Large non-circular RFP experiments at Wisconsin

International Nuclear Information System (INIS)

Sprott, J.C.; Dexter, R.N.; Prager, S.C.; Almagri, A.F.; Assadi, S.; Sarff, J.S.

1986-01-01

By removing the internal rings from the Levitated Octupole vacuum vessel, a large, non-circular RFP was produced. The major radius is 1.39 m, and the cross section is about 1 m/sup 2/. The device is unconventional in that the vacuum vessel, which consists of 5-cm thick aluminum with a single poloidal and toroidal gap, serves as the vacuum liner, conducting shell, and poloidal and toroidal field coils. A toroidal field of up to about 1 kG can be produced, and the poloidal field is driven by a 600 kJ capacitor bank through a 2-volt-second iron core. Discharges are initiated with 4200 volts per turn using self-reversal of the toroidal field in order to prevent arcing of the poloidal gap which is exposed to the plasma. The gap is protected with a 20-cm wide strip of ceramic. The best RFP discharges have a peak current of ≅200 kA and a duration of ≅ 10 msec. The toroidal field reverses when the current reaches ≅100 kA, making this one of the lowest current density RFP's in existence. The current ramps up to the final value over ≅10 resistive diffusion times and terminates only because the volt-second limit of the iron core is reached. The F-θ trajectory lies slightly to the right of the λ=constant theory as do all other RFP devices. Discharges have been produced with θ up to 2.5 and F as low as -0.8. A feature of the device is that it is capable of producing discharges with plasma current of ≅100 kA and ≅10 msec duration over a wide range of safety factor from the q>1 tokamak limit to the deeply-reversed, RFP limit. The highest current discharges (≅300 kA) are obtained at q≅0.5

Optimization of RFP formation and sustainment in RFX

Energy Technology Data Exchange (ETDEWEB)

Martini, S.; Buffa, A.; Collarin, P.; Lorenzi, A. De; Fiorentin, P.; Innocente, P.; Marchiori, G.; Paccagnella, R.; Piovan, R.; Sonato, P. [Istituto Gas Ionizzati del CNR, EURATOM-ENEA-CNR Association, Padua (Italy)

1993-12-31

In the first months of 1993, the RFX experiment (R=2 m, a=0.46 m) has operated at reduced volt-second (6 V{center_dot}s out of 15) to study the formation and sustainment of the RFP in a relatively safer power input regime, before increasing the parameters to reach the design value of 2 MA plasma current. At present the RFP configuration is obtained, similarly to the ETA-BETA II experiment, in the aided mode: a capacitor bank is discharged into the toroidal winding to produce an initial toroidal flux, {Phi}{sub T}, then the plasma current, I{sub T}, is induced by varying the poloidal flux stored in the magnetising winding; the free oscillation of the toroidal circuit continues and {Phi}{sub T} decays during the initial plasma current rise, until the toroidal field at the wall, B{sub T}(a), reverses and the toroidal circuit is crow-barred. The overall performance of the plasma during the RFP sustainment phase is strongly influenced by the control performed on density, toroidal field and plasma position during the formation phase. As soon as the RFP is obtained, a clear improvement of confinement is seen and the plasma current increases again until the applied toroidal voltage V{sub T}, which decreases exponentially, no longer matches the resistive drop. In RFX it is also possible to insert a pre-programmed flat-top power amplifier by which V{sub T} can be sustained and controlled in the range (0-60 V). In this way quasi-steady RFP current flat-top phases lasting {approx} 90 ms can be obtained which terminate only when the amplifiers are switched off and V{sub T} is no longer sustained. (author) 9 refs., 6 figs.

Optimization of RFP formation and sustainment in RFX

International Nuclear Information System (INIS)

Martini, S.; Buffa, A.; Collarin, P.; Lorenzi, A. De; Fiorentin, P.; Innocente, P.; Marchiori, G.; Paccagnella, R.; Piovan, R.; Sonato, P.

1993-01-01

In the first months of 1993, the RFX experiment (R=2 m, a=0.46 m) has operated at reduced volt-second (6 V·s out of 15) to study the formation and sustainment of the RFP in a relatively safer power input regime, before increasing the parameters to reach the design value of 2 MA plasma current. At present the RFP configuration is obtained, similarly to the ETA-BETA II experiment, in the aided mode: a capacitor bank is discharged into the toroidal winding to produce an initial toroidal flux, Φ T , then the plasma current, I T , is induced by varying the poloidal flux stored in the magnetising winding; the free oscillation of the toroidal circuit continues and Φ T decays during the initial plasma current rise, until the toroidal field at the wall, B T (a), reverses and the toroidal circuit is crow-barred. The overall performance of the plasma during the RFP sustainment phase is strongly influenced by the control performed on density, toroidal field and plasma position during the formation phase. As soon as the RFP is obtained, a clear improvement of confinement is seen and the plasma current increases again until the applied toroidal voltage V T , which decreases exponentially, no longer matches the resistive drop. In RFX it is also possible to insert a pre-programmed flat-top power amplifier by which V T can be sustained and controlled in the range (0-60 V). In this way quasi-steady RFP current flat-top phases lasting ∼ 90 ms can be obtained which terminate only when the amplifiers are switched off and V T is no longer sustained. (author) 9 refs., 6 figs

Engineering design of a compact RFP reactor (CRFPR)

International Nuclear Information System (INIS)

Hagenson, R.L.; Krakowski, R.A.

1983-01-01

The results of a previously-reported comprehensive parametric systems analysis of the Reversed-Field Pinch (RFP) give strong indications for minimum-cost systems that would operate with resistive water-cooled copper coils and higher first-wall neutron current (15 to 20 MW/m 2 ). These minimum-cost, compact RFP reactors (CRFPRs) have system power densities that can be comparable with fission power plants and, therefore, are 10 to 30 times smaller than most superconducting approaches. Reported herein are initial results of a conceptual engineering design of key fusion-power-core (FPC) subsystems

Over-relaxation phenomena during the set-up of RFP plasmas

International Nuclear Information System (INIS)

Nordlund, P.; Mazur, S.

1995-03-01

Experiments on the Extrap T1 reversed field pinch have shown that the formation of the RFP configuration is quite sensitive to the relative programming of the toroidal field and ohmic heating circuits. In this paper, new measurements of the evolution of the current density profile and of the spectral structure of the fluctuations during the set-up phase of RFP plasmas in the T1 experiment are presented. These measurements improve the understanding of the role of different spectral components in the dynamics of RFP formation. Under unfavourable (slow) set-up conditions, comparatively high energy is accumulated in m = 1 internal kinks prior to reversal of the edge toroidal fields. At reversal, nonlinearly driven m = 0 modes trigger a rapid broadening of the m = 1 spectrum. This behaviour is associated with a violent suppression of the current density in the core leading to an over-relaxation of the discharge involving a hollowing of the parallel current density profile. The over-relaxation phenomenon increases the volt-second consumption and plasma/wall interaction during RFP set-up, and degrades the flat-top discharge performance. 15 refs, 5 figs, 1 tab

Over-relaxation phenomena during the set-up of RFP plasmas

Energy Technology Data Exchange (ETDEWEB)

Nordlund, P.; Mazur, S.

1995-03-01

Experiments on the Extrap T1 reversed field pinch have shown that the formation of the RFP configuration is quite sensitive to the relative programming of the toroidal field and ohmic heating circuits. In this paper, new measurements of the evolution of the current density profile and of the spectral structure of the fluctuations during the set-up phase of RFP plasmas in the T1 experiment are presented. These measurements improve the understanding of the role of different spectral components in the dynamics of RFP formation. Under unfavourable (slow) set-up conditions, comparatively high energy is accumulated in m = 1 internal kinks prior to reversal of the edge toroidal fields. At reversal, nonlinearly driven m = 0 modes trigger a rapid broadening of the m = 1 spectrum. This behaviour is associated with a violent suppression of the current density in the core leading to an over-relaxation of the discharge involving a hollowing of the parallel current density profile. The over-relaxation phenomenon increases the volt-second consumption and plasma/wall interaction during RFP set-up, and degrades the flat-top discharge performance. 15 refs, 5 figs, 1 tab.

Expanding the Genetic Toolbox for Leptospira Species by Generation of Fluorescent Bacteria ▿

OpenAIRE

Aviat, Florence; Slamti, Leyla; Cerqueira, Gustavo M.; Lourdault, Kristel; Picardeau, Mathieu

2010-01-01

Our knowledge of the genetics and molecular basis of the pathogenesis associated with Leptospira, in comparison to those of other bacterial species, is very limited. An improved understanding of pathogenic mechanisms requires reliable genetic tools for functional genetic analysis. Here, we report the expression of gfp and mRFP1 genes under the control of constitutive spirochetal promoters in both saprophytic and pathogenic Leptospira strains. We were able to reliably measure the fluorescence ...

The reduction of low frequency fluctuations in RFP experiments

International Nuclear Information System (INIS)

Phillips, J.A.; Baker, D.A.; Gribble, R.F.

1998-01-01

The low frequency fluctuations seen in RFP experiments are found to be correlated with changes in the toroidal flux measured by diamagnetic loops surrounding the discharge. The correlation of the onset of impurity radiation and x-rays with the crash seen in experiments is caused by plasma bombarding the metal liner associated with this loss of flux. Efforts should be made to design improved stabilizing shells that will reduce the loss of flux and give improved RFP energy confinement times

The first results of siliconization on SWIP-RFP device

International Nuclear Information System (INIS)

Zhang Peng; Li Qiang; Luo Cuiwen; Li Jieping; Qian Shangjie; Fang Shuiquan; Yi Ping; Xue Jun; Li Kehua; Luo Junlin; Hong Wenyu; Cao Zeng; Zhang Nianman; Wang Quanming; Li Jie; Huang Ming; Zhong Yunze; Zhang Qingchun; Luo Cuixian

1997-01-01

The first results of reversed field pinch (RFP) and ultra low safety factor (ULQ) plasma experiments with siliconization on SWIP-RFP device are presented in this paper. The siliconization decreases the impurity concentrations in the plasma and increases the configuration sustainment time. Ion temperature has been estimated with the CV line of the visible light spectra and the broadening of CIII lines in vacuum ultraviolet (VUV) region. The anomalous ion heating as well as the anomalous resistance were observed. (orig.)

Fusion core start-up, ignition, and burn simulations of reversed-field pinch (RFP) reactors

International Nuclear Information System (INIS)

Chu, Y.Y.

1988-01-01

A transient reactor simulation model is developed to investigate and simulate the start-up, ignition, and burn of a reversed-field pinch reactor. The simulation is based upon a spatially averaged plasma balance model with field profiles obtained from MHD quasi-equilibrium analysis. Alpha particle heating is estimated from Fokker-Planck calculations. The instantaneous plasma current is derived from a self-consistent circuit analysis for plasma/coil/eddy current interactions. The simulation code is applied to the TITAN RFP reactor design which features a compact, high-power-density reversed-field pinch fusion system. A contour analysis is performed using the steady-state global plasma balance. The results are presented with contours of constant plasma current. A saddle point is identified in the contour plot which determined the minimum value of plasma current required to achieve ignition. In the simulations of the TITAN RFP reactor, the OH-driven super-conducting EF coils are found to deviate from the required equilibrium values as the induced plasma current increases. A set of basic results from the simulation of TITAN RFP reactor yield a picture of RFP plasma operation in a reactor. Investigations of eddy currents are also presented and have very important in reactor design

Novel tumorigenic rearrangement, Δrfp/ret, in a papillary thyroid carcinoma from externally irradiated patient

International Nuclear Information System (INIS)

Saenko, Vladimir; Rogounovitch, Tatiana; Shimizu-Yoshida, Yuki; Abrosimov, Aleksandr; Lushnikov, Eugeny; Roumiantsev, Pavel; Matsumoto, Naomichi; Nakashima, Masahiro; Meirmanov, Serik; Ohtsuru, Akira; Namba, Hiroyuki; Tsyb, Anatoly; Yamashita, Shunichi

2003-01-01

Molecular analysis of cDNA derived from a papillary thyroid carcinoma (PTC) (follicular variant of papillary thyroid carcinoma on histology) which developed in an externally irradiated patient 4 years after exposure identified a portion of the 5' region, exons 1-3, of the rfp gene juxtaposed upstream of the fragment encoding the tyrosine kinase (TK) domain of the ret gene. The fusion gene, termed Δrfp/ret, was the result of a balanced chromosomal translocation t(6;10) (p21.3;q11.2) confirmed by interphase FISH painting, with breakpoints occurring in introns 3 and 11 of the rfp and ret genes, respectively. Both Δrfp/ret and reciprocal ret/rfp chimeric introns had small deletions around breakpoints consistent with presumed misrepair of a radiation-induced double-strand DNA break underlying the rearrangement. No extensive sequence homology was found between the fragments flanking the breakpoints. The fusion protein retained the propensity to form oligomers likely to be mediated by a coiled-coil of the RFP polypeptide as assessed by a yeast two-hybrid system. NIH 3T3 fibroblasts stably transfected with a mammalian expression vector encoding full-length ΔRFP/RET readily gave rise to the tumors in athymic mice suggestive of high transforming potential of the fusion protein. Thus, the Δrfp/ret rearrangement may be causatively involved in cancerogenesis and provides additional evidence of the role of activated ret oncogene in the development of a subset of papillary thyroid carcinoma

Novel tumorigenic rearrangement, {delta}rfp/ret, in a papillary thyroid carcinoma from externally irradiated patient

Energy Technology Data Exchange (ETDEWEB)

Saenko, Vladimir; Rogounovitch, Tatiana; Shimizu-Yoshida, Yuki; Abrosimov, Aleksandr; Lushnikov, Eugeny; Roumiantsev, Pavel; Matsumoto, Naomichi; Nakashima, Masahiro; Meirmanov, Serik; Ohtsuru, Akira; Namba, Hiroyuki; Tsyb, Anatoly; Yamashita, Shunichi

2003-06-19

Molecular analysis of cDNA derived from a papillary thyroid carcinoma (PTC) (follicular variant of papillary thyroid carcinoma on histology) which developed in an externally irradiated patient 4 years after exposure identified a portion of the 5' region, exons 1-3, of the rfp gene juxtaposed upstream of the fragment encoding the tyrosine kinase (TK) domain of the ret gene. The fusion gene, termed {delta}rfp/ret, was the result of a balanced chromosomal translocation t(6;10) (p21.3;q11.2) confirmed by interphase FISH painting, with breakpoints occurring in introns 3 and 11 of the rfp and ret genes, respectively. Both {delta}rfp/ret and reciprocal ret/rfp chimeric introns had small deletions around breakpoints consistent with presumed misrepair of a radiation-induced double-strand DNA break underlying the rearrangement. No extensive sequence homology was found between the fragments flanking the breakpoints. The fusion protein retained the propensity to form oligomers likely to be mediated by a coiled-coil of the RFP polypeptide as assessed by a yeast two-hybrid system. NIH 3T3 fibroblasts stably transfected with a mammalian expression vector encoding full-length {delta}RFP/RET readily gave rise to the tumors in athymic mice suggestive of high transforming potential of the fusion protein. Thus, the {delta}rfp/ret rearrangement may be causatively involved in cancerogenesis and provides additional evidence of the role of activated ret oncogene in the development of a subset of papillary thyroid carcinoma.

RFP for the Auroral Multiscale Midex (AMM) Mission star tracker

DEFF Research Database (Denmark)

Riis, Troels; Betto, Maurizio; Jørgensen, John Leif

1999-01-01

This document is in response to the John Hopkins University - Applied Physics Laboratory RFP for the Auroral Multiscale Midex Mission star tracker.It describes the functionality, the requirements and the performance of the ASC Star Tracker.......This document is in response to the John Hopkins University - Applied Physics Laboratory RFP for the Auroral Multiscale Midex Mission star tracker.It describes the functionality, the requirements and the performance of the ASC Star Tracker....

Gateway binary vectors with the bialaphos resistance gene, bar, as a selection marker for plant transformation.

Science.gov (United States)

Nakamura, Shinya; Mano, Shoji; Tanaka, Yuji; Ohnishi, Masato; Nakamori, Chihiro; Araki, Masami; Niwa, Tomoko; Nishimura, Mikio; Kaminaka, Hironori; Nakagawa, Tsuyoshi; Sato, Yutaka; Ishiguro, Sumie

2010-01-01

We constructed two series of Gateway binary vectors, pGWBs and R4pGWBs, possessing the bialaphos resistance gene (bar) as a selection marker for plant transformation. The reporters and tags employed in this system are sGFP, GUS, LUC, EYFP, ECFP, G3GFP, mRFP, TagRFP, 6xHis, FLAG, 3xHA, 4xMyc, 10xMyc, GST, T7 and TAP. Selection of Arabidopsis transformants with BASTA was successfully carried out using both plate-grown and soil-grown seedlings. Transformed rice calli and suspension-cultured tobacco cells were selected on plates containing BASTA or glufosinate-ammonium. These vectors are compatible with existing pGWB and R4pGWB vectors carrying kanamycin and hygromycin B resistance.

The RFP, A New Management Device for Pennsylvania

Science.gov (United States)

Jeffers, Donna A.

1976-01-01

Available from: EC 090 474. Reviewed are the guidelines set forth in the RFP (Request for Proposal) document regarding educational programs for adults and juveniles incarcerated in Pennsylvania institutions. (SBH)

Initial results from the rebuilt EXTRAP T2R RFP device

Science.gov (United States)

Brunsell, P. R.; Bergsåker, H.; Cecconello, M.; Drake, J. R.; Gravestijn, R. M.; Hedqvist\\ad{2 }, A.; Malmberg, J.-A.

2001-11-01

The EXTRAP T2R thin shell reversed-field pinch (RFP) device has recently resumed operation after a major rebuild including the replacement of the graphite armour with molybdenum limiters, a fourfold increase of the shell time constant, and the replacement of the helical coil used for the toroidal field with a conventional solenoid-type coil. Wall-conditioning using hydrogen glow discharge cleaning was instrumental for successful RFP operation. Carbon was permanently removed from the walls during the first week of operation. The initial results from RFP operation with relatively low plasma currents in the range Ip = 70-100 kA are reported. RFP discharges are sustained for more than three shell times. Significant improvements in plasma parameters are observed, compared to operation before the rebuild. There is a substantial reduction in the carbon impurity level. The electron density behaviour is more shot-to-shot reproducible. The typical density is ne = 0.5-1×1019 m-3. Monitors of Hα line radiation indicate that the plasma wall interaction is more toroidally symmetric and that there is less transient gas release from the wall. The minimum loop voltage is in the range Vt = 28-35 V, corresponding to a reduction by a factor of two to three compared to the value before the rebuild.

New chicken Rfp-Y haplotypes on the basis of MHC class II RFLP and MLC analyses

DEFF Research Database (Denmark)

Juul-Madsen, H R; Zoorob, R; Auffray, C

1997-01-01

New chicken Rfp-Y haplotypes were determined by the use of restriction fragment length polymorphism (RFLP) and mixed lymphocyte culture (MLC) in four different chicken haplotypes, B15, B19, B21, B201. The RFLP polymorphism was mapped to the Rfp-Y system by the use of a subclone (18.1) which maps...... near a polymorphic lectin gene located in the Rfp-Y system and DNA from families with known segregation of the implicated RFLP polymorphism. For the first time it is shown that major histocompatibility complex class II genes in the Rfp-Y system have functional implications. Sequence information...

75 FR 40856 - Federal Register Meeting Notice; Webinar About Regional Innovation Clusters RFP

Science.gov (United States)

2010-07-14

... potential Offerors about the Regional Innovation Clusters RFP. For more information please go to http://www.sba.gov/clusters/index.html . The RFP may be found on http://www.fedbizopps.gov . Logistical Information: The webinar will be held on Thursday, July 15, 2010. For details, please visit http://www.sba.gov...

Scrape-off model and pumped-limiter design for reversed-field pinches (RFP)

International Nuclear Information System (INIS)

Embrechts, M.J.; Bathke, C.G.; Krakowski, R.A.

1983-01-01

In order to develop a better understanding of the plasma/first-wall interaction in an RFP configuration, the models being developed to describe edgeplasma and scrapeoff regions for the tokamak have been adopted. Specifically, a scrapeoff model similar to the one developed and used for the tokamak FED design is used to model the parameter range where pumped limiters may be applied to the compact RFP. The one-dimensional, steady-state heat and particle equations are solved in the scrapeoff layer for an RFP geometry, assuming equal electron and ion temperature and density, convective transport along field lines, and Bohm-like diffusion. All charge-exchange and radiation effects are assumed to take place in the region between plasma and scrapeoff layer, and only a specific fraction of the total plasma energy will enter the scrapeoff layer in the form of conduction and convection. A simplified recycling model based on an effective recycling coefficient is used. For a given particle and energy flux entering the scrapeoff and for specified relationships between the particle flux and the energy flux near the first wall, the temperature and density profiles in the scrapeoff layer region are determined. The shape of the limiter surface is determined iteratively for a specified number of poloidal limiters by specifying a constant (design) heat flux for the limiter surfaces

Effect of secretory pathway gene overexpression on secretion of a fluorescent reporter protein in Aspergillus nidulans

DEFF Research Database (Denmark)

Schalén, Martin; Anyaogu, Diana Chinyere; Hoof, Jakob Blæsbjerg

2016-01-01

roles in the process have been identified through transcriptomics. The assignment of function to these genes has been enabled in combination with gene deletion studies. In this work, 14 genes known to play a role in protein secretion in filamentous fungi were overexpressed in Aspergillus nidulans....... The background strain was a fluorescent reporter secreting mRFP. The overall effect of the overexpressions could thus be easily monitored through fluorescence measurements, while the effects on physiology were determined in batch cultivations and surface growth studies. Results: Fourteen protein secretion...... pathway related genes were overexpressed with a tet-ON promoter in the RFP-secreting reporter strain and macromorphology, physiology and protein secretion were monitored when the secretory genes were induced. Overexpression of several of the chosen genes was shown to cause anomalies on growth, micro...

Refueling and control of RFP burns

International Nuclear Information System (INIS)

Nebel, R.; Miley, G.H.

1978-01-01

An earlier study of the stability of a fusion burn in a Reversed Field Pinch (RFP) has been extended to include cold particle refueling. This refueling, coupled with anomalous transport, makes possible quasi-steady state operation which both flattens the wall-loading temporal dependence and significantly increases energy gain factors. This paper discusses results of these burn simulations along with parametric studies aimed at determining associated reactor scaling problems

Next-Step scientific objectives, targets, and parameters for reversed-field-pinch (RFP) magnetic fusion energy (MFE) systems: Preliminary thoughts

International Nuclear Information System (INIS)

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

1993-01-01

The purpose of this document is the quantitative definition of objectives, targets, and parameters of the Next-Step device to follow the present RFX experiment; this device is given the name RFXNS. Although developed over five years ago, much of the material distilled into the 1988 RFP tactical plan is useful in establishing the goals and parameters of RFXNS. This earlier plan established tentative parameters of an RFP next step based on: predictions of RFP ignition and commercial-reactor devices; and the assumed successful operation of highly complementary RFP experiments RFX and ZTH/CPRF. Programmatic changes and evolution that have occurred since 1988 strongly impact the role and characteristics of an RFXNS: the Los Alamos ZTH/CPRF project and fusion program was terminated in mid-construction for reasons of MFE cost savings and concept focusing; great progress has been made in launching ITER; and reactor projections for the tokamak have increased in detail and variety, but not in commercial promise and competitiveness. A brief status of and perspective from each of the above three points is necessary before the key issues and their implementation to form the basis of the RFXNS definition are given

Efektivitas penggunaan gambir sebagai bahan penyamak nabati sistem C-RFP untuk pembuatan kulit jaket dari kulit domba

OpenAIRE

Sri Sutyasmi

2017-01-01

Gambier is obtained from the extraction of leaves and young twigs of Uncaria gambir, a shrub that grows in Indonesia. Gambier contains tannin by 20% to 55% so that can be used as a vegetable tanning material using the C-RFP system (C=Conditioning, R=Rapid, F=Fass (drum), P=Powder). This research aimed to determine the effectiveness of gambier as a vegetable tanning material using the C-RFP system. Pickled leather underwent a tanning process using the C-RFP system with gambier as the tanning m...

Efektivitas Penggunaan Gambir Sebagai Bahan Penyamak Nabati Sistem C-RFP Untuk Pembuatan Kulit Jaket Dari Kulit Domba

OpenAIRE

Sutyasmi, Sri

2017-01-01

Gambier is obtained from the extraction of leaves and young twigs of Uncaria gambir, a shrub that grows in Indonesia. Gambier contains tannin by 20% to 55% so that can be used as a vegetable tanning material using the C-RFP system (C=Conditioning, R=Rapid, F=Fass (drum), P=Powder). This research aimed to determine the effectiveness of gambier as a vegetable tanning material using the C-RFP system. Pickled leather underwent a tanning process using the C-RFP system with gambier as the tanning m...

Restriction Fragment Pattern (RFP) analysis of genomes from Danish isolates of Suid herpesvirus 1 (Aujeszky's disease virus)

DEFF Research Database (Denmark)

Christensen, Laurids Siig; Sørensen, K. J.; Lei, J. C.

1987-01-01

Purified DNA from 42 isolates of Suid herpesvirus 1 (SHV-1) collected during 1985 from clinical outbreaks of Aujezsky's disease on Danish farms was compared by restriction fragment pattern (RFP) analysis. The BamHI generated RFPs were found to be distinguishable, thus confirming RFP analysis...

A monomeric variant of insulin degrading enzyme (IDE loses its regulatory properties.

Directory of Open Access Journals (Sweden)

Eun Suk Song

2010-03-01

Full Text Available Insulin degrading enzyme (IDE is a key enzyme in the metabolism of both insulin and amyloid beta peptides. IDE is unique in that it is subject to allosteric activation which is hypothesized to occur through an oligomeric structure.IDE is known to exist as an equilibrium mixture of monomers, dimers, and higher oligomers, with the dimer being the predominant form. Based on the crystal structure of IDE we deleted the putative dimer interface in the C-terminal region, which resulted in a monomeric variant. Monomeric IDE retained enzymatic activity, however instead of the allosteric behavior seen with wild type enzyme it displayed Michaelis-Menten kinetic behavior. With the substrate Abz-GGFLRKHGQ-EDDnp, monomeric IDE retained approximately 25% of the wild type activity. In contrast with the larger peptide substrates beta-endorphin and amyloid beta peptide 1-40, monomeric IDE retained only 1 to 0.25% of wild type activity. Unlike wild type IDE neither bradykinin nor dynorphin B-9 activated the monomeric variant of the enzyme. Similarly, monomeric IDE was not activated by polyphosphates under conditions in which the activity of wild type enzyme was increased more than 50 fold.These findings serve to establish the dimer interface in IDE and demonstrate the requirement for an oligomeric form of the enzyme for its regulatory properties. The data support a mechanism where the binding of activators to oligomeric IDE induces a conformational change that cannot occur in the monomeric variant. Since a conformational change from a closed to a more open structure is likely the rate-determining step in the IDE reaction, the subunit induced conformational change likely shifts the structure of the oligomeric enzyme to a more open conformation.

Monomeric Yeast Frataxin is an Iron-Binding Protein

International Nuclear Information System (INIS)

Cook, J.; Bencze, K.; Jankovic, A.; Crater, A.; Busch, C.; Bradley, P.; Stemmler, A.; Spaller, M.; Stemmler, T.

2006-01-01

Friedreich's ataxia, an autosomal cardio- and neurodegenerative disorder that affects 1 in 50 000 humans, is caused by decreased levels of the protein frataxin. Although frataxin is nuclear-encoded, it is targeted to the mitochondrial matrix and necessary for proper regulation of cellular iron homeostasis. Frataxin is required for the cellular production of both heme and iron-sulfur (Fe-S) clusters. Monomeric frataxin binds with high affinity to ferrochelatase, the enzyme involved in iron insertion into porphyrin during heme production. Monomeric frataxin also binds to Isu, the scaffold protein required for assembly of Fe-S cluster intermediates. These processes (heme and Fe-S cluster assembly) share requirements for iron, suggesting that monomeric frataxin might function as the common iron donor. To provide a molecular basis to better understand frataxin's function, we have characterized the binding properties and metal-site structure of ferrous iron bound to monomeric yeast frataxin. Yeast frataxin is stable as an iron-loaded monomer, and the protein can bind two ferrous iron atoms with micromolar binding affinity. Frataxin amino acids affected by the presence of iron are localized within conserved acidic patches located on the surfaces of both helix-1 and strand-1. Under anaerobic conditions, bound metal is stable in the high-spin ferrous state. The metal-ligand coordination geometry of both metal-binding sites is consistent with a six-coordinate iron-(oxygen/nitrogen) based ligand geometry, surely constructed in part from carboxylate and possibly imidazole side chains coming from residues within these conserved acidic patches on the protein. On the basis of our results, we have developed a model for how we believe yeast frataxin interacts with iron

Simulation study of MHD relaxation and reconnection processes in RFP plasma

International Nuclear Information System (INIS)

Kusano, Kanya; Kunimoto, Kaito; Suzuki, Yoshio; Tamano, Teruo; Sato, Tetsuya

1991-01-01

The authors have studied several nonlinear processes in RFP plasma through the use of 3D MHD simulations. In particular, they have shed light on: (1) dynamo and self-sustainment in reversed-field pinch (RFP), (2) phase locking process in MHD relaxation, and (3) the heating and acceleration in magnetic reconnection process. First, the contributions of the kink (m = 1) mode (linearly unstable) and of the m = 0 mode (driven by nonlinear coupling) to the dynamo are qualitatively evaluated using a high accuracy simulation. It is found that, if the free energy to drive kink instabilities is as small as that in the actual experimental plasma, the m = 0 modes, driven nonlinearly, play a more important role for the flux generation than the kink modes. Secondly, numerical simulations of the self-sustainment process in a RFP are performed. It is confirmed that the self-sustainment process is a coherent oscillating process composed of the MHD relaxation and the resistive diffusion processes. Toroidal phase locking process of kink modes is numerically observed in simulations of self-reversal and self-sustainment processes. It has characteristics similar to the slinky mode observed in the OHTE experiment. A detailed investigation reveals that nonlinear coupling between the most unstable two kink modes governs the entire dynamics in all kink modes and leads to the phase locking process. They find that reconnection can accelerate plasma over a local Alfven speed. This is a result of the fact that the magnetic field in the downstream area plays a similar role to de Laval nozzle. They also investigate the heating mechanisms in reconnection process. It is revealed that the viscous heating rate is as large as the joule heating rate in the reconnection process. This result implies that the viscous heating in the reconnection process is an important candidate for the mechanism to explain the RFP experiments where the ion temperatures is higher than the electron temperature

Magnetic measurements for RFP experiment on STP-3(M)

International Nuclear Information System (INIS)

Nagata, Akiyoshi; Tamaru, Takeshi; Arimoto, Hideki; Yamada, Shuichi; Sato, Koichi.

1984-03-01

Magnetic measurements are arranged for RFP experiment on STP-3(M). Magnetic measurements will be applied to investigate the discharge parameters, F(field reversal ratio) - theta(pinch parameter) diagram, the physical mechanism of flux enhancement and the toroidal and poloidal mode numbers due to the MHD instability. Theoretical considerations and instrumental techniques for magnetic measurements are described in detail. (author)

Electron transport in the stochastic fields of RFP ZT-40M

International Nuclear Information System (INIS)

Punjabi, A.; Verma, A.; Kim, Myung-Hee; Boozer, A.

1991-01-01

In this paper, we use our newly developed Monte Carlo Method to study the transport of electrons in stochastic magnetic fields of the device RFP ZT4OM. The results of this calculation will be compared with the Rechester-Rosenbluth scaling

[Establishment of an iRFP and luciferase dual-color fluorescence-traced hepatocellular carcinoma transplantation model in nude mice].

Science.gov (United States)

Li, Hongjun; Yang, Tianhua; Huang, Yanping; Liu, Mingzhu; Qin, Zhongqiang; Chu, Fei; Li, Zhenghong; Li, Yonghai

2017-11-01

Objective To establish a hepatocellular carcinoma xenograft model in nude mice which could stably express gene and be monitored dynamically. Methods We first constructed the lentiviral particles containing luciferase (Luc) and near-infrared fluorescent protein (iRFP) and puromycin resistance gene, and then transduced them into the HepG2 hepatoma cells. The cell line stably expressing Luc and iRFP genes were screened and inoculated into nude mice to establish xenograft tumor model. Tumor growth was monitored using in vivo imaging system. HE staining and immunohistochemistry were used to evaluate the pathological features and tumorigenic ability. Results HepG2 cells stably expressing iRFP and Luc were obtained; with the engineered cell line, xenograft model was successfully established with the features of proper tumor developing time and high rate of tumor formation as well as typical pathological features as showed by HE staining and immunohistochemistry. Conclusion Hepatocellular carcinoma model in nude mice with the features of stable gene expression and dynamical monitoring has been established successfully with the HepG2-iRFP-Luc cell line.

Confinement properties of the RFP [Reversed Field Pinch

International Nuclear Information System (INIS)

Weber, P.G.; Schoenberg, K.F.; Ingraham, J.C.; Miller, G.; Munson, C.P.; Pickrell, M.M.; Wurden; Tsui, H.Y.W.; Ritz, Ch.P.

1990-01-01

Research in ZT-40M has been focused on elucidating the confinement properties of the Reversed Field Pinch (RFP). Recent improvements in diagnostic capability have permitted measurement of radial profiles, as well as a detailed study of the edge plasma. The emerging confinement picture for ZT-40M has several ingredients: Typically 0.3 of the Ohmic input power to ZT-40M is available to drive fluctuations. Evidence points to this fluctuational power heating the ions. Approximately one quarter of the input power is lost through radiation, with metal impurities playing a key role. Magnetic fluctations in ZT-40M are at the percent level, as measured in the edge plasma. Extrapolating these data to small radii shows stochasticity in the core plasma. Suprathermal electrons are measured in the edge plasma. These electrons originate in the core, and transport to the edge along the fluctuating magnetic field lines. Under typical conditions, these electrons constitute the major electron energy loss channel in ZT-40M. Electrostatic fluctuations dominate the edge electron particle flux, but not the electron thermal flux. The major ion loss process is charge exchange, with smaller contributions from conduction and convection. In examining these observations, and the parametric dependences of confinement, a working model for RFP confinement emerges. An overview of this model, together with implications for the multi-mega-ampere ZTH experiment will be presented

The PDZ and band 4.1 containing protein Frmpd1 regulates the subcellular location of activator of G-protein signaling 3 and its interaction with G-proteins.

Science.gov (United States)

An, Ningfei; Blumer, Joe B; Bernard, Michael L; Lanier, Stephen M

2008-09-05

Activator of G-protein signaling 3 (AGS3) is one of nine mammalian proteins containing one or more G-protein regulatory (GPR) motifs that stabilize the GDP-bound conformation of Galphai. Such proteins have revealed unexpected functional diversity for the "G-switch" in the control of events within the cell independent of the role of heterotrimeric G-proteins as transducers for G-protein-coupled receptors at the cell surface. A key question regarding this class of proteins is what controls their subcellular positioning and interaction with G-proteins. We conducted a series of yeast two-hybrid screens to identify proteins interacting with the tetratricopeptide repeat (TPR) of AGS3, which plays an important role in subcellular positioning of the protein. We report the identification of Frmpd1 (FERM and PDZ domain containing 1) as a regulatory binding partner of AGS3. Frmpd1 binds to the TPR domain of AGS3 and coimmunoprecipitates with AGS3 from cell lysates. Cell fractionation indicated that Frmpd1 stabilizes AGS3 in a membrane fraction. Upon cotransfection of COS7 cells with Frmpd1-GFP and AGS3-mRFP, AGS3-mRFP is observed in regions of the cell cortex and also in membrane extensions or processes where it appears to be colocalized with Frmpd1-GFP based upon the merged fluorescent signals. Frmpd1 knockdown (siRNA) in Cath.a-differentiated neuronal cells decreased the level of endogenous AGS3 in membrane fractions by approximately 50% and enhanced the alpha2-adrenergic receptor-mediated inhibition of forskolin-induced increases in cAMP. The coimmunoprecipitation of Frmpd1 with AGS3 is lost as the amount of Galphai3 in the cell is increased and AGS3 apparently switches its binding partner from Frmpd1 to Galphai3 indicating that the interaction of AGS3 with Frmpd1 and Galphai3 is mutually exclusive. Mechanistically, Frmpd1 may position AGS3 in a membrane environment where it then interacts with Galphai in a regulated manner.

MHD activity during RFP startup in Extrap T1

International Nuclear Information System (INIS)

Drake, J.R.; Brzozowski, J.; Mazur, S.; Nordlund, P.

1993-04-01

During the startup phase of a reversed field pinch discharge (RFP), the configuration undergoes relaxation which is the dynamic process by which the configuration evolves to states near the minimum magnetic energy thus realising improved stability. A high loop voltage and a high level of level of m =1 poloidal mode activity is observed while this process occurs. After the RFP configuration is established, there is a substantial decrease in both the m = 1 activity and the loop voltage. In addition in the Extrap T1 device, under unfavourable conditions characterised by a pinch parameter θ less than about 1.5 at the time of reversal, the loop voltage can further increase substantially resulting in a suppressed plasma current level. Simultaneous with the loop voltage increase there is an observed increase in poloidal m = 0 mode activity. This occurs just when field reversal first is established as the surface with zero toroidal field is produced at the plasma edge. The increased loop voltage is believed to be the result of m = 0 tearing modes resonant at the reversal surface and driven by a large parallel current gradient at the edge which become particularly pathological when the θ, is comparatively low

Pengaruh mimosa pada penyamakan kulit jaket domba samak nabati menggunakan sistem C-RFP, ditinjau dari sifat organoleptis, fisis, dan morfologi kulit

Directory of Open Access Journals (Sweden)

Sri Sutyasmi

2016-06-01

Full Text Available The purpose of this study was to determine the effect of the use of mimosa, to manufacture environmentally friendly leather for jacket by using the C-RFP system (C=Conditioning, R=Rapid, F=Fass (drum, P=Powder, on physical, organoleptic, and morphology properties of leather. It was also to obtain a formula for vegetable tanning with C-RFP system. Pickled skins are conditioned (pre-tanning using Sodotan TSN and Sodotan APR, and then Sodotan TSN was chosen due to it meets the requirements of SNI leather for jacket (SNI 4593:2011. Then, pickled skins were tanned with mimosa and applied C-RFP system or rapid tanning without water added. Mimosa, used in this research, were 15%, 20%, and 25% and fatliquor were 12.5%, 15%, and 17.5%. Furthermore, The leather were finished into an environmentally friendly leather for jackets, and then tested for physical and organoleptic properties based on the SNI 4593:2011 as well as leather morphology (SEM. The physical test result shows that for mimosa 15%, 20% and 25%, and for fatliquor 15% and 17.5% are fullfill the SNI.

Efektivitas penggunaan gambir sebagai bahan penyamak nabati sistem C-RFP untuk pembuatan kulit jaket dari kulit domba

Directory of Open Access Journals (Sweden)

Sri Sutyasmi

2017-06-01

Full Text Available Gambier is obtained from the extraction of leaves and young twigs of Uncaria gambir, a shrub that grows in Indonesia. Gambier contains tannin by 20% to 55% so that can be used as a vegetable tanning material using the C-RFP system (C=Conditioning, R=Rapid, F=Fass (drum, P=Powder. This research aimed to determine the effectiveness of gambier as a vegetable tanning material using the C-RFP system. Pickled leather underwent a tanning process using the C-RFP system with gambier as the tanning material at various percentages, namely by 15%, 20%, and 25%, to be processed as leather for jackets. The variations of oil used to keep the leather supple were 12.5%, 15%, and 17.5%. The tanned leather was then tested in terms of its organoleptic, physical, chemical, and morphological properties. Results of the organoleptic test and most of the results of the physical test showed that leather for jackets which was tanned using a vegetable tanning material met the requirements set out by SNI 4593: 2011 (sheepskin/goatskin leather for jackets. Results of the SEM test indicated that leather for jackets which was tanned using a vegetable tanning material displayed dense and strong skin tissue, and this proves that gambier is very effective for vegetable tanning using the C-RFP system.

Measurements of fast ion spatial dynamics during magnetic activity in the RFP

Science.gov (United States)

Goetz, J. A.; Anderson, J. K.; Bonofiglo, P.; Kim, J.; McConnell, R.; Magee, R. M.

2017-10-01

Fast ions in the RFP are only weakly affected by a stochastic magnetic field and behave nearly classically in concentration too low to excite Alfvenic activity. At high fast ion concentration sourced by H-NBI in 300kA RFP discharges, a substantial drop in core-localized high pitch fast ions is observed during bursts of coupled EPM and IAE (magnetic island-induced Alfven eigenmode) activity (100-200kHz) through neutral particle analysis. Sourcing instead fast deuterium with NBI, the DD fusion products can measure the dynamics of the fast ion density profile. Both a collimated neutron detector and a new 3MeV fusion proton detector loaned by TriAlpha Energy measure the fast ion density profile with 5cm spatial resolution and 100 μs temporal resolution. In D-NBI, the bursting EPM is excited at slightly lower frequency and the IAE activity is nearly absent, likely due to an isotope effect and loss of wave-particle interaction. In these cases, neutral particle analysis shows little change in the core-localized high pitch fast ion content, and the fusion product profile indicates little change in the fast ion density profile, leaving unexplained the mechanism removing EPM drive. We measure a substantial redistribution of the fast ion profile due to strong lower-frequency ( 30kHz) MHD activity that accompanies the current profile relaxation in the RFP. Profile flattening is strongest in low bulk density discharges, which often occur with a total increase in global neutron flux from acceleration of the beam ions. Work supported by US DoE.

Monomeric tartrate resistant acid phosphatase induces insulin sensitive obesity.

Directory of Open Access Journals (Sweden)

Pernilla Lång

2008-03-01

Full Text Available Obesity is associated with macrophage infiltration of adipose tissue, which may link adipose inflammation to insulin resistance. However, the impact of inflammatory cells in the pathophysiology of obesity remains unclear. Tartrate resistant acid phosphatase (TRAP is an enzyme expressed by subsets of macrophages and osteoclasts that exists either as an enzymatically inactive monomer or as an active, proteolytically processed dimer.Using mice over expressing TRAP, we show that over-expression of monomeric, but not the dimeric form in adipose tissue leads to early onset spontaneous hyperplastic obesity i.e. many small fat cells. In vitro, recombinant monomeric, but not proteolytically processed TRAP induced proliferation and differentiation of mouse and human adipocyte precursor cells. In humans, monomeric TRAP was highly expressed in the adipose tissue of obese individuals. In both the mouse model and in the obese humans the source of TRAP in adipose tissue was macrophages. In addition, the obese TRAP over expressing mice exhibited signs of a low-grade inflammatory reaction in adipose tissue without evidence of abnormal adipocyte lipolysis, lipogenesis or insulin sensitivity.Monomeric TRAP, most likely secreted from adipose tissue macrophages, induces hyperplastic obesity with normal adipocyte lipid metabolism and insulin sensitivity.

Peracetic Acid Depolymerization of Biorefinery Lignin for Production of Selective Monomeric Phenolic Compounds

Energy Technology Data Exchange (ETDEWEB)

Ma, Ruoshui [Voiland School of Chemical Engineering and Bioengineering, Bioproducts, Science & Engineering Laboratory, Washington State University, 2710 Crimson Way Richland WA 99354 USA; Guo, Mond [Voiland School of Chemical Engineering and Bioengineering, Bioproducts, Science & Engineering Laboratory, Washington State University, 2710 Crimson Way Richland WA 99354 USA; Lin, Kuan-ting [Voiland School of Chemical Engineering and Bioengineering, Bioproducts, Science & Engineering Laboratory, Washington State University, 2710 Crimson Way Richland WA 99354 USA; Hebert, Vincent R. [Food and Environmental Laboratory, Washington State, University-TriCities, 2710 Crimson Way Richland WA 99354 USA; Zhang, Jinwen [Wood Materials and Engineering Laboratory, Washington State University, Pullman WA 99164 USA; Wolcott, Michael P. [Wood Materials and Engineering Laboratory, Washington State University, Pullman WA 99164 USA; Quintero, Melissa [Voiland School of Chemical Engineering and Bioengineering, Bioproducts, Science & Engineering Laboratory, Washington State University, 2710 Crimson Way Richland WA 99354 USA; Ramasamy, Karthikeyan K. [Chemical and Biological Process Development Group, Pacific Northwest National Laboratory, Richland WA 99354 USA; Chen, Xiaowen [National Bioenergy Center, National Renewable Energy Lab, 1617 Cole Blvd Golden CO 80127 USA; Zhang, Xiao [Voiland School of Chemical Engineering and Bioengineering, Bioproducts, Science & Engineering Laboratory, Washington State University, 2710 Crimson Way Richland WA 99354 USA

2016-07-04

Lignin is the largest source of renewable material with an aromatic skeleton. However, due to the recalcitrant and heterogeneous nature of the lignin polymer, it has been a challenge to effectively depolymerize lignin and produce high-value chemicals with high selectivity. In this study, a highly efficient lignin-to-monomeric phenolic compounds (MPC) conversion method based on peracetic acid (PAA) treatment was reported. PAA treatment of two biorefinery lignin samples, diluted acid pretreated corn stover lignin (DACSL) and steam exploded spruce lignin (SESPL), led to complete solubilization and production of selective hydroxylated monomeric phenolic compounds (MPC-H) and monomeric phenolic acid compounds (MPC-A) including 4-hydroxy-2-methoxyphenol, p-hydroxybenzoic acid, vanillic acid, syringic acid, and 3,4-dihydroxybenzoic acid. The maximized MPC yields obtained were 18 and 22 % based on the initial weight of the lignin in SESPL and DACSL, respectively. However, we found that the addition of niobium pentoxide catalyst to PAA treatment of lignin can significantly improve the MPC yields up to 47 %. The key reaction steps and main mechanisms involved in this new lignin-to-MPC valorization pathway were investigated and elucidated.

Peracetic Acid Depolymerization of Biorefinery Lignin for Production of Selective Monomeric Phenolic Compounds

Energy Technology Data Exchange (ETDEWEB)

Ma, Ruoshui [Voiland School of Chemical Engineering and Bioengineering, Bioproducts, Science & Engineering Laboratory, Washington State University, 2710 Crimson Way Richland WA 99354 USA; Guo, Mond [Voiland School of Chemical Engineering and Bioengineering, Bioproducts, Science & Engineering Laboratory, Washington State University, 2710 Crimson Way Richland WA 99354 USA; Lin, Kuan-ting [Voiland School of Chemical Engineering and Bioengineering, Bioproducts, Science & Engineering Laboratory, Washington State University, 2710 Crimson Way Richland WA 99354 USA; Hebert, Vincent R. [Food and Environmental Laboratory, Washington State, University-TriCities, 2710 Crimson Way Richland WA 99354 USA; Zhang, Jinwen [Wood Materials and Engineering Laboratory, Washington State University, Pullman WA 99164 USA; Wolcott, Michael P. [Wood Materials and Engineering Laboratory, Washington State University, Pullman WA 99164 USA; Quintero, Melissa [Voiland School of Chemical Engineering and Bioengineering, Bioproducts, Science & Engineering Laboratory, Washington State University, 2710 Crimson Way Richland WA 99354 USA; Ramasamy, Karthikeyan K. [Chemical and Biological Process Development Group, Pacific Northwest National Laboratory, Richland WA 99354 USA; Chen, Xiaowen [National Bioenergy Center, National Renewable Energy Lab, 1617 Cole Blvd Golden CO 80127 USA; Zhang, Xiao [Voiland School of Chemical Engineering and Bioengineering, Bioproducts, Science & Engineering Laboratory, Washington State University, 2710 Crimson Way Richland WA 99354 USA

2016-07-04

Lignin is the largest source of renewable material with an aromatic skeleton. However, due to the recalcitrant and heterogeneous nature of the lignin polymer as well as its complex side chain structures, it has been a challenge to effectively depolymerize lignin and produce high value chemicals with high selectivity. In this study, a highly efficient lignin-to-monomeric phenolic compounds (MPC) conversion method based on peracetic acid (PAA) treatment was reported. PAA treatment of two biorefinery lignin samples, diluted acid pretreated corn stover lignin (DACSL) and steam exploded spruce lignin (SESPL), led to complete solubilization and production of selective hydroxylated monomeric phenolic compounds (MPC-H) and monomeric phenolic acid compounds (MPC-A) inclduing 4-hydroxy-2-methoxyphenol, p-hydroxybenzoic acid, vanillic acid, syringic acid, and 3,4-dihydroxybenzoic acid. The maximized MPCs yields obtained were 18% and 22% based on the initial weight of the lignin in SESPL and DACSL respectively. However, we found that the addition of niobium pentoxide catalyst to PAA treatment of lignin can significantly improve the MPC yields up to 47%. The key reaction steps and main mechanisms involved in this new lignin-to-MPC valorization pathway were investigated and elucidated.

Distribution Dynamics of Recombinant Lactobacillus in the Gastrointestinal Tract of Neonatal Rats

Science.gov (United States)

Bao, Sujin; Zhu, Libin; Zhuang, Qiang; Wang, Lucia; Xu, Pin-Xian; Itoh, Keiji; Holzman, Ian R.; Lin, Jing

2013-01-01

One approach to deliver therapeutic agents, especially proteins, to the gastro-intestinal (GI) tract is to use commensal bacteria as a carrier. Genus Lactobacillus is an attractive candidate for use in this approach. However, a system for expressing exogenous proteins at a high level has been lacking in Lactobacillus. Moreover, it will be necessary to introduce the recombinant Lactobacillus into the GI tract, ideally by oral administration. Whether orally administered Lactobacillus can reach and reside in the GI tract has not been explored in neonates. In this study, we have examined these issues in neonatal rats. To achieve a high level of protein expression in Lactobacillus, we tested the impact of three promoters and two backbones on protein expression levels using mRFP1, a red fluorescent protein, as a reporter. We found that a combination of an L-lactate dehydrogenase (ldhL) promoter of Lactobacillus sakei with a backbone from pLEM415 yielded the highest level of reporter expression. When this construct was used to transform Lactobacillus casei, Lactobacillus delbrueckii and Lactobacillus acidophilus, high levels of mRFP1 were detected in all these species and colonies of transformed Lactobacillus appeared pink under visible light. To test whether orally administered Lactobacillus can be retained in the GI tract of neonates, we fed the recombinant Lactobacillus casei to neonatal rats. We found that about 3% of the bacteria were retained in the GI tract of the rats at 24 h after oral feeding with more recombinant Lactobacillus in the stomach and small intestine than in the cecum and colon. No mortality was observed throughout this study with Lactobacillus. In contrast, all neonatal rats died within 24 hours after fed with transformed E. coli. Taken together, our results indicate that Lactobacillus has the potential to be used as a vehicle for the delivery of therapeutic agents to neonates. PMID:23544119

Substrate-Induced Dimerization of Engineered Monomeric Variants of Triosephosphate Isomerase from Trichomonas vaginalis.

Directory of Open Access Journals (Sweden)

Samuel Lara-Gonzalez

Full Text Available The dimeric nature of triosephosphate isomerases (TIMs is maintained by an extensive surface area interface of more than 1600 Å2. TIMs from Trichomonas vaginalis (TvTIM are held in their dimeric state by two mechanisms: a ball and socket interaction of residue 45 of one subunit that fits into the hydrophobic pocket of the complementary subunit and by swapping of loop 3 between subunits. TvTIMs differ from other TIMs in their unfolding energetics. In TvTIMs the energy necessary to unfold a monomer is greater than the energy necessary to dissociate the dimer. Herein we found that the character of residue I45 controls the dimer-monomer equilibrium in TvTIMs. Unfolding experiments employing monomeric and dimeric mutants led us to conclude that dimeric TvTIMs unfold following a four state model denaturation process whereas monomeric TvTIMs follow a three state model. In contrast to other monomeric TIMs, monomeric variants of TvTIM1 are stable and unexpectedly one of them (I45A is only 29-fold less active than wild-type TvTIM1. The high enzymatic activity of monomeric TvTIMs contrast with the marginal catalytic activity of diverse monomeric TIMs variants. The stability of the monomeric variants of TvTIM1 and the use of cross-linking and analytical ultracentrifugation experiments permit us to understand the differences between the catalytic activities of TvTIMs and other marginally active monomeric TIMs. As TvTIMs do not unfold upon dimer dissociation, herein we found that the high enzymatic activity of monomeric TvTIM variants is explained by the formation of catalytic dimeric competent species assisted by substrate binding.

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)

First molecular identification of the transgene red fluorescent protein (RFP in transgenic ornamental zebrafish (Danio rerio introduced in Peru

Directory of Open Access Journals (Sweden)

Carlos Scotto

2013-09-01

Full Text Available In this paper the transgenic fluorescent red, orange and pink zebra fish (Danio rerio, found in local aquariums in Peru, were identified using the PCR technique to amplify the transgene RFP sea anemone belonging to Discosoma spp. The gene expression of the red fluorescent protein (RFP transgene was found to determine different gradients-of-bioluminescence (shades in color in each GMO fish analyzed. We performed sequence analysis of the two variants of the RFP along with six variants of the existing fluorescent protein GFP from the Genbank, this could help identify quickly if they are new genes or variants thereof as these novel fluorescent proteins may be introduced in aquatic GMO in the future. Thus, developing and improving biosecurity measures through its timely detection at the molecular genetic level.

SYNTHESIS AND STRUCTURE OF BIS(PHENYLTETRAMETHYLCYCLOPENTADIENYL)TITANIUM(III) HYDRIDE - THE FIRST MONOMERIC BIS(CYCLOPENTADIENYL)TITANIUM(III) HYDRIDE : The First Monomeric Bis(cyclopentadienyl)titanium(III) Hydride

NARCIS (Netherlands)

de Wolf, J.M.; Meetsma, A.; Teuben, J.H

1995-01-01

The first structurally characterized monomeric bis(cyclopentadienyl)titanium(III) hydride, (C(5)PhMe(4))(2)TiH (4), was synthesized by hydrogenolysis of (C(5)PhMe(4))(2)TiMe (5). Hydride 4 was found to be a monomeric bent sandwich by X-ray diffraction methods, and the pentamethylcyclopentadienyl

Peracetic Acid Depolymerization of Biorefinery Lignin for Production of Selective Monomeric Phenolic Compounds.

Science.gov (United States)

Ma, Ruoshui; Guo, Mond; Lin, Kuan-Ting; Hebert, Vincent R; Zhang, Jinwen; Wolcott, Michael P; Quintero, Melissa; Ramasamy, Karthikeyan K; Chen, Xiaowen; Zhang, Xiao

2016-07-25

Lignin is the largest source of renewable material with an aromatic skeleton. However, due to the recalcitrant and heterogeneous nature of the lignin polymer, it has been a challenge to effectively depolymerize lignin and produce high-value chemicals with high selectivity. In this study, a highly efficient lignin-to-monomeric phenolic compounds (MPC) conversion method based on peracetic acid (PAA) treatment was reported. PAA treatment of two biorefinery lignin samples, diluted acid pretreated corn stover lignin (DACSL) and steam exploded spruce lignin (SESPL), led to complete solubilization and production of selective hydroxylated monomeric phenolic compounds (MPC-H) and monomeric phenolic acid compounds (MPC-A) including 4-hydroxy-2-methoxyphenol, p-hydroxybenzoic acid, vanillic acid, syringic acid, and 3,4-dihydroxybenzoic acid. The maximized MPC yields obtained were 18 and 22 % based on the initial weight of the lignin in SESPL and DACSL, respectively. However, we found that the addition of niobium pentoxide catalyst to PAA treatment of lignin can significantly improve the MPC yields up to 47 %. The key reaction steps and main mechanisms involved in this new lignin-to-MPC valorization pathway were investigated and elucidated. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Quantification of the predominant monomeric catechins in baking chocolate standard reference material by LC/APCI-MS.

Science.gov (United States)

Nelson, Bryant C; Sharpless, Katherine E

2003-01-29

Catechins are polyphenolic plant compounds (flavonoids) that may offer significant health benefits to humans. These benefits stem largely from their anticarcinogenic, antioxidant, and antimutagenic properties. Recent epidemiological studies suggest that the consumption of flavonoid-containing foods is associated with reduced risk of cardiovascular disease. Chocolate is a natural cocoa bean-based product that reportedly contains high levels of monomeric, oligomeric, and polymeric catechins. We have applied solid-liquid extraction and liquid chromatography coupled with atmospheric pressure chemical ionization-mass spectrometry to the identification and determination of the predominant monomeric catechins, (+)-catechin and (-)-epicatechin, in a baking chocolate Standard Reference Material (NIST Standard Reference Material 2384). (+)-Catechin and (-)-epicatechin are detected and quantified in chocolate extracts on the basis of selected-ion monitoring of their protonated [M + H](+) molecular ions. Tryptophan methyl ester is used as an internal standard. The developed method has the capacity to accurately quantify as little as 0.1 microg/mL (0.01 mg of catechin/g of chocolate) of either catechin in chocolate extracts, and the method has additionally been used to certify (+)-catechin and (-)-epicatechin levels in the baking chocolate Standard Reference Material. This is the first reported use of liquid chromatography/mass spectrometry for the quantitative determination of monomeric catechins in chocolate and the only report certifying monomeric catechin levels in a food-based Standard Reference Material.

Tumour localization and pharmacokinetics of iodine-125 human monoclonal IgM antibody (COU-1) and its monomeric and half-monomeric fragments analysed in nude mice grafted with human tumour

International Nuclear Information System (INIS)

Ditzel, H.; Erb, K.; Rasmussen, J.W.; Jensenius, J.C.

1992-01-01

Human monoclonal IgM antibodies reactive with cancer-associated antigens may not have the optimal imaging capability due to their large size. Fragmentation of human IgM is less than straight-forward due to the loss of immunoreactivity. From the human monoclonal IgM antibody COU-1 we have prepared monomeric and half-monomeric fragments, which retain the ability to bind to colon cancer cells in vitro. The pharmacokinetics and tumour localization were evaluated in nude mice bearing human colon adenocarcinoma and human melanoma grafts. Faster clearance from the circulation was seen for the smaller half-monomeric fragment with a half-life (rapid phase/slow phase) of 2 h/16 h compared with the intact antibody, 4 h/25 h, and the monomeric fragment, 3 h/27 h. Intact COU-1 as well as the fragments accumulated in the colon tumour graft. Higher amounts of radioactivity were found in the colon tumour as compared to normal organs for intact COU-1 at days 4 and 6, for the monomeric fragment at day 4, and for the half-monomeric fragment at day 2 after injection. This investigation demonstrates the favourable biodistribution of the half monomeric COU-1 fragment. The fast clearance of this fragment resulted in a tumour-to-muscle ratio as high as 22 on day 2 after injection. Also, only this fragment gave a positive tumour-to-blood ratio. Normal IgM and its fragments were used as controls. Radioimmunoscintigraphy demonstrated the colon tumour discriminatory properties of each of the three iodine-labelled antibody preparations. The results compare favourably with previously reported investigations of the localization of human monoclonal antibodies and suggest that fragments of human monoclonal IgM antibodies may be useful tools for the immunodetection of cancer in patients. (orig.)

Effects of pulsed electric field on ULQ and RFP plasmas

International Nuclear Information System (INIS)

Watanabe, M.; Saito, K.; Suzuki, T.

1997-01-01

Dynamo activity and self-organization processes are investigated using the application of pulsed poloidal and toroidal electric fields on ULQ and RFP plasmas. Synchronized to the application of the pulsed electric fields, the remarkable responses of the several plasma parameters are observed. The plasma has a preferential magnetic field structure, and the external perturbation activates fluctuation to maintain the structure through dynamo effect. This process changes the total dissipation with the variation of magnetic helicity in the system, showing that self organization accompanies an enhanced dissipation. (author)

Experimental scaling of fluctuations and confinement with Lundquist number in the RFP

International Nuclear Information System (INIS)

Stoneking, M.R.; Chapman, J.T.; Prager, S.C.; Sarff, J.S.

1997-09-01

The scaling of the magnetic and velocity fluctuations with Lundquist number (S) is examined experimentally over a range of values from 7 x 10 4 to 10 6 in a reversed field pinch (RFP) plasma. Magnetic fluctuations do not scale uniquely with the Lundquist number. At high (relative) density, fluctuations scale as b∝S -0.18 , and fluctuations are almost independent of S at low relative density, b∝S -0.07 ; however both exponents fall in the range of theoretical and numerical predictions. At high relative density, the scaling of the energy confinement time follows expectations for transport in a stochastic magnetic field. A confinement scaling law (nτ E ∝β 4/5 T -7/10 A -3/5 I φ 2 ) is derived assuming the persistent dominance of stochastic magnetic diffusion in the RFP and on the measured scaling of magnetic fluctuations. The peak velocity fluctuations during a sawtooth cycle scale marginally stronger than magnetic fluctuations but weaker than a simple Ohm's law prediction. The sawtooth period is determined by a resistive-Alfvenic hybrid time (T saw ∝√(τ R τ Alf )) rather than a purely resistive time

Generation and Characterization of an IgG4 Monomeric Fc Platform.

Directory of Open Access Journals (Sweden)

Lu Shan

Full Text Available The immunoglobulin Fc region is a homodimer consisted of two sets of CH2 and CH3 domains and has been exploited to generate two-arm protein fusions with high expression yields, simplified purification processes and extended serum half-life. However, attempts to generate one-arm fusion proteins with monomeric Fc, with one set of CH2 and CH3 domains, are often plagued with challenges such as weakened binding to FcRn or partial monomer formation. Here, we demonstrate the generation of a stable IgG4 Fc monomer with a unique combination of mutations at the CH3-CH3 interface using rational design combined with in vitro evolution methodologies. In addition to size-exclusion chromatography and analytical ultracentrifugation, we used multi-angle light scattering (MALS to show that the engineered Fc monomer exhibits excellent monodispersity. Furthermore, crystal structure analysis (PDB ID: 5HVW reveals monomeric properties supported by disrupted interactions at the CH3-CH3 interface. Monomeric Fc fusions with Fab or scFv achieved FcRn binding and serum half-life comparable to wildtype IgG. These results demonstrate that this monomeric IgG4 Fc is a promising therapeutic platform to extend the serum half-life of proteins in a monovalent format.

Constitutive expression of fluorescent protein by Aspergillus var. niger and Aspergillus carbonarius to monitor fungal colonization in maize plants.

Science.gov (United States)

Palencia, Edwin Rene; Glenn, Anthony Elbie; Hinton, Dorothy Mae; Bacon, Charles Wilson

2013-09-01

Aspergillus niger and Aspergillus carbonarius are two species in the Aspergillus section Nigri (black-spored aspergilli) frequently associated with peanut (Arachis hypogea), maize (Zea mays), and other plants as pathogens. These infections are symptomless and as such are major concerns since some black aspergilli produce important mycotoxins, ochratoxins A, and the fumonisins. To facilitate the study of the black aspergilli-maize interactions with maize during the early stages of infections, we developed a method that used the enhanced yellow fluorescent protein (eYFP) and the monomeric red fluorescent protein (mRFP1) to transform A. niger and A. carbonarius, respectively. The results were constitutive expressions of the fluorescent genes that were stable in the cytoplasms of hyphae and conidia under natural environmental conditions. The hyphal in planta distribution in 21-day-old seedlings of maize were similar wild type and transformants of A. niger and A. carbonarius. The in planta studies indicated that both wild type and transformants internally colonized leaf, stem and root tissues of maize seedlings, without any visible disease symptoms. Yellow and red fluorescent strains were capable of invading epidermal cells of maize roots intercellularly within the first 3 days after inoculation, but intracellular hyphal growth was more evident after 7 days of inoculation. We also tested the capacity of fluorescent transformants to produce ochratoxin A and the results with A. carbonarius showed that this transgenic strain produced similar concentrations of this secondary metabolite. This is the first report on the in planta expression of fluorescent proteins that should be useful to study the internal plant colonization patterns of two ochratoxigenic species in the Aspergillus section Nigri. © 2013.

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

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

Pengaruh Mimosa Pada Penyamakan Kulit Jaket Domba Samak Nabati Menggunakan Sistem C-RFP, Ditinjau Dari Sifat Organoleptis, Fisis, Dan Morfologi Kulit

OpenAIRE

Sutyasmi, Sri; Widowati, Titik Purwati; Setyadewi, Noor Maryam

2016-01-01

The purpose of this study was to determine the effect of the use of mimosa, to manufacture environmentally friendly leather for jacket by using the C-RFP system (C=Conditioning, R=Rapid, F=Fass (drum), P=Powder), on physical, organoleptic, and morphology properties of leather. It was also to obtain a formula for vegetable tanning with C-RFP system. Pickled skins are conditioned (pre-tanning) using Sodotan TSN and Sodotan APR, and then Sodotan TSN was chosen due to it meets the requirements of...

Pengaruh mimosa pada penyamakan kulit jaket domba samak nabati menggunakan sistem C-RFP, ditinjau dari sifat organoleptis, fisis, dan morfologi kulit

OpenAIRE

Sri Sutyasmi; Titik Purwati Widowati; Noor Maryam Setyadewi

2016-01-01

The purpose of this study was to determine the effect of the use of mimosa, to manufacture environmentally friendly leather for jacket by using the C-RFP system (C=Conditioning, R=Rapid, F=Fass (drum), P=Powder), on physical, organoleptic, and morphology properties of leather. It was also to obtain a formula for vegetable tanning with C-RFP system. Pickled skins are conditioned (pre-tanning) using Sodotan TSN and Sodotan APR, and then Sodotan TSN was chosen due to it meets the requirements of...

Direct labelling of monomeric antibody fragments Fab' with 99mTc

International Nuclear Information System (INIS)

Li Jun; Wang Shizhen; Yang Ziyi

1994-01-01

Direct labelling method and conditions of monomeric antibody Fab' with 99m Tc were investigated. Polyclonal antibody IgG was digested with ficin to produce dimeric fragments F(ab') 2 , which was subsequently reduced to monomeric fragments Fab' with 2-mercaptoethylamine. Finally, Fab' was incubated with sodium gluconate (Sn(II)) kit solution and 99m TcO 4 - eluted at room temperature to form 99m Tc-Fab'. The labelling efficiency was 85%-95%. The stability of labelled products was satisfactory and the elimination rate was faster than 99m Tc-IgG

Induction of antibodies against epitopes inaccessible on the HIV type 1 envelope oligomer by immunization with recombinant monomeric glycoprotein 120

DEFF Research Database (Denmark)

Schønning, Kristian; Bolmstedt, A; Novotny, J

1998-01-01

An N-glycan (N306) at the base of the V3 loop of HIV-BRU gp120 is shielding a linear neutralization epitope at the tip of the V3 loop on oligomeric Env. In contrast, this epitope is readily antigenic on monomeric gp120. Immunization with recombinant monomeric HIV-BRU gp120 may thus be expected...... immunogenic structures inaccessible on the envelope oligomer. The limited ability of recombinant gp120 vaccines to induce neutralizing antibodies against primary isolates may thus not exclusively reflect genetic variation....

Monomeric insulins and their experimental and clinical implications.

Science.gov (United States)

Brange, J; Owens, D R; Kang, S; Vølund, A

1990-09-01

Due to the inherent pharmacokinetic properties of available insulins, normoglycemia is rarely, if ever, achieved in insulin-dependent diabetic patients without compromising their quality of life. Subcutaneous insulin absorption is influenced by many factors, among which the associated state of insulin (hexameric) in pharmaceutical formulation may be of importance. This review describes the development of a series of human insulin analogues with reduced tendency to self-association that, because of more rapid absorption, are better suited to meal-related therapy. DNA technology has made it possible to prepare insulins that remain dimeric or even monomeric at high concentration by introducing one or a few amino acid substitutions into human insulin. These analogues were characterized and used for elucidating the mechanisms involved in subcutaneous absorption and were investigated in preliminary clinical studies. Their relative receptor binding and in vitro potency (free-fat cell assay), ranging from 0.05 to 600% relative to human insulin, were strongly correlated (r = 0.97). In vivo, most of the analogues exhibited approximately 100% activity, explainable by a dominating receptor-mediated clearance. This was confirmed by clamp studies in which correlation between receptor binding and clearance was observed. Thus, an analogue with reduced binding and clearance gives higher circulating concentrations, counterbalancing the reduced potency at the cellular level. Absorption studies in pigs revealed a strong inverse correlation (r = 0.96) between the rate of subcutaneous absorption and the mean association state of the insulin analogues. These studies also demonstrated that monomeric insulins were absorbed three times faster than human insulin. In healthy subjects, rates of disappearance from subcutis were two to three times faster for dimeric and monomeric analogues than for human insulin. Concomitantly, a more rapid rise in plasma insulin concentration and an earlier

Development of bisphenol A-removing recombinant Escherichia coli by monomeric and dimeric surface display of bisphenol A-binding peptide.

Science.gov (United States)

Maruthamuthu, Murali Kannan; Hong, Jiyeon; Arulsamy, Kulandaisamy; Somasundaram, Sivachandiran; Hong, SoonHo; Choe, Woo-Seok; Yoo, Ik-Keun

2018-04-01

Peptide-displaying Escherichia coli cells were investigated for use in adsorptive removal of bisphenol A (BPA) both in Luria-Bertani medium including BPA or ATM thermal paper eluted wastewater. Two recombinant strains were constructed with monomeric and dimeric repeats of the 7-mer BPA-binding peptide (KSLENSY), respectively. Greater than threefold increased adsorption of BPA [230.4 µmol BPA per g dry cell weight (DCW)] was found in dimeric peptide-displaying cells compared to monomeric strains (63.4 µmol per g DCW) in 15 ppm BPA solution. The selective removal of BPA from a mixture of BPA analogs (bisphenol F and bisphenol S) was verified in both monomeric and dimeric peptide-displaying cells. The binding chemistry of BPA with the peptide was assumed, based on molecular docking analysis, to be the interaction of BPA with serine and asparagine residues within the 7-mer peptide sequence. The peptide-displaying cells also functioned efficiently in thermal paper eluted wastewater containing 14.5 ppm BPA.

Live imaging of intra- and extracellular pH in plants using pHusion, a novel genetically encoded biosensor

Science.gov (United States)

Gjetting, Kisten Sisse Krag; Ytting, Cecilie Karkov; Schulz, Alexander; Fuglsang, Anja Thoe

2012-01-01

Changes in pH are now widely accepted as a signalling mechanism in cells. In plants, proton pumps in the plasma membrane and tonoplast play a key role in regulation of intracellular pH homeostasis and maintenance of transmembrane proton gradients. Proton transport in response to external stimuli can be expected to be finely regulated spatially and temporally. With the ambition to follow such changes live, a new genetically encoded sensor, pHusion, has been developed. pHusion is especially designed for apoplastic pH measurements. It was constitutively expressed in Arabidopsis and targeted for expression in either the cytosol or the apoplast including intracellular compartments. pHusion consists of the tandem concatenation of enhanced green fluorescent protein (EGFP) and monomeric red fluorescent protein (mRFP1), and works as a ratiometric pH sensor. Live microscopy at high spatial and temporal resolution is highly dependent on appropriate immobilization of the specimen for microscopy. Medical adhesive often used in such experiments destroys cell viability in roots. Here a novel system for immobilizing Arabidopsis seedling roots for perfusion experiments is presented which does not impair cell viability. With appropriate immobilization, it was possible to follow changes of the apoplastic and cytosolic pH in mesophyll and root tissue. Rapid pH homeostasis upon external pH changes was reflected by negligible cytosolic pH fluctuations, while the apoplastic pH changed drastically. The great potential for analysing pH regulation in a whole-tissue, physiological context is demonstrated by the immediate alkalinization of the subepidermal apoplast upon external indole-3-acetic acid administration. This change is highly significant in the elongation zone compared with the root hair zone and control roots. PMID:22407646

Live imaging of intra- and extracellular pH in plants using pHusion, a novel genetically encoded biosensor.

Science.gov (United States)

Gjetting, Kisten Sisse Krag; Ytting, Cecilie Karkov; Schulz, Alexander; Fuglsang, Anja Thoe

2012-05-01

Changes in pH are now widely accepted as a signalling mechanism in cells. In plants, proton pumps in the plasma membrane and tonoplast play a key role in regulation of intracellular pH homeostasis and maintenance of transmembrane proton gradients. Proton transport in response to external stimuli can be expected to be finely regulated spatially and temporally. With the ambition to follow such changes live, a new genetically encoded sensor, pHusion, has been developed. pHusion is especially designed for apoplastic pH measurements. It was constitutively expressed in Arabidopsis and targeted for expression in either the cytosol or the apoplast including intracellular compartments. pHusion consists of the tandem concatenation of enhanced green fluorescent protein (EGFP) and monomeric red fluorescent protein (mRFP1), and works as a ratiometric pH sensor. Live microscopy at high spatial and temporal resolution is highly dependent on appropriate immobilization of the specimen for microscopy. Medical adhesive often used in such experiments destroys cell viability in roots. Here a novel system for immobilizing Arabidopsis seedling roots for perfusion experiments is presented which does not impair cell viability. With appropriate immobilization, it was possible to follow changes of the apoplastic and cytosolic pH in mesophyll and root tissue. Rapid pH homeostasis upon external pH changes was reflected by negligible cytosolic pH fluctuations, while the apoplastic pH changed drastically. The great potential for analysing pH regulation in a whole-tissue, physiological context is demonstrated by the immediate alkalinization of the subepidermal apoplast upon external indole-3-acetic acid administration. This change is highly significant in the elongation zone compared with the root hair zone and control roots.

