). A novel in vitro assay of recombinant RSV-F function has been devised and used to characterize a number of escape mutants for three known inhibitors of RSV-F that have been isolated. Homology modeling of the RSV-F structure has been carried out on the basis of a chimera derived from the crystal structures of the RSV-F core and a fragment from the orthologous fusion protein from Newcastle disease virus (NDV). The structure correlates well with the appearance of RSV-F in electron micrographs, and the residues identified as contributing to specific binding sites for several monoclonal antibodies are arranged in appropriate solvent-accessible clusters. The positions of the characterized resistance mutants in the model structure identify two promising regions for the design of fusion inhibitors

  16. Construction and use of a Cupriavidus necator H16 soluble hydrogenase promoter (PSH fusion to gfp (green fluorescent protein

    Directory of Open Access Journals (Sweden)

    Bat-Erdene Jugder

    2016-07-01

    Full Text Available Hydrogenases are metalloenzymes that reversibly catalyse the oxidation or production of molecular hydrogen (H2. Amongst a number of promising candidates for application in the oxidation of H2 is a soluble [Ni–Fe] uptake hydrogenase (SH produced by Cupriavidus necator H16. In the present study, molecular characterisation of the SH operon, responsible for functional SH synthesis, was investigated by developing a green fluorescent protein (GFP reporter system to characterise PSH promoter activity using several gene cloning approaches. A PSH promoter-gfp fusion was successfully constructed and inducible GFP expression driven by the PSH promoter under de-repressing conditions in heterotrophic growth media was demonstrated in the recombinant C. necator H16 cells. Here we report the first successful fluorescent reporter system to study PSH promoter activity in C. necator H16. The fusion construct allowed for the design of a simple screening assay to evaluate PSH activity. Furthermore, the constructed reporter system can serve as a model to develop a rapid fluorescent based reporter for subsequent small-scale process optimisation experiments for SH expression.

  17. Structural and kinetic analysis of the unnatural fusion protein 4-coumaroyl-CoA ligase::stilbene synthase

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Yechun; Yi, Hankuil; Wang, Melissa; Yu, Oliver; Jez, Joseph M. (WU); (Danforth)

    2012-10-24

    To increase the biochemical efficiency of biosynthetic systems, metabolic engineers have explored different approaches for organizing enzymes, including the generation of unnatural fusion proteins. Previous work aimed at improving the biosynthesis of resveratrol, a stilbene associated a range of health-promoting activities, in yeast used an unnatural engineered fusion protein of Arabidopsis thaliana (thale cress) 4-coumaroyl-CoA ligase (At4CL1) and Vitis vinifera (grape) stilbene synthase (VvSTS) to increase resveratrol levels 15-fold relative to yeast expressing the individual enzymes. Here we present the crystallographic and biochemical analysis of the 4CL::STS fusion protein. Determination of the X-ray crystal structure of 4CL::STS provides the first molecular view of an artificial didomain adenylation/ketosynthase fusion protein. Comparison of the steady-state kinetic properties of At4CL1, VvSTS, and 4CL::STS demonstrates that the fusion protein improves catalytic efficiency of either reaction less than 3-fold. Structural and kinetic analysis suggests that colocalization of the two enzyme active sites within 70 {angstrom} of each other provides the basis for enhanced in vivo synthesis of resveratrol.

  18. Collection of Summaries of reports on result of research at basic experiment device for nuclear fusion reactor blanket design, 1994

    International Nuclear Information System (INIS)

    1995-07-01

    The development of nuclear fusion reactors reached such stage that the generation of fusion power output comparable with the input power into core plasma is possible. At present, the engineering design of the international thermonuclear fusion experimental reactor, ITER, is advanced by the cooperation of Japan, USA, Europe and Russia, aiming at the start of operation at the beginning of 21st century. This meeting for reporting the results has been held every year, and this time, it was held on May 19, 1995 at University of Tokyo with the theme ''The interface properties of fusion reactor materials and the control of particle transport''. About 50 participants from academic, governmental and industrial circles discussed actively on the theme. Three lectures on the topics of fusion reactor engineering and materials and seven lectures on the basic experiment of fusion reactor blanket design related to the next period project were given at the meeting. (K.I.)

