WorldWideScience

Sample records for radiochemically modified protein

  1. Radiochemicals

    International Nuclear Information System (INIS)

    1980-01-01

    In this catalogue those radioactive chemicals for research are listed which are produced by the Radiochemical Centre Amersham and our laboratories at Brunswick. The dates given for each product can understandably only be limited within the framework of such a catalogue. Additional dates and references to application technique can be obtained from us any time. Our programme is continually updated by new products. If a compound not listed in the catalogue should be required we ask for inquiry. Our working team for special syntheses will try to produce it according to our possibilities and our requirements. (orig.) [de

  2. Protein binding studies with radiolabeled compounds containing radiochemical impurities. Equilibrium dialysis versus dialysis rate determination

    DEFF Research Database (Denmark)

    Honoré, B

    1987-01-01

    The influence of radiochemical impurities in dialysis experiments with high-affinity ligands is investigated. Albumin binding of labeled decanoate (97% pure) is studied by two dialysis techniques. It is shown that equilibrium dialysis is very sensitive to the presence of impurities resulting...

  3. Radiochemical procedures

    International Nuclear Information System (INIS)

    Lyon, W.S.

    1982-01-01

    The modern counting instrumentation has largely obviated the need for separation processes in the radiochemical analysis but problems in low-level radioactivity measurement, environmental-type analyses, and special situations caused in the last years a renaissance of the need for separation techniques. Most of the radiochemical procedures, based on the classic works of the Manhattan Project chemists of the 1940's, were published in the National Nuclear Energy Series (NNES). Improvements such as new solvent extraction and ion exchange separations have been added to these methods throughout the years. Recently the Los Alamos Group have reissued their collected Radiochemical Procedures containing a short summary and review of basic inorganic chemistry - 'Chemistry of the Elements on the Basis of Electronic Configuration'. (A.L.)

  4. Adhesives from modified soy protein

    Science.gov (United States)

    Sun, Susan [Manhattan, KS; Wang, Donghai [Manhattan, KS; Zhong, Zhikai [Manhattan, KS; Yang, Guang [Shanghai, CN

    2008-08-26

    The present invention provides useful adhesive compositions having similar adhesive properties to conventional UF and PPF resins. The compositions generally include a protein portion and modifying ingredient portion selected from the group consisting of carboxyl-containing compounds, aldehyde-containing compounds, epoxy group-containing compounds, and mixtures thereof. The composition is preferably prepared at a pH level at or near the isoelectric point of the protein. In other preferred forms, the adhesive composition includes a protein portion and a carboxyl-containing group portion.

  5. Radiochemical methods

    International Nuclear Information System (INIS)

    Geary, W.J.

    1986-01-01

    This little volume is one of an extended series of basic textbooks on analytical chemistry produced by the Analytical Chemistry by Open Learning project in the UK. Prefatory sections explain its mission, and how to use the Open Learning format. Seventeen specific sections organized into five chaptrs begin with a general discussion of nuclear properties, types, and laws of nuclear decay and proceeds to specific discussions of three published papers (reproduced in their entirety) giving examples of radiochemical methods which were discussed in the previous chapter. Each section begins with an overview, contains one or more practical problems (called self-assessment questions or SAQ's), and concludes with a summary and a list of objectives for the student. Following the main body are answers to the SAQ's, and several tables of physical constants, SI prefixes, etc. A periodic table graces the inside back cover

  6. Evaluation of radiochemical data usability

    International Nuclear Information System (INIS)

    Paar, J.G.; Porterfield, D.R.

    1997-04-01

    This procedure provides a framework for implementation of radiochemical data verification and validation for environmental remediation activities. It has been developed through participation of many individuals currently involved in analytical radiochemistry, radiochemical validation, and validation program development throughout the DOE complex. It should be regarded as a guidance to use in developing an implementable radiochemical validation strategy. This procedure provides specifications for developing and implementing a radiochemical validation methodology flexible enough to allow evaluation of data useability for project-specific Data Quality Objectives (DQO). Data produced by analytical methods for which this procedure provides limited guidance are classified as open-quotes non-routineclose quotes radionuclides and methods, and analyses by these methods may necessitate adoption of modified criteria from this procedure

  7. Radiochemicals in biomedical research

    International Nuclear Information System (INIS)

    Evans, E.A.; Oldham, K.G.

    1988-01-01

    This volume describes the role of radiochemicals in biomedical research, as tracers in the development of new drugs, their interaction and function with receptor proteins, with the kinetics of binding of hormone - receptor interactions, and their use in cancer research and clinical oncology. The book also aims to identify future trends in this research, the main objective of which is to provide information leading to improvements in the quality of life, and to give readers a basic understanding of the development of new drugs, how they function in relation to receptor proteins and lead to a better understanding of the diagnosis and treatment of cancers. (author)

  8. Modified Protein Improves Vitiligo Symptoms in Mice

    Science.gov (United States)

    ... Vitiligo Symptoms in Mice Spotlight on Research Modified Protein Improves Vitiligo Symptoms in Mice By Colleen Labbe, ... D., Ph.D., Rush University. Altering a key protein involved in the development of vitiligo may protect ...

  9. Chromatin-modifying proteins in cancer

    DEFF Research Database (Denmark)

    Fog, Cathrine K; Jensen, Klaus T; Lund, Anders Henrik

    2007-01-01

    -despite the fact that all cells in the organism contain the same genetic information. A large amount of data gathered over the last decades has demonstrated that deregulation of chromatin-modifying proteins is etiologically involved in the development and progression of cancer. Here we discuss how epigenetic...... alterations influence cancer development and review known cancer-associated alterations in chromatin-modifying proteins....

  10. How proteins modify water dynamics

    Science.gov (United States)

    Persson, Filip; Söderhjelm, Pär; Halle, Bertil

    2018-06-01

    Much of biology happens at the protein-water interface, so all dynamical processes in this region are of fundamental importance. Local structural fluctuations in the hydration layer can be probed by 17O magnetic relaxation dispersion (MRD), which, at high frequencies, measures the integral of a biaxial rotational time correlation function (TCF)—the integral rotational correlation time. Numerous 17O MRD studies have demonstrated that this correlation time, when averaged over the first hydration shell, is longer than in bulk water by a factor 3-5. This rotational perturbation factor (RPF) has been corroborated by molecular dynamics simulations, which can also reveal the underlying molecular mechanisms. Here, we address several outstanding problems in this area by analyzing an extensive set of molecular dynamics data, including four globular proteins and three water models. The vexed issue of polarity versus topography as the primary determinant of hydration water dynamics is resolved by establishing a protein-invariant exponential dependence of the RPF on a simple confinement index. We conclude that the previously observed correlation of the RPF with surface polarity is a secondary effect of the correlation between polarity and confinement. Water rotation interpolates between a perturbed but bulk-like collective mechanism at low confinement and an exchange-mediated orientational randomization (EMOR) mechanism at high confinement. The EMOR process, which accounts for about half of the RPF, was not recognized in previous simulation studies, where only the early part of the TCF was examined. Based on the analysis of the experimentally relevant TCF over its full time course, we compare simulated and measured RPFs, finding a 30% discrepancy attributable to force field imperfections. We also compute the full 17O MRD profile, including the low-frequency dispersion produced by buried water molecules. Computing a local RPF for each hydration shell, we find that the

  11. Radiochemical Processing Laboratory (RPL)

    Data.gov (United States)

    Federal Laboratory Consortium — The Radiochemical Processing Laboratory (RPL)�is a scientific facility funded by DOE to create and implement innovative processes for environmental clean-up and...

  12. Radiochemical procedures and techniques

    International Nuclear Information System (INIS)

    Flynn, K.