Lipoamino acid-based micelles as promising delivery vehicles for monomeric amphotericin B.

Science.gov (United States)

Serafim, Cláudia; Ferreira, Inês; Rijo, Patrícia; Pinheiro, Lídia; Faustino, Célia; Calado, António; Garcia-Rio, Luis

2016-01-30

Lipoamino acid-based micelles have been developed as delivery vehicles for the hydrophobic drug amphotericin B (AmB). The micellar solubilisation of AmB by a gemini lipoamino acid (LAA) derived from cysteine and its equimolar mixtures with the bile salts sodium cholate (NaC) and sodium deoxycholate (NaDC), as well as the aggregation sate of the drug in the micellar systems, was studied under biomimetic conditions (phosphate buffered-saline, pH 7.4) using UV-vis spectroscopy. Pure surfactant systems and equimolar mixtures were characterized by tensiometry and important parameters were determined, such as critical micelle concentration (CMC), surface tension at the CMC (γCMC), maximum surface excess concentration (Γmax), and minimum area occupied per molecule at the water/air interface (Amin). Rheological behaviour from viscosity measurements at different shear rates was also addressed. Solubilisation capacity was quantified in terms of molar solubilisation ratio (χ), micelle-water partition coefficient (KM) and Gibbs energy of solubilisation (ΔGs°). Formulations of AmB in micellar media were compared in terms of drug loading, encapsulation efficiency, aggregation state of AmB and in vitro antifungal activity against Candida albicans. The LAA-containing micellar systems solubilise AmB in its monomeric and less toxic form and exhibit in vitro antifungal activity comparable to that of the commercial formulation Fungizone. Copyright © 2015 Elsevier B.V. All rights reserved.

Field error reduction experiment on the REPUTE-1 RFP device

International Nuclear Information System (INIS)

Toyama, H.; Shinohara, S.; Yamagishi, K.

1989-01-01

The vacuum chamber of the RFP device REPUTE-1 is a welded structure using 18 sets of 1 mm thick Inconel bellows (inner minor radius 22 cm) and 2.4 mm thick port segments arranged in toroidal geometry as shown in Fig. 1. The vacuum chamber is surrounded by 5 mm thick stainless steel shells. The time constant of the shell is 1 ms for vertical field penetration. The pulse length in REPUTE-1 is so far 3.2 ms (about 3 times longer than shell skin time). The port bypass plates have been attached as shown in Fig. 2 to reduce field errors so that the pulse length becomes longer and the loop voltage becomes lower. (author) 5 refs., 4 figs

Anthocyanins and Their Variation in Red Wines I. Monomeric Anthocyanins and Their Color Expression

OpenAIRE

Chang-Qing Duan; Malcolm J. Reeves; Qiu-Hong Pan; Lin Mu; Na-Na Liang; Fei He; Jun Wang

2012-01-01

Originating in the grapes, monomeric anthocyanins in young red wines contribute the majority of color and the supposed beneficial health effects related to their consumption, and as such they are recognized as one of the most important groups of phenolic metabolites in red wines. In recent years, our increasing knowledge of the chemical complexity of the monomeric anthocyanins, their stability, together with the phenomena such as self-association and copigmentation that can stabilize and enha...

Depression of voltage-activated Ca2+ release in skeletal muscle by activation of a voltage-sensing phosphatase.

Science.gov (United States)

Berthier, Christine; Kutchukian, Candice; Bouvard, Clément; Okamura, Yasushi; Jacquemond, Vincent

2015-04-01

Phosphoinositides act as signaling molecules in numerous cellular transduction processes, and phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2) regulates the function of several types of plasma membrane ion channels. We investigated the potential role of PtdIns(4,5)P2 in Ca(2+) homeostasis and excitation-contraction (E-C) coupling of mouse muscle fibers using in vivo expression of the voltage-sensing phosphatases (VSPs) Ciona intestinalis VSP (Ci-VSP) or Danio rerio VSP (Dr-VSP). Confocal images of enhanced green fluorescent protein-tagged Dr-VSP revealed a banded pattern consistent with VSP localization within the transverse tubule membrane. Rhod-2 Ca(2+) transients generated by 0.5-s-long voltage-clamp depolarizing pulses sufficient to elicit Ca(2+) release from the sarcoplasmic reticulum (SR) but below the range at which VSPs are activated were unaffected by the presence of the VSPs. However, in Ci-VSP-expressing fibers challenged by 5-s-long depolarizing pulses, the Ca(2+) level late in the pulse (3 s after initiation) was significantly lower at 120 mV than at 20 mV. Furthermore, Ci-VSP-expressing fibers showed a reversible depression of Ca(2+) release during trains, with the peak Ca(2+) transient being reduced by ∼30% after the application of 10 200-ms-long pulses to 100 mV. A similar depression was observed in Dr-VSP-expressing fibers. Cav1.1 Ca(2+) channel-mediated current was unaffected by Ci-VSP activation. In fibers expressing Ci-VSP and a pleckstrin homology domain fused with monomeric red fluorescent protein (PLCδ1PH-mRFP), depolarizing pulses elicited transient changes in mRFP fluorescence consistent with release of transverse tubule-bound PLCδ1PH domain into the cytosol; the voltage sensitivity of these changes was consistent with that of Ci-VSP activation, and recovery occurred with a time constant in the 10-s range. Our results indicate that the PtdIns(4,5)P2 level is tightly maintained in the transverse tubule membrane of the muscle fibers

Anthocyanins and Their Variation in Red Wines I. Monomeric Anthocyanins and Their Color Expression

Directory of Open Access Journals (Sweden)

Chang-Qing Duan

2012-02-01

Full Text Available Originating in the grapes, monomeric anthocyanins in young red wines contribute the majority of color and the supposed beneficial health effects related to their consumption, and as such they are recognized as one of the most important groups of phenolic metabolites in red wines. In recent years, our increasing knowledge of the chemical complexity of the monomeric anthocyanins, their stability, together with the phenomena such as self-association and copigmentation that can stabilize and enhance their color has helped to explain their color representation in red wine making and aging. A series of new enological practices were developed to improve the anthocyanin extraction, as well as their color expression and maintenance. This paper summarizes the most recent advances in the studies of the monomeric anthocyanins in red wines, emphasizing their origin, occurrence, color enhancing effects, their degradation and the effect of various enological practices on them.

Application of Modal Parameter Estimation Methods for Continuous Wavelet Transform-Based Damage Detection for Beam-Like Structures

Directory of Open Access Journals (Sweden)

Zhi Qiu

2015-02-01

Full Text Available This paper presents a hybrid damage detection method based on continuous wavelet transform (CWT and modal parameter identification techniques for beam-like structures. First, two kinds of mode shape estimation methods, herein referred to as the quadrature peaks picking (QPP and rational fraction polynomial (RFP methods, are used to identify the first four mode shapes of an intact beam-like structure based on the hammer/accelerometer modal experiment. The results are compared and validated using a numerical simulation with ABAQUS software. In order to determine the damage detection effectiveness between the QPP-based method and the RFP-based method when applying the CWT technique, the first two mode shapes calculated by the QPP and RFP methods are analyzed using CWT. The experiment, performed on different damage scenarios involving beam-like structures, shows that, due to the outstanding advantage of the denoising characteristic of the RFP-based (RFP-CWT technique, the RFP-CWT method gives a clearer indication of the damage location than the conventionally used QPP-based (QPP-CWT method. Finally, an overall evaluation of the damage detection is outlined, as the identification results suggest that the newly proposed RFP-CWT method is accurate and reliable in terms of detection of damage locations on beam-like structures.

Multistage modeling of protein dynamics with monomeric Myc oncoprotein as an example.

Science.gov (United States)

Liu, Jiaojiao; Dai, Jin; He, Jianfeng; Niemi, Antti J; Ilieva, Nevena

2017-03-01

We propose to combine a mean-field approach with all-atom molecular dynamics (MD) into a multistage algorithm that can model protein folding and dynamics over very long time periods yet with atomic-level precision. As an example, we investigate an isolated monomeric Myc oncoprotein that has been implicated in carcinomas including those in colon, breast, and lungs. Under physiological conditions a monomeric Myc is presumed to be an example of intrinsically disordered proteins that pose a serious challenge to existing modeling techniques. We argue that a room-temperature monomeric Myc is in a dynamical state, it oscillates between different conformations that we identify. For this we adopt the Cα backbone of Myc in a crystallographic heteromer as an initial ansatz for the monomeric structure. We construct a multisoliton of the pertinent Landau free energy to describe the Cα profile with ultrahigh precision. We use Glauber dynamics to resolve how the multisoliton responds to repeated increases and decreases in ambient temperature. We confirm that the initial structure is unstable in isolation. We reveal a highly degenerate ground-state landscape, an attractive set towards which Glauber dynamics converges in the limit of vanishing ambient temperature. We analyze the thermal stability of this Glauber attractor using room-temperature molecular dynamics. We identify and scrutinize a particularly stable subset in which the two helical segments of the original multisoliton align in parallel next to each other. During the MD time evolution of a representative structure from this subset, we observe intermittent quasiparticle oscillations along the C-terminal α helix, some of which resemble a translating Davydov's Amide-I soliton. We propose that the presence of oscillatory motion is in line with the expected intrinsically disordered character of Myc.

Factors Affecting Aerosol Radiative Forcing from Both Production-based and Consumption-based View

Science.gov (United States)

Wang, J.; Lin, J.; Ni, R.

2017-12-01

Aerosol radiative forcing (RF) is determined by emissions and various chemical-transport-radiative processes in the atmosphere, a multi-factor problem whose individual contributors have not been well quantified. This problem becomes more complicated when taking into account the role of international trade, which means reallocated aerosol RF due to separation of regions producing goods and emissions and regions consuming those goods. Here we analyze major factors affecting RF of secondary inorganic aerosols (SIOAs, including sulfate, nitrate and ammonium), primary organic aerosol (POA) and black carbon (BC), extending the work of Lin et al. (2016, Nature Geoscience). We contrast five factors determining production-based (RFp, due to a region's production of goods) and consumption-based (RFc, due to a region's consumption) forcing by 11 major regions, including population size, per capita output, emission intensity (emission per output), chemical efficiency (mass per unit emission) and radiative efficiency (RF per unit mass). Comparing across the 11 regions, East Asia produces the strongest RFp and RFc of SIOA and BC and the second largest RFp and RFc of POA primarily due to its high emission intensity. Although Middle East and North Africa has low emissions, its RFp is strengthened by its largest chemical efficiency for POA and BC and second largest chemical efficiency for SIOA. However, RFp of South-East Asia and Pacific is greatly weakened by its lowest chemical efficiency. Economic trade means that net importers (Western Europe, North America and Pacific OECD) have higher RFc than RFp by 50-100%. And such forcing difference is mainly due to the high emission intensity of the exporters supplying these regions. For North America, SIOA's RFc is 50% stronger than RFp, for that emission intensity of SIOA is 5.2 times in East Asia and 2.5 times in Latin America and Caribbean compared with that in North America, and the chemical efficiency in the top four exporters are

Rapid transcriptional pulsing dynamics of high expressing retroviral transgenes in embryonic stem cells.

Directory of Open Access Journals (Sweden)

Mandy Y M Lo

Full Text Available Single cell imaging studies suggest that transcription is not continuous and occurs as discrete pulses of gene activity. To study mechanisms by which retroviral transgenes can transcribe to high levels, we used the MS2 system to visualize transcriptional dynamics of high expressing proviral integration sites in embryonic stem (ES cells. We established two ES cell lines each bearing a single copy, self-inactivating retroviral vector with a strong ubiquitous human EF1α gene promoter directing expression of mRFP fused to an MS2-stem-loop array. Transfection of MS2-EGFP generated EGFP focal dots bound to the mRFP-MS2 stem loop mRNA. These transcription foci colocalized with the transgene integration site detected by immunoFISH. Live tracking of single cells for 20 minutes detected EGFP focal dots that displayed frequent and rapid fluctuations in transcription over periods as short as 25 seconds. Similarly rapid fluctuations were detected from focal doublet signals that colocalized with replicated proviral integration sites by immunoFISH, consistent with transcriptional pulses from sister chromatids. We concluded that retroviral transgenes experience rapid transcriptional pulses in clonal ES cell lines that exhibit high level expression. These events are directed by a constitutive housekeeping gene promoter and may provide precedence for rapid transcriptional pulsing at endogenous genes in mammalian stem cells.

Reducing and measuring fluctuations in the MST RFP: Enhancement of energy confinement and measurement of the MHD dynamo

International Nuclear Information System (INIS)

Den Hartog, D.J.; Almagri, A.F.

1996-09-01

A three- to five-fold enhancement of the energy confinement time in a reversed-field pinch (RFP) has been achieved in the Madison Symmetric Torus (MST) by reducing the amplitude of tearing mode fluctuations responsible for anomalous transport in the core of the RFP. By applying a transient poloidal inductive electric field to flatten the current density profile, the fluctuation amplitude b/B decreases from 1.5% to 0.8%, the electron temperature T e0 increases from 250 eV to 370 eV, the ohmic input power decreases from 4.5 MW to approximately 1.5 MW, the poloidal beta β 0 increases from 6% to 9%, and the energy confinement time τ E increases from 1 ms to ∼5 ms in I φ = 340 kA plasmas with density n = 1 x 10 19 m -3 . Current profile control methods are being developed for the RFP in a program to eliminate transport associated with these current-gradient-driven fluctuations. In addition to controlling the amplitude of the tearing modes, we are vigorously pursuing an understanding of the physics of these fluctuations. In particular, plasma flow, both equilibrium and fluctuating, plays a critical role in a diversity of physical phenomena in MST. The key results: 1) Edge probe measurements show that the MHD dynamo is active in low collisionality plasmas, while at high collisionality a new mechanism, the 'electron diamagnetic dynamo,' is observed. 2) Core spectroscopic measurements show that the toroidal velocity fluctuations of the plasma are coherent with the large-scale magnetic tearing modes; the scalar product of these two fluctuating quantities is similar to that expected for the MHD dynamo electromotive force. 3) Toroidal plasma flow in MST exhibits large radial shear and can be actively controlled, including unlocking locked discharges, by modifying E r with a robust biased probe. 24 refs

Iontophoresis of monomeric insulin analogues in vitro: effects of insulin charge and skin pretreatment.

Science.gov (United States)

Langkjaer, L; Brange, J; Grodsky, G M; Guy, R H

1998-01-23

The aim of this study was to investigate the influence of association state and net charge of human insulin analogues on the rate of iontophoretic transport across hairless mouse skin, and the effect of different skin pretreatments on said transport. No insulin flux was observed with anodal delivery probably because of degradation at the Ag/AgCl anode. The flux during cathodal iontophoresis through intact skin was insignificant for human hexameric insulin, and only low and variable fluxes were observed for monomeric insulins. Using stripped skin on the other hand, the fluxes of monomeric insulins with two extra negative charges were 50-100 times higher than that of hexameric human insulin. Introducing three additional charges led to a further 2-3-fold increase in flux. Wiping the skin gently with absolute alcohol prior to iontophoresis resulted in a 1000-fold increase in transdermal transport of insulin relative to that across untreated skin, i.e. to almost the same level as stripping the skin. The alcohol pretreatment reduced the electrical resistance of the skin, presumably by lipid extraction. In conclusion, monomeric insulin analogues with at least two extra negative charges can be iontophoretically delivered across hairless mouse skin, whereas insignificant flux is observed with human, hexameric insulin. Wiping the skin with absolute alcohol prior to iontophoresis gave substantially improved transdermal transport of monomeric insulins resulting in clinically relevant delivery rates for basal treatment.

Monomeric adiponectin modulates nitric oxide release and calcium movements in porcine aortic endothelial cells in normal/high glucose conditions.

Science.gov (United States)

Grossini, Elena; Farruggio, Serena; Qoqaiche, Fatima; Raina, Giulia; Camillo, Lara; Sigaudo, Lorenzo; Mary, David; Surico, Nicola; Surico, Daniela

2016-09-15

Perivascular adipose tissue can be involved in the process of cardiovascular pathology through the release of adipokines, namely adiponectins. Monomeric adiponectin has been shown to increase coronary blood flow in anesthetized pigs through increased nitric oxide (NO) release and the involvement of adiponectin receptor 1 (AdipoR1). The present study was therefore planned to examine the effects of monomeric adiponectin on NO release and Ca(2+) transients in porcine aortic endothelial cells (PAEs) in normal/high glucose conditions and the related mechanisms. PAEs were treated with monomeric adiponectin alone or in the presence of intracellular kinases blocker, AdipoR1 and Ca(2+)-ATPase pump inhibitors. The role of Na(+)/Ca(2+) exchanger was examined in experiments performed in zero Na(+) medium. NO release and intracellular Ca(2+) were measured through specific probes. In PAE cultured in normal glucose conditions, monomeric adiponectin elevated NO production and [Ca(2+)]c. Similar effects were observed in high glucose conditions, although the response was lower and not transient. The Ca(2+) mobilized by monomeric adiponectin originated from an intracellular pool thapsigargin- and ATP-sensitive and from the extracellular space. Moreover, the effects of monomeric adiponectin were prevented by kinase blockers and AdipoR1 inhibitor. Finally, in normal glucose condition, a role for Na(+)/Ca(2+) exchanger and Ca(2+)-ATPase pump in restoring Ca(2+) was found. Our results add new information about the control of endothelial function elicited by monomeric adiponectin, which would be achieved by modulation of NO release and Ca(2+) transients. A signalling related to Akt, ERK1/2 and p38MAPK downstream AdipoR1 would be involved. Copyright © 2016 Elsevier Inc. All rights reserved.

Monomeric adiponectin increases cell viability in porcine aortic endothelial cells cultured in normal and high glucose conditions: Data on kinases activation

Directory of Open Access Journals (Sweden)

Elena Grossini

2016-09-01

Full Text Available We found that monomeric adiponectin was able to increase cell viability in porcine aortic endothelial cells (PAE cultured both in normal and high glucose condition. Moreover, in normal glucose condition monomeric adiponectin increased p38MAPK, Akt, ERK1/2 and eNOS phosphorylation in a dose- and time-dependent way. Also in high glucose condition monomeric adiponectin increased eNOS and above kinases phosphorylation with similar patterns but at lower extent. For interpretation of the data presented in this article, please see the research article “Monomeric adiponectin modulates nitric oxide release and calcium movements in porcine aortic endothelial cells in normal/high glucose conditions” (Grossini et al., in press [1].

Expanding the genetic toolbox for Leptospira species by generation of fluorescent bacteria.

Science.gov (United States)

Aviat, Florence; Slamti, Leyla; Cerqueira, Gustavo M; Lourdault, Kristel; Picardeau, Mathieu

2010-12-01

Our knowledge of the genetics and molecular basis of the pathogenesis associated with Leptospira, in comparison to those of other bacterial species, is very limited. An improved understanding of pathogenic mechanisms requires reliable genetic tools for functional genetic analysis. Here, we report the expression of gfp and mRFP1 genes under the control of constitutive spirochetal promoters in both saprophytic and pathogenic Leptospira strains. We were able to reliably measure the fluorescence of Leptospira by fluorescence microscopy and a fluorometric microplate reader-based assay. We showed that the expression of the gfp gene had no significant effects on growth in vivo and pathogenicity in L. interrogans. We constructed an expression vector for L. biflexa that contains the lacI repressor, an inducible lac promoter, and gfp as the reporter, demonstrating that the lac system is functional in Leptospira. Green fluorescent protein (GFP) expression was induced by the addition of isopropyl-β-d-thiogalactopyranoside (IPTG) in L. biflexa transformants harboring the expression vector. Finally, we showed that GFP can be used as a reporter to assess promoter activity in different environmental conditions. These results may facilitate further advances for studying the genetics of Leptospira spp.

Natural monomeric form of fetal bovine serum acetylcholinesterase lacks the C-terminal tetramerization domain.

Science.gov (United States)

Saxena, Ashima; Hur, Regina S; Luo, Chunyuan; Doctor, Bhupendra P

2003-12-30

Acetylcholinesterase isolated from fetal bovine serum (FBS AChE) was previously characterized as a globular tetrameric form. Analysis of purified preparations of FBS AChE by gel permeation chromatography revealed the presence of a stable, catalytically active, monomeric form of this enzyme. The two forms could be distinguished from each other based on their molecular weight, hydrodynamic properties, kinetic properties, thermal stability, and the type of glycans they carry. No differences between the two forms were observed for the binding of classical inhibitors such as edrophonium and propidium or inhibitors that are current or potential drugs for the treatment of Alzheimer's disease such as (-) huperzine A and E2020; tacrine inhibited the monomeric form 2-3-fold more potently than the tetrameric form. Sequencing of peptides obtained from an in-gel tryptic digest of the monomer and tetramer by tandem mass spectrometry indicated that the tetramer consists of 583 amino acid residues corresponding to the mature form of the enzyme, whereas the monomer consists of 543-547 amino acid residues. The subunit molecular weight of the protein component of the monomer (major species) was determined to be 59 414 Da and that of the tetramer as 64 239 Da. The N-terminal of the monomer and the tetramer was Glu, suggesting that the monomer is not a result of truncation at the N-terminal. The only differences detected were at the C-terminus. The tetramer yielded the expected C-terminus, CSDL, whereas the C-terminus of the monomer yielded a mixture of peptides, of which LLSATDTLD was the most abundant. These results suggest that monomeric FBS AChE is trimmed at the C-terminus, and the results are consistent with the involvement of C-terminal amino acids in the assembly of monomers into tetramers.

Distinct subcellular trafficking resulting from monomeric vs multimeric targeting to endothelial ICAM-1: implications for drug delivery.

Science.gov (United States)

Ghaffarian, Rasa; Muro, Silvia

2014-12-01

Ligand-targeted, receptor-mediated endocytosis is commonly exploited for intracellular drug delivery. However, cells-surface receptors may follow distinct endocytic fates when bound by monomeric vs multimeric ligands. Our purpose was to study this paradigm using ICAM-1, an endothelial receptor involved in inflammation, to better understand its regulation and potential for drug delivery. Our procedure involved fluorescence microscopy of human endothelial cells to determine the endocytic behavior of unbound ICAM-1 vs ICAM-1 bound by model ligands: monomeric (anti-ICAM) vs multimeric (anti-ICAM biotin-streptavidin conjugates or anti-ICAM coated onto 100 nm nanocarriers). Our findings suggest that both monomeric and multimeric ligands undergo a similar endocytic pathway sensitive to amiloride (∼50% inhibition), but not inhibitors of clathrin-pits or caveoli. After 30 min, ∼60-70% of both ligands colocalized with Rab11a-compartments. By 3-5 h, ∼65-80% of multimeric anti-ICAM colocalized with perinuclear lysosomes with ∼60-80% degradation, while 70% of monomeric anti-ICAM remained associated with Rab11a at the cell periphery and recycled to and from the cell-surface with minimal (drug delivery.

Steady-state acceptor fluorescence anisotropy imaging under evanescent excitation for visualisation of FRET at the plasma membrane.

Directory of Open Access Journals (Sweden)

Viviane Devauges

Full Text Available We present a novel imaging system combining total internal reflection fluorescence (TIRF microscopy with measurement of steady-state acceptor fluorescence anisotropy in order to perform live cell Förster Resonance Energy Transfer (FRET imaging at the plasma membrane. We compare directly the imaging performance of fluorescence anisotropy resolved TIRF with epifluorescence illumination. The use of high numerical aperture objective for TIRF required correction for induced depolarization factors. This arrangement enabled visualisation of conformational changes of a Raichu-Cdc42 FRET biosensor by measurement of intramolecular FRET between eGFP and mRFP1. Higher activity of the probe was found at the cell plasma membrane compared to intracellularly. Imaging fluorescence anisotropy in TIRF allowed clear differentiation of the Raichu-Cdc42 biosensor from negative control mutants. Finally, inhibition of Cdc42 was imaged dynamically in live cells, where we show temporal changes of the activity of the Raichu-Cdc42 biosensor.

Pellet injection in the RFP (Reversed Field Pinch)

Science.gov (United States)

Wurden, G. A.; Weber, P. G.; Munson, C. P.; Cayton, T. E.; Bunting, C. A.; Carolan, P. G.

Observation of pellets injected into the ZT-40M Reversed Field Pinch has allowed a new twist on the usual tokamak ablation physics modeling. The RFP provides a strong ohmic heating regime with relatively high electron drift parameter (xi sub drift approx. 0.2), in the presence of a highly sheared magnetic field geometry. In situ photos of the pellet ablation cloud using a grated-intensified CCD camera, as well as two-view integrated photos of the pellet trajectory show substantial modification of the original pellet trajectory, in both direction and speed. Depending on the launch geometry, increases in the initial 500 m/s pellet speed by 50 percent were observed, and a ski jump deflector plate in the launch port has been used to counteract strong poloidal curvature. In contrast to the tokamak, the D sub alpha light signature is strongest near the edge, and weaker in the plasma center. Additional information on ion temperature response to pellet injection with 20 microsec time resolution has been obtained using a 5-channel neutral particle analyzer (NPA). The energy confinement is transiently degraded while the beta is largely unchanged. This may be indicative of pellet injection into a high-beta plasma operating at fixed beta.

Pellet injection in the RFP [Reversed Field Pinch

International Nuclear Information System (INIS)

Wurden, G.A.; Weber, P.G.; Munson, C.P.; Cayton, T.E.; Bunting, C.A.; Carolan, P.G.

1988-01-01

Observation of pellets injected into the ZT-40M Reversed Field Pinch has allowed a new twist on the usual tokamak ablation physics modeling. The RFP provides a strong ohmic heating regime with relatively high electron drift parameter (ξ/sub drift/ /approximately/ 0.2), in the presence of a highly sheared magnetic field geometry. In situ photos of the pellet ablation cloud using a grated-intensified CCD camera, as well as two-view integrated photos of the pellet trajectory show substantial modification of the original pellet trajectory, in both direction and speed. Depending on the launch geometry, increases in the initial 500 m/s pellet speed by 50% have been observed, and a ski jump deflector plate in the launch port has been used to counteract strong poloidal curvature. In contrast to the tokamak, the D/sub α/ light signature is strongest near the edge, and weaker in the plasma center. Additional information on ion temperature response to pellet injection with 20 μsec time resolution has been obtained using a 5-channel neutral particle analyzer (NPA). The energy confinement is transiently degraded while the beta is largely unchanged. This may be indicative of pellet injection into a high-beta plasma operating at fixed beta. 10 refs., 6 figs

Heterogeneous transgene expression in the retinas of the TH-RFP, TH-Cre, TH-BAC-Cre and DAT-Cre mouse lines.

Science.gov (United States)

Vuong, H E; Pérez de Sevilla Müller, L; Hardi, C N; McMahon, D G; Brecha, N C

2015-10-29

Transgenic mouse lines are essential tools for understanding the connectivity, physiology and function of neuronal circuits, including those in the retina. This report compares transgene expression in the retina of a tyrosine hydroxylase (TH)-red fluorescent protein (RFP) mouse line with three catecholamine-related Cre recombinase mouse lines [TH-bacterial artificial chromosome (BAC)-, TH-, and dopamine transporter (DAT)-Cre] that were crossed with a ROSA26-tdTomato reporter line. Retinas were evaluated and immunostained with commonly used antibodies including those directed to TH, GABA and glycine to characterize the RFP or tdTomato fluorescent-labeled amacrine cells, and an antibody directed to RNA-binding protein with multiple splicing to identify ganglion cells. In TH-RFP retinas, types 1 and 2 dopamine (DA) amacrine cells were identified by their characteristic cellular morphology and type 1 DA cells by their expression of TH immunoreactivity. In the TH-BAC-, TH-, and DAT-tdTomato retinas, less than 1%, ∼ 6%, and 0%, respectively, of the fluorescent cells were the expected type 1 DA amacrine cells. Instead, in the TH-BAC-tdTomato retinas, fluorescently labeled AII amacrine cells were predominant, with some medium diameter ganglion cells. In TH-tdTomato retinas, fluorescence was in multiple neurochemical amacrine cell types, including four types of polyaxonal amacrine cells. In DAT-tdTomato retinas, fluorescence was in GABA immunoreactive amacrine cells, including two types of bistratified and two types of monostratified amacrine cells. Although each of the Cre lines was generated with the intent to specifically label DA cells, our findings show a cellular diversity in Cre expression in the adult retina and indicate the importance of careful characterization of transgene labeling patterns. These mouse lines with their distinctive cellular labeling patterns will be useful tools for future studies of retinal function and visual processing. Published by Elsevier

Stability of human interferon-beta 1: oligomeric human interferon-beta 1 is inactive but is reactivated by monomerization.

Science.gov (United States)

Utsumi, J; Yamazaki, S; Kawaguchi, K; Kimura, S; Shimizu, H

1989-10-05

Human interferon-beta 1 is extremely stable is a low ionic strength solution of pH 2 such as 10 mM HCl at 37 degrees C. However, the presence of 0.15 M NaCl led to a remarkable loss of antiviral activity. The molecular-sieve high-performance liquid chromatography revealed that, whereas completely active human interferon-beta 1 eluted as a 25 kDa species (monomeric form), the inactivated preparation eluted primarily as a 90 kDa species (oligomeric form). The specific activity (units per mg protein) of the oligomeric form was approx. 10% of that of the monomeric form. This observation shows that oligomeric human interferon-beta 1 is apparently in an inactive form. When the oligomeric eluate was resolved by polyacrylamide gel containing sodium dodecyl sulphate (SDS), it appeared to be monomeric under non-reducing conditions. Monomerization of the oligomeric human interferon-beta 1 by treatment with 1% SDS, fully regenerated its antiviral activity. These results suggest that the inactivation of the human interferon-beta 1 preparation was caused by its oligomerization via hydrophobic interactions without the formation of intermolecular disulphide bonds. These oligomers can be dissociated by SDS to restore biological activity.

Principal Component Regression Analysis of the Relation Between CIELAB Color and Monomeric Anthocyanins in Young Cabernet Sauvignon Wines

Directory of Open Access Journals (Sweden)

Chang-Qing Duan

2008-11-01

Full Text Available Color is one of the key characteristics used to evaluate the sensory quality of red wine, and anthocyanins are the main contributors to color. Monomeric anthocyanins and CIELAB color values were investigated by HPLC-MS and spectrophotometry during fermentation of Cabernet Sauvignon red wine, and principal component regression (PCR, a statistical tool, was used to establish a linkage between the detected anthocyanins and wine coloring. The results showed that 14 monomeric anthocyanins could be identified in wine samples, and all of these anthocyanins were negatively correlated with the L*, b* and H*ab values, but positively correlated with a* and C*ab values. On an equal concentration basis for each detected anthocyanin, cyanidin-3-O-glucoside (Cy3-glu had the most influence on CIELAB color value, while malvidin 3-O-glucoside (Mv3-glu had the least. The color values of various monomeric anthocyanins were influenced by their structures, substituents on the B-ring, acyl groups on the glucoside and the molecular steric structure. This work develops a statistical method for evaluating correlation between wine color and monomeric anthocyanins, and also provides a basis for elucidating the effect of intramolecular copigmentation on wine coloring.

Discovery of Radioiodinated Monomeric Anthraquinones as a Novel Class of Necrosis Avid Agents for Early Imaging of Necrotic Myocardium.

Science.gov (United States)

Wang, Qin; Yang, Shengwei; Jiang, Cuihua; Li, Jindian; Wang, Cong; Chen, Linwei; Jin, Qiaomei; Song, Shaoli; Feng, Yuanbo; Ni, Yicheng; Zhang, Jian; Yin, Zhiqi

2016-02-16

Assessment of myocardial viability is deemed necessary to aid in clinical decision making whether to recommend revascularization therapy for patients with myocardial infarction (MI). Dianthraquinones such as hypericin (Hyp) selectively accumulate in necrotic myocardium, but were unsuitable for early imaging after administration to assess myocardial viability. Since dianthraquinones can be composed by coupling two molecules of monomeric anthraquinone and the active center can be found by splitting chemical structure, we propose that monomeric anthraquinones may be effective functional groups for necrosis targetability. In this study, eight radioiodinated monomeric anthraquinones were evaluated as novel necrosis avid agents (NAAs) for imaging of necrotic myocardium. All (131)I-anthraquinones showed high affinity to necrotic tissues and (131)I-rhein emerged as the most promising compound. Infarcts were visualized on SPECT/CT images at 6 h after injection of (131)I-rhein, which was earlier than that with (131)I-Hyp. Moreover, (131)I-rhein showed satisfactory heart-to-blood, heart-to-liver and heart-to-lung ratios for obtaining images of good diagnostic quality. (131)I-rhein was a more promising "hot spot imaging" tracer for earlier visualization of necrotic myocardium than (131)I-Hyp, which supported further development of radiopharmaceuticals based on rhein for SPECT/CT ((123)I and (99m)Tc) or PET/CT imaging ((18)F and (124)I) of myocardial necrosis.

Management of root rot and root knot disease of mungbean with the application of mycorrhizospheric fluorescent pseudomonas under field condition

International Nuclear Information System (INIS)

Bokhari, S.S.; Tariq, S.; Ali, S.A.

2014-01-01

The mycorrhizosphere is the region around a mycorrhizal fungus in which nutrients released from the hyphae increases microbial population and its activities. In this study five mycorrhizospheric fluorescent Pseudomonas (MRFP) were evaluated for biocontrol potential under field condition using mungbean (Vigna radiata) as test plant. MRFP-249 significantly reduced Fusarium solani, Rhizoctonia solani and Macrophomina phaseolina. Whereas MRFP246 and MRFP-247 were also found effective against M. phaseolina. Mycorrhizospheric fluorescent Pseudomonas were also found effective against root knot nematode by reducing the galls on roots and nematode's penetration in roots. Highest fresh shoot weight and plant height was produced by MRFP-248. Plants grown in soil treated with Pseudomonas showed higher number of VAM spores around the mungbean roots than untreated control plants. The mycorrhizal symbiosis should not be considered merely as bipartite, plant-fungus interaction, but should instead include the associated microorganisms, particularly fluorescent Pseudomonas. (author)

Monomeric insulins obtained by protein engineering and their medical implications.

Science.gov (United States)

Brange, J; Ribel, U; Hansen, J F; Dodson, G; Hansen, M T; Havelund, S; Melberg, S G; Norris, F; Norris, K; Snel, L

1988-06-16

The use of insulin as an injected therapeutic agent for the treatment of diabetes has been one of the outstanding successes of modern medicine. The therapy has, however, had its associated problems, not least because injection of insulin does not lead to normal diurnal concentrations of insulin in the blood. This is especially true at meal times when absorption from subcutaneous tissue is too slow to mimic the normal rapid increments of insulin in the blood. In the neutral solutions used for therapy, insulin is mostly assembled as zinc-containing hexamers and this self-association, which under normal physiological circumstances functions to facilitate proinsulin transport, conversion and intracellular storage, may limit the rate of absorption. We now report that it is possible, by single amino-acid substitutions, to make insulins which are essentially monomeric at pharmaceutical concentrations (0.6 mM) and which have largely preserved their biological activity. These monomeric insulins are absorbed two to three times faster after subcutaneous injection than the present rapid-acting insulins. They are therefore capable of giving diabetic patients a more physiological plasma insulin profile at the time of meal consumption.

Confinement studies of a high current density RFP in the Extrap T1 Upgrade device

International Nuclear Information System (INIS)

Drake, J.R.; Brzozowski, J.H.; Brunsell, P.; Hellblom, G.; Karlsson, P.; Mazur, S.; Nordlund, P.; Welander, A.; Zastrow, K.D.

1992-01-01

Confinement studies have been carried out on the Extrap T1 device operated in the reversed field pinch (RFP) mode. Extrap T1 is a small device with a major radius of R=0.5 m and a high aspect ratio, R/a=8.9. For these experiments, the device has been operated with a resistive shell with measured, toroidally-averaged flux penetration times of τ sv = 500μs (vertical) and τ sR =300μs (radial). The pulse lengths are about 600 μs, which is slightly longer than the shell penetration time. The purpose of these experiments is to study energy confinement in a high aspect-ratio, high current-density RFP device with a resistive shell. The device can be operated with high current densities which exceed 20 MAm -2 on axis. For these discharges, the average electron density is relatively high, ≅ 1x10 20 m -3 . Therefore, although the average current density exceeds 5 MAm -2 , the important parameter / ≅ I/N is maintained less than 1x10 -13 Am, where N is the line density. The plasma diagnostics for the device include a single chord CO 2 laser interferometer ( ), single point Thomson scattering (T e , n o ), VUV and visible spectroscopy (T e , Z eff ) surface barrier diodes for soft X-ray measurements (T e ), bolometry (P rad ), surface probes (Γ p ,T i ) and comprehensive magnetic diagnostics for both equilibrium and magnetic fluctuation studies. (author) 5 refs., 1 fig., 1 tab

Mapping monomeric threading to protein-protein structure prediction.

Science.gov (United States)

Guerler, Aysam; Govindarajoo, Brandon; Zhang, Yang

2013-03-25

The key step of template-based protein-protein structure prediction is the recognition of complexes from experimental structure libraries that have similar quaternary fold. Maintaining two monomer and dimer structure libraries is however laborious, and inappropriate library construction can degrade template recognition coverage. We propose a novel strategy SPRING to identify complexes by mapping monomeric threading alignments to protein-protein interactions based on the original oligomer entries in the PDB, which does not rely on library construction and increases the efficiency and quality of complex template recognitions. SPRING is tested on 1838 nonhomologous protein complexes which can recognize correct quaternary template structures with a TM score >0.5 in 1115 cases after excluding homologous proteins. The average TM score of the first model is 60% and 17% higher than that by HHsearch and COTH, respectively, while the number of targets with an interface RMSD benchmark proteins. Although the relative performance of SPRING and ZDOCK depends on the level of homology filters, a combination of the two methods can result in a significantly higher model quality than ZDOCK at all homology thresholds. These data demonstrate a new efficient approach to quaternary structure recognition that is ready to use for genome-scale modeling of protein-protein interactions due to the high speed and accuracy.

Improved detection of a staphylococcal infection by monomeric and protein A-purified polyclonal human immunoglobulin

International Nuclear Information System (INIS)

Calame, W.

1993-01-01

The present study was undertaken to compare the technetium-99m labelled non-specific polyclonal human immunoglobulin (Ig) with 99m Tc-labelled monomeric human immunoglobulin (m-Ig), 99m Tc-labelled, protein A-purified, human immunoglobulin (A-IG) and 99m Tc-labelled monomeric, protein A-purified, human immunoglobulin (mA-Ig) as tracer agents for the detection of a thigh infection with Staphylococcus aureus. In vitro the binding of the various tracer agents to bacteria at various intervals was determined. For the in vivo evaluation, mice were infected and received one of the various labelled proteins. Scintigrams were made 0.25, 1, 4 and 24 h later. All 99m Tc-labelled Igs bound to bacteria in vitro: The percentages of binding for the m-Ig (from 1 h onwards) and A-Ig and mA-Ig (from 3 h onwards) were significantly higher than that for Ig. The in vivo target-to-non-target (T/NT) ratios were significantly higher from 4 h onwards for all purified Igs than for Ig. Protein A-purified Ig yielded higher T/NT ratios than m-Ig. Furthermore, the amount of activity in the liver was significantly lower 24 h after administration of m-Ig, A-Ig and mA-Ig than after administration of Ig. It is concluded that in this experimental infection 99m Tc-labelled monomeric Ig localizes a staphylococcal thigh infection better and faster than 99m Tc-labelled unpurified Ig. However, the accumulation obtained with protein A-purified Ig or protein A-purified monomeric Ig was the highest of all tracer agents tested. (orig.)

Characterization of monomeric intermediates during VSV glycoprotein structural transition.

Directory of Open Access Journals (Sweden)

Aurélie A Albertini

2012-02-01

Full Text Available Entry of enveloped viruses requires fusion of viral and cellular membranes, driven by conformational changes of viral glycoproteins. Crystal structures provide static pictures of pre- and post-fusion conformations of these proteins but the transition pathway remains elusive. Here, using several biophysical techniques, including analytical ultracentrifugation, circular dichroïsm, electron microscopy and small angle X-ray scattering, we have characterized the low-pH-induced fusogenic structural transition of a soluble form of vesicular stomatitis virus (VSV glycoprotein G ectodomain (G(th, aa residues 1-422, the fragment that was previously crystallized. While the post-fusion trimer is the major species detected at low pH, the pre-fusion trimer is not detected in solution. Rather, at high pH, G(th is a flexible monomer that explores a large conformational space. The monomeric population exhibits a marked pH-dependence and adopts more elongated conformations when pH decreases. Furthermore, large relative movements of domains are detected in absence of significant secondary structure modification. Solution studies are complemented by electron micrographs of negatively stained viral particles in which monomeric ectodomains of G are observed at the viral surface at both pH 7.5 and pH 6.7. We propose that the monomers are intermediates during the conformational change and thus that VSV G trimers dissociate at the viral surface during the structural transition.

Holistic versus monomeric strategies for hydrological modelling of human-modified hydrosystems

Directory of Open Access Journals (Sweden)

I. Nalbantis

2011-03-01

Full Text Available The modelling of human-modified basins that are inadequately measured constitutes a challenge for hydrological science. Often, models for such systems are detailed and hydraulics-based for only one part of the system while for other parts oversimplified models or rough assumptions are used. This is typically a bottom-up approach, which seeks to exploit knowledge of hydrological processes at the micro-scale at some components of the system. Also, it is a monomeric approach in two ways: first, essential interactions among system components may be poorly represented or even omitted; second, differences in the level of detail of process representation can lead to uncontrolled errors. Additionally, the calibration procedure merely accounts for the reproduction of the observed responses using typical fitting criteria. The paper aims to raise some critical issues, regarding the entire modelling approach for such hydrosystems. For this, two alternative modelling strategies are examined that reflect two modelling approaches or philosophies: a dominant bottom-up approach, which is also monomeric and, very often, based on output information, and a top-down and holistic approach based on generalized information. Critical options are examined, which codify the differences between the two strategies: the representation of surface, groundwater and water management processes, the schematization and parameterization concepts and the parameter estimation methodology. The first strategy is based on stand-alone models for surface and groundwater processes and for water management, which are employed sequentially. For each model, a different (detailed or coarse parameterization is used, which is dictated by the hydrosystem schematization. The second strategy involves model integration for all processes, parsimonious parameterization and hybrid manual-automatic parameter optimization based on multiple objectives. A test case is examined in a hydrosystem in Greece

Primera identificación molecular del transgen de la proteína fluorescente roja (RFP en peces Cebra (Danio rerio transgénicos ornamentales introducidos en el Perú

Directory of Open Access Journals (Sweden)

Carlos Scotto

2013-01-01

Full Text Available En el presente trabajo se identificó por primera vez peces Cebra transgénicos (Danio rerio fluorescentes de color rojo, naranja y rosado introducidos al territorio peruano de acuarios locales utilizando la técnica de PCR para amplificar el transgen RFP perteneciente a la anémona marina Discosoma spp. Se encontró una expresión génica diferencial del transgen de la proteína fluorescente roja (RFP que determinaría una gradiente de bioluminiscencia para cada color entre los peces OVM analizados. Se realizó un análisis de secuencias de las dos variantes de la RFP junto con las seis variantes de la GFP de proteínas fluorescentes existentes en el Genbank que podrían ayudar a identificar rápidamente si son nuevos genes o si son nuevas variantes de éstas proteínas fluorescentes y que podrían ser utilizadas en otros OVMs hidrobiológicos a futuro. De este modo, desarrollar y optimizar las medidas de bioseguridad mediante su oportuna detección a nivel genético molecular.

Preparation of a highly concentrated, completely monomeric, active sarcoplasmic reticulum Ca2+-ATPase.

Science.gov (United States)

Lüdi, H; Hasselbach, W

1985-11-21

Sarcoplasmic reticulum vesicles from fast skeletal muscle were partially delipidated with sodium cholate at high ionic strength and sedimented in a discontinuous sucrose gradient. Phospholipid content was reduced from 0.777 mumol/mg protein to 0.242 mumol/mg protein. As judged from gel electrophoresis and high pressure liquid gel chromatography, accessory proteins were removed during centrifugation and the Ca2+-ATPase was obtained in an almost pure form. Addition of myristoylglycerophosphocholine (1 mg/mg protein) reactivates ATPase and dinitrophenylphosphatase activity to the same degree obtained with native vesicles. Using the analytical ultracentrifuge it could be demonstrated that the reactivated Ca2+-ATPase was present exclusively in a monomeric state. These results were obtained at high and low ionic strength and up to a protein concentration of 10 mg/ml. Therefore this preparation should be very useful to investigate differences between oligomeric and monomeric Ca2+-ATPase.

11 Efficacy and Tolerability of HDM Injective Immunotherapy With Monomeric Allergoid

Science.gov (United States)

Compalati, Enrico; Atzeni, Isabella; Cabras, Sergio; Fancello, Paolo; Gaspardini, Giulio; Longo, Rocco; Patella, Vincenzo; Tore, Giorgio

2012-01-01

Background Subcutaneous immunotherapy (SCIT) is an effective treatment of respiratory allergy and carbamylated monomeric allergoids (monoids), by virtue of their reduced IgE-binding activity, resulted clinically safe by sublingual administration. Purpose of this study was to investigate the efficacy and tolerability of immunotherapy with house dust mites (HDM) monoid administered by injective route in patients with allergic rhinoconjunctivitis (AR). Methods A preparation of 0.70 mL of 10 BU/mL containing modified extract with 50% Dermatophagoides pteronyssinus and 50% Dermatophagoides farinae (amount of major allergen: 4 μg of group 1 per milliliter) was delivered monthly for 12 months, following a 5-week build-up induction phase (0.10–0.20–0.30–0.50–0.70 mL), to 58 patients (60% males, mean age 25.1 ± 12.7) suffering from AR due to mites for at least 2 years, whereas 60 patients with similar baseline characteristics were observed as controls. All patients were allowed to assume traditional drug therapy for their condition. At the end of the study changes from baseline in symptoms scores, in number of days with drug assumption, in severity of AR (according to ARIA classification) were compared between the 2 groups; moreover an overall assessment of clinical efficacy and tolerability was based on patients' and physicians' judgements (unsatisfactory, mild, good, optimal). Results In respect to baseline both groups showed, after 1 year, an improvement in symptoms score (P allergoid was associated with a significant clinical benefit observed through objective and subjective outcomes; the traditional safety of monomeric allergoids was confirmed by the subjective judgements of tolerability.

Metal-free ALS variants of dimeric human Cu,Zn-superoxide dismutase have enhanced populations of monomeric species.

Directory of Open Access Journals (Sweden)

Anna-Karin E Svensson

2010-04-01

Full Text Available Amino acid replacements at dozens of positions in the dimeric protein human, Cu,Zn superoxide dismutase (SOD1 can cause amyotrophic lateral sclerosis (ALS. Although it has long been hypothesized that these mutations might enhance the populations of marginally-stable aggregation-prone species responsible for cellular toxicity, there has been little quantitative evidence to support this notion. Perturbations of the folding free energy landscapes of metal-free versions of five ALS-inducing variants, A4V, L38V, G93A, L106V and S134N SOD1, were determined with a global analysis of kinetic and thermodynamic folding data for dimeric and stable monomeric versions of these variants. Utilizing this global analysis approach, the perturbations on the global stability in response to mutation can be partitioned between the monomer folding and association steps, and the effects of mutation on the populations of the folded and unfolded monomeric states can be determined. The 2- to 10-fold increase in the population of the folded monomeric state for A4V, L38V and L106V and the 80- to 480-fold increase in the population of the unfolded monomeric states for all but S134N would dramatically increase their propensity for aggregation through high-order nucleation reactions. The wild-type-like populations of these states for the metal-binding region S134N variant suggest that even wild-type SOD1 may also be prone to aggregation in the absence of metals.

Two mechanisms for dissipation of excess light in monomeric and trimeric light-harvesting complexes

Energy Technology Data Exchange (ETDEWEB)

Dall' Osto, Luca [Univ. di Verona, Verona (Italy). Dipartimento di Biotecnologie; Cazzaniga, Stefano [Univ. di Verona, Verona (Italy). Dipartimento di Biotecnologie; Bressan, Mauro [Univ. di Verona, Verona (Italy). Dipartimento di Biotecnologie; Paleček, David [Lund Univ. (Sweden). Dept. of Chemical Physics; Židek, Karel [Lund Univ. (Sweden). Dept. of Chemical Physics; Niyogi, Krishna K. [Univ. of California, Berkeley, CA (United States). Howard Hughes Medical Inst., Dept. of Plant and Microbial Biology; Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Molecular Biophysics and Integrated Bioimaging Division; Fleming, Graham R. [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Molecular Biophysics and Integrated Bioimaging Division; Univ. of California, Berkeley, CA (United States). Dept. of Chemistry, Graduate Group in Applied Science and Technology; Zigmantas, Donatas [Lund Univ. (Sweden). Dept. of Chemical Physics; Bassi, Roberto [Univ. di Verona, Verona (Italy). Dipartimento di Biotecnologie; Consiglio Nazionale delle Ricerche (CNR), Firenze (Italy). Istituto per la Protezione delle Piante (IPP)

2017-04-10

Oxygenic photoautotrophs require mechanisms for rapidly matching the level of chlorophyll excited states from light harvesting with the rate of electron transport from water to carbon dioxide. These photoprotective reactions prevent formation of reactive excited states and photoinhibition. The fastest response to excess illumination is the so-called non-photochemical quenching which, in higher plants, requires the luminal pH sensor PsbS and other yet unidentified components of the photosystem II antenna. Both trimeric light-harvesting complex II (LHCII) and monomeric LHC proteins have been indicated as site(s) of the heat-dissipative reactions. Different mechanisms have been proposed: Energy transfer to a lutein quencher in trimers, formation of a zeaxanthin radical cation in monomers. Here, we report on the construction of a mutant lacking all monomeric LHC proteins but retaining LHCII trimers. Its non-photochemical quenching induction rate was substantially slower with respect to the wild type. A carotenoid radical cation signal was detected in the wild type, although it was lost in the mutant. Here, we conclude that non-photochemical quenching is catalysed by two independent mechanisms, with the fastest activated response catalysed within monomeric LHC proteins depending on both zeaxanthin and lutein and on the formation of a radical cation. Trimeric LHCII was responsible for the slowly activated quenching component whereas inclusion in supercomplexes was not required. Finally, this latter activity does not depend on lutein nor on charge transfer events, whereas zeaxanthin was essential.

MxiN Differentially Regulates Monomeric and Oligomeric Species of the Shigella Type Three Secretion System ATPase Spa47.

Science.gov (United States)

Case, Heather B; Dickenson, Nicholas E

2018-04-17

Shigella rely entirely on the action of a single type three secretion system (T3SS) to support cellular invasion of colonic epithelial cells and to circumvent host immune responses. The ATPase Spa47 resides at the base of the Shigella needle-like type three secretion apparatus (T3SA), supporting protein secretion through the apparatus and providing a likely means for native virulence regulation by Shigella and a much needed target for non-antibiotic therapeutics to treat Shigella infections. Here, we show that MxiN is a differential regulator of Spa47 and that its regulatory impact is determined by the oligomeric state of the Spa47 ATPase, with which it interacts. In vitro and in vivo characterization shows that interaction of MxiN with Spa47 requires the six N-terminal residues of Spa47 that are also necessary for stable Spa47 oligomer formation and activation. This interaction with MxiN negatively influences the activity of Spa47 oligomers while upregulating the ATPase activity of monomeric Spa47. Detailed kinetic analyses of monomeric and oligomeric Spa47 in the presence and absence of MxiN uncover additional mechanistic insights into the regulation of Spa47 by MxiN, suggesting that the MxiN/Spa47 species resulting from interaction with monomeric and oligomeric Spa47 are functionally distinct and that both could be involved in Shigella T3SS regulation. Uncovering regulation of Spa47 by MxiN addresses an important gap in the current understanding of how Shigella controls T3SA activity and provides the first description of differential T3SS ATPase regulation by a native T3SS protein.

Crystallization and preliminary crystallographic analysis of decameric and monomeric forms of C49S mutant thioredoxin-dependent AhpC from Helicobacter pylori

International Nuclear Information System (INIS)

Supangat; Seo, Kyung Hye; Furqoni, Ahmad; Kwon, Young-Chul; Cho, Myung-Je; Rhee, Kwang-Ho; Lee, Sang Yeol; Lee, Kon Ho

2008-01-01

Decameric and monomeric forms of recombinant C49S mutant AhpC from H. pylori have been crystallized. Diffraction data were collected to 2.8 and 2.25 Å, respectively. Cys49Ser mutant Helicobacter pylori alkyl hydroperoxide reductase (C49S HpAhpC) was purified under reducing conditions in monomeric and decameric forms. The monomeric form was crystallized by the hanging-drop vapour-diffusion method. The crystals diffracted to 2.25 Å resolution and belonged to space group C2, with unit-cell parameters a = 245.8, b = 140.7, c = 189.5 Å, β = 127°, and contained 20 molecules in the asymmetric unit. A crystal of the decameric form was obtained by the microbatch crystallization method and diffracted to 2.8 Å resolution. It belonged to space group C222, with unit-cell parameters a = 257.5, b = 417.5, c = 95.6 Å. The structure of the monomeric form of C49S HpAhpC has been solved by the molecular-replacement method

Neurodevelopmental Expression Profile of Dimeric and Monomeric Group 1 mGluRs: Relevance to Schizophrenia Pathogenesis and Treatment.

Science.gov (United States)

Lum, Jeremy S; Fernandez, Francesca; Matosin, Natalie; Andrews, Jessica L; Huang, Xu-Feng; Ooi, Lezanne; Newell, Kelly A

2016-10-10

Group 1 metabotropic glutamate receptors (mGluR1/mGluR5) play an integral role in neurodevelopment and are implicated in psychiatric disorders, such as schizophrenia. mGluR1 and mGluR5 are expressed as homodimers, which is important for their functionality and pharmacology. We examined the protein expression of dimeric and monomeric mGluR1α and mGluR5 in the prefrontal cortex (PFC) and hippocampus throughout development (juvenile/adolescence/adulthood) and in the perinatal phencyclidine (PCP) model of schizophrenia. Under control conditions, mGluR1α dimer expression increased between juvenile and adolescence (209-328%), while monomeric levels remained consistent. Dimeric mGluR5 was steadily expressed across all time points; monomeric mGluR5 was present in juveniles, dramatically declining at adolescence and adulthood (-97-99%). The mGluR regulators, Homer 1b/c and Norbin, significantly increased with age in the PFC and hippocampus. Perinatal PCP treatment significantly increased juvenile dimeric mGluR5 levels in the PFC and hippocampus (37-50%) but decreased hippocampal mGluR1α (-50-56%). Perinatal PCP treatment also reduced mGluR1α dimer levels in the PFC at adulthood (-31%). These results suggest that Group 1 mGluRs have distinct dimeric and monomeric neurodevelopmental patterns, which may impact their pharmacological profiles at specific ages. Perinatal PCP treatment disrupted the early expression of Group 1 mGluRs which may underlie neurodevelopmental alterations observed in this model.

Survey of DOE NDA practices for CH-Tru waste certification--illustrated with a greater than 10,000 drum NDA data base

International Nuclear Information System (INIS)

Schultz, F.J.; Caldwell, J.T.; Smith, J.R.

1988-01-01

We have compiled a greater than 10,000 CH-TRU waste drum data base from seven DOE sites which have utilized such multiple NDA measurements within the past few years. Most of these nondestructive assay (NDA) technique assay result comparisons have been performed on well-characterized, segregated waste categories such as cemented sludges, combustibles, metals, graphite residues, glasses, etc., with well-known plutonium isotopic compositions. Waste segregation and categorization practices vary from one DOE site to another. Perhaps the most systematic approach has been in use for several years at the Rocky Flats Plant (RFP), operated by Rockwell International, and located near Golden, Colorado. Most of the drum assays in our data base result from assays of RFP wastes, with comparisons available between the original RFP assays and PAN assays performed independently at the Idaho National Engineering Laboratory (INEL) Solid Waste Examination Pilot Plant (SWEPP) facility. Most of the RFP assays were performed with hyperpure germanium (HPGe)-based SGS assay units. However, at least one very important waste category, processed first-stage sludges, is assayed at RFP using a sludge batch-sampling procedure, prior to filling of the waste drums. 5 refs., 5 figs

Structure/function analysis of PARP-1 in oxidative and nitrosative stress-induced monomeric ADPR formation.

Directory of Open Access Journals (Sweden)

Ben Buelow

2009-07-01

Full Text Available Poly adenosine diphosphate-ribose polymerase-1 (PARP-1 is a multifunctional enzyme that is involved in two major cellular responses to oxidative and nitrosative (O/N stress: detection and response to DNA damage via formation of protein-bound poly adenosine diphosphate-ribose (PAR, and formation of the soluble 2(nd messenger monomeric adenosine diphosphate-ribose (mADPR. Previous studies have delineated specific roles for several of PARP-1's structural domains in the context of its involvement in a DNA damage response. However, little is known about the relationship between the mechanisms through which PARP-1 participates in DNA damage detection/response and those involved in the generation of monomeric ADPR. To better understand the relationship between these events, we undertook a structure/function analysis of PARP-1 via reconstitution of PARP-1 deficient DT40 cells with PARP-1 variants deficient in catalysis, DNA binding, auto-PARylation, and PARP-1's BRCT protein interaction domain. Analysis of responses of the respective reconstituted cells to a model O/N stressor indicated that PARP-1 catalytic activity, DNA binding, and auto-PARylation are required for PARP-dependent mADPR formation, but that BRCT-mediated interactions are dispensable. As the BRCT domain is required for PARP-dependent recruitment of XRCC1 to sites of DNA damage, these results suggest that DNA repair and monomeric ADPR 2(nd messenger generation are parallel mechanisms through which PARP-1 modulates cellular responses to O/N stress.

A native promoter and inclusion of an intron is necessary for efficient expression of GFP or mRFP in Armillaria mellea

Science.gov (United States)

Armillaria mellea is a significant pathogen that causes Armillaria root disease on numerous hosts in forests, gardens and agricultural environments worldwide. Using a yeast-adapted pCAMBIA0380 Agrobacterium vector, we have constructed a series of vectors for transformation of A. mellea, assembled u...

MONOMERIC ß-AMYLOID INTERACTS WITH TYPE-1 INSULIN-LIKE GROWTH FACTOR RECEPTORS TO PROVIDE ENERGY SUPPLY TO NEURONS

Directory of Open Access Journals (Sweden)

Maria Laura eGiuffrida

2015-08-01

Full Text Available ß-amyloid (Aß1-42 is produced by proteolytic cleavage of the transmembrane type-1 protein, amyloid precursor protein. Under pathological conditions, Aß1-42 self-aggregates into oligomers, which cause synaptic dysfunction and neuronal loss, and are considered the culprit of Alzheimer’s disease (AD. However, Aß1-42 is mainly monomeric at physiological concentrations, and the precise role of monomeric Aß1-42 in neuronal function is largely unknown. We report that the monomer of Aß1-42 activates type-1 insulin-like growth factor receptors and enhances glucose uptake in neurons and peripheral cells by promoting the translocation of the Glut3 glucose transporter from the cytosol to the plasma membrane. In neurons, activity-dependent glucose uptake was blunted after blocking endogenous Aß production, and re-established in the presence of cerebrospinal fluid Aß. APP-null neurons failed to enhance depolarization-stimulated glucose uptake unless exogenous monomeric Aß1-42 was added. These data suggest that Aß1-42 monomers were critical for maintaining neuronal glucose homeostasis. Accordingly, exogenous Aß1-42 monomers were able to rescue the low levels of glucose consumption observed in brain slices from AD mutant mice.

Functional Characterization of Monomeric GTPase Rab1 in the Secretory Pathway of Leishmania*

Science.gov (United States)

Bahl, Surbhi; Parashar, Smriti; Malhotra, Himanshu; Raje, Manoj; Mukhopadhyay, Amitabha

2015-01-01

Leishmania secretes a large number of its effectors to the extracellular milieu. However, regulation of the secretory pathway in Leishmania is not well characterized. Here, we report the cloning, expression, and characterization of the Rab1 homologue from Leishmania. We have found that LdRab1 localizes in Golgi in Leishmania. To understand the role of LdRab1 in the secretory pathway of Leishmania, we have generated transgenic parasites overexpressing GFP-LdRab1:WT, GFP-LdRab1:Q67L (a GTPase-deficient dominant positive mutant of Rab1), and GFP-LdRab1:S22N (a GDP-locked dominant negative mutant of Rab1). Surprisingly, our results have shown that overexpression of GFP-LdRab1:Q67L or GFP-LdRab1:S22N does not disrupt the trafficking and localization of hemoglobin receptor in Leishmania. To determine whether the Rab1-dependent secretory pathway is conserved in parasites, we have analyzed the role of LdRab1 in the secretion of secretory acid phosphatase and Ldgp63 in Leishmania. Our results have shown that overexpression of GFP-LdRab1:Q67L or GFP-LdRab1:S22N significantly inhibits the secretion of secretory acid phosphatase by Leishmania. We have also found that overexpression of GFP-LdRab1:Q67L or GFP-LdRab1:S22N retains RFP-Ldgp63 in Golgi and blocks the secretion of Ldgp63, whereas the trafficking of RFP-Ldgp63 in GFP-LdRab1:WT-expressing cells is unaltered in comparison with control cells. Taken together, our results have shown that the Rab1-regulated secretory pathway is well conserved, and hemoglobin receptor trafficking follows an Rab1-independent secretory pathway in Leishmania. PMID:26499792

Functional Characterization of Monomeric GTPase Rab1 in the Secretory Pathway of Leishmania.

Science.gov (United States)

Bahl, Surbhi; Parashar, Smriti; Malhotra, Himanshu; Raje, Manoj; Mukhopadhyay, Amitabha

2015-12-11

Leishmania secretes a large number of its effectors to the extracellular milieu. However, regulation of the secretory pathway in Leishmania is not well characterized. Here, we report the cloning, expression, and characterization of the Rab1 homologue from Leishmania. We have found that LdRab1 localizes in Golgi in Leishmania. To understand the role of LdRab1 in the secretory pathway of Leishmania, we have generated transgenic parasites overexpressing GFP-LdRab1:WT, GFP-LdRab1:Q67L (a GTPase-deficient dominant positive mutant of Rab1), and GFP-LdRab1:S22N (a GDP-locked dominant negative mutant of Rab1). Surprisingly, our results have shown that overexpression of GFP-LdRab1:Q67L or GFP-LdRab1:S22N does not disrupt the trafficking and localization of hemoglobin receptor in Leishmania. To determine whether the Rab1-dependent secretory pathway is conserved in parasites, we have analyzed the role of LdRab1 in the secretion of secretory acid phosphatase and Ldgp63 in Leishmania. Our results have shown that overexpression of GFP-LdRab1:Q67L or GFP-LdRab1:S22N significantly inhibits the secretion of secretory acid phosphatase by Leishmania. We have also found that overexpression of GFP-LdRab1:Q67L or GFP-LdRab1:S22N retains RFP-Ldgp63 in Golgi and blocks the secretion of Ldgp63, whereas the trafficking of RFP-Ldgp63 in GFP-LdRab1:WT-expressing cells is unaltered in comparison with control cells. Taken together, our results have shown that the Rab1-regulated secretory pathway is well conserved, and hemoglobin receptor trafficking follows an Rab1-independent secretory pathway in Leishmania. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

Particle and energy balance in the SOL generated by a limiter in a RFP plasma

International Nuclear Information System (INIS)

Antoni, V.; Bagatin, M.; Desideri, D.; Serianni, G.

1992-01-01

The plasma outer region of the RFP experiment ETA BETA II has been extensively investigated. In particular by an insertable graphite limiter, instrumented with thermocouples and Langmuir probes, the energy and particle fluxes in the shadow of the limiter have been measured at different insertions. The results are compared with those obtained by small-sized calorimeter/Langmuir probes inserted to investigate, with fine space resolution, the outer region without limiter. Almost 80% of the power to the limiter is estimated to be carried by fast suprathermal electrons flowing along the magnetic field line direction. From an energy balance equation, applied in the SOL locally generated, the connection length of the limiter has been derived. Thus a particle balance has been applied to determine the particle diffusion coefficient at the plasma edge. (orig.)

The use of fluorescent intrabodies to detect endogenous gankyrin in living cancer cells

International Nuclear Information System (INIS)

Rinaldi, Anne-Sophie; Freund, Guillaume; Desplancq, Dominique; Sibler, Annie-Paule; Baltzinger, Mireille; Rochel, Natacha; Mély, Yves; Didier, Pascal; Weiss, Etienne

2013-01-01

Expression of antibody fragments in mammalian cells (intrabodies) is used to probe the target protein or interfere with its biological function. We previously described the in vitro characterisation of a single-chain Fv (scFv) antibody fragment (F5) isolated from an intrabody library that binds to the oncoprotein gankyrin (GK) in solution. Here, we have isolated several other scFvs that interact with GK in the presence of F5 and tested whether they allow, when fused to fluorescent proteins, to detect by FRET endogenous GK in living cells. The binding of pairs of scFvs to GK was analysed by gel filtration and the ability of each scFv to mediate nuclear import/export of GK was determined. Binding between scFv-EGFP and RFP-labelled GK in living cells was detected by fluorescence lifetime imaging microscopy (FLIM). After co-transfection of two scFvs fused to EGFP and RFP, respectively, which form a tri-molecular complex with GK in vitro, FRET signal was measured. This system allowed us to observe that GK is monomeric and distributed throughout the cytoplasm and nucleus of several cancer cell lines. Our results show that pairs of fluorescently labelled intrabodies can be monitored by FLIM–FRET microscopy and that this technique allows the detection of lowly expressed endogenous proteins in single living cells. Highlights: ► Endogenous GK in living cells was targeted with pairs of fluorescently-tagged scFvs. ► Tri-molecular complexes containing two scFvs and one molecule GK were formed. ► GK was detected using fluorescence lifetime-based FRET imaging. ► GK is monomeric and homogeneously distributed in several cancer cell lines. ► This technique may have many applications in live-cell imaging of endogenous proteins

The use of fluorescent intrabodies to detect endogenous gankyrin in living cancer cells

Energy Technology Data Exchange (ETDEWEB)

Rinaldi, Anne-Sophie; Freund, Guillaume; Desplancq, Dominique; Sibler, Annie-Paule; Baltzinger, Mireille [Ecole Supérieure de Biotechnologie de Strasbourg, UMR 7242, CNRS/Université de Strasbourg, boulevard Sébastien Brant, 67412 Illkirch (France); Rochel, Natacha [Institut de Génétique et de Biologie Moléculaire et Cellulaire, UMR 7104, CNRS/INSERM/Université de Strasbourg, rue Laurent Fries, 67404 Illkirch (France); Mély, Yves; Didier, Pascal [Faculté de Pharmacie, UMR 7213, CNRS/Université de Strasbourg, route du Rhin, 67401 Illkirch (France); Weiss, Etienne, E-mail: eweiss@unistra.fr [Ecole Supérieure de Biotechnologie de Strasbourg, UMR 7242, CNRS/Université de Strasbourg, boulevard Sébastien Brant, 67412 Illkirch (France)

2013-04-01

Expression of antibody fragments in mammalian cells (intrabodies) is used to probe the target protein or interfere with its biological function. We previously described the in vitro characterisation of a single-chain Fv (scFv) antibody fragment (F5) isolated from an intrabody library that binds to the oncoprotein gankyrin (GK) in solution. Here, we have isolated several other scFvs that interact with GK in the presence of F5 and tested whether they allow, when fused to fluorescent proteins, to detect by FRET endogenous GK in living cells. The binding of pairs of scFvs to GK was analysed by gel filtration and the ability of each scFv to mediate nuclear import/export of GK was determined. Binding between scFv-EGFP and RFP-labelled GK in living cells was detected by fluorescence lifetime imaging microscopy (FLIM). After co-transfection of two scFvs fused to EGFP and RFP, respectively, which form a tri-molecular complex with GK in vitro, FRET signal was measured. This system allowed us to observe that GK is monomeric and distributed throughout the cytoplasm and nucleus of several cancer cell lines. Our results show that pairs of fluorescently labelled intrabodies can be monitored by FLIM–FRET microscopy and that this technique allows the detection of lowly expressed endogenous proteins in single living cells. Highlights: ► Endogenous GK in living cells was targeted with pairs of fluorescently-tagged scFvs. ► Tri-molecular complexes containing two scFvs and one molecule GK were formed. ► GK was detected using fluorescence lifetime-based FRET imaging. ► GK is monomeric and homogeneously distributed in several cancer cell lines. ► This technique may have many applications in live-cell imaging of endogenous proteins.

Initial reversed-field pinch experiments on ZT-40 and recent advances in RFP theory

International Nuclear Information System (INIS)

Baker, D.A.; Buchenauer, C.J.; Burkhardt, L.C.

1980-01-01

The ZT-40 reversed-field pinch (RFP) has been operated in several modes: (1) without reversed toroidal field, (2) with self reversal, and (3) with aided reversal. An analytic ohmic heating and ignition model both confirm and provide guidance for transport codes. Nondissipative formation schemes have been analyzed and ideal MHD stable evolution and burn scenarios have been found. Particle and fluid simulations have produced qualitative agreement with respect to the nonlinear behavior of m = 0 resistive g-modes. Helical ohmic reversed field states are produced by a 2-D dynamical simulation, and nonlinear analytic work describes the final state. A fast resistive MHD code for linear stability has clarified the relations between several kinds of resistive instabilities. Ballooning modes and g-modes in systems with arbitrary magnetic shear including resistivity and viscosity, have been studied in a unified treatment with growth rate vs wavenumber showing the existence of important cutoffs

Monomeric, Oligomeric and Polymeric Proteins in Huntington Disease and Other Diseases of Polyglutamine Expansion

Directory of Open Access Journals (Sweden)

Guylaine Hoffner

2014-03-01

Full Text Available Huntington disease and other diseases of polyglutamine expansion are each caused by a different protein bearing an excessively long polyglutamine sequence and are associated with neuronal death. Although these diseases affect largely different brain regions, they all share a number of characteristics, and, therefore, are likely to possess a common mechanism. In all of the diseases, the causative protein is proteolyzed, becomes abnormally folded and accumulates in oligomers and larger aggregates. The aggregated and possibly the monomeric expanded polyglutamine are likely to play a critical role in the pathogenesis and there is increasing evidence that the secondary structure of the protein influences its toxicity. We describe here, with special attention to huntingtin, the mechanisms of polyglutamine aggregation and the modulation of aggregation by the sequences flanking the polyglutamine. We give a comprehensive picture of the characteristics of monomeric and aggregated polyglutamine, including morphology, composition, seeding ability, secondary structure, and toxicity. The structural heterogeneity of aggregated polyglutamine may explain why polyglutamine-containing aggregates could paradoxically be either toxic or neuroprotective.

Monomeric banana lectin at acidic pH overrules conformational stability of its native dimeric form.

Directory of Open Access Journals (Sweden)

Javed M Khan

Full Text Available Banana lectin (BL is a homodimeric protein categorized among jacalin-related family of lectins. The effect of acidic pH was examined on conformational stability of BL by using circular dichroism, intrinsic fluorescence, 1-anilino-8-napthalene sulfonate (ANS binding, size exclusion chromatography (SEC and dynamic light scattering (DLS. During acid denaturation of BL, the monomerization of native dimeric protein was found at pH 2.0. The elution profile from SEC showed two different peaks (59.65 ml & 87.98 ml at pH 2.0 while single peak (61.45 ml at pH 7.4. The hydrodynamic radii (R h of native BL was 2.9 nm while at pH 2.0 two species were found with R h of 1.7 and 3.7 nm. Furthermore at, pH 2.0 the secondary structures of BL remained unaltered while tertiary structure was significantly disrupted with the exposure of hydrophobic clusters confirming the existence of molten globule like state. The unfolding of BL with different subunit status was further evaluated by urea and temperature mediated denaturation to check their stability. As inferred from high Cm and ΔG values, the monomeric form of BL offers more resistance towards chemical denaturation than the native dimeric form. Besides, dimeric BL exhibited a Tm of 77°C while no loss in secondary structures was observed in monomers even up to 95°C. To the best of our knowledge, this is the first report on monomeric subunit of lectins showing more stability against denaturants than its native dimeric state.

mKikGR, a monomeric photoswitchable fluorescent protein.

Directory of Open Access Journals (Sweden)

Satoshi Habuchi

Full Text Available The recent demonstration and utilization of fluorescent proteins whose fluorescence can be switched on and off has greatly expanded the toolkit of molecular and cell biology. These photoswitchable proteins have facilitated the characterization of specifically tagged molecular species in the cell and have enabled fluorescence imaging of intracellular structures with a resolution far below the classical diffraction limit of light. Applications are limited, however, by the fast photobleaching, slow photoswitching, and oligomerization typical for photoswitchable proteins currently available. Here, we report the molecular cloning and spectroscopic characterization of mKikGR, a monomeric version of the previously reported KikGR that displays high photostability and switching rates. Furthermore, we present single-molecule imaging experiments that demonstrate that individual mKikGR proteins can be localized with a precision of better than 10 nanometers, suggesting their suitability for super-resolution imaging.