  19. Repair/maintenance design for tokamak experimental fusion reactor

    International Nuclear Information System (INIS)

    1978-10-01

    Repair and maintenance design for JXFR has been studied. The reactor is in eight modules so that a damaged module alone can be separated from the other modules and transferred from the reactor room to a repair shop. Design work covers overhaul procedure, dismounting equipments (overhead cranes, auto welder/cutter and remote handling equipments), transport system of a module (module mounting carriages and rotating carriage), repair equipment for blanket, earthquake-proof analysis of the reactor, reactor room structure, repair shop layout, management of radioactive wastes, time and the number of persons required for overhaul etc. Though the repair and maintenance system is almost complete, there still remain problems for further study in joints of blanket cooling piping, auto welder/cutter and earthquake-proof strength in reactor disassemblage. More detailed studies and R and D are necessary for engineering perfection. (author)

  20. Thermostructural design of the first wall/blanket for the TITAN-RFP fusion reactor

    International Nuclear Information System (INIS)

    Orient, G.E.; Blanchard, J.P.; Ghoniem, N.M.

    1987-01-01

    The mass power density, which is defined as the average power per unit mass within the magnet boundary, is a rough and general measure of economic competitiveness. Conn et al. (1985) have identified a target value of 100 kW(e)/tonne as a reasonable threshold for 'compact' commercial fusion systems. In pursuit of this goal, Hagenson et al. (1984) and Najmabadi et al. (1987) have pointed out the inherent characteristics of the RFP toroidal confinement concept which allow it to exceed this target value. It is inevitable that the compactness of the fusion power core will introduce a unique set of design issues. The special design concerns stem from high thermal surface fluxes, high bulk energy deposition by neutrons, and a relatively short blanket structural lifetime. In the TITAN-RFP, study Najmabadi et al. (1987) investigate a number of blanket (B) and first wall (FW) options suitable for high power density fusion reactors. Final choices were made for two designs: A high pressure aqueous blanket and a vanadium/lithium self-cooled blanket. The first design utilizes a pressurized aqueous loop containing a lithium compound dissolved in water, while the second design is based upon a self-cooled lithium-vanadium blanket. In this paper, we consider the beginning-of-life (BOL) thermostructural design and analysis of only the second concept. (orig./GL)

  1. Design of water detritiation system for fusion reactor

    International Nuclear Information System (INIS)

    Xie Bo; Wang Heyi; Liu Yunnu; Guan Rui

    2006-01-01

    The water detritiation system (WDS) of tritium plant for the International Thermonuclear Experimental Reactor (ITER) was designed. The concept of the Combined Electrolysis Catalytic Exchange and Gas Chromatography (CECE-GC) process was selected for the system and subsystems' descriptions of the WDS. ITER-WDS is characterised from the present demonstration system by rejecting the use of a recombiner and alkali electrolyzer, but a solid polymer electrolyzer (SPE) and a Pd/Ag membrane permeator system are adopted to recover tritium. (authors)

  2. Conceptual design Fusion Experimental Reactor (FER/ITER)

    International Nuclear Information System (INIS)

    Uehara, Kazuya; Nagashima, Takashi; Ikeda, Yoshitaka

    1991-11-01

    This report describes a conceptual design of Lower Hybrid Wave (LH) system for FER and ITER. In JAERI, the conceptual design of LH system for FER has been performed in these 3 years in parallel to that of ITER. There must be a common design part with ITER and FER. The physical requirement of LH system is the saving of volt · sec in the current start-up phase, and the current drive at the boundary region. The frequency of 5GHz is mainly chosen for avoidance of the α particle absorption and for the availability of electron tube development. Seventy-two klystrons (FER) and one hundred klystrons (ITER) are necessary to inject the 30 MW (FER) and 45-50 MW (ITER) rf power into plasma using 0.7 - 0.8 MW klystron per one tube. The launching system is the multi-junction type and the rf spectrum must be as sharp as possible with high directivity to improve the current drive efficiency. One port (FER) and two ports (ITER) are used and the injection direction is in horizontal, in which the analysis of the ray-tracing code and the better coupling of LH wave is considered. The transmission line is over-sized waveguide with low rf loss. (author)

  3. The pharmacological efficacy of the anti-IL17 scFv and sTNFR1 bispecific fusion protein in inflammation mouse stimulated by LPS.