    1975-04-01

    A summary is presented of the radiochemical procedures and techniques currently in use by the Chemistry Division Nuclear Chemistry Group at Argonne National Laboratory for the analysis of radioactive samples. (U.S.)

  13. Radiochemical solar neutrino experiments

    International Nuclear Information System (INIS)

    Rich, R.; Spiro, M.

    1993-01-01

    This review covers the three presently running radiochemical solar neutrino experiments, namely the Chlorine, SAGE, and GALLEX experiments. The focus of the review is on a discussion of statistical consistency checks of the available data. The chlorine radiochemical experiment is conceptually simple and shows no strong indication of any statistical anomalies. It still forms the basis of the solar neutrino problem. Each of the two gallium experiments show internal statistical consistency. SAGE's recent preliminary results are consistent with the published GALLEX results. If this convergence is confirmed by a more definitive analysis, this would suggest that the combined result of the two gallium experiments, SAGE and GALLEX, be used for comparisons with theoretical expectations. 5 refs., 15 figs

  14. The Protein-Sparing Modified Fast Diet

    Directory of Open Access Journals (Sweden)

    Marwan Bakhach MD

    2016-01-01

    Full Text Available Objectives: The protein-sparing modified fast (PSMF is a rigorous way of rapidly losing a large amount of weight. Although adult studies have shown the PSMF to be effective, data in adolescents are lacking. The aim of this study was to determine the efficacy and safety of the PSMF in severely obese adolescents. Methods: 12 subjects who were evaluated in the Obesity Management Program at the Cleveland Clinic from 2011 to 2014 were included. The subjects were initiated on the PSMF after failing other conventional methods of weight loss. Once the goal weight was achieved, subjects were transitioned to the refeeding phase for weight maintenance. Results: Follow-up was scheduled at 3-month (11 patients and 6-month (6 patients intervals. At the 6-month follow-up visit, the average weight loss was 11.19 kg (95% confidence interval = -5.4, -27.8, P = .028, with average of 9.8% from baseline. Fifty percent of subjects had >5% weight loss and 20% had >10% weight loss. Four patients were lost to the follow-up (40%. An improvement was noted in total cholesterol and high-density lipoprotein. Due to a small sample size these results were not statistically significant. Side effects reported by subjects were mild dehydration due to nausea (2 patients, decreased energy (1 patient, and transient labile mood (1 patient. No life-threatening side effects were reported. Conclusion: Our results show that the PSMF diet can be used as an effective and safe method in the outpatient setting for rapid weight loss in adolescents with severe obesity.

  15. Photochemistry of modified proteins benzophenone-containing bovine serum albumin

    International Nuclear Information System (INIS)

    Mariano, P.S.; Glover, G.I.; Wilkinson, T.J.

    1976-01-01

    The results of exploratory and mechanistic studies of the photochemistry of poly-p-benzoyl-acetimido-bovine serum albumin, a modified protein containing photoreactive and photosensitizing groups, are reported. Specifically described are recent findings concerning (1) the synthesis and characterization of a modified bovine serum albumin that contains benzophenone-like moieties, (2) the photochemistry of this modified protein which appeared to involve photoreductive coupling of the benzophenone chromophores to the protein backbone, and (3) triplet energy transfer from modified bovine serum albumin to small molecule acceptors resulting in quenching of the photoreaction. (author)

  16. Proteins mediating intra- and intercellular transport of lipids and lipid-modified proteins

    NARCIS (Netherlands)

    Neumann, S.

    2008-01-01

    Proteins mediating intra- and intercellular transport of lipids and lipid-modified proteins In this thesis, I studied the intra- and intercellular transport of lipidic molecules, in particular glycosphingolipids and lipid-modified proteins. The first part focuses on the intracellular transport of

  17. 11th radiochemical conference

    International Nuclear Information System (INIS)

    Prasil, Z.

    1987-01-01

    The conference met in four sesions which discussed: Separation methods, Radioanalytical methods, Labelled compounds and Miscellaneous. The first session discussed extraction methods, ion exchange and chromatographic separation of radioisotopes. The second session heard papers on the application of these methods, e.g., in geochemistry, on the use of radioactive tracers in radiochemical analysis and the use of activation analysis in the determination of trace elements. The third session heard papers on the preparation of labelled organic compounds with isotopes 3 H, 14 C, radioiodine and 32 P, on the preparation of RIA kits and on the chemistry and radiopharmacology of technetium compounds. The other contributions which could not be heard in any of the three sessions discussed, e.g., the preparation of elements on the cyclotron and microtron, the production of a new 99m Tc-generator, the participation of the IAEA in processing low- and medium-level radioactive wastes, etc. (E.S.)

  18. Radiochemical synthesis of etomoxir

    Energy Technology Data Exchange (ETDEWEB)

    Abbas, Hafiz G. [Institute of Nuclear Medicine and Oncology (INMOL), New Campus Road, Lahore (Pakistan); Yunus, M. [University of the Punjab, New Campus Road, Lahore (Pakistan); Feinendegen, Ludwig E., E-mail: feinendegen@gmx.ne [Department of Nuclear Medicine, Heinrich-Heine University Duesseldorf, Wannental 45, 88131 Lindau (Germany)

    2011-02-15

    Sodium 2-{l_brace}6-(4-chlorophenoxy)hexyl{r_brace}oxirane-2-carboxylate (Etomoxir) inhibits transport of fatty acids via the carnitine shuttle into mitochondria of muscle cells and prevents long chain fatty acids from providing energy through {beta}-oxidation especially for muscle contraction. The objective of this synthesis is to develop a method for radioiodination of Etomoxir in order to explore its potential in diagnostic metabolic studies and molecular imaging. Thus, a method is described for the radiochemical synthesis and purification of ethyl 2-{l_brace}6-(4-[{sup 131}I]iodophenoxy)hexyl{r_brace}oxirane-2-carboxylate (3) and 2-{l_brace}6-(4-[{sup 131}I]iodo-phenoxy)hexyl{r_brace}oxirane-2-carboxylic acid (4). For the synthesis of these new agents, ethyl 2-{l_brace}6-(4-bromophenoxy)hexyl{r_brace}oxirane-2-carboxylate (1) and 2-{l_brace}6-(4-bromophenoxy)hexyl{r_brace}oxirane-2-carboxylic acid (2) were refluxed with [{sup 131}I]NaI in the presence of anhydrous acetone at a temperature of 80 {sup o}C and 90 {sup o}C for a period of 3-4 hours, respectively. The method of radiolabeling, based on the nucleophilic exchange reaction, resulted in a radiochemical yield of 43% and 67% for compounds 3 and 4, respectively. This paper reports on the labeling of etomoxir with radioiodine as {sup 124}I labeled etomoxir may be of great importance in molecular imaging.