Selective defunctionalization by TiO2 of monomeric phenolics from lignin pyrolysis into simple phenols.

Science.gov (United States)

Mante, Ofei D; Rodriguez, Jose A; Babu, Suresh P

2013-11-01

This study is focused on defunctionalizing monomeric phenolics from lignin into simple phenols for applications such as phenol/formaldehyde resins, epoxidized novolacs, adhesives and binders. Towards this goal, Titanium dioxide (TiO2) was used to selectively remove hydroxyl, methoxy, carbonyl and carboxyl functionalities from the monomeric phenolic compounds from lignin to produce mainly phenol, cresols and xylenols. The results showed that anatase TiO2 was more selective and active compared to rutile TiO2. Catechols were found to be the most reactive phenolics and 4-ethylguaiacol the least reactive with anatase TiO2. An overall conversion of about 87% of the phenolics was achieved at 550°C with a catalyst-to-feed ratio of 5 w/w. Over 97% conversion of phenolics is achievable at moderate temperatures (550°C or ≤ 600°C) and a moderate catalyst-to-feed ratio of 6.5:1. The reactivity of catechols on TiO2 suggests that titania is a promising catalyst in the removal of hydroxyl moiety. Published by Elsevier Ltd.

Radial transport effects on ECCD in the TCV and DIII-D tokamaks and on Ohmic discharges in the MST RFP

International Nuclear Information System (INIS)

Harvey, R.W.; Sauter, O.; Nikkola, P.; Prater, R.; O'Connell, R.; Forest, C.B.

2003-01-01

The comprehensive CQL3D Fokker-Planck/Quasilinear simulation code has been benchmarked against experiment over a wide range of electron cyclotron conditions in the DIII-D tokamak (C.C. Petty et al., 14. Topical Conf. on RF Power in Plasmas, 2002). The same code, in disagreement with experiment, gives 560 kA of ECCD for a well documented, completely ECCD-driven, 100 kA TCV shot [O. Sauter et al, PRL, 2000]. Recent work (R.W. Harvey et al, Phys. Rev. Lett., 2002) has resolved the differences as due to radial transport at a level closely consistent with ITER scaling. Transport does not substantially affect DIII-D ECCD, but at similar ECH power has an overwhelming effect on the much smaller TCV. The transport is consistent with electrostatic-type diffusion (D ρρ constant in velocity space) and not with a magnetic-type diffusion (D ρρ ∝ |v || |). Fokker-Planck simulation of Ohmic reversed field pinch (RFP) discharges in the MST device reveals transport velocity dependence stronger than |v || |) will give agreement with current and soft X-ray spectra in standard discharges, but in the higher confinement, current profile controlled PPCD discharges, transport is again electrostatic-like. This is consistent with the object of PPCD, which is to replace magnetic turbulence driven current with auxiliary CD to improve transport. The tokamak and high-confinement RFP results mutually reinforce the constant-in-velocity-space 'electrostatic-type turbulence' conclusion. The steady-state energy and toroidal current are governed by the same radial transport equation. (authors)

RADIAL TRANSPORT EFFECTS ON ECCD IN THE TCV AND DIII-D TOKAMAKS AND ON OHMIC DISCHARGES IN THE MST RFP

International Nuclear Information System (INIS)

HARVEY, R.W.; SAUTER, O.; PRATER, R.; NIKKOLA, P.; O'CONNELL, R.; FOREST, C.B.

2002-01-01

The comprehensive CQL3D Fokker-Planck/Quasilinear simulation code has been benchmarked against experiment over a wide range of electron cyclotron conditions in the DIII-D tokamak (C.C. Petty et al., 14th Topical Conf. on RF Power in Plasmas, 2002). The same code, in disagreement with experiment, gives 560 kA of ECCD for a well documented, completely ECCD-driven, 100 kA TCV shot [O. Sauter et al, PRL, 2000]. Recent work (R.W. Harvey et al, Phys. Rev. Lett., 2002) has resolved the differences as due to radial transport at a level closely consistent with ITER scaling. Transport does not substantially affect DIII-D ECCD, but at similar ECH power has an overwhelming effect on the much smaller TCV. The transport is consistent with electrostatic-type diffusion (D ρρ constant in velocity-space) and not with a magnetic-type diffusion (D ρρ ∝ |v(parallel)|). Fokker-Planck simulation of Ohmic reversed field pinch (RFP) discharges in the MST device reveals transport velocity dependence stronger than |v(parallel)| will give agreement with current and soft X-ray spectra in standard discharges, but in the higher confinement, current profile controlled PPCD discharges, transport is again electrostatic-like. This is consistent with the object of PPCD, which is to replace magnetic turbulence driven current with auxiliary CD to improve transport. The tokamak and high-confinement RFP results mutually reinforce the constant-in-velocity-space ''electrostatic-type turbulence'' conclusion. The steady-state energy and toroidal current are governed by the same radial transport equation

Preparation and properties of a monomeric Mn(IV)-oxo complex.

Science.gov (United States)

Parsell, Trenton H; Behan, Rachel K; Green, Michael T; Hendrich, Michael P; Borovik, A S

2006-07-12

Manganese-oxo complexes have long been investigated because of their proposed roles in biological and chemical catalysis. However, there are few examples of monomeric complexes with terminal oxo ligands, especially those with oxomanganese(IV) units. A oxomanganese(IV) complex has been prepared from [MnIIIH3buea(O)]2- ([H3buea]3-, tris[(N'-tert-butylureaylato)-N-ethylene]aminato), a monomeric MnIII-O complex in which the oxo ligand arises from cleavage of dioxygen. Treating [MnIIIH3buea(O)]2- with [Cp2Fe]BF4 in either DMF at -45 degrees C or DMSO at room temperature produces [MnIVH3buea(O)]-: lambdamax = 635 nm; nu(Mn-16O) = 737 cm-1; nu(Mn-18O) = 709 cm-1; g = 5.15, 2.44, 1.63, D = 3.0 cm-1, E/D = 0.26, aMn = 66 G (A = 190 MHz). These spectroscopic properties support the assignment of a mononuclear MnIV-oxo complex with an S = 3/2 ground state. Density functional theory supports this assignment and the Jahn-Teller distortion around the high-spin MnIV center that would alter the molecular structure of [MnIVH3buea(O)]- from trigonal symmetry (as indicated by the highly rhombic EPR signal). [MnIVH3buea(O)]- is relatively unstable in DMSO, converting to [MnIIIH3buea(OH)]- via a proposed X-H bond cleavage. [MnIVH3buea(O)]- reacts with 1,2-diphenylhydrazine to from azobenzene (95% yield) and [MnIIIH3buea(OH)]-. The MnIV-oxo does not react with triphenyl- or tricyclohexylphosphine. However, O-atom transfer is observed with methyldiphenylphosphine and dimethylphenylphosphine, producing the corresponding phosphine oxides. These results illustrate the diverse reactivity of the MnIV-oxo unit.

Polyimide resin composites via in situ polymerization of monomeric reactants

Science.gov (United States)

Cavano, P. J.

1974-01-01

Thermo-oxidatively stable polyimide/graphite-fiber composites were prepared using a unique in situ polymerization of monomeric reactants directly on reinforcing fibers. This was accomplished by using an aromatic diamine and two ester-acids in a methyl alcohol solvent, rather than a previously synthesized prepolymer varnish, as with other A-type polyimides. A die molding procedure was developed and a composite property characterization conducted with high modulus graphite fiber tow. Flexure, tensile, compressive, and shear tests were conducted at temperatures from 72 to 650 F on laminates before and after exposures at the given temperatures in an air environment for times up to 1000 hours. The composite material was determined to be oxidatively, thermally, and hydrolytically stable.

Structural Characterization of Monomeric/Dimeric State of p59fyn SH2 Domain.

Science.gov (United States)

Huculeci, Radu; Kieken, Fabien; Garcia-Pino, Abel; Buts, Lieven; van Nuland, Nico; Lenaerts, Tom

2017-01-01

Src homology 2 (SH2) domains are key modulators in various signaling pathways allowing the recognition of phosphotyrosine sites of different proteins. Despite the fact that SH2 domains acquire their biological functions in a monomeric state, a multitude of reports have shown their tendency to dimerize. Here, we provide a technical description on how to isolate and characterize by gel filtration, circular dichroism (CD), and nuclear magnetic resonance (NMR) each conformational state of p59 fyn SH2 domain.

Comparison of excretory urographic contrast effects of dimeric and monomeric non-ionic iodinated contrast media in dogs

International Nuclear Information System (INIS)

Kishimoto, M.; Yamada, K.; Watanabe, A.; Miyamoto, K.; Iwasaki, T.; Miyake, Y.

2007-01-01

In excretory urography, the osmolarity of contrast media has rarely been treated as important in veterinary medicine. In this study, the contrast effect of two contrast media (monomeric iohexol and dimeric iodixanol) in the renal cortex and aorta were compared using computed tomography (CT). Five beagle dogs were used and the study employed a cross-over method for each contrast media. The results showed that there was no difference between the media in the aorta, but iodixanol showed higher CT value and a longer contrast effect than iohexol in the renal cortex, in spite of having the same iodine dosage. It is believed that iodixanol, with its low osmolarity, is diluted less by osmotic diuresis than monomeric iohexol. It is important to consider the osmolarity of the contrast media when evaluating the contrast effect, and it is essential to use the same contrast media for each examination, or the renal excretory speed will be under/overestimated

α-SNAP prevents docking of the acrosome during sperm exocytosis because it sequesters monomeric syntaxin.

Directory of Open Access Journals (Sweden)

Facundo Rodríguez

Full Text Available α-SNAP has an essential role in membrane fusion that consists of bridging cis SNARE complexes to NSF. α-SNAP stimulates NSF, which releases itself, α-SNAP, and individual SNAREs that subsequently re-engage in the trans arrays indispensable for fusion. α-SNAP also binds monomeric syntaxin and NSF disengages the α-SNAP/syntaxin dimer. Here, we examine why recombinant α-SNAP blocks secretion in permeabilized human sperm despite the fact that the endogenous protein is essential for membrane fusion. The only mammalian organism with a genetically modified α-SNAP is the hyh mouse strain, which bears a M105I point mutation; males are subfertile due to defective sperm exocytosis. We report here that recombinant α-SNAP-M105I has greater affinity for the cytosolic portion of immunoprecipitated syntaxin than the wild type protein and in consequence NSF is less efficient in releasing the mutant. α-SNAP-M105I is a more potent sperm exocytosis blocker than the wild type and requires higher concentrations of NSF to rescue its effect. Unlike other fusion scenarios where SNAREs are subjected to an assembly/disassembly cycle, the fusion machinery in sperm is tuned so that SNAREs progress uni-directionally from a cis configuration in resting cells to monomeric and subsequently trans arrays in cells challenged with exocytosis inducers. By means of functional and indirect immunofluorescense assays, we show that recombinant α-SNAPs--wild type and M105I--inhibit exocytosis because they bind monomeric syntaxin and prevent this SNARE from assembling with its cognates in trans. Sequestration of free syntaxin impedes docking of the acrosome to the plasma membrane assessed by transmission electron microscopy. The N-terminal deletion mutant α-SNAP-(160-295, unable to bind syntaxin, affects neither docking nor secretion. The implications of this study are twofold: our findings explain the fertility defect of hyh mice and indicate that assembly of SNAREs in trans

Sequence-specific 1H-NMR assignments for the aromatic region of several biologically active, monomeric insulins including native human insulin.

Science.gov (United States)

Roy, M; Lee, R W; Kaarsholm, N C; Thøgersen, H; Brange, J; Dunn, M F

1990-06-12

The aromatic region of the 1H-FT-NMR spectrum of the biologically fully-potent, monomeric human insulin mutant, B9 Ser----Asp, B27 Thr----Glu has been investigated in D2O. At 1 to 5 mM concentrations, this mutant insulin is monomeric above pH 7.5. Coupling and amino acid classification of all aromatic signals is established via a combination of homonuclear one- and two-dimensional methods, including COSY, multiple quantum filters, selective spin decoupling and pH titrations. By comparisons with other insulin mutants and with chemically modified native insulins, all resonances in the aromatic region are given sequence-specific assignments without any reliance on the various crystal structures reported for insulin. These comparisons also give the sequence-specific assignments of most of the aromatic resonances of the mutant insulins B16 Tyr----Glu, B27 Thr----Glu and B25 Phe----Asp and the chemically modified species des-(B23-B30) insulin and monoiodo-Tyr A14 insulin. Chemical dispersion of the assigned resonances, ring current perturbations and comparisons at high pH have made possible the assignment of the aromatic resonances of human insulin, and these studies indicate that the major structural features of the human insulin monomer (including those critical to biological function) are also present in the monomeric mutant.

Formation of amyloid fibers by monomeric light chain variable domains.

Science.gov (United States)

Brumshtein, Boris; Esswein, Shannon R; Landau, Meytal; Ryan, Christopher M; Whitelegge, Julian P; Phillips, Martin L; Cascio, Duilio; Sawaya, Michael R; Eisenberg, David S

2014-10-03

Systemic light chain amyloidosis is a lethal disease characterized by excess immunoglobulin light chains and light chain fragments composed of variable domains, which aggregate into amyloid fibers. These fibers accumulate and damage organs. Some light chains induce formation of amyloid fibers, whereas others do not, making it unclear what distinguishes amyloid formers from non-formers. One mechanism by which sequence variation may reduce propensity to form amyloid fibers is by shifting the equilibrium toward an amyloid-resistant quaternary structure. Here we identify the monomeric form of the Mcg immunoglobulin light chain variable domain as the quaternary unit required for amyloid fiber assembly. Dimers of Mcg variable domains remain stable and soluble, yet become prone to assemble into amyloid fibers upon disassociation into monomers. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

The Beckman DxI 800 prolactin assay demonstrates superior specificity for monomeric prolactin.

LENUS (Irish Health Repository)

Byrne, Brendan

2010-02-01

Commercially available prolactin immunoassays detect macroprolactin to variable degrees. Best practice requires laboratories to assess the cross-reactivity of their prolactin assay with macroprolactin, and where appropriate, introduce a screen for the presence of macroprolactin. Our policy has been to reanalyse hyperprolactinaemic samples following polyethylene glycol (PEG) precipitation and to report the resultant value as the monomeric prolactin content of the sample. The goal of this study was to determine the need to continue PEG precipitation when prolactin measurements with the Wallac AutoDELFIA were replaced by the Beckman DxI 800.

Extracellular Monomeric and Aggregated Tau Efficiently Enter Human Neurons through Overlapping but Distinct Pathways

Directory of Open Access Journals (Sweden)

Lewis D. Evans

2018-03-01

Full Text Available Summary: In Alzheimer’s disease, neurofibrillary tangle pathology appears to spread along neuronal connections, proposed to be mediated by the release and uptake of abnormal, disease-specific forms of microtubule-binding protein tau MAPT. It is currently unclear whether transfer of tau between neurons is a toxic gain-of-function process in dementia or reflects a constitutive biological process. We report two entry mechanisms for monomeric tau to human neurons: a rapid dynamin-dependent phase typical of endocytosis and a second, slower actin-dependent phase of macropinocytosis. Aggregated tau entry is independent of actin polymerization and largely dynamin dependent, consistent with endocytosis and distinct from macropinocytosis, the major route for aggregated tau entry reported for non-neuronal cells. Anti-tau antibodies abrogate monomeric tau entry into neurons, but less efficiently in the case of aggregated tau, where internalized tau carries antibody with it into neurons. These data suggest that tau entry to human neurons is a physiological process and not a disease-specific phenomenon. : In contrast with predictions that transfer of the microtubule-associated protein tau between neurons is a toxic gain-of-function process in dementia, Evans et al. show that healthy human neurons efficiently take up both normal and aggregated tau, by distinct but overlapping uptake mechanisms. Keywords: Alzheimer’s disease, frontotemporal dementia, Tau, MAPT, iPSC, endocytosis, human neurons, intracellular transport

Development of an immunotherapeutic adenovirus targeting hormone-independent prostate cancer

Directory of Open Access Journals (Sweden)

Kim JS

2013-11-01

Full Text Available Jae Sik Kim,1 Sang Don Lee,2 Sang Jin Lee,3 Moon Kee Chung21Department of Urology, The Catholic University of Korea Incheon St Mary's Hospital, Incheon, 2Pusan National University Yangsan Hospital and Research Institute for Convergence of Biomedical Science and Technology, Yangsan, 3Genitourinary Cancer Branch, National Cancer Center, Goyang, KoreaBackground: To develop a targeting therapy for hormone-independent prostate cancer, we constructed and characterized conditionally replicating oncolytic adenovirus (Ad equipped with mRFP(monomeric red fluorescence protein/ttk (modified herpes simplex virus thymidine kinase This construct was then further modified to express both mRFP/ttk and a soluble form of cytokine FLT3L (fms-related tyrosine kinase 3 ligand simultaneously.Methods: To construct the recombinant oncolytic adenovirus, E1a and E4 genes, which are necessary for adenovirus replication, were controlled by the prostate-specific enhancer sequence (PSES targeting prostate cancer cells expressing prostate-specific antigen (PSA and prostate-specific membrane antigen (PSMA. Simultaneously, it expressed the mRFP/ttk fusion protein in order to be able to elicit the cytotoxic effect.Results: The Ad5/35PSES.mRFP/ttk chimeric recombinant adenovirus was generated successfully. When replication of Ad5/35PSES.mRFP/ttk was evaluated in prostate cancer cell lines under fluorescence microscopy, red fluorescence intensity increased more in LNCaP cells, suggesting that the mRFP/ttk fusion protein was folded functionally. In addition, the replication assay including wild-type adenovirus as a positive control showed that PSES-positive cells (LNCaP and CWR22rv permitted virus replication but not PSES-negative cells (DU145 and PC3. Next, we evaluated the killing activity of this recombinant adenovirus. The Ad5/35PSES.mRFP/ttk killed LNCaP and CWR22rv more effectively. Unlike PSES-positive cells, DU145 and PC3 were resistant to killing by this recombinant

Activation and waste disposal of the TITAN RFP [reversed-field-pinch] reactors

International Nuclear Information System (INIS)

Cheng, E.T.; Conn, R.W.

1988-01-01

The TITAN-I lithium self-cooled and TITAN-II aqueous lithium nitrate solution-cooled fusion reactors are based on the reversed-field-pinch (RFP) toroidal confinement concept and operate at high power density with an 18.1 MW/m 2 neutron wall loading. These designs were analyzed to study the activation and waste disposal aspects of such high-power density reactors. It was found that because of the use of V-3Ti-1Si (TITAN-I) and reduced activation ferritic steel (TITAN-II) as structural alloys for the first wall, blanket, reflector, and shield components, all the TITAN components except the divertor collector plates can be classified as shallow-land burial (10CFR61 Class C or better) nuclear waste for disposal, provided that the impurity elements, niobium and molybdenum, can be controlled below about 1 and 0.3 appm levels, respectively. The average annual disposal masses were estimated to be 150 and 96 tonnes, respectively, for the 1,000 MW TITAN-I and TITAN-II reactors. This corresponds to about 11% of the total mass in the fusion power core of both reactors. The divertor collector plates are fabricated with tungsten because of its low particle sputtering properties. These divertor collector plates in the TITAN-I reactor will be qualified as Class C waste after 18.1 MW-y/m 2 operation. The waste disposal rating of the divertor collector plates in the TITAN-II reactor, however, is estimated to be a factor of 4 higher than allowed for Class C disposal, because of the soft neutron spectrum in the beryllium environment. The annual disposal mass of this non-Class C waste is 0.35 tons, about 0.04% of the average annual discharge mass for the TITAN-II reactor. An additional 74 m 3 annual discharge of Class C waste containing 14 C may be needed for the TITAN-II reactor because of the use of nitrate salt in the aqueous coolant as the tritium breeder. 13 refs., 6 tabs

The structure of mAG, a monomeric mutant of the green fluorescent protein Azami-Green, reveals the structural basis of its stable green emission

International Nuclear Information System (INIS)

Ebisawa, Tatsuki; Yamamura, Akihiro; Kameda, Yasuhiro; Hayakawa, Kou; Nagata, Koji; Tanokura, Masaru

2010-01-01

The crystal structure of a monomeric mutant of Azami-Green (mAG) from G. fascicularis was determined at 2.2 Å resolution. Monomeric Azami-Green (mAG) from the stony coral Galaxea fascicularis is the first known monomeric green-emitting fluorescent protein that is not a variant of Aequorea victoria green fluorescent protein (avGFP). These two green fluorescent proteins are only 27% identical in their amino-acid sequences. mAG is more similar in its amino-acid sequence to four fluorescent proteins: Dendra2 (a green-to-red irreversibly photoconverting fluorescent protein), Dronpa (a bright-and-dark reversibly photoswitchable fluorescent protein), KikG (a tetrameric green-emitting fluorescent protein) and Kaede (another green-to-red irreversibly photoconverting fluorescent protein). To reveal the structural basis of stable green emission by mAG, the 2.2 Å crystal structure of mAG has been determined and compared with the crystal structures of avGFP, Dronpa, Dendra2, Kaede and KikG. The structural comparison revealed that the chromophore formed by Gln62-Tyr63-Gly64 (QYG) and the fixing of the conformation of the imidazole ring of His193 by hydrogen bonds and van der Waals contacts involving His193, Arg66 and Thr69 are likely to be required for the stable green emission of mAG. The crystal structure of mAG will contribute to the design and development of new monomeric fluorescent proteins with faster maturation, brighter fluorescence, improved photostability, new colours and other preferable properties as alternatives to avGFP and its variants

Suppression of m = 0 in a RFP by toroidal field coils

International Nuclear Information System (INIS)

Alexander, D.; Robertson, S.

1993-01-01

The Reversatron RFP is normally operated with the toroidal field coils connected in series. The time-integrated voltage applied to the circuit determines the sum of the fluxes linking each turn but not the flux within each turn. Each winding may have a different flux determined by the external drive and by currents within the plasma. A parallel connection of the field coils results in the flux within each coil being determined by the volt-seconds applied to the windings; thus the toroidal flux is the same within each coil. This configuration suppresses any toroidal variation in the toroidal flux and effectively reduces the level of the m = 0 component of the radial field. The m = 0 fluctuations are expected to arise due to nonlinear coupling of the m = 1 modes. A parallel connection of field coils is impractical due to the low impedance required for driving the coils. The authors have tested the effect of parallel connected coils by adding an auxiliary set of 36 coils. These are connected in parallel but are not connected to any supply. The toroidal flux is generated by the series-connected coils which generate voltage but not current in the parallel-connected coils. With the auxiliary coils, the discharge duration is increased from 500 to 550 μsec, the plasma current is increased from 50 kA to 60 kA, F is more negative, Θ is larger, and there is less shot-to-shot variation in the discharges. The m = 0 fluctuations measured by 43 surface coils are, however, only slightly reduced

Antioxidant effects of phenolic rye (Secale cereale L.) extracts, monomeric hydroxycinnamates, and ferulic acid dehydrodimers on human low-density lipoproteins

DEFF Research Database (Denmark)

Andreasen, M.F.; Landbo, Anne-Katrine Regel; Christensen, L.P.

2001-01-01

Dietary antioxidants that protect low-density lipoprotein (LDL) from oxidation may help to prevent atherosclerosis and coronary heart disease. The antioxidant activities of purified monomeric and dimeric hydroxycinnamates and of phenolic extracts from rye (whole grain, bran, and flour) were...

Comparison of monomeric and polymeric horseradish peroxidase as labels in competitive ELISA for small molecule detection

International Nuclear Information System (INIS)

Li, Dongyang; Ying, Yibin; Wu, Jian; Niessner, Reinhard; Knopp, Dietmar

2013-01-01

We have developed a simple and sensitive competitive enzyme-linked immunosorbent assay (ELISA) to determine aflatoxin B1 (as a model small analyte) and using streptavidin-polymeric horseradish peroxidase complex (SApolyHRP) as a label for signal amplification. The performance of the assay was evaluated by comparing it with the classical indirect competitive ELISA using HRP labeled anti-mouse IgG as the tracer antibody. The results indicate that the SApolyHRP-based competitive ELISA exhibits a typically 2.4-fold steeper slope of the linear working range of the calibration curve compared to the monomeric HRP based classical ELISA, i.e., the sensitivity was increased. The SApolyHRP conjugate causes a typically 19-fold stronger signal generation in comparison to the traditional HRP labeled anti-mouse IgG at the same concentration (25 ng mL −1 ). Moreover, the SApolyHRP-based assay has a much wider linear range and a 3.8-fold better signal-to-noise ratio. Considering its simplicity, sensitivity and ease of operation, this competitive ELISA is considered to be a promising tool for small molecule immuno detection. (author)

A de novo designed monomeric, compact three helix bundle protein on a carbohydrate template

DEFF Research Database (Denmark)

Malik, Leila; Nygård, Jesper; Christensen, Niels Johan

2015-01-01

De novo design and chemical synthesis of proteins and of other artificial structures, which mimic them, is a central strategy for understanding protein folding and for accessing proteins with novel functions. We have previously described carbohydrates as templates for the assembly of artificial...... the template could facilitate protein folding. Here we report the design and synthesis of 3-helix bundle carboproteins on deoxy-hexopyranosides. The carboproteins were analyzed by CD, AUC, SAXS, and NMR, which revealed the formation of the first compact, and folded monomeric carboprotein distinctly different...

Antioxidant effects of phenolic rye (Secale cereale L.) extracts, monomeric hydroxycinnamates, and ferulic acid dehydrodimers on human low-density lipoproteins

DEFF Research Database (Denmark)

Andreasen, Mette Findal; Landbo, A K; Christensen, L P

2001-01-01

Dietary antioxidants that protect low-density lipoprotein (LDL) from oxidation may help to prevent atherosclerosis and coronary heart disease. The antioxidant activities of purified monomeric and dimeric hydroxycinnamates and of phenolic extracts from rye (whole grain, bran, and flour) were...... investigated using an in vitro copper-catalyzed human LDL oxidation assay. The most abundant ferulic acid dehydrodimer (diFA) found in rye, 8-O-4-diFA, was a slightly better antioxidant than ferulic acid and p-coumaric acid. The antioxidant activity of the 8-5-diFA was comparable to that of ferulic acid......, but neither 5-5-diFA nor 8-5-benzofuran-diFA inhibited LDL oxidation when added at 10-40 microM. The antioxidant activity of the monomeric hydroxycinnamates decreased in the following order: caffeic acid > sinapic acid > ferulic acid > p-coumaric acid. The antioxidant activity of rye extracts...

Hda monomerization by ADP binding promotes replicase clamp-mediated DnaA-ATP hydrolysis.

Science.gov (United States)

Su'etsugu, Masayuki; Nakamura, Kenta; Keyamura, Kenji; Kudo, Yuka; Katayama, Tsutomu

2008-12-26

ATP-DnaA is the initiator of chromosomal replication in Escherichia coli, and the activity of DnaA is regulated by the regulatory inactivation of the DnaA (RIDA) system. In this system, the Hda protein promotes DnaA-ATP hydrolysis to produce inactive ADP-DnaA in a mechanism that is mediated by the DNA-loaded form of the replicase sliding clamp. In this study, we first revealed that hda translation uses an unusual initiation codon, CUG, located downstream of the annotated initiation codon. The CUG initiation codon could be used for restricting the Hda level, as this initiation codon has a low translation efficiency, and the cellular Hda level is only approximately 100 molecules per cell. Hda translated using the correct reading frame was purified and found to have a high RIDA activity in vitro. Moreover, we found that Hda has a high affinity for ADP but not for other nucleotides, including ATP. ADP-Hda was active in the RIDA system in vitro and stable in a monomeric state, whereas apo-Hda formed inactive homomultimers. Both ADP-Hda and apo-Hda could form complexes with the DNA-loaded clamp; however, only ADP-Hda-DNA-clamp complexes were highly functional in the following interaction with DnaA. Formation of ADP-Hda was also observed in vivo, and mutant analysis suggested that ADP binding is crucial for cellular Hda activity. Thus, we propose that ADP is a crucial Hda ligand that promotes the activated conformation of the protein. ADP-dependent monomerization might enable the arginine finger of the Hda AAA+ domain to be accessible to ATP bound to the DnaA AAA+ domain.

Energy and Pitch Distribution of Spontaneously-generated High-energy Bulk Ions in the RFP

Science.gov (United States)

Kim, Jungha; Anderson, Jay; Reusch, Joshua; Eilerman, Scott; Capecchi, William

2014-10-01

Magnetic reconnection events in the reversed field pinch (RFP) are known to heat bulk and impurity ions. Runaway due to a parallel electric field has recently been confirmed as an important acceleration mechanism for high energy test ions supplied by a neutral beam. This effect does not, however, explain the change in distribution of nearly Maxwellian bulk ions at a reconnection event. By operating MST near maximum current and low electron density, significant fusion neutron flux can be generated without neutral beam injection. The bulk ion distribution created in these plasmas is well-confined, non-Maxwellian, and can be measured by the Advanced Neutral Particle Analyzer (ANPA) placed at a radial or tangential porthole. Data show a high energy tail up to 25 keV with a relatively higher signal in the low energy channels (8-15 keV) at the radial port following a reconnection event. Analysis of the energy dependence of trapped orbits sampled by the ANPA at the radial view implies an abundance of lower energy particles in regions of higher neutral density. This mandates a careful deconvolution of the measured ANPA signal to compute the fast ion distribution. This work is supported by the US DOE and NSF.

Resolved single-molecule detection of individual species within a mixture of anti-biotin antibodies using an engineered monomeric nanopore.

Science.gov (United States)

Fahie, Monifa; Chisholm, Christina; Chen, Min

2015-02-24

Oligomeric protein nanopores with rigid structures have been engineered for the purpose of sensing a wide range of analytes including small molecules and biological species such as proteins and DNA. We chose a monomeric β-barrel porin, OmpG, as the platform from which to derive the nanopore sensor. OmpG is decorated with seven flexible loops that move dynamically to create a distinct gating pattern when ionic current passes through the pore. Biotin was chemically tethered to the most flexible one of these loops. The gating characteristic of the loop's movement in and out of the porin was substantially altered by analyte protein binding. The gating characteristics of the pore with bound targets were remarkably sensitive to molecular identity, even providing the ability to distinguish between homologues within an antibody mixture. A total of five gating parameters were analyzed for each analyte to create a unique fingerprint for each biotin-binding protein. Our exploitation of gating noise as a molecular identifier may allow more sophisticated sensor design, while OmpG's monomeric structure greatly simplifies nanopore production.

The feeding tube of cyst nematodes: characterisation of protein exclusion.

Directory of Open Access Journals (Sweden)

Sebastian Eves-van den Akker

Full Text Available Plant parasitic nematodes comprise several groups; the most economically damaging of these are the sedentary endoparasites. Sedentary endoparasitic nematodes are obligate biotrophs and modify host root tissue, using a suite of effector proteins, to create a feeding site that is their sole source of nutrition. They feed by withdrawing host cell assimilate from the feeding site though a structure known as the feeding tube. The function, composition and molecular characteristics of feeding tubes are poorly characterised. It is hypothesised that the feeding tube facilitates uptake of host cell assimilate by acting as a molecular sieve. Several studies, using molecular mass as the sole indicator of protein size, have given contradictory results about the exclusion limits of the cyst nematode feeding tube. In this study we propose a method to predict protein size, based on protein database coordinates in silico. We tested the validity of these predictions using travelling wave ion mobility spectrometry--mass spectrometry, where predictions and measured values were within approximately 6%. We used the predictions, coupled with mass spectrometry, analytical ultracentrifugation and protein electrophoresis, to resolve previous conflicts and define the exclusion characteristics of the cyst nematode feeding tube. Heterogeneity was tested in the liquid, solid and gas phase to provide a comprehensive evaluation of three proteins of particular interest to feeding tube size exclusion, GFP, mRFP and Dual PI. The data and procedures described here could be applied to the design of plant expressed defence compounds intended for uptake into cyst nematodes. We also highlight the need to assess protein heterogeneity when creating novel fusion proteins.

The feeding tube of cyst nematodes: characterisation of protein exclusion.

Science.gov (United States)

Eves-van den Akker, Sebastian; Lilley, Catherine J; Ault, James R; Ashcroft, Alison E; Jones, John T; Urwin, Peter E

2014-01-01

Plant parasitic nematodes comprise several groups; the most economically damaging of these are the sedentary endoparasites. Sedentary endoparasitic nematodes are obligate biotrophs and modify host root tissue, using a suite of effector proteins, to create a feeding site that is their sole source of nutrition. They feed by withdrawing host cell assimilate from the feeding site though a structure known as the feeding tube. The function, composition and molecular characteristics of feeding tubes are poorly characterised. It is hypothesised that the feeding tube facilitates uptake of host cell assimilate by acting as a molecular sieve. Several studies, using molecular mass as the sole indicator of protein size, have given contradictory results about the exclusion limits of the cyst nematode feeding tube. In this study we propose a method to predict protein size, based on protein database coordinates in silico. We tested the validity of these predictions using travelling wave ion mobility spectrometry--mass spectrometry, where predictions and measured values were within approximately 6%. We used the predictions, coupled with mass spectrometry, analytical ultracentrifugation and protein electrophoresis, to resolve previous conflicts and define the exclusion characteristics of the cyst nematode feeding tube. Heterogeneity was tested in the liquid, solid and gas phase to provide a comprehensive evaluation of three proteins of particular interest to feeding tube size exclusion, GFP, mRFP and Dual PI. The data and procedures described here could be applied to the design of plant expressed defence compounds intended for uptake into cyst nematodes. We also highlight the need to assess protein heterogeneity when creating novel fusion proteins.

Hda Monomerization by ADP Binding Promotes Replicase Clamp-mediated DnaA-ATP Hydrolysis*S⃞

Science.gov (United States)

Su'etsugu, Masayuki; Nakamura, Kenta; Keyamura, Kenji; Kudo, Yuka; Katayama, Tsutomu

2008-01-01

ATP-DnaA is the initiator of chromosomal replication in Escherichia coli, and the activity of DnaA is regulated by the regulatory inactivation of the DnaA (RIDA) system. In this system, the Hda protein promotes DnaA-ATP hydrolysis to produce inactive ADP-DnaA in a mechanism that is mediated by the DNA-loaded form of the replicase sliding clamp. In this study, we first revealed that hda translation uses an unusual initiation codon, CUG, located downstream of the annotated initiation codon. The CUG initiation codon could be used for restricting the Hda level, as this initiation codon has a low translation efficiency, and the cellular Hda level is only ∼100 molecules per cell. Hda translated using the correct reading frame was purified and found to have a high RIDA activity in vitro. Moreover, we found that Hda has a high affinity for ADP but not for other nucleotides, including ATP. ADP-Hda was active in the RIDA system in vitro and stable in a monomeric state, whereas apo-Hda formed inactive homomultimers. Both ADP-Hda and apo-Hda could form complexes with the DNA-loaded clamp; however, only ADP-Hda-DNA-clamp complexes were highly functional in the following interaction with DnaA. Formation of ADP-Hda was also observed in vivo, and mutant analysis suggested that ADP binding is crucial for cellular Hda activity. Thus, we propose that ADP is a crucial Hda ligand that promotes the activated conformation of the protein. ADP-dependent monomerization might enable the arginine finger of the Hda AAA+ domain to be accessible to ATP bound to the DnaA AAA+ domain. PMID:18977760

Comparative analysis of the heme iron electronic structure and stereochemistry in tetrameric rabbit hemoglobin and monomeric soybean leghemoglobin a using Mössbauer spectroscopy with a high velocity resolution

Science.gov (United States)

Alenkina, I. V.; Kumar, A.; Berkovsky, A. L.; Oshtrakh, M. I.

2018-02-01

A comparative study of tetrameric rabbit hemoglobin and monomeric soybean leghemoglobin a in the oxy- and deoxy-forms was carried out using 57Fe Mössbauer spectroscopy with a high velocity resolution in order to analyze the heme iron electronic structure and stereochemistry in relation to the Mössbauer hyperfine parameters. The Mössbauer spectra of tetrameric rabbit hemoglobin in both forms were fitted using two quadrupole doublets related to the 57Fe in ɑ- and β-subunits. In contrast, the Mössbauer spectra of monomeric soybean leghemoglobin a were fitted using: (i) two quadrupole doublets for the oxy-form related to two conformational states of the distal His E7 imidazole ring and different hydrogen bonding of oxygen molecule in the oxy-form and (ii) using three quadrupole doublets for deoxy-form related to three conformational states of the proximal His F8 imidazole ring. Small variations of Mössbauer hyperfine parameters related to small differences in the heme iron electronic structure and stereochemistry in tetrameric rabbit hemoglobin and monomeric soybean leghemoglobin a are discussed.

Fermentation of Arabinoxylan-Oligosaccharides, Oligofructose and their Monomeric Sugars by Hindgut Bacteria from Siberian Sturgeon and African Catfish in Batch Culture in vitro

NARCIS (Netherlands)

Geraylou, Z.; Rurangwa, E.; Wiele, van der T.; Courtin, C.M.; Delcour, J.A.; Buyse, J.; Ollevier, F.

2014-01-01

The in vitro fermentation of two Non-Digestible Oligosaccharide (NDO) preparations, Arabinoxylan- Oligosaccharides (AXOS) and Oligofructose (OF), and their respective monomeric sugars, xylose and fructose, were investigated by hindgut microbiota of two major aquaculture fish species, Siberian

Development of a novel cell-based assay system EPISSAY for screening epigenetic drugs and liposome formulated decitabine

International Nuclear Information System (INIS)

Lim, Sue Ping; Callen, David F; Kumar, Raman; Akkamsetty, Yamini; Wang, Wen; Ho, Kristen; Neilsen, Paul M; Walther, Diego J; Suetani, Rachel J; Prestidge, Clive

2013-01-01

Despite the potential of improving the delivery of epigenetic drugs, the subsequent assessment of changes in their epigenetic activity is largely dependent on the availability of a suitable and rapid screening bioassay. Here, we describe a cell-based assay system for screening gene reactivation. A cell-based assay system (EPISSAY) was designed based on a silenced triple-mutated bacterial nitroreductase TMnfsB fused with Red-Fluorescent Protein (RFP) expressed in the non-malignant human breast cell line MCF10A. EPISSAY was validated using the target gene TXNIP, which has previously been shown to respond to epigenetic drugs. The potency of a epigenetic drug model, decitabine, formulated with PEGylated liposomes was also validated using this assay system. Following treatment with DNA methyltransferase (DNMT) and histone deacetylase (HDAC) inhibitors such as decitabine and vorinostat, increases in RFP expression were observed, indicating expression of RFP-TMnfsB. The EPISSAY system was then used to test the potency of decitabine, before and after PEGylated liposomal encapsulation. We observed a 50% higher potency of decitabine when encapsulated in PEGylated liposomes, which is likely to be due to its protection from rapid degradation. The EPISSAY bioassay system provides a novel and rapid system to compare the efficiencies of existing and newly formulated drugs that reactivate gene expression

A Domain of Herpes Simplex Virus pUL33 Required To Release Monomeric Viral Genomes from Cleaved Concatemeric DNA.

Science.gov (United States)

Yang, Kui; Dang, Xiaoqun; Baines, Joel D

2017-10-15

Monomeric herpesvirus DNA is cleaved from concatemers and inserted into preformed capsids through the actions of the viral terminase. The terminase of herpes simplex virus (HSV) is composed of three subunits encoded by U L 15, U L 28, and U L 33. The U L 33-encoded protein (pU L 33) interacts with pU L 28, but its precise role in the DNA cleavage and packaging reaction is unclear. To investigate the function of pU L 33, we generated a panel of recombinant viruses with either deletions or substitutions in the most conserved regions of U L 33 using a bacterial artificial chromosome system. Deletion of 11 amino acids (residues 50 to 60 or residues 110 to 120) precluded viral replication, whereas the truncation of the last 10 amino acids from the pU L 33 C terminus did not affect viral replication or the interaction of pU L 33 with pU L 28. Mutations that replaced the lysine at codon 110 and the arginine at codon 111 with alanine codons failed to replicate, and the pU L 33 mutant interacted with pU L 28 less efficiently. Interestingly, genomic termini of the large (L) and small (S) components were detected readily in cells infected with these mutants, indicating that concatemeric DNA was cleaved efficiently. However, the release of monomeric genomes as assessed by pulsed-field gel electrophoresis was greatly diminished, and DNA-containing capsids were not observed. These results suggest that pU L 33 is necessary for one of the two viral DNA cleavage events required to release individual genomes from concatemeric viral DNA. IMPORTANCE This paper shows a role for pU L 33 in one of the two DNA cleavage events required to release monomeric genomes from concatemeric viral DNA. This is the first time that such a phenotype has been observed and is the first identification of a function of this protein relevant to DNA packaging other than its interaction with other terminase components. Copyright © 2017 Yang et al.

LRP1 Modulates APP Intraneuronal Transport and Processing in Its Monomeric and Dimeric State

Directory of Open Access Journals (Sweden)

Claus U. Pietrzik

2017-04-01

Full Text Available The low-density lipoprotein receptor-related protein 1, LRP1, interacts with APP and affects its processing. This is assumed to be mostly caused by the impact of LRP1 on APP endocytosis. More recently, also an interaction of APP and LRP1 early in the secretory pathway was reported whereat retention of LRP1 in the ER leads to decreased APP cell surface levels and in turn, to reduced Aβ secretion. Here, we extended the biochemical and immunocytochemical analyses by showing via live cell imaging analyses in primary neurons that LRP1 and APP are transported only partly in common (one third but to a higher degree in distinct fast axonal transport vesicles. Interestingly, co-expression of LRP1 and APP caused a change of APP transport velocities, indicating that LRP1 recruits APP to a specific type of fast axonal transport vesicles. In contrast lowered levels of LRP1 facilitated APP transport. We further show that monomeric and dimeric APP exhibit similar transport characteristics and that both are affected by LRP1 in a similar way, by slowing down APP anterograde transport and increasing its endocytosis rate. In line with this, a knockout of LRP1 in CHO cells and in primary neurons caused an increase of monomeric and dimeric APP surface localization and in turn accelerated shedding by meprin β and ADAM10. Notably, a choroid plexus specific LRP1 knockout caused a much higher secretion of sAPP dimers into the cerebrospinal fluid compared to sAPP monomers. Together, our data show that LRP1 functions as a sorting receptor for APP, regulating its cell surface localization and thereby its processing by ADAM10 and meprin β, with the latter exhibiting a preference for APP in its dimeric state.

Method for the isolation of biologically active monomeric immunoglobulin A from a plasma fraction.

Science.gov (United States)

Leibl, H; Tomasits, R; Wolf, H M; Eibl, M M; Mannhalter, J W

1996-04-12

A purification method for immunoglobulin A (IgA) yielding monomeric IgA with a purity of over 97% has been developed. This procedure uses ethanol-precipitated plasma (Cohn fraction III precipitate) as the starting material and includes heparin-Sepharose adsorption, dextran sulfate and ammonium sulfate precipitation, hydroxyapatite chromatography, batch adsorption by an anion-exchange matrix and gel permeation. Additional protein G Sepharose treatment leads to an IgA preparation of greater than 99% purity. The isolated IgA presented with an IgA subclass distribution, equivalent to IgA in unfractionated plasma, and was biologically active, as was shown by its ability to down-modulate Haemophilus influenzae-b-induced IL-6 secretion of human monocytes.

Action of Monomeric/Gemini Surfactants on Free Cells and Biofilm of Asaia lannensis

Directory of Open Access Journals (Sweden)

Anna Koziróg

2017-11-01

Full Text Available We investigated the biological activity of surfactants based on quaternary ammonium compounds: gemini surfactant hexamethylene-1,6-bis-(N,N-dimethyl-N-dodecylammonium bromide (C6, synthesized by the reaction of N,N-dimethyl-N-dodecylamine with 1,6-dibromohexane, and its monomeric analogue dodecyltrimethylammonium bromide (DTAB. The experiments were performed with bacteria Asaia lannensis, a common spoilage in the beverage industry. The minimal inhibitory concentration (MIC values were determined using the tube standard two-fold dilution method. The growth and adhesive properties of bacterial cells were studied in different culture media, and the cell viability was evaluated using plate count method. Both of the surfactants were effective against the bacterial strain, but the MIC of gemini compound was significantly lower. Both C6 and DTAB exhibited anti-adhesive abilities. Treatment with surfactants at or below MIC value decreased the number of bacterial cells that were able to form biofilm, however, the gemini surfactant was more effective. The used surfactants were also found to be able to eradicate mature biofilms. After 4 h of treatment with C6 surfactant at concentration 10 MIC, the number of bacterial cells was reduced by 91.8%. The results of this study suggest that the antibacterial activity of the gemini compound could make it an effective microbiocide against the spoilage bacteria Asaia sp. in both planktonic and biofilm stages.

Action of Monomeric/Gemini Surfactants on Free Cells and Biofilm of Asaia lannensis.

Science.gov (United States)

Koziróg, Anna; Kręgiel, Dorota; Brycki, Bogumił

2017-11-22

We investigated the biological activity of surfactants based on quaternary ammonium compounds: gemini surfactant hexamethylene-1,6-bis-( N,N -dimethyl- N -dodecylammonium bromide) (C6), synthesized by the reaction of N,N -dimethyl- N- dodecylamine with 1,6-dibromohexane, and its monomeric analogue dodecyltrimethylammonium bromide (DTAB). The experiments were performed with bacteria Asaia lannensis , a common spoilage in the beverage industry. The minimal inhibitory concentration (MIC) values were determined using the tube standard two-fold dilution method. The growth and adhesive properties of bacterial cells were studied in different culture media, and the cell viability was evaluated using plate count method. Both of the surfactants were effective against the bacterial strain, but the MIC of gemini compound was significantly lower. Both C6 and DTAB exhibited anti-adhesive abilities. Treatment with surfactants at or below MIC value decreased the number of bacterial cells that were able to form biofilm, however, the gemini surfactant was more effective. The used surfactants were also found to be able to eradicate mature biofilms. After 4 h of treatment with C6 surfactant at concentration 10 MIC, the number of bacterial cells was reduced by 91.8%. The results of this study suggest that the antibacterial activity of the gemini compound could make it an effective microbiocide against the spoilage bacteria Asaia sp. in both planktonic and biofilm stages.

Creating a monomeric endonuclease TALE-I-SceI with high specificity and low genotoxicity in human cells.

Science.gov (United States)

Lin, Jianfei; Chen, He; Luo, Ling; Lai, Yongrong; Xie, Wei; Kee, Kehkooi

2015-01-01

To correct a DNA mutation in the human genome for gene therapy, homology-directed repair (HDR) needs to be specific and have the lowest off-target effects to protect the human genome from deleterious mutations. Zinc finger nucleases, transcription activator-like effector nuclease (TALEN) and CRISPR-CAS9 systems have been engineered and used extensively to recognize and modify specific DNA sequences. Although TALEN and CRISPR/CAS9 could induce high levels of HDR in human cells, their genotoxicity was significantly higher. Here, we report the creation of a monomeric endonuclease that can recognize at least 33 bp by fusing the DNA-recognizing domain of TALEN (TALE) to a re-engineered homing endonuclease I-SceI. After sequentially re-engineering I-SceI to recognize 18 bp of the human β-globin sequence, the re-engineered I-SceI induced HDR in human cells. When the re-engineered I-SceI was fused to TALE (TALE-ISVB2), the chimeric endonuclease induced the same HDR rate at the human β-globin gene locus as that induced by TALEN, but significantly reduced genotoxicity. We further demonstrated that TALE-ISVB2 specifically targeted at the β-globin sequence in human hematopoietic stem cells. Therefore, this monomeric endonuclease has the potential to be used in therapeutic gene targeting in human cells. © The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research.

Membrane topology of Golgi-localized probable S-adenosylmethionine-dependent methyltransferase in tobacco (Nicotiana tabacum) BY-2 cells.

Science.gov (United States)

Liu, Jianping; Hayashi, Kyoko; Matsuoka, Ken

2015-01-01

S-adenosylmethionine (SAM)-dependent methyltransferases (MTases) transfer methyl groups to substrates. In this study, a novel putative tobacco SAM-MTase termed Golgi-localized methyl transferase 1 (GLMT1) has been characterized. GLMT1 is comprised of 611 amino acids with short N-terminal region, putative transmembrane region, and C-terminal SAM-MTase domain. Expression of monomeric red fluorescence protein (mRFP)-tagged protein in tobacco BY-2 cell indicated that GLMT1 is a Golgi-localized protein. Analysis of the membrane topology by protease digestion suggested that both C-terminal catalytic region and N-terminal region seem to be located to the cytosolic side of the Golgi apparatus. Therefore, GLMT1 might have a different function than the previously studied SAM-MTases in plants.

Monomeric Aβ(1-40) and Aβ(1-42) Peptides in Solution Adopt Very Similar Ramachandran Map Distributions That Closely Resemble Random Coil.

Science.gov (United States)

Roche, Julien; Shen, Yang; Lee, Jung Ho; Ying, Jinfa; Bax, Ad

2016-02-09

The pathogenesis of Alzheimer's disease is characterized by the aggregation and fibrillation of amyloid peptides Aβ(1-40) and Aβ(1-42) into amyloid plaques. Despite strong potential therapeutic interest, the structural pathways associated with the conversion of monomeric Aβ peptides into oligomeric species remain largely unknown. In particular, the higher aggregation propensity and associated toxicity of Aβ(1-42) compared to that of Aβ(1-40) are poorly understood. To explore in detail the structural propensity of the monomeric Aβ(1-40) and Aβ(1-42) peptides in solution, we recorded a large set of nuclear magnetic resonance (NMR) parameters, including chemical shifts, nuclear Overhauser effects (NOEs), and J couplings. Systematic comparisons show that at neutral pH the Aβ(1-40) and Aβ(1-42) peptides populate almost indistinguishable coil-like conformations. Nuclear Overhauser effect spectra collected at very high resolution remove assignment ambiguities and show no long-range NOE contacts. Six sets of backbone J couplings ((3)JHNHα, (3)JC'C', (3)JC'Hα, (1)JHαCα, (2)JNCα, and (1)JNCα) recorded for Aβ(1-40) were used as input for the recently developed MERA Ramachandran map analysis, yielding residue-specific backbone ϕ/ψ torsion angle distributions that closely resemble random coil distributions, the absence of a significantly elevated propensity for β-conformations in the C-terminal region of the peptide, and a small but distinct propensity for αL at K28. Our results suggest that the self-association of Aβ peptides into toxic oligomers is not driven by elevated propensities of the monomeric species to adopt β-strand-like conformations. Instead, the accelerated disappearance of Aβ NMR signals in D2O over H2O, particularly pronounced for Aβ(1-42), suggests that intermolecular interactions between the hydrophobic regions of the peptide dominate the aggregation process.

Small-angle X-ray scattering analysis reveals the ATP-bound monomeric state of the ATPase domain from the homodimeric MutL endonuclease, a GHKL phosphotransferase superfamily protein.

Science.gov (United States)

Iino, Hitoshi; Hikima, Takaaki; Nishida, Yuya; Yamamoto, Masaki; Kuramitsu, Seiki; Fukui, Kenji

2015-05-01

DNA mismatch repair is an excision system that removes mismatched bases chiefly generated by replication errors. In this system, MutL endonucleases direct the excision reaction to the error-containing strand of the duplex by specifically incising the newly synthesized strand. Both bacterial homodimeric and eukaryotic heterodimeric MutL proteins belong to the GHKL ATPase/kinase superfamily that comprises the N-terminal ATPase and C-terminal dimerization regions. Generally, the GHKL proteins show large ATPase cycle-dependent conformational changes, including dimerization-coupled ATP binding of the N-terminal domain. Interestingly, the ATPase domain of human PMS2, a subunit of the MutL heterodimer, binds ATP without dimerization. The monomeric ATP-bound state of the domain has been thought to be characteristic of heterodimeric GHKL proteins. In this study, we characterized the ATP-bound state of the ATPase domain from the Aquifex aeolicus MutL endonuclease, which is a homodimeric GHKL protein unlike the eukaryotic MutL. Gel filtration, dynamic light scattering, and small-angle X-ray scattering analyses clearly showed that the domain binds ATP in a monomeric form despite its homodimeric nature. This indicates that the uncoupling of dimerization and ATP binding is a common feature among bacterial and eukaryotic MutL endonucleases, which we suggest is closely related to the molecular mechanisms underlying mismatch repair.

Pleiotropic benefit of monomeric and oligomeric flavanols on vascular health--a randomized controlled clinical pilot study.

Science.gov (United States)

Weseler, Antje R; Ruijters, Erik J B; Drittij-Reijnders, Marie-José; Reesink, Koen D; Haenen, Guido R M M; Bast, Aalt

2011-01-01

Cardiovascular diseases are expanding to a major social-economic burden in the Western World and undermine man's deep desire for healthy ageing. Epidemiological studies suggest that flavanol-rich foods (e.g. grapes, wine, chocolate) sustain cardiovascular health. For an evidenced-based application, however, sound clinical data on their efficacy are strongly demanded. In a double-blind, randomized, placebo-controlled intervention study we supplemented 28 male smokers with 200 mg per day of monomeric and oligomeric flavanols (MOF) from grape seeds. At baseline, after 4 and 8 weeks we measured macro- and microvascular function and a cluster of systemic biomarkers for major pathological processes occurring in the vasculature: disturbances in lipid metabolism and cellular redox balance, and activation of inflammatory cells and platelets. In the MOF group serum total cholesterol and LDL decreased significantly (P ≤ 0.05) by 5% (n = 11) and 7% (n = 9), respectively in volunteers with elevated baseline levels. Additionally, after 8 weeks the ratio of glutathione to glutathione disulphide in erythrocytes rose from baseline by 22% (n = 15, Pbenefit of an 8 weeks supplementation with 200 mg/d MOF in humans. ClinicalTrials.gov NCT00742287.

Interaction of Classical Platinum Agents with the Monomeric and Dimeric Atox1 Proteins: A Molecular Dynamics Simulation Study

Directory of Open Access Journals (Sweden)

Xiaolei Wang

2013-12-01

Full Text Available We carried out molecular dynamics simulations and free energy calculations for a series of binary and ternary models of the cisplatin, transplatin and oxaliplatin agents binding to a monomeric Atox1 protein and a dimeric Atox1 protein to investigate their interaction mechanisms. All three platinum agents could respectively combine with the monomeric Atox1 protein and the dimeric Atox1 protein to form a stable binary and ternary complex due to the covalent interaction of the platinum center with the Atox1 protein. The results suggested that the extra interaction from the oxaliplatin ligand–Atox1 protein interface increases its affinity only for the OxaliPt + Atox1 model. The binding of the oxaliplatin agent to the Atox1 protein might cause larger deformation of the protein than those of the cisplatin and transplatin agents due to the larger size of the oxaliplatin ligand. However, the extra interactions to facilitate the stabilities of the ternary CisPt + 2Atox1 and OxaliPt + 2Atox1 models come from the α1 helices and α2-β4 loops of the Atox1 protein–Atox1 protein interface due to the cis conformation of the platinum agents. The combinations of two Atox1 proteins in an asymmetric way in the three ternary models were analyzed. These investigations might provide detailed information for understanding the interaction mechanism of the platinum agents binding to the Atox1 protein in the cytoplasm.

Dose-finding study of carbamylated monomeric allergoid tablets in grass-allergic rhinoconjunctivitis patients.

Science.gov (United States)

Mösges, Ralph; Rohdenburg, Christina; Eichel, Andrea; Zadoyan, Gregor; Kasche, Elena-Manja; Shah-Hosseini, Kija; Lehmacher, Walter; Schmalz, Petra; Compalati, Enrico

2017-11-01

To determine the optimal effective and safe dose of sublingual immunotherapy tablets containing carbamylated monomeric allergoids in patients with grass pollen-induced allergic rhinoconjunctivitis. In this prospective, randomized, double-blind, active-controlled, multicenter, Phase II study, four different daily doses were applied preseasonally for 12 weeks. Of 158 randomized adults, 155 subjects (safety population) received 300 units of allergy (UA)/day (n = 36), 600 UA/day (n = 43), 1000 UA/day (n = 39), or 2000 UA/day (n = 37). After treatment, 54.3, 47.6, 59.0 and 51.4% of patients, respectively, ceased to react to the highest allergen concentration in a conjunctival provocation test. Furthermore, the response threshold improved in 70.4, 62.9, 76.7 and 66.7% of patients, respectively. No serious adverse events occurred. This study found 1000 UA/day to be the optimal effective and safe dose.

Structural Analysis of Monomeric RNA-Dependent Polymerases: Evolutionary and Therapeutic Implications.

Directory of Open Access Journals (Sweden)

Rodrigo Jácome

Full Text Available The crystal structures of monomeric RNA-dependent RNA polymerases and reverse transcriptases of more than 20 different viruses are available in the Protein Data Bank. They all share the characteristic right-hand shape of DNA- and RNA polymerases formed by the fingers, palm and thumb subdomains, and, in many cases, "fingertips" that extend from the fingers towards the thumb subdomain, giving the viral enzyme a closed right-hand appearance. Six conserved structural motifs that contain key residues for the proper functioning of the enzyme have been identified in all these RNA-dependent polymerases. These enzymes share a two divalent metal-ion mechanism of polymerization in which two conserved aspartate residues coordinate the interactions with the metal ions to catalyze the nucleotidyl transfer reaction. The recent availability of crystal structures of polymerases of the Orthomyxoviridae and Bunyaviridae families allowed us to make pairwise comparisons of the tertiary structures of polymerases belonging to the four main RNA viral groups, which has led to a phylogenetic tree in which single-stranded negative RNA viral polymerases have been included for the first time. This has also allowed us to use a homology-based structural prediction approach to develop a general three-dimensional model of the Ebola virus RNA-dependent RNA polymerase. Our model includes several of the conserved structural motifs and residues described in other viral RNA-dependent RNA polymerases that define the catalytic and highly conserved palm subdomain, as well as portions of the fingers and thumb subdomains. The results presented here help to understand the current use and apparent success of antivirals, i.e. Brincidofovir, Lamivudine and Favipiravir, originally aimed at other types of polymerases, to counteract the Ebola virus infection.

Highly sensitive and quantitative FRET-FLIM imaging in single dendritic spines using improved non-radiative YFP.

Science.gov (United States)

Murakoshi, Hideji; Lee, Seok-Jin; Yasuda, Ryohei

2008-08-01

Two-photon fluorescence lifetime imaging microscopy (TPFLIM) enables the quantitative measurements of fluorescence resonance energy transfer (FRET) in small subcellular compartments in light scattering tissue. We evaluated and optimized the FRET pair of mEGFP (monomeric EGFP with the A206K mutation) and REACh (non-radiative YFP variants) for TPFLIM. We characterized several mutants of REACh in terms of their "darkness," and their ability to act as a FRET acceptor for mEGFP in HeLa cells and hippocampal neurons. Since the commonly used monomeric mutation A206K increases the brightness of REACh, we introduced a different monomeric mutation (F223R) which does not affect the brightness. Also, we found that the folding efficiency of original REACh, as measured by the fluorescence lifetime of a mEGFP-REACh tandem dimer, was low and variable from cell to cell. Introducing two folding mutations (F46L, Q69M) into REACh increased the folding efficiency by approximately 50%, and reduced the variability of FRET signal. Pairing mEGFP with the new REACh (super-REACh, or sREACh) improved the signal-to-noise ratio compared to the mEGFP-mRFP or mEGFP-original REACh pair by approximately 50%. Using this new pair, we demonstrated that the fraction of actin monomers in filamentous and globular forms in single dendritic spines can be quantitatively measured with high sensitivity. Thus, the mEGFP-sREACh pair is suited for quantitative FRET measurement by TPFLIM, and enables us to measure protein-protein interactions in individual dendritic spines in brain slices with high sensitivity.

Alpha B- and βA3-crystallins containing d-aspartic acids exist in a monomeric state.

Science.gov (United States)

Sakaue, Hiroaki; Takata, Takumi; Fujii, Norihiko; Sasaki, Hiroshi; Fujii, Noriko

2015-01-01

Crystallin stability and subunit-subunit interaction are essential for eye lens transparency. There are three types of crystallins in lens, designated as α-, β-, and γ-crystallins. Alpha-crystallin is a hetero-polymer of about 800kDa, consisting of 35-40 subunits of two different αA- and αB-subunits, each of 20kDa. The β/γ-crystallin superfamily comprises oligomeric β-crystallin (2-6 subunits) and monomeric γ-crystallin. Since lens proteins have very long half-lives, they undergo numerous post-translational modifications including racemization, isomerization, deamidation, oxidation, glycation, and truncation, which may decrease crystallin solubility and ultimately cause cataract formation. Racemization and isomerization of aspartyl (Asp) residues have been detected only in polymeric α- and oligomeric β-crystallin, while the situation in monomeric γ-crystallin has not been studied. Here, we investigated the racemization and isomerization of Asp in the γ-crystallin fraction of elderly donors. The results show that Asp residues of γS-, γD- and γC-crystallins were not racemized and isomerized. However, strikingly, we found that a portion of αB-crystallin and βA3-crystallin moved to the lower molecular weight fraction which is the same size of γ-crystallin. In those fractions, Asp-96 of αB-crystallin and Asp-37 of βA3-crystallin were highly inverted, which do not occur in the native lens higher molecular weight fraction. Our results indicate the possibility that the inversion of Asp residues may induce dissociation of αB- and βA3-crystallins from the polymeric and oligomeric states. This is the first report that stereoinversion of amino acids disturbs lens protein assembly in aged human lens. Copyright © 2014 Elsevier B.V. All rights reserved.

Crystallization and preliminary X-ray analysis of a monomeric mutant of Azami-Green (mAG), an Aequorea victoria green fluorescent protein-like green-emitting fluorescent protein from the stony coral Galaxea fascicularis

International Nuclear Information System (INIS)

Ebisawa, Tatsuki; Yamamura, Akihiro; Kameda, Yasuhiro; Hayakawa, Kou; Nagata, Koji; Tanokura, Masaru

2009-01-01

A monomeric mutant of Azami-Green from G. fascicularis was expressed, purified and crystallized using the sitting-drop vapour-diffusion method. The crystal belonged to space group P1 and diffracted X-rays to 2.20 Å resolution. Monomeric Azami-Green (mAG) from the stony coral Galaxea fascicularis is the first monomeric green-emitting fluorescent protein that is not a derivative of Aequorea victoria green fluorescent protein (avGFP). mAG and avGFP are 27% identical in amino-acid sequence. Diffraction-quality crystals of recombinant mAG were obtained by the sitting-drop vapour-diffusion method using PEG 3350 as the precipitant. The mAG crystal diffracted X-rays to 2.20 Å resolution on beamline AR-NW12A at the Photon Factory (Tsukuba, Japan). The crystal belonged to space group P1, with unit-cell parameters a = 41.78, b = 51.72, c = 52.89 Å, α = 90.96, β = 103.41, γ = 101.79°. The Matthews coefficient (V M = 2.10 Å 3 Da −1 ) indicated that the crystal contained two mAG molecules per asymmetric unit

Very bright orange fluorescent plants: endoplasmic reticulum targeting of orange fluorescent proteins as visual reporters in transgenic plants

Directory of Open Access Journals (Sweden)

Mann David GJ

2012-05-01

Full Text Available Abstract Background The expression of fluorescent protein (FP genes as real-time visual markers, both transiently and stably, has revolutionized plant biotechnology. A palette of colors of FPs is now available for use, but the diversity has generally been underutilized in plant biotechnology. Because of the green and far-red autofluorescent properties of many plant tissues and the FPs themselves, red and orange FPs (RFPs, and OFPs, respectfully appear to be the colors with maximum utility in plant biotechnology. Within the color palette OFPs have emerged as the brightest FP markers in the visible spectra. This study compares several native, near-native and modified OFPs for their “brightness” and fluorescence, therefore, their usability as marker genes in transgenic plant tissues. Results The OFPs DsRed2, tdTomato, mOrange and pporRFP were all expressed under the control of the CaMV 35S promoter in agroinfiltration-mediated transient assays in Nicotiana benthamiana. Each of these, as well as endoplasmic reticulum (ER-targeted versions, were stably expressed in transgenic Nicotiana tabacum and Arabidopsis thaliana. Congruent results were observed between transient and stable assays. Our results demonstrated that there are several adequate OFP genes available for plant transformation, including the new pporRFP, an unaltered tetramer from the hard coral Porites porites. When the tandem dimer tdTomato and the monomeric mOrange were targeted to the ER, dramatic, ca. 3-fold, increase in plant fluorescence was observed. Conclusions From our empirical data, and a search of the literature, it appears that tdTomato-ER and mOrange-ER are the two highest fluorescing FPs available as reporters for transgenic plants. The pporRFP is a brightly fluorescing tetramer, but all tetramer FPs are far less bright than the ER-targeted monomers we report here.

A comparative immunogenicity study in rabbits of disulfide-stabilized, proteolytically cleaved, soluble trimeric human immunodeficiency virus type 1 gp140, trimeric cleavage-defective gp140 and monomeric gp120

International Nuclear Information System (INIS)

Beddows, Simon; Franti, Michael; Dey, Antu K.; Kirschner, Marc; Iyer, Sai Prasad N.; Fisch, Danielle C.; Ketas, Thomas; Yuste, Eloisa; Desrosiers, Ronald C.; Klasse, Per Johan; Maddon, Paul J.; Olson, William C.; Moore, John P.

2007-01-01

The human immunodeficiency virus type 1 (HIV-1) surface envelope glycoprotein (Env) complex, a homotrimer containing gp120 surface glycoprotein and gp41 transmembrane glycoprotein subunits, mediates the binding and fusion of the virus with susceptible target cells. The Env complex is the target for neutralizing antibodies (NAbs) and is the basis for vaccines intended to induce NAbs. Early generation vaccines based on monomeric gp120 subunits did not confer protection from infection; one alternative approach is therefore to make and evaluate soluble forms of the trimeric Env complex. We have directly compared the immunogenicity in rabbits of two forms of soluble trimeric Env and monomeric gp120 based on the sequence of HIV-1 JR-FL . Both protein-only and DNA-prime, protein-boost immunization formats were evaluated, DNA-priming having little or no influence on the outcome. One form of trimeric Env was made by disrupting the gp120-gp41 cleavage site by mutagenesis (gp140 UNC ), the other contains an intramolecular disulfide bond to stabilize the cleaved gp120 and gp41 moieties (SOSIP.R6 gp140). Among the three immunogens, SOSIP.R6 gp140 most frequently elicited neutralizing antibodies against the homologous, neutralization-resistant strain, HIV-1 JR-FL . All three proteins induced NAbs against more sensitive strains, but the breadth of activity against heterologous primary isolates was limited. When antibodies able to neutralize HIV-1 JR-FL were detected, antigen depletion studies showed they were not directed at the V3 region but were targeted at other, undefined gp120 and also non-gp120 epitopes

Characterization of monomeric DNA-binding protein Histone H1 in Leishmania braziliensis.

Science.gov (United States)

Carmelo, Emma; González, Gloria; Cruz, Teresa; Osuna, Antonio; Hernández, Mariano; Valladares, Basilio

2011-08-01

Histone H1 in Leishmania presents relevant differences compared to higher eukaryote counterparts, such as the lack of a DNA-binding central globular domain. Despite that, it is apparently fully functional since its differential expression levels have been related to changes in chromatin condensation and infectivity, among other features. The localization and the aggregation state of L. braziliensis H1 has been determined by immunolocalization, mass spectrometry, cross-linking and electrophoretic mobility shift assays. Analysis of H1 sequences from the Leishmania Genome Database revealed that our protein is included in a very divergent group of histones H1 that is present only in L. braziliensis. An antibody raised against recombinant L. braziliensis H1 recognized specifically that protein by immunoblot in L. braziliensis extracts, but not in other Leishmania species, a consequence of the sequence divergences observed among Leishmania species. Mass spectrometry analysis and in vitro DNA-binding experiments have also proven that L. braziliensis H1 is monomeric in solution, but oligomerizes upon binding to DNA. Finally, despite the lack of a globular domain, L. braziliensis H1 is able to form complexes with DNA in vitro, with higher affinity for supercoiled compared to linear DNA.

A model of insulin fibrils derived from the x-ray crystal structure of a monomeric insulin (despentapeptide insulin).

Science.gov (United States)

Brange, J; Dodson, G G; Edwards, D J; Holden, P H; Whittingham, J L

1997-04-01

The crystal structure of despentapeptide insulin, a monomeric insulin, has been refined at 1.3 A spacing and subsequently used to predict and model the organization in the insulin fibril. The model makes use of the contacts in the densely packed despentapeptide insulin crystal, and takes into account other experimental evidence, including binding studies with Congo red. The dimensions of this model fibril correspond well with those measured experimentally, and the monomer-monomer contacts within the fibril are in accordance with the known physical chemistry of insulin fibrils. Using this model, it may be possible to predict mutations in insulin that might alleviate problems associated with fibril formation during insulin therapy.

Monomeric Aβ1–40 and Aβ1–42 Peptides in Solution Adopt Very Similar Ramachandran Map Distributions That Closely Resemble Random Coil

Science.gov (United States)

2016-01-01

The pathogenesis of Alzheimer’s disease is characterized by the aggregation and fibrillation of amyloid peptides Aβ1–40 and Aβ1–42 into amyloid plaques. Despite strong potential therapeutic interest, the structural pathways associated with the conversion of monomeric Aβ peptides into oligomeric species remain largely unknown. In particular, the higher aggregation propensity and associated toxicity of Aβ1–42 compared to that of Aβ1–40 are poorly understood. To explore in detail the structural propensity of the monomeric Aβ1–40 and Aβ1–42 peptides in solution, we recorded a large set of nuclear magnetic resonance (NMR) parameters, including chemical shifts, nuclear Overhauser effects (NOEs), and J couplings. Systematic comparisons show that at neutral pH the Aβ1–40 and Aβ1–42 peptides populate almost indistinguishable coil-like conformations. Nuclear Overhauser effect spectra collected at very high resolution remove assignment ambiguities and show no long-range NOE contacts. Six sets of backbone J couplings (3JHNHα, 3JC′C′, 3JC′Hα, 1JHαCα, 2JNCα, and 1JNCα) recorded for Aβ1–40 were used as input for the recently developed MERA Ramachandran map analysis, yielding residue-specific backbone ϕ/ψ torsion angle distributions that closely resemble random coil distributions, the absence of a significantly elevated propensity for β-conformations in the C-terminal region of the peptide, and a small but distinct propensity for αL at K28. Our results suggest that the self-association of Aβ peptides into toxic oligomers is not driven by elevated propensities of the monomeric species to adopt β-strand-like conformations. Instead, the accelerated disappearance of Aβ NMR signals in D2O over H2O, particularly pronounced for Aβ1–42, suggests that intermolecular interactions between the hydrophobic regions of the peptide dominate the aggregation process. PMID:26780756

Recombinant DNA derived monomeric insulin analogue: comparison with soluble human insulin in normal subjects.

Science.gov (United States)

Vora, J P; Owens, D R; Dolben, J; Atiea, J A; Dean, J D; Kang, S; Burch, A; Brange, J

1988-11-12

To compare the rate of absorption from subcutaneous tissue and the resulting hypoglycaemic effect of iodine-125 labelled soluble human insulin and a monomeric insulin analogue derived by recombinant DNA technology. Single blind randomised comparison of equimolar doses of 125I labelled soluble human insulin and insulin analogue. Study in normal people at a diabetes research unit and a university department of medical physics. Seven healthy male volunteers aged 20-39 not receiving any other drugs. After an overnight fast and a basal period of one hour two doses (0.05 and 0.1 U/kg) of 125I labelled soluble human insulin and insulin analogue were injected subcutaneously into the anterior abdominal wall on four separate days. To find a fast acting insulin for meal related requirements in insulin dependent diabetics. MEASUREMENTS and main results--Residual radioactivity at the injection site was measured continuously for the first two hours after injection of the 125I labelled preparations and thereafter for five minutes simultaneously with blood sampling. Frequent venous blood samples were obtained over six hours for determination of plasma immunoreactive insulin, insulin analogue, glucose, and glucagon values. Time to 50% of initial radioactivity at the injection site for the insulin analogue compared with soluble insulin was 61 v 135 minutes (p less than 0.05) with 0.05 U/kg and 67 v 145 minutes (p less than 0.001) with 0.1 U/kg. Concentrations in plasma increased faster after the insulin analogue compared with soluble insulin, resulting in higher plasma concentrations between 10 and 150 minutes (0.001 less than p less than 0.05) after 0.05 U/kg and between 40 and 360 minutes (0.001 less than p less than 0.05) after 0.1 U/kg. The hypoglycaemic response to insulin analogue was a plasma glucose nadir at 60 minutes with both doses compared with 90 and 120 minutes with soluble insulin at 0.5 and 0.1 U/kg respectively. The response of glucagon substantiated the earlier and

MODEL REQUEST FOR PROPOSALS TO PROVIDE ENERGY AND OTHER ATTRIBUTES FROM AN OFFSHORE WIND POWER PROJECT

Energy Technology Data Exchange (ETDEWEB)

Jeremy Firestone; Dawn Kurtz Crompton

2011-10-22

This document provides a model RFP for new generation. The 'base' RFP is for a single-source offshore wind RFP. Required modifications are noted should a state or utility seek multi-source bids (e.g., all renewables or all sources). The model is premised on proposals meeting threshold requirements (e.g., a MW range of generating capacity and a range in terms of years), RFP issuer preferences (e.g., likelihood of commercial operation by a date certain, price certainty, and reduction in congestion), and evaluation criteria, along with a series of plans (e.g., site, environmental effects, construction, community outreach, interconnection, etc.). The Model RFP places the most weight on project risk (45%), followed by project economics (35%), and environmental and social considerations (20%). However, if a multi-source RFP is put forward, the sponsor would need to either add per-MWh technology-specific, life-cycle climate (CO2), environmental and health impact costs to bid prices under the 'Project Economics' category or it should increase the weight given to the 'Environmental and Social Considerations' category.