    Science.gov (United States)

    Yang, Yongbi; Zhang, Teng; Cao, Hongxue; Yu, Dan; Zhang, Tong; Zhao, Shaojuan; Jing, Xiaohui; Song, Liying; Liu, Yunye; Che, Ruixiang; Liu, Xin; Li, Deshan; Ren, Guiping

    2017-08-01

    Acute lung injury (ALI) is still a leading cause of morbidity and mortality in critically ill patients. Recently, our study found that a bispecific fusion protein treatment can ameliorate the lung injury induced by LPS. However, the molecular mechanisms which bispecific fusion protein ameliorates acute lung injury remain unclear. In this study, we found that the bispecific fusion protein treatment inhibited the nuclear transcription of NF-κB in confocal laser scanning fluorescence microscopy, the bispecific fusion protein exert protective effects in the cell model of ALI induced by lipopolysaccharide (LPS) via inhibiting the nuclear factor κB (NF-κB) signaling pathway and mediate inflammation. Moreover, the treatment of the bispecific fusion protein show its efficacy in animal models stimulated by LPS, the results of real-time PCR and ELISA demonstrate that bispecific fusion protein treatment effectively inhibited the over-expression of inflammatory cytokines(tumor necrosis factor α, interleukin 1β and interleukin 17). In addition, LPS-challenged mice exhibited significant lung injury characterized by the deterioration of histopathology, which was meliorated by bispecific fusion protein treatment. Collectively, these results demonstrate that bispecific fusion protein treatment ameliorates LPS-induced ALI through reducing inflammatory cytokines and lung inflammation, which may be associated with the decreased the nuclear transcription of NF-κB. The bispecific fusion protein may be useful as a novel therapy to treat ALI. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

  4. Laser fusion systems design study. Final technical report

    International Nuclear Information System (INIS)

    1975-06-01

    This study investigated: (1) the formulation and evaluation of an alignment system to accomplish pointing, focusing, centering and translation for the 20-arm SHIVA laser, (2) the formulation and evaluation of concepts for the correction of static phase distortions introduced by the accumulated optical elements in the laser chains, (3) the formulation and evaluation of concepts for the correction of optical path length differences between the arms of the SHIVA system, and (4) the conceptual design of appropriate control system hardware. (U.S.)

  5. Respiratory syncytial virus subunit vaccine based on a recombinant fusion protein expressed transiently in mammalian cells.

    Science.gov (United States)

    Nallet, Sophie; Amacker, Mario; Westerfeld, Nicole; Baldi, Lucia; König, Iwo; Hacker, David L; Zaborosch, Christiane; Zurbriggen, Rinaldo; Wurm, Florian M

    2009-10-30

    Although respiratory syncytial virus (RSV) causes severe lower respiratory tract infection in infants and adults at risk, no RSV vaccine is currently available. In this report, efforts toward the generation of an RSV subunit vaccine using recombinant RSV fusion protein (rRSV-F) are described. The recombinant protein was produced by transient gene expression (TGE) in suspension-adapted human embryonic kidney cells (HEK-293E) in 4 L orbitally shaken bioreactors. It was then purified and formulated in immunostimulating reconstituted influenza virosomes (IRIVs). The candidate vaccine induced anti-RSV-F neutralizing antibodies in mice, and challenge studies in cotton rats are ongoing. If successful in preclinical and clinical trials, this will be the first recombinant subunit vaccine produced by large-scale TGE in mammalian cells.