  19. Huntingtin interacting proteins are genetic modifiers of neurodegeneration.

    Directory of Open Access Journals (Sweden)

    Linda S Kaltenbach

    2007-05-01

    Full Text Available Huntington's disease (HD is a fatal neurodegenerative condition caused by expansion of the polyglutamine tract in the huntingtin (Htt protein. Neuronal toxicity in HD is thought to be, at least in part, a consequence of protein interactions involving mutant Htt. We therefore hypothesized that genetic modifiers of HD neurodegeneration should be enriched among Htt protein interactors. To test this idea, we identified a comprehensive set of Htt interactors using two complementary approaches: high-throughput yeast two-hybrid screening and affinity pull down followed by mass spectrometry. This effort led to the identification of 234 high-confidence Htt-associated proteins, 104 of which were found with the yeast method and 130 with the pull downs. We then tested an arbitrary set of 60 genes encoding interacting proteins for their ability to behave as genetic modifiers of neurodegeneration in a Drosophila model of HD. This high-content validation assay showed that 27 of 60 orthologs tested were high-confidence genetic modifiers, as modification was observed with more than one allele. The 45% hit rate for genetic modifiers seen among the interactors is an order of magnitude higher than the 1%-4% typically observed in unbiased genetic screens. Genetic modifiers were similarly represented among proteins discovered using yeast two-hybrid and pull-down/mass spectrometry methods, supporting the notion that these complementary technologies are equally useful in identifying biologically relevant proteins. Interacting proteins confirmed as modifiers of the neurodegeneration phenotype represent a diverse array of biological functions, including synaptic transmission, cytoskeletal organization, signal transduction, and transcription. Among the modifiers were 17 loss-of-function suppressors of neurodegeneration, which can be considered potential targets for therapeutic intervention. Finally, we show that seven interacting proteins from among 11 tested were able to

  20. Radiochemical methods to enhance efficiency of α-spectral measurements

    International Nuclear Information System (INIS)

    Silkina, G.P.; Artem'ev, O.I.

    2001-01-01

    The paper describes possible ways to improve a plutonium radiochemical separation technique developed in the Khlopin Radium Institute and modify it to account for the site-specific features of samples from the former Semipalatinsk test site (STS) and enhance the alpha spectrometry efficiency.The paper describes possible ways to improve a plutonium radiochemical separation technique developed in the Khlopin Radium Institute and modify it to account for the site-specific features of samples from the former Semipalatinsk test site (STS) and enhance the alpha spectrometry efficiency. (author)

  1. A modified Poisson-Boltzmann equation applied to protein adsorption.

    Science.gov (United States)

    Gama, Marlon de Souza; Santos, Mirella Simões; Lima, Eduardo Rocha de Almeida; Tavares, Frederico Wanderley; Barreto, Amaro Gomes Barreto

    2018-01-05

    Ion-exchange chromatography has been widely used as a standard process in purification and analysis of protein, based on the electrostatic interaction between the protein and the stationary phase. Through the years, several approaches are used to improve the thermodynamic description of colloidal particle-surface interaction systems, however there are still a lot of gaps specifically when describing the behavior of protein adsorption. Here, we present an improved methodology for predicting the adsorption equilibrium constant by solving the modified Poisson-Boltzmann (PB) equation in bispherical coordinates. By including dispersion interactions between ions and protein, and between ions and surface, the modified PB equation used can describe the Hofmeister effects. We solve the modified Poisson-Boltzmann equation to calculate the protein-surface potential of mean force, treated as spherical colloid-plate system, as a function of process variables. From the potential of mean force, the Henry constants of adsorption, for different proteins and surfaces, are calculated as a function of pH, salt concentration, salt type, and temperature. The obtained Henry constants are compared with experimental data for several isotherms showing excellent agreement. We have also performed a sensitivity analysis to verify the behavior of different kind of salts and the Hofmeister effects. Copyright © 2017 Elsevier B.V. All rights reserved.

  2. 4-Hydroxyhexenal- and 4-Hydroxynonenal-Modified Proteins in Pterygia

    Directory of Open Access Journals (Sweden)

    Ichiya Sano

    2013-01-01

    Full Text Available Oxidative stress has been suspected of contributing to the pathogenesis of pterygia. We evaluated the immunohistochemical localization of the markers of oxidative stress, that is, the proteins modified by 4-hydroxyhexenal (4-HHE and 4-hydroxynonenal (4-HNE, which are reactive aldehydes derived from nonenzymatic oxidation of n-3 and n-6 polyunsaturated fatty acids, respectively. In the pterygial head, labeling of 4-HHE- and 4-HNE-modified proteins was prominent in the nuclei and cytosol of the epithelium. In the pterygial body, strong labeling was observed in the nuclei and cytosol of the epithelium and proliferating subepithelial connective tissue. In normal conjunctival specimens, only trace immunoreactivity of both proteins was observed in the epithelial and stromal layers. Exposures of ultraviolet (330 nm, 48.32 ± 0.55 J/cm2 or blue light (400 nm, 293.0 ± 2.0 J/cm2 to rat eyes enhanced labeling of 4-HHE- and 4-HNE-modified proteins in the nuclei of conjunctival epithelium. Protein modifications by biologically active aldehydes are a molecular event involved in the development of pterygia.

  3. Protein-modified nanocrystalline diamond thin films for biosensor applications.

    Science.gov (United States)

    Härtl, Andreas; Schmich, Evelyn; Garrido, Jose A; Hernando, Jorge; Catharino, Silvia C R; Walter, Stefan; Feulner, Peter; Kromka, Alexander; Steinmüller, Doris; Stutzmann, Martin

    2004-10-01

    Diamond exhibits several special properties, for example good biocompatibility and a large electrochemical potential window, that make it particularly suitable for biofunctionalization and biosensing. Here we show that proteins can be attached covalently to nanocrystalline diamond thin films. Moreover, we show that, although the biomolecules are immobilized at the surface, they are still fully functional and active. Hydrogen-terminated nanocrystalline diamond films were modified by using a photochemical process to generate a surface layer of amino groups, to which proteins were covalently attached. We used green fluorescent protein to reveal the successful coupling directly. After functionalization of nanocrystalline diamond electrodes with the enzyme catalase, a direct electron transfer between the enzyme's redox centre and the diamond electrode was detected. Moreover, the modified electrode was found to be sensitive to hydrogen peroxide. Because of its dual role as a substrate for biofunctionalization and as an electrode, nanocrystalline diamond is a very promising candidate for future biosensor applications.

  4. Using modified soy protein to enhance foaming of egg white protein.

    Science.gov (United States)

    Wang, Guang; Troendle, Molly; Reitmeier, Cheryll A; Wang, Tong

    2012-08-15

    It is well known that the foaming properties of egg white protein are significantly reduced when a small amount of yolk is mixed in the white. To improve foaming properties of yolk-contaminated egg white protein, soy protein isolate (SPI) and egg proteins were modified to make basic proteins, and effects of these modified proteins on egg white foaming were evaluated in a model and an angel cake system. SPI and egg yolk proteins were modified to have an isoelectric point of 10, and sonication was used to increase protein dispersibility after the ethyl esterification reaction. However, only the addition of sonicated and modified SPI (SMSPI) showed improvement of foaming in the 5% egg protein model system with 0.4% yolk addition. SMSPI was then used in making angel food cake to examine whether the cake performance reduction due to yolk contamination of the white would be restored by such alkaline protein. Cake performance was improved when cream of tartar was used together with SMSPI. Basic soy protein can be made and used to improve egg white foaming properties and cake performance. Copyright © 2012 Society of Chemical Industry.

  5. Competitive Protein Adsorption on Polysaccharide and Hyaluronate Modified Surfaces

    Science.gov (United States)

    Ombelli, Michela; Costello, Lauren; Postle, Corinne; Anantharaman, Vinod; Meng, Qing Cheng; Composto, Russell J.; Eckmann, David M.

    2011-01-01

    We measured adsorption of bovine serum albumin (BSA) and fibrinogen (Fg) onto six distinct bare and dextran- and hyaluronate-modified silicon surfaces created using two dextran grafting densities and three hyaluronic acid (HA) sodium salts derived from human umbilical cord, rooster comb and streptococcus zooepidemicus. Film thickness and surface morphology depended on HA molecular weight and concentration. BSA coverage was enhanced on surfaces upon competitive adsorption of BSA:Fg mixtures. Dextranization differentially reduced protein adsorption onto surfaces based on oxidation state. Hyaluronization was demonstrated to provide the greatest resistance to protein coverage, equivalent to that of the most resistant dextranized surface. Resistance to protein adsorption was independent of the type of hyaluronic acid utilized. With changing bulk protein concentration from 20 to 40 µg ml−1 for each species, Fg coverage on silicon increased by 4×, whereas both BSA and Fg adsorption on dextran and HA were far less dependent of protein bulk concentration. PMID:21623481

  6. Fifty years of radiochemical tracers

    International Nuclear Information System (INIS)

    Evans, E.A.