Ratiometric fluorescence polarization as a cytometric functional parameter: theory and practice

International Nuclear Information System (INIS)

Yishai, Yitzhak; Fixler, Dror; Cohen-Kashi, Meir; Zurgil, Naomi; Deutsch, Mordechai

2003-01-01

The use of ratiometric fluorescence polarization (RFP) as a functional parameter in monitoring cellular activation is suggested, based on the physical phenomenon of fluorescence polarization dependency on emission wavelengths in multiple (at least binary) solutions. The theoretical basis of this dependency is thoroughly discussed and examined via simulation. For simulation, aimed to imitate a fluorophore-stained cell, real values of the fluorescence spectrum and polarization of different single fluorophore solutions were used. The simulation as well as the experimentally obtained values of RFP indicated the high sensitivity of this measure. Finally, the RFP parameter was utilized as a cytometric measure in three exemplary cellular bioassays. In the first, the apoptotic effect of oxLDL in a human Jurkat FDA-stained T cell line was monitored by RFP. In the second, the interaction between cell surface membrane receptors of human T lymphocyte cells was monitored by RFP measurements as a complementary means to the fluorescence resonance energy transfer (FRET) technique. In the third bioassay, cellular thiol level of FDA- and CMFDA-labelled Jurkat T cells was monitored via RFP

Functional alteration of a dimeric insecticidal lectin to a monomeric antifungal protein correlated to its oligomeric status.

Directory of Open Access Journals (Sweden)

Nilanjana Banerjee

Full Text Available BACKGROUND: Allium sativum leaf agglutinin (ASAL is a 25-kDa homodimeric, insecticidal, mannose binding lectin whose subunits are assembled by the C-terminal exchange process. An attempt was made to convert dimeric ASAL into a monomeric form to correlate the relevance of quaternary association of subunits and their functional specificity. Using SWISS-MODEL program a stable monomer was designed by altering five amino acid residues near the C-terminus of ASAL. METHODOLOGY/PRINCIPAL FINDINGS: By introduction of 5 site-specific mutations (-DNSNN-, a β turn was incorporated between the 11(th and 12(th β strands of subunits of ASAL, resulting in a stable monomeric mutant ASAL (mASAL. mASAL was cloned and subsequently purified from a pMAL-c2X system. CD spectroscopic analysis confirmed the conservation of secondary structure in mASAL. Mannose binding assay confirmed that molecular mannose binds efficiently to both mASAL and ASAL. In contrast to ASAL, the hemagglutination activity of purified mASAL against rabbit erythrocytes was lost. An artificial diet bioassay of Lipaphis erysimi with mASAL displayed an insignificant level of insecticidal activity compared to ASAL. Fascinatingly, mASAL exhibited strong antifungal activity against the pathogenic fungi Fusarium oxysporum, Rhizoctonia solani and Alternaria brassicicola in a disc diffusion assay. A propidium iodide uptake assay suggested that the inhibitory activity of mASAL might be associated with the alteration of the membrane permeability of the fungus. Furthermore, a ligand blot assay of the membrane subproteome of R. solani with mASAL detected a glycoprotein receptor having interaction with mASAL. CONCLUSIONS/SIGNIFICANCE: Conversion of ASAL into a stable monomer resulted in antifungal activity. From an evolutionary aspect, these data implied that variable quaternary organization of lectins might be the outcome of defense-related adaptations to diverse situations in plants. Incorporation of m

Allergen-Specific Immunotherapy with Monomeric Allergoid in a Mouse Model of Atopic Dermatitis

Science.gov (United States)

Babakhin, Alexander; Andreev, Sergey; Nikonova, Alexandra; Shilovsky, Igor; Buzuk, Andrey; Elisyutina, Olga; Fedenko, Elena; Khaitov, Musa

2015-01-01

Atopic dermatitis (AD) is a widespread and difficult to treat allergic skin disease and is a tough challenge for healthcare. In this study, we investigated whether allergen-specific immunotherapy (ASIT) with a monomeric allergoid obtained by succinylation of ovalbumin (sOVA) is effective in a mouse model of atopic dermatitis. An experimental model of AD was reproduced by epicutaneous sensitization with ovalbumin (OVA). ASIT was performed with subcutaneous (SC) administration of increasing doses of OVA or sOVA. The levels of anti-OVA antibodies, as well as cytokines, were detected by ELISA. Skin samples from patch areas were taken for histologic examination. ASIT with either OVA or sOVA resulted in a reduction of both the anti-OVA IgE level and the IgG1/IgG2a ratio. Moreover, ASIT with sOVA increased the IFN-γ level in supernatants after splenocyte stimulation with OVA. Histologic analysis of skin samples from the sites of allergen application showed that ASIT improved the histologic picture by decreasing allergic inflammation in comparison with untreated mice. These data suggest that ASIT with a succinylated allergen represents promising approach for the treatment of AD. PMID:26275152

Sulfatide promotes the folding of proinsulin, preserves insulin crystals, and mediates its monomerization.

Science.gov (United States)

Osterbye, T; Jørgensen, K H; Fredman, P; Tranum-Jensen, J; Kaas, A; Brange, J; Whittingham, J L; Buschard, K

2001-06-01

Sulfatide is a glycolipid that has been associated with insulin-dependent diabetes mellitus. It is present in the islets of Langerhans and follows the same intracellular route as insulin. However, the role of sulfatide in the beta cell has been unclear. Here we present evidence suggesting that sulfatide promotes the folding of reduced proinsulin, indicating that sulfatide possesses molecular chaperone activity. Sulfatide associates with insulin by binding to the insulin domain A8--A10 and most likely by interacting with the hydrophobic side chains of the dimer-forming part of the insulin B-chain. Sulfatide has a dual effect on insulin. It substantially reduces deterioration of insulin hexamer crystals at pH 5.5, conferring stability comparable to those in beta cell granules. Sulfatide also mediates the conversion of insulin hexamers to the biological active monomers at neutral pH, the pH at the beta-cell surface. Finally, we report that inhibition of sulfatide synthesis with chloroquine and fumonisine B1 leads to inhibition of insulin granule formation in vivo. Our observations suggest that sulfatide plays a key role in the folding of proinsulin, in the maintenance of insulin structure, and in the monomerization process.

Toroidal fusion reactor design based on the reversed-field pinch

International Nuclear Information System (INIS)

Hagenson, R.L.

1978-07-01

The toroidal reversed-field pinch (RFP) achieves gross equilibrium and stability with a combination of high shear and wall stabilization, rather than the imposition of tokamak-like q-constraints. Consequently, confinement is provided primarily by poloidal magnetic fields, poloidal betas as large as approximately 0.58 are obtainable, the high ohmic-heating (toroidal) current densities promise a sole means of heating a D-T plasma to ignition, and the plasma aspect ratio is not limited by stability/equilibrium constraints. A reactor-like plasma model has been developed in order to quantify and to assess the general features of a power system based upon RFP confinement. An ''operating point'' has been generated on the basis of this plasma model and a relatively detailed engineering energy balance. These results are used to generate a conceptual engineering model of the reversed-field pinch reactor (RFPR) which includes a general description of a 750 MWe power plant and the preliminary consideration of vacuum/fueling, first wall, blanket, magnet coils, iron core, and the energy storage/transfer system

Development, validation and evaluation of an analytical method for the determination of monomeric and oligomeric procyanidins in apple extracts.

Science.gov (United States)

Hollands, Wendy J; Voorspoels, Stefan; Jacobs, Griet; Aaby, Kjersti; Meisland, Ane; Garcia-Villalba, Rocio; Tomas-Barberan, Francisco; Piskula, Mariusz K; Mawson, Deborah; Vovk, Irena; Needs, Paul W; Kroon, Paul A

2017-04-28

There is a lack of data for individual oligomeric procyanidins in apples and apple extracts. Our aim was to develop, validate and evaluate an analytical method for the separation, identification and quantification of monomeric and oligomeric flavanols in apple extracts. To achieve this, we prepared two types of flavanol extracts from freeze-dried apples; one was an epicatechin-rich extract containing ∼30% (w/w) monomeric (-)-epicatechin which also contained oligomeric procyanidins (Extract A), the second was an oligomeric procyanidin-rich extract depleted of epicatechin (Extract B). The parameters considered for method optimisation were HPLC columns and conditions, sample heating, mass of extract and dilution volumes. The performance characteristics considered for method validation included standard linearity, method sensitivity, precision and trueness. Eight laboratories participated in the method evaluation. Chromatographic separation of the analytes was best achieved utilizing a Hilic column with a binary mobile phase consisting of acidic acetonitrile and acidic aqueous methanol. The final method showed linearity for epicatechin in the range 5-100μg/mL with a correlation co-efficient >0.999. Intra-day and inter-day precision of the analytes ranged from 2 to 6% and 2 to 13% respectively. Up to dp3, trueness of the method was >95% but decreased with increasing dp. Within laboratory precision showed RSD values <5 and 10% for monomers and oligomers, respectively. Between laboratory precision was 4 and 15% (Extract A) and 7 and 30% (Extract B) for monomers and oligomers, respectively. An analytical method for the separation, identification and quantification of procyanidins in an apple extract was developed, validated and assessed. The results of the inter-laboratory evaluation indicate that the method is reliable and reproducible. Copyright © 2017. Published by Elsevier B.V.

Hydrogen production from the monomeric sugars hydrolyzed from hemicellulose by Enterobacter aerogenes

Energy Technology Data Exchange (ETDEWEB)

Ren, Yunli; Wang, Jianji; Liu, Zhen; Ren, Yunlai; Li, Guozhi [School of Chemical Engineering and Pharmaceutics, Henan University of Science and Technology, Luoyang 471039, Henan (China)

2009-12-15

Relatively large percentages of xylose with glucose, arabinose, mannose, galactose and rhamnose constitute the hydrolysis products of hemicellulose. In this paper, hydrogen production performance of facultative anaerobe (Enterobacter aerogenes) has been investigated from these different monomeric sugars except glucose. It was shown that the stereoisomers of mannose and galactose were more effective for hydrogen production than those of xylose and arabinose. The substrate of 5 g/l xylose resulted in a relative high level of hydrogen yield (73.8 mmol/l), hydrogen production efficiency (2.2 mol/mol) and a maximum hydrogen production rate (249 ml/l/h). The hydrogen yield, hydrogen production efficiency and the maximum hydrogen production rate reached 104 mmol/l, 2.35 mol/mol and 290 ml/l/h, respectively, on a substrate of 10 g/l galactose. The hydrogen yields and the maximum hydrogen production rates increased with an increase of mannose concentrations and reached 119 mmol/l and 518 ml/l/h on the culture of 25 g/l mannose. However, rhamnose was a relative poor carbon resource for E. aerogenes to produce hydrogen, from which the hydrogen yield and hydrogen production efficiency were about one half of that from the mannose substrate. E. aerogenes was found to be a promising strain for hydrogen production from hydrolysis products of hemicellulose. (author)

Design Function and Structure of a Monomeric CLC Transporter

Energy Technology Data Exchange (ETDEWEB)

L Robertson; L Kolmakova-Partensky; C Miller

2011-12-31

Channels and transporters of the ClC family cause the transmembrane movement of inorganic anions in service of a variety of biological tasks, from the unusual - the generation of the kilowatt pulses with which electric fish stun their prey - to the quotidian - the acidification of endosomes, vacuoles and lysosomes. The homodimeric architecture of ClC proteins, initially inferred from single-molecule studies of an elasmobranch Cl{sup -} channel and later confirmed by crystal structures of bacterial Cl{sup -}/H{sup +} antiporters, is apparently universal. Moreover, the basic machinery that enables ion movement through these proteins - the aqueous pores for anion diffusion in the channels and the ion-coupling chambers that coordinate Cl{sup -} and H{sup +} antiport in the transporters - are contained wholly within each subunit of the homodimer. The near-normal function of a bacterial ClC transporter straitjacketed by covalent crosslinks across the dimer interface and the behaviour of a concatemeric human homologue argue that the transport cycle resides within each subunit and does not require rigid-body rearrangements between subunits. However, this evidence is only inferential, and because examples are known in which quaternary rearrangements of extramembrane ClC domains that contribute to dimerization modulate transport activity, we cannot declare as definitive a 'parallel-pathways picture in which the homodimer consists of two single-subunit transporters operating independently. A strong prediction of such a view is that it should in principle be possible to obtain a monomeric ClC. Here we exploit the known structure of a ClC Cl{sup -}/H{sup +} exchanger, ClC-ec1 from Escherichia coli, to design mutants that destabilize the dimer interface while preserving both the structure and the transport function of individual subunits. The results demonstrate that the ClC subunit alone is the basic functional unit for transport and that cross-subunit interaction is not

The Escherichia coli P and Type 1 Pilus Assembly Chaperones PapD and FimC Are Monomeric in Solution

Energy Technology Data Exchange (ETDEWEB)

Sarowar, Samema; Hu, Olivia J.; Werneburg, Glenn T.; Thanassi, David G.; Li, Huilin; Christie, P. J.

2016-06-27

The chaperone/usher pathway is used by Gram-negative bacteria to assemble adhesive surface structures known as pili or fimbriae. Uropathogenic strains ofEscherichia coliuse this pathway to assemble P and type 1 pili, which facilitate colonization of the kidney and bladder, respectively. Pilus assembly requires a periplasmic chaperone and outer membrane protein termed the usher. The chaperone allows folding of pilus subunits and escorts the subunits to the usher for polymerization into pili and secretion to the cell surface. Based on previous structures of mutant versions of the P pilus chaperone PapD, it was suggested that the chaperone dimerizes in the periplasm as a self-capping mechanism. Such dimerization is counterintuitive because the chaperone G1 strand, important for chaperone-subunit interaction, is buried at the dimer interface. Here, we show that the wild-type PapD chaperone also forms a dimer in the crystal lattice; however, the dimer interface is different from the previously solved structures. In contrast to the crystal structures, we found that both PapD and the type 1 pilus chaperone, FimC, are monomeric in solution. Our findings indicate that pilus chaperones do not sequester their G1 β-strand by forming a dimer. Instead, the chaperones may expose their G1 strand for facile interaction with pilus subunits. We also found that the type 1 pilus adhesin, FimH, is flexible in solution while in complex with its chaperone, whereas the P pilus adhesin, PapGII, is rigid. Our study clarifies a crucial step in pilus biogenesis and reveals pilus-specific differences that may relate to biological function.

IMPORTANCEPili are critical virulence factors for many bacterial pathogens. UropathogenicE. colirelies on P and type 1 pili assembled by the chaperone/usher pathway to

A monomeric G protein-coupled receptor isolated in a high-density lipoprotein particle efficiently activates its G protein

DEFF Research Database (Denmark)

Whorton, Matthew R; Bokoch, Michael P; Rasmussen, Søren Gøgsig Faarup

2007-01-01

G protein-coupled receptors (GPCRs) respond to a diverse array of ligands, mediating cellular responses to hormones and neurotransmitters, as well as the senses of smell and taste. The structures of the GPCR rhodopsin and several G proteins have been determined by x-ray crystallography, yet...... the organization of the signaling complex between GPCRs and G proteins is poorly understood. The observations that some GPCRs are obligate heterodimers, and that many GPCRs form both homo- and heterodimers, has led to speculation that GPCR dimers may be required for efficient activation of G proteins. However......, technical limitations have precluded a definitive analysis of G protein coupling to monomeric GPCRs in a biochemically defined and membrane-bound system. Here we demonstrate that a prototypical GPCR, the beta2-adrenergic receptor (beta2AR), can be incorporated into a reconstituted high-density lipoprotein...

Crystal Structure of a Monomeric Form of Severe Acute Respiratory Syndrome Coronavirus Endonuclease Nsp15 Suggests a Role for Hexamerization As An Allosteric Switch

Energy Technology Data Exchange (ETDEWEB)

Joseph, J.S.; Saikatendu, K.S.; Subramanian, V.; Neuman, B.W.; Buchmeier, M.J.; Stevens, R.C.; Kuhn, P.; /Scripps Res. Inst.

2007-07-09

Mature nonstructural protein-15 (nsp15) from the severe acute respiratory syndrome coronavirus (SARS-CoV) contains a novel uridylate-specific Mn{sup 2+}-dependent endoribonuclease (NendoU). Structure studies of the full-length form of the obligate hexameric enzyme from two CoVs, SARS-CoV and murine hepatitis virus, and its monomeric homologue, XendoU from Xenopus laevis, combined with mutagenesis studies have implicated several residues in enzymatic activity and the N-terminal domain as the major determinant of hexamerization. However, the tight link between hexamerization and enzyme activity in NendoUs has remained an enigma. Here, we report the structure of a trimmed, monomeric form of SARS-CoV nsp15 (residues 28 to 335) determined to a resolution of 2.9 Angstroms. The catalytic loop (residues 234 to 249) with its two reactive histidines (His 234 and His 249) is dramatically flipped by {approx}120 degrees into the active site cleft. Furthermore, the catalytic nucleophile Lys 289 points in a diametrically opposite direction, a consequence of an outward displacement of the supporting loop (residues 276 to 295). In the full-length hexameric forms, these two loops are packed against each other and are stabilized by intimate intersubunit interactions. Our results support the hypothesis that absence of an adjacent monomer due to deletion of the hexamerization domain is the most likely cause for disruption of the active site, offering a structural basis for why only the hexameric form of this enzyme is active.

A vented pump limiter for the reversed field pinch RFX

International Nuclear Information System (INIS)

Sonato, P.

1998-01-01

The reversed field pinch (RFP) plasma performance, as in the Tokamak, is strongly correlated with the edge neutral particle control. The drawbacks of the conventional magnetic divertors and throat limiters on the RFP plasma have slackened the application of an active particle control system in existing devices. An advanced solution, based on the idea of the 'vented pump limiter' experimented on Tore Supra, has been conceived for RFX. This type of pump limiter is very attractive for a RFP. In this paper, the design of a 'vented limiter' prototype for RFX is presented. Up to six modules of this limiter can be installed at the equatorial plane of RFX, allowing a particle exhaust efficiency comparable with a divertor or a throat limiter working in a Tokamak. Finally, the optimization of this concept for the next step RFP device is presented. (orig.)

Monomeric RC-LH1 core complexes retard LH2 assembly and intracytoplasmic membrane formation in PufX-minus mutants of Rhodobacter sphaeroides.

Science.gov (United States)

Adams, Peter G; Mothersole, David J; Ng, Irene W; Olsen, John D; Hunter, C Neil

2011-09-01

In the model photosynthetic bacterium Rhodobacter sphaeroides domains of light-harvesting 2 (LH2) complexes surround and interconnect dimeric reaction centre-light-harvesting 1-PufX (RC-LH1-PufX) 'core' complexes, forming extensive networks for energy transfer and trapping. These complexes are housed in spherical intracytoplasmic membranes (ICMs), which are assembled in a stepwise process where biosynthesis of core complexes tends to dominate the early stages of membrane invagination. The kinetics of LH2 assembly were measured in PufX mutants that assemble monomeric core complexes, as a consequence of either a twelve-residue N-terminal truncation of PufX (PufXΔ12) or the complete removal of PufX (PufX(-)). Lower rates of LH2 assembly and retarded maturation of membrane invagination were observed for the larger and less curved ICM from the PufX(-) mutant, consistent with the proposition that local membrane curvature, initiated by arrays of bent RC-LH1-PufX dimers, creates a favourable environment for stable assembly of LH2 complexes. Transmission electron microscopy and high-resolution atomic force microscopy were used to examine ICM morphology and membrane protein organisation in these mutants. Some partitioning of core and LH2 complexes was observed in PufX(-) membranes, resulting in locally ordered clusters of monomeric RC-LH1 complexes. The distribution of core and LH2 complexes in the three types of membrane examined is consistent with previous models of membrane curvature and domain formation (Frese et al., 2008), which demonstrated that a combination of crowding and asymmetries in sizes and shapes of membrane protein complexes drives membrane organisation. 2011 Elsevier B.V. All rights reserved.

Introducing a Hydrogen-Bond Donor into a Weakly Nucleophilic Brønsted Base: Alkali Metal Hexamethyldisilazides (MHMDS, M=Li, Na, K, Rb and Cs) with Ammonia.

Science.gov (United States)

Neufeld, Roman; Michel, Reent; Herbst-Irmer, Regine; Schöne, Ralf; Stalke, Dietmar

2016-08-22

Alkali metal 1,1,1,3,3,3-hexamethyldisilazide (MHMDSs) are one of the most utilised weakly nucleophilic Brønsted bases in synthetic chemistry and especially in natural product synthesis. Like lithium organics, they aggregate depending on the employed donor solvents. Thus, they show different reactivity and selectivity as a function of their aggregation and solvation state. To date, monomeric LiHMDS with monodentate donor bases was only characterised in solution. Since the first preparation of LiHMDS in 1959 by Wannagat and Niederprüm, all efforts to crystallise monomeric LiHMDS in the absence of chelating ligands failed. Herein, we present ammonia adducts of LiHMDS, NaHMDS, KHMDS, RbHMDS and CsHMDS with unprecedented aggregation motifs: 1) The hitherto missing monomeric key compound in the LiHMDS aggregation architectures. Monomeric crystal structures of trisolvated LiHMDS (1) and NaHMDS (2), showing unique intermolecular hydrogen bonds, 2) the unprecedented tetrasolvated KHMDS (3) and RbHMDS (4) dimers and 3) the disolvated CsHMDS (5) dimer with very close intermolecular Si-CH3 ⋅⋅⋅Cs s-block "agostic" interactions have been prepared and characterised by single-crystal X-ray structure analysis. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Pushing the size limit of de novo structure ensemble prediction guided by sparse SDSL-EPR restraints to 200 residues: The monomeric and homodimeric forms of BAX

Science.gov (United States)

Fischer, Axel W.; Bordignon, Enrica; Bleicken, Stephanie; García-Sáez, Ana J.; Jeschke, Gunnar; Meiler, Jens

2016-01-01

Structure determination remains a challenge for many biologically important proteins. In particular, proteins that adopt multiple conformations often evade crystallization in all biologically relevant states. Although computational de novo protein folding approaches often sample biologically relevant conformations, the selection of the most accurate model for different functional states remains a formidable challenge, in particular, for proteins with more than about 150 residues. Electron paramagnetic resonance (EPR) spectroscopy can obtain limited structural information for proteins in well-defined biological states and thereby assist in selecting biologically relevant conformations. The present study demonstrates that de novo folding methods are able to accurately sample the folds of 192-residue long soluble monomeric Bcl-2-associated X protein (BAX). The tertiary structures of the monomeric and homodimeric forms of BAX were predicted using the primary structure as well as 25 and 11 EPR distance restraints, respectively. The predicted models were subsequently compared to respective NMR/X-ray structures of BAX. EPR restraints improve the protein-size normalized root-mean-square-deviation (RMSD100) of the most accurate models with respect to the NMR/crystal structure from 5.9 Å to 3.9 Å and from 5.7 Å to 3.3 Å, respectively. Additionally, the model discrimination is improved, which is demonstrated by an improvement of the enrichment from 5% to 15% and from 13% to 21%, respectively. PMID:27129417

Early cytokine modulation after the rapid induction phase of sublingual immunotherapy with mite monomeric allergoids.

Science.gov (United States)

Di Gioacchino, M; Perrone, A; Petrarca, C; Di Claudio, F; Mistrello, G; Falagiani, P; Dadorante, V; Verna, N; Braga, M; Ballone, E; Cavallucci, E

2008-01-01

The influence of different treatment schedules of sublingual immunotherapy (SLIT) in activating IL-10-producing T-cells, crucial in inducing allergen-specific tolerance, is not completely understood. The present work was designed to evaluate allergen driven interleukin release by mononuclear cells in the early phase of SLIT, after application of different induction schemes. Twenty mite-allergic patients were enrolled, 10 (group A) treated with a traditional 98 day induction scheme and 10 (group B) with a 16 day scheme with monomeric allergoid vaccine. At the end of the induction phase, the cumulative doses taken by group A and group B patients were equivalent to 50.5 and 50.3 microg of mite group 1 allergens, respectively. The release of Th1-, Th2- and Treg-related interleukins was assessed in culture supernatants of 5 microg/ml Der-p1-stimulated mononuclear cells, isolated before and after the induction phases. No relevant treatment-related side effects were observed. Interleukin release was similar in the two groups at the enrolment. Non-stimulated and Der p 1 stimulated release of studied cytokines was similar in the two groups at enrolment. Der p 1 stimulation significantly increased IL-10 release (pallergoids are utilized.

Reversed-Field Pinch plasma model

International Nuclear Information System (INIS)

Miley, G.H.; Nebel, R.A.; Moses, R.W.

1979-01-01

The stability of a Reversed-Field Pinch (RFP) is strongly dependent on the plasma profile and the confining sheared magnetic field. Magnetic diffusion and thermal transport produce changing conditions of stability. Despite the limited understanding of RFP transport, modelling is important to predict general trends and to study possible field programming options. To study the ZT-40 experiment and to predict the performance of future RFP reactors, a one-dimensional transport code has been developed. This code includes a linear, ideal MHD stability check based on an energy principle. The transport section integrates plasma profiles forward in time while the stability section periodically checks the stability of the evolving plasma profile

Women and family poultry production in rural Africa.

Science.gov (United States)

Gueye, E H

2000-02-01

Poultry production has existed for many generations in Africa, and almost every village household keeps chickens. The rural family poultry (RFP) are generally raised in free-range and/or backyard systems, which are traditional extensive husbandry systems. The development of an intensive poultry production has been the goal of the African government over the years. Despite efforts aiming for such goal, RFP is still very important in African countries that are both poor and net importers of food. It is a valuable asset because it can contribute significantly in alleviating poverty, securing food supply, and promoting gender equality. In view of this, interventions to improve RFP production systems should take into account the sociocultural issues, specifically gender-based aspects. It is noted that such interventions might, in addition to food security and poverty alleviation, also serve to promote gender equality. RFP development programs should be more women-friendly in order to facilitate women's participation, as RFP production in the region is generally a woman's business. Moreover, efforts to empower village women has to be envisaged cautiously as there is a serious risk of men taking over once the poultry sector becomes more profitable.

Enhanced rifampicin delivery to alveolar macrophages by solid lipid nanoparticles

Energy Technology Data Exchange (ETDEWEB)

Chuan Junlan [West China School of Pharmacy, Sichuan University, Key Laboratory of Drug Targeting and Drug Delivery System, Ministry of Education (China); Li Yanzhen [Tianjin Institute of Pharmaceutical Research, State Key Laboratory of Drug Delivery Technology and Pharmacokinetics (China); Yang Likai; Sun Xun [West China School of Pharmacy, Sichuan University, Key Laboratory of Drug Targeting and Drug Delivery System, Ministry of Education (China); Zhang Qiang [Peking University, State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences (China); Gong Tao, E-mail: gongtaoy@126.com; Zhang Zhirong, E-mail: zrzzl@vip.sina.com [West China School of Pharmacy, Sichuan University, Key Laboratory of Drug Targeting and Drug Delivery System, Ministry of Education (China)

2013-05-15

The present study aimed at developing a drug delivery system targeting the densest site of tuberculosis infection, the alveolar macrophages (AMs). Rifampicin (RFP)-loaded solid lipid nanoparticles (RFP-SLNs) with an average size of 829.6 {+-} 16.1 nm were prepared by a modified lipid film hydration method. The cytotoxicity of RFP-SLNs to AMs and alveolar epithelial type II cells (AECs) was examined using MTT assays. The viability of AMs and AECs was above 80 % after treatment with RFP-SLNs, which showed low toxicity to both AMs and AECs. Confocal Laser Scanning Microscopy was employed to observe the interaction between RFP-SLNs and both AMs and AECs. After incubating the cells with RFP-SLNs for 2 h, the fluorescent intensity in AMs was more and remained longer (from 0.5 to 12 h) when compared with that in AECs (from 0.5 to 8 h). In vitro uptake characteristics of RFP-SLNs in AMs and AECs were also investigated by detection of intracellular RFP by High performance liquid chromatography. Results showed that RFP-SLNs delivered markedly higher RFP into AMs (691.7 ng/mg in cultured AMs, 662.6 ng/mg in primary AMs) than that into AECs (319.2 ng/mg in cultured AECs, 287.2 ng/mg in primary AECs). Subsequently, in vivo delivery efficiency and the selectivity of RFP-SLNs were further verified in Sprague-Dawley rats. Under pulmonary administration of RFP-SLNs, the amount of RFP in AMs was significantly higher than that in AECs at each time point. Our results demonstrated that solid lipid nanoparticles are a promising strategy for the delivery of rifampicin to alveolar macrophages selectively.

Modulation of basophils' degranulation and allergy-related enzymes by monomeric and dimeric naphthoquinones.

Directory of Open Access Journals (Sweden)

Brígida R Pinho

Full Text Available Allergic disorders are characterized by an abnormal immune response towards non-infectious substances, being associated with life quality reduction and potential life-threatening reactions. The increasing prevalence of allergic disorders demands for new and effective anti-allergic treatments. Here we test the anti-allergic potential of monomeric (juglone, menadione, naphthazarin, plumbagin and dimeric (diospyrin and diosquinone naphthoquinones. Inhibition of RBL-2H3 rat basophils' degranulation by naphthoquinones was assessed using two complementary stimuli: IgE/antigen and calcium ionophore A23187. Additionally, we tested for the inhibition of leukotrienes production in IgE/antigen-stimulated cells, and studied hyaluronidase and lipoxidase inhibition by naphthoquinones in cell-free assays. Naphthazarin (0.1 µM decreased degranulation induced by IgE/antigen but not A23187, suggesting a mechanism upstream of the calcium increase, unlike diospyrin (10 µM that reduced degranulation in A23187-stimulated cells. Naphthoquinones were weak hyaluronidase inhibitors, but all inhibited soybean lipoxidase with the most lipophilic diospyrin, diosquinone and menadione being the most potent, thus suggesting a mechanism of competition with natural lipophilic substrates. Menadione was the only naphthoquinone reducing leukotriene C4 production, with a maximal effect at 5 µM. This work expands the current knowledge on the biological properties of naphthoquinones, highlighting naphthazarin, diospyrin and menadione as potential lead compounds for structural modification in the process of improving and developing novel anti-allergic drugs.

Gas-phase synthesis and structure of monomeric ZnOH: a model species for metalloenzymes and catalytic surfaces.

Science.gov (United States)

Zack, Lindsay N; Sun, Ming; Bucchino, Matthew P; Clouthier, Dennis J; Ziurys, Lucy M

2012-02-16

Monomeric ZnOH has been studied for the first time using millimeter and microwave gas-phase spectroscopy. ZnOH is important in surface processes and at the active site of the enzyme carbonic anhydrase. In the millimeter-wave direct-absorption experiments, ZnOH was synthesized by reacting zinc vapor, produced in a Broida-type oven, with water. In the Fourier-transform microwave measurements, ZnOH was produced in a supersonic jet expansion of CH(3)OH and zinc vapor, created by laser ablation. Multiple rotational transitions of six ZnOH isotopologues in their X(2)A' ground states were measured over the frequency range of 22-482 GHz, and splittings due to fine and hyperfine structure were resolved. An asymmetric top pattern was observed in the spectra, showing that ZnOH is bent, indicative of covalent bonding. From these data, spectroscopic constants and an accurate structure were determined. The Zn-O bond length was found to be similar to that in carbonic anhydrase and other model enzyme systems.

Resolution of two native monomeric 90 kDa nitrate reductase active proteins from Shewanella gelidimarina and the sequence of two napA genes

International Nuclear Information System (INIS)

Simpson, Philippa J.L.; McKinzie, Audra A.; Codd, Rachel

2010-01-01

Research highlights: → Two monomeric 90 kDa nitrate reductase active proteins from Shewanella gelidimarina. → Sequence of napA from napEDABC-type operon and napA from NapDAGHB-type operon. → Isolation of NAP as NapA or NapAB correlated with NapA P47E amino acid substitution. -- Abstract: The reduction of nitrate to nitrite in the bacterial periplasm occurs in the 90 kDa NapA subunit of the periplasmic nitrate reductase (NAP) system. Most Shewanella genomes contain two nap operons: napEDABC and napDAGHB, which is an unusual feature of this genus. Two native, monomeric, 90 kDa nitrate reductase active proteins were resolved by hydrophobic interaction chromatography from aerobic cultures of Shewanella gelidimarina replete with reduced nitrogen compounds. The 90 kDa protein obtained in higher yield was characterized as NapA by electronic absorption and electron paramagnetic resonance spectroscopies and was identified by LC/MS/MS and MALDI-TOF/TOF MS as NapA from the napEDABC-type operon. The other 90 kDa protein, which was unstable and produced in low yields, was posited as NapA from the napDAGHB-type operon. Two napA genes have been sequenced from the napEDABC-type and napDAGHB-type operons of S. gelidimarina. Native NAP from S. putrefaciens was resolved as one NapA monomer and one NapAB heterodimer. Two amino acid substitutions in NapA correlated with the isolation of NAP as a NapA monomer or a NapAB heterodimer. The resolution of native, redox-active NapA isoforms in Shewanella provides new insight into the respiratory versatility of this genus, which has implications in bioremediation and the assembly of microbial fuel cells.

[An experimental study on a slow-release complex with rifampicin-polylactic-co-glycolic acid-calcium 
phosphate cement].

Science.gov (United States)

Wu, Jianhuang; Ding, Zhou; Lei, Qing; Li, Miao; Liang, Yan; Lu, Tao

2016-09-28

To prepare the slow-release complex with rifampicin (RFP)-polylactic-co-glycolic acid (PLGA)-calcium phosphate cement (CPC) (RFP-PLGA-CPC complex), and to study its physical and chemical properties and drug release properties in vitro.
 The emulsification-solvent evaporation method was adopted to prepare rifampicin polylactic acid-glycolic acid (RFP-PLGA) slow-release microspheres, which were divided into 3 groups: a calcium phosphate bone cement group (CPC group), a CPC embedded with RFP group (RFP-CPC group), and a PLGA slow-release microspheres carrying RFP and the self-curing CPC group (RFP- PLGA-CPC complex group). The solidification time and porosity of materials were determined. The drug release experiments in vitro were carried out to observe the compressive strength, the change of section morphology before and after drug release. 
 The CPC group showed the shortest solidification time, while the RFP-PLGA-CPC complex group had the longest one. There was statistical difference in the porosity between the CPC group and the RFP-CPC group (Pbehavior of the complex, which was in accordance with zero order kinetics equation F=0.168×t.
 The porosity of RFP-PLGA-CPC complex is significantly higher than that of CPC, and it can keep slow release of the effective anti-tuberculosis drugs and maintain a certain mechanical strength for a long time.

Serum prolactin revisited: parametric reference intervals and cross platform evaluation of polyethylene glycol precipitation-based methods for discrimination between hyperprolactinemia and macroprolactinemia.

Science.gov (United States)

Overgaard, Martin; Pedersen, Susanne Møller

2017-10-26

Hyperprolactinemia diagnosis and treatment is often compromised by the presence of biologically inactive and clinically irrelevant higher-molecular-weight complexes of prolactin, macroprolactin. The objective of this study was to evaluate the performance of two macroprolactin screening regimes across commonly used automated immunoassay platforms. Parametric total and monomeric gender-specific reference intervals were determined for six immunoassay methods using female (n=96) and male sera (n=127) from healthy donors. The reference intervals were validated using 27 hyperprolactinemic and macroprolactinemic sera, whose presence of monomeric and macroforms of prolactin were determined using gel filtration chromatography (GFC). Normative data for six prolactin assays included the range of values (2.5th-97.5th percentiles). Validation sera (hyperprolactinemic and macroprolactinemic; n=27) showed higher discordant classification [mean=2.8; 95% confidence interval (CI) 1.2-4.4] for the monomer reference interval method compared to the post-polyethylene glycol (PEG) recovery cutoff method (mean=1.8; 95% CI 0.8-2.8). The two monomer/macroprolactin discrimination methods did not differ significantly (p=0.089). Among macroprolactinemic sera evaluated by both discrimination methods, the Cobas and Architect/Kryptor prolactin assays showed the lowest and the highest number of misclassifications, respectively. Current automated immunoassays for prolactin testing require macroprolactin screening methods based on PEG precipitation in order to discriminate truly from falsely elevated serum prolactin. While the recovery cutoff and monomeric reference interval macroprolactin screening methods demonstrate similar discriminative ability, the latter method also provides the clinician with an easy interpretable monomeric prolactin concentration along with a monomeric reference interval.

Sterically crowded monomeric neutral bis(benzamidinato) compounds of aluminium, [PhC(NSiMe(3))(2)](2)AlX (X=Cl, H); X-ray crystal structure of [PhC(NSiMe(3))(2)]2AlH

NARCIS (Netherlands)

Duchateau, R; Meetsma, A; Teuben, JH

1996-01-01

AlCl3 reacts with [PhC(NSiMe(3))(2)]Li(OEt(2)) to afford the bis(N,N'-bis(trimethylsilyl)benzamidinato)aluminium chloro compound which, on treatment with KBEt(3)H, yields the structurally characterized monomeric hydride derivative, [PhC(NSiMe(3))(2)]2AlH, whose reactivity towards unsaturated

Novel DOTA-based prochelator for divalent peptide vectorization: synthesis of dimeric bombesin analogues for multimodality tumor imaging and therapy.

Science.gov (United States)

Abiraj, Keelara; Jaccard, Hugues; Kretzschmar, Martin; Helm, Lothar; Maecke, Helmut R

2008-07-28

Dimeric peptidic vectors, obtained by the divalent grafting of bombesin analogues on a newly synthesized DOTA-based prochelator, showed improved qualities as tumor targeted imaging probes in comparison to their monomeric analogues.

Dynamics of red fluorescent dental plaque during experimental gingivitis--A cohort study.

Science.gov (United States)

van der Veen, Monique H; Volgenant, Catherine M C; Keijser, Bart; Ten Cate, Jacob Bob M; Crielaard, Wim

2016-05-01

The dynamics of red fluorescent plaque (RFP) in comparison to clinical plaque and bleeding scores were studied during an experimental gingivitis protocol in a cohort of healthy participants. Forty-one participants were monitored for RFP before (24h plaque), during 14 days plaque accumulation (days 2, 5, 9, 14) and after 7 days recovery (24h plaque). RFP was assessed on fluorescence photographs of the vestibular aspect of the anterior teeth (cuspid to cuspid) in the upper and lower jaw. Clinical plaque and bleeding were assessed at days -14, 0, 14 and 21. RFP of 24h plaque was reproducible (days -14, 0), then increased during 14 days plaque accumulation and returned to baseline after 7 days recovery. Groups of low, moderate and high RFP formers were statistically significantly different at all times even already at baseline. The individual RFP response during 14 days plaque accumulation correlated well with RFP of 24h plaque (days -14, 0). RFP correlated moderate to well with clinical plaque at days -14, 0, 14 and 21. From day 2 of the gingivitis challenge RFP correlated with bleeding at day 14. RFP provided an objective measure of oral hygiene status. Given the correlation with clinical parameters found, the amount of RFP after 24h plaque accumulation was indicatory for the inflammatory response during a prolonged period of no oral hygiene. This trial was registered at the public trial register ​of the Central Committee on Research Involving Human Subjects (CCMO) under number NL51111.029.14 CLINICAL SIGNIFICANCE: This paper shows the association between RFP after 24h plaque accumulation and inflammatory response after a prolonged period of no oral hygiene. Red plaque fluorescence can be used to identify subjects at risk for developing gingival inflammation. Copyright © 2016 Elsevier Ltd. All rights reserved.

MALDI-TOF mass spectrometric detection of multiplex single base extended primers

DEFF Research Database (Denmark)

Mengel-From, Jonas; Sanchez Sanchez, Juan Jose; Børsting, Claus

2004-01-01

of multiple SNPs in a single reaction. Biotin-labeled ddNTPs were used in the SBE reaction and solid phase-bound monomeric avidin was used as capturing/purification scheme allowing the exclusive release of the SBE products under gentle conditions using 5% triethylamine. We dubbed this method monomeric avidin...

Oxovanadium(IV) complexes with tridentate dibasic schiff base ligands and 2-(2'-pyridyl) benzimidazole

Energy Technology Data Exchange (ETDEWEB)

Mohanty, R N; Chakravortty, V; Dash, K C [Utkal Univ., Bhubaneswar (India). Dept. of Chemistry

1991-05-01

The present work deals with the monomeric, six-coordinated mixed-ligand complexes of oxovanadium(IV) with dibasic tridentate schiff base ligands(ONO donor set) and the bidentate chelating ligand 2-(2'-pyridyl)benzimidazole (PBH) containing N{sub 2} donor set. (author). 1 tab., 22 refs.

Structural design and manufacturing of TPE-RX vacuum vessel

Energy Technology Data Exchange (ETDEWEB)

Sago, H.; Orita, J.; Kaguchi, H.; Ishigami, Y. [Mitsubishi Heavy Ind. Ltd., Kobe (Japan); Urata, K.; Kudough, F. [Mitsubishi Heavy Industries, Ltd., Tokyo (Japan); Hasegawa, M.; Oyabu, I. [Mitsubishi Electric Co., Tokyo (Japan); Yagi, Y.; Sekine, S.; Shimada, T.; Hirano, Y.; Sakakita, H.; Koguchi, H. [Electrotechnical Laboratory, Tsukuba (Japan)

1999-10-01

TPE-RX is a newly constructed, large-sized reversed field pinch (RFP) machine installed at the Electrotechnical Laboratory of the Ministry of International Trade and Industry in Japan. This is the third largest RFP in the world. Major and minor radii of the plasma are 1.72 and 0.45 m, respectively. TPE-RX aims to optimize plasma confinement up to 1 MA. RFP plasma configuration was successfully obtained in March 1998. This paper reports the structural design and manufacturing of the vacuum vessel of TPE-RX. The supporting system on the bellows sections of the vessel was designed based on a detailed finite element method. The integrity of the vacuum vessel against a plasma disruption has been confirmed using dynamic inelastic analyses. (orig.)

Structural design and manufacturing of TPE-RX vacuum vessel

International Nuclear Information System (INIS)

Sago, H.; Orita, J.; Kaguchi, H.; Ishigami, Y.; Urata, K.; Kudough, F.; Hasegawa, M.; Oyabu, I.; Yagi, Y.; Sekine, S.; Shimada, T.; Hirano, Y.; Sakakita, H.; Koguchi, H.

1999-01-01

TPE-RX is a newly constructed, large-sized reversed field pinch (RFP) machine installed at the Electrotechnical Laboratory of the Ministry of International Trade and Industry in Japan. This is the third largest RFP in the world. Major and minor radii of the plasma are 1.72 and 0.45 m, respectively. TPE-RX aims to optimize plasma confinement up to 1 MA. RFP plasma configuration was successfully obtained in March 1998. This paper reports the structural design and manufacturing of the vacuum vessel of TPE-RX. The supporting system on the bellows sections of the vessel was designed based on a detailed finite element method. The integrity of the vacuum vessel against a plasma disruption has been confirmed using dynamic inelastic analyses. (orig.)

The implementation of a data acquisition and service system based on HDF5

Energy Technology Data Exchange (ETDEWEB)

Chen, Y., E-mail: cheny@ipp.ac.cn [Institute of Plasma Physics, Chinese Academy of Sciences, Hefei, Anhui (China); Wang, F.; Li, S. [Institute of Plasma Physics, Chinese Academy of Sciences, Hefei, Anhui (China); Xiao, B.J. [Institute of Plasma Physics, Chinese Academy of Sciences, Hefei, Anhui (China); School of nuclear science and technology, University of Science and Technology of China, Hefei, Anhui (China); Yang, F. [Institute of Plasma Physics, Chinese Academy of Sciences, Hefei, Anhui (China); Department of Computer Science, Anhui Medical University, Hefei, Anhui (China)

2016-11-15

Highlights: • A new data acquisition and service system has been designed and implemented for a new reversed field pinch (RFP) magnetic confinement device. • The new data acquisition and service system is based on HDF5. • It is an entire system including acquisition, storage and data retrieval. • The system is easy to extend and maintain for its modularization design. - Abstract: A data acquisition and service system based on HDF5 has been designed. It includes four components: data acquisition console, data acquisition subsystem, data archive system and data service. The data acquisition console manages all DAQ information and controls the acquisition process. The data acquisition subsystem supports continuous data acquisition with different sampling rates which can be divided into low, medium and high level. All experimental data will be remotely transferred to the data archive system. It adopts HDF5 as its low-level data storage format. The hierarchical data structure of HDF5 is useful for efficiently managing the experimental data and allows users to define special data types and compression filter which can be useful to deal with special signals. Several data service tools have also been developed so that users can get data service via Client/Server or Brower/Server. The system will be demonstrated on Keda Torus eXperiment (KTX) device, which is a new Reversed Field Pinch (RFP) magnetic confinement device. The details are presented in the paper.

The implementation of a data acquisition and service system based on HDF5

International Nuclear Information System (INIS)

Chen, Y.; Wang, F.; Li, S.; Xiao, B.J.; Yang, F.

2016-01-01

Highlights: • A new data acquisition and service system has been designed and implemented for a new reversed field pinch (RFP) magnetic confinement device. • The new data acquisition and service system is based on HDF5. • It is an entire system including acquisition, storage and data retrieval. • The system is easy to extend and maintain for its modularization design. - Abstract: A data acquisition and service system based on HDF5 has been designed. It includes four components: data acquisition console, data acquisition subsystem, data archive system and data service. The data acquisition console manages all DAQ information and controls the acquisition process. The data acquisition subsystem supports continuous data acquisition with different sampling rates which can be divided into low, medium and high level. All experimental data will be remotely transferred to the data archive system. It adopts HDF5 as its low-level data storage format. The hierarchical data structure of HDF5 is useful for efficiently managing the experimental data and allows users to define special data types and compression filter which can be useful to deal with special signals. Several data service tools have also been developed so that users can get data service via Client/Server or Brower/Server. The system will be demonstrated on Keda Torus eXperiment (KTX) device, which is a new Reversed Field Pinch (RFP) magnetic confinement device. The details are presented in the paper.

Quinoline derivative containing monomeric and polymeric metal carboxylates: Synthesis, crystal structure and gas adsorption study over a 2D layered framework

Science.gov (United States)

Gayen, Saikat; Saha, Debraj; Koner, Subratanath

2018-06-01

A new supramolecular metal-carboxylate framework [Co(mqc)2]n (1), and another monomeric compound [Zn (mqc)2(H2O)] (2) (mqcH = 4-methoxy 2-quinolinecarboxylic acid) have been synthesized solvothermally and characterized by single crystal X-ray diffraction, elemental analysis, IR spectra, UV-vis spectra, powdered X-ray diffraction (PXRD) and thermogravimetric analysis. Compound 1 is a 2D coordination polymer, extended to a 3D porous supramolecular network having void space in between 2D layers. Compound 1 exhibits gas uptake capacity of N2, H2, CO2 and CH4 like small gas molecules in which moderately high uptake of H2 and CO2 takes place among the 2D MOFs. While the Zn variety, compound 2 features a one-dimensional chain like structure through strong intermolecular hydrogen-bonding.

Model of a DNA-protein complex of the architectural monomeric protein MC1 from Euryarchaea.

Directory of Open Access Journals (Sweden)

Françoise Paquet

Full Text Available In Archaea the two major modes of DNA packaging are wrapping by histone proteins or bending by architectural non-histone proteins. To supplement our knowledge about the binding mode of the different DNA-bending proteins observed across the three domains of life, we present here the first model of a complex in which the monomeric Methanogen Chromosomal protein 1 (MC1 from Euryarchaea binds to the concave side of a strongly bent DNA. In laboratory growth conditions MC1 is the most abundant architectural protein present in Methanosarcina thermophila CHTI55. Like most proteins that strongly bend DNA, MC1 is known to bind in the minor groove. Interaction areas for MC1 and DNA were mapped by Nuclear Magnetic Resonance (NMR data. The polarity of protein binding was determined using paramagnetic probes attached to the DNA. The first structural model of the DNA-MC1 complex we propose here was obtained by two complementary docking approaches and is in good agreement with the experimental data previously provided by electron microscopy and biochemistry. Residues essential to DNA-binding and -bending were highlighted and confirmed by site-directed mutagenesis. It was found that the Arg25 side-chain was essential to neutralize the negative charge of two phosphates that come very close in response to a dramatic curvature of the DNA.

A review of the experimental and theoretical status of the reversed-field pinch

International Nuclear Information System (INIS)

Baker, D.A.

1987-01-01

This paper reviews the status of the reversed-field pinch (RFP) approach to the development of a compact nuclear fusion reactor. Two RFP papers in this conference are complementary; the first paper contains the historical origins and basic concepts concerning MHD instabilities, relaxation and RFP confinement properties as well as a discussion of future prospects of the RFP. This paper gives an overview of the status of plasma parameters of the present main RFP experiments and of the status of theory and experiment of the interesting RFP plasma phenomena of relaxation, self reversal and flux generation (these effects are often referred to as the dynamo effect). The low frequency oscillating-field current drive concept which exploits these effects is discussed. Particular emphasis is given to the theoretical results obtained from plasma simulation codes used in these active areas of study. Selected topics of recent research on the Los Alamos ZT-40M experiments are reported. The paper concludes with descriptions of the next generation Los Alamos RFP experiment ZTH, to be located in the new Confinement Physics Research Facility (CPRF) presently under construction, and the characteristics of an RFP compact reactor. 68 refs

Pleiotropic benefit of monomeric and oligomeric flavanols on vascular health--a randomized controlled clinical pilot study.

Directory of Open Access Journals (Sweden)

Antje R Weseler

Full Text Available BACKGROUND: Cardiovascular diseases are expanding to a major social-economic burden in the Western World and undermine man's deep desire for healthy ageing. Epidemiological studies suggest that flavanol-rich foods (e.g. grapes, wine, chocolate sustain cardiovascular health. For an evidenced-based application, however, sound clinical data on their efficacy are strongly demanded. METHODS: In a double-blind, randomized, placebo-controlled intervention study we supplemented 28 male smokers with 200 mg per day of monomeric and oligomeric flavanols (MOF from grape seeds. At baseline, after 4 and 8 weeks we measured macro- and microvascular function and a cluster of systemic biomarkers for major pathological processes occurring in the vasculature: disturbances in lipid metabolism and cellular redox balance, and activation of inflammatory cells and platelets. RESULTS: In the MOF group serum total cholesterol and LDL decreased significantly (P ≤ 0.05 by 5% (n = 11 and 7% (n = 9, respectively in volunteers with elevated baseline levels. Additionally, after 8 weeks the ratio of glutathione to glutathione disulphide in erythrocytes rose from baseline by 22% (n = 15, P<0.05 in MOF supplemented subjects. We also observed that MOF supplementation exerts anti-inflammatory effects in blood towards ex vivo added bacterial endotoxin and significantly reduces expression of inflammatory genes in leukocytes. Conversely, alterations in macro- and microvascular function, platelet aggregation, plasma levels of nitric oxide surrogates, endothelin-1, C-reactive protein, fibrinogen, prostaglandin F2alpha, plasma antioxidant capacity and gene expression levels of antioxidant defense enzymes did not reach statistical significance after 8 weeks MOF supplementation. However, integrating all measured effects into a global, so-called vascular health index revealed a significant improvement of overall vascular health by MOF compared to placebo (P ≤ 0.05. CONCLUSION: Our

Does shaping bring an advantage for reversed field pinch plasmas?

International Nuclear Information System (INIS)

Guo, S.C.; Xu, X.Y.; Wang, Z.R.; Liu, Y.Q.

2013-01-01

The MHD–kinetic hybrid toroidal stability code MARS-K (Liu et al 2008 Phys. Plasmas 15 112503) is applied to study the shaping effects on magnetohydrodynamic (MHD) stabilities in reversed field pinch (RFP) plasmas, where both elongation and triangularity are taken into account. The ideal wall β (the ratio of the gaso-kinetic to magnetic pressures) limit set by the ideal kink mode/resistive wall mode in shaped RFP is investigated first, followed by a study of the kinetic damping on the resistive wall mode. Physics understanding of the results is provided by a systematic numerical analysis. Furthermore, the stability boundary of the linear resistive tearing mode in shaped RFP plasmas is computed and compared with that of the circular case. Finally, bootstrap currents are calculated for both circular and shaped RFP plasmas. Overall, the results of these studies indicate that the current circular cross-section is an appropriate choice for RFP devices, in the sense that the plasma shaping does not bring an appreciable advantage to the RFP performance in terms of macroscopic stabilities. In order to reach a steady-state operation, future RFP fusion reactors will probably need a substantial fraction of external current drives, due to the unfavourable scaling for the plasma-generated bootstrap current in the RFP configuration. (paper)

Controlled catalytic and thermal sequential pyrolysis and hydrolysis of polycarbonate and plastic waste to recover monomers

Science.gov (United States)

Evans, R.J.; Chum, H.L.

1994-06-14

A process is described using fast pyrolysis to convert a plastic waste feed stream containing polycarbonate and ABS to high value monomeric constituents prior to pyrolysis of other plastic components therein comprising: selecting a first temperature program range to cause pyrolysis of a given polymer to its high value monomeric constituents prior to a temperature range that causes pyrolysis of other plastic components; selecting an acid or base catalysts and an oxide or carbonate support for treating the feed stream to affect acid or base catalyzed reaction pathways to maximize yield or enhance separation of the high value monomeric constituents of polycarbonate and ABS in the first temperature program range; differentially heating the feed stream at a heat rate within the first temperature program range to provide differential pyrolysis for selective recovery of optimum quantities of the high value monomeric constituents prior to pyrolysis or other plastic components; separating the high value monomeric constituents from the polycarbonate to cause pyrolysis to a different high value monomeric constituent of the plastic waste and differentially heating the feed stream at the second higher temperature program range to cause pyrolysis of different high value monomeric constituents; and separating the different high value monomeric constituents. 68 figs.

Controlled catalystic and thermal sequential pyrolysis and hydrolysis of polycarbonate and plastic waste to recover monomers

Science.gov (United States)

Evans, Robert J.; Chum, Helena L.

1994-01-01

A process of using fast pyrolysis to convert a plastic waste feed stream containing polycarbonate and ABS to high value monomeric constituents prior to pyrolysis of other plastic components therein comprising: selecting a first temperature program range to cause pyrolysis of a given polymer to its high value monomeric constituents prior to a temperature range that causes pyrolysis of other plastic components; selecting an acid or base catalysts and an oxide or carbonate support for treating the feed stream to affect acid or base catalyzed reaction pathways to maximize yield or enhance separation of the high value monomeric constituents of polycarbonate and ABS in the first temperature program range; differentially heating the feed stream at a heat rate within the first temperature program range to provide differential pyrolysis for selective recovery of optimum quantities of the high value monomeric constituents prior to pyrolysis or other plastic components; separating the high value monomeric constituents from the polycarbonate to cause pyrolysis to a different high value monomeric constituent of the plastic waste and differentially heating the feed stream at the second higher temperature program range to cause pyrolysis of different high value monomeric constituents; and separating the different high value monomeric constituents.

Confinement dynamics in the reversed field pinch

International Nuclear Information System (INIS)

Schoenberg, K.F.

1988-01-01

The study of basic transport and confinement dynamics is central to the development of the reversed field pinch (RFP) as a confinement concept. Thus, the goal of RFP research is to understand the connection between processes that sustain the RFP configuration and related transport/confinement properties. Recently, new insights into confinement have emerged from a detailed investigation of RFP electron and ion physics. These insights derive from the recognition that both magnetohydrodynamic (MHD) and electron kinetic effects play an important and strongly coupled role in RFP sustainment and confinement dynamics. In this paper, we summarize the results of these studies on the ZT-40M experiment. 8 refs

Corrosion inhibition of aluminum with a series of aniline monomeric surfactants and their analog polymers in 0.5 M HCl solution

Directory of Open Access Journals (Sweden)

M.M. El-Deeb

2015-07-01

Full Text Available The inhibition effect of 3-(12-sodiumsulfonate dodecyloxy aniline monomeric surfactant (MC12 and its analog polymer Poly 3-(dodecyloxy sulfonic acid aniline (PC12 on the corrosion of aluminum in 0.5 M HCl solution was investigated using weight loss and potentiodynamic polarization techniques. The presence of these two compounds in 0.5 M HCl inhibits the corrosion of aluminum without modifying the mechanism of corrosion process. It was found that these inhibitors act as mixed-type inhibitors with anodic predominance as well as the inhibition efficiency increases with increasing inhibitor concentration, but decreases with raising temperature. Langmuir and Frumkin adsorption isotherms fit well with the experimental data. Thermodynamic functions for both dissolution and adsorption processes were determined. The obtained results from weight loss and potentiodynamic polarization techniques are in good agreement with contact angle measurements.

Analysis of monomeric and oligomeric organophosphorus flame retardants in fish muscle tissues using liquid chromatography–electrospray ionization tandem mass spectrometry: Application to Nile tilapia (Oreochromis niloticus) from an e-waste processing area in northern Vietnam

OpenAIRE

Matsukami, Hidenori; Suzuki, Go; Tue, Nguyen Minh; Tuyen, Le Huu; Viet, Pham Hung; Takahashi, Shin; Tanabe, Shinsuke; Takigami, Hidetaka

2016-01-01

Using electrospray ionization tandem mass spectrometry combined with liquid chromatography (LC), a novel analytical method was developed to quantify eight monomeric organophosphorus flame retardants (m-PFRs) and three oligomeric organophosphorus flame retardants (o-PFRs) in fish muscle samples. The optimization and validation experiments indicate that the developed method can determine accurately the concentrations of analytes in fish muscle samples. The recoveries of analytes in fish muscle ...

Effect of artificial accelerated aging on the optical properties and monomeric conversion of composites used after expiration date.

Science.gov (United States)

Garcia, Lucas da Fonseca Roberti; Mundim, Fabricio Mariano; Pires-de-Souza, Fernanda de Carvalho Panzeri; Puppin Rontani, Regina Maria; Consani, Simonides

2013-01-01

This study sought to evaluate how artificial accelerated aging (AAA) affected color stability (ΔE), opacity (ΔOP), and degree of conversion (DOC) for 3 composite materials (Tetric Ceram, Tetric Ceram HB, and Tetric Flow) used both 180 days before and 180 days after their expiration dates. To evaluate the materials' optical properties, 10 specimens of each composite-5 prior to expiration and 5 after the materials' expiration date-were made in a teflon matrix. After polishing, the specimens were submitted to initial color and opacity readings and submitted to AAA for 384 hours; at that point, new readings were taken to determine ΔE and ΔOP. To evaluate monomeric conversion evaluation, 6 specimens from each composite and expiration date-3 prior to AAA and 3 after-were submitted to DOC analysis. Results of the 2-way ANOVA and Bonferroni's tests (P 0.05). The expired Tetric Flow had the highest DOC values (71.42% ± 4.21) before AAA, significantly different than that of the other composites (P > 0.05). It was concluded that both expiration date and AAA affected the properties of the composites tested.

Optimized labeling of membrane proteins for applications to super-resolution imaging in confined cellular environments using monomeric streptavidin.

Science.gov (United States)

Chamma, Ingrid; Rossier, Olivier; Giannone, Grégory; Thoumine, Olivier; Sainlos, Matthieu

2017-04-01

Recent progress in super-resolution imaging (SRI) has created a strong need to improve protein labeling with probes of small size that minimize the target-to-label distance, increase labeling density, and efficiently penetrate thick biological tissues. This protocol describes a method for labeling genetically modified proteins incorporating a small biotin acceptor peptide with a 3-nm fluorescent probe, monomeric streptavidin. We show how to express, purify, and conjugate the probe to organic dyes with different fluorescent properties, and how to label selectively biotinylated membrane proteins for SRI techniques (point accumulation in nanoscale topography (PAINT), stimulated emission depletion (STED), stochastic optical reconstruction microscopy (STORM)). This method is complementary to the previously described anti-GFP-nanobody/SNAP-tag strategies, with the main advantage being that it requires only a short 15-amino-acid tag, and can thus be used with proteins resistant to fusion with large tags and for multicolor imaging. The protocol requires standard molecular biology/biochemistry equipment, making it easily accessible for laboratories with only basic skills in cell biology and biochemistry. The production/purification/conjugation steps take ∼5 d, and labeling takes a few minutes to an hour.

Red fluorescence of dental plaque in children -A cross-sectional study.

Science.gov (United States)

Volgenant, Catherine M C; Zaura, Egija; Brandt, Bernd W; Buijs, Mark J; Tellez, Marisol; Malik, Gayatri; Ismail, Amid I; Ten Cate, Jacob M; van der Veen, Monique H

2017-03-01

The relation between the presence of red fluorescent plaque and the caries status in children was studied. In addition, the microbial composition of dental plaque from sites with red fluorescent plaque (RFP) and from sites with no red fluorescent plaque (NFP) was assessed. Fluorescence photographs were taken from fifty children (6-14 years old) with overnight plaque. Full-mouth caries scores (ICDAS II) were obtained. The composition of a saliva sample and two plaque samples (RFP and NFP) was assessed using 16S rDNA sequencing. At the site level, no clinically relevant correlations were found between the presence of RFP and the caries status. At the subject level, a weak correlation was found between RFP and the caries status when non-cavitated lesions were included (r s =0.37, p=0.007). The microbial composition of RFP differed significantly from NFP. RFP had more anaerobes and more Gram-negative bacterial taxa. The most discriminative operational taxonomic units (OTUs) for RFP were Corynebacterium, Leptotrichia, Porphyromonas and Selenomonas, while the most discriminative OTUs for NFP were Neisseria, Actinomyces, Streptococcus and Rothia. There were no clinical relevant correlations in this cross-sectional study between the presence of RFP and (early) caries lesions. There were differences in the composition of these phenotypically different plaque samples: RFP contained more Gram-negative, anaerobic taxa and was more diverse than NFP. The study outcomes provide more insight in the possibilities to use plaque fluorescence in oral health risk assessments. Copyright © 2017 Elsevier Ltd. All rights reserved.

Recommended plutonium release fractions from postulated fires. Final report

International Nuclear Information System (INIS)

Kogan, V.; Schumacher, P.M.

1993-12-01

This report was written at the request of EG ampersand G Rocky Flats, Inc. in support of joint emergency planning for the Rocky Flats Plant (RFP) by EG ampersand G and the State of Colorado. The intent of the report is to provide the State of Colorado with an independent assessment of any respirable plutonium releases that might occur in the event of a severe fire at the plant. Fire releases of plutonium are of interest because they have been used by EG ampersand G to determine the RFP emergency planning zones. These zones are based on the maximum credible accident (MCA) described in the RFP Final Environmental Impact Statement (FEIS) of 1980, that MCA is assumed to be a large airplane crashing into a RFP plutonium building.The objective of this report was first, to perform a worldwide literature review of relevant release experiments from 1960 to the present and to summarize those findings, and second, to provide recommendations for application of the experimental data to fire release analyses at Rocky Flats. The latter step requires translation between experimental and expected RFP accident parameters, or ''scaling.'' The parameters of particular concern are: quantities of material, environmental parameters such as the intensity of a fire, and the physico-chemical forms of the plutonium. The latter include plutonium metal, bulk plutonium oxide powder, combustible and noncombustible wastes contaminated with plutonium oxide powder, and residues from plutonium extraction processes

Responding to an RFP: A Vendor's Viewpoint.

Science.gov (United States)

Kington, Robert A.

1987-01-01

Outlines factors used by online vendors to decide whether to bid on RFPs (requests for proposals) for library automation systems, including specifications for software, hardware or performance requirements not met by the vendor; specifications based on competitors' systems; the size and complexity of the request itself; and vendors' time…

Hydrolyzable tannins of tamaricaceous plants. V. Structures of monomeric-trimeric tannins and cytotoxicity of macrocyclic-type tannins isolated from Tamarix nilotica (1).

Science.gov (United States)

Orabi, Mohamed A A; Taniguchi, Shoko; Sakagami, Hiroshi; Yoshimura, Morio; Yoshida, Takashi; Hatano, Tsutomu

2013-05-24

Three new ellagitannin monomers, nilotinins M5-M7 (1-3), a dimer, nilotinin D10 (4), and a trimer, nilotinin T1 (5), together with three known dimers, hirtellin D (7) and tamarixinins B (8) and C (9), and a trimer, hirtellin T2 (6), were isolated from Tamarix nilotica dried leaves. The structures of the tannins were elucidated by intensive spectroscopic methods and chemical conversions into known tannins. The new trimer (5) is a unique macrocyclic type whose monomeric units are linked together by an isodehydrodigalloyl and two dehydrodigalloyl moieties. Additionally, dimeric and trimeric macrocyclic-type tannins isolated from T. nilotica in this study were assessed for possible cytotoxic activity against four human tumor cell lines. Tumor-selective cytotoxicities of the tested compounds were higher than those of synthetic and natural potent cytotoxic compounds, including polyphenols, and comparable with those of 5-fluorouracil and melphalan.

Behaviour of the peripheral plasma in the reversed field pinch

International Nuclear Information System (INIS)

Matsuoka, A.; Sato, K.I.; Arimoto, H.; Yamada, S.; Nagata, A.; Murata, H.

1986-01-01

By using Langmuir probes installed behind limiters, time behaviour of the peripheral plasma in the Reversed Field Pinch (RFP) are investigated. They are strongly affected by the confined RFP plasma and are divided into three phases (the initial phase before setting up the RFP configuration, the current rising phase, and the quiescent phase), which are just the same as those of the confined RFP plasma. Typical behaviour of the peripheral plasma have relations to the pump out phenomena and of the toroidal flux generation. (author)

Expression pattern of Ccr2 and Cx3cr1 in inherited retinal degeneration.

Science.gov (United States)

Kohno, Hideo; Koso, Hideto; Okano, Kiichiro; Sundermeier, Thomas R; Saito, Saburo; Watanabe, Sumiko; Tsuneoka, Hiroshi; Sakai, Tsutomu

2015-10-12

Though accumulating evidence suggests that microglia, resident macrophages in the retina, and bone marrow-derived macrophages can cause retinal inflammation which accelerates photoreceptor cell death, the details of how these cells are activated during retinal degeneration (RD) remain uncertain. Therefore, it is important to clarify which cells play a dominant role in fueling retinal inflammation. However, distinguishing between microglia and macrophages is difficult using conventional techniques such as cell markers (e.g., Iba-1). Recently, two mouse models for visualizing chemokine receptors were established, Cx3cr1 (GFP/GFP) and Ccr2 (RFP/RFP) mice. As Cx3cr1 is expressed in microglia and Ccr2 is reportedly expressed in activated macrophages, these mice have the potential to distinguish microglia and macrophages, yielding novel information about the activation of these inflammatory cells and their individual roles in retinal inflammation. In this study, c-mer proto-oncogene tyrosine kinase (Mertk) (-/-) mice, which show photoreceptor cell death due to defective retinal pigment epithelium phagocytosis, were employed as an animal model of RD. Mertk (-/-) Cx3cr1 (GFP/+) Ccr2 (RFP/+) mice were established by breeding Mertk (-/-) , Cx3cr1 (GFP/GFP) , and Ccr2 (RFP/RFP) mice. The retinal morphology and pattern of inflammatory cell activation and invasion of Mertk (-/-) Cx3cr1 (GFP/+) Ccr2 (RFP/+) mice were evaluated using retina and retinal pigment epithelium (RPE) flat mounts, retinal sections, and flow cytometry. Four-week-old Mertk (-/-) Cx3cr1 (GFP/+) Ccr2 (RFP/+) mice showed Cx3cr1-GFP-positive microglia in the inner retina. Cx3cr1-GFP and Ccr2-RFP dual positive activated microglia were observed in the outer retina and subretinal space of 6- and 8-week-old animals. Ccr2-RFP single positive bone marrow-derived macrophages were observed to migrate into the retina of Mertk (-/-) Cx3cr1 (GFP/+) Ccr2 (RFP/+) mice. These invading cells were still observed in the

The monomeric form of Neisseria DNA mimic protein DMP19 prevents DNA from binding to the histone-like HU protein

Science.gov (United States)

Ko, Tzu-Ping; Liao, Yi-Ting; Hsu, Kai-Cheng

2017-01-01

DNA mimicry is a direct and effective strategy by which the mimic competes with DNA for the DNA binding sites on other proteins. Until now, only about a dozen proteins have been shown to function via this strategy, including the DNA mimic protein DMP19 from Neisseria meningitides. We have shown previously that DMP19 dimer prevents the operator DNA from binding to the transcription factor NHTF. Here, we provide new evidence that DMP19 monomer can also interact with the Neisseria nucleoid-associated protein HU. Using BS3 crosslinking, gel filtration and isothermal titration calorimetry assays, we found that DMP19 uses its monomeric form to interact with the Neisseria HU dimer. Crosslinking conjugated mass spectrometry was used to investigate the binding mode of DMP19 monomer and HU dimer. Finally, an electrophoretic mobility shift assay (EMSA) confirmed that the DNA binding affinity of HU is affected by DMP19. These results showed that DMP19 is bifunctional in the gene regulation of Neisseria through its variable oligomeric forms. PMID:29220372

A 12-week DBPC dose-finding study with sublingual monomeric allergoid tablets in house dust mite-allergic patients.

Science.gov (United States)

Hüser, C; Dieterich, P; Singh, J; Shah-Hosseini, K; Allekotte, S; Lehmacher, W; Compalati, E; Mösges, R

2017-01-01

In sublingual immunotherapy, optimal doses are a key factor for therapeutic outcomes. The aim of this study with tablets containing carbamylated monomeric house dust mite allergoids was to determine the most effective and safe dose. In this double-blind, placebo-controlled dose-finding study, 131 patients with house dust mite-induced allergic rhinoconjunctivitis were randomized to 12-week treatments with 300 UA/day, 1000 UA/day, 2000 UA/day, 3000 UA/day or placebo. Conjunctival provocation tests (CPT) were performed before, during and after treatment. The change in mean allergic severity (primary endpoint), calculated from the severity of the CPT reaction, and the proportion of patients with an improved CPT threshold (secondary endpoint) determined the treatment effect. The mean allergic severity decreased in all groups, including the placebo group. It was lower in all active treatment groups (300 UA/day: 0.14, 1000 UA/day: 0.15, 2000 UA/day: 0.10, 3000 UA/day: 0.15) than in the placebo group (0.30). However, this difference was not statistically significant (P allergoid sublingual tablets is well tolerated and reduces the CPT reaction in house dust mite-allergic patients. © 2016 The Authors. Allergy Published by John Wiley & Sons Ltd.

Monomeric, porous type II collagen scaffolds promote chondrogenic differentiation of human bone marrow mesenchymal stem cells in vitro

Science.gov (United States)

Tamaddon, M.; Burrows, M.; Ferreira, S. A.; Dazzi, F.; Apperley, J. F.; Bradshaw, A.; Brand, D. D.; Czernuszka, J.; Gentleman, E.

2017-03-01

Osteoarthritis (OA) is a common cause of pain and disability and is often associated with the degeneration of articular cartilage. Lesions to the articular surface, which are thought to progress to OA, have the potential to be repaired using tissue engineering strategies; however, it remains challenging to instruct cell differentiation within a scaffold to produce tissue with appropriate structural, chemical and mechanical properties. We aimed to address this by driving progenitor cells to adopt a chondrogenic phenotype through the tailoring of scaffold composition and physical properties. Monomeric type-I and type-II collagen scaffolds, which avoid potential immunogenicity associated with fibrillar collagens, were fabricated with and without chondroitin sulfate (CS) and their ability to stimulate the chondrogenic differentiation of human bone marrow-derived mesenchymal stem cells was assessed. Immunohistochemical analyses showed that cells produced abundant collagen type-II on type-II scaffolds and collagen type-I on type-I scaffolds. Gene expression analyses indicated that the addition of CS - which was released from scaffolds quickly - significantly upregulated expression of type II collagen, compared to type-I and pure type-II scaffolds. We conclude that collagen type-II and CS can be used to promote a more chondrogenic phenotype in the absence of growth factors, potentially providing an eventual therapy to prevent OA.

Advanced concepts in the United States fusion program

International Nuclear Information System (INIS)

Dove, W.F.