  6. Oncogenic fusion proteins adopt the insulin-like growth factor signaling pathway.

    Science.gov (United States)

    Werner, Haim; Meisel-Sharon, Shilhav; Bruchim, Ilan

    2018-02-19

    The insulin-like growth factor-1 receptor (IGF1R) has been identified as a potent anti-apoptotic, pro-survival tyrosine kinase-containing receptor. Overexpression of the IGF1R gene constitutes a typical feature of most human cancers. Consistent with these biological roles, cells expressing high levels of IGF1R are expected not to die, a quintessential feature of cancer cells. Tumor specific chromosomal translocations that disrupt the architecture of transcription factors are a common theme in carcinogenesis. Increasing evidence gathered over the past fifteen years demonstrate that this type of genomic rearrangements is common not only among pediatric and hematological malignancies, as classically thought, but may also provide a molecular and cytogenetic foundation for an ever-increasing portion of adult epithelial tumors. In this review article we provide evidence that the mechanism of action of oncogenic fusion proteins associated with both pediatric and adult malignancies involves transactivation of the IGF1R gene, with ensuing increases in IGF1R levels and ligand-mediated receptor phosphorylation. Disrupted transcription factors adopt the IGF1R signaling pathway and elicit their oncogenic activities via activation of this critical regulatory network. Combined targeting of oncogenic fusion proteins along with the IGF1R may constitute a promising therapeutic approach.

  7. Application of Recommended Design Practices for Conceptual Nuclear Fusion Space Propulsion Systems

    Science.gov (United States)

    Williams, Craig H.

    2004-01-01

    An AIAA Special Project Report was recently produced by AIAA's Nuclear and Future Flight Propulsion Technical Committee and is currently in peer review. The Report provides recommended design practices for conceptual engineering studies of nuclear fusion space propulsion systems. Discussion and recommendations are made on key topics including design reference missions, degree of technological extrapolation and concomitant risk, thoroughness in calculating mass properties (nominal mass properties, weight-growth contingency and propellant margins, and specific impulse), and thoroughness in calculating power generation and usage (power-flow, power contingencies, specific power). The report represents a general consensus of the nuclear fusion space propulsion system conceptual design community and proposes 15 recommendations. This paper expands on the Report by providing specific examples illustrating how to apply each of the recommendations.

  8. Baculovirus display of fusion protein of Peste des petits ruminants virus and hemagglutination protein of Rinderpest virus and immunogenicity of the displayed proteins in mouse model

    International Nuclear Information System (INIS)

    Masmudur Rahman, Md.; Shaila, M.S.; Gopinathan, Karumathil P.

    2003-01-01

    Recombinant Bombyx mori nucleopolyhedroviruses (BmNPV) displaying the immunodominant ectodomains of fusion glycoprotein (F) of Peste des petitis ruminants virus (PPRV) and the hemagglutinin protein (H) of Rinderpest virus (RPV), on the budded virions as well as the surface of the infected host cells have been constructed. The F and H protein sequences were inserted in-frame within the amino-terminal region of BmNPV envelope glycoprotein GP64 expressing under the strong viral polyhedrin (polh) promoter. We improved the recombinant virus selection in BmNPV by incorporating the green fluorescent protein gene (gfp) as selection marker under a separate promoter within the transfer cassette harboring the desired genes. Following infection of the insect larvae or the host-derived BmN cells with these recombinant BmNPVs, the expressed GP64 fusion proteins were displayed on the host cell surface and the budded virions. The antigenic epitopes of the recombinant proteins were properly displayed and the recombinant virus particles induced immune response in mice against PPRV or RPV

  9. A conceptual design study of a reversed field pinch fusion reactor

    International Nuclear Information System (INIS)

    Kondo, S.; Tanaka, S.; Terai, T.; Hashizume, H.