    1992-01-01

    During the past 50 years radiochemical tracers, usually in the form of isotopically labelled organic compounds, have been essential tools to further advance our knowledge at the frontiers of a great variety of scientific developments in the life sciences. This plenary lecture reviews necessarily selected highlights in the synthesis and applications of such radiochemical tracers. Included are examples where important advances, made possible by using radiochemicals, have contributed to improving the quality of life on this planet. The principal radioisotopes involved, 14 C, 3 H, 35 S, 32 P, 125 I, are all relatively safe to handle and are commercially available at maximum theoretical specific activity (carrier free). The compounds labeled with these radioisotopes are used in many fields of research which include biosynthesis and biotechnology studies, cell biology, drug metabolism, clinical research and environmental applications, and are briefly reviewed. (author). 55 refs

  7. Determination of radiochemical purity using gas chromatography

    International Nuclear Information System (INIS)

    1975-01-01

    The concepts of chromatography, gas chromatography, activity, radiochemical impurity are defined; the procedure of the application of gas chromatography for detecting radiochemical purity of substances is standardized. (E.F.)

  8. Preparation of Modified Films with Protein from Grouper Fish

    Science.gov (United States)

    Tecante, A.; Granados-Navarrete, S.; Martínez-García, C.

    2016-01-01

    A protein concentrate (PC) was obtained from Grouper fish skin and it was used to prepare films with different amounts of sorbitol and glycerol as plasticizers. The best performing films regarding resistance were then modified with various concentrations of CaCl2, CaSO4 (calcium salts), and glucono-δ-lactone (GDL) with the purpose of improving their mechanical and barrier properties. These films were characterized by determining their mechanical properties and permeability to water vapor and oxygen. Formulations with 5% (w/v) protein and 75% sorbitol and 4% (w/v) protein with a mixture of 15% glycerol and 15% sorbitol produced adequate films. Calcium salts and GDL increased the tensile fracture stress but reduced the fracture strain and decreased water vapor permeability compared with control films. The films prepared represent an attractive alternative for being used as food packaging materials. PMID:27597950

  9. Preparation of Modified Films with Protein from Grouper Fish

    Directory of Open Access Journals (Sweden)

    M. A. Valdivia-López

    2016-01-01

    Full Text Available A protein concentrate (PC was obtained from Grouper fish skin and it was used to prepare films with different amounts of sorbitol and glycerol as plasticizers. The best performing films regarding resistance were then modified with various concentrations of CaCl2, CaSO4 (calcium salts, and glucono-δ-lactone (GDL with the purpose of improving their mechanical and barrier properties. These films were characterized by determining their mechanical properties and permeability to water vapor and oxygen. Formulations with 5% (w/v protein and 75% sorbitol and 4% (w/v protein with a mixture of 15% glycerol and 15% sorbitol produced adequate films. Calcium salts and GDL increased the tensile fracture stress but reduced the fracture strain and decreased water vapor permeability compared with control films. The films prepared represent an attractive alternative for being used as food packaging materials.

  10. Oxidatively Modified Proteins in the Serous Subtype of Ovarian Carcinoma

    Directory of Open Access Journals (Sweden)

    Sharifeh Mehrabi

    2014-01-01

    Full Text Available Serous subtype of ovarian cancer is considered to originate from fallopian epithelium mucosa that has been exposed to physiological changes resulting from ovulation. Ovulation influences an increased in inflammation of epithelial ovarian cells as results of constant exposure of cells to ROS. The imbalance between ROS and antioxidant capacities, as well as a disruption of redox signaling, causes a wide range of damage to DNA, proteins, and lipids. This study applied spectrophotometric, dinitrophenylhydrazone (DNPH assay, two-dimensional gel electrophoresis, and Western blot analyses to assess the levels of oxidatively modified proteins in 100 primary serous epithelial ovarian carcinoma and normal/surrounding tissues. These samples were obtained from 56 Caucasian and 44 African-American patients within the age range of 61±10 years. Analyses showed that the levels of reactive protein carbonyl groups increased as stages progressed to malignancy. Additionally, the levels of protein carbonyls in serous ovarian carcinoma among African Americans are 40% (P<0.05 higher relative to Caucasian at similar advanced stages. Results suggest that oxidative stress is involved in the modification of carbonyl protein groups, leading to increased aggressiveness of epithelial ovarian tumors and may contribute to the disease's invasiveness among African Americans.

  11. Radiochemical stability of radiopharmaceutical preparations

    International Nuclear Information System (INIS)

    Martins, Patricia de A.; Silva, Jose L. da; Ramos, Marcelo P.S.; Oliveira, Ideli M. de; Felgueiras, Carlos F.; Herrerias, Rosana; Zapparoli Junior, Carlos L.; Mengatti, Jair; Fukumori, Neuza T.O.; Matsuda, Margareth M.N.

    2011-01-01

    The 'in vitro' stability studies of the radiopharmaceutical preparations are an essential requirement for routine practice in nuclear medicine and are an important parameter for evaluating the quality, safety and efficacy required for the sanitary registration of pharmaceutical products. Several countries have published guidelines for the evaluation of pharmaceutical stability. In Brazil, the stability studies should be conducted according to the Guide for Conducting Stability Studies published in the Resolution-RE n. 1, of 29th July 2005. There are also for radiopharmaceutical products, two specific resolutions: RDC-63 regulates the Good Manufacturing Practices for Radiopharmaceuticals and RDC-64 provides the Registration of Radiopharmaceuticals, both published on the 18th December 2009. The radiopharmaceutical stability is defined as the time during which the radioisotope can be safely used for the intended purpose. The radiochemical stability can be affected by a variety of factors, including storage temperature, amount of radioactivity, radioactive concentration, presence or absence of antioxidants or other stabilizing agents. The radiochemical stability studies must be established under controlled conditions determined by the effective use of the product. The aim of this work was to evaluate the radiochemical stability of labeled molecules with 131 I, 123 I, 153 Sm, 18 F, 51 Cr, 177 Lu and 111 In as well as 67 Ga and 201 Tl radiopharmaceuticals. Radiochemical purity was evaluated after production and in the validity period, with the maximum activity and in the recommended storage conditions. The analyses were carried out by thin-layer silica gel plate, paper chromatography and gel chromatography. The experimental results showed to be in accordance with the specified limits for all the analysed products. (author)

  12. Advanced glycation end product (AGE) modified proteins in tears of diabetic patients.

    Science.gov (United States)

    Zhao, Zhenjun; Liu, Jingfang; Shi, Bingyin; He, Shuixiang; Yao, Xiaoli; Willcox, Mark D P

    2010-08-11

    High glucose level in diabetic patients may lead to advanced glycation end product (AGE) modified proteins. This study investigated AGE modified proteins in tears and compared their levels in diabetic patients (DM) with non-diabetic controls (CTL). Basal tears were collected from DM with (DR) or without (DNR) retinopathy and CTL. Total AGE modified proteins were detected quantitatively by a dot immunobinding assay. The AGE modified proteins were separated in 1D- and 2D-SDS gels and detected by western-blotting. The individual AGE modified proteins were also compared between groups using densitometry. Compared with the CTL group, tear concentrations of AGE modified proteins were significantly elevated in DR and DNR groups. The concentration of AGE modified proteins in diabetic tears were positively correlated with AGE modified hemoglobin (HbA1c) and postprandial blood glucose level (PBG). Western blotting of AGE modified proteins from 1D-SDS gels showed several bands, the major one at around 60 kDa. The intensities of AGE modified protein bands were higher in DM tears than in CTL tears. Western blotting from 2D-SDS gels showed a strongly stained horizontal strip, which corresponded to the major band in 1D-SDS gels. Most of the other AGE modified protein species were within molecular weight of 30-60 kDa, PI 5.2-7.0. Densitometry analysis demonstrated several AGE modified proteins were elevated in DR or DNR tears. Total and some individual AGE modified proteins were elevated in DM tears. AGE modified proteins in tears may be used as biomarkers to diagnose diabetes and/or diabetic retinopathy.