1985-01-01

The goal of the magnetic fusion program is to establish the scientific and technological base for fusion energy. Development of a variety of magnetic confinement systems is essential to achieving that goal. The role of the advanced concepts program is to conduct experimental investigations of confinement concepts other than the tokamaks and tandem mirror concepts. The present advanced concepts program consists of the reversed-field-pinch (RFP), the spheromak and the field-reversed configuration (FRC). Significant new experiments in the RFP and FRC concepts have been approved and are described

A program of FRC theory research

International Nuclear Information System (INIS)

Krall, N.A.

1990-01-01

At the request of the Office of Fusion Energy, a group of experts was convened on February 6--8, 1990. This group met to assess the world data base on Reversed Field Pinch (RFP) physics, and, further, to assess the role of the ZTH experiment in providing reactor relevant physics understanding for that confinement geometry. This group met, analyzed some of the relevant literature, and heard extensive presentations on the physics of the RFP and the plans for the ZTH and RFX devices. The conclusions of this group of experts are contained in this report

Reversed field pinch ignition requirements

International Nuclear Information System (INIS)

Werley, K.A.

1991-01-01

Plasma models are described and used to calculated numerically the transport confinement (nτ E ) requirements and steady state operation points for both the reversed field pinch (RFP) and the tokamak. The models are used to examine the CIT tokamak ignition conditions and the RFP experimental and ignition conditions. Physics differences between RFPs and tokamaks and their consequences for a D-T ignition machine are discussed. Compared with a tokamak, the ignition RFP has many physics advantages, including Ohmic heating to ignition (no need for auxiliary heating systems), higher beta, lower ignition current, less sensitivity of ignition requirements to impurity effects, no hard disruptions (associated with beta or density limits) and successful operation with high radiation fractions (f RAD ∼ 0.95). These physics advantages, coupled with important engineering advantages associated with lower external magnetic field, larger aspect ratios and smaller plasma cross-sections, translate to significant cost reductions for both ignition and reactor applications. The primary drawback of the RFP is the uncertainty that the present scaling will extrapolate to reactor regimes. Devices that are under construction should go a long way toward resolving this scaling uncertainty. The 4 MA ZTH is expected to extend the nτ E transport scaling data by three orders of magnitude above the results of ZT-40M, and, if the present scaling holds, ZTH is expected to achieve a D-T equivalent scientific energy breakeven, Q = 1. A base case RFP ignition point is identified with a plasma current of 8.1 MA and no auxiliary heating. (author). 19 refs, 11 figs, 3 tabs

Expression of membrane-associated proteins within single emulsion cell facsimiles.

Science.gov (United States)

Chanasakulniyom, Mayuree; Martino, Chiara; Paterson, David; Horsfall, Louise; Rosser, Susan; Cooper, Jonathan M

2012-07-07

MreB is a structural membrane-associated protein which is one of the key components of the bacterial cytoskeleton. Although it plays an important role in shape maintenance of rod-like bacteria, the understanding of its mechanism of action is still not fully understood. This study shows how segmented flow and microdroplet technology can be used as a new tool for biological in vitro investigation of this protein. In this paper, we demonstrate cell-free expression in a single emulsion system to express red fluorescence protein (RFP) and MreB linked RFP (MreB-RFP). We follow the aggregation and localisation of the fusion protein MreB-RFP in this artificial cell-like environment. The expression of MreB-RFP in single emulsion droplets leads to the formation of micrometer-scale protein patches distributed at the water/oil interface.

EPA's information management system for tracking reasonable further progress. Report for October 1992-May 1993

Energy Technology Data Exchange (ETDEWEB)

Barnard, W.R.; Monroe, C.C.; Dean, T.A.

1993-01-01

The paper describes operating characteristics of a personal-computer (PC)-based Reasonable Further Progress (RFP) tracking system developed for use by EPA to evaluate the progress that nonattainment areas are making toward meeting a 15% volatile organic compound (VOC) reduction specified in the Clean Air Act Amendments (CAAA) of 1990. Details of the program's capabilities, file handling, reporting, and graphics are reported. In order to track the emission reductions resulting from these strategies, emissions reported in the RFP projection inventories will be compared with actual emissions reported in periodic adjusted-base inventories, that are also required by the CAAA. However, the comparison will take place a number of years in the future.

Proceedings of the international workshop on engineering design of next step reversed field pinch devices

International Nuclear Information System (INIS)

Thomson, D.B.

1987-11-01

These Proceedings contain the formal contributed papers, the workshop papers and workshop summaries presented at the International Workshop on Engineering Design of Next Step RFP Devices held at Los Alamos, July 13-17, 1987. Contributed papers were presented at formal sessions on the topics: (1) physics overview (3 papers); (2) general overview (3 papers); (3) front-end (9 papers); (4) computer control and data acquisition (1 paper); (5) magnetics (5 papers); and (6) electrical design (9 papers). Informal topical workshop sessions were held on the topics: (1) RFP physics (9 papers); (2) front-end (7 papers); (3) magnetics (3 papers); and (4) electrical design (1 paper). This volume contains the summaries written by the Chairmen of each of the informal topical workshop sessions. The papers in these Proceedings represent a significant review of the status of the technical base for the engineering design of the next step RFP devices being developed in the US, Europe, and Japan, as of this date

Proceedings of the international workshop on engineering design of next step reversed field pinch devices

Energy Technology Data Exchange (ETDEWEB)

Thomson, D.B. (comp.)

1987-11-01

These Proceedings contain the formal contributed papers, the workshop papers and workshop summaries presented at the International Workshop on Engineering Design of Next Step RFP Devices held at Los Alamos, July 13-17, 1987. Contributed papers were presented at formal sessions on the topics: (1) physics overview (3 papers); (2) general overview (3 papers); (3) front-end (9 papers); (4) computer control and data acquisition (1 paper); (5) magnetics (5 papers); and (6) electrical design (9 papers). Informal topical workshop sessions were held on the topics: (1) RFP physics (9 papers); (2) front-end (7 papers); (3) magnetics (3 papers); and (4) electrical design (1 paper). This volume contains the summaries written by the Chairmen of each of the informal topical workshop sessions. The papers in these Proceedings represent a significant review of the status of the technical base for the engineering design of the next step RFP devices being developed in the US, Europe, and Japan, as of this date.

Total allowable concentrations of monomeric inorganic aluminum and hydrated aluminum silicates in drinking water.

Science.gov (United States)

Willhite, Calvin C; Ball, Gwendolyn L; McLellan, Clifton J

2012-05-01

Maximum contaminant levels are used to control potential health hazards posed by chemicals in drinking water, but no primary national or international limits for aluminum (Al) have been adopted. Given the differences in toxicological profiles, the present evaluation derives total allowable concentrations for certain water-soluble inorganic Al compounds (including chloride, hydroxide, oxide, phosphate and sulfate) and for the hydrated Al silicates (including attapulgite, bentonite/montmorillonite, illite, kaolinite) in drinking water. The chemistry, toxicology and clinical experience with Al materials are extensive and depend upon the particular physical and chemical form. In general, the water solubility of the monomeric Al materials depends on pH and their water solubility and gastrointestinal bioavailability are much greater than that of the hydrated Al silicates. Other than Al-containing antacids and buffered aspirin, food is the primary source of Al exposure for most healthy people. Systemic uptake of Al after ingestion of the monomeric salts is somewhat greater from drinking water (0.28%) than from food (0.1%). Once absorbed, Al accumulates in bone, brain, liver and kidney, with bone as the major site for Al deposition in humans. Oral Al hydroxide is used routinely to bind phosphate salts in the gut to control hyperphosphatemia in people with compromised renal function. Signs of chronic Al toxicity in the musculoskeletal system include a vitamin D-resistant osteomalacia (deranged membranous bone formation characterized by accumulation of the osteoid matrix and reduced mineralization, reduced numbers of osteoblasts and osteoclasts, decreased lamellar and osteoid bands with elevated Al concentrations) presenting as bone pain and proximal myopathy. Aluminum-induced bone disease can progress to stress fractures of the ribs, femur, vertebrae, humerus and metatarsals. Serum Al ≥100 µg/L has a 75-88% positive predictive value for Al bone disease. Chronic Al

Alpha effect of Alfven waves and current drive in reversed field pinches

International Nuclear Information System (INIS)

Litwin, C.; Prager, S.C.

1997-10-01

Circularly polarized Alfven waves give rise to an α-dynamo effect that can be exploited to drive parallel current. In a open-quotes laminarclose quotes magnetic the effect is weak and does not give rise to significant currents for realistic parameters (e.g., in tokamaks). However, in reversed field pinches (RFPs) in which magnetic field in the plasma core is stochastic, a significant enhancement of the α-effect occurs. Estimates of this effect show that it may be a realistic method of current generation in the present-day RFP experiments and possibly also in future RFP-based fusion reactors

Controlled catalytic and thermal sequential pyrolysis and hydrolysis of polymer waste comprising nylon 6 and a polyolefin or mixtures of polyolefins to sequentially recover monomers or other high value products

Science.gov (United States)

Evans, R.J.; Chum, H.L.

1994-10-25

A process of using fast pyrolysis in a carrier gas to convert a plastic waste feedstream having a mixed polymeric composition in a manner such that pyrolysis of a given polymer to its high value monomeric constituent occurs prior to pyrolysis of other plastic components therein comprising: selecting a first temperature program range to cause pyrolysis of said given polymer to its high value monomeric constituent prior to a temperature range that causes pyrolysis of other plastic components; selecting a catalyst and support for treating said feed streams with said catalyst to effect acid or base catalyzed reaction pathways to maximize yield or enhance separation of said high value monomeric constituent in said temperature program range; differentially heating said feed stream at a heat rate within the first temperature program range to provide differential pyrolysis for selective recovery of optimum quantities of the high value monomeric constituent prior to pyrolysis of other plastic components; separating the high value monomeric constituents; selecting a second higher temperature range to cause pyrolysis of a different high value monomeric constituent of said plastic waste and differentially heating the feedstream at the higher temperature program range to cause pyrolysis of the different high value monomeric constituent; and separating the different high value monomeric constituent. 83 figs.

Stability of monomeric Cro variants: Isoenergetic transformation of a type I' to a type II' beta-hairpin by single amino acid replacements.

Science.gov (United States)

Mollah, A K M M; Stennis, Rhonda L; Mossing, Michael C

2003-05-01

The thermodynamic stabilities of three monomeric variants of the bacteriophage lambda Cro repressor that differ only in the sequence of two amino acids at the apex of an engineered beta-hairpin have been determined. The sequences of the turns are EVK-XX-EVK, where the two central residues are DG, GG, and GT, respectively. Standard-state unfolding free energies, determined from circular dichroism measurements as a function of urea concentration, range from 2.4 to 2.7 kcal/mole, while those determined from guanidine hydrochloride range from 2.8 to 3.3 kcal/mole for the three proteins. Thermal denaturation yields van't Hoff unfolding enthalpies of 36 to 40 kcal /mole at midpoint temperatures in the range of 53 to 58 degrees C. Extrapolation of the thermal denaturation free energies with heat capacities of 400 to 600 cal/mole deg gives good agreement with the parameters determined in denaturant titrations. As predicted from statistical surveys of amino acid replacements in beta-hairpins, energetic barriers to transformation from a type I' turn (DG) to a type II' turn (GT) can be quite small.

Engineering and physics of high-power-density, compact, reversed-field-pinch fusion reactors

International Nuclear Information System (INIS)

Najmabadi, F.; Conn, R.W.; Krakowski, R.A.; Schultz, K.R.; Steiner, D.

1989-01-01

The technical feasibility and key developmental issues of compact, high-power-density Reversed-Field-Pinch (RFP) reactors are the primary results of the TITAN RFP reactor study. Two design approaches emerged, TITAN-I and TITAN-II, both of which are steady-state, DT-burning, circa 1000 MWe power reactors. The TITAN designs are physically compact and have a high neutron wall loading of 18 MW m 2 . Detailed analyses indicate that: a) each design is technically feasible; b) attractive features of compact RFP reactors can be realized without sacrificing the safety and environmental potential of fusion; and c) major features of this particular embodiment of the RFP reactor are retained in a design window of neutron wall loading ranging from 10 to 20 MW/m 2 . A major product of the TITAN study is the identification and quantification of major engineering and physics requirements for this class of RFP reactors. These findings are the focus of this paper. (author). 26 refs.; 4 figs.; 1 tab

Temperature evolution in a magnetohydrodynamics simulation of a reversed-field pinch

International Nuclear Information System (INIS)

Onofri, M.; Malara, F.; Veltri, P.

2010-01-01

The temperature evolution in a magnetohydrodynamics (MHD) simulation of a reversed-field pinch (RFP) is investigated including thermal conductivity. For numerical reasons, an isotropic thermal conductivity is used, even though in a RFP plasma the parallel conductivity is much larger than the perpendicular one so that magnetic field lines tend to become isothermal. The system shows alternating multiple helicity states and quasi-single helicity states. Single-helical-axis states are formed when the amplitude of the dominant mode is above a determined threshold, as observed in experiments. The relation between heat transport and magnetic field topology that is observed in RFP experiments cannot be found in the simulation, since thermal conductivity is independent of the magnetic field. This difficulty should be taken into account in the numerical investigation of the RFP dynamics. In this paper, the first description of the temperature evolution in a compressible MHD simulation of a RFP is given.

Role of magnetic reconnection phenomena in the reversed-field pinch

International Nuclear Information System (INIS)

Baker, D.A.

1983-01-01

The reversed-field pinch (RFP), an axisymmetric toroidal magnetic confinement experiment, has physics rich in the area commonly called field line reconnection or merging. This paper reviews the topics where reconnection plays a vital role: (a) RFP formation and the phenomenon of self-reversal, (b) RFP sustainment in which the RFP configuration has been shown to be capable of maintaining itself for times much longer than earlier predictions from classical resistive MHD theory, (c) steady state current drive in which dynamo action and associated reconnection processes give rise to the possibility of sustaining the configuration indefinitely by means of low frequency ac modulation of the toroidal and poloidal magnetic fields, (d) the effects of reconnection on the formation and evolution of the magnetic surfaces which are intimately related to the plasma containment properties. It appears that all phases of the RFP operation are intimately related to the reconnection and field regeneration processes similar to those encountered in space and astrophysics

Training Systems Product Group (TSPG) Training Systems Acquisition (TSA)

Science.gov (United States)

2006-05-23

Ceiling Status • Ceiling Reserve Programs to be Awarded – FY 06 • HC-130P Ogden RFP: May 06 • ATARS Ogden RFP: Apr 06 • F-16 MSAMT FMS WP RFP: TOPS...set-asides • Set up a method for potential teams members to identify one another • Continue to make opportunities and award available to industry • Must streamline D.O. bids and turn tasks quickly

Potential of Diverse Prokaryotic Organisms for Glycerol-based Polyhydroxyalkanoate Production

Directory of Open Access Journals (Sweden)

Martin Koller

2015-06-01

Full Text Available The potential and performance of various Gram-negative, Gram-positive and archaeal wild type microorganisms, and bacterial mixed cultures, as well as the application of genetically engineered strains as whole-cell biocatalysts for glycerol-based polyhydroxyalkanoate production are analyzed and assessed. This encompasses the comparison of growth and polyhydroxyalkanoate accumulation kinetics, thermo-mechanical properties of isolated glycerol-based polyhydroxyalkanoate of different composition on the monomeric level, and the presentation of mathematical models developed to describe glycerol-based polyhydroxyalkanoate production processes. For all these aspects, the article provides a detailed compilation of the contemporary state of knowledge, and gives an outlook to expected future developments.

Reversed field pinch diagnostics

International Nuclear Information System (INIS)

Weber, P.G.

1986-01-01

The Reversed Field Pinch (RFP) is a toroidal, axisymmetric magnetic confinement configuration characterized by a magnetic field configuration in which the toroidal magnetic field is of similar strength to the poloidal field, and is reversed at the edge compared to the center. The RFP routinely operates at high beta, and is a strong candidate for a compact fusion device. Relevant attributes of the configuration will be presented, together with an overview of present and planned experiments and their diagnostics. RFP diagnostics are in many ways similar to those of other magnetic confinement devices (such as tokamaks); these lectures will point out pertinent differences, and will present some diagnostics which provide special insights into unique attributes of the RFP

Confinement dynamics and boundary condition studies in the Reversed Field Pinch

International Nuclear Information System (INIS)

Schoenberg, K.F.; Ingraham, J.C.; Moses, R.W. Jr.

1988-01-01

The study of confinement dynamics, including investigation of the boundary conditions required for plasma sustainment, are central to the development of the Reversed Field Pinch (RFP) concept. Recently, several insights into confinement have emerged from a detailed investigation RFP electron and ion dynamics. These insights derive from the recognition that both magnetohydrodynamic (MHD) and electron kinetic effects play an important and coupled role in RFP stability, sustainment, and confinement. In this paper, we summarize the results of confinement studies on the ZT-40M experiment, and boundary condition studies on the Wisconsin non-circular RFP experiment. A brief description of the newly commissioned Madison Symmetric Torus (MST) is also presented. 28 refs., 3 figs

Targeting BRCAness in Gastric Cancer

Science.gov (United States)

2017-10-01

affected in subsets of these tumors. For example, mutations in BRCA1/2 were found in about 15% of gastric cancer, loss of BRCA1 protein expression...platform   Figure 4: Nuclear-RFP tagged SNU1 cell lines after 6 days in culture . (A) Phase contrast. (B) RFP. (C) Markup image of RFP confluence...Title: Cell Cultures and Xenografts from Esophagogastric, Pancreatic, Colorectal and Neuroendocrine Tumors, IACUC protocol number 10-02-003, Protocol

A Color-coded Imageable Syngeneic Mouse Model of Stromal-cell Recruitment by Metastatic Lymphoma.

Science.gov (United States)

Matsumoto, Takuro; Suetsugu, Atsushi; Shibata, Yuhei; Nakamura, Nobuhiko; Aoki, Hitomi; Kunisada, Takahiro; Tsurumi, Hisashi; Shimizu, Masahito; Hoffman, Robert M

2015-09-01

A syngeneic color-coded imageable lymphoma model has been developed to visualize recruitment of host stromal cells by malignant lymphoma during metastasis. The EL4 cell line was previously derived from a lymphoma induced in a C57/BL6 mouse by 9,10-dimethyl-1,2-benzanthracene. EL4 lymphoma cells expressing red fluorescent protein (EL4-RFP) were initially established. EL4-RFP cells were subsequently injected into the tail vein of C57/BL6-GFP transgenic mice. EL4-RFP metastasis was observed in the lymph nodes of the upper mediastinum and in the liver 28 days after cell injection. Large EL4-RFP liver metastases in C57/BL6-GFP mice contained GFP-expressing stromal cells derived from the host. In addition, EL4-RFP lymphoma metastasis was formed in peri-gastric lymph nodes, which were also enriched in host GFP-expressing cells. Furthermore, EL4-RFP lymphoma cells were also observed in the peripheral blood and bone marrow of C57/BL6-GFP transgenic mice, where they were associated with GFP-expressing host cells. Lymph node, liver and bone marrow metastases were found approximately 4 weeks after transplantation and all RFP-expressing metastases were highly enriched in GFP-expressing host stromal cells. This model of malignant lymphoma can be used to study early tumor development, metastasis, and the role of the stroma, as well as for discovery and evaluation of novel therapeutics for this treatment-resistant disease. Copyright© 2015 International Institute of Anticancer Research (Dr. John G. Delinassios), All rights reserved.

I222 crystal form of despentapeptide (B26-B30) insulin provides new insights into the properties of monomeric insulin.

Science.gov (United States)

Whittingham, Jean L; Youshang, Zhang; Záková, Lenka; Dodson, Eleanor J; Turkenburg, Johan P; Brange, Jens; Dodson, G Guy

2006-05-01

Despentapeptide (des-B26-B30) insulin (DPI), an active modified insulin, has been crystallized in the presence of 20% acetic acid pH 2. A crystal structure analysis to 1.8 A spacing (space group I222) revealed that the DPI molecule, which is unable to make beta-strand interactions for physiological dimer formation and is apparently monomeric in solution, formed an alternative lattice-generated dimer. The formation of this dimer involved interactions between surfaces which included the B9-B19 alpha-helices (usually buried by the dimer-dimer contacts within the native hexamer). The two crystallographically independent molecules within the dimer were essentially identical and were similar in conformation to T-state insulin as seen in the T(6) insulin hexamer. An unusual feature of each molecule in the dimer was the presence of two independent conformations at the B-chain C-terminus (residues B20-B25). Both conformations were different from that of native insulin, involving a 3.5 A displacement of the B20-B23 beta-turn and a repositioning of residue PheB25 such that it made close van der Waals contact with the main body of the molecule, appearing to stabilize the B-chain C-terminus.

Synthesis and characterization of monomeric and dimeric manganese(II and zinc(II complexes of pyridine-2-carbaldoxime

Directory of Open Access Journals (Sweden)

Jørgen Glerup

2000-12-01

Full Text Available The syntheses and characterization of two complexes of manganese(II and one complex of zinc(II with the ligand pyridine-2-carbaldoxime, C6H6N2O, are described. The monomeric manganese(II complex cis-[Mn(C6H6N2O 2Cl2] (1 crystallizes in the orthorhombic space group Pbcn with 4 formula units in a cell of dimensions a = 12.479(3 Å, b = 10.348(2 Å, and c = 11. 974(2 Å. The structure has been refined to a final value of the conventional R-factor of 0.0330 based on 1513 observed independent reflections. The analogous zinc(II complex, cis-[Zn(C6H6N2O2Cl2] (2 also crystallizes in the orthorhombic space group Pbcn with 4 formula units in a cell of dimensions a = 12.215(2 Å, b = 10.383(2 Å, and c = 12. 016(2 Å. The structure has been refined to a final value of the conventional R-factor of 0.0377 based on 1117 observed independent reflections. The two complexes are isostructural, with the central metal atom lying on a crystallographic 2-fold axis. Both complexes are approximately octahedral, the coordination being provided by two trans pyridine nitrogen atoms and two cis amine nitrogen atoms from the oxime ligands, and by two cis chlorides. The dimeric manganese(II complex [(C6H6N2O(CH3OHClMnCl2MnCl(CH3OH(C6H6N2O] (3 crystallizes in the monoclinic space group P21/n with 2 formula units in a cell of dimensions a = 7.895(2 Å, b = 11.196(3 Å, and c = 12. 544(2 Å, and b = 98.39(2o. The structure has been refined to a final value of the conventional R-factor of 0.0312 based on 1568 observed independent reflections. There is a crystallographic inversion center in the middle of the dimer relating one manganese center to the other. The geometry at each manganese(II center is again roughly octahedral, coordination being provided by two nitrogen atoms from the oxime ligand, a terminal chloride ion trans to the amine nitrogen, the oxygen atom of the coordinated methanol molecule, and two bridging chlorides that link the two halves of the dimer. The Mn

Principles of DNA architectonics: design of DNA-based nanoobjects

International Nuclear Information System (INIS)

Vinogradova, O A; Pyshnyi, D V

2012-01-01

The methods of preparation of monomeric DNA blocks that serve as key building units for the construction of complex DNA objects are described. Examples are given of the formation of DNA blocks based on native and modified oligonucleotide components using hydrogen bonding and nucleic acid-specific types of bonding and also some affinity interactions with RNA, proteins, ligands. The static discrete and periodic two- and three-dimensional DNA objects reported to date are described systematically. Methods used to prove the structures of DNA objects and the prospects for practical application of nanostructures based on DNA and its analogues in biology, medicine and biophysics are considered. The bibliography includes 195 references.

Turbulent transport in reversed field pinches

International Nuclear Information System (INIS)

Christiansen, J.P.; Roberts, K.V.

1976-01-01

MHD stability of the Reversed Field Pinch (RFP) relies on reversal of the toroidal field component in the outer plasma region. Interest in this configuration comes from its potential economic advantages as a thermonuclear reactor, since compared to a Tokamak the RFP supports a higher value of β, the ratio between plasma and total magnetic pressure. Results of computations on the time-evolution of the RFP using a 1D MHD model are reported. (orig./GG) [de

RFP Patterns and Techniques for Successful Agile Contracting

Science.gov (United States)

2016-11-01

5.5 Risk Assessment 27 5.6 Development Methodologies 27 5.6.1 The Waterfall Model 28 5.6.2 The Agile Model 29 5.7 Mixing Waterfall and Agile...familiarity with sequential, waterfall -based software development models . A brief refresher is provided in Section 5.6.1. 4 See AcqNotes...Environment In a traditional waterfall model , technical reviews are used as control gates to move from one se- quential phase to the next (e.g

Compact reversed-field pinch reactors (CRFPR)

International Nuclear Information System (INIS)

Krakowski, R.A.; Hagenson, R.L.; Schnurr, N.M.; Copenhaver, C.; Bathke, C.G.; Miller, R.L.; Embrechts, M.J.

1986-01-01

The unique confinement properties of the poloidal-field-dominated Reversed-Field Pinch (RFP) are exploited to examine physics and technical issues related to a compact high-power-density fusion reactor. This resistive-coil, steady-state, toroidal device would use a dual-media (i.e., two separate coolants) power cycle that would be driven by a fusion power core (FPC, i.e., plasma chamber, first wall, blanket, shield, and coils) having a power density and mass approaching pressurized-water-fission reactor values. A 1000-MWe(net) base case is selected from a comprehensive trade-off study to examine technological issues related to operating a high-power-density FPC. A general rationale outlining the need for improved fusion concepts is given, followed by a description of the RFP principle, a detailed systems and trade-off analysis, and a conceptual FPC design for the ∝ 20-MW/m 2 (neutrons) compact RFP reactor, CRFPR(20). Key FPC components are quantified, and full power-balance, thermal, and mechanical FPC integrations are given. (orig.)

Develop Improved Materials to Support the Hydrogen Economy

Energy Technology Data Exchange (ETDEWEB)

Dr. Michael C. Martin

2012-07-18

The Edison Materials Technology Center (EMTEC) solicited and funded hydrogen infrastructure related projects that have a near term potential for commercialization. The subject technology of each project is related to the US Department of Energy hydrogen economy goals as outlined in the multi-year plan titled, 'Hydrogen, Fuel Cells and Infrastructure Technologies Program Multi-Year Research, Development and Demonstration Plan.' Preference was given to cross cutting materials development projects that might lead to the establishment of manufacturing capability and job creation. The Edison Materials Technology Center (EMTEC) used the US Department of Energy hydrogen economy goals to find and fund projects with near term commercialization potential. An RFP process aligned with this plan required performance based objectives with go/no-go technology based milestones. Protocols established for this program consisted of a RFP solicitation process, white papers and proposals with peer technology and commercialization review (including DoE), EMTEC project negotiation and definition and DoE cost share approval. Our RFP approach specified proposals/projects for hydrogen production, hydrogen storage or hydrogen infrastructure processing which may include sensor, separator, compression, maintenance, or delivery technologies. EMTEC was especially alert for projects in the appropriate subject area that have cross cutting materials technology with near term manufacturing and commercialization opportunities.

Validación de un método de cromatografía líquida para la determinación de rifampicina en plasma humano

Directory of Open Access Journals (Sweden)

Luis Moreno-Exebio

Full Text Available Objetivos: Validar un método de cromatografía líquida de alta resolución (HPLC para la determinación de rifampicina (RFP en plasma humano. Materiales y métodos. Se desarrolló un método HPLC para la determinación de RFP en plasma. La separación fue realizada por cromatografía de fase reversa con una columna C18 y una fase móvil compuesta por una mezcla de acetonitrilo y solución amortiguadora de fosfato de potasio monobásico 0,05 M (38:62 v/v a 335 nm. En el cual se empleó como estándar interno rifampicina quinona (RFP-QN. Resultados. Los tiempos de retención de RFP y RFP-QN fueron 7,81 y 12,26 minutos, respectivamente. El ensayo fue lineal de 0,5 a 250 ug/mL Los parámetros evaluados de precisión, exactitud, selectividad, linealidad, recuperación cumplieron con lo establecido en las guías internacionales de validación de métodos bioanalíticos. Conclusiones. El método HPLC desarrollado es simple, específico, sensible, selectivo y lineal para un amplio rango de concentraciones de RFP en plasma

A construct with fluorescent indicators for conditional expression of miRNA

Directory of Open Access Journals (Sweden)

Xia Xugang

2008-10-01

Full Text Available Abstract Background Transgenic RNAi holds promise as a simple, low-cost, and fast method for reverse genetics in mammals. It may be particularly useful for producing animal models for hypomorphic gene function. Inducible RNAi that permits spatially and temporally controllable gene silencing in vivo will enhance the power of transgenic RNAi approach. Furthermore, because microRNA (miRNA targeting specific genes can be expressed simultaneously with protein coding genes, incorporation of fluorescent marker proteins can simplify the screening and analysis of transgenic RNAi animals. Results We sought to optimally express a miRNA simultaneously with a fluorescent marker. We compared two construct designs. One expressed a red fluorescent protein (RFP and a miRNA placed in its 3' untranslated region (UTR. The other expressed the same RFP and miRNA, but the precursor miRNA (pre-miRNA coding sequence was placed in an intron that was inserted into the 3'-UTR. We found that the two constructs expressed comparable levels of miRNA. However, the intron-containing construct expressed a significantly higher level of RFP than the intron-less construct. Further experiments indicate that the 3'-UTR intron enhances RFP expression by its intrinsic gene-expression-enhancing activity and by eliminating the inhibitory effect of the pre-miRNA on the expression of RFP. Based on these findings, we incorporated the intron-embedded pre-miRNA design into a conditional expression construct that employed the Cre-loxP system. This construct initially expressed EGFP gene, which was flanked by loxP sites. After exposure to Cre recombinase, the transgene stopped EGFP expression and began expression of RFP and a miRNA, which silenced the expression of specific cellular genes. Conclusion We have designed and tested a conditional miRNA-expression construct and showed that this construct expresses both the marker genes strongly and can silence the target gene efficiently upon Cre

Improving brightness and photostability of green and red fluorescent proteins for live cell imaging and FRET reporting.

Science.gov (United States)

Bajar, Bryce T; Wang, Emily S; Lam, Amy J; Kim, Bongjae B; Jacobs, Conor L; Howe, Elizabeth S; Davidson, Michael W; Lin, Michael Z; Chu, Jun

2016-02-16

Many genetically encoded biosensors use Förster resonance energy transfer (FRET) to dynamically report biomolecular activities. While pairs of cyan and yellow fluorescent proteins (FPs) are most commonly used as FRET partner fluorophores, respectively, green and red FPs offer distinct advantages for FRET, such as greater spectral separation, less phototoxicity, and lower autofluorescence. We previously developed the green-red FRET pair Clover and mRuby2, which improves responsiveness in intramolecular FRET reporters with different designs. Here we report the engineering of brighter and more photostable variants, mClover3 and mRuby3. mClover3 improves photostability by 60% and mRuby3 by 200% over the previous generation of fluorophores. Notably, mRuby3 is also 35% brighter than mRuby2, making it both the brightest and most photostable monomeric red FP yet characterized. Furthermore, we developed a standardized methodology for assessing FP performance in mammalian cells as stand-alone markers and as FRET partners. We found that mClover3 or mRuby3 expression in mammalian cells provides the highest fluorescence signals of all jellyfish GFP or coral RFP derivatives, respectively. Finally, using mClover3 and mRuby3, we engineered an improved version of the CaMKIIα reporter Camuiα with a larger response amplitude.

Use of the quartz crystal microbalance to determine the monomeric friction coefficient of polyimides

Science.gov (United States)

Bechtold, Mary M.

1995-01-01

When a thin film of polymer is coated on to a quartz crystal microbalance (QCM), the QCM can be used to detect the rate of increase in weight of the polymer film as the volatile penetrant diffuses into the polymer. From this rate information the diffusion coefficient of the penetrant into the polymer can be computed. Calculations requiring this diffusion coefficient lead to values which approximate the monomeric friction coefficient of the polymer. This project has been concerned with the trial of crystal oscillating circuits suitable for driving polymer coated crystals in an atmosphere of penetrant. For these studies done at room temperature, natural rubber was used as an easily applied polymer that is readily penetrated by toluene vapors, qualities anticipated with polyimides when they are tested at T(g) in the presence of toluene. Three quartz crystal oscillator circuits were tested. The simplest circuit used +/- 5 volt dc and had a transistor to transistor logic (TTL) inverter chip that provides a 180 deg phase shift via a feed back loop. This oscillator circuit was stable but would not drive the crystal when the crystal was coated with polymer and subjected to toluene vapors. Removal of a variable resistor from this circuit increased stability but did not otherwise increase performance. Another driver circuit tested contained a two stage differential input, differential output, wide band video amplifier and also contain a feed back loop. The circuit voltage could not be varied and operated at +/- 5 volts dc; this circuit was also stable but failed to oscillate the polymer coated crystal in an atmosphere saturated with toluene vapors. The third oscillator circuit was of similar construction and relied on the same video amplifier but allowed operation with variable voltage. This circuit would drive the crystal when the crystal was submerged in liquid toluene and when the crystal was coated with polymer and immersed in toluene vapors. The frequency readings

Resonance Raman assignment and evidence for noncoupling of individual 2- and 4-vinyl vibrational modes in a monomeric cyanomethemoglobin

International Nuclear Information System (INIS)

Gersonde, K.; Yu, N.T.; Lin, S.H.; Smith, K.M.; Parish, D.W.

1989-01-01

We have investigated the resonance Raman spectra of monomeric insect cyanomethemoglobins (CTT III and CTT IV) reconstituted with (1) protohemes IX selectively deuterated at the 4-vinyl as well as the 2,4-divinyls, (2) monovinyl-truncated hemes such as pemptoheme (2-hydrogen, 4-vinyl) and isopemptoheme (2-vinyl, 4-hydrogen), (3) symmetric hemes such as protoheme III (with 2- and 3-vinyls) and protoheme XIII (with 1- and 4-vinyls), and (4) hemes without 2- and 4-vinyls such as mesoheme IX, deuteroheme IX, 2,4-dimethyldeuteroheme IX, and 2,4-dibromodeuteroheme IX. Evidence is presented that the highly localized vinyl C = C stretching vibrations at the 2- and 4-positions of the heme in these cyanomet CTT hemoglobins are noncoupled and inequivalent; i.e., the 1631- and 1624-cm-1 lines have been assigned to 2-vinyl and 4-vinyl, respectively. The elimination of the 2-vinyl (in pemptoheme) or the 4-vinyl (in isopemptoheme) does not affect the C = C stretching frequency of the remaining vinyl. Furthermore, two low-frequency vinyl bending modes at 412 and 591 cm-1 exhibit greatly different resonance Raman intensities between 2-vinyl and 4-vinyl. The observed intensity at 412 cm-1 is primarily derived from 4-vinyl, whereas the 591-cm-1 line results exclusively from the 2-vinyl. Again, there is no significant coupling between 2-vinyl and 4-vinyl for these two bending modes

Heteromeric p97/p97R155C complexes induce dominant negative changes in wild-type and autophagy 9-deficient Dictyostelium strains.

Directory of Open Access Journals (Sweden)

Khalid Arhzaouy

Full Text Available Heterozygous mutations in the human VCP (p97 gene cause autosomal-dominant IBMPFD (inclusion body myopathy with early onset Paget's disease of bone and frontotemporal dementia, ALS14 (amyotrophic lateral sclerosis with or without frontotemporal dementia and HSP (hereditary spastic paraplegia. Most prevalent is the R155C point mutation. We studied the function of p97 in the social amoeba Dictyostelium discoideum and have generated strains that ectopically express wild-type (p97 or mutant p97 (p97(R155C fused to RFP in AX2 wild-type and autophagy 9 knock-out (ATG9(KO cells. Native gel electrophoresis showed that both p97 and p97(R155C assemble into hexamers. Co-immunoprecipitation studies revealed that endogenous p97 and p97(R155C-RFP form heteromers. The mutant strains displayed changes in cell growth, phototaxis, development, proteasomal activity, ubiquitinylated proteins, and ATG8(LC3 indicating mis-regulation of multiple essential cellular processes. Additionally, immunofluorescence analysis revealed an increase of protein aggregates in ATG9(KO/p97(R155C-RFP and ATG9(KO cells. They were positive for ubiquitin in both strains, however, solely immunoreactive for p97 in the ATG9(KO mutant. A major finding is that the expression of p97(R155C-RFP in the ATG9(KO strain partially or fully rescued the pleiotropic phenotype. We also observed dose-dependent effects of p97 on several cellular processes. Based on findings in the single versus the double mutants we propose a novel mode of p97 interaction with the core autophagy protein ATG9 which is based on mutual inhibition.

Color-Coded Imaging of Syngeneic Orthotopic Malignant Lymphoma Interacting with Host Stromal Cells During Metastasis.

Science.gov (United States)

Matsumoto, Takuro; Suetsugu, Atsushi; Hasegawa, Kosuke; Nakamura, Miki; Aoki, Hitomi; Kunisada, Takahiro; Tsurumi, Hisashi; Shimizu, Masahito; Hoffman, Robert M

2016-04-01

The EL4 cell line was previously derived from a lymphoma induced in a C57/BL6 mouse by 9,10-dimethyl-1,2-benzanthracene. In a previous study, EL4 lymphoma cells expressing red fluorescent protein (EL4-RFP) were established and injected into the tail vein of C57/BL6 green fluorescent protein (GFP) transgenic mice. Metastasis was observed at multiple sites which were also enriched with host GFP-expressing stromal cells. In the present study, our aim was to establish an orthotopic model of EL4-RFP. In the present study, EL4-RFP lymphoma cells were injected in the spleen of C57/BL6 GFP transgenic mice as an orthotopic model of lymphoma. Resultant primary tumor and metastases were imaged with the Olympus FV1000 scanning laser confocal microscope. EL4-RFP metastasis was observed 21 days later. EL4-RFP tumors in the spleen (primary injection site), liver, supra-mediastinum lymph nodes, abdominal lymph nodes, bone marrow, and lung were visualized by color-coded imaging. EL4-RFP metastases in the liver, lymph nodes, and bone marrow in C57/BL6 GFP mice were rich in GFP stromal cells such as macrophages, fibroblasts, dendritic cells, and normal lymphocytes derived from the host animal. Small tumors were observed in the spleen, which were rich in host stromal cells. In the lung, no mass formation of lymphoma cells occurred, but lymphoma cells circulated in lung peripheral blood vessels. Phagocytosis of EL4-RFP lymphoma cells by macrophages, as well as dendritic cells and fibroblasts, were observed in culture. Color-coded imaging of the lymphoma microenvironment suggests an important role of stromal cells in lymphoma progression and metastasis. Copyright© 2016 International Institute of Anticancer Research (Dr. John G. Delinassios), All rights reserved.

An Autophagic Flux Probe that Releases an Internal Control.

Science.gov (United States)

Kaizuka, Takeshi; Morishita, Hideaki; Hama, Yutaro; Tsukamoto, Satoshi; Matsui, Takahide; Toyota, Yuichiro; Kodama, Akihiko; Ishihara, Tomoaki; Mizushima, Tohru; Mizushima, Noboru

2016-11-17

Macroautophagy is an intracellular degradation system that utilizes the autophagosome to deliver cytoplasmic components to the lysosome. Measuring autophagic activity is critically important but remains complicated and challenging. Here, we have developed GFP-LC3-RFP-LC3ΔG, a fluorescent probe to evaluate autophagic flux. This probe is cleaved by endogenous ATG4 proteases into equimolar amounts of GFP-LC3 and RFP-LC3ΔG. GFP-LC3 is degraded by autophagy, while RFP-LC3ΔG remains in the cytosol, serving as an internal control. Thus, autophagic flux can be estimated by calculating the GFP/RFP signal ratio. Using this probe, we re-evaluated previously reported autophagy-modulating compounds, performed a high-throughput screen of an approved drug library, and identified autophagy modulators. Furthermore, we succeeded in measuring both induced and basal autophagic flux in embryos and tissues of zebrafish and mice. The GFP-LC3-RFP-LC3ΔG probe is a simple and quantitative method to evaluate autophagic flux in cultured cells and whole organisms. Copyright © 2016 Elsevier Inc. All rights reserved.

Plasma behaviour in large reversed-field pinches and reactors

International Nuclear Information System (INIS)

Christiansen, J.P.; Bodin, H.A.B.; Carolan, P.G.; Johnston, J.W.; Newton, A.A.; Roberts, K.V.; Robinson, D.C.; Watts, M.R.C.; Piotrowicz, V.A.

1981-01-01

Recent analytic and numerical results on large reversed-field-pinch (RFP) systems and RFP reactors are presented. Predictions are made of the plasma behaviour in Eta Beta II, HBTXIA (under construction) and RFX (planned). The setting-up phase of an RFP is studied by using turbulence theory in transport equilibrium calculations, and estimates are made of the volt-seconds consumption for four different modes of field control. A prescription is given for a dynamo producing self-reversal which yields finite-β configurations. Residual instabilities of these equilibria may be resistive pressure-driven g-modes, and a new study of these modes that includes parallel viscosity indicates stability for anti β approximately 10%. The sustainment phase of the RFP is examined with tokamak scaling laws assumed for the energy confinement time. Temperatures in excess of 1keV are predicted for currents of 2MA in RFX. An operating cycle for a pulsed RFP reactor including gas puffing to reach ignition is proposed following a study of the energy replacement time for an Ohmically heated plasma. The scaling of the reactor parameters with minor radius is also investigated. (author)

Characterization of uranium in surface-waters collected at the Rocky Flats Facility

International Nuclear Information System (INIS)

Efurd, D.W.; Rokop, D.J.; Aguilar, R.D.; Roensch, F.R.; Perrin, R.E.; Banar, J.C.

1994-01-01

The Rocky Flats Plant (RFP) is a Department of Energy (DOE) facility where plutonium and uranium components were manufactured for nuclear weapons. During plant operations radioactivity was inadvertently released into the environment. This study was initiated to characterize the uranium present in surface-waters at RFP. Three drainage basins and natural ephemeral streams transverse RFP. The Woman Creek drainage basin traverses and drains the southern portion of the site. The Rock Creek drainage basin drains the northwestern portion of the plant complex. The Walnut Creek drainage basin traverses the western, northern, and northeastern portions of the RFP site. Dams, detention ponds, diversion structures, and ditches have been constructed at RFP to control the release of plant discharges and surface (storm water) runoff. The ponds located downstream of the plant complex on North Walnut Creek are designated A-1 through A-4. Ponds on South Walnut Creek are designated B-1 through B-5. The ponds in the Woman Creek drainage basin are designated C-1 and C-2. Water samples were collected from each pond and the uranium was characterized by TIMS measurement techniques

Selecting practice management information systems.

Science.gov (United States)

Worley, R; Ciotti, V

1997-01-01

Despite enormous advances in information systems, the process by which most medical practices select them has remained virtually unchanged for decades: the request for proposal (RFP). Unfortunately, vendors have learned ways to minimize the value of RFP checklists to where purchasers now learn little about the system functionality. The authors describe a selection methodology that replaces the RFP with scored demos, reviews of vendor user manuals and mathematically structured reference checking. In a recent selection process at a major medical center, these techniques yielded greater user buy-in and favorable contract terms as well.

MHD computation of feedback of resistive-shell instabilities in the reversed field pinch

International Nuclear Information System (INIS)

Zita, E.J.; Prager, S.C.

1992-05-01

MHD computation demonstrates that feedback can sustain reversal and reduce loop voltage in resistive-shell reversed field pinch (RFP) plasmas. Edge feedback on ∼2R/a tearing modes resonant near axis is found to restore plasma parameters to nearly their levels with a close-fitting conducting shell. When original dynamo modes are stabilized, neighboring tearing modes grow to maintain the RFP dynamo more efficiently. This suggests that experimentally observed limits on RFP pulselengths to the order of the shell time can be overcome by applying feedback to a few helical modes

Hue-shifted monomeric variants of Clavularia cyan fluorescent protein: identification of the molecular determinants of color and applications in fluorescence imaging

Directory of Open Access Journals (Sweden)

Davidson Michael W

2008-03-01

Full Text Available Abstract Background In the 15 years that have passed since the cloning of Aequorea victoria green fluorescent protein (avGFP, the expanding set of fluorescent protein (FP variants has become entrenched as an indispensable toolkit for cell biology research. One of the latest additions to the toolkit is monomeric teal FP (mTFP1, a bright and photostable FP derived from Clavularia cyan FP. To gain insight into the molecular basis for the blue-shifted fluorescence emission we undertook a mutagenesis-based study of residues in the immediate environment of the chromophore. We also employed site-directed and random mutagenesis in combination with library screening to create new hues of mTFP1-derived variants with wavelength-shifted excitation and emission spectra. Results Our results demonstrate that the protein-chromophore interactions responsible for blue-shifting the absorbance and emission maxima of mTFP1 operate independently of the chromophore structure. This conclusion is supported by the observation that the Tyr67Trp and Tyr67His mutants of mTFP1 retain a blue-shifted fluorescence emission relative to their avGFP counterparts (that is, Tyr66Trp and Tyr66His. Based on previous work with close homologs, His197 and His163 are likely to be the residues with the greatest contribution towards blue-shifting the fluorescence emission. Indeed we have identified the substitutions His163Met and Thr73Ala that abolish or disrupt the interactions of these residues with the chromophore. The mTFP1-Thr73Ala/His163Met double mutant has an emission peak that is 23 nm red-shifted from that of mTFP1 itself. Directed evolution of this double mutant resulted in the development of mWasabi, a new green fluorescing protein that offers certain advantages over enhanced avGFP (EGFP. To assess the usefulness of mTFP1 and mWasabi in live cell imaging applications, we constructed and imaged more than 20 different fusion proteins. Conclusion Based on the results of our

Compact reversed-field pinch reactors (CRFPR): sensitivity study and design-point determination

International Nuclear Information System (INIS)

Hagenson, R.L.; Krakowski, R.A.

1982-07-01

If the costing assumptions upon which the positive assessment of conventional large superconducting fusion reactors are based proves overly optimistic, approaches that promise considerably increased system power density and reduced mass utilization will be required. These more compact reactor embodiments generally must operate with reduced shield thickness and resistive magnets. Because of the unique, magnetic topology associated with the Reversed-Field Pinch (RFP), the compact reactor embodiment for this approach is particularly attractive from the viewpoint of low-field resistive coils operating with Ohmic losses that can be made small relative to the fusion power. A comprehensive system model is developed and described for a steady-state, compact RFP reactor (CRFPR). This model is used to select a unique cost-optimized design point that will be used for a conceptual engineering design. The cost-optimized CRFPR design presented herein would operate with system power densities and mass utilizations that are comparable to fission power plants and are an order of magnitude more favorable than the conventional approaches to magnetic fusion power. The sensitivity of the base-case design point to changes in plasma transport, profiles, beta, blanket thickness, normal vs superconducting coils, and fuel cycle (DT vs DD) is examined. The RFP approach is found to yield a point design for a high-power-density reactor that is surprisingly resilient to changes in key, but relatively unknown, physics and systems parameters

Interaction of chemokine receptor CXCR4 in monomeric and dimeric state with its endogenous ligand CXCL12: coarse-grained simulations identify differences.

Science.gov (United States)

Cutolo, Pasquale; Basdevant, Nathalie; Bernadat, Guillaume; Bachelerie, Françoise; Ha-Duong, Tâp

2017-02-01

Despite the recent resolutions of the crystal structure of the chemokine receptor CXCR4 in complex with small antagonists or viral chemokine, a description at the molecular level of the interactions between the full-length CXCR4 and its endogenous ligand, the chemokine CXCL12, in relationship with the receptor recognition and activation, is not yet completely elucidated. Moreover, since CXCR4 is able to form dimers, the question of whether the CXCR4-CXCL12 complex has a 1:1 or 2:1 preferential stoichiometry is still an open question. We present here results of coarse-grained protein-protein docking and molecular dynamics simulations of CXCL12 in association with CXCR4 in monomeric and dimeric states. Our proposed models for the 1:1 and 2:1 CXCR4-CXCL12 quaternary structures are consistent with recognition and activation motifs of both partners provided by the available site-directed mutagenesis data. Notably, we observed that in the 2:1 complex, the chemokine N-terminus makes more steady contacts with the receptor residues critical for binding and activation than in the 1:1 structure, suggesting that the 2:1 stoichiometry would favor the receptor signaling activity with respect to the 1:1 association.

Promoter, transgene, and cell line effects in the transfection of mammalian cells using PDMAEMA-based nano-stars

Directory of Open Access Journals (Sweden)

Alexander Raup

2016-09-01

Full Text Available Non-viral transfection protocols are typically optimized using standard cells and reporter proteins, potentially underestimating cellular or transgene effects. Here such effects were studied for two human (Jurkat, HEK-293 and two rodent (CHO-K1, L929 cell lines and three fluorescent reporter proteins. Expression of the enhanced green fluorescent protein (EGFP was studied under the control of the human elongation factor 1 alpha promoter and three viral promoters (SV40, SV40/enhancer, CMV, that of ZsYellow1 (yellow fluorescence and mCherry (red fluorescence for the CMV promoter. Results varied with the cell line, in particular for the Jurkat cells. Pair-wise co-transfection of the CMV controlled transgenes resulted in a significant fraction of monochromatic cells (EGFP for EGFP/YFP and EGFP/RFP co-transfections, YFP in case of YFP/RFP co-transfections. Only Jurkat cells were almost incapable of expressing YFP. Dilution of the plasmid DNA with a non-expressed plasmid showed cell line dependent effects on transfection efficiency and/or expression levels.

Equilibrium measurements on the REPUTE-1 RFP plasma

International Nuclear Information System (INIS)

Ji, H.; Toyama, H.; Shinohara, S.; Fujisawa, A.; Yamagishi, K.; Shimazu, Y.; Ejiri, A.; Shimoji, K.; Miyamoto, K.

1989-01-01

Global plasma equilibrium measurements by external magnetic probes are widely introduced on the toroidal plasmas, i.e. tokamaks or RFPs, because of their simplicity and convenience. The measurement principle is based on Shafranov's toroidal equilibrium theory, which gives simple relations between the global equilibrium quantity and the external fields. These relations are valid in the either case of existence or absence of ideal shell just out the plasma column, however, not valid in the case of the thin (or resistive) shell, whose skin time τ s has the same order of the current rise time τ r . A method introduced by Swain et al. is effective in this case, in which the plasma current I p is replaced by 6 filament currents. However, by this method it is dificult to include the effect of iron core and computation requires a lot (beyond 14) of the measurement of the fields or flux loop. In this paper we introduce a simple method which is based on fitting measured fields to the vacuum approximate solution of Grad-Shafranov equation. The computation requires only a few measurements (≥6) of the fields. REPUTE-1 device is characterized by a thin shell of 5 mm thickness whose skin time τ s for the penetration of the vertical field is 1 ms compared with τ r of 0.5 ms. The optimum discharges whose duration τ d are about 3 times of τ s have been obtained. In spite of various efforts including vertical-ohmic coils series connection experiments, toroidal ripple reduction experiments and port bypass plate installation experiments, until now τ d is still limited by 3.2 ms. We should think that the equilibrium of plasma is lost due to an unfavorable vertical field. In this paper we present the measurements of the time evolution of the plasma position from the flat-top phase to the termination phase, at that time the plasma begins to lose its equilibrium. The liner has a major radius R L of 82 cm and a minor radius a L of 22 cm. (author) 6 refs., 4 figs

78 FR 55029 - Approval and Promulgation of Implementation; Texas; Houston: Reasonable Further Progress Plan...

Science.gov (United States)

2013-09-09

...): Transportation Control Measures and Transportation Control Strategies To Offset Growth in Emissions Due to Growth... progress (RFP) plan and contingency measures, the vehicle miles traveled (VMT) offset analysis, and... and Emission Reductions for RFP E. Contingency Measures F. Vehicle Miles Traveled Offset Analysis G...

77 FR 5032 - National Cancer Institute; Notice of Meeting

Science.gov (United States)

2012-02-01

... Initiatives; RFA and RFP Concept Reviews; and Scientific Presentations. Place: National Institutes of Health... Group(s); and Budget Presentations; Reports of Special Initiatives; RFA and RFP Concept Reviews; and Scientific Presentations. Place: National Institutes of Health, Building 31, 31 Center Drive, 6th Floor, Conf...

Environmental Survey preliminary report, Rocky Flats Plant, Golden, Colorado

International Nuclear Information System (INIS)

1987-06-01

This report presents the preliminary findings of the Environmental Survey of the United States Department of Energy (DOE), Rocky Flats Plant (RFP), conducted August 11 through 22, 1986. The Survey is being conducted by an multidisciplinary team of environmental specialists, led and managed by the Office of Environment, Safety and Health's Office of Environmental Audit. Individual team members are outside experts supplied by a private contractor. The objective of the Survey is to identify environmental problems and areas of environmental risk associated with the RFP. The Survey covers all environmental media and all areas of environmental regulations. It is being performed in accordance with the DOE Environmental Survey Manual. The on-site phase of the Survey involves the review of existing site environmental data observations of the operations carried on at RFP, and interviews with site personnel. The Survey team developed a Sampling and Analysis Plan to assist in further assessing certain environmental problems identified during its on-site activates. The Sampling and Analysis Plan is being executed by DOE's Oak Ridge National Laboratory. When completed, the results will be incorporated into the RFP Environmental Survey Interim Report. The Interim Report will reflect the final determinations of the RFP Survey. 75 refs., 24 figs., 33 tabs

Environmental Survey preliminary report, Rocky Flats Plant, Golden, Colorado

Energy Technology Data Exchange (ETDEWEB)

1987-06-01

This report presents the preliminary findings of the Environmental Survey of the United States Department of Energy (DOE), Rocky Flats Plant (RFP), conducted August 11 through 22, 1986. The Survey is being conducted by an multidisciplinary team of environmental specialists, led and managed by the Office of Environment, Safety and Health's Office of Environmental Audit. Individual team members are outside experts supplied by a private contractor. The objective of the Survey is to identify environmental problems and areas of environmental risk associated with the RFP. The Survey covers all environmental media and all areas of environmental regulations. It is being performed in accordance with the DOE Environmental Survey Manual. The on-site phase of the Survey involves the review of existing site environmental data observations of the operations carried on at RFP, and interviews with site personnel. The Survey team developed a Sampling and Analysis Plan to assist in further assessing certain environmental problems identified during its on-site activates. The Sampling and Analysis Plan is being executed by DOE's Oak Ridge National Laboratory. When completed, the results will be incorporated into the RFP Environmental Survey Interim Report. The Interim Report will reflect the final determinations of the RFP Survey. 75 refs., 24 figs., 33 tabs.

Reversed-field-pinch and ultra-low-q discharges in REPUTE-2

International Nuclear Information System (INIS)

Inoue, N.; Yoshida, Z.; Kamada, Y.; Saito, M.; Miyamoto, K.

1987-01-01

Ultra-low q (ULQ) and very-low q (VLQ) discharge experiments have been done using the REPUTE-1 RFP. It was found that in these q regime, the plasma density and beta are fairly high, and the confinement property is less sensitive to the error field compared to the RFP. However, since the temperature of the REPUTE-1 discharge is limited in low value because of the small plasma current due to the small toroidal field, its magnetic Reynolds number is too small to simulate the reactor plasma behavior. The radiation barrier has not been overcome yet, and consequently the energy confinement time is very short. In order to improve these aspects of the REPUTE-1 experiment, the REPUTE-2 is designed to produce higher toroidal field of 2T. The toroidal field increases slowly to the final value as in the case of the ramp-up mode of the RFP operation. The first stage of the REPUTE-2 project will be devoted to study the confinement physics of RFP, ULQ, and VLQ. In the second stage, innovation of these configurations, such as resistive shell RFP, neutral beam current drive, and higher current density, is planned. 8 refs., 1 fig., 2 tabs

Tolerance of a knotted near infrared fluorescent protein to random circular permutation

Science.gov (United States)

Pandey, Naresh; Kuypers, Brianna E.; Nassif, Barbara; Thomas, Emily E.; Alnahhas, Razan N.; Segatori, Laura; Silberg, Jonathan J.

2016-01-01

Bacteriophytochrome photoreceptors (BphP) are knotted proteins that have been developed as near-infrared fluorescent protein (iRFP) reporters of gene expression. To explore how rearrangements in the peptides that interlace into the knot within the BphP photosensory core affect folding, we subjected iRFP to random circular permutation using an improved transposase mutagenesis strategy and screened for variants that fluoresce. We identified twenty seven circularly permuted iRFP that display biliverdin-dependent fluorescence in Escherichia coli. The variants with the brightest whole cell fluorescence initiated translation at residues near the domain linker and knot tails, although fluorescent variants were discovered that initiated translation within the PAS and GAF domains. Circularly permuted iRFP retained sufficient cofactor affinity to fluoresce in tissue culture without the addition of biliverdin, and one variant displayed enhanced fluorescence when expressed in bacteria and tissue culture. This variant displayed a similar quantum yield as iRFP, but exhibited increased resistance to chemical denaturation, suggesting that the observed signal increase arose from more efficient protein maturation. These results show how the contact order of a knotted BphP can be altered without disrupting chromophore binding and fluorescence, an important step towards the creation of near-infrared biosensors with expanded chemical-sensing functions for in vivo imaging. PMID:27304983

20 CFR 411.310 - How does an entity other than a State VR agency apply to be an EN and who will determine whether...

Science.gov (United States)

2010-04-01

....310 Employees' Benefits SOCIAL SECURITY ADMINISTRATION THE TICKET TO WORK AND SELF-SUFFICIENCY PROGRAM... responding to our Request for Proposal (RFP), which we published in the Commerce Business Daily and which is available online through the Federal government's electronic posting system (http://www.eps.gov). This RFP...

Monitoring thioredoxin redox with a genetically encoded red fluorescent biosensor.

Science.gov (United States)

Fan, Yichong; Makar, Merna; Wang, Michael X; Ai, Hui-Wang

2017-09-01

Thioredoxin (Trx) is one of the two major thiol antioxidants, playing essential roles in redox homeostasis and signaling. Despite its importance, there is a lack of methods for monitoring Trx redox dynamics in live cells, hindering a better understanding of physiological and pathological roles of the Trx redox system. In this work, we developed the first genetically encoded fluorescent biosensor for Trx redox by engineering a redox relay between the active-site cysteines of human Trx1 and rxRFP1, a redox-sensitive red fluorescent protein. We used the resultant biosensor-TrxRFP1-to selectively monitor perturbations of Trx redox in various mammalian cell lines. We subcellularly localized TrxRFP1 to image compartmentalized Trx redox changes. We further combined TrxRFP1 with a green fluorescent Grx1-roGFP2 biosensor to simultaneously monitor Trx and glutathione redox dynamics in live cells in response to chemical and physiologically relevant stimuli.

In vitro culture and characterization of human lung cancer circulating tumor cells isolated by size exclusion from an orthotopic nude-mouse model expressing fluorescent protein.

Science.gov (United States)

Kolostova, Katarina; Zhang, Yong; Hoffman, Robert M; Bobek, Vladimir

2014-09-01

In the present study, we demonstrate an animal model and recently introduced size-based exclusion method for circulating tumor cells (CTCs) isolation. The methodology enables subsequent in vitro CTC-culture and characterization. Human lung cancer cell line H460, expressing red fluorescent protein (H460-RFP), was orthotopically implanted in nude mice. CTCs were isolated by a size-based filtration method and successfully cultured in vitro on the separating membrane (MetaCell®), analyzed by means of time-lapse imaging. The cultured CTCs were heterogeneous in size and morphology even though they originated from a single tumor. The outer CTC-membranes were blebbing in general. Abnormal mitosis resulting in three daughter cells was frequently observed. The expression of RFP ensured that the CTCs originated from lung tumor. These readily isolatable, identifiable and cultivable CTCs can be used to characterize individual patient cancers and for screening of more effective treatment.

Production of Monomeric Aromatic Compounds from Oil Palm Empty Fruit Bunch Fiber Lignin by Chemical and Enzymatic Methods

Directory of Open Access Journals (Sweden)

Pei-Ling Tang

2015-01-01

Full Text Available In this study, oil palm empty fruit bunch (OPEFBF was pretreated with alkali, and lignin was extracted for further degradation into lower molecular weight phenolic compounds using enzymes and chemical means. Efficiency of monomeric aromatic compounds production from OPEFBF lignin via chemical (nitrobenzene versus oxygen and enzymatic [cutinase versus manganese peroxidase (MnP] approaches was investigated. The effects of sodium hydroxide concentration (2, 5, and 10% wt. and reaction time (30, 90, and 180 minutes on the yield of aromatic compounds were studied. The results obtained indicated that nitrobenzene oxidation produced the highest yield (333.17±49.44 ppm hydroxybenzoic acid, 5.67±0.25 ppm p-hydroxybenzaldehyde, 25.57±1.64 ppm vanillic acid, 168.68±23.23 ppm vanillin, 75.44±6.71 ppm syringic acid, 815.26±41.77 ppm syringaldehyde, 15.21±2.19 ppm p-coumaric acid, and 44.75±3.40 ppm ferulic acid, among the tested methods. High sodium hydroxide concentration (10% wt. was needed to promote efficient nitrobenzene oxidation. However, less severe oxidation condition was preferred to preserve the hydroxycinnamic acids (p-coumaric acid and ferulic acid. Cutinase-catalyzed hydrolysis was found to be more efficient than MnP-catalyzed oxidation in the production of aromatic compounds. By hydrolyzed 8% wt. of lignin with 0.625 mL cutinase g−1 lignin at pH 8 and 55°C for 24 hours, about 642.83±14.45 ppm hydroxybenzoic acid, 70.19±3.31 ppm syringaldehyde, 22.80±1.04 ppm vanillin, 27.06±1.20 ppm p-coumaric acid, and 50.19±2.23 ppm ferulic acid were produced.

Effects of the resistivity profile on the formation of a reversed configuration and single helicity states in compressible simulations of the reversed-field pinch

International Nuclear Information System (INIS)

Onofri, M.; Malara, F.

2013-01-01

Compressible magnetohydrodynamics simulations of the reversed-field pinch (RFP) are presented. Previous simulations of the RFP, including density and pressure evolution, showed that a stationary state with a reversed toroidal magnetic field could not be obtained, contrary to the results produced with numerical codes neglecting density and pressure dynamics. The simulations described in the present paper show that including density and pressure evolution, a stationary RFP configuration can be obtained if the resistivity has a radial profile steeply increasing close to the wall. Such resistivity profile is more realistic than a uniform resistivity, since the temperature at the wall is lower than in the plasma core

Studies on mycobacterium tuberculosis sensitivity test by using the method of rapid radiometry with appendixes of clinical results

International Nuclear Information System (INIS)

Yang Yongqing; Jiang Yimin; Lu Wendong; Zhu Rongen

1987-01-01

Three standard strains of mycobacterium tuberculosis (H 37 RV-fully sensitive, SM-R1000 μg/ml, RFP-R 100 μg/ml) were tested with 10 concentration of 5 antitubercular agent, INH, SM, PAS, RFP and EB. 114 isolates of mycobacterium tuberculosis taken from patients were tested with INH, PAS, SM and RFP. They were agreed with the results of standard Lowenstein-Jensen method in 81.7%. 82% of the isolate test were completed within 5 days. The method may be used in routine clinical work. The liquid media prepared by authors do not require human serum albumin and it is less expensive and readily available

The anchorless adhesin Eap (extracellular adherence protein) from Staphylococcus aureus selectively recognizes extracellular matrix aggregates but binds promiscuously to monomeric matrix macromolecules.

Science.gov (United States)

Hansen, Uwe; Hussain, Muzaffar; Villone, Daniela; Herrmann, Mathias; Robenek, Horst; Peters, Georg; Sinha, Bhanu; Bruckner, Peter

2006-05-01

Besides a number of cell wall-anchored adhesins, the majority of Staphylococcus aureus strains produce anchorless, cell wall-associated proteins, such as Eap (extracellular adherence protein). Eap contains four to six tandem repeat (EAP)-domains. Eap mediates diverse biological functions, including adherence and immunomodulation, thus contributing to S. aureus pathogenesis. Eap binding to host macromolecules is unusually promiscuous and includes matrix or matricellular proteins as well as plasma proteins. The structural basis of this promiscuity is poorly understood. Here, we show that in spite of the preferential location of the binding epitopes within triple helical regions in some collagens there is a striking specificity of Eap binding to different collagen types. Collagen I, but not collagen II, is a binding substrate in monomolecular form. However, collagen I is virtually unrecognized by Eap when incorporated into banded fibrils. By contrast, microfibrils containing collagen VI as well as basement membrane-associated networks containing collagen IV, or aggregates containing fibronectin bound Eap as effectively as the monomeric proteins. Therefore, Eap-binding to extracellular matrix ligands is promiscuous at the molecular level but not indiscriminate with respect to supramolecular structures containing the same macromolecules. In addition, Eap bound to banded fibrils after their partial disintegration by matrix-degrading proteinases, including matrix metalloproteinase 1. Therefore, adherence to matrix suprastructures by S. aureus can be supported by inflammatory reactions.

Plasma-column instabilities in a reversed-field pinch without a shell

Energy Technology Data Exchange (ETDEWEB)

Schmid, P.G.

1988-01-01

Plasma column instabilities in a Reversed Field Pinch (RFP) without a shell were investigated in the Colorado Reversatron RFP. The Reversatron RFP (aspect ration R/a = 50 cm/8cm) is a toroidal plasma containment device consisting of a vacuum chamber, a thick conducting shell, modular shells, magnetic field producing coils and diagnostics to characterize the plasma. RFP discharges were set up in the Reversatron in three different experimental configurations: with a thick conducting shell, with a modular shell and with no shell. In two of the configurations, a shell enclosed the plasma column to provide some plasma stability. A vertical magnetic field provided equilibrium in experiments without a shell. Data from discharges without a shell indicated that the plasma duration was greatly reduced and the plasma resistance increased compared to the discharges with a thick shell. Plasma position probes indicated large plasma centriod displacements corresponding to a n = 1 and a n = 3 kink coincident with the peak of the plasma current and the start of a discharge termination phase. The modular shell lengthened the discharge duration and lowered the plasma resistance to values intermediate between the plasma with a thick shell and the plasma with no shell. The modular shell suppressed the large plasma column displacements observed in the RFP plasma without a shell.

IPP partnerships : forging Canada's wind industry

International Nuclear Information System (INIS)

Magee, J.T.

2006-01-01

Following the request for proposal (RFP)-fueled explosion in renewable energy source development over the past 3 years, independent power producer (IPP) financial expertise has become critical to successful project implementation. This presentation reviewed aspects of Canadian wind market related to independent power producer (IPP) investment activity. A map of major Canadian wind farms operational at the end of 2005 was provided, as well as a map of provinces with utility RFP activity. Planned projects were listed by MW. Recent provincial RFP winners were listed. It was suggested that project execution experience and financial security were highly valued in the recent Hydro-Quebec and Ontario RFP solicitations. Developer consolidation and scaling is likely to continue as large wind IPPs are in the best position to maximize efficiency gains in project economics. Financial strength will continue to be key to winning provincial wind RFPs, and Quebec and Ontario RFPs are likely to be dominated by players with balance sheet security. Smaller developers are losing their competitive position in RFP negotiations as more national and international energy giants move into the wind industry. The consolidation trend is ramping up quickly, and may be closing the window in which international investors can find acquisition targets with experience and viable projects. It was concluded that while small developers with robust projects still exist, they are being acquired at a rapid rate. refs., tabs., figs

Radio frequency wave experiments on the MST reversed field pinch

International Nuclear Information System (INIS)

Forest, C.B.; Chattopadhyay, P.K.; Nornberg, M.D.; Prager, S.C.; Thomas, M.A.; Harvey, R.W.; Ram, A.K.

1999-04-01

Experiments, simulations, and theory all indicate that the magnetic fluctuations responsible for the poor confinement in the reversed field pinch (RFP) can be controlled by altering the radial profile of the current density. The magnetic fluctuations in the RFP are due to resistive MHD instabilities caused by current profile peaking; thus confinement in the RFP is ultimately the result of a misalignment between inductively driven current profiles and the stable current profiles characteristic of the Taylor state. If a technique such as rf current drive can be developed to non-inductively sustain a Taylor state (a current profile linearly stable to all tearing modes), the confinement of the RFP and its potential as a reactor concept are likely to increase. Whether there is a self-consistent path from poor confinement to greatly improved confinement through current profile modification is an issue for future experiments to address if and only if near term experiments can demonstrate: (1) coupling to and the propagation of rf waves in RFP plasmas, (2) efficient current drive, and (3) control of the power deposition which will make it possible to control the current profile. In this paper, modeling results and experimental plans are presented for two rf experiments which have the potential of satisfying these three goals: high-n parallel lower hybrid (LH) waves and electron Bernstein waves (EBWs)

Survival, growth performance and immune capacity of the juvenile lined seahorse Hippocampus erectus fed with rifampicin-treated copepods.

Science.gov (United States)

Lin, Tingting; Wang, Changbo; Liu, Xin; Gao, Fen; Xiao, Dongxue; Zhang, Dong; Zhu, Xialian

2017-06-19

Gastrointestinal disease is one of the most serious diseases in cultured seahorse juveniles. Treatment with antimicrobials of live food (i.e. copepods and Artemia) that is used to feed the juveniles may be a promising measure to alleviate the occurrence of gastrointestinal disease. However, relevant investigations are rare. In the present study, we first investigated the antimicrobial efficacies on bacteria within copepods that were treated with 4 antimicrobials, including 3 antibiotics (i.e. enrofloxacin hydrochloride, oxytetracycline and rifampicin [RFP]) that are approved for use in aquaculture and 1 disinfectant (i.e. povidone iodine). We then assessed the effects of copepods treated with the antimicrobial that had the best antimicrobial efficacy on survival, growth performance and immune capacity of juvenile lined seahorses Hippocampus erectus. The results showed that RFP had the best antimicrobial efficacy on both Pseudoalteromonas spp. and Vibrio spp., 2 dominant bacteria with potential pathogenicity within the copepods; the proper concentration of RFP was 6 mg l-1. Moreover, H. erectus juveniles fed with RFP-treated copepods demonstrated an improved survivorship and immune capacity and had a lower abundance of pathogenic bacteria within their gastrointestinal tracts compared to juveniles fed with untreated copepods. These results suggest that treating live food with RFP is a potential measure for reducing the incidence of gastrointestinal disease in seahorse juveniles.

Plasma-column instabilities in a reversed-field pinch without a shell

International Nuclear Information System (INIS)

Schmid, P.G.

1988-01-01

Plasma column instabilities in a Reversed Field Pinch (RFP) without a shell were investigated in the Colorado Reversatron RFP. The Reversatron RFP (aspect ration R/a = 50 cm/8cm) is a toroidal plasma containment device consisting of a vacuum chamber, a thick conducting shell, modular shells, magnetic field producing coils and diagnostics to characterize the plasma. RFP discharges were set up in the Reversatron in three different experimental configurations: with a thick conducting shell, with a modular shell and with no shell. In two of the configurations, a shell enclosed the plasma column to provide some plasma stability. A vertical magnetic field provided equilibrium in experiments without a shell. Data from discharges without a shell indicated that the plasma duration was greatly reduced and the plasma resistance increased compared to the discharges with a thick shell. Plasma position probes indicated large plasma centriod displacements corresponding to a n = 1 and a n = 3 kink coincident with the peak of the plasma current and the start of a discharge termination phase. The modular shell lengthened the discharge duration and lowered the plasma resistance to values intermediate between the plasma with a thick shell and the plasma with no shell. The modular shell suppressed the large plasma column displacements observed in the RFP plasma without a shell

50 CFR 648.21 - Procedures for determining initial annual amounts.

Science.gov (United States)

2010-10-01

... and economic factors. The IOY is composed of a RQ and DAH. RQ will be based on requests for research... equal to ABC. (iii) IOY is composed of RQ, DAH, and TALFF. RQ will be based on requests for research... the RFP. (i) Each panel member will recommend which research proposals should be authorized to utilize...

MANPRINT Handbook for RFP Development

Science.gov (United States)

1990-06-01

a way as to provide ease of movement and transportation (i.e., on a wheeled stand, or with integral casters ). C.4.8 MANPRINT Program. C.4.8.1...constructed. Th6 propos,-, -Huma,. Elgi-ee"rin- "ro( . specify mockups requiring procuring activity approval and modification to -reflect changes. The... modifications to the regulations the Government is evoking. Specifically, the DSL will: (a) Identify by number, revision, title and issuance date all directly

REQUEST FOR PROPOSAL (RFP) Amendment

International Development Research Centre (IDRC) Digital Library (Canada)

Jayne Bassett

2013-08-01

Aug 1, 2013 ... HEAD OFFICE / SIÈGE : 150 Kent Street / 150, rue Kent PO Box / CP 8500 Ottawa ON Canada K1G 3H9 ... Areas of Study. It is understood that the applicant must submit a comprehensive and detailed proposal in one, two, or three areas of study, which include: public health, agriculture and food ...