    1989-01-01

    A conceptual design of a Reversed-Field Pinch (RFP) fusion reactor with a solid breeder blanket REPUTER-1 has been studied through parametric system studies and detailed design and analysis in order to clarify the technical feasibility of a compact fusion reactor. F-θ pumping is used for driving the plasma current necessary for steady state operation. A maintenance policy of replacing a whole fusion power core including TF coils is proposed to cope with the requirements of high wall loading and high mass power density. For the same reason a normal conductor is selected for most of the coils. The first wall is structurally independent of the blanket. The blanket module is composed of SiC reinforced blocks which form a stable arch so as to keep the stresses in SiC basically compressive. The coolant for the first wall and the limiter is pressurized water, while the coolant for the blanket is helium gas. A number of thin Li 2 O and thick beryllium tiles are packed into the blanket block so as to obtain a proper tritium breeding ratio. A pumped limiter is chosen for the plasma exhaust system. The study has shown the technical feasibility of a high power density fusion power reactor (330 kWe/tonne) with solid breeder blanket and many key physics and engineering issues are also clarified. (orig.)

  10. Conceptual capital-cost estimate and facility design of the Mirror-Fusion Technology Demonstration Facility

    International Nuclear Information System (INIS)

    1982-09-01

    This report contains contributions by Bechtel Group, Inc. to Lawrence Livermore National Laboratory (LLNL) for the final report on the conceptual design of the Mirror Fusion Technology Demonstration Facility (TDF). Included in this report are the following contributions: (1) conceptual capital cost estimate, (2) structural design, and (3) plot plan and plant arrangement drawings. The conceptual capital cost estimate is prepared in a format suitable for inclusion as a section in the TDF final report. The structural design and drawings are prepared as partial inputs to the TDF final report section on facilities design, which is being prepared by the FEDC

  11. Dynamic in vivo imaging and cell tracking using a histone fluorescent protein fusion in mice

    Directory of Open Access Journals (Sweden)

    Papaioannou Virginia E

    2004-12-01

    Full Text Available Abstract Background Advances in optical imaging modalities and the continued evolution of genetically-encoded fluorescent proteins are coming together to facilitate the study of cell behavior at high resolution in living organisms. As a result, imaging using autofluorescent protein reporters is gaining popularity in mouse transgenic and targeted mutagenesis applications. Results We have used embryonic stem cell-mediated transgenesis to label cells at sub-cellular resolution in vivo, and to evaluate fusion of a human histone protein to green fluorescent protein for ubiquitous fluorescent labeling of nucleosomes in mice. To this end we have generated embryonic stem cells and a corresponding strain of mice that is viable and fertile and exhibits widespread chromatin-localized reporter expression. High levels of transgene expression are maintained in a constitutive manner. Viability and fertility of homozygous transgenic animals demonstrates that this reporter is developmentally neutral and does not interfere with mitosis or meiosis. Conclusions Using various optical imaging modalities including wide-field, spinning disc confocal, and laser scanning confocal and multiphoton excitation microscopy, we can identify cells in various stages of the cell cycle. We can identify cells in interphase, cells undergoing mitosis or cell death. We demonstrate that this histone fusion reporter allows the direct visualization of active chromatin in situ. Since this reporter segments three-dimensional space, it permits the visualization of individual cells within a population, and so facilitates tracking cell position over time. It is therefore attractive for use in multidimensional studies of in vivo cell behavior and cell fate.

  12. Conceptual design of fusion experimental reactor (FER/ITER)

    International Nuclear Information System (INIS)