  13. Serum lipids modify periodontal infection - C-reactive protein association.

    Science.gov (United States)

    Haro, Anniina; Saxlin, Tuomas; Suominen, Anna-Liisa; Ylöstalo, Pekka; Leiviskä, Jaana; Tervonen, Tellervo; Knuuttila, Matti

    2012-09-01

    To investigate whether low-grade inflammation-related factors such as serum low-density (LDL-C) and high-density lipoprotein cholesterol (HDL-C) modify the association between periodontal infection and C-reactive protein. This study was based on a subpopulation of the Health 2000 Survey, which consisted of dentate, non-diabetic, non-rheumatic subjects who were 30-49 years old (n = 2710). The extent of periodontal infection was measured by means of the number of teeth with periodontal pocket ≥4 mm and teeth with periodontal pocket ≥6 mm and systemic inflammation using high sensitive C-reactive protein. The extent of periodontal infection was associated with elevated levels of C-reactive protein among those subjects whose HDL-C value was below the median value of 1.3 mmol/l or LDL-C above the median value of 3.4 mmol/l. Among those with HDL-C ≥ 1.3 mmol/l or LDL-C ≤ 3.4 mmol/l, the association between periodontal infection and serum concentrations of C-reactive protein was practically non-existent. This study suggests that the relation of periodontal infection to the systemic inflammatory condition is more complicated than previously presumed. The findings of this study suggest that the possible systemic effect of periodontal infection is dependent on serum lipid composition. © 2012 John Wiley & Sons A/S.

  14. Modified procedure for rapid labelling of low concentrations of bioactive proteins with indium-111

    Energy Technology Data Exchange (ETDEWEB)

    Zoghbi, S S; Neumann, R D; Gottschalk, A

    1985-01-01

    The authors describe the conjugation of DTPA to 100-500 g of protein in concentrations of 0.6-1.0 mg mL utilizing the mixed anhydride method. Free DTPA is removed by minicolumn gel filtration and centrifugation with minimal protein dilution. Radiolabelling process can be monitored by instant thin layer chromatography. Any radiochemical impurity detected can be eliminated either by additional minicolumn filtration of further chelation with more conjugated protein. In citrate buffer at pH 6 with minicolumn gel chromatography the authors prepared In-DTPA-D3 (3.0 Ci g) monoclonal antibody and used it to image hepatocarcinoma in guinea pigs. 13 references, 5 figures, 2 tables.

  15. Radiochemical analysis of chlorine-36

    International Nuclear Information System (INIS)

    Rodriguez, M.; Pina, G.; Lara, E.

    2006-01-01

    The radioactive chlorine isotope, 36 Cl, decays with a half-life of 3x10 5 years by emitting a beta particle (98 %) and by electron capture. The aim of this paper is to propose a radiochemical separation method of 36 Cl from the other beta-gamma emitters present in low and medium radioactive wastes such as spent ion exchange resins and evaporator concentrates, that arise from Nuclear Power Plants and particularly in the wastes that come from decommissioning activities of graphite reactors, in order to provide data for 36 Cl inventory calculations. The separation method proposed is based on an oxidation technique where chlorine is trapped by NaOH. 36 Cl beta emissions are measured by liquid scintillation counting by the dual label technique in order to avoid the contamination produced by 14 C which is also trapped by NaOH and which is the main contaminant present in graphite samples. The sensitivity of this method is sufficient to achieve the needed thresholds for the radiological characterization of the radioactive materials to which this method can be applied. (author)

  16. Collected radiochemical and geochemical procedures

    Energy Technology Data Exchange (ETDEWEB)

    Kleinberg, J [comp.

    1990-05-01

    This revision of LA-1721, 4th Ed., Collected Radiochemical Procedures, reflects the activities of two groups in the Isotope and Nuclear Chemistry Division of the Los Alamos National Laboratory: INC-11, Nuclear and radiochemistry; and INC-7, Isotope Geochemistry. The procedures fall into five categories: I. Separation of Radionuclides from Uranium, Fission-Product Solutions, and Nuclear Debris; II. Separation of Products from Irradiated Targets; III. Preparation of Samples for Mass Spectrometric Analysis; IV. Dissolution Procedures; and V. Geochemical Procedures. With one exception, the first category of procedures is ordered by the positions of the elements in the Periodic Table, with separate parts on the Representative Elements (the A groups); the d-Transition Elements (the B groups and the Transition Triads); and the Lanthanides (Rare Earths) and Actinides (the 4f- and 5f-Transition Elements). The members of Group IIIB-- scandium, yttrium, and lanthanum--are included with the lanthanides, elements they resemble closely in chemistry and with which they occur in nature. The procedures dealing with the isolation of products from irradiated targets are arranged by target element.

  17. Radiochemical purity determination by paper chromatography 2

    International Nuclear Information System (INIS)

    1975-01-01

    The standard relates to the determination of radiochemical impurities in labelled compounds using paper chromatography. The basic terms are given as is the description of procedure and evaluation of chromatograms. (E.S.)

  18. Radiochemistry and radiochemical separations. A current bibliography

    International Nuclear Information System (INIS)

    Bujdoso, E.

    1999-01-01

    A current bibliography for years 1993-1996 with 159 references was compiled on radiochemistry and radiochemical separations based on the INIS Atomindex. The references are arranged in alphabetical order of first authors. (N.T.)

  19. Immobilization of ferrocene-modified SNAP-fusion proteins

    NARCIS (Netherlands)

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

    2013-01-01

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

  20. Characterization and Preparation of Broken Rice Proteins Modified by Proteases

    Directory of Open Access Journals (Sweden)

    Lixia Hou

    2010-01-01

    Full Text Available Broken rice is an underutilized by-product of milling. Proteins prepared from broken rice by treatments with alkaline protease and papain have been characterized with regard to nutritional and functional properties. The protein content and the protein recovery were 56.45 and 75.45 % for alkaline protease treatment, and 65.45 and 46.32 % for papain treatment, respectively. Protease treatment increased the lysine and valine content, leading to a more balanced amino acid profile. Broken rice proteins had high emulsifying capacity, 58.3–71.6 % at neutral pH, and adequate water holding capacity, ranging from 1.96 to 2.93 g/g of proteins. At pH=7.0, the broken rice protein had the highest water holding capacity and the best interfacial activities (emulsifying capacity, emulsifying stability, foaming capacity and foaming stability, which may be the result of the higher solubility at pH=7.0. The interfacial activities increased with the increase in the mass fraction of broken rice proteins. The proteins prepared by the papain treatment had higher water holding capacity (p>0.05, emulsifying capacity (p0.05 than alkaline protease treatment at the same pH or mass fraction. To test the fortification of food products with broken rice proteins, pork sausages containing the proteins were prepared. Higher yield of the sausages was obtained with the increased content of broken rice proteins, in the range of 2.0–9.0 %. The results indicate that broken rice proteins have potential to be used as the protein fortification ingredient for food products.