Optimization of a simple technique for preparation of monodisperse poly(lactide-co-glycolide) nanospheres

Energy Technology Data Exchange (ETDEWEB)

Ito, Fuminori, E-mail: fuminoito@spice.ocn.ne.jp [Tokyo Metropolitan University, Department of Applied Chemistry, Graduate School of Urban Environmental Sciences (Japan)

2016-09-15

In this study, we report the optimization of a solvent evaporation technique for preparing monodisperse poly-(lactide-co-glycolide) (PLGA) nanospheres, from a mixture of solvents composed of ethanol and PVA solution. Various experimental conditions were investigated in order to control the particle size and size distribution of the nanospheres. In addition, nanospheres containing rifampicin (RFP, an antituberculosis drug), were prepared using PLGA of various molecular weights, to study the effects of RFP as a model hydrophobic drug. The results showed that a higher micro-homogenizer stirring rate facilitated the preparation of monodisperse PLGA nanospheres with a low coefficient of variation (~20 %), with sizes below 200 nm. Increasing the PLGA concentration from 0.1 to 0.5 g resulted in an increase in the size of the obtained nanospheres from 130 to 174 nm. The molecular weight of PLGA had little effect on the particle sizes and particle size distributions of the nanospheres. However, the drug loading efficiencies of the obtained RFP/PLGA nanospheres decreased when the molecular weight of PLGA was increased. Based on these experiments, an optimized technique was established for the preparation of monodisperse PLGA nanospheres, using the method developed by the authors.Graphical Abstract.

A one-dimensional plasma and impurity transport model for reversed field pinches

International Nuclear Information System (INIS)

Veerasingam, R.

1991-11-01

In this thesis a one-dimensional (1-D) plasma and impurity transport model is developed to address issues related to impurity behavior in Reversed Field Pinch (RFP) fusion plasmas. A coronal non-equilibrium model is used for impurities. The impurity model is incorporated into an existing one dimensional plasma transport model creating a multi-species plasma transport model which treats the plasma and impurity evolution self-consistently. Neutral deuterium particles are treated using a one-dimensional (slab) model of neutral transport. The resulting mode, RFPBI, is then applied to existing RFP devices such as ZT-40M and MST, and also to examine steady state behavior of ZTH based on the design parameters. A parallel algorithm for the impurity transport equations is implemented and tested to determine speedup and efficiency

The physics of reversed-field pinch profile sustainment

International Nuclear Information System (INIS)

Moses, R.W.

1985-01-01

A description of the Reversed-Field Pinch (RFP) is given. There is experimental evidence that indicates that an RFP dynamo effect sustains field reversal in steady state. Three sustainment mechanisms are reviewed: the MHD model, the tangled discharge model, and the kinetic dynamo model. The relationship of these models to each another is discussed briefly

Start-up assist by magnetized plasma flow injection in TPE-RX reversed-field pinch

Energy Technology Data Exchange (ETDEWEB)

Asai, T. [College of Science and Technology, Nihon University, 1-8 Kanda-Surugadai, Chiyoda-ku, Tokyo 101-8308 (Japan)]. E-mail: asai@phys.cst.nihon-u.ac.jp; Nagata, M. [Graduate School of Engineering, University of Hyogo, Himeji (Japan); Koguchi, H. [National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba (Japan); Hirano, Y. [National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba (Japan); Sakakita, H. [National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba (Japan); Yambe, K. [National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba (Japan); Kiyama, S. [National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba (Japan)

2006-11-15

A reversed-field pinch (RFP) start-up assisted by a magnetized plasma flow injection was demonstrated for the first time on a TPE-RX machine. This sequence of experiments aimed to establish a new method of ionization, gas-fill and helicity injection in the start-up phase of an RFP. In this start-up method, magnetized and well-ionized plasma is formed by a magnetized coaxial plasma gun and injected into the torus chamber as an initial pre-ionized plasma for RFP formation. In the initial experiments, attenuated density pump-out and comparatively slow decay of the toroidal flux and plasma current were observed as evidence of its being an effective start-up method.

The evolution of configuration from q > 1 to q < 1

International Nuclear Information System (INIS)

Zhang Peng

1993-06-01

The evolution of configuration from an initial state of tokamak-like plasma to RFP (reversed field pinch) along the trajectory of minimum energy state is studied. the high plasma current allowed in a RFP is expected to be sufficient to heat the plasma to ignition without the need of auxiliary neutral-beam or radio-frequency heating

A red fluorescent nude mouse model of human endometriosis: advantages of a non-invasive imaging method.

Science.gov (United States)

Wang, Ningning; Hong, Shanshan; Tan, Jinfeng; Ke, Peiqi; Liang, Lili; Fei, Hui; Liu, Bin; Liu, Liqun; Liu, Yongdong; Yu, Bingjun

2014-05-01

To establish red fluorescent human endometriosis lesions in a nude mouse model and dynamically and non-invasively to compare intraperitoneal and subcutaneous injection models. Primary cultures of endometrial stromal cells (ESCs) and epithelial cells (EECs) isolated from 24 patients with a normal uterine cavity were transfected with 2.5×10(8) (Group 1) and 1.25×10(8) (Group 2) plaque-forming units (PFU) of adenovirus encoding red fluorescent protein (Ad-RFP). Transfection efficiencies, fluorescence intensity and apoptosis rate of the two types of cells were compared in vitro. A mixture of 2.5×10(8) PFU Ad-RFP-infected approximately 400 EECs cell mass and 2×10(6) ESCs for 36h was injected individually into 24 female nude mice subcutaneously (Group A) or intraperitoneally (Group B). From Day 5 after injection, an in vivo imaging system (IVIS) was used to non-invasively observe and compare the lesions of the two groups every week until Day 33. Specifically, the fluorescent intensity, positive rates, persistence time and lesion weight in the implanted human endometriosis lesions were compared. A parametric Student's t-test and two-way analysis of variance were used for statistical analysis. Compared with 1.25×10(8) PFU RFP, a titre of 2.5×10(8) PFU RFP ESCs and EECs incubated for 36h exhibited higher transfection efficiencies and higher fluorescence intensities in vitro. In vivo imaging of the fluorescent human endometriosis lesions originating from an RFP titre of 2.5×10(8) PFU showed that the intensity and lesion weight in Group A were significantly higher than in Group B. However, the two groups had the same RFP-positive rates and fluorescence persistence. The structure of each lesion was evaluated by immunohistochemistry to confirm its human endometrial origin. The red fluorescent human endometriosis model established by subcutaneously injecting 2.5×10(8) PFU RFP-transfected stromal cells and epithelial cells into nude mice had a higher fluorescent positive

Novel DDR Processing of Corn Stover Achieves High Monomeric Sugar Concentrations from Enzymatic Hydrolysis (230 g/L) and High Ethanol Concentration (10% v/v) During Fermentation

Energy Technology Data Exchange (ETDEWEB)

Chen, Xiaowen; Jennings, Ed; Shekiro, Joe; Kuhn, Erik M.; O' Brien, Marykate; Wang, Wei; Schell, Daniel J.; Himmel, Mike; Elander, Richard T.; Tucker, Melvin P.

2015-04-03

Distilling and purifying ethanol, butanol, and other products from second and later generation lignocellulosic biorefineries adds significant capital and operating cost for biofuels production. The energy costs associated with distillation affects plant gate and life cycle analysis costs. Lower titers in fermentation due to lower sugar concentrations from pretreatment increase both energy and production costs. In addition, higher titers decrease the volumes required for enzymatic hydrolysis and fermentation vessels. Therefore, increasing biofuels titers has been a research focus in renewable biofuels production for several decades. In this work, we achieved over 200 g/L of monomeric sugars after high solids enzymatic hydrolysis using the novel deacetylation and disc refining (DDR) process on corn stover. The high sugar concentrations and low chemical inhibitor concentrations from the DDR process allowed ethanol titers as high as 82 g/L in 22 hours, which translates into approximately 10 vol% ethanol. To our knowledge, this is the first time that 10 vol% ethanol in fermentation derived from corn stover without any sugar concentration or purification steps has been reported. Techno-economic analysis shows the higher titer ethanol achieved from the DDR process could significantly reduce the minimum ethanol selling price from cellulosic biomass.

State of insulin self-association does not affect its absorption from the pulmonary route.

Science.gov (United States)

Hussain, Alamdar; Ahsan, Fakhrul

2005-06-01

This study is designed to compare and contrast the pulmonary absorption profiles of monomeric and hexameric insulin in the presence or absence of ethylene diamine tetraacetic acid (EDTA) or n-tetradecyl-beta-d-maltoside (TDM). The pulmonary absorption of two forms of insulin was studied by monitoring the changes in plasma insulin and glucose levels after intratracheal administration of monomeric or hexameric insulin into anesthetized rodents. EDTA or TDM was added to the formulation in order to evaluate if either of these agents has effects on the rate and extent of pulmonary absorption of monomeric and hexameric insulin. The biochemical changes that may occur after acute administration of TDM-based formulation have also been investigated by estimating lung injury markers in bronchoalveolar lavage fluid. A dose-dependent increase in the plasma insulin and decrease in plasma glucose levels was observed when increasing concentrations of hexameric or monomeric insulin were administered via the pulmonary route. Pulmonary administration of monomeric and hexameric insulin produced comparable absorption profiles in the presence or absence of EDTA or TDM. The bronchoalveolar lavage fluid analysis did not show differences in the levels of injury markers produced in TDM-treated rats and that produced in saline-treated rats, indicating no evidence for adverse effects of TDM in these short-term studies. Overall, in terms of rapidity of action and efficacy to reduce blood sugar, monomeric insulin did not provide advantages over hexameric insulin when administered via the pulmonary route.

The TITAN Reversed-Field Pinch fusion reactor study

International Nuclear Information System (INIS)

1988-03-01

The TITAN Reversed-Field Pinch (RFP) fusion reactor study is a multi-institutional research effort to determine the technical feasibility and key developmental issues of an RFP fusion reactor, especially at high power density, and to determine the potential economics, operations, safety, and environmental features of high-mass-power-density fusion systems. The TITAN conceptual designs are DT burning, 1000 MWe power reactors based on the RFP confinement concept. The designs are compact, have a high neutron wall loading of 18 MW/m 2 and a mass power density of 700 kWe/tonne. The inherent characteristics of the RFP confinement concept make fusion reactors with such a high mass power density possible. Two different detailed designs have emerged: the TITAN-I lithium-vanadium design, incorporating the integrated-blanket-coil concept; and the TITAN-II aqueous loop-in-pool design with ferritic steel structure. This report contains a collection of 16 papers on the results of the TITAN study which were presented at the International Symposium on Fusion Nuclear Technology. This collection describes the TITAN research effort, and specifically the TITAN-I and TITAN-II designs, summarizing the major results, the key technical issues, and the central conclusions and recommendations. Overall, the basic conclusions are that high-mass power-density fusion reactors appear to be technically feasible even with neutron wall loadings up to 20 MW/m 2 ; that single-piece maintenance of the FPC is possible and advantageous; that the economics of the reactor is enhanced by its compactness; and the safety and environmental features need not to be sacrificed in high-power-density designs. The fact that two design approaches have emerged, and others may also be possible, in some sense indicates the robustness of the general findings

Robust, synergistic regulation of human gene expression using TALE activators.

Science.gov (United States)

Maeder, Morgan L; Linder, Samantha J; Reyon, Deepak; Angstman, James F; Fu, Yanfang; Sander, Jeffry D; Joung, J Keith

2013-03-01

Artificial activators designed using transcription activator-like effector (TALE) technology have broad utility, but previous studies suggest that these monomeric proteins often exhibit low activities. Here we demonstrate that TALE activators can robustly function individually or in synergistic combinations to increase expression of endogenous human genes over wide dynamic ranges. These findings will encourage applications of TALE activators for research and therapy, and guide design of monomeric TALE-based fusion proteins.

Magnetic islands at the field reversal surface in reversed field pinches

International Nuclear Information System (INIS)

Pinsker, R.I.; Reiman, A.H.

1985-09-01

In the reversed field pinch (RFP), magnetic field perturbations having zero poloidal mode number and any toroidal mode number are resonant at the field reversal surface. Such perturbations are a particular threat to the RFP because of their weak radial dependence at low toroidal mode number, and because the toroidal field ripple is essentially of this type. The widths of the resulting islands are calculated in this paper. The self-consistent plasma response is included through the assumption that the plasma relaxes to a Taylor force-free state. The connection with linear tearing mode theory is established for those limits where arbitrarily large islands result from infinitesimal perturbations. Toroidal effects are considered, and application of the theory to RFP experiments is discussed

Recent studies of Reversed-Field Pinch reactors

International Nuclear Information System (INIS)

Hagenson, R.L.; Krakowski, R.A.

1981-01-01

The reactor prognoses of a class of confinement scheme that relies primarily on self-fields induced by axial currents flowing within a plasma column are presented. The primary focus has been placed on the toroidal Reversed-Field Pinch (RFP). At the limit of very large current densities is the gas-embedded Dense Z-Pinch (DZP), a small-radius, linear device. Past conventional RFP reactor designs are reviewed. The extention of these conventional RFP reactors to DD advanced-fuel operation is described. The implications are summarized of operating higher-density, compact RFPs as reactors, wherein the current density rather than physical dimensions are scaled. Lastly, the application of very high current densities supported in a sub-millimeter linear current channel, as embodied in the DZP reactor, is reviewed

FluBreaks: early epidemic detection from Google flu trends.

Science.gov (United States)

Pervaiz, Fahad; Pervaiz, Mansoor; Abdur Rehman, Nabeel; Saif, Umar

2012-10-04

The Google Flu Trends service was launched in 2008 to track changes in the volume of online search queries related to flu-like symptoms. Over the last few years, the trend data produced by this service has shown a consistent relationship with the actual number of flu reports collected by the US Centers for Disease Control and Prevention (CDC), often identifying increases in flu cases weeks in advance of CDC records. However, contrary to popular belief, Google Flu Trends is not an early epidemic detection system. Instead, it is designed as a baseline indicator of the trend, or changes, in the number of disease cases. To evaluate whether these trends can be used as a basis for an early warning system for epidemics. We present the first detailed algorithmic analysis of how Google Flu Trends can be used as a basis for building a fully automated system for early warning of epidemics in advance of methods used by the CDC. Based on our work, we present a novel early epidemic detection system, called FluBreaks (dritte.org/flubreaks), based on Google Flu Trends data. We compared the accuracy and practicality of three types of algorithms: normal distribution algorithms, Poisson distribution algorithms, and negative binomial distribution algorithms. We explored the relative merits of these methods, and related our findings to changes in Internet penetration and population size for the regions in Google Flu Trends providing data. Across our performance metrics of percentage true-positives (RTP), percentage false-positives (RFP), percentage overlap (OT), and percentage early alarms (EA), Poisson- and negative binomial-based algorithms performed better in all except RFP. Poisson-based algorithms had average values of 99%, 28%, 71%, and 76% for RTP, RFP, OT, and EA, respectively, whereas negative binomial-based algorithms had average values of 97.8%, 17.8%, 60%, and 55% for RTP, RFP, OT, and EA, respectively. Moreover, the EA was also affected by the region's population size

Influence of the environment and phototoxicity of the live cell imaging system at IMP microbeam facility

Science.gov (United States)

Liu, Wenjing; Du, Guanghua; Guo, Jinlong; Wu, Ruqun; Wei, Junzhe; Chen, Hao; Li, Yaning; Zhao, Jing; Li, Xiaoyue

2017-08-01

To investigate the spatiotemporal dynamics of DNA damage and repair after the ion irradiation, an online live cell imaging system has been established based on the microbeam facility at Institute of Modern Physics (IMP). The system could provide a sterile and physiological environment by making use of heating plate and live cell imaging solution. The phototoxicity was investigated through the evaluation of DNA repair protein XRCC1 foci formed in HT1080-RFP cells during the imaging exposure. The intensity of the foci induced by phototoxicity was much lower compared with that of the foci induced by heavy ion hits. The results showed that although spontaneous foci were formed due to RFP exposure during live cell imaging, they had little impact on the analysis of the recruitment kinetics of XRCC1 in the foci induced by the ion irradiation.

Technical Safety Appraisal of the Rocky Flats Plant

Energy Technology Data Exchange (ETDEWEB)

Brown, Blake P.

1989-01-01

This report provides the results of a Technical Safety Appraisal (TSA) of the Rocky Flats Plant (RFP) conducted November 14 to 18 and November 28 to December 9, 1988. This appraisal covered the effectiveness and improvements in the RFP safety program across the site, evaluating progress to date against standards of accepted practice. The appraisal included coverage of the timeliness and effectiveness of actions taken in response to the recommendations/concerns in three previous Technical Safety Appraisals (TSAs) of RFP Bldg. 707 conducted in July 1986, Bldgs. 771/774 conducted in October/November 1986, and Bldgs. 776/777 conducted in January/February 1988. Results of this appraisal are given in Section IV for each of 14 technical safety areas at RFP. These results include a discussion, conclusions and any new safety concerns for each technical safety area. Appendix A contains a description of the system for categorizing concerns, and the concerns are tabulated in Appendix B. Appendix C reports on the evaluation of the contractor's actions and the current status of each of the 230 recommendations and concerns contained in the three previous TSA reports.

Identification of active Plasmodium falciparum calpain to establish screening system for Pf-calpain-based drug development

Directory of Open Access Journals (Sweden)

Soh Byoung

2013-02-01

Full Text Available Abstract Background With the increasing resistance of malaria parasites to available drugs, there is an urgent demand to develop new anti-malarial drugs. Calpain inhibitor, ALLN, is proposed to inhibit parasite proliferation by suppressing haemoglobin degradation. This provides Plasmodium calpain as a potential target for drug development. Pf-calpain, a cysteine protease of Plasmodium falciparum, belongs to calpain-7 family, which is an atypical calpain not harboring Ca2+-binding regulatory motifs. In this present study, in order to establish the screening system for Pf-calpain specific inhibitors, the active form of Pf-calpain was first identified. Methods Recombinant Pf-calpain including catalytic subdomain IIa (rPfcal-IIa was heterologously expressed and purified. Enzymatic activity was determined by both fluorogenic substrate assay and gelatin zymography. Molecular homology modeling was carried out to address the activation mode of Pf-calpain in the aspect of structural moiety. Results Based on the measurement of enzymatic activity and protease inhibitor assay, it was found that the active form of Pf-calpain only contains the catalytic subdomain IIa, suggesting that Pf-calpain may function as a monomeric form. The sequence prediction indicates that the catalytic subdomain IIa contains all amino acid residues necessary for catalytic triad (Cys-His-Asn formation. Molecular modeling suggests that the Pf-calpain subdomain IIa makes an active site, holding the catalytic triad residues in their appropriate orientation for catalysis. The mutation analysis further supports that those amino acid residues are functional and have enzymatic activity. Conclusion The identified active form of Pf-calpain could be utilized to establish high-throughput screening system for Pf-calpain inhibitors. Due to its unique monomeric structural property, Pf-calpain could be served as a novel anti-malarial drug target, which has a high specificity for malaria parasite

Time-resolved observation of discrete and continuous MHD dynamo in the reversed-field pinch edge

International Nuclear Information System (INIS)

Ji, H.; Almagri, A.F.; Prager, S.C.; Sarff, J.S.

1994-01-01

We report the first experimental verification of the MHD dynamo in the RFP. A burst of magnetohydrodynamic (MHD) dynamo electric field is observed during the sawtooth crash, followed by an increase in the local parallel current in the MST RFP edge. By measuring each term, the parallel MHD mean-field Ohm's law is observed to hold within experimental error bars both between and during sawtooth crashes

Autonomous Star Tracker Algorithms

DEFF Research Database (Denmark)

Betto, Maurizio; Jørgensen, John Leif; Kilsgaard, Søren

1998-01-01

Proposal, in response to an ESA R.f.P., to design algorithms for autonomous star tracker operations.The proposal also included the development of a star tracker breadboard to test the algorithms performances.......Proposal, in response to an ESA R.f.P., to design algorithms for autonomous star tracker operations.The proposal also included the development of a star tracker breadboard to test the algorithms performances....

The monomeric GTPase RabA2 is required for progression and maintenance of membrane integrity of infection threads during root nodule symbiosis.

Science.gov (United States)

Dalla Via, Virginia; Traubenik, Soledad; Rivero, Claudio; Aguilar, O Mario; Zanetti, María Eugenia; Blanco, Flavio Antonio

2017-04-01

Progression of the infection canal that conducts rhizobia to the nodule primordium requires a functional Rab GTPase located in Golgi/trans-Golgi that also participate in root hair polar growth. Common bean (Phaseolus vulgaris) symbiotically associates with its partner Rhizobium etli, resulting in the formation of root nitrogen-fixing nodules. Compatible bacteria can reach cortical cells in a tightly regulated infection process, in which the specific recognition of signal molecules is a key step to select the symbiotic partner. In this work, we show that RabA2, a monomeric GTPase from common bean, is required for the progression of the infection canal, referred to as the infection thread (IT), toward the cortical cells. Expression of miss-regulated mutant variants of RabA2 resulted in an increased number of abortive infection events, including bursting of ITs and a reduction in the number of nodules. Nodules formed in these plants were small and contained infected cells with disrupted symbiosome membranes, indicating either early senescence of these cells or defects in the formation of the symbiosome membrane during bacterial release. RabA2 localized to mobile vesicles around the IT, but mutations that affect GTP hydrolysis or GTP/GDP exchange modified this localization. Colocalization of RabA2 with ArfA1 and a Golgi marker indicates that RabA2 localizes in Golgi stacks and the trans-Golgi network. Our results suggest that RabA2 is part of the vesicle transport events required to maintain the integrity of the membrane during IT progression.

Controlled catalytic and thermal sequential pyrolysis and hydrolysis of phenolic resin containing waste streams to sequentially recover monomers and chemicals

Science.gov (United States)

Chum, H.L.; Evans, R.J.

1992-08-04

A process is described for using fast pyrolysis in a carrier gas to convert a waste phenolic resin containing feedstreams in a manner such that pyrolysis of said resins and a given high value monomeric constituent occurs prior to pyrolyses of the resins in other monomeric components therein comprising: selecting a first temperature program range to cause pyrolysis of said resin and a given high value monomeric constituent prior to a temperature range that causes pyrolysis of other monomeric components; selecting, if desired, a catalyst and a support and treating said feedstreams with said catalyst to effect acid or basic catalyzed reaction pathways to maximize yield or enhance separation of said high value monomeric constituent in said first temperature program range to utilize reactive gases such as oxygen and steam in the pyrolysis process to drive the production of specific products; differentially heating said feedstreams at a heat rate within the first temperature program range to provide differential pyrolysis for selective recovery of optimum quantity of said high value monomeric constituent prior to pyrolysis of other monomeric components therein; separating said high value monomeric constituent; selecting a second higher temperature program range to cause pyrolysis of a different high value monomeric constituent of said phenolic resins waste and differentially heating said feedstreams at said higher temperature program range to cause pyrolysis of said different high value monomeric constituent; and separating said different high value monomeric constituent. 11 figs.

Plasma performance and scaling laws in the RFX-mod reversed-field pinch experiment

International Nuclear Information System (INIS)

Innocente, P.; Alfier, A.; Canton, A.; Pasqualotto, R.

2009-01-01

The large range of plasma currents (I p = 0.2-1.6 MA) and feedback-controlled magnetic boundary conditions of the RFX-mod experiment make it well suited to performing scaling studies. The assessment of such scaling, in particular those on temperature and energy confinement, is crucial both for improving the operating reversed-field pinch (RFP) devices and for validating the RFP configuration as a candidate for the future fusion reactors. For such a purpose scaling laws for magnetic fluctuations, temperature and energy confinement have been evaluated in stationary operation. RFX-mod scaling laws have been compared with those obtained from other RFP devices and numerical simulations. The role of the magnetic boundary has been analysed, comparing discharges performed with different active control schemes of the edge radial magnetic field.

Palm oil based polymer materials obtained by ROMP: study by low field NMR

International Nuclear Information System (INIS)

Fernandes, Henrique; Azevedo, Eduardo R. de; Lima-Neto, Benedito S.

2015-01-01

Aiming to study and develop new materials synthesized from sustainable sources, several polymers were prepared using in its monomeric composition, different amounts of NPO (Norbornenyl Palm Oil) monomer. This monomer was developed based on a vegetable oil rather produced in northern Brazil, the Palm Oil. Since this oil have a low content of unsaturation, its use in developing new monomer for ROMP (Ring-Opening Metathesis Polymerization) is not exploited. In this regard, polymeric materials were obtained using the NOP and both the reaction process and the resulting products were analyzed by Nuclear Magnetic Resonance in the time domain (TD-NMR) at low magnetic field. (author)

50 CFR 648.207 - Herring Research Set-Aside (RSA).

Science.gov (United States)

2010-10-01

... 50 Wildlife and Fisheries 8 2010-10-01 2010-10-01 false Herring Research Set-Aside (RSA). 648.207... Measures for the Atlantic Herring Fishery § 648.207 Herring Research Set-Aside (RSA). (a) NMFS shall... authorized to utilize RSA, based on the selection criteria described in the RFP. (d) NMFS shall consider each...

Solid structures of the stepwise self-assembled copillar[5]arene-based supramolecular polymers

International Nuclear Information System (INIS)

Park, Yeon Sil; Hwang, Seong Min; Shin, Jae Yeon; Paek, Kyung Soo

2016-01-01

Development of supramolecular polymer has attracted much interest because of their interesting properties such as stimuli-responsiveness, recycling, self-healing and degradability, and their consequential applications. The essential feature of this class of polymers is the self-assembly of discrete monomeric subunits via non-covalent interactions or dynamic covalent bonds. Among the many monomeric subunits, pillar[n]arenes have been ideal building blocks for the fabrication of polymeric supramolecules because of their intrinsic characteristics. The ring-shaped morphologies in supramolecular polymer P are probably due to the tendency of the end-to-end connection in the solid state of long flexible supramolecular chains. The size increase of nano-rings as the stepwise addition increases might be due to the fact that the linear supramolecular polymer P in solution seems to be maintained until the nano-ring formation by solidification

Solid structures of the stepwise self-assembled copillar[5]arene-based supramolecular polymers

Energy Technology Data Exchange (ETDEWEB)

Park, Yeon Sil; Hwang, Seong Min; Shin, Jae Yeon; Paek, Kyung Soo [Dept. of Chemistry, Soongsil University, Seoul (Korea, Republic of)

2016-10-15

Development of supramolecular polymer has attracted much interest because of their interesting properties such as stimuli-responsiveness, recycling, self-healing and degradability, and their consequential applications. The essential feature of this class of polymers is the self-assembly of discrete monomeric subunits via non-covalent interactions or dynamic covalent bonds. Among the many monomeric subunits, pillar[n]arenes have been ideal building blocks for the fabrication of polymeric supramolecules because of their intrinsic characteristics. The ring-shaped morphologies in supramolecular polymer P are probably due to the tendency of the end-to-end connection in the solid state of long flexible supramolecular chains. The size increase of nano-rings as the stepwise addition increases might be due to the fact that the linear supramolecular polymer P in solution seems to be maintained until the nano-ring formation by solidification.

Heat transport modelling in EXTRAP T2R

Science.gov (United States)

Frassinetti, L.; Brunsell, P. R.; Cecconello, M.; Drake, J. R.

2009-02-01

A model to estimate the heat transport in the EXTRAP T2R reversed field pinch (RFP) is described. The model, based on experimental and theoretical results, divides the RFP electron heat diffusivity χe into three regions, one in the plasma core, where χe is assumed to be determined by the tearing modes, one located around the reversal radius, where χe is assumed not dependent on the magnetic fluctuations and one in the extreme edge, where high χe is assumed. The absolute values of the core and of the reversal χe are determined by simulating the electron temperature and the soft x-ray and by comparing the simulated signals with the experimental ones. The model is used to estimate the heat diffusivity and the energy confinement time during the flat top of standard plasmas, of deep F plasmas and of plasmas obtained with the intelligent shell.

Advanced-fuel reversed-field pinch reactor (RFPR)

International Nuclear Information System (INIS)

Hagenson, R.L.; Krakowski, R.A.

1981-10-01

The utilization of deuterium-based fuels offers the potential advantages of greater flexibility in blanket design, significantly reduced tritium inventory, potential reduction in radioactivity level, and utilization of an inexhaustible fuel supply. The conventional DT-fueled Reversed-Field Pinch Reactor (RFPR) designs are reviewed, and the recent extension of these devices to advanced-fuel (catalyzed-DD) operation is presented. Attractive and economically competitive DD/RFPR systems are identified having power densities and plasma parameters comparable to the DT systems. Converting an RFP reactor from DT to DD primarily requires increasing the magnetic field levels a factor of two, still requiring only modest magnet coil fields (less than or equal to 4 T). When compared to the mainline tokamak, the unique advantages of the RFP (e.g., high beta, low fields at the coils, high ohmic-heating power densities, unrestricted aspect ratio) are particularly apparent for the utilization of advanced fuels

Self-consistent equilibrium in a cylindrical, dissipative reverse field pinch

International Nuclear Information System (INIS)

Guo, S.C.; Paccagnella, R.

1994-01-01

One of the authors (C.L.S.) recently proposed a dissipative model to self-consistently solve the equilibrium problem in a free-boundary plasma column under cylindrical symmetry. In the present paper, on one hand the problem is strongly specialized to circular symmetry and to Ohm's and Fourier's laws without off-diagonal contributions; on the other hand, it is generalized by adding a dynamo effective electric field E d in Ohm's law, based on the standard turbulent model. This seems appropriate enough to study RFP equilibria, since it is well known that a stationary and cylindrically symmetric RFP is incompatible with a classical Ohm's law. Reasonably, only numerical solutions are expected to be accessible in general; but the further simplified problem with scalar and constant electric resistivity and constant dynamo coefficient α (E d =αB) can be solved analytically by elementary means. (author) 4 refs., 2 figs

A collimated neutron detector for RFP plasmas in MST

Energy Technology Data Exchange (ETDEWEB)

Capecchi, W. J., E-mail: capecchi@wisc.edu; Anderson, J. K.; Bonofiglo, P. J.; Kim, J.; Sears, S. [University of Wisconsin- Madison, Madison, Wisconsin 53706 (United States)

2016-11-15

The neutron emissivity profile in the Madison Symmetric Torus is being reconstructed through the use of a collimated neutron detector. A scintillator-photomultiplier tube (PMT) system is employed to detect the fusion neutrons with the plasma viewing volume defined by a 55 cm deep, 5 cm diameter aperture. Effective detection of neutrons from the viewing volume is achieved through neutron moderation using 1300 lbs of high density polyethylene shielding, which modeling predicts attenuates the penetrating flux by a factor of 10{sup 4} or more. A broad spectrum of gamma radiation is also present due to the unconfined fusion proton bombardment of the thick aluminum vacuum vessel. A 15 cm cylindrical liquid scintillator of 3.8 cm diameter is used to further increase directional sensitivity. A fast (5 ns rise time) preamplifier and digitization at 500 MHz prevent pulse pile-up even at high count rates (∼10{sup 4}/s). The entire neutron camera system is situated on an adjustable inclining base which provides the differing plasma viewing volumes necessary for reconstruction of the neutron emissivity profile. This profile, directly related to the fast-ion population, allows for an investigation of the critical fast-ion pressure gradient required to destabilize a neutral beam driven Alfvénic mode which has been shown to transport fast ions.

Viscosity of iodinated contrast agents during renal excretion

International Nuclear Information System (INIS)

Jost, Gregor; Lengsfeld, Philipp; Lenhard, Diana C.; Pietsch, Hubertus; Huetter, Joachim; Sieber, Martin A.

2011-01-01

Objective: Modern iodinated non-ionic contrast agents (CAs) can be classified based on their molecular structure into monomeric and dimeric CAs and have at comparable iodine concentrations a different viscosity and osmolality. During their renal excretion, CAs are concentrated in the renal tubuli which might enhance the viscosity difference between monomeric and dimeric CAs. The viscosity of a CA might have an underestimated importance for renal safety, as suggested by recent publications. In this study, we investigated the viscosities of CAs at the concentrations expected to be present in renal tubules. This concentration process was simulated in vitro using dialysis. Furthermore, we investigated urine viscosity and urine flow in rodents after administration of several non-ionic monomeric and dimeric CAs. Materials and methods: To estimate the viscosity of the CAs in vivo, we performed an in vitro dialysis of monomeric and dimeric CAs at various physiological osmolalities of the renal tubulus (290, 400, 500, 700 and 1000 mOsm/kg H 2 O). Following the dialysis, the iodine concentrations and the viscosities of the CAs were determined. Furthermore, to investigate the concentration process in vivo, we measured the urine viscosity and the urine flow in Han Wister rats after the administration of Iopromide, Iohexol, Ioversol, Iomeprol, Iodixanol, and Iosimenol at comparable iodine concentrations. As a control, saline was injected at the same volume. Results: In vitro dialysis of the dimeric CA increased the iodine concentration and strongly increased the viscosity at all tested osmolalities. In contrast, for the monomeric agents an increase in concentration and viscosity was observed only at 700 as well 1000 mOsm/kg H 2 O but to a lesser extent. In summary, dialysis strongly enhanced the viscosity differences between the non-ionic monomeric and dimeric CAs. The administration of dimeric CAs leads to a strong increase in urine viscosity; this was not observed for the

Viscosity of iodinated contrast agents during renal excretion

Energy Technology Data Exchange (ETDEWEB)

Jost, Gregor, E-mail: Gregor.Jost@bayer.com [TRG Diagnostic Imaging, Bayer Schering Pharma AG, Berlin (Germany); Lengsfeld, Philipp, E-mail: Philipp.Lengsfeld@bayer.com [Global Medical Affairs Diagnostic Imaging, Bayer Schering Pharma AG, Berlin (Germany); Lenhard, Diana C., E-mail: Diana.Lenhard@bayer.com [TRG Diagnostic Imaging, Bayer Schering Pharma AG, Berlin (Germany); Pietsch, Hubertus, E-mail: Hubertus.Pietsch@bayer.com [TRG Diagnostic Imaging, Bayer Schering Pharma AG, Berlin (Germany); Huetter, Joachim, E-mail: Joachim.Huetter@bayer.com [TRG Diagnostic Imaging, Bayer Schering Pharma AG, Berlin (Germany); Sieber, Martin A., E-mail: Martin.Sieber@bayer.com [TRG Diagnostic Imaging, Bayer Schering Pharma AG, Berlin (Germany)

2011-11-15

Objective: Modern iodinated non-ionic contrast agents (CAs) can be classified based on their molecular structure into monomeric and dimeric CAs and have at comparable iodine concentrations a different viscosity and osmolality. During their renal excretion, CAs are concentrated in the renal tubuli which might enhance the viscosity difference between monomeric and dimeric CAs. The viscosity of a CA might have an underestimated importance for renal safety, as suggested by recent publications. In this study, we investigated the viscosities of CAs at the concentrations expected to be present in renal tubules. This concentration process was simulated in vitro using dialysis. Furthermore, we investigated urine viscosity and urine flow in rodents after administration of several non-ionic monomeric and dimeric CAs. Materials and methods: To estimate the viscosity of the CAs in vivo, we performed an in vitro dialysis of monomeric and dimeric CAs at various physiological osmolalities of the renal tubulus (290, 400, 500, 700 and 1000 mOsm/kg H{sub 2}O). Following the dialysis, the iodine concentrations and the viscosities of the CAs were determined. Furthermore, to investigate the concentration process in vivo, we measured the urine viscosity and the urine flow in Han Wister rats after the administration of Iopromide, Iohexol, Ioversol, Iomeprol, Iodixanol, and Iosimenol at comparable iodine concentrations. As a control, saline was injected at the same volume. Results: In vitro dialysis of the dimeric CA increased the iodine concentration and strongly increased the viscosity at all tested osmolalities. In contrast, for the monomeric agents an increase in concentration and viscosity was observed only at 700 as well 1000 mOsm/kg H{sub 2}O but to a lesser extent. In summary, dialysis strongly enhanced the viscosity differences between the non-ionic monomeric and dimeric CAs. The administration of dimeric CAs leads to a strong increase in urine viscosity; this was not observed for

Skill Transfer and Virtual Training for IND Response Decision-Making: Models for Government-Industry Collaboration for the Development of Game-Based Training Tools

Science.gov (United States)

2016-05-05

for the player or student. For example, students at the Berkeley School of Music are world-class musicians who are often available to help with game...or smaller gatherings such as the annual Boston Festival of Independent Games (BFIG), are inexpensive, effective means of getting the word out...NUSTL RFP RTS SME artificial intelligence broad agency announcement Boston Festival of Indie Games Chemical Biological Radiological Nuclear

The SIMPSONS project: An integrated Mars transportation system

Science.gov (United States)

Kaplan, Matthew; Carlson, Eric; Bradfute, Sherie; Allen, Kent; Duvergne, Francois; Hernandez, Bert; Le, David; Nguyen, Quan; Thornhill, Brett

In response to the Request for Proposal (RFP) for an integrated transportation system network for an advanced Martian base, Frontier Transportation Systems (FTS) presents the results of the SIMPSONS project (Systems Integration for Mars Planetary Surface Operations Networks). The following topics are included: the project background, vehicle design, future work, conclusions, management status, and cost breakdown. The project focuses solely on the surface-to-surface transportation at an advanced Martian base.

Imaging Nuclear-Cytoplasmic Dynamics in Primary and Metastatic Colon Cancer in Nude Mice.

Science.gov (United States)

Hasegawa, Kosuke; Suetsugu, Atsushi; Nakamura, Miki; Matsumoto, Takuro; Aoki, Hitomi; Kunisada, Takahiro; Bouvet, Michael; Shimizu, Masahito; Hoffman, Robert M

2016-05-01

Colon cancer frequently results in metastasis to the liver, where it becomes the main cause of death. However, the cell cycle in primary tumors and metastases is poorly understood. We developed a mouse model of liver metastasis using the human colon cancer cell line HCT-116, which expresses green fluorescent protein (GFP) in the nucleus and red fluorescent protein (RFP) in the cytoplasm (HCT-116-GFP-RFP). HCT-116 GFP-RFP cells were injected into the spleen of nu/nu nude mice. HCT-116-GFP-RFP cells subsequently formed primary tumors in the spleen, as well as metastatic colonies in the liver and retroperitoneum by 28 days after cell transplantation. Using an Olympus FV1000 confocal microscope, it was possible to clearly image mitosis of the dual-colored colon cancer cells in the primary tumor as well as liver and other metastases. Multi-nucleate cancer cells, in addition to mono-nucleate cancer cells and their mitosis, were observed in the primary tumor and metastasis. Multi-nucleate HCT-116-GFP-RFP cells were also observed after culture of the primary and metastatic tumors. A similar ratio of mono-nucleate, multi-nucleate, and mitotic cells grew from the primary and metastatic tumors in culture, suggesting similarity of the nuclear-cytoplasmic dynamics of primary and metastatic cancer cells, further emphasizing the stochastic nature of metastasis. Our results demonstrate a similar heterogeneity of nuclear-cytoplasmic dynamics within primary tumors and metastases, which may be an important factor in the stochastic nature of metastasis. Copyright© 2016 International Institute of Anticancer Research (Dr. John G. Delinassios), All rights reserved.

Recognized focused practice: Does sub-specialty designation offer value to the neurosurgeon?

Directory of Open Access Journals (Sweden)

Maya A Babu

Full Text Available Vehicles for life-long assessment such as Maintenance of Certification tend to focus on generalist neurosurgical knowledge. However, as neurosurgeons advance in their careers, they tend to narrow their practice and increase volumes in certain specific types of operations. Failing to test the type of procedures most relevant to the practitioner is a lost opportunity to improve the knowledge and practice of the individual neurosurgeon. In this study, we assess the neurosurgical community's appetite for designations of board-recognized Recognized Focused Practice (RFP. We administered a validated, online, confidential survey to 4,899 neurosurgeons (2,435 American Board of Neurological Surgery (ABNS Diplomates participating in MOC, 1,440 Diplomates certified prior to 1999 (grandfathered, and 1,024 retired Diplomates. We received 1,449 responses overall (30% response rate. A plurality of respondents were in practice 11-15 years (18.5%, in private practice (40% and participate in MOC (61%. 49% of respondents felt that a RFP designation would not be helpful. For the 30% who felt that RFP would be helpful, 61.3% felt that it would support recognition by their hospital or practice, it would motivate them to stay current on medical knowledge (53.4%, or it would help attract patients (46.4%;. The most popular suggestions for RFP were Spine (56.2%, Cerebrovascular (62.9%, Pediatrics (64.1%, and Functional/Stereotactic (52%. A plurality of neurosurgeons (35.7% felt that RFP should recognize neurosurgeons with accredited and non-accredited fellowship experience and sub-specialty experience. Ultimately, Recognized Focused Practice may provide value to individual neurosurgeons, but the neurosurgical community shows tepid interest for pursuing this designation.

Recognized focused practice: Does sub-specialty designation offer value to the neurosurgeon?

Science.gov (United States)

Babu, Maya A; Liau, Linda M; Meyer, Fredric B

2017-01-01

Vehicles for life-long assessment such as Maintenance of Certification tend to focus on generalist neurosurgical knowledge. However, as neurosurgeons advance in their careers, they tend to narrow their practice and increase volumes in certain specific types of operations. Failing to test the type of procedures most relevant to the practitioner is a lost opportunity to improve the knowledge and practice of the individual neurosurgeon. In this study, we assess the neurosurgical community's appetite for designations of board-recognized Recognized Focused Practice (RFP). We administered a validated, online, confidential survey to 4,899 neurosurgeons (2,435 American Board of Neurological Surgery (ABNS) Diplomates participating in MOC, 1,440 Diplomates certified prior to 1999 (grandfathered), and 1,024 retired Diplomates). We received 1,449 responses overall (30% response rate). A plurality of respondents were in practice 11-15 years (18.5%), in private practice (40%) and participate in MOC (61%). 49% of respondents felt that a RFP designation would not be helpful. For the 30% who felt that RFP would be helpful, 61.3% felt that it would support recognition by their hospital or practice, it would motivate them to stay current on medical knowledge (53.4%), or it would help attract patients (46.4%;). The most popular suggestions for RFP were Spine (56.2%), Cerebrovascular (62.9%), Pediatrics (64.1%), and Functional/Stereotactic (52%). A plurality of neurosurgeons (35.7%) felt that RFP should recognize neurosurgeons with accredited and non-accredited fellowship experience and sub-specialty experience. Ultimately, Recognized Focused Practice may provide value to individual neurosurgeons, but the neurosurgical community shows tepid interest for pursuing this designation.

Kinematics of Nonlinearly Interacting MHD Instabilities in a Plasma

International Nuclear Information System (INIS)

Hansen, Alexander K.

2000-01-01

Plasmas play host to a wide variety of instabilities. For example, tearing instabilities use finite plasma resistivity to exploit the free energy provided by plasma currents parallel to the magnetic field to alter the magnetic topology of the plasma through a process known as reconnection. These instabilities frequently make themselves known in magnetic confinement experiments such as tokamaks and reversed field pinches (RFPs). In RFP plasmas, in fact, several tearing instabilities (modes) are simultaneously active, and are of large amplitude. Theory predicts that in addition to interacting linearly with magnetic perturbations from outside the plasma, such as field errors or as resistive wall, the modes in the RFP can interact nonlinearly with each other through a three-wave interaction. In the current work investigations of both the linear (external) and nonlinear contributions to the kinematics of the tearing modes in the Madison Symmetric Torus (MST) RFP are reported Theory predicts that tearing modes will respond only to magnetic perturbations that are spatially resonant with them, and was supported by experimental work done on tokamak devices. The results in this work verified that the theory is still applicable to the RFP, in spite of its more complicated magnetic mode structure, involving perturbations of a single poloidal mode number

Analysis of resistive tearing-mode in the reversed-field pinch plasma

International Nuclear Information System (INIS)

Oshiyama, Hiroshi; Masamune, Sadao; Hamuro, Eitaro; Tamaki, Reiji.

1985-01-01

As one of the methods of confining high temperature plasma by magnetic stress, attention has been paid to reversed field pinch (RFP). This RFP is the method of maintaining plasma pressure by combining the poloidal field generated by plasma current and the toroidal field having nearly same intensity, thus forming the toroidal shape, closed magnetic surface. As the typical RFP equipment, there have been TPE-1R(M), HBTX-1A, ZT-40M and OHTE, but in order to anticipate the further development, one of the problems is the resistive instability. In this study, the critical beta value determined by the tearing mode in RFP configuration was examined by analytical and numerical calculation methods. The position of a wall required for the stability was determined by solving a second order differential equation for a radial perturbed magnetic field. The propriety of the computer code for determining the position was examined. The magnetic field configuration having a finite beta value was determined, and its stability against a tearing mode was investigated. For this judgement of the stability, the developed computer code was used. The tearing mode in a Bessel function model, the tearing mode of a finite beta value and others are described. (Kako, I.)

Power balance in an Ohmically heated fusion reactor

International Nuclear Information System (INIS)

Christiansen, J.P.; Roberts, K.V.

1982-01-01

A simplified power-balance equation (zero-dimensional model) is used to study the performance of an Ohmically heated fusion reactor with emphasis on a pulsed reversed-field pinch concept (RFP). The energy confinement time tausub(E) is treated as an adjustable function, and empirical tokamak scaling laws are employed in the numerical estimates, which are supplemented by 1-D ATHENE code calculations. The known heating rates and energy losses are represented by the net energy replacement time tausub(W), which is exhibited as a surface in density (n) and temperature (T) space with a saddle point (nsub(*), Tsub(*)), the optimum ignition point. It is concluded that i) ignition by Ohmic heating is more practicable for the RFP reactor than for a tokamak reactor with the same tausub(E), (ii) if at fixed current the minor radius can be reduced or at fixed minor radius the current can be increased, then it is found that Ohmic ignition becomes more likely when present tokamak scaling laws are used. More definitive estimates require, however, a knowledge of tausub(E), which can only be obtained by establishing a reliable set of experimental RFP scaling laws and, in particular, by extending RFP experiments closer to the reactor regime. (author)

Characteristics of the TPE reversed-field pinch plasmas in conventional and improved confinement regimes

International Nuclear Information System (INIS)

Sakakita, H.; Asai, T.; Fiksel, G.; Yagi, Y.; Frassinetti, L.; Hayase, K.; Hirano, Y.; Kiyama, S.; Koguchi, H.; Shimada, T.; Innocente, P.; Spizzo, G.; Terranova, D.; Sato, Y.; Yoshikawa, M.

2005-01-01

We present the characteristics and experimental scaling laws of reversed-field pinch (RFP) plasmas, which are obtained from the recently established toroidal pinch experiment (TPE) database. The database contains information for approximately 1500 discharges consistently selected from four TPE RFP devices, and covers two decades of RFP experiments under conventional operating conditions at the National Institute of Advanced Industrial Science and Technology. We present the physics of the pulsed poloidal current drive (PPCD) discharges in the TPE-RX RFP device, and a comparison of the improved energy confinement time in PPCD, τ E P PCD , with τ E s caling as the reference scaling law (τ E s caling ∼ a 1.63 (I P 0.78 (I P /N) 0.33 Θ 2.97 ) in the TPE database, is attempted. The result shows that τ E P PCD agrees well with τ E s caling because of the strong pinch parameter dependence on the TPE scaling law. A potential improved confinement mode in the quasi-single-helicity (QSH) state is also investigated in TPE-RX, with respect to the operation conditions under which the QSH spontaneously appears in the core region, where a typical island structure is observed by means of soft Xray tomography. (author)

Genetic Recombination Between Stromal and Cancer Cells Results in Highly Malignant Cells Identified by Color-Coded Imaging in a Mouse Lymphoma Model.

Science.gov (United States)

Nakamura, Miki; Suetsugu, Atsushi; Hasegawa, Kousuke; Matsumoto, Takuro; Aoki, Hitomi; Kunisada, Takahiro; Shimizu, Masahito; Saji, Shigetoyo; Moriwaki, Hisataka; Hoffman, Robert M

2017-12-01

The tumor microenvironment (TME) promotes tumor growth and metastasis. We previously established the color-coded EL4 lymphoma TME model with red fluorescent protein (RFP) expressing EL4 implanted in transgenic C57BL/6 green fluorescent protein (GFP) mice. Color-coded imaging of the lymphoma TME suggested an important role of stromal cells in lymphoma progression and metastasis. In the present study, we used color-coded imaging of RFP-lymphoma cells and GFP stromal cells to identify yellow-fluorescent genetically recombinant cells appearing only during metastasis. The EL4-RFP lymphoma cells were injected subcutaneously in C57BL/6-GFP transgenic mice and formed subcutaneous tumors 14 days after cell transplantation. The subcutaneous tumors were harvested and transplanted to the abdominal cavity of nude mice. Metastases to the liver, perigastric lymph node, ascites, bone marrow, and primary tumor were imaged. In addition to EL4-RFP cells and GFP-host cells, genetically recombinant yellow-fluorescent cells, were observed only in the ascites and bone marrow. These results indicate genetic exchange between the stromal and cancer cells. Possible mechanisms of genetic exchange are discussed as well as its ramifications for metastasis. J. Cell. Biochem. 118: 4216-4221, 2017. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

Monitoring protein turnover during phosphate starvation-dependent autophagic degradation using a photoconvertible fluorescent protein aggregate in tobacco BY-2 cells.

Science.gov (United States)

Tasaki, Maiko; Asatsuma, Satoru; Matsuoka, Ken

2014-01-01

We have developed a system for quantitative monitoring of autophagic degradation in transformed tobacco BY-2 cells using an aggregate-prone protein comprised of cytochrome b5 (Cyt b5) and a tetrameric red fluorescent protein (RFP). Unfortunately, this system is of limited use for monitoring the kinetics of autophagic degradation because the proteins synthesized before and after induction of autophagy cannot be distinguished. To overcome this problem, we developed a system using kikume green-red (KikGR), a photoconvertible and tetrameric fluorescent protein that changes its fluorescence from green to red upon irradiation with purple light. Using the fusion protein of Cyt b5 and KikGR together with a method for the bulk conversion of KikGR, which we had previously used to convert the Golgi-localized monomeric KikGR fusion protein, we were able to monitor both the growth and de novo formation of aggregates. Using this system, we found that tobacco cells do not cease protein synthesis under conditions of phosphate (Pi)-starvation. Induction of autophagy under Pi-starvation, but not under sugar- or nitrogen-starvation, was specifically inhibited by phosphite, which is an analog of Pi with a different oxidation number. Therefore, the mechanism by which BY-2 cells can sense Pi-starvation and induce autophagy does not involve sensing a general decrease in energy supply and a specific Pi sensor might be involved in the induction of autophagy under Pi-starvation.

Changes in risk factor profile after ischemic stroke

DEFF Research Database (Denmark)

Hornnes, Nete

up 1 year after stroke. We constructed a baseline risk factor profile (RFP) of 6 variables: smoking, excessive drinking, physical inactivity, untreated hypertension, no cholesterol-lowering, and no antithrombotic treatment/warfarin at discharge from hospital. Each item was rated 0 or 1 giving...... a maximum score of 6 points. Mean baseline RFP-score was 1.6 Results. After 1 year we found a reduction in current smoking (p=0.008) and in excessive drinking (p=0.0001). There was no change in physical activity and in untreated hypertension. There was an increase in the proportion of patients on lipid......-lowering (p=0.011) and antithrombotic (p=0.0003) treatment yielding a reduction in RFP to 1.4 (phypertensive, and most patients with untreated hypertension and hypercholesterolemia remained untreated. By 1-year follow up 30 patients (9.4%) had had a non...

Action of DTPA on hepatic plutonium. III. Evidence for a direct chelation mechanism for DTPA-induced excretion of monomeric plutonium into rat bile

International Nuclear Information System (INIS)

Bhattacharyya, M.H.; Peterson, D.P.

1979-01-01

Trisodium calcium diethylenetriaminepenta[2- 14 C]acetic acid ([ 14 C]DTPA, 0.25 nmole/kg, 0.210 mCi/mmole) was administered via the jugular vein to three rats whose bile ducts and urinary bladders were cannulated. Bile and urine were collected for 24 h after injection, and tissues and excreta were then analyzed for 14 C content. [ 14 C]DTPA was identified in the bile samples by cochromatography with unlabeled DTPA on an anion-exchange column system. Data from this experiment demonstrated the existence of a minor excretion pathway for DTPA into rat bile, accounting for 0.12% of the injected dose by 24 h after administration. Bile was collected for 24 h following the administration of unlabeled Na 3 [CaDTPA] (0.25 mmole/kg, iv) to a rat which had received 239 Pu(IV)-citrate (5.7 μCi/kg, iv) 24 h prior to DTPA. Bile samples containing Pu were analyzed using anion-exchange column chromatography. Eighty to ninety percent of the Pu present in bile chromatographed as a single peak in the position of the Pu-DTPA complex. Cochromatograhy of the 0- to 6-h bile sample with a 239 Pu-[ 14 C]DTPA complex revealed that the Pu appearing in bile following DTPA treatment comigrated with the 239 Pu-[ 14 C]DTPA complex. These data provide strong evidence that, in the rat, DTPA acts to remove hepatic deposits of monomeric Pu by formation of the Pu-DTPA complex followed by excretion of the complex into bile

Fascinating transformations of donor-acceptor complexes of group 13 metal (Al, Ga, In) derivatives with nitriles and isonitriles: from monomeric cyanides to rings and cages.

Science.gov (United States)

Timoshkin, Alexey Y; Schaefer, Henry F

2003-08-20

Formation of the donor-acceptor complexes of group 13 metal derivatives with nitriles and isonitriles X(3)M-D (M = Al,Ga,In; X = H,Cl,CH(3); D = RCN, RNC; R = H,CH(3)) and their subsequent reactions have been theoretically studied at the B3LYP/pVDZ level of theory. Although complexation with MX(3) stabilizes the isocyanide due to the stronger M-C donor-acceptor bond, this stabilization (20 kJ mol(-1) at most) is not sufficient to make the isocyanide form more favorable. Relationships between the dissociation enthalpy DeltaH degrees (298)(diss), charge-transfer q(CT), donor-acceptor bond energy E(DA), and the shift of the vibrational stretching mode of the CN group upon coordination Deltaomega(CN) have been examined. For a given metal center, there is a good correlation between the energy of the donor-acceptor bond and the degree of a charge transfer. Prediction of the DeltaH degrees (298)(diss) on the basis of the shift of CN stretching mode is possible within limited series of cyanide complexes (for the fixed M,R); in contrast, complexes of the isocyanides exhibit very poor Deltaomega(CN) - DeltaH degrees (298)(diss) correlation. Subsequent X ligand transfer and RX elimination reactions yielding monomeric (including donor-acceptor stabilized) and variety of oligomeric cage and ring compounds with [MN]n, [MC]n, [MNC]n cores have been considered and corresponding to thermodynamic characteristics have been obtained for the first time. Monomeric aluminum isocyanides X(2)AlNC are more stable compared to Al-C bonded isomers; for gallium and indium situation is reversed, in qualitative agreement with Pearson's HSAB concept. Substitution of X by CN in MX(3) increases the dissociation enthalpy of the MX(2)CN-NH(3) complex compared to that for MX(3)-NH(3), irrespective of the substituent X. Mechanisms of the initial reaction of the X transfer have been studied for the case X = R = H. The process of hydrogen transfer from the metal to the carbon atom in H(3)M-CNH is

RFP for the italien satellite AGILE

DEFF Research Database (Denmark)

Madsen, Peter Buch; Jørgensen, John Leif; Riis, Troels

1999-01-01

The document descibes the ASC Star Tracker (performance, functionality, requirements etc.) to the Italian satellite AGILE.......The document descibes the ASC Star Tracker (performance, functionality, requirements etc.) to the Italian satellite AGILE....

Microinstabilities and turbulent transport in the reversed field pinch

Science.gov (United States)

Carmody, Daniel Richard

The work presented in this thesis is concerned with addressing the nature of drift wave microturbulence in the reversed field pinch (RFP). Microturbulence is an important phenomenon and contributor to heat and particle transport in tokamaks, where it has been studied for several decades, but its role in the RFP is a rather new topic of study. As such, the nature of RFP drift waves and their relationship to their tokamak counterparts is still developing, and many of the results in this work are focused on addressing this challenge. Fundamental advances in microturbulence research have been made in recent decades through two parallel developments: the theoretical framework encompassed in the gyrokinetic model, and the computational power offered by massively-parallel, high-performance computing systems. Gyrokinetics is a formulation of kinetic theory in such a way that the fast timescale gyromotion of particles around magnetic field lines is averaged out. The implementation and use of RFP equilibrium models in gyrokinetic codes constitutes the bulk of this thesis. A simplified analytic equilibrium, the toroidal Bessel function model (TBFM), is used in the gyrokinetic code GYRO to explore the fundamental scaling properties of drift waves in the RFP geometry. Two drift wave instabilities, the ion temperature gradient (ITG) mode and the microtearing mode (MTM) are found to occur, and the relationship of their critical threshold in driving gradients and plasma beta is explored. The critical values in these parameters are found to be above those of similar tokamak cases by roughly a factor of the flux surface aspect ratio. The MTM is found to be stabilized by increasing the RFP pinch parameter theta, making it unlikely for it to unstable in the high-theta improved confinement pulsed poloidal current drive (PPCD) discharges. Efforts are also made to address microinstabilities in specific experimental discharges of the Madison Symmetric Torus (MST). A semi

Preparation and characterization of Schiff base Cu(II) complex and its applications on textile materials

Science.gov (United States)

Oylumluoglu, G.; Oner, J.

2017-10-01

Schiff base ligands are regarded as an important class of organic compounds on account of the fact that their complexation ability with transition metal ions. A new monomeric Schiff base Cu(II) complex, [Cu(HL)2], 1 [H2L = 2-((E)-(2-hydroxypropylimino)methyl)-4-nitrophenol] has been synthesized and characterized by elemental analysis, UV and IR spectroscopy, single crystal X-ray diffraction and photoluminescence study. While the Schiff base ligand and its Cu(II) complex are excited at λex = 349 nm in UV region, the Schiff base ligand shows a blue emission band at λmax = 480 nm whereas its Cu(II) complex shows a strong green emission band at λmax = 520 nm in the solid state at room temperature. The luminescent properties showed that the Schiff base ligand and its Cu(II) complex can be used as novel potential candidates for applications in textile such as UV-protection, antimicrobial, laundry and functional bleaching treatments.

Modulation of Amyloid-β Conformation by Charge State of N-Terminal Disordered Region

International Nuclear Information System (INIS)

Xi Wen-Hui; Li Wen-Fei; Wang Wei

2012-01-01

Based on molecular dynamics simulations, we show that variations of the charge states of the histidines, which are the main effects of pH-value change and metal binding, can lead to a drastic change of the intra-peptide interactions of the segment 17–42 and the conformational distribution of the monomeric amyloid-β (Aβ). Since we already knew that the conformational distribution of monomeric Aβ can largely affect Aβ fibrillar aggregation, our results suggest that the pH value change and metal binding can affect the Aβ aggregation by much more complex mechanism than just affecting the inter-peptide interactions. To fully understand the mechanism of metal binding and pH-value induced Aβ aggregation, we also need to consider their effects on the conformational distribution of monomeric Aβ. (cross-disciplinary physics and related areas of science and technology)

Prospects for fusion applications of reversed-field pinches

International Nuclear Information System (INIS)

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

1985-01-01

The applicability of the Reversed-Field Pinch (RFP) as a source of fusion neutrons for use in developing key fusion nuclear technologies is examined. This Fusion Test Facility (FTF) would emphasize high neutron wall loading, small plasma volume, low fusion and driver powers, and steady-state operation. Both parametric tradeoffs based on present-day physics understanding and a conceptual design based on an approx.1-MW/m 2 (neutron) driven operation are reported. 10 refs

Functional imaging in bulk tissue specimens using optical emission tomography: fluorescence preservation during optical clearing

International Nuclear Information System (INIS)

Sakhalkar, H S; Dewhirst, M; Oliver, T; Cao, Y; Oldham, M

2007-01-01

Optical emission computed tomography (optical-ECT) is a technique for imaging the three-dimensional (3D) distribution of fluorescent probes in biological tissue specimens with high contrast and spatial resolution. In optical-ECT, functional information can be imaged by (i) systemic application of functional labels (e.g. fluorophore labelled proteins) and/or (ii) endogenous expression of fluorescent reporter proteins (e.g. red fluorescent protein (RFP), green fluorescent protein (GFP)) in vivo. An essential prerequisite for optical-ECT is optical clearing, a procedure where tissue specimens are made transparent to light by sequential perfusion with fixing, dehydrating and clearing agents. In this study, we investigate clearing protocols involving a selection of common fixing (4% buffered paraformaldehyde (PFA), methanol and ethanol), dehydrating (methanol and ethanol) and clearing agents (methyl salicylate and benzyl-alcohol-benzyl-benzoate (BABB)) in order to determine a 'fluorescence friendly' clearing procedure. Cell culture experiments were employed to optimize the sequence of chemical treatments that best preserve fluorescence. Texas red (TxRed), fluorescein isothiocyanate (FITC), RFP and GFP were tested as fluorophores and fluorescent reporter proteins of interest. Fluorescent and control cells were imaged on a microscope using a DSred2 and FITC filter set. The most promising clearing protocols of cell culture experiments were applied to whole xenograft tumour specimens, to test their effectiveness in large unsectioned samples. Fluorescence of TxRed/FITC fluorophores was not found to be significantly affected by any of the test clearing protocols. RFP and GFP fluorescence, however, was found to be significantly greater when cell fixation was in ethanol. Fixation in either PFA or methanol resulted in diminished fluorescence. After ethanol fixation, the RFP and GFP fluorescence proved remarkably robust to subsequent exposure to either methyl salicylate or BABB

Functional imaging in bulk tissue specimens using optical emission tomography: fluorescence preservation during optical clearing

Energy Technology Data Exchange (ETDEWEB)

Sakhalkar, H S [Department of Radiation Oncology, Duke University Medical Center, Durham, NC 27710 (United States); Dewhirst, M [Department of Radiation Oncology, Duke University Medical Center, Durham, NC 27710 (United States); Oliver, T [Department of Cell Biology, Duke University Medical Center, Durham, NC 27710 (United States); Cao, Y [Department of Radiation Oncology, Duke University Medical Center, Durham, NC 27710 (United States); Oldham, M [Department of Radiation Oncology Physics, and Biomedical Engineering, Duke University Medical Center, Durham, NC 27710 (United States)

2007-04-21

Optical emission computed tomography (optical-ECT) is a technique for imaging the three-dimensional (3D) distribution of fluorescent probes in biological tissue specimens with high contrast and spatial resolution. In optical-ECT, functional information can be imaged by (i) systemic application of functional labels (e.g. fluorophore labelled proteins) and/or (ii) endogenous expression of fluorescent reporter proteins (e.g. red fluorescent protein (RFP), green fluorescent protein (GFP)) in vivo. An essential prerequisite for optical-ECT is optical clearing, a procedure where tissue specimens are made transparent to light by sequential perfusion with fixing, dehydrating and clearing agents. In this study, we investigate clearing protocols involving a selection of common fixing (4% buffered paraformaldehyde (PFA), methanol and ethanol), dehydrating (methanol and ethanol) and clearing agents (methyl salicylate and benzyl-alcohol-benzyl-benzoate (BABB)) in order to determine a 'fluorescence friendly' clearing procedure. Cell culture experiments were employed to optimize the sequence of chemical treatments that best preserve fluorescence. Texas red (TxRed), fluorescein isothiocyanate (FITC), RFP and GFP were tested as fluorophores and fluorescent reporter proteins of interest. Fluorescent and control cells were imaged on a microscope using a DSred2 and FITC filter set. The most promising clearing protocols of cell culture experiments were applied to whole xenograft tumour specimens, to test their effectiveness in large unsectioned samples. Fluorescence of TxRed/FITC fluorophores was not found to be significantly affected by any of the test clearing protocols. RFP and GFP fluorescence, however, was found to be significantly greater when cell fixation was in ethanol. Fixation in either PFA or methanol resulted in diminished fluorescence. After ethanol fixation, the RFP and GFP fluorescence proved remarkably robust to subsequent exposure to either methyl salicylate

The helical MreB cytoskeleton in Escherichia coli MC1000/pLE7 is an artifact of the N-Terminal yellow fluorescent protein tag.

Science.gov (United States)

Swulius, Matthew T; Jensen, Grant J

2012-12-01

Based on fluorescence microscopy, the actin homolog MreB has been thought to form extended helices surrounding the cytoplasm of rod-shaped bacterial cells. The presence of these and other putative helices has come to dominate models of bacterial cell shape regulation, chromosome segregation, polarity, and motility. Here we use electron cryotomography to show that MreB does in fact form extended helices and filaments in Escherichia coli when yellow fluorescent protein (YFP) is fused to its N terminus but native (untagged) MreB expressed to the same levels does not. In contrast, mCherry fused to an internal loop (MreB-RFP(SW)) does not induce helices. The helices are therefore an artifact of the placement of the fluorescent protein tag. YFP-MreB helices were also clearly distinguishable from the punctate, "patchy" localization patterns of MreB-RFP(SW), even by standard light microscopy. The many interpretations in the literature of such punctate patterns as helices should therefore be reconsidered.

Studies of a poloidal divertor reversed field pinch

International Nuclear Information System (INIS)

Sarff, J.S.; Almagri, A.F.; Assadi, S.; Den Hartog, D.J.; Dexter, R.N.; Prager, S.C.; Sprott, J.C.

1988-07-01

An attempt has been made to form a reversed field pinch (RFP) in a poloidal divertor configuration which position the plasma far from a conducting wall. In this configuration, the plasma is localized within a magnetic separatrix formed by the combination of toroidal currents in the plasma and four internal aluminum rings. Plasmas were formed with plasma current /approximately/135 kA, toroidal field reversal lasting /approximately/1 msec, line-averaged density /approximately/1--2 /times/ 10 13 cm/sup /minus/3/ and central electron temperature /approximately/55 eV, but a large asymmetry in the magnetic field (δB/B /approximately/40%) onset at about the time the toroidal field reversed at the wall. Symmetric, poloidal divertor RFP equilibria were not formed. This behavior might be expected based on linear MHD stability analysis of a cylindrical plasma bounded by a large vacuum region and distant conducting wall. The symmetric equilibrium before the asymmetry develops and the asymmetry itself are described. 15 refs., 3 figs

The effect of collisionality and diamagnetism on the plasma dynamo

International Nuclear Information System (INIS)

Ji, H.; Yagi, Y.; Hattori, K.; Hirano, Y.; Shimada, T.; Maejima, Y.; Hayase, K.; Almagri, A.F.; Prager, S.C.; Sarff, J.S.

1995-01-01

Fluctuation-induced dynamo forces are measured over a wide range of electron collisionality in the edge of TPE-1RM20 Reversed-Field Pinch (RFP). In the collisionless region the Magnetohydrodynamic (MHD) dynamo alone can sustain the parallel current, while in the collisional region a new dynamo mechanism resulting from the fluctuations in the electron diamagnetic drift becomes dominant. A comprehensive picture of the RFP dynamo emerges by combining with earlier results from MST and REPUTE RFPs

Electrical circuit modeling of reversed field pinches

International Nuclear Information System (INIS)

Sprott, J.C.

1988-02-01

Equations are proposed to describe the radial variation of the magnetic field and current density in a circular, cylindrical RFP. These equations are used to derive the electrical circuit parameters (inductance, resistance, and coupling coefficient) for an RFP discharge. The circuit parameters are used to evaluate the flux and energy consumption for various startup modes and for steady-state operation using oscillating field current drive. The results are applied to the MST device. 32 refs., 14 figs., 1 tab

RCRA closures at Rocky Flats Plant: A programmatic perspective and case study

International Nuclear Information System (INIS)

Ogg, Randy T.; Peterman, Bruce D.

1992-01-01

The Interagency Agreement (IAG) integrates a unique mechanism for remediating hazardous waste sites at the Rocky Flats Plant (RFP), which include utilizing RCRA and CERCLA technical/regulatory processes. Pursuant to the IAG signed by the Department of Energy (DOE), Environmental Protection Agency (EPA), and the Colorado Department of Health (CDH) on January 22, 1991, sixteen operable units (OUs) were defined for characterization and remediation at RFP. Of the sixteen OUs, six are classified as Resource Conservation and Recovery Act (RCRA) closure units. The six RCRA interim status closure units are: Solar Evaporation Ponds-OU 4, Present LandfUl-OU 7, Original Process Waste Lines-OU 9, Other Outside Closures-OU 10, West Spray Field-OU II, and Inside Building Closures-OU 15. The IAG will function as a technical/regulatory mechanism for managing/complying with all aspects of the RCRA interim status closure units at RFP. (author)

Transport simulations of the oscillating field current drive experiment in the ZT-40M reversed field pinch

International Nuclear Information System (INIS)

Scardovelli, R.A.; Nebel, R.A.; Werley, K.A.; Miley, G.H.

1987-01-01

Oscillating Field Current Drive (OFCD) is based on the premise that in order to sustain a relaxing Reversed Field Pinch (RFP) plasma, one needs only to supply magnetic helicity at the same rate it is consumed. The purpose of this work is to try to better understand the possible mechanisms underlying these relaxations within the context of different kinds of resistive MHD instabilities

Contribution of Underlying Connective Tissue Cells to Taste Buds in Mouse Tongue and Soft Palate

Science.gov (United States)

Mederacke, Ingmar; Komatsu, Yoshihiro; Stice, Steve; Schwabe, Robert F.; Mistretta, Charlotte M.; Mishina, Yuji; Liu, Hong-Xiang

2016-01-01

Taste buds, the sensory organs for taste, have been described as arising solely from the surrounding epithelium, which is in distinction from other sensory receptors that are known to originate from neural precursors, i.e., neural ectoderm that includes neural crest (NC). Our previous study suggested a potential contribution of NC derived cells to early immature fungiform taste buds in late embryonic (E18.5) and young postnatal (P1-10) mice. In the present study we demonstrated the contribution of the underlying connective tissue (CT) to mature taste buds in mouse tongue and soft palate. Three independent mouse models were used for fate mapping of NC and NC derived connective tissue cells: (1) P0-Cre/R26-tdTomato (RFP) to label NC, NC derived Schwann cells and derivatives; (2) Dermo1-Cre/RFP to label mesenchymal cells and derivatives; and (3) Vimentin-CreER/mGFP to label Vimentin-expressing CT cells and derivatives upon tamoxifen treatment. Both P0-Cre/RFP and Dermo1-Cre/RFP labeled cells were abundant in mature taste buds in lingual taste papillae and soft palate, but not in the surrounding epithelial cells. Concurrently, labeled cells were extensively distributed in the underlying CT. RFP signals were seen in the majority of taste buds and all three types (I, II, III) of differentiated taste bud cells, with the neuronal-like type III cells labeled at a greater proportion. Further, Vimentin-CreER labeled cells were found in the taste buds of 3-month-old mice whereas Vimentin immunoreactivity was only seen in the CT. Taken together, our data demonstrate a previously unrecognized origin of taste bud cells from the underlying CT, a conceptually new finding in our knowledge of taste bud cell derivation, i.e., from both the surrounding epithelium and the underlying CT that is primarily derived from NC. PMID:26741369

Contribution of Underlying Connective Tissue Cells to Taste Buds in Mouse Tongue and Soft Palate.

Directory of Open Access Journals (Sweden)

Kristin Boggs

Full Text Available Taste buds, the sensory organs for taste, have been described as arising solely from the surrounding epithelium, which is in distinction from other sensory receptors that are known to originate from neural precursors, i.e., neural ectoderm that includes neural crest (NC. Our previous study suggested a potential contribution of NC derived cells to early immature fungiform taste buds in late embryonic (E18.5 and young postnatal (P1-10 mice. In the present study we demonstrated the contribution of the underlying connective tissue (CT to mature taste buds in mouse tongue and soft palate. Three independent mouse models were used for fate mapping of NC and NC derived connective tissue cells: (1 P0-Cre/R26-tdTomato (RFP to label NC, NC derived Schwann cells and derivatives; (2 Dermo1-Cre/RFP to label mesenchymal cells and derivatives; and (3 Vimentin-CreER/mGFP to label Vimentin-expressing CT cells and derivatives upon tamoxifen treatment. Both P0-Cre/RFP and Dermo1-Cre/RFP labeled cells were abundant in mature taste buds in lingual taste papillae and soft palate, but not in the surrounding epithelial cells. Concurrently, labeled cells were extensively distributed in the underlying CT. RFP signals were seen in the majority of taste buds and all three types (I, II, III of differentiated taste bud cells, with the neuronal-like type III cells labeled at a greater proportion. Further, Vimentin-CreER labeled cells were found in the taste buds of 3-month-old mice whereas Vimentin immunoreactivity was only seen in the CT. Taken together, our data demonstrate a previously unrecognized origin of taste bud cells from the underlying CT, a conceptually new finding in our knowledge of taste bud cell derivation, i.e., from both the surrounding epithelium and the underlying CT that is primarily derived from NC.

Using Dual Fluorescence Reporting Genes to Establish an In Vivo Imaging Model of Orthotopic Lung Adenocarcinoma in Mice.