    Kimura, Haruyuki; Saigusa, Mikio; Saitoh, Yasushi

    1991-06-01

    Conceptual design of the Ion Cyclotron Wave (ICW) system for FER and Japanese contribution to the conceptual design of the ITER ICW system are presented. A frequency range of the FER ICW system is 50-85 MHz, which covers 2ω cT heating, current drive by transit time magnetic pumping (TTMP) and 2ω cD heating. Physics analyses show that the FER and the ITER ICW systems are suitable for the central ion heating and the burn control. The launching systems of the FER ICW system and the ITER high frequency ICW system are characterized by in-port plug and ridged-waveguide-fed 5x4 phased loop array. Merits of those systems are (1) a ceramic support is not necessary inside the cryostat and (2) remote maintenance of the front end part of the launcher is relatively easy. Overall structure of the launching system is consistent with radiation shielding, cooling, pumping, tritium safety and remote maintenance. The launcher has injection capability of 20 MW in the frequency range of 50-85 MHz with the separatrix-antenna distance of 15 cm and steep scrape-off density profile of H-mode. The shape of the ridged waveguide is optimized to provide desired frequency range and power handling capability with a finite element method. Matching between the current strap and the ridged waveguide is satisfactorily good. Thermal analysis of the Faraday shield shows that high electric conductivity low Z material such as beryllium should be chosen for a protection tile of the Faraday shield. Thick Faraday shield is necessary to tolerate electromagnetic force during disruptions. R and D needs for the ITER/FER ICW systems are identified and gain from JT-60/60U ICRF experiments and operations are indicated in connection with them. (author)

  13. Design of force-cooled conductors for large fusion magnets

    International Nuclear Information System (INIS)

    Dresner, L.; Lue, J.W.

    1977-01-01

    One type of conductor under consideration for tokamak toroidal field (TF) magnets is a cable-in-conduit cooled by supercritical helium in forced convection. The main problem is designing such force-cooled conductors (fcc) is to maintain adequate stability while keeping the pumping power tolerably low. The transit time of the helium through a coil is many minutes. Since recovery of the conductor from a thermomechanical perturbation takes on the order of tens of milliseconds, for purposes of calculation, the inventory of helium available to promote recovery is finite. This means that a large enough perturbation will quench the conductor. We can then judge the stability of a fcc by the maximum perturbation of some specified type against which the conductor is stable, i.e., can still return to the superconducting state. The simplest type of perturbation is a sudden, uniform heat input over the entire length of the conductor. The maximum, sudden, uniform heat input per unit volume of metal ΔH is called the ''stability margin.''

  14. A recombined fusion protein PTD-Grb2-SH2 inhibits the proliferation of breast cancer cells in vitro.

    Science.gov (United States)

    Yin, Jikai; Cai, Zhongliang; Zhang, Li; Zhang, Jian; He, Xianli; Du, Xilin; Wang, Qing; Lu, Jianguo

    2013-03-01

    The growth factor receptor bound protein 2 (Grb2) is one of the affirmative targets for cancer therapy, especially for breast cancer. In this study, we hypothesized the Src-homology 2 (SH2) domain in Grb2 may serve as a competitive protein-binding agent to interfere with the proliferation of breast cancer cells in vitro. We designed, constructed, expressed and purified a novel fusion protein containing the protein transduction domain (PTD) and Grb2-SH2 domain (we named it after PTD-Grb2-SH2). An immunofluorescence assay was used to investigate the location of PTD-Grb2-SH2 in cells. MTT assay and EdU experiments were applied to detect the proliferation of breast cancer cells. The ultra-structure was observed using transmission electron microscopy. Flow cytometry was used to determine the cytotoxicity of PTD-Grb2-SH2 on cell proliferation. We successfully obtained the PTD-Grb2-SH2 fusion protein in soluble form using a prokaryotic expression system. The new fusion protein successfully passed through both the cellular and nuclear membranes of breast cancer cells. The MTT assay showed that PTD-Grb2-SH2 exhibited significant toxicity to breast cancer cells in a dose- and time-dependent manner in vitro. EdU identified the decreased proliferation rates in treated MDA-MB-231 and SK-BR-3 cells. Observation by transmission electron microscopy and flow cytometry further confirmed the cytotoxicity as apoptosis. Our results show that the HIV1-TAT domain is a useful tool for transporting a low molecular weight protein across the cell membrane in vitro. The PTD-Grb2-SH2 may be a novel agent for breast cancer therapy.

  15. [Construction and prokaryotic expression of recombinant gene EGFRvIII HBcAg and immunogenicity analysis of the fusion protein].