  1. A modified gelatin zymography technique incorporating total protein normalization.

    Science.gov (United States)

    Raykin, Julia; Snider, Eric; Bheri, Sruti; Mulvihill, John; Ethier, C Ross

    2017-03-15

    Gelatinase zymography is a commonly used laboratory procedure; however, variability in sample loading and concentration reduce the accuracy of quantitative results obtained from this technique. To facilitate normalization of gelatinase activity by loaded protein amount, we developed a protocol using the trihalocompound 2,2,2-trichloroethanol to allow for gelatin zymography and total protein labeling within the same gel. We showed that detected protein levels increased linearly with loading, and describe a loading concentration range over which normalized gelatinase activity was constant. We conclude that in-gel total protein detection is feasible in gelatin zymography and greatly improves comparison of gelatinase activity between samples. Published by Elsevier Inc.

  2. Reprocessing of spent nuclear fuel, Annex 2: Chemical-technology study of the modified 'Purex' process Chemical and radiochemical control analyses; Prerada isluzenog nuklearnog goriva, Prilog 2: Hemijsko tehnolosko ispitivanje modifikovanog 'purex' procesa

    Energy Technology Data Exchange (ETDEWEB)

    Gal, I [Institute of Nuclear Sciences Boris Kidric, Laboratorija za hemiju visoke aktivnosti, Vinca, Beograd (Serbia and Montenegro)

    1964-12-15

    The objective of this task was testing of the modified purex process in the constructed separation cell, and verification of the reliability and efficiency of the process. Extractors used were 1BX, 1BS and 1C. testing was done with syntetic solutions.

  3. Safety assessment for TA-48 radiochemical operations

    International Nuclear Information System (INIS)

    1994-08-01

    The purpose of this report is to document an assessment performed to evaluate the safety of the radiochemical operations conducted at the Los Alamos National Laboratory operations area designated as TA-48. This Safety Assessment for the TA-48 radiochemical operations was prepared to fulfill the requirements of US Department of Energy (DOE) Order 5481.1B, ''Safety Analysis and Review System.'' The area designated as TA-48 is operated by the Chemical Science and Technology (CST) Division and is involved with radiochemical operations associated with nuclear weapons testing, evaluation of samples collected from a variety of environmental sources, and nuclear medicine activities. This report documents a systematic evaluation of the hazards associated with the radiochemical operations that are conducted at TA-48. The accident analyses are limited to evaluation of the expected consequences associated with a few bounding accident scenarios that are selected as part of the hazard analysis. Section 2 of this report presents an executive summary and conclusions, Section 3 presents pertinent information concerning the TA-48 site and surrounding area, Section 4 presents a description of the TA-48 radiochemical operations, and Section 5 presents a description of the individual facilities. Section 6 of the report presents an evaluation of the hazards that are associated with the TA-48 operations and Section 7 presents a detailed analysis of selected accident scenarios

  4. Radiochemical separation of cadmium-109

    International Nuclear Information System (INIS)

    Egamediev, S.; Mukhtarov, A.; Nurbaeva, D.; Rakhmanov, A.

    2006-01-01

    Full text: Cadmium-109 has a half-life of 461.9 days and decays by electron capture to 109 Ag with the emission of 88 keV γ-ray (3.79%) along with the characteristic X-ray from the K level of Ag, with energy of 22.5 keV. This radionuclide has found widespread use as a photon source in x-ray fluorescence analysis devices employed in industry for numerous applications such as the direct determination of gold in ores, the analysis of metals and identification of steels. Other applications range from its use as an electron source for measurement of densities of air-pollution samples, to tracer studies in mushrooms and mice and rats. In the nuclear medicine field there is growing interest in employing 109 Cd in a 109 Cd/ 109mA g generator, as an alternative to other biomedical generators of ultra short-lived gamma emitters. There are several methods for the production of 109 Cd in literature: 1. Bombardment of silver cyclotron target via 109 Ag(d,2n) 109 Cd reaction with 16 MeV deuterons. 2. Bombardment of natural silver target via 109 Ag(p,n) 109 Cd reaction with 14 MeV protons. 3. Proton bombardment of natural indium target with 96 MeV protons. 4. Irradiation of enriched 107 Ag target in high-flux nuclear reactor at neutron flux 2x10 15 n·cm -2 ·s -1 via 107 Ag(n,γ) 108 Ag → 108 Cd (n,γ) 109 Cd reaction. 5. Irradiation of enriched 108 Cd target in nuclear reactor at neutron flux 1x10 14 n·cm -2 ·s -1 via 108 Cd (n,γ) 109 Cd reaction. The production of 109 Cd with proton beam via 109 Ag(p,n) 109 Cd reaction is ideal for the cyclotron U-150, since it is not required the change of the regime for the machine functioning. Because of its relatively long half-life the time required for separation is also not an important factor, but its use as an X-ray source requires a very high radiochemical purity. In the present work we studied two methods for separation of 109 Cd from model solution of silver targets. First method is based on precipitation of silver as

  5. Modulating Protein Adsorption on Oxygen Plasma Modified Polysiloxane Surfaces

    International Nuclear Information System (INIS)

    Marletta, G.

    2006-01-01

    In the present paper we report the study on the adsorption behaviour of three model globular proteins, Human Serum Albumin, Lactoferrin and Egg Chicken Lysozyme onto both unmodified surfaces of a silicon-based polymer and the corresponding plasma treated surfaces. In particular, thin films of hydrophobic polysiloxane (about 90 degree of static water contact angle, WCA) were converted by oxygen plasma treatment at reduced pressure into very hydrophilic phases of SiOx (WCA less than 5 degree). The kinetics of protein adsorption processes were investigated by QCM-D technique, while the chemical structure and topography of the protein adlayer have been studied by Angular resolved-XPS and AFM respectively. It turned out that Albumin and Lysozyme exhibited the opposite preferential adsorption respectively onto the hydrophobic and hydrophilic surfaces, while Lactoferrin did not exhibit significant differences. The observed protein behaviour are discussed both in terms of surface-dependent parameters, including surface free energy and chemical structure, and in terms of protein-dependent parameters, including charge as well as the average molecular orientation in the adlayers. Finally, some examples of differential adsorption behaviour of the investigated proteins are reported onto nanopatterned polysiloxane surfaces consisting of hydrophobic nanopores surrounded by hydrophilic (plasma-treated) matrix and the reverse

  6. RADIOCHEMICAL YIELDS OF GRAFT POLYMERIZATION REACTIONS OF CELLULOSE

    Energy Technology Data Exchange (ETDEWEB)

    Arthur, Jr, J C; Blouin, F A

    1963-12-15

    The preparation of radioinduced graft polymers of cotton cellulose, while retaining the fibrous nature and high molecular weight of the cellulose, depended primarily on the radiochemical yields of cellulose reactions and of graft polymerization reactions. Yields of the initial major molecular changes in cellulosic polymer indicated that, in the case of scission of the molecule and carboxyl group formation, chain reactions were not initiated by radiation; however, in the case of carbonyl group formation chain reactions were initiated but quickly terminated. Generally, experimental procedures, used in graft polymerization reactions, were: simultaneous irradiation reactions, that is, application of monomers or solutions of monomers to cellulose or chemically modified celluloses, then irradiation; and post-irradiation reactions, that is, irradiation of cellulose or chemically modified celluloses, then after removal from the field of radiation, contacting the irradiated cellulose with monomer. Some of the most important factors influencing the radiochemical yields of graft polymerization reactions, of styrene and acrylonitrile onto cellulose were: concentration of monomer in treating solution; solvent; ratio of monomer solution to cellulose; prior chemical modification of cellulose; and absence of oxygen, particularly in post-irradiation reactions. Experimental data are presented, and the direct and indirect effects of Co/sup 60/ gamma radiation on these reactions are discussed. (auth)

  7. Sonochemical synthesis of (3-aminopropyl)triethoxysilane-modified monodispersed silica nanoparticles for protein immobilization

    International Nuclear Information System (INIS)

    Shen, Shou-Cang; Ng, Wai Kiong; Chia, Leonard; Dong, Yuan-Cai; Tan, Reginald B.H.