Science.gov (United States)

Lai, Cheng-Wei; Chen, Hsiao-Ling; Yen, Chih-Ching; Wang, Jiun-Long; Yang, Shang-Hsun; Chen, Chuan-Mu

2016-12-01

Lung adenocarcinoma is characterized by a poor prognosis and high mortality worldwide. In this study, we purposed to use the live imaging techniques and a reporter gene that generates highly penetrative near-infrared (NIR) fluorescence to establish a preclinical animal model that allows in vivo monitoring of lung cancer development and provides a non-invasive tool for the research on lung cancer pathogenesis and therapeutic efficacy. A human lung adenocarcinoma cell line (A549), which stably expressed the dual fluorescence reporting gene (pCAG-iRFP-2A-Venus), was used to generate subcutaneous or orthotopic lung cancer in nude mice. Cancer development was evaluated by live imaging via the NIR fluorescent signals from iRFP, and the signals were verified ex vivo by the green fluorescence of Venus from the gross lung. The tumor-bearing mice received miR-16 nucleic acid therapy by intranasal administration to demonstrate therapeutic efficacy in this live imaging system. For the subcutaneous xenografts, the detection of iRFP fluorescent signals revealed delicate changes occurring during tumor growth that are not distinguishable by conventional methods of tumor measurement. For the orthotopic xenografts, the positive correlation between the in vivo iRFP signal from mice chests and the ex vivo green fluorescent signal from gross lung tumors and the results of the suppressed tumorigenesis by miR-16 treatment indicated that lung tumor size can be accurately quantified by the emission of NIR fluorescence. In addition, orthotopic lung tumor localization can be accurately visualized using iRFP fluorescence tomography in vivo, thus revealing the trafficking of lung tumor cells. We introduced a novel dual fluorescence lung cancer model that provides a non-invasive option for preclinical research via the use of NIR fluorescence in live imaging of lung.

Preparation, characterization, and in vitro cytotoxicity evaluation of a novel anti-tuberculosis reconstruction implant.

Directory of Open Access Journals (Sweden)

JunFeng Dong

Full Text Available Reconstruction materials currently used in clinical for osteoarticular tuberculosis (TB are unsatisfactory due to a variety of reasons. Rifampicin (RFP is a well-known and highly effective first-line anti-tuberculosis (anti-TB drug. Poly-DL-lactide (PDLLA and nano-hydroxyapatite (nHA are two promising materials that have been used both for orthopedic reconstruction and as carriers for drug release. In this study we report the development of a novel anti-TB implant for osteoarticular TB reconstruction using a combination of RFP, PDLLA and nHA.RFP, PDLLA and nHA were used as starting materials to produce a novel anti-TB activity implant by the solvent evaporation method. After manufacture, the implant was characterized and its biodegradation and drug release profile were tested. The in vitro cytotoxicity of the implant was also evaluated in pre-osteoblast MC3T3-E1 cells using multiple methodologies.A RFP/PDLLA/nHA composite was successfully synthesized using the solvent evaporation method. The composite has a loose and porous structure with evenly distributed pores. The production process was steady and no chemical reaction occurred as proved by Fourier Transform Infrared Spectroscopy (FTIR and X-Ray Diffraction (XRD. Meanwhile, the composite blocks degraded and released drug for at least 12 weeks. Evaluation of in vitro cytotoxicity in MC3T3-E1 cells verified that the synthesized composite blocks did not affect cell growth and proliferation.It is feasible to manufacture a novel bioactive anti-TB RFP/PDLLA/nHA composite by the solvent evaporation method. The composite blocks showed appropriate properties such as degradation, drug release and biosafety to MC3T3-E1 cells. In conclusion, the novel composite blocks may have great potential for clinical applications in repairing bone defects caused by osteoarticular TB.

Contribution of Underlying Connective Tissue Cells to Taste Buds in Mouse Tongue and Soft Palate.

Science.gov (United States)

Boggs, Kristin; Venkatesan, Nandakumar; Mederacke, Ingmar; Komatsu, Yoshihiro; Stice, Steve; Schwabe, Robert F; Mistretta, Charlotte M; Mishina, Yuji; Liu, Hong-Xiang

2016-01-01

Taste buds, the sensory organs for taste, have been described as arising solely from the surrounding epithelium, which is in distinction from other sensory receptors that are known to originate from neural precursors, i.e., neural ectoderm that includes neural crest (NC). Our previous study suggested a potential contribution of NC derived cells to early immature fungiform taste buds in late embryonic (E18.5) and young postnatal (P1-10) mice. In the present study we demonstrated the contribution of the underlying connective tissue (CT) to mature taste buds in mouse tongue and soft palate. Three independent mouse models were used for fate mapping of NC and NC derived connective tissue cells: (1) P0-Cre/R26-tdTomato (RFP) to label NC, NC derived Schwann cells and derivatives; (2) Dermo1-Cre/RFP to label mesenchymal cells and derivatives; and (3) Vimentin-CreER/mGFP to label Vimentin-expressing CT cells and derivatives upon tamoxifen treatment. Both P0-Cre/RFP and Dermo1-Cre/RFP labeled cells were abundant in mature taste buds in lingual taste papillae and soft palate, but not in the surrounding epithelial cells. Concurrently, labeled cells were extensively distributed in the underlying CT. RFP signals were seen in the majority of taste buds and all three types (I, II, III) of differentiated taste bud cells, with the neuronal-like type III cells labeled at a greater proportion. Further, Vimentin-CreER labeled cells were found in the taste buds of 3-month-old mice whereas Vimentin immunoreactivity was only seen in the CT. Taken together, our data demonstrate a previously unrecognized origin of taste bud cells from the underlying CT, a conceptually new finding in our knowledge of taste bud cell derivation, i.e., from both the surrounding epithelium and the underlying CT that is primarily derived from NC.

Development of an image-based screening system for inhibitors of the plastidial MEP pathway and of protein geranylgeranylation

Science.gov (United States)

Hartmann, Michael; Gas-Pascual, Elisabet; Hemmerlin, Andrea; Rohmer, Michel; Bach, Thomas J.

2015-01-01

In a preceding study we have recently established an in vivo visualization system for the geranylgeranylation of proteins in a stably transformed tobacco BY-2 cell line, which involves expressing a dexamethasone-inducible GFP fused to the prenylable, carboxy-terminal basic domain of the rice calmodulin CaM61, which naturally bears a CaaL geranylgeranylation motif (GFP-BD-CVIL). By using pathway-specific inhibitors it was there demonstrated that inhibition of the methylerythritol phosphate (MEP) pathway with oxoclomazone and fosmidomycin, as well as inhibition of protein geranylgeranyl transferase type 1 (PGGT-1), shifted the localization of the GFP-BD-CVIL protein from the membrane to the nucleus. In contrast, the inhibition of the mevalonate (MVA) pathway with mevinolin did not affect this localization. Furthermore, in this initial study complementation assays with pathway-specific intermediates confirmed that the precursors for the cytosolic isoprenylation of this fusion protein are predominantly provided by the MEP pathway. In order to optimize this visualization system from a more qualitative assay to a statistically trustable medium or a high-throughput screening system, we established now new conditions that permit culture and analysis in 96-well microtiter plates, followed by fluorescence microscopy. For further refinement, the existing GFP-BD-CVIL cell line was transformed with an estradiol-inducible vector driving the expression of a RFP protein, C-terminally fused to a nuclear localization signal (NLS-RFP). We are thus able to quantify the total number of viable cells versus the number of inhibited cells after various treatments. This approach also includes a semi-automatic counting system, based on the freely available image processing software. As a result, the time of image analysis as well as the risk of user-generated bias is reduced to a minimum. Moreover, there is no cross-induction of gene expression by dexamethasone and estradiol, which is an

Design and manufacturing of vacuum vessel of TPE-RX

Energy Technology Data Exchange (ETDEWEB)

Sago, H.; Kaguchi, H.; Orita, J.; Ishigami, Y. [Mitsubishi Heavy Industries Ltd., Kobe (Japan); Urata, K. [Mitsubishi Heavy Industries Ltd. (Japan). Nuclear Energy Systems Engineering Center; Hasegawa, M. [Mitsubishi Electric Co. (Japan). Nuclear Fusion Development; Yagi, Y.; Hirano, Y.; Shimada, T.; Sekine, S.; Sakakita, H. [Electrotechnical Lab. (Japan)

1998-07-01

Construction of a new, large reversed field pinch (RFP) machine called TPE-RX was complete at the end of 1997 as a successor of the previous TPE-1RM20 machine at the Electrotechnical Laboratory (ETL). RFP configuration has been successfully obtained in March 1998. This paper introduces structural design and manufacturing of the vacuum vessel of TPE-RX. The support positions were decided by structural analyses. The structural integrity of the vacuum vessel was evaluated by inelastic analyses. (author)

Design and manufacturing of vacuum vessel of TPE-RX

International Nuclear Information System (INIS)

Sago, H.; Kaguchi, H.; Orita, J.; Ishigami, Y.; Urata, K.

1998-01-01

Construction of a new, large reversed field pinch (RFP) machine called TPE-RX was complete at the end of 1997 as a successor of the previous TPE-1RM20 machine at the Electrotechnical Laboratory (ETL). RFP configuration has been successfully obtained in March 1998. This paper introduces structural design and manufacturing of the vacuum vessel of TPE-RX. The support positions were decided by structural analyses. The structural integrity of the vacuum vessel was evaluated by inelastic analyses. (author)

Demonstration, testing and evaluation of nonintrusive characterization technologies at operable Unit 2 of Rocky Flats Plant. Final report

International Nuclear Information System (INIS)

1994-09-01

A three-dimensional (3-D), high-resolution (HR) seismic reflection evaluation was conducted at the Rocky Flats Plant (RFP), near Golden, Colorado, to demonstrate the applicability of nonintrusive characterization techniques to detect buried objects, contamination, and geological/hydrological features at RFP. The evaluation was conducted as part of the U.S. Department of Energy's (DOE) request for demonstration, testing and evaluation (DT ampersand E) of nonintrusive techniques, under DOE Program Research and Development Announcement (PRDA) No. DE-RA05-09OR22000

Demonstration, testing and evaluation of nonintrusive characterization technologies at operable Unit 2 of Rocky Flats Plant. Final report

Energy Technology Data Exchange (ETDEWEB)

NONE

1994-09-01

A three-dimensional (3-D), high-resolution (HR) seismic reflection evaluation was conducted at the Rocky Flats Plant (RFP), near Golden, Colorado, to demonstrate the applicability of nonintrusive characterization techniques to detect buried objects, contamination, and geological/hydrological features at RFP. The evaluation was conducted as part of the U.S. Department of Energy`s (DOE) request for demonstration, testing and evaluation (DT&E) of nonintrusive techniques, under DOE Program Research and Development Announcement (PRDA) No. DE-RA05-09OR22000.

Young children's reading for pleasure with digital books: six key facets of engagement

OpenAIRE

Kucirkova, N.; Littleton, K.; Cremin, T.

2017-01-01

This paper offers a new characterisation of young children’s (2–8 years) reading for pleasure (RfP) with digital books. This characterisation is rooted in a re-contextualisation of Anna Craft’s conceptualisation of twenty-first century childhoods in Creativity and education futures (Stoke on Trent, Trentham, 2011) and a review of the literature concerning young children’s RfP with digital books. The paper develops Craft’s (2011) work by considering the ways in which digital books can resource...

High beta plasma confinement and neoclassical effects in a small aspect ratio reversed field pinch

International Nuclear Information System (INIS)

Hayase, K.; Sugimoto, H.; Ashida, H.

2003-01-01

The high β equilibrium and stability of a reversed field pinch (RFP) configuration with a small aspect ratio are theoretically studied. The equilibrium profile, high beta limit and the bootstrap current effect on those are calculated. The Mercier stable critical β decreases with 1/A, but β∼0.2 is permissible at A=2 with help of edge current profile modification. The effect of bootstrap current is evaluated for various pressure and current profiles and cross-sectional shapes of plasma by a self-consistent neoclassical PRSM equilibrium formulation. The high bootstrap current fraction (F bs ) increases the shear stabilization effect in the core region, which enhances significantly the stability β limit compared with that for the classical equilibrium. These features of small aspect ratio RFP, high β and high F bs , and a possibly easier access to the quasi-single helicity state beside the intrinsic compact structure are attractive for the feasible economical RFP reactor concept. (author)

Approach to high stability beta limit and its control by fast wave current drive in reversed field pinch plasma

International Nuclear Information System (INIS)

Kusano, K.; Kondoh, Y.; Gesso, H.; Osanai, Y.; Saito, K.N.; Ukai, R.; Nanba, T.; Nagamine, Y.; Shiina, S.

2001-01-01

Before the generation of steady state, dynamo-free RFP configuration by rf current driving scheme, it is necessary to find an optimum configuration into high stability beta limit against m=1 resonant resistive MHD modes and reducing nonlinearly turbulent level with less rf power. As first step to the optimization study, we are interested in partially relaxed state model (PRSM) RFP configuration, which is considered to be closer to a relaxed state at finite beta since it has force-free fields for poloidal direction with a relatively shorter characteristic length of relaxation and a relatively higher stability beta limit to m=1 resonant ideal MHD modes. The stability beta limit to m=1 resonant resistive MHD modes can be predicted to be relatively high among other RFP models and to be enhanced by the current density profile control using fast magnetosonic waves (FMW), which are accessible to high density region with strong absorption rate. (author)

Comprehensive evaluation of future site alternatives

International Nuclear Information System (INIS)

Shain, D.I.; Jones, M.E.; Ryan, K.

1994-01-01

Due to rapid changes occurring within the Nuclear Weapons complex, the need for integrated planning designed to combine multiple program needs into one strategic plan has become a necessity. This is more apparent as diverse DOE programs compete for dwindling resources. These programs range from traditional production operations, environmental and waste management, to facility transition, economic development, decontamination and decommissioning, and environmental restoration activities. Each program can influence another, thus increasing the difficulty of distinguishing program elements. The method in developing comprehensive plans becomes even more complicated when environmental compliance issues, regulatory agreements and stakeholder values are considered. AT the Department of Energy's (DOE) Rocky Flats Plant (RFP), all of these program conditions exist. This paper addresses a set of tools which are being developed at RFP that provides key planning elements and alternatives assessment for the DOE's Office of Planning and Integration (OPI) and National Environmental Policy Act (NEPA) Compliance Officer at RFP. This set of tools is referred to as the Systems Engineering Analysis (SEA)

Confinement improvement with rf poloidal current drive in the reversed-field pinch

International Nuclear Information System (INIS)

Hokin, S.; Sarff, J.; Sovinec, C.; Uchimoto, E.

1994-01-01

External control of the current profile in a reversed-field pinch (RFP), by means such as rf poloidal current drive, may have beneficial effects well beyond the direct reduction of Ohmic input power due to auxiliary heating. Reduction of magnetic turbulence associated with the dynamo, which drives poloidal current in a conventional RFP, may allow operation at lower density and higher electron temperature, for which rf current drive becomes efficient and the RFP operates in a more favorable regime on the nτ vs T diagram. Projected parameters for RFX at 2 MA axe studied as a concrete example. If rf current drive allows RFX to operate with β = 10% (plasma energy/magnetic energy) at low density (3 x 10 19 m -3 ) with classical resistivity (i.e. without dynamo-enhanced power input), 40 ms energy confinement times and 3 keV temperatures will result, matching the performance of tokamaks of similar size

Electron transport in the stochastic fields of the reversed-field pinch

International Nuclear Information System (INIS)

Kim, M.-H.; Punjabi, A.

1996-01-01

We employ the Monte Carlo method for the calculation of anomalous transport developed by Punjabi and Boozer to calculate the particle diffusion coefficient for electrons in the stochastic magnetic fields of the reversed-field pinch (RFP). In the Monte Carlo calculations represented here, the transport mechanism is the loss of magnetic surfaces due to resistive perturbations. The equilibrium magnetic fields are represented by the Bessel function model for the RFP. The diffusion coefficient D is calculated as a function of a, the amplitude of the perturbation. We see three regimes as the amplitude of the tearing modes is increased: the Rechester-Rosenbluth regime where D scales as a 2 ; the anomalous regime where D scales more rapidly than a 2 ; and the Mynick-Krommes regime where D scales more slowly than a 2 . Inclusion of the effects of loop voltage on the particle drift orbits in the RFP does not affect the intervals in the amplitude a where these regimes operate. (Author)

Analysis methods for fast impurity ion dynamics data

International Nuclear Information System (INIS)

Den Hartog, D.J.; Almagri, A.F.; Prager, S.C.; Fonck, R.J.

1994-08-01

A high resolution spectrometer has been developed and used on the MST reversed-field pinch (RFP) to measure passively impurity ion temperatures and flow velocities with 10 μs temporal resolution. Such measurements of MHD-scale fluctuations are particularly relevant in the RFP because the flow velocity fluctuation induced transport of current (the ''MHD dynamo'') may produce the magnetic field reversal characteristic of an RFP. This instrument will also be used to measure rapid changes in the equilibrium flow velocity, such as occur during locking and H-mode transition. The precision of measurements made to date is <0.6 km/s. The authors are developing accurate analysis techniques appropriate to the reduction of this fast ion dynamics data. Moment analysis and curve-fitting routines have been evaluated for noise sensitivity and robustness. Also presented is an analysis method which correctly separates the flux-surface average of the correlated fluctuations in u and B from the fluctuations due to rigid shifts of the plasma column

Transport and fluctuation studies on RFX

International Nuclear Information System (INIS)

Martini, S.; Martines, E.; Carraro, L.

2003-01-01

The paper highlights the progress made in the modelling and understanding of the mechanisms underlying particle and energy transport in the RFP. A transport analysis has been performed on the perturbed transport coefficients for pellets of different characteristics launched both in standard RFP plasmas and in combination with pulsed poloidal current drive, this latter scenario potentially allowing an efficient fuelling and a good energy confinement. The study has confirmed the importance of large-scale MHD modes in determining the transport properties in the RFP core. As far as confinement in the edge is concerned, the analysis has been focussed on the role played by burst events on the electrostatic particle flux and to the 'intermittent events'. The intermittent events cluster during relaxation processes and such behaviour is more evident close to the magnetic reversal surface. This result confirms an interplay between the small scale edge turbulence and the large scale relaxation events driven by core-resonant tearing modes. (author)

Overview of the applications of cement-based immobilization technologies developed at US DOE facilities

International Nuclear Information System (INIS)

Dole, L.R.

1985-01-01

This paper briefly reviews seven cement-based waste form development programs at six of the US Department of Energy (DOE) sites. These sites have developed a variety of processes that range from producing 25-mm-(1-in.-) diam pellets in a glove box to producing 240-m-(800-ft-) diam grout sheets within the bedding planes of a deep shale formation. These successful applications of cement-based waste forms to the many radioactive waste streams from nuclear facilities bear witness to the flexibility and reliability of this class of immobilization materials. The US DOE sites and their programs are: (1) Oak Ridge National Laboratory (ORNL), Hydrofracture Grout; (2) Hanford, Transportable Grout Facility (TGF); (3) Savannah River Plant (SRP), Nitrate Saltcrete; (4) EG and G Idaho, Process Experimental Pilot Plant (PREPP); (5) Mound Laboratory (ML), Waste Pelletization Process; (6) ORNL, FUETAP Concretes, and (7) Rocky Flats Plant (RFP), Inert Carrier Concrete Process (ICCP). The major issues regarding the application of cement-based waste forms to radioactive waste management problems are also presented. These issues are (1) leachability, (2) radiation stability, (3) thermal stability, (4) phase complexity of the matrix, and (5) effects of the waste stream composition. A cursory review of current research in each of these areas is included along with a discussion of future trends in cement-based waste form developments and applications. 35 refs., 12 figs

Cholesterol-dependent energy transfer between fluorescent proteins-insights into protein proximity of APP and BACE1 in different membranes in Niemann-Pick type C disease cells.

Science.gov (United States)

von Einem, Bjoern; Weber, Petra; Wagner, Michael; Malnar, Martina; Kosicek, Marko; Hecimovic, Silva; Arnim, Christine A F von; Schneckenburger, Herbert

2012-11-26

Förster resonance energy transfer (FRET) -based techniques have recently been applied to study the interactions between β-site APP-cleaving enzyme-GFP (BACE1-GFP) and amyloid precursor protein-mRFP (APP-mRFP) in U373 glioblastoma cells. In this context, the role of APP-BACE1 proximity in Alzheimer's disease (AD) pathogenesis has been discussed. FRET was found to depend on intracellular cholesterol levels and associated alterations in membrane stiffness. Here, NPC1 null cells (CHO-NPC1-/-), exhibiting increased cholesterol levels and disturbed cholesterol transport similar to that observed in Niemann-Pick type C disease (NPC), were used to analyze the influence of altered cholesterol levels on APP-BACE1 proximity. Fluorescence lifetime measurements of whole CHO-wild type (WT) and CHO-NPC1-/- cells (EPI-illumination microscopy), as well as their plasma membranes (total internal reflection fluorescence microscopy, TIRFM), were performed. Additionally, generalized polarization (GP) measurements of CHO-WT and CHO-NPC1-/- cells incubated with the fluorescence marker laurdan were performed to determine membrane stiffness of plasma- and intracellular-membranes. CHO-NPC1-/- cells showed higher membrane stiffness at intracellular- but not plasma-membranes, equivalent to cholesterol accumulation in late endosomes/lysosomes. Along with higher membrane stiffness, the FRET efficiency between BACE1-GFP and APP-mRFP was reduced at intracellular membranes, but not within the plasma membrane of CHO-NPC1-/-. Our data show that FRET combined with TIRF is a powerful technique to determine protein proximity and membrane fluidity in cellular models of neurodegenerative diseases.

Cyclin B1 Destruction Box-Mediated Protein Instability: The Enhanced Sensitivity of Fluorescent-Protein-Based Reporter Gene System

Directory of Open Access Journals (Sweden)

Chao-Hsun Yang

2013-01-01

Full Text Available The periodic expression and destruction of several cyclins are the most important steps for the exact regulation of cell cycle. Cyclins are degraded by the ubiquitin-proteasome system during cell cycle. Besides, a short sequence near the N-terminal of cyclin B called the destruction box (D-box; CDB is also required. Fluorescent-protein-based reporter gene system is insensitive to analysis because of the overly stable fluorescent proteins. Therefore, in this study, we use human CDB fused with both enhanced green fluorescent protein (EGFP at C-terminus and red fluorescent protein (RFP, DsRed at N-terminus in the transfected human melanoma cells to examine the effects of CDB on different fluorescent proteins. Our results indicated that CDB-fused fluorescent protein can be used to examine the slight gene regulations in the reporter gene system and have the potential to be the system for screening of functional compounds in the future.

Improved particle confinement in transition from multiple-helicity to quasi-single-helicity regimes of a reversed-field pinch.

Science.gov (United States)

Frassinetti, L; Predebon, I; Koguchi, H; Yagi, Y; Hirano, Y; Sakakita, H; Spizzo, G; White, R B

2006-10-27

The quasi-single-helicity (QSH) state of a reversed-field pinch (RFP) plasma is a regime in which the RFP configuration can be sustained by a dynamo produced mainly by a single tearing mode and in which a helical structure with well-defined magnetic flux surfaces arises. In this Letter, we show that spontaneous transitions to the QSH regime enhance the particle confinement. This improvement is originated by the simultaneous and cooperative action of the increase of the magnetic island and the reduction of the magnetic stochasticity.

Theoretical reversed field pinch studies: Progress report, August 31, 1987 to June 1988

International Nuclear Information System (INIS)

1988-01-01

This paper describes the progress made in Grant DE-FG02-85ER53212 since the end of the last year, August 31, 1987 to the present (June, 1988). Substantial results have emerged in two areas of high importance to the RFP program - nonlinear evolution with nonideal boundaries and self-consistent equilibrium in the presence of field errors. Both of these topics are critical for a basic understanding of RFP physics, for interpretation of current experiments, and for design of future devices and reactors. 3 refs

No need to be HAMLET or BAMLET to interact with histones: binding of monomeric alpha-lactalbumin to histones and basic poly-amino acids.

Science.gov (United States)

Permyakov, Serge E; Pershikova, Irina V; Khokhlova, Tatyana I; Uversky, Vladimir N; Permyakov, Eugene A

2004-05-18

The ability of a specific complex of human alpha-lactalbumin with oleic acid (HAMLET) to induce cell death with selectivity for tumor and undifferentiated cells was shown recently to be mediated by interaction of HAMLET with histone proteins irreversibly disrupting chromatin structure [Duringer, C., et al. (2003) J. Biol. Chem. 278, 42131-42135]. Here we show that monomeric alpha-lactalbumin (alpha-LA) in the absence of fatty acids is also able to bind efficiently to the primary target of HAMLET, histone HIII, regardless of Ca(2+) content. Thus, the modification of alpha-LA by oleic acid is not required for binding to histones. We suggest that interaction of negatively charged alpha-LA with the basic histone stabilizes apo-alpha-LA and destabilizes the Ca(2+)-bound protein due to compensation for excess negative charge of alpha-LA's Ca(2+)-binding loop by positively charged residues of the histone. Spectrofluorimetric curves of titration of alpha-LA by histone H3 were well approximated by a scheme of cooperative binding of four alpha-LA molecules per molecule of histone, with an equilibrium dissociation constant of 1.0 microM. Such a stoichiometry of binding implies that the binding process is not site-specific with respect to histone and likely is driven by just electrostatic interactions. Co-incubation of positively charged poly-amino acids (poly-Lys and poly-Arg) with alpha-LA resulted in effects which were similar to those caused by histone HIII, confirming the electrostatic nature of the alpha-LA-histone interaction. In all cases that were studied, the binding was accompanied by aggregation. The data indicate that alpha-lactalbumin can be used as a basis for the design of antitumor agents, acting through disorganization of chromatin structure due to interaction between alpha-LA and histone proteins.

Rocky Flats Compliance Program

International Nuclear Information System (INIS)

1994-02-01

The Department of Energy (DOE) established the Office of Technology Development (EM-50) (OTD) as an element of Environmental Restoration and Waste Management (EM) in November 1989. The primary objective of the Office of Technology Development, Rocky Flats Compliance Program (RFCP), is to develop altemative treatment technologies for mixed low-level waste (wastes containing both hazardous and radioactive components) to use in bringing the Rocky Flats Plant (RFP) into compliance with Federal and state regulations and agreements. Approximately 48,000 cubic feet of untreated low-level mixed waste, for which treatment has not been specified, are stored at the RFP. The cleanup of the Rocky Flats site is driven by agreements between DOE, the Environmental Protection Agency (EPA), and the Colorado Department of Health (CDH). Under these agreements, a Comprehensive Treatment and Management Plan (CTMP) was drafted to outline the mechanisms by which RFP will achieve compliance with the regulations and agreements. This document describes DOE's strategy to treat low-level mixed waste to meet Land Disposal Restrictions and sets specific milestones related to the regulatory aspects of technology development. These milestones detail schedules for the development of technologies to treat all of the mixed wastes at the RFP. Under the Federal Facilities Compliance Act (FFCA), the CTMP has been incorporated into Rocky Flats Plant Conceptual Site Treatment Plan (CSTP). The CSTP will become the Rocky Flats Plant site Treatment Plan in 1995 and will supersede the CTMP

Ion heating and MHD dynamo fluctuations in the reversed field pinch

International Nuclear Information System (INIS)

Scime, E.E.

1992-05-01

Ion temperature measurements, time resolved to 10 μs, have been made in the Madison Symmetric Torus (MST) reversed field pinch (RFP) with a five channel charge exchange analyzer. The characteristic anomalously high ion temperature of RFP discharges has been observed in the MST. The evolution of the ion and electron temperature, as well as density and charge exchange power loss, were measured for a series of reproducible discharges. The ion heating expected from collisional processes with the electrons is calculated and shown too small to explain the measured ion temperatures. The charge exchange determined ion temperature is also compared to measurements of the thermally broadened CV 227.1 nm line. The ion temperature, T i ∼ 250 eV for I = 360 kA, increases by more than 100% during discrete dynamo bursts in MST discharges. Magnetic field fluctuations in the range 0.5 endash 5 MHz were also measured during the dynamo bursts. Structure in the fluctuation frequency spectrum at the ion cyclotron frequency appears as the bursts terminate, suggesting that the mechanism of ion heating involves the dissipation of dynamo fluctuations at ion cyclotron frequencies. Theoretical models for ion heating are reviewed and discussed in light of the experimental results. Similar electron heating mechanisms may be responsible for the discrepancy between measured and expected loop voltages in the RFP. The electrons, as well as the ions, may be heated by turbulent mechanisms, and a RFP energy budget including such phenomena is described

Magnetic divertor design for the compact reversed-field pinch reactor

International Nuclear Information System (INIS)

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

1984-01-01

A recently completed design of a pumped-limiter-based Compact Reversed-Field Pinch Reactor is used to estimate for the first time the impact of magnetic divertors. A range of divertor options for the low-toroidal-field RFP is examined, and a design selection is made constrained by consideration of field ripple (magnetic island), blanket displacement, recirculating power, cost, heat flux, and access. Design choices based on diversion of minority (toroidal) field lead to a preference for (poloidally) symmetric or bundle divertor geometries

Fabrication and bonding of thiol-ene-based microfluidic devices

DEFF Research Database (Denmark)

Sikanen, Tiina M; Lafleur, Josiane P.; Moilanen, Maria-Elisa

2013-01-01

In this work, the bonding strength of microchips fabricated by thiol-ene free-radical polymerization was characterized in detail by varying the monomeric thiol/allyl composition from the stoichiometric ratio (1:1) up to 100% excess of thiol (2:1) or allyl (1:2) functional groups. Four different...... properties for each application. Here, a capillary electrophoresis separation is performed to demonstrate the attractive properties of stoichiometric thiol-ene microchips....

Prolinimines: N-Amino-l-Pro-methyl Ester (Hydrazine) Schiff Bases from a Fish Gastrointestinal Tract-Derived Fungus, Trichoderma sp. CMB-F563.

Science.gov (United States)

Mohamed, Osama G; Khalil, Zeinab G; Capon, Robert J

2018-01-19

A rice cultivation of a fish gastrointestinal tract-derived fungus, Trichoderma sp. CMB-F563, yielded natural products incorporating a rare hydrazine moiety, embedded within a Schiff base. Structures inclusive of absolute configurations were assigned to prolinimines A-D (1-4) on the basis of detailed spectroscopic and C 3 Marfey's analysis, as well as biosynthetic considerations, biomimetic total synthesis, and chemical transformations. Of note, monomeric 1 proved to be acid labile and, during isolation, underwent quantitative transformation to dimeric 3 and trimeric 4. Prolinimines are only the second reported natural products incorporating an N-amino-Pro residue, the first to include l-Pro, the first to occur as Schiff bases, and the first to be isolated from a microorganism.

A Prospective One-Year Microbiologic Survey of Combined Pneumonia and Respiratory Failure.

Science.gov (United States)

Fisher, Kristen; Trupka, Tracy; Micek, Scott T; Juang, Paul; Kollef, Marin H

2017-10-01

Pneumonia and respiratory failure are common problems in the intensive care unit (ICU) setting, often occurring together. The relative prevalence of pneumonia types (community acquired, hospital acquired, ventilator associated) and causative pathogens is not well described in patients with respiratory failure. This was a prospective observational cohort study conducted in the medical ICU (34 beds) of Barnes-Jewish Hospital, an academic referral center of 1,300 beds from January 2016-December 2016. All patients who were prospectively adjudicated to have respiratory failure and pneumonia (RFP) regardless of pneumonia type were classified into one of four microbiologic categories: pathogen negative, antibiotic-susceptible pathogen (according to ceftriaxone susceptibility), antibiotic-resistant pathogen, and viruses. The primary outcomes assessed were the hospital mortality rate and inappropriate initial antibiotic therapy (IIAT) for non-viral pathogens. Among 364 consecutive patients with RFP, 63 (17.3%) had organisms that were antibiotic susceptible, 104 (28.6%) had antibiotic-resistant organisms, 118 (32.4%) were pathogen negative, and 79 (21.7%) had viral infections. For these categories, IIAT occurred in 3.2%, 21.2%, 0.8%, and 0, respectively (p < 0.001). Vasopressor-requiring shock was present in 61.9%, 72.1%, 68.6%, and 67.1%, respectively (p = 0.585), and the hospital mortality rates were 27.0%, 48.1%, 31.4%, and 36.7%, respectively (p = 0.020). Multivariable logistic regression analysis identified IIAT as an independent predictor of in-hospital death (adjusted odds ratio 5.28; 95% confidence interval 2.72-10.22; p = 0.012). Male gender, increasing Acute Physiology and Chronic Health Evaluation (APACHE) II scores, greater age, and the presence of shock also predicted death. Microbiologic categorization of patients with RFP suggests that antibiotic-resistant pathogens and viruses are associated with the highest mortality rates. Vasopressor

Structure of the red fluorescent protein from a lancelet (Branchiostoma lanceolatum): a novel GYG chromophore covalently bound to a nearby tyrosine

Energy Technology Data Exchange (ETDEWEB)

Pletnev, Vladimir Z., E-mail: vzpletnev@gmail.com; Pletneva, Nadya V.; Lukyanov, Konstantin A.; Souslova, Ekaterina A.; Fradkov, Arkady F.; Chudakov, Dmitry M.; Chepurnykh, Tatyana; Yampolsky, Ilia V. [Russian Academy of Sciences, Moscow (Russian Federation); Wlodawer, Alexander [National Cancer Institute, Frederick, MD 21702 (United States); Dauter, Zbigniew [National Cancer Institute, Argonne, IL 60439 (United States); Pletnev, Sergei, E-mail: vzpletnev@gmail.com [National Cancer Institute, Argonne, IL 60439 (United States); SAIC-Frederick, Argonne, IL 60439 (United States); Russian Academy of Sciences, Moscow (Russian Federation)

2013-09-01

��70 nm) of laRFP was verified by extensive structure-based site-directed mutagenesis.

Experimental studies of tearing mode and resistive wall mode dynamics in the reversed field pinch configuration

International Nuclear Information System (INIS)

Malmberg, Jenny-Ann

2003-06-01

It is relatively straightforward to establish equilibrium in magnetically confined plasmas, but the plasma is frequently susceptible to a variety of instabilities that are driven by the free energy in the magnetic field or in the pressure gradient. These unstable modes exhibit effects that affect the particle, momentum and heat confinement properties of the configuration. Studies of the dynamics of several of the most important modes are the subject of this thesis. The studies are carried out on plasmas in the reversed field pinch (RFP) configuration. One phenomenon commonly observed in RFPs is mode wall locking. The localized nature of these phase- and wall locked structures results in localized power loads on the wall which are detrimental for confinement. A detailed study of the wall locked mode phenomenon is performed based on magnetic measurements from three RFP devices. The two possible mechanisms for wall locking are investigated. Locking as a result of tearing modes interacting with a static field error and locking due to the presence of a non-ideal boundary. The characteristics of the wall locked mode are qualitatively similar in a device with a conducting shell system (TPE-RX) compared to a device with a resistive shell (Extrap T2). A theoretical model is used for evaluating the threshold values for wall locking due to eddy currents in the vacuum vessel in these devices. A good correlation with experiment is observed for the conducting shell device. The possibility of successfully sustaining discharges in a resistive shell RFP is introduced in the recently rebuilt device Extrap T2R. Fast spontaneous mode rotation is observed, resulting in low magnetic fluctuations, low loop voltage and improved confinement. Wall locking is rarely observed. The low tearing mode amplitudes allow for the theoretically predicted internal non-resonant on-axis resistive wall modes to be observed. These modes have not previously been distinguished due to the formation of wall

Managing the training process: contracting for training services

International Nuclear Information System (INIS)

Carlson, E.R.

1985-01-01

The need for using consultants and contracting for training services should be based on one or more of these three major reasons: the need to expand capability on a crash basis, the need for specialized expertise, and the need for objectivity, and independence. When the need for using a consultant exists, the training manager must first define the scope of work or services the consultant is to provide and how the consultant will interface with the training staff. The next step will be to develop a Request for Proposal which involves management, training staff, and contract services organizations. The critical parts of an RFP are scope and schedule. Consideration would be given to determining which consultant organizations should receive the RFP, and specifying who is on the bidders list. The third step will consist of evaluating the proposals and awarding the work. This step needs to be objective, defendable, and repeatable. The final phase will be the performance of the work. The activities included are project initiation, monitoring contractor performance, and formal conclusion

Mesoscale meteorological measurements characterizing complex flows

International Nuclear Information System (INIS)

Hubbe, J.M.; Allwine, K.J.

1993-09-01

Meteorological measurements are an integral and essential component of any emergency response system for addressing accidental releases from nuclear facilities. An important element of the US Department of Energy's (DOE's) Atmospheric Studies in Complex Terrain (ASCOT) program is the refinement and use of state-of-the-art meteorological instrumentation. ASCOT is currently making use of ground-based remote wind sensing instruments such as doppler acoustic sounders (sodars). These instruments are capable of continuously and reliably measuring winds up to several hundred meters above the ground, unattended. Two sodars are currently measuring the winds, as part of ASCOT's Front Range Study, in the vicinity of DOE's Rocky Flats Plant (RFP) near Boulder, Colorado. A brief description of ASCOT's ongoing Front Range Study is given followed by a case study analysis that demonstrates the utility of the meteorological measurement equipment and the complexity of flow phenomena that are experienced near RFP. These complex flow phenomena can significantly influence the transport of the released material and consequently need to be identified for accurate assessments of the consequences of a release

RNA-ID, a Powerful Tool for Identifying and Characterizing Regulatory Sequences.

Science.gov (United States)

Brule, C E; Dean, K M; Grayhack, E J

2016-01-01

The identification and analysis of sequences that regulate gene expression is critical because regulated gene expression underlies biology. RNA-ID is an efficient and sensitive method to discover and investigate regulatory sequences in the yeast Saccharomyces cerevisiae, using fluorescence-based assays to detect green fluorescent protein (GFP) relative to a red fluorescent protein (RFP) control in individual cells. Putative regulatory sequences can be inserted either in-frame or upstream of a superfolder GFP fusion protein whose expression, like that of RFP, is driven by the bidirectional GAL1,10 promoter. In this chapter, we describe the methodology to identify and study cis-regulatory sequences in the RNA-ID system, explaining features and variations of the RNA-ID reporter, as well as some applications of this system. We describe in detail the methods to analyze a single regulatory sequence, from construction of a single GFP variant to assay of variants by flow cytometry, as well as modifications required to screen libraries of different strains simultaneously. We also describe subsequent analyses of regulatory sequences. © 2016 Elsevier Inc. All rights reserved.

Structure-Based Analysis of Toxoplasma gondii Profilin: A Parasite-Specific Motif Is Required for Recognition by Toll-Like Receptor 11

Energy Technology Data Exchange (ETDEWEB)

K Kucera; A Koblansky; L Saunders; K Frederick; E De La Cruz; S Ghosh; Y Modis

2011-12-31

Profilins promote actin polymerization by exchanging ADP for ATP on monomeric actin and delivering ATP-actin to growing filament barbed ends. Apicomplexan protozoa such as Toxoplasma gondii invade host cells using an actin-dependent gliding motility. Toll-like receptor (TLR) 11 generates an innate immune response upon sensing T. gondii profilin (TgPRF). The crystal structure of TgPRF reveals a parasite-specific surface motif consisting of an acidic loop, followed by a long {beta}-hairpin. A series of structure-based profilin mutants show that TLR11 recognition of the acidic loop is responsible for most of the interleukin (IL)-12 secretion response to TgPRF in peritoneal macrophages. Deletion of both the acidic loop and the {beta}-hairpin completely abrogates IL-12 secretion. Insertion of the T. gondii acidic loop and {beta}-hairpin into yeast profilin is sufficient to generate TLR11-dependent signaling. Substitution of the acidic loop in TgPRF with the homologous loop from the apicomplexan parasite Cryptosporidium parvum does not affect TLR11-dependent IL-12 secretion, while substitution with the acidic loop from Plasmodium falciparum results in reduced but significant IL-12 secretion. We conclude that the parasite-specific motif in TgPRF is the key molecular pattern recognized by TLR11. Unlike other profilins, TgPRF slows nucleotide exchange on monomeric rabbit actin and binds rabbit actin weakly. The putative TgPRF actin-binding surface includes the {beta}-hairpin and diverges widely from the actin-binding surfaces of vertebrate profilins.

RFP # 13140013: Request for proposals: Improving development ...

International Development Research Centre (IDRC) Digital Library (Canada)

Teisha Youden

public-private partnerships. Questions and Answers # 2. 1. Is there any specification as to how many countries and which countries shall be covered under the study ... any PPP that benefits SMEs, or otherwise? RESPONSE: Here, there is room for the proponent to tell us how SMEs are affected by or can participate in PPPs.

Photomonomerization of pyrimidine dimers by indoles and proteins

Energy Technology Data Exchange (ETDEWEB)

Chen, J.; Huang, C.W.; Hinman, L.; Gordon, M.P.; Deranleau, D.A.

1976-01-01

Model systems for the study of photoreactivation have been developed that utilize a variety of indole derivatives. These systems can split uracil cis-syn cyclobutadipyrimidine, either free or in RNA, when irradiated at wavelengths absorbed only by the indole moiety. The ability of indole compounds to split dimers is closely related to their electronic properties. Those of high electron-donor capacity such as indole, 3-methylindole, indole-3-acetic acid, 5-hydroxytryptophan and tryptophan are good photosensitizers, with efficacy in that order. Indoles with electron-withdrawing substituents such as indole-3-carboxylic acid, indole-3-aldehyde and oxindole are inactive in the monomerization reaction. These findings support the proposed mechanism that the photosensitized monomerization occurs as a result of electron transfer from the excited indole molecules to the pyrimidine bases. Proteins containing fully exposed tryptophan residues (chicken egg white lysozyme and bovine diisopropylphosphoryltrypsin) also cause the splitting of the /sup 14/C-labeled dimers under the same conditions. In the case of lysozyme the quantum yield of monomerization is similar to that of free tryptophan. Much of the monomerization ability of lysozyme was lost after the solvent-available tryptophan had been oxidized by treatment with N-bromosuccinimide. Bovine pancreatic ribonuclease A, a protein devoid of tryptophan, failed to exhibit photosensitized monomerization of uracil dimers. The biological implication of these reactions involving a protein with an exposed tryptophan residue is discussed. Although indoles are able to split the dimers in RNA, they fail to photoreactivate uv-damaged TMV-RNA. Indole-3-acetic acid, 3-methylindole and 5-hydroxytryptophan rapidly inactive viral RNA when irradiated at 313 nm, possibly because of side reactions.

A Knockin Reporter Allows Purification and Characterization of mDA Neurons from Heterogeneous Populations

Directory of Open Access Journals (Sweden)

Ninuo Xia

2017-03-01

Full Text Available Generation of midbrain dopaminergic (mDA neurons from human pluripotent stem cells provides a platform for inquiry into basic and translational studies of Parkinson’s disease (PD. However, heterogeneity in differentiation in vitro makes it difficult to identify mDA neurons in culture or in vivo following transplantation. Here, we report the generation of a human embryonic stem cell (hESC line with a tyrosine hydroxylase (TH-RFP (red fluorescent protein reporter. We validated that RFP faithfully mimicked TH expression during differentiation. Use of this TH-RFP reporter cell line enabled purification of mDA-like neurons from heterogeneous cultures with subsequent characterization of neuron transcriptional and epigenetic programs (global binding profiles of H3K27ac, H3K4me1, and 5-hydroxymethylcytosine [5hmC] at four different stages of development. We anticipate that the tools and data described here will contribute to the development of mDA neurons for applications in disease modeling and/or drug screening and cell replacement therapies for PD.

Tokamak-like confinement at high beta and low field in the reversed field pinch

International Nuclear Information System (INIS)

Sarff, J S; Anderson, J K; Biewer, T M; Brower, D L; Chapman, B E; Chattopadhyay, P K; Craig, D; Deng, B; Hartog, D J Den; Ding, W X; Fiksel, G; Forest, C B; Goetz, J A; O'Connell, R; Prager, S C; Thomas, M A

2003-01-01

For several reasons, improved-confinement achieved in the reversed field pinch (RFP) during the last few years can be characterized as 'tokamak-like'. Historically, RFP plasmas have had relatively poor confinement due to tearing instability which causes magnetic stochasticity and enhanced transport. Tearing reduction is achieved through modification of the inductive current drive, which dramatically improves confinement. The electron temperature increases to >1 keV and the electron heat diffusivity decreases to approx. 5 m 2 s -1 , comparable with the transport level expected in a tokamak plasma of the same size and current. This corresponds to a 10-fold increase in global energy confinement. Runaway electrons are confined, and Fokker-Planck modelling of the electron distribution reveals that the diffusion at high energy is independent of the parallel velocity, uncharacteristic of stochastic transport. Improved-confinement occurs simultaneously with increased beta approx. 15%, while maintaining a magnetic field strength ten times weaker than a comparable tokamak. Measurements of the current, magnetic, and electric field profiles show that a simple Ohm's Law applies to this RFP sustained without dynamo relaxation

A Knockin Reporter Allows Purification and Characterization of mDA Neurons from Heterogeneous Populations.

Science.gov (United States)

Xia, Ninuo; Fang, Fang; Zhang, Pengbo; Cui, Jun; Tep-Cullison, Chhavy; Hamerley, Tim; Lee, Hyun Joo; Palmer, Theo; Bothner, Brian; Lee, Jin Hyung; Pera, Renee Reijo

2017-03-07

Generation of midbrain dopaminergic (mDA) neurons from human pluripotent stem cells provides a platform for inquiry into basic and translational studies of Parkinson's disease (PD). However, heterogeneity in differentiation in vitro makes it difficult to identify mDA neurons in culture or in vivo following transplantation. Here, we report the generation of a human embryonic stem cell (hESC) line with a tyrosine hydroxylase (TH)-RFP (red fluorescent protein) reporter. We validated that RFP faithfully mimicked TH expression during differentiation. Use of this TH-RFP reporter cell line enabled purification of mDA-like neurons from heterogeneous cultures with subsequent characterization of neuron transcriptional and epigenetic programs (global binding profiles of H3K27ac, H3K4me1, and 5-hydroxymethylcytosine [5hmC]) at four different stages of development. We anticipate that the tools and data described here will contribute to the development of mDA neurons for applications in disease modeling and/or drug screening and cell replacement therapies for PD. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

Two-fluid and nonlinear effects of tearing and pressure-driven resistive modes in reversed field pinches

International Nuclear Information System (INIS)

Mirnov, V.V.

2002-01-01

Large-scale tearing instabilities have long been considered to underlie transport and dynamo processes in the reversed field pinch (RFP). The vast majority of theoretical and computational RFP work has focused on pressureless, single-fluid MHD in cylindrical plasmas driven solely by a toroidal electric field. We report results of five investigations covering two-fluid dynamos, toroidal nonlinear MHD computation, nonlinear computation of Oscillating Field Current Drive (OFCD), the effect of shear flow on tearing instability, and the effect of pressure on resistive instability. The key findings are: (1) two-fluid dynamo arising from the Hall term is much larger than the standard MHD dynamo present in a single-fluid treatment, (2) geometric coupling from toroidicity precludes the occurrence of laminar single helicity states, except for nonreversed plasmas, (3) OFCD, a form of AC helicity injection, can sustain the RFP plasma current, although magnetic fluctuations are enhanced, (4) edge shear flow can destabilize the edge resonant m=0 modes, which occur as spikes in experiment, and (5) pressure driven modes are resistive at low beta, only becoming ideal at extremely high beta. (author)

Equilibrium poloidal field distributions in reversed-field-pinch toroidal discharges

International Nuclear Information System (INIS)

Baker, D.A.; Mann, L.W.; Schoenberg, K.F.

1982-04-01

A comparison between the analytic formulae of Shafranov for equilibrium in axisymmetric toroidal reversed field pinch (RFP) systems and fully toroidal numerical solutions of the Grad-Shafranov equation is presented as a function of poloidal beta, internal plasma inductance, and aspect ratio. The Shafranov formula for the equilibrium poloidal field distribution is accurate to within 5% for aspect ratios greater than 2, poloidal betas less than 50%, and for plasma current channels that exceed one-third of the minor toroidal radius. The analytic description for the center shift of the innermost flux surface that encloses the plasma current (the Shafranov shift) is accurate to within 15% for aspect ratios greater than 2 and poloidal betas below 50%, provided the shift does not exceed one-tenth of the minor conducting boundary radius. The behavior of the magnetic axis shift as a function of plasma parameters is included. The Shafranov formulae provide a convenient method for describing the equilibrium behavior of an RFP discharge. Examples illustrating the application of the analytic formulae to the Los Alamos ZT-40M RFP experiment are given

Equilibrium poloidal-field distributions in reversed-field-pinch toroidal discharges

International Nuclear Information System (INIS)

Baker, D.A.; Mann, L.W.; Schoenberg, K.F.

1983-01-01

A comparison between the approximate analytic formulae of Shafranov for equilibrium in axisymmetric toroidal systems and fully toroidal numerical solutions of the Grad-Shafranov equation for reversed-field-pinch (RFP) configurations is presented as a function of poloidal beta, internal plasma inductance, and aspect ratio. The Shafranov formula for the equilibrium poloidal-field distribution at the conducting shell that surrounds the plasma is accurate to within 5% for aspect ratios greater than 2, poloidal betas less than 50%, and for plasma current channels that exceed one third of the minor toroidal radius. The analytic description for the centre shift of the innermost flux surface that encloses the plasma current (the Shafranov shift) is accurate to within 15% for aspect ratios greater than 2 and poloidal betas below 50%, provided the shift does not exceed one tenth of the minor conducting boundary radius. The Shafranov formulae provide a convenient method for describing the gross equilibrium behaviour of an axisymmetric RFP discharge, as well as an effective tool for designing the poloidal-field systems of RFP experiments. (author)

The Rocky Flats Plant Waste Stream and Residue Identification and Characterization Program (WSRIC): Progress and achievements

International Nuclear Information System (INIS)

Ideker, V.L.

1994-01-01

The Waste Stream and Residue Identification and Characterization (WSRIC) Program, as described in the WSRIC Program Description delineates the process knowledge used to identify and characterize currently-generated waste from approximately 5404 waste streams originating from 576 processes in 288 buildings at Rocky Flats Plant (RFP). Annual updates to the WSRIC documents are required by the Federal Facilities Compliance Agreement between the US Department of Energy, the Colorado Department of Health and the Environmental Protection Agency. Accurate determination and characterization of waste is a crucial component in RFP's waste management strategy to assure compliance with Resource Conservation and Recovery Act (RCRA) storage and treatment requirements, as well as disposal acceptance criteria. The WSRIC Program was rebaselined in September 1992, and serves as the linchpin for documenting process knowledge in RFP's RCRA operating record. Enhancements to the WSRIC include strengthening the waste characterization rationale, expanding WSRIC training for waste generators, and incorporating analytical information into the WSRIC building books. These enhancements will improve credibility with the regulators and increase waste generators' understanding of the basis for credible waste characterizations

Quantitative plasma spectroscopy at JET and Extrap-T1

International Nuclear Information System (INIS)

Zastrow, K.D.

1993-01-01

Studies in quantitative plasma spectroscopy are performed on the Joint European Torus (JET) in Culham, Great-Britain and on the Extrap-T1 reversed-field pinch (RFP) in Stockholm. The model concepts that form the basis of these studies are reviewed. At JET, spectra of He-like nickel are observed with a high-resolution X-ray crystal spectrometer. The experimental line intensity ratios of satellite lines to the resonance line are compared with theoretical data. The agreement is found to be good, with the exception of the excitation of dipole-forbidden lines. The spectrum is also used to derive central ion temperature, central toroidal rotation and nickel concentration based upon a model for the radial emission. The results are compared with those from an independent diagnostic, charge-exchange recombination spectroscopy (CWRS). Theoretically predicted cross section effects on the CXRS data are verified. On Extrap-T1, vacuum ultraviolet (VUV) spectra and visible spectra are analysed. From these, thermodynamic quantities of the plasma are derived, like electron temperature, impurity concentrations and particle fluxes. The oxygen ionization balance is measured and compared to calculations with a collisional-dielectronic model with metastable resolution, both in 0-dimensional time-dependent and transport model calculations. The performance of the RFP discharges investigated in terms of radiative power loss and energy and particle confinement properties. The scaling of the energy confinement time with plasma current, pinch parameter and electron density is found to be dominated by the dynamo activity needed to sustain the RFP configuration. The scaling of the particle confinement time, on the other hand, is dominated by pressure-driven activity associated with the regulation of β

Monomeric, dimeric and multimeric system of RGD peptides radiolabeled with 177Lu for tumors therapy that expressing αβ integrin s

International Nuclear Information System (INIS)

Luna G, M. A.

2014-01-01

The conjugation of peptides to gold nanoparticles (AuNPs) produces biocompatible and stable multimeric systems with target-specific molecular recognition. Peptides based on the cyclic Arg-Gly-Asp (RGD) sequence have been reported as high affinity agents for the α(v)β(3) and α(v)β(5) integrin. The aim of this research was to prepare a multimeric system of 177 Lu-labeled gold nanoparticles conjugated to c[RGDfK(C)] [cyclo(Arg-Gly-Asp-Phe-Lys(Cys)] peptides and to compare the radiation absorbed dose with that of 177 Lu-labeled monomeric and dimeric RGD peptides to α(v)β(3) integrin-positive U87MG tumors in mice, as well as, evaluate the in vitro potential 177 Lu-AuNP-c[RGDfK(C)] as a plasmonic photothermal therapy and targeted radiotherapy system in MCF7 breast cancer cells. DOTA-GGC (1,4,7,10-tetraaza cyclododecane-N,N,N-tetraacetic-Gly-Gly-Cys) and c[RGDfK(C)] peptides were synthesized and conjugated to AuNPs by the spontaneous reaction of the thiol groups. Tem, UV-Vis, XP S, Raman and Far-IR spectroscopy techniques demonstrated that AuNPs were functionalized with the peptides. To obtain 177 Lu-AuNP-c[RGDfK(C)], the 177 Lu-DOTA-GGC radio peptide was first prepared and added to a solution of AuNPs followed by c[RGDfK(C)] (25 μL, 5 μM) at 18 grades C for 15 min. 177 Lu-DOTA-GGC, 177 Lu- DOTA-cRGDfK and 177 Lu-DOTA-E-c(RGDfK) 2 were prepared by adding 177 LuCl 3 (370 MBq) to 5 μL (1 mg/ml) of the DOTA derivative diluted with 50 μL of 1 M acetate buffer at ph 5. The mixture was incubated at 90 grades C in a block heater for 30 min. Radiochemical purity was determined by ultrafiltration and HPLC analyses. After laser irradiation, the presence of c[RGDfK(C)]-AuNP in cells caused a significant increase in the temperature of the medium (50.5 grades C, compared to 40.3 grades C without AuNPs) resulting in a significant decrease in MCF7 cell viability down to 9 %. After treatment with 177 Lu-AuNP-c[RGDfK(C)], the MCF7 cell proliferation was significantly inhibited

Specialized mouse embryonic stem cells for studying vascular development.

Science.gov (United States)

Glaser, Drew E; Burns, Andrew B; Hatano, Rachel; Medrzycki, Magdalena; Fan, Yuhong; McCloskey, Kara E

2014-01-01

Vascular progenitor cells are desirable in a variety of therapeutic strategies; however, the lineage commitment of endothelial and smooth muscle cell from a common progenitor is not well-understood. Here, we report the generation of the first dual reporter mouse embryonic stem cell (mESC) lines designed to facilitate the study of vascular endothelial and smooth muscle development in vitro. These mESC lines express green fluorescent protein (GFP) under the endothelial promoter, Tie-2, and Discomsoma sp. red fluorescent protein (RFP) under the promoter for alpha-smooth muscle actin (α-SMA). The lines were then characterized for morphology, marker expression, and pluripotency. The mESC colonies were found to exhibit dome-shaped morphology, alkaline phosphotase activity, as well as expression of Oct 3/4 and stage-specific embryonic antigen-1. The mESC colonies were also found to display normal karyotypes and are able to generate cells from all three germ layers, verifying pluripotency. Tissue staining confirmed the coexpression of VE (vascular endothelial)-cadherin with the Tie-2 GFP+ expression on endothelial structures and smooth muscle myosin heavy chain with the α-SMA RFP+ smooth muscle cells. Lastly, it was verified that the developing mESC do express Tie-2 GFP+ and α-SMA RFP+ cells during differentiation and that the GFP+ cells colocalize with the vascular-like structures surrounded by α-SMA-RFP cells. These dual reporter vascular-specific mESC permit visualization and cell tracking of individual endothelial and smooth muscle cells over time and in multiple dimensions, a powerful new tool for studying vascular development in real time.

Active control of multiple resistive wall modes

International Nuclear Information System (INIS)

Brunsell, P. R.; Yadikin, D.; Gregoratto, D.; Paccagnella, R.; Liu, Y. Q.; Bolzonella, T.; Cecconello, M.; Drake, J. R.; Kuldkepp, M.; Manduchi, G.; Marchiori, G.; Marrelli, L.; Partin, P.; Menmuir, S.; Ortolani, S.; Rachlew, E.; Spizzo, S.; Zanca, P.

2005-01-01

Active magnetic feedback suppression of resistive wall modes is of common interest for several fusion concepts relying on close conducting walls for stabilization of ideal magnetohydrodynamic (MHD) modes. In the advanced tokamak without plasma rotation the kink mode is not completely stabilized, but rather converted into an unstable resistive wall mode (RWM) with a growth time comparable to the wall magnetic flux penetration time. The reversed field pinch (RFP) is similar to the advanced tokamak in the sense that it uses a conducting wall for kink mode stabilization. Also both configurations are susceptible to resonant field error amplification of marginally stable modes. However, the RFP has a different RWM spectrum and, in general, a range of modes is unstable. Hence, the requirement for simultaneous feedback stabilization of multiple independent RWMs arises for the RFP configuration. Recent experiments on RWM feedback stabilization, performed in the RFP device EXTRAP T2R [1], are presented. The experimental results obtained are the first demonstration of simultaneous feedback control of multiple independent RWMs [2]. Using an array of active magnetic coils, a reproducible suppression of several RWMs is achieved for the duration of the discharge, 3-5 wall times, through feedback action. An array with 64 active saddle coils at 4 poloidal times 16 toroidal positions is used. The important issues of side band generation by the active coil array and the accompanying coupling of different unstable modes through the feedback action are addressed in this study. Open loop control experiments have been carried out to quantitatively study resonant field error amplification. (Author)

3-dimensional simulation of dynamo effect of reversed field pinch

International Nuclear Information System (INIS)

Koide, Shinji.

1990-09-01

A non-linear numerical simulation of the dynamo effect of a reversed field pinch (RFP) with finite beta is presented. It is shown that the m=-1, n=(9,10,11,....,19) modes cause the dynamo effect and sustain the field reversed configuration. The role of the m=0 modes on the dynamo effect is carefully examined. Our simulation shows that the magnetic field fluctuation level scales as S -0.2 or S -0.3 in the range of 10 3 5 , while Nebel, Caramana and Schnack obtained the fluctuation level is independent of S for a pressureless RFP plasma. (author)

Reversed-field pinch configuration with minimum energy and finite beta

International Nuclear Information System (INIS)

Zhang Peng

1989-01-01

The reversed-field pinch (RFP) configuration has been studied for the case of finite beta. Suydam's condition and the sufficient criterion have been used to examine this configuration. Results of numerical calculations show that the critical value of the pinch parameter Θ for the appearance of the reverse toroidal field increases as the β-value increases. The critical value of Θ for the helical state increases with β as well. Suydam's and Robinson's stability regions increase and shift towards higher values of Θ with increasing β. Theoretical results for finite β coincide with recent RFP experimental results

Ion heat conduction losses in Extrap

International Nuclear Information System (INIS)

Tennfors, E.

1989-08-01

The classical ion heat conduction losses in Extrap discharges are calculated using polynomial magnetic field profiles and compared to the power input. For polynomials matched to magnetic field profiles measured in present experiments, these losses are small. By varying the coefficients of the polynomials, a region is found, where the power input can balance the classical heat conduction losses. Each set of coefficients corresponds to values of the parameters F and Θ used in RFP physics. The region determines a region in an F-Θ diagram, including the usual RFP region but extending to higher values of Θ and βΘ

Market driven strategy for acquisition of waste acceptance and transportation services for commercial spent fuel in the United States

International Nuclear Information System (INIS)

Lemeshewky, W.; Macaluso, C.; Smith, P.; Teer, B.

1998-05-01

The Department of Energy has the responsibility for the shipment of spent nuclear fuel (SNF) from commercial reactors to a Federal facility for storage and/or disposal. DOE has developed a strategy for a market driven approach for the acquisition of transportation services and equipment which will maximize the participation of private industry. To implement this strategy, DOE is planning to issue a Request for Proposal (RFP) for the provision of the required services and equipment to accept SNF from the utilities and transport the SNF to a Federal facility. The paper discusses this strategy and describes the RFP

Numerical studies of active current profile control in the reversed-field pinch

International Nuclear Information System (INIS)

Dahlin, J-E; Scheffel, J; Anderson, J K

2007-01-01

Quenching of the reversed-field pinch (RFP) dynamo is observed in numerical simulations using current profile control. A novel algorithm employing active feedback of the dynamo field has been utilized. The quasi-steady state achieved represents an important improvement as compared with earlier numerical work and may indicate a direction for the design of future experiments. Both earlier and the novel schemes of feedback control result in quasi-single helicity states. The energy confinement time and poloidal beta are observed to be substantially increased, as compared with the conventional RFP, in both the cases. Different techniques for experimental implementation are discussed

Quantification of equine immunoglobulin A in serum and secretions by a fluorescent bead-based assay.

Science.gov (United States)

Schnabel, Christiane L; Babasyan, Susanna; Freer, Heather; Wagner, Bettina

2017-06-01

Only few quantitative reports exist about the concentrations and induction of immunoglobulin A (IgA) in mucosal secretions of horses. Despite this, it is widely assumed that IgA is the predominant immunoglobulin on mucosal surfaces in the horse. Here, two new monoclonal antibodies (mAbs) against equine IgA, clones 84-1 and 161-1, were developed and characterized in detail. Both IgA mAbs specifically bound monomeric and dimeric equine IgA in different applications, such as Western blots and fluorescent bead-based assays. Cross-reactivity with other equine immunoglobulin isotypes was not observed. The new IgA mAb 84-1 was used in combination with the previously characterized anti-equine IgA mAb BVS2 for the development and validation of a fluorescent bead-based assay to quantify total IgA in equine serum and various secretions. The IgA assay's linear detection ranged from 64pg/ml to 1000ng/ml. For the quantification of IgA in serum or in secretions an IgA standard was purified from serum or nasal wash fluid (secretory IgA), respectively. The different standards were needed for accurate IgA quantification in the respective samples taking the different signal intensities of monomeric and dimeric IgA on the florescent bead-based assay into account. IgA was quantified by the bead-based assay established here in different equine samples of healthy adult individuals. In serum the median total IgA was 0.45mg/ml for Thoroughbred horses (TB, n=10) and 1.16mg/ml in Icelandic horses (ICH, n=12). In nasopharyngeal secretions of TB (n=7) 0.13mg/ml median total IgA was measured, and 0.25mg/ml for ICH (n=12). Saliva of ICH (n=6) contained a median of 0.15mg/ml, colostrum of Warmbloods (n=8) a median of 1.89mg/ml IgA. Compared to IgG1 and IgG4/7 quantified in the same samples, IgA appeared as the major immunoglobulin isotype in nasopharyngeal secretions and saliva while it is a minor isotype in serum and colostrum. The newly developed monoclonal antibodies against equine IgA and the

Carbamylated monomeric allergoids as a therapeutic option for sublingual immunotherapy of dust mite- and grass pollen-induced allergic rhinoconjunctivitis: a systematic review of published trials with a meta-analysis of treatment using Lais® tablets.

Science.gov (United States)

Mösges, R; Ritter, B; Kayoko, G; Allekotte, S

2010-10-01

Lais® allergoid tablets contain allergens that are modified by carbamylation. Due to their modified chemical structure, they are suitable for sublingual immunotherapy (SLIT) (13, 16, 17, 24). Based on their small molecule size of 12 to 40 kDa, they can be easily absorbed via the oral mucosa (1). In this review, we studied the efficacy of SLIT with carbamylated monomeric allergoid tablets in the treatment of grass pollen- and dust mite-induced allergic rhinoconjunctivitis on the basis of symptom and medication score improvements. Following a selective internet and databank search, six trials-some placebo-controlled-regarding the treatment of grass pollen- (n = 266) and dust mite-induced (n = 241) allergic rhinoconjunctivitis were used to draw conclusions regarding the clinical efficacy of allergoid tablets. The primary endpoints in these trials were decreases in the need for allergy medications and/or reductions in the occurrence of rhinoconjunctivitis symptoms. Data was recorded from patient diaries regarding their symptoms and medications used and conclusions were then drawn about the effectiveness and tolerabieity of Lais® tablets. The average improvement in symptom score in three trials of grass pollen allergy treatment was 34% in comparison to the placebo group. The treatment of dust mite-induced rhinoconjunctivitis produced an average symptom score improvement of 22% compared to the placebo or control groups. The intake of symptomatic rescue medication during allergoid tablet therapy declined. Treatment of grass pollen allergies and dust mite-induced rhinoconjunctivitis showed an average medication score improvement of 49% and 24%, respectively. Few side effects were documented in the trials and predominantly local effects were observed. Severe systemic side effects did not occur. On the basis of the trial results summarized in this review, we suggest that SLIT using Lais® sublingual tablets is an effective and well-tolerated form of treatment.

Deconstruction of ionic liquid pretreated lignocellulosic biomass using mono-component cellulases and hemicellulases and commercial mixtures

Science.gov (United States)

Lignocellulosic biomass is comprised of cellulose and hemicellulose, sources of polysaccharides, and lignin, a macromolecule with extensive aromaticity. Lignocellulose requires pretreatment before biochemical conversion to its monomeric sugars which can provide a renewable carbon based feedstock for...

Intracellular cellobiose metabolism and its applications in lignocellulose-based biorefineries.

Science.gov (United States)

Parisutham, Vinuselvi; Chandran, Sathesh-Prabu; Mukhopadhyay, Aindrila; Lee, Sung Kuk; Keasling, Jay D

2017-09-01

Complete hydrolysis of cellulose has been a key characteristic of biomass technology because of the limitation of industrial production hosts to use cellodextrin, the partial hydrolysis product of cellulose. Cellobiose, a β-1,4-linked glucose dimer, is a major cellodextrin of the enzymatic hydrolysis (via endoglucanase and exoglucanase) of cellulose. Conversion of cellobiose to glucose is executed by β-glucosidase. The complete extracellular hydrolysis of celluloses has several critical barriers in biomass technology. An alternative bioengineering strategy to make the bioprocessing less challenging is to engineer microbes with the abilities to hydrolyze and assimilate the cellulosic-hydrolysate cellodextrin. Microorganisms engineered to metabolize cellobiose rather than the monomeric glucose can provide several advantages for lignocellulose-based biorefineries. This review describes the recent advances and challenges in engineering efficient intracellular cellobiose metabolism in industrial hosts. This review also describes the limitations of and future prospectives in engineering intracellular cellobiose metabolism. Copyright © 2017 Elsevier Ltd. All rights reserved.

Magnetic fluctuation driven cross-field particle transport in the reversed-field pinch

International Nuclear Information System (INIS)

Scheffel, J.; Liu, D.

1997-01-01

Electrostatic and electromagnetic fluctuations generally cause cross-field particle transport in confined plasmas. Thus core localized turbulence must be kept at low levels for sufficient energy confinement in magnetic fusion plasmas. Reversed-field pinch (RFP) equilibria can, theoretically, be completely stable to ideal and resistive (tearing) magnetohydrodynamic (MHD) modes at zero beta. Unstable resistive interchange modes are, however, always present at experimentally relevant values of the poloidal beta β θ . An analytical quasilinear, ambipolar diffusion model is here used to model associated particle transport. The results indicate that core density fluctuations should not exceed a level of about 1% for plasmas of fusion interest. Parameters of experimentally relevant stationary states of the RFP were adjusted to minimize growth rates, using a fully resistive linearized MHD stability code. Density gradient effects are included through employing a parabolic density profile. The scaling of particle diffusion [D(r)∝λ 2 n 0.5 T/aB, where λ is the mode width] is such that the effects of particle transport are milder in present day RFP experiments than in future reactor-relevant plasmas. copyright 1997 American Institute of Physics

Point design for deuterium-deuterium compact reversed-field pinch reactors

International Nuclear Information System (INIS)

Dabiri, A.E.; Dobrott, D.R.; Gurol, H.; Schnack, D.D.

1984-01-01

A deuterium-deuterium (D-D) reversed-field pinch (RFP) reactor may be made comparable in size and cost to a deuterium-tritium (D-T) reactor at the expense of high-thermal heat load to the first wall. This heat load is the result of the larger percentage of fusion power in charged particles in the D-D reaction as compared to the D-T reaction. The heat load may be reduced by increasing the reactor size and hence the cost. In addition to this ''degraded'' design, the size may be kept small by means of a higher heat load wall, or by means of a toroidal divertor, in which case most of the heat load seen by the wall is in the form of radiation. Point designs are developed for these approaches and cost studies are performed and compared with a D-T reactor. The results indicate that the cost of electricity of a D-D RFP reactor is about20% higher than a D-T RFP reactor. This increased cost could be offset by the inherent safety features of the D-D fuel cycle

Chaos in plasma simulation and experiment

International Nuclear Information System (INIS)

Watts, C.; Sprott, J.C.

1993-09-01

We investigate the possibility that chaos and simple determinism are governing the dynamics of reversed field pinch (RFP) plasmas using data from both numerical simulations and experiment. A large repertoire of nonlinear analysis techniques is used to identify low dimensional chaos. These tools include phase portraits and Poincard sections, correlation dimension, the spectrum of Lyapunov exponents and short term predictability. In addition, nonlinear noise reduction techniques are applied to the experimental data in an attempt to extract any underlying deterministic dynamics. Two model systems are used to simulate the plasma dynamics. These are -the DEBS code, which models global RFP dynamics, and the dissipative trapped electron mode (DTEM) model, which models drift wave turbulence. Data from both simulations show strong indications of low,dimensional chaos and simple determinism. Experimental data were obtained from the Madison Symmetric Torus RFP and consist of a wide array of both global and local diagnostic signals. None of the signals shows any indication of low dimensional chaos or other simple determinism. Moreover, most of the analysis tools indicate the experimental system is very high dimensional with properties similar to noise. Nonlinear noise reduction is unsuccessful at extracting an underlying deterministic system

Mode dynamics and confinement in the reversed field pinch

International Nuclear Information System (INIS)

Brunsell, P.R.; Bergsaker, H.; Brzozowski, J.H.; Cecconello, M.; Drake, J.R.; Malmberg, J.-A.; Scheffel, J.; Schnack, D.D.

2001-01-01

Tearing mode dynamics and toroidal plasma flow in the RFP has been experimentally studied in the Extrap T2 device. A toroidally localised, stationary magnetic field perturbation, the 'slinky mode' is formed in nearly all discharges. There is a tendency of increased phase alignment of different toroidal Fourier modes, resulting in higher localised mode amplitudes, with higher magnetic fluctuation level. The fluctuation level increases slightly with increasing plasma current and plasma density. The toroidal plasma flow velocity and the ion temperature has been measured with Doppler spectroscopy. Both the toroidal plasma velocity and the ion temperature clearly increase with I/N. Initial, preliminary experimental results obtained very recently after a complete change of the Extrap T2 front-end system (first wall, shell, TF coil), show that an operational window with mode rotation most likely exists in the rebuilt device, in contrast to the earlier case discussed above. A numerical code DEBSP has been developed to simulate the behaviour of RFP confinement in realistic geometry, including essential transport physics. Resulting scaling laws are presented and compared with results from Extrap T2 and other RFP experiments. (author)

Chaos in reversed-field-pinch plasma simulation and experiment

International Nuclear Information System (INIS)

Watts, C.; Newman, D.E.; Sprott, J.C.

1994-01-01

We investigate the possibility that chaos and simple determinism are governing the dynamics of reversed-field-pinch (RFP) plasmas using data from both numerical simulations and experiment. A large repertoire of nonlinear-analysis techniques is used to identify low-dimensional chaos. These tools include phase portraits and Poincare sections, correlation dimension, the spectrum of Lyapunov exponents, and short-term predictability. In addition, nonlinear-noise-reduction techniques are applied to the experimental data in an attempt to extract any underlying deterministic dynamics. Two model systems are used to simulate the plasma dynamics. These are the DEBS computer code, which models global RFP dynamics, and the dissipative trapped-electron-mode model, which models drift-wave turbulence. Data from both simulations show strong indications of low-dimensional chaos and simple determinism. Experimental data were obtained from the Madison Symmetric Torus RFP and consist of a wide array of both global and local diagnostic signals. None of the signals shows any indication of low-dimensional chaos or other simple determinism. Moreover, most of the analysis tools indicate that the experimental system is very high dimensional with properties similar to noise. Nonlinear noise reduction is unsuccessful at extracting an underlying deterministic system

Confinement characteristics of the TPE reversed field pinch plasmas and effects of the boundary configuration

International Nuclear Information System (INIS)

Yagi, Y.; Maejima, Y.; Zollino, G.