    Science.gov (United States)

    Duan, Xiao-yi; Wang, Jian-sheng; Guo, You-min; Han, Jun-li; Wang, Quan-ying; Yang, Guang-xiao

    2007-01-01

    To construct recombinant prokaryotic expression plasmid pET28a(+)/c-PEP-3-c and evaluate the immunogenicity of the fusion protein. cDNA fragment encoding PEP-3 was obtained from pGEM-T Easy/PEP-3 and inserted into recombinant plasmid pGEMEX/HBcAg. Then it was subcloned in prokaryotic expression vector and transformed into E.coli BL21(DE3). The fusion protein was expressed by inducing IPTG and purified by Ni(2+)-NTA affinity chromatography. BALB/c mice were immunized with fusion protein and the antibody titre was determined by indirect ELISA. The recombinant gene was confirmed to be correct by restriction enzyme digestion and DNA sequencing. After prokaryotic expression, fusion protein existed in sediment and accounted for 56% of all bacterial lysate. The purified product accounted for 92% of all protein and its concentration was 8 g/L. The antibody titre in blood serum reached 1:16 000 after the fourth immunization and reached 1:2.56x10(5) after the sixth immunization. The titre of anti-PEP-3 antibody reached 1:1.28x10(5) and the titre of anti-HBcAg antibody was less than 1:4x10(3). Fusion gene PEP-3-HBcAg is highly expressed in E.coli BL21. The expressed fusion protein can induce neutralizing antibody with high titer and specificity, which lays a foundation for the study of genetically engineering vaccine for malignant tumors with the high expression of EGFRvIII.

  16. Conceptual design of a hybrid fusion-fission reactor with intrinsic safety and optimized energy productivity

    International Nuclear Information System (INIS)

    Talebi, Hosein; Sadat Kiai, S.M.

    2017-01-01

    Highlights: • Designing a high yield and feasible Dense Plasma Focus for driving the reactor. • Presenting a structural method to design the dual layer cylindrical blankets. • Finding, the blanket production energy, in terms of its geometrical and material parameters. • Designing a subcritical blanket with optimization of energy amplification in detail. - Abstract: A hybrid fission-fusion reactor with a Dense Plasma Focus (DPF) as a fusion core and the dual layer fissionable blanket as the energy multiplier were conceptually designed. A cylindrical DPF, energized by a 200 kJ bank energy, is considered to produce fusion neutron, and these neutrons drive the subcritical fission in the surrounding blankets. The emphasis has been placed on the safety and energy production with considering technical and economical limitations. Therefore, the k eff-t of the dual cylindrical blanket was defined and mathematically, specified. By applying the safety criterion (k eff-t ≤ 0.95), the geometrical and material parameters of the blanket optimizing the energy amplification were obtained. Finally, MCNPX code has been used to determine the detailed dimensions of the blankets and fuel rods.

  17. Safety methodology implementation in the conceptual design phase of a fusion reactor

    International Nuclear Information System (INIS)

    Rodriguez-Rodrigo, L.; Elbez-Uzan, J.

    2007-01-01

    The licensing of ITER in France represents the first process for licensing a fusion facility in the framework of an experimental device with a total Tritium inventory of 3 kg. The main ITER parameters are far from those expected in the future demonstration reactors where the fusion power will be at least 5 times higher and the additional heating power could also reach up to 5 times the one foreseen in ITER. Main safety requirements for these reactors are based, among other conditions, on their inherent features as low amount of fuel, very low impurity content of structural materials, minimum waste repository, no active systems for safe shut-down, and no need for evacuation of population after the most severe accident. The design of such reactors is at the stage of conceptual studies and is mainly dealing with plasma performances, tritium breeding, blanket/divertor designs and solution of engineering issues, as well as bounding accidents or classification of waste. The methodological approach for integrating safety analysis as a tool for optimizing the design of the overall fusion installation for future reactors in the conceptual design phase is sketched, including the machine itself and the different auxiliary nuclear buildings. (author)

  18. Numerical Design of Megawatt Gyrotron with 120 GHz Frequency and 50% Efficiency for Plasma Fusion Application

    Science.gov (United States)

    Kumar, Nitin; Singh, Udaybir; Kumar, Anil; Bhattacharya, Ranajoy; Singh, T. P.; Sinha, A. K.