    2011-01-01

    Graphical abstract: 3-Aminopropyltriethoxysilane modified monodispersed silica nanoparticles were synthesized by rapid sonochemical co-condensation to achieve high capability for protein immobilization. Highlights: → Amino-modified monodispersed silica nanoparticles were synthesized by rapid co-condensation. → Strong positive charge was created by aminopropyl-modification. → Capability for immobilization of negatively charged protein was enhanced. → Electrostatic interaction between proteins and surface contributed to the enhanced adsorption. -- Abstract: 3-Aminopropyltriethoxysilane modified monodispersed silica nanoparticles were synthesized by a rapid sonochemical co-condensation synthesis procedure. The chemical nature of surface organic modifier on the obtained modified silica nanoparticle was characterized by 13 C and 29 Si MAS Nuclear Magnetic Resonance (NMR) spectroscopies, Fourier-transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA)- differential scanning calorimetry (DSC). Due to the strengthened positive surface charge of the silica nanoparticles by the modification with aminopropyl groups, the capability for bovine serum albumin (BSA) adsorption was significantly increased as compared with bare silica nanoparticles. 80 mg/g BSA was adsorbed on modified silica nanoparticles, whereas only 20 mg/g BSA could be loaded on pure silica nanoparticles. The enhanced positive surface charge repelled proteins with net positive charge and the modified silica nanoparticles exhibited negligible adsorption of lysozyme, thus a selective adsorption of proteins could be achieved.

  8. Sonochemical synthesis of (3-aminopropyl)triethoxysilane-modified monodispersed silica nanoparticles for protein immobilization

    Energy Technology Data Exchange (ETDEWEB)

    Shen, Shou-Cang, E-mail: shen_shoucang@ices.a-star.edu.sg [Institute of Chemical and Engineering Sciences, A-STAR (Agency for Science, Technology and Research), 1 Pesek Road, Jurong Island, Singapore 627833 (Singapore); Ng, Wai Kiong; Chia, Leonard; Dong, Yuan-Cai [Institute of Chemical and Engineering Sciences, A-STAR (Agency for Science, Technology and Research), 1 Pesek Road, Jurong Island, Singapore 627833 (Singapore); Tan, Reginald B.H., E-mail: reginald_tan@ices.a-star.edu.sg [Institute of Chemical and Engineering Sciences, A-STAR (Agency for Science, Technology and Research), 1 Pesek Road, Jurong Island, Singapore 627833 (Singapore); Department of Chemical and Biomolecular Engineering, The National University of Singapore, 4 Engineering Drive 4, Singapore 117576 (Singapore)

    2011-10-15

    Graphical abstract: 3-Aminopropyltriethoxysilane modified monodispersed silica nanoparticles were synthesized by rapid sonochemical co-condensation to achieve high capability for protein immobilization. Highlights: {yields} Amino-modified monodispersed silica nanoparticles were synthesized by rapid co-condensation. {yields} Strong positive charge was created by aminopropyl-modification. {yields} Capability for immobilization of negatively charged protein was enhanced. {yields} Electrostatic interaction between proteins and surface contributed to the enhanced adsorption. -- Abstract: 3-Aminopropyltriethoxysilane modified monodispersed silica nanoparticles were synthesized by a rapid sonochemical co-condensation synthesis procedure. The chemical nature of surface organic modifier on the obtained modified silica nanoparticle was characterized by {sup 13}C and {sup 29}Si MAS Nuclear Magnetic Resonance (NMR) spectroscopies, Fourier-transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA)- differential scanning calorimetry (DSC). Due to the strengthened positive surface charge of the silica nanoparticles by the modification with aminopropyl groups, the capability for bovine serum albumin (BSA) adsorption was significantly increased as compared with bare silica nanoparticles. 80 mg/g BSA was adsorbed on modified silica nanoparticles, whereas only 20 mg/g BSA could be loaded on pure silica nanoparticles. The enhanced positive surface charge repelled proteins with net positive charge and the modified silica nanoparticles exhibited negligible adsorption of lysozyme, thus a selective adsorption of proteins could be achieved.

  9. Methods for training radiochemical technicians at ORNL

    International Nuclear Information System (INIS)

    Parrott, J.R.; Nicol, R.G.

    The training of personnel to carry out radiochemical operations at ORNL is a formidable and recurrent task since programs are constantly shifting. It is essential that provisions be made for the routine retraining of these personnel if they are to make effective contributions on a continuing basis. Training methods are described that have emerged as a result of thirty years experience in a variety of radiochemical pilot-plant programs. Emphasis is placed on training programs for technicians for the 233 U Processing Facility since essentially all aspects of radiochemical operations are encountered in this facility. These programs have included operations performed in glove boxes, hot-cell manipulator work handling high-neutron-emitting isotopes, and the entire spectrum of remote solvent extraction operations. (U.S.)

  10. Radiochemical analysis of phosphorus in milk samples

    International Nuclear Information System (INIS)

    Oliveira, R.M. de; Cunha, I.I.L.

    1991-01-01

    The determination of phosphorus in milk samples by thermal neutron activation analysis employing radiochemical separation is described. The radiochemical separation consists of the simultaneous irradiation of samples and standards, dissolution of the milk samples in a perchloric acid and nitric acid mixture, addition of zinc hold-back carrier, precipitation of phosphorus as ammonium phospho molybdate (A.M.P.) and sample counting in a Geiger-Mueller detector. The analysis sources of error were studied and the established method was applied to phosphorus analyses in commercial milk samples. (author)

  11. Chromatographic and traditional albumin isotherms on cellulose: a model for wound protein adsorption on modified cotton

    Science.gov (United States)

    Albumin is the most abundant protein found in healing wounds. Traditional and chromatogrpahic protein isotherms of albumin binding on modified cotton fibers are useful in understanding albumin binding to cellulose wound dressings. An important consideration in the design of cellulosic wound dressin...

  12. Biocontainment of genetically modified organisms by synthetic protein design

    Science.gov (United States)

    Mandell, Daniel J.; Lajoie, Marc J.; Mee, Michael T.; Takeuchi, Ryo; Kuznetsov, Gleb; Norville, Julie E.; Gregg, Christopher J.; Stoddard, Barry L.; Church, George M.

    2015-02-01

    Genetically modified organisms (GMOs) are increasingly deployed at large scales and in open environments. Genetic biocontainment strategies are needed to prevent unintended proliferation of GMOs in natural ecosystems. Existing biocontainment methods are insufficient because they impose evolutionary pressure on the organism to eject the safeguard by spontaneous mutagenesis or horizontal gene transfer, or because they can be circumvented by environmentally available compounds. Here we computationally redesign essential enzymes in the first organism possessing an altered genetic code (Escherichia coli strain C321.ΔA) to confer metabolic dependence on non-standard amino acids for survival. The resulting GMOs cannot metabolically bypass their biocontainment mechanisms using known environmental compounds, and they exhibit unprecedented resistance to evolutionary escape through mutagenesis and horizontal gene transfer. This work provides a foundation for safer GMOs that are isolated from natural ecosystems by a reliance on synthetic metabolites.