2001-01-01

Confinement characteristics of the TPE series reversed field pinch (RFP) machines, TPE-1RM15, TPE-1RM20 and TPE-1RM20mod, at Electrotechnical Laboratory (ETL) are summarized. Especially data are synthesized in respect to the effects of the different boundary structures of the machines, where shell proximity and overlapped poloidal shell gaps by the multi-layered shell structure are featured. Comparison of the experimental results is shown in terms of the characteristics of magnetic fluctuations, global confinement properties in general, operation capability of the improved confinement in high pinch parameter (Q) discharges and locked mode events. Linear growth rate of the unstable modes as a function of the shell distance is numerically simulated. Understandings of RFP plasma physics have also made progress by the most recent intensive experiments on correlation studies between fast electrons and dynamo activities and measurement of the plasma and mode rotation. TPE-1RM20mod was shutdown in December 1996 and new RFP experiment has started in TPE-RX from March 1998. The new machine also succeeds the concept of the shell configuration of the TPE-1RM20. (author)

Chaos in plasma simulation and experiment

Energy Technology Data Exchange (ETDEWEB)

Watts, C. [Texas Univ., Austin, TX (United States). Fusion Research Center; Newman, D.E. [Oak Ridge National Lab., TN (United States); Sprott, J.C. [Wisconsin Univ., Madison, WI (United States). Plasma Physics Research

1993-09-01

We investigate the possibility that chaos and simple determinism are governing the dynamics of reversed field pinch (RFP) plasmas using data from both numerical simulations and experiment. A large repertoire of nonlinear analysis techniques is used to identify low dimensional chaos. These tools include phase portraits and Poincard sections, correlation dimension, the spectrum of Lyapunov exponents and short term predictability. In addition, nonlinear noise reduction techniques are applied to the experimental data in an attempt to extract any underlying deterministic dynamics. Two model systems are used to simulate the plasma dynamics. These are -the DEBS code, which models global RFP dynamics, and the dissipative trapped electron mode (DTEM) model, which models drift wave turbulence. Data from both simulations show strong indications of low,dimensional chaos and simple determinism. Experimental data were obtained from the Madison Symmetric Torus RFP and consist of a wide array of both global and local diagnostic signals. None of the signals shows any indication of low dimensional chaos or other simple determinism. Moreover, most of the analysis tools indicate the experimental system is very high dimensional with properties similar to noise. Nonlinear noise reduction is unsuccessful at extracting an underlying deterministic system.

Characteristics of a large reversed field pinch machine, TPE-RX

International Nuclear Information System (INIS)

Yagi, Y.; Shimada, T.; Hirano, Y.; Sekine, S.; Sakakita, H.; Koguchi, H.; Kiyama, S.; Maejima, Y.; Hirota, I.; Hayase, K.; Sato, Y.; Sugisaki, K.; Oyabu, I.; Hasegawa, M.; Yamane, M.; Sato, F.; Kuno, K.; Minato, T.; Kiryu, A.; Takagi, S.; Sako, K.; Kudough, F.; Urata, K.; Orita, J.; Kaguchi, H.; Sago, H.; Ue, K.

1998-01-01

Construction of a new, large reversed field pinch (RFP) machine called TPE-RX was complete at the end of 1997 as a successor of the previous TPE-1RM20 machine at the Electrotechnical Laboratory (ETL). RFP configuration has been successfully obtained in March 1998. The optimization of the operating condition and discharge cleaning of the wall are presently undergoing with the first physics experiments. This paper is the first report of TPE-RX especially on the goals, overall machine characteristics and the present status. Other papers accompanying with this one will present specific topics on the magnetic coil system and the vacuum vessel system. (author)

Characteristics of a large reversed field pinch machine, TPE-RX

Energy Technology Data Exchange (ETDEWEB)

Yagi, Y.; Shimada, T.; Hirano, Y.; Sekine, S.; Sakakita, H.; Koguchi, H.; Kiyama, S.; Maejima, Y.; Hirota, I.; Hayase, K.; Sato, Y.; Sugisaki, K. [Electrotechnical Lab., Tsukuba-shi, Ibaraki (Japan); Oyabu, I.; Hasegawa, M.; Yamane, M.; Sato, F.; Kuno, K.; Minato, T.; Kiryu, A.; Takagi, S.; Sako, K. [Mitsubishi Electric Corp. (Japan); Kudough, F.; Urata, K.; Orita, J.; Kaguchi, H.; Sago, H.; Ue, K. [Mitsubishi Heavy Industries Ltd. (Japan)

1998-07-01

Construction of a new, large reversed field pinch (RFP) machine called TPE-RX was complete at the end of 1997 as a successor of the previous TPE-1RM20 machine at the Electrotechnical Laboratory (ETL). RFP configuration has been successfully obtained in March 1998. The optimization of the operating condition and discharge cleaning of the wall are presently undergoing with the first physics experiments. This paper is the first report of TPE-RX especially on the goals, overall machine characteristics and the present status. Other papers accompanying with this one will present specific topics on the magnetic coil system and the vacuum vessel system. (author)

Magnetic and electrostatic fluctuation measurements on the ZT-40M reversed field pinch

International Nuclear Information System (INIS)

Miller, G.; Ingraham, J.C.; Munson, C.P.; Schoenberg, K.F.; Weber, P.G.; Tsui, H.Y.; Ritz, C.P.

1990-01-01

It is presently unknown whether anomalous transport in toroidal, magnetically confined plasma systems, if fluctuation induced, is dominated by electrostatic or magnetic turbulence. We are participating in a joint study of the edge plasmas of tokamak, stellarator, and RFP in an attempt to elucidate this issue. We measure magnetic and electrostatic fields using probes inserted into the edge of the ZT-40M RFP. Using the present technique, with stationary probes, these measurements can be done without damaging the probes only for low current discharges (60 kA). In this initial study, we find that both turbulent magnetic and electrostatic transport are of importance. (author) 10 refs., 2 figs., 1 tab

Post-mortem re-cloning of a transgenic red fluorescent protein dog.

Science.gov (United States)

Hong, So Gun; Koo, Ok Jae; Oh, Hyun Ju; Park, Jung Eun; Kim, Minjung; Kim, Geon-A; Park, Eun Jung; Jang, Goo; Lee, Byeong-Chun

2011-12-01

Recently, the world's first transgenic dogs were produced by somatic cell nuclear transfer. However, cellular senescence is a major limiting factor for producing more advanced transgenic dogs. To overcome this obstacle, we rejuvenated transgenic cells using a re-cloning technique. Fibroblasts from post-mortem red fluorescent protein (RFP) dog were reconstructed with in vivo matured oocytes and transferred into 10 surrogate dogs. One puppy was produced and confirmed as a re-cloned dog. Although the puppy was lost during birth, we successfully established a rejuvenated fibroblast cell line from this animal. The cell line was found to stably express RFP and is ready for additional genetic modification.

Diagnosing the pathophysiologic mechanisms of nocturnal polyuria.

Science.gov (United States)

Goessaert, An-Sofie; Krott, Louise; Hoebeke, Piet; Vande Walle, Johan; Everaert, Karel

2015-02-01

Diagnosis of nocturnal polyuria (NP) is based on a bladder diary. Addition of a renal function profile (RFP) for analysis of concentrating and solute-conserving capacity allows differentiation of NP pathophysiology and could facilitate individualized treatment. To map circadian rhythms of water and solute diuresis by comparing participants with and without NP. This prospective observational study was carried out in Ghent University Hospital between 2011 and 2013. Participants with and without NP completed a 72-h bladder dairy. RFP, free water clearance (FWC), and creatinine, solute, sodium, and urea clearance were measured for all participants. The study participants were divided into those with (n=77) and those without (n=35) NP. The mean age was 57 yr (SD 16 yr) and 41% of the participants were female. Compared to participants without NP, the NP group exhibited a higher diuresis rate throughout the night (p=0.015); higher FWC (p=0.013) and lower osmolality (p=0.030) at the start of the night; and persistently higher sodium clearance during the night (p<0.001). The pathophysiologic mechanism of NP was identified as water diuresis alone in 22%, sodium diuresis alone in 19%, and a combination of water and sodium diuresis in 47% of the NP group. RFP measurement in first-line NP screening to discriminate between water and solute diuresis as pathophysiologic mechanisms complements the bladder diary and could facilitate optimal individualized treatment of patients with NP. We evaluated eight urine samples collected over 24h to detect the underlying problem in NP. We found that NP can be attributed to water or sodium diuresis or a combination of both. This urinalysis can be used to adapt treatment according to the underlying mechanism in patients with bothersome consequences of NP, such as nocturia and urinary incontinence. Copyright © 2014 European Association of Urology. Published by Elsevier B.V. All rights reserved.

Determination of pKa constants of hypericin in aqueous solution of the anti-allergic hydrotropic drug Cromolyn disodium salt

Science.gov (United States)

Keša, Peter; Antalík, Marián

2017-05-01

In this work we established three from altogether six proton dissociation constants (pKa) of hydroxyl groups of hypericin in its monomeric form. The monomeric state of hypericin (5.0 × 10-6 mol·L-1) in aqueous solution was stabilised by the presence of hydrotropic drug Cromolyn disodium salt (6.0 × 10-2 mol·L-1). Data show that one acid-base transition occurs with the pKa of 7.8 and the other two are characterised by the apparent single pKa of 11.5. The spectral changes of hypericin above pH 13 indicate that the last two hydroxyls are deporotonized at this high pH values.

Group contribution methodology based on the statistical associating fluid theory for heteronuclear molecules formed from Mie segments.

Science.gov (United States)

Papaioannou, Vasileios; Lafitte, Thomas; Avendaño, Carlos; Adjiman, Claire S; Jackson, George; Müller, Erich A; Galindo, Amparo

2014-02-07

A generalization of the recent version of the statistical associating fluid theory for variable range Mie potentials [Lafitte et al., J. Chem. Phys. 139, 154504 (2013)] is formulated within the framework of a group contribution approach (SAFT-γ Mie). Molecules are represented as comprising distinct functional (chemical) groups based on a fused heteronuclear molecular model, where the interactions between segments are described with the Mie (generalized Lennard-Jonesium) potential of variable attractive and repulsive range. A key feature of the new theory is the accurate description of the monomeric group-group interactions by application of a high-temperature perturbation expansion up to third order. The capabilities of the SAFT-γ Mie approach are exemplified by studying the thermodynamic properties of two chemical families, the n-alkanes and the n-alkyl esters, by developing parameters for the methyl, methylene, and carboxylate functional groups (CH3, CH2, and COO). The approach is shown to describe accurately the fluid-phase behavior of the compounds considered with absolute average deviations of 1.20% and 0.42% for the vapor pressure and saturated liquid density, respectively, which represents a clear improvement over other existing SAFT-based group contribution approaches. The use of Mie potentials to describe the group-group interaction is shown to allow accurate simultaneous descriptions of the fluid-phase behavior and second-order thermodynamic derivative properties of the pure fluids based on a single set of group parameters. Furthermore, the application of the perturbation expansion to third order for the description of the reference monomeric fluid improves the predictions of the theory for the fluid-phase behavior of pure components in the near-critical region. The predictive capabilities of the approach stem from its formulation within a group-contribution formalism: predictions of the fluid-phase behavior and thermodynamic derivative properties of

Viral Cre-LoxP tools aid genome engineering in mammalian cells.

Science.gov (United States)

Sengupta, Ranjita; Mendenhall, Amy; Sarkar, Nandita; Mukherjee, Chandreyee; Afshari, Amirali; Huang, Joseph; Lu, Biao

2017-01-01

Targeted nucleases have transformed genome editing technology, providing more efficient methods to make targeted changes in mammalian genome. In parallel, there is an increasing demand of Cre-LoxP technology for complex genome manipulation such as large deletion, addition, gene fusion and conditional removal of gene sequences at the target site. However, an efficient and easy-to-use Cre-recombinase delivery system remains lacking. We designed and constructed two sets of expression vectors for Cre-recombinase using two highly efficient viral systems, the integrative lentivirus and non-integrative adeno associated virus. We demonstrate the effectiveness of those methods in Cre-delivery into stably-engineered HEK293 cells harboring LoxP-floxed red fluorescent protein (RFP) and puromycin (Puro) resistant reporters. The delivered Cre recombinase effectively excised the floxed RFP-Puro either directly or conditionally, therefore validating the function of these molecular tools. Given the convenient options of two selections markers, these viral-based systems offer a robust and easy-to-use tool for advanced genome editing, expanding complicated genome engineering to a variety of cell types and conditions. We have developed and functionally validated two viral-based Cre-recombinase delivery systems for efficient genome manipulation in various mammalian cells. The ease of gene delivery with the built-in reporters and inducible element enables live cell monitoring, drug selection and temporal knockout, broadening applications of genome editing.

Study of Opacity Effects on Emission Lines at EXTRAP T2R RFP

Science.gov (United States)

Stancalie, Viorica; Rachlew, Elisabeth

We have investigated the influence of opacity on hydrogen (H-α and Ly-β) and Li-like oxygen emission lines from the EXTRAP T2R reversed field pinch. We used the Atomic Data Analysis System (AzDAS) based on the escape factor approximation for radiative transfer to calculate metastable and excited population densities via a collisional-radiative model. Population escape factor, emergent escape factor and modified line profiles are plotted vs. optical depth. The simulated emission line ratios in the density/temperature plane are in good agreement with experimental data for electron density and temperature measurements.

Characterisation of Vranec, Cabernet sauvignon and Merlot wines based on their chromatic and anthocyanin profiles

OpenAIRE

Dimitrovska Maja; Tomovska Elena; Bocevska Mirjana

2013-01-01

Wines of three different grape varieties, Vranec, Cabernet Sauvignon and Merlot were examined for their characterisation in terms of anthocyanin and chromatic profiles, total polyphenols and antioxidant potential. Total, monomeric, polymeric and copigmented anthocyanins were determined by spectrophotometry and the individual anthocyanin compounds were quantified using HPLC-DAD. Chromatic profile was evaluated according to colour density, hue, % red, % blue, % yellow and brilliance (% dA...

Soft X-ray diagnostic at the EXTRAP-T1 experiment

International Nuclear Information System (INIS)

Voronin, A.; Welander, A.

1992-12-01

Results of investigating the soft x-ray wavelength region of reversed field pinch (RFP) discharges at the EXTRAP-T1 device are presented. A non-dispersive technique based on the multiple-filters method has been developed to measure the electron temperature and plasma instabilities. Time resolved local temperature measurements of 100-300 eV near the torus axis were carried out. The electron temperature dependence on plasma current and conditions of the operation of the device has been experimentally established. (authors)

Ccr2 deletion dissociates cavity size and tau pathology after mild traumatic brain injury.

Science.gov (United States)

Gyoneva, Stefka; Kim, Daniel; Katsumoto, Atsuko; Kokiko-Cochran, O Nicole; Lamb, Bruce T; Ransohoff, Richard M

2015-12-03

Millions of people experience traumatic brain injury (TBI) as a result of falls, car accidents, sports injury, and blast. TBI has been associated with the development of neurodegenerative conditions such as Alzheimer's disease (AD) and chronic traumatic encephalopathy (CTE). In the initial hours and days, the pathology of TBI comprises neuronal injury, breakdown of the blood-brain barrier, and inflammation. At the cellular level, the inflammatory reaction consists of responses by brain-resident microglia, astrocytes, and vascular elements as well as infiltration of peripheral cells. After TBI, signaling by chemokine (C-C motif) ligand 2 (CCL2) to the chemokine (C-C motif) receptor 2 (CCR2) is a key regulator of brain infiltration by monocytes. We utilized mice with one or both copies of Ccr2 disrupted by red fluorescent protein (RFP, Ccr2 (RFP/+) and Ccr2 (RFP/RFP) ). We subjected these mice to the mild lateral fluid percussion model of TBI and examined several pathological outcomes 3 days later in order to determine the effects of altered monocyte entry into the brain. Ccr2 deletion reduced monocyte infiltration, diminished lesion cavity volume, and lessened axonal damage after mild TBI, but the microglial reaction to the lesion was not affected. We further examined phosphorylation of the microtubule-associated protein tau, which aggregates in brains of people with TBI, AD, and CTE. Surprisingly, Ccr2 deletion was associated with increased tau mislocalization to the cell body in the cortex and hippocampus by tissue staining and increased levels of phosphorylated tau in the hippocampus by Western blot. Disruption of CCR2 enhanced tau pathology and reduced cavity volume in the context of TBI. The data reveal a complex role for CCR2(+) monocytes in TBI, as monitored by cavity volume, axonal damage, and tau phosphorylation.

Update of ZTH physics and design issues and physics goals of ZTH and role in the Fusion Program

International Nuclear Information System (INIS)

DiMarco, J.N.

1990-01-01

The ZTH construction program is scheduled for completion in December of 1992. Some design features are still amenable to changes directed by new physics or computational information. Numerical results for the ZTH tapered poloidal field gap show that a relatively simple linear taper results in substantial reduction in field error. This design is simpler to manufacture compared with the compound curve predicted by analytical calculations. Also, ongoing analysis of ZT-40M data indicates that the fluctuation levels of magnetic fields and x-rays at high θ (1.7), can be reduced to the fluctuation levels at ''standard'' operational θ (1.4) by changing the winding configuration of the toroidal field (TF) coils. The effect is thought to depend on the shell-like action of the TF coils when they consist of many turns in parallel. Magnetic-field effects, such as field errors, at the unshielded toroidal-field butt-joint gap in the shell can be reduced by this effective external shell. Design implications for installing a second low-current TF coil on ZTH are presented. ZTH has the capability of operating at 4 MA with the addition of power supplies. The projected parameters of ZTH are discussed in the context of various conceptual design. The sensitivity of the conceptual design to the physics results from machines like RFX and ZTH is reviewed. It is shown that the 4 MA ZTH physics results will make a significant advance for the RFP program toward the programmatic RFP fusion goals. Finally, the influence of RFP and tokamak physics on the conceptual design of high mass-power-density (MPD) fusion reactors is investigated through the TITAN and ARIES studies. The confinement characteristics required of these conceptual designs are compared; physics issues are identified that are required to bring the MPD and fusion-power-core of the tokamak designs into coincidence with the RFP designs

Identification of choriogenin cis-regulatory elements and production of estrogen-inducible, liver-specific transgenic Medaka.

Science.gov (United States)

Ueno, Tetsuro; Yasumasu, Shigeki; Hayashi, Shinji; Iuchi, Ichiro

2004-07-01

Choriogenins (chg-H, chg-L) are precursor proteins of egg envelope of medaka and synthesized in the spawning female liver in response to estrogen. We linked a gene construct chg-L1.5 kb/GFP (a 1.5 kb 5'-upstream region of the chg-L gene fused with a green fluorescence protein (GFP) gene) to another construct emgb/RFP (a cis-regulatory region of embryonic globin gene fused with an RFP gene), injected the double fusion gene construct into 1- or 2-cell-stage embryos, and selected embryos expressing the RFP in erythroid cells. From the embryos, we established two lines of chg-L1.5 kb/GFP-emgb/RFP-transgenic medaka. The 3-month-old spawning females and estradiol-17beta (E2)-exposed males displayed the liver-specific GFP expression. The E2-dependent GFP expression was detected in the differentiating liver of the stage 37-38 embryos. In addition, RT-PCR and whole-mount in situ hybridization showed that the E2-dependent chg expression was found in the liver of the stage 34 embryos of wild medaka, suggesting that such E2-dependency is achieved shortly after differentiation of the liver. Analysis using serial deletion mutants fused with GFP showed that the region -426 to -284 of the chg-L gene or the region -364 to -265 of the chg-H gene had the ability to promote the E2-dependent liver-specific GFP expression of its downstream gene. Further analyses suggested that an estrogen response element (ERE) at -309, an ERE half-site at -330 and a binding site for C/EBP at -363 of the chg-L gene played important roles in its downstream chg-L gene expression. In addition, this transgenic medaka may be useful as one of the test animals for detecting environmental estrogenic steroids.

Palm oil based polymer materials obtained by ROMP: study by low field NMR; Materiais polimericos obtidos via ROMP a partir de oleo de palma: estudo por RMN de baixo campo

Energy Technology Data Exchange (ETDEWEB)

Fernandes, Henrique; Azevedo, Eduardo R. de; Lima-Neto, Benedito S., E-mail: benedito@iqsc.usp.br [Universidade de Sao Paulo (USP), Sao Carlos, SP (Brazil)

2015-07-01

Aiming to study and develop new materials synthesized from sustainable sources, several polymers were prepared using in its monomeric composition, different amounts of NPO (Norbornenyl Palm Oil) monomer. This monomer was developed based on a vegetable oil rather produced in northern Brazil, the Palm Oil. Since this oil have a low content of unsaturation, its use in developing new monomer for ROMP (Ring-Opening Metathesis Polymerization) is not exploited. In this regard, polymeric materials were obtained using the NOP and both the reaction process and the resulting products were analyzed by Nuclear Magnetic Resonance in the time domain (TD-NMR) at low magnetic field. (author)

Palm oil based polymer materials obtained by romp: study by low field NMR; Materiais polimericos obtidos via ROMP a partir de oleo de palma: estudo por RMN de baixo campo

Energy Technology Data Exchange (ETDEWEB)

Fernandes, Henrique; Lima-Neto, Benedito S., E-mail: benedito@iqsc.usp.br [Universidade de Sao Paulo (USP), Sao Carlos, SP (Brazil). Instituto de Quimica de; Azevedo, Eduardo R. de [Universidade de Sao Paulo (USP), Sao Carlos, SP (Brazil). Instituto de Fisica

2013-07-01

Aiming to study and develop new materials synthesized from sustainable sources, several polymers were prepared using in its monomeric composition, different amounts of NPO (Norbornenyl Palm Oil) monomer. This monomer was developed based on a vegetable oil rather produced in northern Brazil, the Palm Oil. Since this oil have a low content of unsaturation, its use in developing new monomer for ROMP (Ring-Opening Metathesis Polymerization) is not exploited. In this regard, polymeric materials were obtained using the NOP and both the reaction process and the resulting products were analyzed by Nuclear Magnetic Resonance in the time domain (TD-NMR) at low magnetic field. (author)

Confinement dynamics in the ZT-40M RFP

International Nuclear Information System (INIS)

Weber, P.G.; Ingraham, J.C.; Miller, G.; Munson, C.P.; Phillips, J.A.; Pickrell, M.M.; Schoenberg, K.F.; Wurden, G.A.

1989-01-01

Typically one-third of the input power is accounted for by radiation as measured by bolometers. This radiated power fraction has been varied from 0.15 to 0.95 in impurity injection experiments, with the global β and τ E values maintained to within a factor two. This implies that the non-radiative confinement varied by a factor five or more while the total confinement remained roughly the same

Characterization of the Sweet Taste Receptor Tas1r2 from an Old World Monkey Species Rhesus Monkey and Species-Dependent Activation of the Monomeric Receptor by an Intense Sweetener Perillartine.

Directory of Open Access Journals (Sweden)

Chenggu Cai

Full Text Available Sweet state is a basic physiological sensation of humans and other mammals which is mediated by the broadly acting sweet taste receptor-the heterodimer of Tas1r2 (taste receptor type 1 member 2 and Tas1r3 (taste receptor type 1 member 3. Various sweeteners interact with either Tas1r2 or Tas1r3 and then activate the receptor. In this study, we cloned, expressed and functionally characterized the taste receptor Tas1r2 from a species of Old World monkeys, the rhesus monkey. Paired with the human TAS1R3, it was shown that the rhesus monkey Tas1r2 could respond to natural sugars, amino acids and their derivates. Furthermore, similar to human TAS1R2, rhesus monkey Tas1r2 could respond to artificial sweeteners and sweet-tasting proteins. However, the responses induced by rhesus monkey Tas1r2 could not be inhibited by the sweet inhibitor amiloride. Moreover, we found a species-dependent activation of the Tas1r2 monomeric receptors of human, rhesus monkey and squirrel monkey but not mouse by an intense sweetener perillartine. Molecular modeling and sequence analysis indicate that the receptor has the conserved domains and ligand-specific interactive residues, which have been identified in the characterized sweet taste receptors up to now. This is the first report of the functional characterization of sweet taste receptors from an Old World monkey species.

Characterization of the Sweet Taste Receptor Tas1r2 from an Old World Monkey Species Rhesus Monkey and Species-Dependent Activation of the Monomeric Receptor by an Intense Sweetener Perillartine.

Science.gov (United States)

Cai, Chenggu; Jiang, Hua; Li, Lei; Liu, Tianming; Song, Xuejie; Liu, Bo

2016-01-01

Sweet state is a basic physiological sensation of humans and other mammals which is mediated by the broadly acting sweet taste receptor-the heterodimer of Tas1r2 (taste receptor type 1 member 2) and Tas1r3 (taste receptor type 1 member 3). Various sweeteners interact with either Tas1r2 or Tas1r3 and then activate the receptor. In this study, we cloned, expressed and functionally characterized the taste receptor Tas1r2 from a species of Old World monkeys, the rhesus monkey. Paired with the human TAS1R3, it was shown that the rhesus monkey Tas1r2 could respond to natural sugars, amino acids and their derivates. Furthermore, similar to human TAS1R2, rhesus monkey Tas1r2 could respond to artificial sweeteners and sweet-tasting proteins. However, the responses induced by rhesus monkey Tas1r2 could not be inhibited by the sweet inhibitor amiloride. Moreover, we found a species-dependent activation of the Tas1r2 monomeric receptors of human, rhesus monkey and squirrel monkey but not mouse by an intense sweetener perillartine. Molecular modeling and sequence analysis indicate that the receptor has the conserved domains and ligand-specific interactive residues, which have been identified in the characterized sweet taste receptors up to now. This is the first report of the functional characterization of sweet taste receptors from an Old World monkey species.

New insights into MHD dynamics of magnetically confined plasmas from experiments in RFX

International Nuclear Information System (INIS)

Martin, P.; Martini, S.; Antoni, V.

2001-01-01

The experimental and theoretical activity performed in the RFX experiment has allowed a deeper insight into the MHD properties of the RFP configuration. A set of successful experiments has demonstrated the possibility of influencing both the amplitude and the spectrum of the magnetic fluctuations which characterise the RFP configuration. A new regime (QSH states) where the dynamo mechanism works in a nearly laminar way and a helical core plasma is produced has been investigated. With these studies a reduction of the magnetic chaos has been obtained. The continuos rotation of wall locked resistive tearing modes has been obtained by an m=0 rotating perturbation. This perturbation induces rotation of m=1 non-linearly coupled modes. (author)

Electrical design of a high current density air-core reversed-field pinch ''ZTP''

International Nuclear Information System (INIS)

Reass, W.A.; Cribble, R.F.; Melton, J.G.

1983-01-01

This paper describes the electrical design of a small, high current density (10 MA/m 2 ) toroidal reversed-field Z-Pinch (RFP) presently being constructed at Los Alamos. Special purpose magnetic field programs were used to calculate self and mutual inductances for the poloidal field windings. The network analysis program MINI-SCEPTRE was then used to predict plasma current, including the interaction between toroidal and poloidal field circuits, as described by the Bessel function model for RFP's. Using these programs, coil geometry was obtained for minimal field errors and the pulse power systems were optimized to minimize equilibrium control power. Results of computer modeling and implementation of the electrical circuits are presented

Electrical design of a high current density air-core reversed-field pinch ZTP

International Nuclear Information System (INIS)

Reass, W.A.; Melton, J.G.; Gribble, R.F.

1983-01-01

This paper describes the electrical design of a small, high current density (10 MA/m 2 ) toroidal reversed-field Z-Pinch (RFP) presently being constructed at Los Alamos. Special purpose magnetic field programs were used to calculate self and mutual inductances for the poloidal field windings. The network analysis program MINI-SCEPTRE was then used to predict plasma current, including the interaction between toroidal and poloidal field circuits, as described by the Bessel function model for RFP's. Using these programs, coil geometry was obtained for minimal field errors and the pulse power systems were optimized to minimize equilibrium control power. Results of computer modeling and implementation of the electrical circuits are presented

Ohm close-quote s law for plasmas in reversed field pinch configuration

International Nuclear Information System (INIS)

Martines, E.; Vallone, F.

1997-01-01

An analytical relationship between current density and applied electric field in reversed field pinch (RFP) plasmas has been derived in the framework of the kinetic dynamo theory, that is assuming a radial field-aligned momentum transport caused by the magnetic field stochasticity. This Ohm close-quote s law yields current density profiles with a poloidal current density at the edge which can sustain the magnetic field configuration against resistive diffusion. The dependence of the loop voltage on plasma current and other plasma parameters for RFP experiments has been obtained. The results of the theoretical work have been compared with experimental data from the RFX experiment, and a good agreement has been found. copyright 1997 The American Physical Society

The use of the case study method in radiation worker continuing training

International Nuclear Information System (INIS)

Stevens, R.D.

1990-01-01

Typical methods of continuing training are often viewed by employees as boring, redundant and unnecessary. It is hoped that the operating experience lesson in the required course, Radiation Worker Requalification, will be well received by employees because actual RFP events will be presented as case studies. The interactive learning atmosphere created by the case study method stimulates discussion, develops analytical abilities, and motivates employees to use lessons learned in the workplace. This problem solving approach to continuing training incorporates cause and effect analysis, a technique which is also used at RFP to investigate events. A method of designing the operating experience lesson in the Radiation Worker Requalification course is described in this paper. 7 refs., 2 figs

Isotopic ratio method for determining uranium contamination

International Nuclear Information System (INIS)

Miles, R.E.; Sieben, A.K.

1994-01-01

The presence of high concentrations of uranium in the subsurface can be attributed either to contamination from uranium processing activities or to naturally occurring uranium. A mathematical method has been employed to evaluate the isotope ratios from subsurface soils at the Rocky Flats Nuclear Weapons Plant (RFP) and demonstrates conclusively that the soil contains uranium from a natural source and has not been contaminated with enriched uranium resulting from RFP releases. This paper describes the method used in this determination which has widespread application in site characterizations and can be adapted to other radioisotopes used in manufacturing industries. The determination of radioisotope source can lead to a reduction of the remediation effort

Recovery of transuranics from process residues

International Nuclear Information System (INIS)

Gray, J.H.; Gray, L.W.

1987-01-01

Process residues are generated at both the Rocky Flats Plant (RFP) and the Savannah River Plant (SRP) during aqueous chemical and pyrochemical operations. Frequently, process operations will result in either impure products or produce residues sufficiently contaminated with transuranics to be nondiscardable as waste. Purification and recovery flowsheets for process residues have been developed to generate solutions compatible with subsequent Purex operations and either solid or liquid waste suitable for disposal. The ''scrub alloy'' and the ''anode heel alloy'' are examples of materials generated at RFP which have been processed at SRP using the developed recovery flowsheets. Examples of process residues being generated at SRP for which flowsheets are under development include LECO crucibles and alpha-contaminated hydraulic oil

Eddy current analysis in fusion devices

International Nuclear Information System (INIS)

Turner, L.R.

1988-06-01

In magnetic fusion devices, particularly tokamaks and reversed field pinch (RFP) experiments, time-varying magnetic fields are in intimate contact with electrically conducting components of the device. Induced currents, fields, forces, and torques result. This note reviews the analysis of eddy current effects in the following systems: Interaction of a tokamak plasma with the eddy currents in the first wall, blanket, and shield (FWBS) systems; Eddy currents in a complex but two-dimensional vacuum vessel, as in TFTR, JET, and JT-60; Eddy currents in the FWBS system of a tokamak reactor, such as NET, FER, or ITER; and Eddy currents in a RFP shell. The cited studies are chosen to be illustrative, rather than exhaustive. 42 refs

Effect of temperature on the radioiodination of human growth hormone

International Nuclear Information System (INIS)

Mohammed-Ali, S.A.; Salacinski, P.R.; Landon, J.

1981-01-01

Studies have been undertaken to assess the effect of altering the temperature at which human growth hormone is radioiodinated on the incorporation of 125 I and the immunoreactivity and stability of the labelled hormone. Employing highly purified monomeric hormone it proved possible, by the iodogen procedure, to prepare a labelled product of high specific activity irrespective of temperature. However, in radioiodinations performed at ambient temperature (20 to 25 degrees) significant amounts of the labelled hormone were in an aggregated form which was less immunoreactive than the 125 I-labelled monomeric hormone. Such aggregation was largely prevented by radioiodinating at low temperature (0 to 4 degrees) and even the large monomeric peak was more immunoreactive (about 95% bound in antibody excess) than the monomeric peak from iodinations performed at room temperature

Atomic Force Microscopy Study of Protein–Protein Interactions in the Cytochrome CYP11A1 (P450scc-Containing Steroid Hydroxylase System

Directory of Open Access Journals (Sweden)

Zöllner A

2011-01-01

Full Text Available Abstract Atomic force microscopy (AFM and photon correlation spectroscopy (PCS were used for monitoring of the procedure for cytochrome CYP11A1 monomerization in solution without phospholipids. It was shown that the incubation of 100 μM CYP11A1 with 12% Emulgen 913 in 50 mM KP, pH 7.4, for 10 min at T = 22°C leads to dissociation of hemoprotein aggregates to monomers with the monomerization degree of (82 ± 4%. Following the monomerization procedure, CYP11A1 remained functionally active. AFM was employed to detect and visualize the isolated proteins as well as complexes formed between the components of the cytochrome CYP11A1-dependent steroid hydroxylase system. Both Ad and AdR were present in solution as monomers. The typical heights of the monomeric AdR, Ad and CYP11A1 images were measured by AFM and were found to correspond to the sizes 1.6 ± 0.2 nm, 1.0 ± 0.2 nm and 1.8 ± 0.2 nm, respectively. The binary Ad/AdR and AdR/CYP11A1mon complexes with the heights 2.2 ± 0.2 nm and 2.8 ± 0.2 nm, respectively, were registered by use of AFM. The Ad/CYP11A1mon complex formation reaction was kinetically characterized based on optical biosensor data. In addition, the ternary AdR/Ad/CYP11A1 complexes with a typical height of 4 ± 1 nm were AFM registered.

Turbulence, transport, and zonal flows in the Madison symmetric torus reversed-field pinch

Science.gov (United States)

Williams, Z. R.; Pueschel, M. J.; Terry, P. W.; Hauff, T.

2017-12-01

The robustness and the effect of zonal flows in trapped electron mode (TEM) turbulence and Ion Temperature Gradient (ITG) turbulence in the reversed-field pinch (RFP) are investigated from numerical solutions of the gyrokinetic equations with and without magnetic external perturbations introduced to model tearing modes. For simulations without external magnetic field perturbations, zonal flows produce a much larger reduction of transport for the density-gradient-driven TEM turbulence than they do for the ITG turbulence. Zonal flows are studied in detail to understand the nature of their strong excitation in the RFP and to gain insight into the key differences between the TEM- and ITG-driven regimes. The zonal flow residuals are significantly larger in the RFP than in tokamak geometry due to the low safety factor. Collisionality is seen to play a significant role in the TEM zonal flow regulation through the different responses of the linear growth rate and the size of the Dimits shift to collisionality, while affecting the ITG only minimally. A secondary instability analysis reveals that the TEM turbulence drives zonal flows at a rate that is twice that of the ITG turbulence. In addition to interfering with zonal flows, the magnetic perturbations are found to obviate an energy scaling relation for fast particles.

High-flux first-wall design for a small reversed-field pinch reactor

International Nuclear Information System (INIS)

Cort, G.E.; Graham, A.L.; Christensen, K.E.

1982-01-01

To achieve the goal of a commercially economical fusion power reactor, small physical size and high power density should be combined with simplicity (minimized use of high-technology systems). The Reversed-Field Pinch (RFP) is a magnetic confinement device that promises to meet these requirements with power densities comparable to those in existing fission power plants. To establish feasibility of such an RFP reactor, a practical design for a first wall capable of withstanding high levels of cyclic neutron wall loadings is needed. Associated with the neutron flux in the proposed RFP reactor is a time-averaged heat flux of 4.5 MW/m 2 with a conservatively estimated transient peak approximately twice the average value. We present the design for a modular first wall made from a high-strength copper alloy that will meet these requirements of cyclic thermal loading. The heat removal from the wall is by subcooled water flowing in straight tubes at high linear velocities. We combined a thermal analysis with a structural fatigue analysis to design the heat transfer module to last 10 6 cycles or one year at 80% duty for a 26-s power cycle. This fatigue life is compatible with a radiation damage life of 14 MW/yr/m 2

Commercial Decommissioning at DOE's Rocky Flats

International Nuclear Information System (INIS)

Freiboth, C.; Sandlin, N.; Schubert, A.; Hansen, S.

2002-01-01

Due in large part to the number of nuclear facilities that make up the DOE complex, DOE-EM work has historically been paperwork intensive and driven by extensive regulations. Requirements for non-nuclear facilities are often grouped with those of nuclear facilities, driving up costs. Kaiser-Hill was interested in applying a commercial model to demolition of these facilities and wanted to apply necessary and sufficient standards to the work activities, but avoid applying unnecessary requirements. Faced with demolishing hundreds of uncontaminated or non-radiologically contaminated facilities, Kaiser-Hill has developed a subcontracting strategy to drastically reduce the cost of demolishing these facilities at Rocky Flats. Aiming to tailor the demolition approach of such facilities to more closely follow commercial practices, Kaiser-Hill recently released a Request for Proposals (RFP) for the demolition of the site's former central administration facility. The RFP significantly reduced requirements for compliance with specific DOE directives. Instead, the RFP required subcontractors to comply with health and safety requirements commonly found in the demolition of similar facilities in a commercial setting. This resulted in a number of bids from companies who have normally not bid on DOE work previously and at a reduced cost over previous approaches. This paper will discuss the details of this subcontracting strategy

Rett Syndrome Mutant Neural Cells Lacks MeCP2 Immunoreactive Bands.

Directory of Open Access Journals (Sweden)

Carlos Bueno

Full Text Available Dysfunctions of MeCP2 protein lead to various neurological disorders such as Rett syndrome and Autism. The exact functions of MeCP2 protein is still far from clear. At a molecular level, there exist contradictory data. MeCP2 protein is considered a single immunoreactive band around 75 kDa by western-blot analysis but several reports have revealed the existence of multiple MeCP2 immunoreactive bands above and below the level where MeCP2 is expected. MeCP2 immunoreactive bands have been interpreted in different ways. Some researchers suggest that multiple MeCP2 immunoreactive bands are unidentified proteins that cross-react with the MeCP2 antibody or degradation product of MeCP2, while others suggest that MeCP2 post-transcriptional processing generates multiple molecular forms linked to cell signaling, but so far they have not been properly analyzed in relation to Rett syndrome experimental models. The purpose of this study is to advance understanding of multiple MeCP2 immunoreactive bands in control neural cells and p.T158M MeCP2e1 mutant cells. We have generated stable wild-type and p.T158M MeCP2e1-RFP mutant expressing cells. Application of N- and C- terminal MeCP2 antibodies, and also, RFP antibody minimized concerns about nonspecific cross-reactivity, since they react with the same antigen at different epitopes. We report the existence of multiple MeCP2 immunoreactive bands in control cells, stable wild-type and p.T158M MeCP2e1-RFP mutant expressing cells. Also, MeCP2 immunoreactive bands differences were found between wild-type and p.T158M MeCP2e1-RFP mutant expressing cells. Slower migration phosphorylated band around 70kDa disappeared in p.T158M MeCP2e1-RFP mutant expressing cells. These data suggest that threonine 158 could represent an important phosphorylation site potentially involved in protein function. Our results clearly indicate that MeCP2 antibodies have no cross-reactivity with similar epitopes on others proteins, supporting the

EFECTOS HETERÓTICOS Y HABILIDAD COMBINATORIA PARA EL RENDIMIENTO POR PLANTA EN Cucurbita moschata DUCH. Ex Poir HETEROTIC EFFECTS AND COMBINING ABILITY FOR YIELD PER PLANT IN Cucurbita moschata DUCH. Ex Poir

Directory of Open Access Journals (Sweden)

Miguel Mariano Espitia Camacho

2006-06-01

Full Text Available El estudio tuvo como objetivos estimar los efectos heteróticos y de habilidad combinatoria (HCG y específica (HCE, para el rendimiento/planta (RFP, en dos dialelos [uno entre cinco variedades (VAR y el otro entre cinco líneas endogámicas S1 (LS1, derivadas de éstas], mediante las metodologías propuestas por Gardner y Eberhart (GyE: 1966: Análisis II y Hallauer y Miranda (HyM:1981. Los genotipos se evaluaron en Candelaria (Valle del Cauca, Colombia, en un diseño de bloques completos al azar con cinco repeticiones. Las diferencias entre genotipos en el dialelo de LS1 por GyE, fueron explicadas por los efectos de variedades (v j: p jj’: p : p The purpose of this study was to estimate the heterotic effects, general combining ability (GCA, and specific (SCA for fruit yield per plant (RFP in two diallelic [one among five cross pollinated varieties (VAR and the other among five self-pollinated lines S1 (LS1, derived from these], according to the methodologies designed by Gardner y Eberhart (GyE: 1966: Analysis II and Hallauer and Miranda (HyM: 1981 using a complete randomized design with five replicates in Candelaria (Valle del Cauca, Colombia. The differences between genotypes observed in the diallelic of LS1 by GyE, were explained by the effects of varieties (v j: p (h jj’: p : p < 0,01 and specific heterosis (Sjj’: p < 0,05 explained the differences for h jj in the diallelic of LS1. According to the methodology of HyM, the additive (GCA: p < 0,01 and non additive genic action (SCA: p < 0,05 controlled the expression of RFP in the diallele of LS1 with greater importance of the GCA. In the diallele of VAR, only the GCA (p < 0,01 explained such control. The genotypes Bolo Verde (RFP = 17,8 kg/plant; v j = 3,34**; g i = 1,73* and Soler (PFP = 17 kg/plant; v j = 2,89*; g i = 175* are the recommended genotypes to exploit the additives effects and the hybrid LA34 x LSO (RFP = 26 kg/plant; Sjj’= 3,55** to exploit the additive and non

40 CFR 51.491 - Definitions.

Science.gov (United States)

2010-07-01

... different users. RFP baseline means the total of actual volatile organic compounds or nitrogen oxides..., industrial equipment, construction vehicles, off-road motorcycles, and marine vessels). National ambient air...

75 FR 3243 - National Cancer Institute; Notice of Meeting

Science.gov (United States)

2010-01-20

...); and Budget Presentations; Reports of Special Initiatives; RFA and RFP Concept Reviews; and Scientific... campus. All visitor vehicles, including taxicabs, hotel, and airport shuttles will be inspected before...

Polymerization of amino acids containing nucleotide bases

Science.gov (United States)

Ben Cheikh, Azzouz; Orgel, Leslie E.

1990-01-01

The nucleoamino acids 1-(3'-amino,3'-carboxypropyl)uracil (3) and 9-(3'-amino,3'-carboxypropyl)adenine (4) have been prepared as (L)-en-antiomers and as racemic mixtures. When 3 or 4 is suspended in water and treated with N,N'-carbon-yldiimidazole, peptides are formed in good yield. The products formed from the (L)-enantiomers are hydrolyzed to the monomeric amino acids by pronase. Attempts to improve the efficiency of these oligomerizations by including a polyuridylate template in the reaction mixture were not successful. Similarly, oligomers derived from the (L)-enantiomer of 3 did not act as templates to facilitate the oligomerization of 4.

75 FR 26761 - National Cancer Institute; Notice of Meeting

Science.gov (United States)

2010-05-12

... Group(s); and Budget Presentations; Reports of Special Initiatives; RFA and RFP Concept Reviews; and... campus. All visitor vehicles, including taxicabs, hotel, and airport shuttles will be inspected before...

78 FR 60887 - National Cancer Institute; Notice of Meeting

Science.gov (United States)

2013-10-02

...); and Budget Presentations, Reports of Special Initiatives; RFA and RFP Concept Reviews; and Scientific..., including taxicabs, hotel, and airport shuttles will be inspected before being allowed on campus. Visitors...

Preparation of gold nanoparticles in the presence of citric acid-based dendrimers containing periphery hydroxyl groups

International Nuclear Information System (INIS)

Namazi, Hassan; Fard, Ahmad Mohammad Pour

2011-01-01

Highlights: → The most advantage of citric acid-based dendrimers is their novelty from monomeric point of view and their simple preparation method. → The size and also size distribution of Au nanoparticles can be controlled through the choice of the dendrimer generation. → Here, we report the preparation of the stable, isolated and uniform Au nanoparticles with using a simple method in water media.→ It was observed that the size of Au nanoparticles is increased with increasing the generation of dendrimer. - Abstract: In this work, Au nanoparticles were produced with reduction of HAuCl 4 using NaBH 4 in the presence of different generations of citric acid-based dendrimers. The greater water solubility of the newly prepared dendrimers motivated us for the preparation of Au nanoparticles in water media. Therefore, the stable, isolated and uniform type Au nanoparticles were prepared through simple process in water. UV-Vis absorption, high-resolution transmission electronic microscopy (HRTEM), electron diffraction (ED) and energy dispersive X-ray (EDX) methods were used to investigate the morphology and structure determination of the obtained gold nanoparticles.

Pretreatment of various feedstocks for lactic acid production: detection of sugars, organic acids and furanics in liquid fractions

NARCIS (Netherlands)

Harmsen, P.F.H.; Lips, S.J.J.; Bakker, R.R.C.

2012-01-01

Barley straw, sugarcane bagasse and empty fruit bunches were pretreated under acid- and alkaline conditions. Solid phase was separated from the liquid phase and the concentration of dissolved monomeric sugars, organic acids and furanics was determined. Acid hydrolysis yielded monomeric xylose

Synthesis of DOTA-conjugated multimeric [Tyr3]octreotide peptides via a combination of Cu(I)-catalyzed "click" cycloaddition and thio acid/sulfonyl azide "sulfo-click" amidation and their in vivo evaluation.

NARCIS (Netherlands)

Yim, C.B.; Dijkgraaf, I.; Merkx, R.; Versluis, C.; Eek, A.; Mulder, G.E.; Rijkers, D.T.; Boerman, O.C.; Liskamp, R.M.

2010-01-01

Herein, we describe the design, synthesis, and biological evaluation of a series of DOTA-conjugated monomeric, dimeric, and tetrameric [Tyr(3)]octreotide-based analogues as a tool for tumor imaging and/or radionuclide therapy. These compounds were synthesized using a Cu(I)-catalyzed 1,3-dipolar

Tank Bladders for Advanced Monopropellants, Phase I

Data.gov (United States)

National Aeronautics and Space Administration — In response to RFP S8.04 Spacecraft Propulsion, innovations in propulsion technologies are needed to increase the capabilities of the Science Mission Directorate...

76 FR 7575 - National Cancer Institute; Notice of Meeting

Science.gov (United States)

2011-02-10

... Budget Presentations; Reports of Special Initiatives; RFA and RFP Concept Reviews; and Scientific..., hotel, and airport shuttles will be inspected before being allowed on campus. Visitors will be asked to...

76 FR 62422 - National Cancer Institute; Notice of Meetings

Science.gov (United States)

2011-10-07

... Groups; and Budget Presentations; Reports of Special Initiatives; RFA and RFP Concept Reviews; and... taxicabs, hotel, and airport shuttles will be inspected before being allowed on campus. Visitors will be...

The reversed-field pinch: a compact approach to fusion power

International Nuclear Information System (INIS)

Hagenson, R.L.; Krakowski, R.A.; Bathke, C.G.; Miller, R.L.

1985-01-01

The potential of the reversed-field pinch (RFP) for development into an efficient, compact, copper-coil fusion reactor has been quantified by comprehensive parametric tradeoff studies. This compact system promises to be competitive in size, power density, and cost to alternative energy sources. Conceptual engineering designs that substantiate these promising results have been completed. This 1000 MW(e) (net) design is described along with a detailed rationale and physics/technology assessment for the compact approach to fusion. The RFP presents a robust plasma confinement system capable of providing a range of reactor systems that are compact in both physical size and/or net power output while ensuring acceptable cost and engineering feasibility for a range of assumed physics performance. (author)

Oscillating field current drive for reversed field pinch discharges

International Nuclear Information System (INIS)

Schoenberg, K.F.; Gribble, R.F.; Baker, D.A.

1984-06-01

Oscillating Field Current Drive (OFCD), also known as F-THETA pumping, is a steady-state current-drive technique proposed for the Reversed Field Pinch (RFP). Unlike other current-drive techniques, which employ high-technology, invasive, and power intensive schemes using radio frequency or neutral particle injection, F-THETA pumping entails driving the toroidal and poloidal magnetic field circuits with low-frequency (audio) oscillating voltage sources. Current drive by this technique is a consequence of the strong nonlinear plasma coupling in the RFP. Because of its low frequency and efficient plasma coupling, F-THETA pumping shows excellent promise as a reactor-relevant current-drive technique. A conceptual and computational study of this concept, including its experimental and reactor relevance, is explored in this paper

Theoretical studies of turbulence and anomalous transport in toroidal confinement devices

International Nuclear Information System (INIS)

Terry, P.W.

1990-01-01

The research performed under this grant during the current year has focused on key issues with respect to turbulence and transport in toroidal confinement devices. This work includes theoretical and computational studies of electron thermal confinement which have concentrated on the role of sheared poloidal flow in suppressing turbulence and transport, trapped ion convective cell turbulence and microtearing turbulence; analytical studies of anomalous particle transport and pinch mechanisms, and comparison with experimental measurement; development of the theory of self-consistent radial transport of field-aligned momentum in the tokamak and RFP; and work on other topics (ion temperature gradient driven turbulence, RFP fluctuation theory, coherent structures). Progress and publications in these areas are briefly summarized in this report. 20 refs

Sustainment dynamo reexamined: nonlocal electrical conductivity of plasma in a stochastic magnetic field

International Nuclear Information System (INIS)

Jacobson, A.R.; Moses, R.W.

1984-01-01

The plasma dynamo is both an intriguing and a practical concept. The intrigue derives from attempting to explain naturally occurring and man-made plasmas whose strong field-aligned currents j/sub parallel/ apparently disobey the most naive Ohm's law j/sub parallel/ = sigma/sub parallel/E/sub parallel/. The practical importance derives from the dynamo's role both in formation and in sustainment of reversed-field pinch (RFP) and Spheromak fusion plasmas. We will examine certain features of the documented quasi-steady discharges on ZT-40M, and RFP in apparent need of a sustainment dynamo. We will show that the tail electrons (which carry j/sub parallel/) are probably wandering (along stochastic B Vector-field lines) over much of the minor radius in one mean-free-path

Integrating NRDA and CERCLA environmental evaluations at the Rocky Flats Plant: A case study

International Nuclear Information System (INIS)

Knudsen, T.L.

1993-01-01

The purpose of this paper is to briefly review cleanup regulations in reference to natural resource liability, protection, and restoration; to present a case study on the Rocky Flats Plant (RFP) showing how this DOE facility is approaching the task of integrating the ecological assessment/impact portion of three major regulatory mandates: Resource Conservation and Recovery Act (RCRA), Comprehensive Environmental Response, Compensation and Liability Act (CERCLA), and the National Environmental Policy Act (NEPA) with the Natural Resource Damage Assessment (NRDA) regulations using a flow chart depicting RCRA/CERCLA interim and final actions; to present what has and has not worked at the RFP; and, finally to suggest some technical strategies when planning for remediation and restoration in the NRD process that should be considered

IDAHO BIODIESEL INFRASTRUCTURE PROJECT DOE'S INITIATIVE ON COOPERATIVE PROGRAMS WITH STATES FOR RESEARCH, DEVELOPMENT AND DEMONSTRATION GRANT NO. DE-FC36-02GO12021. Final report

Energy Technology Data Exchange (ETDEWEB)

CROCKETT, JOHN

2006-12-31

The Idaho Energy Division issued a Request for Proposal (RFP) on March 14, 2006, inviting qualified licensed fuel wholesalers, fuel retailers, and vehicle fleet operators to provide proposals to construct and/or install infrastructure for biodiesel utilization in Idaho. The intent was to improve the ability of private and/or non-Federal public entities in Idaho to store, transport, or offer for sale biodiesel within the state. The RFP provided up $100,000 for co-funding the projects with a minimum 50% cash cost match. Four contracts were subsequetnly awarded that resulted in three new bidodiesel storage facilities immediately serving about 45 fueling stations from Sandpoint to Boise. The project also attracted considerable media attention and Idaho became more knowledgeable about biodiesel.

ZTI: Preliminary characterization of an ignition class reversed-field pinch

International Nuclear Information System (INIS)

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

1990-01-01

A preliminary cost-optimized conceptual design of an intermediate-step, ignition-class RFP device (ZTI) for the study of alpha-particle physics in a DT plasma is reported. The ZTI design reflects potentially significant cost savings relative to similar ignition-class tokamaks for device parameters that reside on the path to a viable commercial RFP reactor. Reductions in both device costs and number of steps to commercialization portend a significantly reduced development cost for fusion. The methodology and result and coupling realistic physics, engineering, and cost models through a multi-dimensional optimizer are reported for ZTI, which is a device that would follow the 2--4 MA ZTH on a approx-gt 1996--98 timescale. 15 refs., 7 figs., 2 tabs

The Meteorological Monitoring program at a former nuclear weapons plant

International Nuclear Information System (INIS)

Maxwell, D.R.; Bowen, B.M.

1994-01-01

The purpose of the Meteorological Monitoring program at Rocky Flats Plant (RFP) is to provide meteorological information for use in assessing the transport, and diffusion, and deposition of effluent actually or potentially released into the atmosphere by plant operations. Achievement of this objective aids in protecting health and safety of the public, employees, and environment, and directly supports Emergency Response programs at RFP. Meteorological information supports the design of environmental monitoring networks for impact assessments, environmental surveillance activities, remediation activities, and emergency responses. As the mission of the plant changes from production of nuclear weapons parts to environmental cleanup and economic development, smaller releases resulting from remediation activities become more likely. These possible releases could result from airborne fugitive dust, evaporation from collection ponds, or grass fires

Economically attractive features of steady-state neoclassical reversed field pinch equilibrium with low aspect ratio

International Nuclear Information System (INIS)

Shiina, S.; Yagi, Y.; Sugimoto, H.; Ashida, H.; Hirano, Y.; Koguchi, H.; Sakakita, H.; Taguchi, M.; Nagamine, Y.; Osanai, Y.; Saito, K.; Watanabe, M.; Aizawa, M.

2005-01-01

Dominant plasma self-induced current equilibrium is achieved together with the high β for the steady-state neoclassical reversed field pinch (RFP) equilibrium with low aspect ratio by broadening the plasma pressure profile. The RF-driven current, when the safety factor is smaller than unity, is much less than the self-induced current, which dominates (96%) the toroidal current. This neoclassical RFP equilibrium has strong magnetic shear or a high-stability beta (β t = 63%) due to its hollow current profile. It is shown that the obtained equilibrium is close to the relaxed-equilibrium state with a minimum energy, and is also robust against microinstabilities. These attractive features allow the economical design of compact steady-state fusion power plants with low cost of electricity (COE). (author)

A comprehensive theory of the equilibria in a tokamak and a reversed field pinch

International Nuclear Information System (INIS)

Chiyoda, Katsuji

1996-01-01

The equilibrium configuration of a tokamak is analysed by the equilibrium equations derived for analysing a reversed field pinch (RFP). The expressions of the magnetic field and the toroidal shift in the internal plasma region and the external vacuum region are obtained. The expressions in the vacuum region become the Shafranov's expressions, when the plasma-center coordinates is used. Discontinuities of the equilibrium quantities are considered. It is concluded that the equilibrium equations are applicable also to the tokamak plasma and that the difference of the equilibria between the tokamak and the RFP stems from the choices of the pressure and the toroidal current function. A feature of our theory is that any ordering to the safety factor is not imposed. (author)

Influence of osmolarity of contrast medium and saline flush on computed tomography angiography: Comparison of monomeric and dimeric iodinated contrast media with different iodine concentrations at an identical iodine delivery rate

International Nuclear Information System (INIS)

Kishimoto, Miori; Doi, Shoko; Shimizu, Junichiro; Lee, Ki-Ja; Iwasaki, Toshiroh; Miyake, Yoh-Ichi; Yamada, Kazutaka

2010-01-01

Purpose: To evaluate the influence of osmolarity of iodinated contrast media and saline flush on the contrast effect in thoracic computed tomography angiography (CTA) at an identical iodine delivery rate (IDR). Materials and methods: Seven beagles were used in a cross-over experiment. The contrast media used were iohexol 350 mgI/ml (IOH350; osmolarity 844 mmol/kg) and iodixanol 320 mgI/ml (IDX320; osmolarity 290 mmol/kg). Each contrast medium was administered to groups with and without saline flush at 40.0 mgI/kg/s for all experiments. Dynamic CT scanning was performed at the ninth thoracic vertebra level. The peak value, area under the curve (AUC), and time to peak (TTP) were calculated from the time attenuation curves of the pulmonary artery and aorta. Results: There was no significant difference between IOH350 and IDX320 with or without saline flush in the peak values for the pulmonary artery and aorta. AUC was significantly higher in groups with saline flush for both contrast media and arteries (p < 0.05) with no significant difference between contrast media. TTP was significantly longer in groups with saline flush than without saline flush for both contrast media and arteries (p < 0.05), with no significant difference between contrast media. Conclusions: There were no significant differences in the contrast effects of monomeric IOH350 and dimeric IDX320 in thoracic CTA when used at an identical IDR. Moreover, saline flush prolonged the peak duration at 600 mgI/kg.

Chemistry of organogold compounds I. Syntheses and properties of dihalogold(III) N,N-dialkyldithiocarbamates and dialkylgold(III) N,N-dialkyldithiocarbamates

NARCIS (Netherlands)

Blaauw, H.J.A.; Nivard, R.J.F.; Kerk, G.J.M. van der

Monomeric dialkylgold(III) N,N-dialkyldithiocarbamates (R2AuSSCNR'2) can be prepared in good yields from monomeric dibromogold(III) N,N-dialkyldithiocarbamates (Br2AuSSCNR'2) and alkylmagnesium halides or dialkylcadmium compounds. In addition N-ethylpiperidinium tetrachloroaurate can be converted

75 FR 63494 - National Cancer Institute; Notice of Meeting

Science.gov (United States)

2010-10-15

... Group(s); and Budget Presentations; Reports of Special Initiatives; RFA and RFP Concept Reviews; and... taxicabs, hotel, and airport shuttles will be inspected before being allowed on campus. Visitors will be...

78 FR 5192 - National Cancer Institute; Notice of Meeting

Science.gov (United States)

2013-01-24

... Review Group(s); and Budget Presentations; Reports of Special Initiatives; RFA and RFP Concept Reviews... entrance onto the NIH campus. All visitor vehicles, including taxicabs, hotel, and airport shuttles will be...

Structural Variation of Alpha-synuclein with Temperature by a Coarse-grained Approach with Knowledge-based Interactions (Postprint)

Science.gov (United States)

2015-07-01

Alzheimer disease. ASN1,3 has been extensively studied as an intrinsically disordered (unstructured) protein with a monomeric conformation compa- rable to a...changes in these physical quantities occur by avoiding the low and high temperature extremes. Typical snapshots from the simulations as a function of...mobile as the residue-residue interactions become less impor- tant and the protein assumes a self- avoiding walk (SAW) or random coil conformation (vida

Two-dimensional condensation of pyrimidine oligonucleotides during their self-assemblies at mercury based surfaces

Energy Technology Data Exchange (ETDEWEB)

Hason, Stanislav [Institute of Biophysics, Academy of Sciences of the Czech Republic, v.v.i. Kralovopolska 135, CZ-612 65 Brno (Czech Republic)], E-mail: hasons@ibp.cz; Vetterl, Vladimir [Institute of Biophysics, Academy of Sciences of the Czech Republic, v.v.i. Kralovopolska 135, CZ-612 65 Brno (Czech Republic); Fojta, Miroslav [Institute of Biophysics, Academy of Sciences of the Czech Republic, v.v.i. Kralovopolska 135, CZ-612 65 Brno (Czech Republic)], E-mail: fojta@ibp.cz

2008-02-15

For the first time it is shown that homopyrimidine oligodeoxynucleotides (ODNs) adsorbed at mercury or amalgam electrode surface can condensate upon applying negative potentials (around -1.35 V vs. Ag/AgCl/3M KCl). This 2D condensation resulted in formation of capacitance pits on the C-E curves resembling those observed earlier with monomeric nucleic acid bases, nucleosides and nucleotides. Differences in behavior of the condensed layers of dT{sub 30} and dC{sub 30} ODNs, reflecting different physico-chemical and electrochemical properties of thymine and cytosine, were observed. Formation of the ODN condensed film involved reorientation of the oligonucleotide molecules firmly adsorbed at the electrode and took place even in the absence of any ODN in the bulk of solution. Homopurine ODNs did not form these two-dimensional (2D) condensed monolayers under the same conditions. A preliminary thermodynamic analysis of the condensed ODN layers is presented.

Origins of fluorescence in evolved bacteriophytochromes.

Science.gov (United States)

Bhattacharya, Shyamosree; Auldridge, Michele E; Lehtivuori, Heli; Ihalainen, Janne A; Forest, Katrina T

2014-11-14

Use of fluorescent proteins to study in vivo processes in mammals requires near-infrared (NIR) biomarkers that exploit the ability of light in this range to penetrate tissue. Bacteriophytochromes (BphPs) are photoreceptors that couple absorbance of NIR light to photoisomerization, protein conformational changes, and signal transduction. BphPs have been engineered to form NIR fluorophores, including IFP1.4, Wi-Phy, and the iRFP series, initially by replacement of Asp-207 by His. This position was suggestive because its main chain carbonyl is within hydrogen-bonding distance to pyrrole ring nitrogens of the biliverdin chromophore, thus potentially functioning as a crucial transient proton sink during photoconversion. To explain the origin of fluorescence in these phytofluors, we solved the crystal structures of IFP1.4 and a comparison non-fluorescent monomeric phytochrome DrCBDmon. Met-186 and Val-288 in IFP1.4 are responsible for the formation of a tightly packed hydrophobic hub around the biliverdin D ring. Met-186 is also largely responsible for the blue-shifted IFP1.4 excitation maximum relative to the parent BphP. The structure of IFP1.4 revealed decreased structural heterogeneity and a contraction of two surface regions as direct consequences of side chain substitutions. Unexpectedly, IFP1.4 with Asp-207 reinstalled (IFPrev) has a higher fluorescence quantum yield (∼9%) than most NIR phytofluors published to date. In agreement, fluorescence lifetime measurements confirm the exceptionally long excited state lifetimes, up to 815 ps, in IFP1.4 and IFPrev. Our research helps delineate the origin of fluorescence in engineered BphPs and will facilitate the wide-spread adoption of phytofluors as biomarkers. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

A redox-dependent dimerization switch regulates activity and tolerance for reactive oxygen species of barley seed glutathione peroxidase

DEFF Research Database (Denmark)

Navrot, Nicolas; Skjoldager, Nicklas; Bunkenborg, Jakob

2015-01-01

Monomeric and dimeric forms of recombinant barley (Hordeum vulgare subsp. vulgare) glutathione peroxidase 2 (HvGpx2) are demonstrated to display distinctly different functional properties in vitro. Monomeric HvGpx2 thus has five fold higher catalytic efficiency than the dimer towards tert-butyl h...

Shark immunity bites back: affinity maturation and memory response in the nurse shark, Ginglymostoma cirratum.

Science.gov (United States)

Dooley, Helen; Flajnik, Martin F

2005-03-01

The cartilaginous fish are the oldest phylogenetic group in which all of the molecular components of the adaptive immune system have been found. Although early studies clearly showed that sharks could produce an IgM-based response following immunization, evidence for memory, affinity maturation and roles for the other isotypes (notably IgNAR) in this group remained inconclusive. The data presented here illustrate that the nurse shark (Ginglymostoma cirratum) is able to produce not only an IgM response, but we also show for the first time a highly antigen-specific IgNAR response. Additionally, under appropriate conditions, a memory response for both isotypes can be elicited. Analysis of the response shows differential expression of pentameric and monomeric IgM. Pentameric IgM provides the 'first line of defense' through high-avidity, low-affinity interaction with antigen. In contrast, monomeric IgM and IgNAR seem responsible for the specific, antigen-driven response. We propose the presence of distinct lineages of B cells in sharks. As there is no conventional isotype switching, each lineage seems pre-determined to express a single isotype (IgM versus IgNAR). However, our data suggest that there may also be specific lineages for the different forms (pentameric versus monomeric) of the IgM isotype.

Advanced Design Composite Aircraft

Science.gov (United States)

1976-02-01

materials. Preliminary vehicle syn- thesis was based on the RFP mission and the initial ADCA configuration design cycle was started. Aerodynamic, mass...i. ■.■.»■u.u.ill. ...LL., , ,,„I,II,„IUUBI u i i. Mini » .11 wrmmmmmmmmm- ill m<* BUCKLING COEFFICIENT FOR SIMPLY SUPPORTED FLAT SANDWICH PANELS...SecmoKj Tl PlC ^L SS-CTIOSJ A-A fouu Sire CvRAPHnE/^KilfJ PtU-feR f.»»..-—..I....... - ■■■ ■*■- • -- ^- TZlll’, ’-TTT

Succinic Acid as a Byproduct in a Corn-based Ethanol Biorefinery

Energy Technology Data Exchange (ETDEWEB)

MBI International

2007-12-31

MBI endeavored to develop a process for succinic acid production suitable for integration into a corn-based ethanol biorefinery. The project investigated the fermentative production of succinic acid using byproducts of corn mill operations. The fermentation process was attuned to include raw starch, endosperm, as the sugar source. A clean-not-sterile process was established to treat the endosperm and release the monomeric sugars. We developed the fermentation process to utilize a byproduct of corn ethanol fermentations, thin stillage, as the source of complex nitrogen and vitamin components needed to support succinic acid production in A. succinogenes. Further supplementations were eliminated without lowering titers and yields and a productivity above 0.6 g l-1 hr-1was achieved. Strain development was accomplished through generation of a recombinant strain that increased yields of succinic acid production. Isolation of additional strains with improved features was also pursued and frozen stocks were prepared from enriched, characterized cultures. Two recovery processes were evaluated at pilot scale and data obtained was incorporated into our economic analyses.

New transmission planning methodology for requesting proposals for wind generation

Science.gov (United States)

Isaacs, Andrew L.

The increasing interest in renewable energy technologies during the last decade has caused conventional transmission and generation expansion planning methodologies to be strained and in some cases abandoned. This is due both to the quantity of generator interconnection requests and the constraints imposed by deregulated energy industry structures. One technique used to control the influx of renewable generation while maintaining competitive principles is a Request for Proposals (RFP). However, lack of transmission planning due to a disconnection between generation and transmission owners, difficulty in identifying viable projects, and high risk for proponents stand as obstacles to the goals of an RFP. This research proposes a procedure which minimizes the effect of these obstacles; meeting the purchaser requirements for low price and combining conventional planning concepts with feedback from competitive structures. The general features of the method include definition of generation limits and study area, expansion plan design, transmission cost evaluation, optimal selection of requested generation levels, and final selection of successful proponents. The method is shown to be effective in creating an RFP where proponents are well-informed and provided with cost certainty to reduce bid price, buyers are able to determine end costs of their energy, and good expansion planning principles are maintained. A case study using a real system in New Mexico demonstrates these concepts.

Renewable Energy Contract Development Best Practices

Science.gov (United States)

This page provides information on how available resources can help organizations effectively navigate the solar contract development process with the goal of RFP and contract transparency, completeness, and accountability of all involved parties.

Microalgae Compositional Analysis Laboratory Procedures | Bioenergy | NREL

Science.gov (United States)

hydrolyze the polymeric forms of carbohydrates in algal biomass into monomeric subunits. The monomers are polymeric structural and storage carbohydrates into monomeric sugars. A range of sequential and optimized present in many forms and play various roles within an algal cell, from cell membrane phospholipids to

Photophysics and photochemistry of photoreactivation

International Nuclear Information System (INIS)

Sutherland, J.C.

1977-01-01

Photoreactivating enzyme (PRE) monomerizes cyclobutyl pyrimidine dimers formed in DNA by UV light (lambda < 300 nm). The enzyme requires near UV and visible wavelengths (300 < lambda < 600 nm) for activity. Possible mechanisms of action of the PRE are suggested by non-enzymatic processes in which pyrimidine dimers are monomerized by UV and visible light. Two such non-enzymatic processes are (a) photolysis of dimers resulting from direct absorption of UV, and (b) sensitized monomerization involving charge transfer complexes. Several lines of evidence suggest that the mechanism of action of the PRE more closely resembles (b) than (a). Recent experiments on the PRE from E.coli revealed the presence of new long wavelength absorption which may indicate the presence of a ground state complex. The known ability of PRE to monomerize dimers of thymine, cytosine and uracil suggests that the carbonyl groups at 2 position of the pyrimidine ring may be important in the interaction between enzyme and dimer. (author)

Photophysics and photochemistry of photoreactivation

Energy Technology Data Exchange (ETDEWEB)

Sutherland, J C [California Univ., Irvine (USA)

1977-05-01

Photoreactivating enzyme (PRE) monomerizes cyclobutyl pyrimidine dimers formed in DNA by uv light (lambda < 300 nm). The enzyme requires near UV and visible wavelengths (300 < lambda < 600 nm) for activity. Possible mechanisms of action of the PRE are suggested by non-enzymatic processes in which pyrimidine dimers are monomerized by UV and visible light. Two such non-enzymatic processes are (a) photolysis of dimers resulting from direct absorption of UV, and (b) sensitized monomerization involving charge transfer complexes. Several lines of evidence suggest that the mechanism of action of the PRE more closely resembles (b) than (a). Recent experiments on the PRE from E.coli revealed the presence of new long wavelength absorption which may indicate the presence of a ground state complex. The known ability of PRE to monomerize dimers of thymine, cytosine and uracil suggests that the carbonyl groups at 2 position of the pyrimidine ring may be important in the interaction between enzyme and dimer.

Why can a gold salt react as a base?

Science.gov (United States)

Anania, Mariarosa; Jašíková, Lucie; Jašík, Juraj; Roithová, Jana

2017-09-26

This study shows that gold salts [(L)AuX] (L = PMe 3 , PPh 3 , JohnPhos, IPr; X = SbF 6 , PF 6 , BF 4 , TfO, Tf 2 N) act as bases in aqueous solutions and can transform acetone to digold acetonyl complexes [(L) 2 Au 2 (CH 2 COCH 3 )] + without any additional base present in solution. The key step is the formation of digold hydroxide complexes [(L) 2 Au 2 (OH)] + . The kinetics of the formation of the digold complexes and their mutual transformation is studied by electrospray ionization mass spectrometry and the delayed reactant labelling method. We show that the formation of digold hydroxide is the essential first step towards the formation of the digold acetonyl complex, the reaction is favoured by more polar solvents, and the effect of counter ions is negligible. DFT calculations suggest that digold hydroxide and digold acetonyl complexes can exist in solution only due to the stabilization by the interaction with two gold atoms. The reaction between the digold hydroxide and acetone proceeds towards the dimer {[(L)Au(OH)]·[(L)Au(CH 3 COCH 3 )] + }. The monomeric units interact at the gold atoms in the perpendicular arrangement typical of the gold clusters bound by the aurophilic interaction. The hydrogen is transferred within the dimer and the reaction continues towards the digold acetonyl complex and water.

Fabrication and bonding of thiol-ene-based microfluidic devices

International Nuclear Information System (INIS)

Sikanen, Tiina M; Moilanen, Maria-Elisa; Lafleur, Josiane P; Zhuang, Guisheng; Jensen, Thomas G; Kutter, Jörg P

2013-01-01

In this work, the bonding strength of microchips fabricated by thiol-ene free-radical polymerization was characterized in detail by varying the monomeric thiol/allyl composition from the stoichiometric ratio (1:1) up to 100% excess of thiol (2:1) or allyl (1:2) functional groups. Four different thiol-ene to thiol-ene bonding combinations were tested by bonding: (i) two stoichiometric layers, (ii) two layers bearing complementary excess of thiols and allyls, (iii) two layers both bearing excess of thiols, or (iv) two layers both bearing excess of allyls. The results showed that the stiffness of the cross-linked polymer plays the most crucial role regarding the bonding strength. The most rigid polymer layers were obtained by using the stoichiometric composition or an excess of allyls, and thus, the bonding combinations (i) and (iv) withstood the highest pressures (up to the cut-off value of 6 bar). On the other hand, excess of thiol monomers yielded more elastic polymer layers and thus decreased the pressure tolerance for bonding combinations (ii) and (iii). By using monomers with more thiol groups (e.g. tetrathiol versus trithiol), a higher cross-linking ratio, and thus, greater stiffness was obtained. Surface characterization by infrared spectroscopy confirmed that the changes in the monomeric thiol/allyl composition were also reflected in the surface chemistry. The flexibility of being able to bond different types of thiol-enes together allows for tuning of the surface chemistry to yield the desired properties for each application. Here, a capillary electrophoresis separation is performed to demonstrate the attractive properties of stoichiometric thiol-ene microchips. (technical note)