  13. Radiochemical Solar Neutrino Experiments - Successful and Otherwise

    International Nuclear Information System (INIS)

    Hahn, R.L.

    2008-01-01

    Over the years, several different radiochemical systems have been proposed as solar neutrino detectors. Of these, two achieved operating status and obtained important results that helped to define the current field of neutrino physics: the first solar-neutrino experiment, the Chlorine Detector ( 37 Cl) that was developed by chemist Raymond Davis and colleagues at the Homestake Mine, and the subsequent Gallium ( 71 Ga) Detectors that were operated by (a) the SAGE collaboration at the Baksan Laboratory and (b) the GALLEX/GNO collaborations at the Gran Sasso National Laboratory. These experiments have been extensively discussed in the literature and in many previous International Neutrino Conferences. In this paper, I present important updates to the results from SAGE and GALLEX/GNO. I also review the principles of the radiochemical detectors and briefly describe several different detectors that have been proposed. In light of the well-known successes that have been subsequently obtained by real-time neutrino detectors such as Kamiokande, Super-Kamiokande, SNO, and KamLAND, I do not anticipate that any new radiochemical neutrino detectors will be built. At present, only SAGE is still operating; the Chlorine and GNO radiochemical detectors have been decommissioned and dismantled

  14. Radiochemical solar neutrino experiments, 'successful and otherwise'

    International Nuclear Information System (INIS)

    Hahn, Richard L

    2008-01-01

    Over the years, several different radiochemical systems have been proposed as solar neutrino detectors. Of these, two achieved operating status and obtained important results that helped to define the current field of neutrino physics: the first solar-neutrino experiment, the Chlorine Detector ( 37 Cl) that was developed by chemist Raymond Davis and colleagues at the Homestake Mine, and the subsequent Gallium ( 71 Ga) Detectors that were operated by (a) the SAGE collaboration at the Baksan Laboratory and (b) the GALLEX/GNO collaborations at the Gran Sasso National Laboratory. These experiments have been extensively discussed in the literature and in many previous International Neutrino Conferences. In this paper, I present important updates to the results from SAGE and GALLEX/GNO. I also review the principles of the radiochemical detectors and briefly describe several different detectors that have been proposed. In light of the well-known successes that have been subsequently obtained by real-time neutrino detectors such as Kamiokande, Super-Kamiokande, SNO, and KamLAND, I do not anticipate that any new radiochemical neutrino detectors will be built. At present, only SAGE is still operating; the Chlorine and GNO radiochemical detectors have been decommissioned and dismantled.

  15. Radiochemical studies on nuclear fission at Trombay

    Indian Academy of Sciences (India)

    227Ac to 245Cm were determined by radiochemical methods which involved ... foil, followed by direct γ counting using high resolution Ge(Li) detector was also ... the stiffness to mass asymmetric distortion decreases on either side of lead.Also ...

  16. Efficiency of Database Search for Identification of Mutated and Modified Proteins via Mass Spectrometry

    OpenAIRE

    Pevzner, Pavel A.; Mulyukov, Zufar; Dancik, Vlado; Tang, Chris L

    2001-01-01

    Although protein identification by matching tandem mass spectra (MS/MS) against protein databases is a widespread tool in mass spectrometry, the question about reliability of such searches remains open. Absence of rigorous significance scores in MS/MS database search makes it difficult to discard random database hits and may lead to erroneous protein identification, particularly in the case of mutated or post-translationally modified peptides. This problem is especially important for high-thr...

  17. Using the Ubiquitin-modified Proteome to Monitor Distinct and Spatially Restricted Protein Homeostasis Dysfunction.

    Science.gov (United States)

    Gendron, Joshua M; Webb, Kristofor; Yang, Bing; Rising, Lisa; Zuzow, Nathan; Bennett, Eric J

    2016-08-01

    Protein homeostasis dysfunction has been implicated in the development and progression of aging related human pathologies. There is a need for the establishment of quantitative methods to evaluate global protein homoeostasis function. As the ubiquitin (ub) proteasome system plays a key role in regulating protein homeostasis, we applied quantitative proteomic methods to evaluate the sensitivity of site-specific ubiquitylation events as markers for protein homeostasis dysfunction. Here, we demonstrate that the ub-modified proteome can exceed the sensitivity of engineered fluorescent reporters as a marker for proteasome dysfunction and can provide unique signatures for distinct proteome challenges which is not possible with engineered reporters. We demonstrate that combining ub-proteomics with subcellular fractionation can effectively separate degradative and regulatory ubiquitylation events on distinct protein populations. Using a recently developed potent inhibitor of the critical protein homeostasis factor p97/VCP, we demonstrate that distinct insults to protein homeostasis function can elicit robust and largely unique alterations to the ub-modified proteome. Taken together, we demonstrate that proteomic approaches to monitor the ub-modified proteome can be used to evaluate global protein homeostasis and can be used to monitor distinct functional outcomes for spatially separated protein populations. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  18. Production of Remedial Proteins through Genetically Modified Bacteria

    Directory of Open Access Journals (Sweden)

    Fatima Tariq

    2018-02-01

    Full Text Available Recombinant DNA technology has created biological organisms with advanced genetic sequences and has been extensively used to express multiple genes for therapeutic purposes when expressed in a suitable host. Microbial systems such as prokaryotic bacteria has been successfully utilized as a heterologous systems showing high therapeutic potency for various human diseases. Bioengineered bacteria have been successfully utilized for producing therapeutic proteins, treating infectious diseases, and disease arise due to increasing resistance to antibiotics. Prominently E. coli found to be the most widely used expression system for recombinant therapeutic protein production i.e. hormones, enzymes and antibodies. Besides E. coli, non-pathogenic lactic acid bacteria has also been considered as an excellent candidate for live mucosal vaccine. Likewise, S. typhimurium has been deployed as attenuated type of vaccination as well as in treatment strategy of various cancers due to its ability of wide progression in tumors. The present article is a summarized view of the main achievements and current developments in the field of recombinant therapeutics using bacterial strains focusing on their usability in therapeutics and future potential.

  19. Functional relevance of G-protein-coupled-receptor-associated proteins, exemplified by receptor-activity-modifying proteins (RAMPs).

    Science.gov (United States)

    Fischer, J A; Muff, R; Born, W

    2002-08-01

    The calcitonin (CT) receptor (CTR) and the CTR-like receptor (CRLR) are close relatives within the type II family of G-protein-coupled receptors, demonstrating sequence identity of 50%. Unlike the interaction between CT and CTR, receptors for the related hormones and neuropeptides amylin, CT-gene-related peptide (CGRP) and adrenomedullin (AM) require one of three accessory receptor-activity-modifying proteins (RAMPs) for ligand recognition. An amylin/CGRP receptor is revealed when CTR is co-expressed with RAMP1. When complexed with RAMP3, CTR interacts with amylin alone. CRLR, initially classed as an orphan receptor, is a CGRP receptor when co-expressed with RAMP1. The same receptor is specific for AM in the presence of RAMP2. Together with human RAMP3, CRLR defines an AM receptor, and with mouse RAMP3 it is a low-affinity CGRP/AM receptor. CTR-RAMP1, antagonized preferentially by salmon CT-(8-32) and not by CGRP-(8-37), and CRLR-RAMP1, antagonized by CGRP-(8-37), are two CGRP receptor isotypes. Thus amylin and CGRP interact specifically with heterodimeric complexes between CTR and RAMP1 or RAMP3, and CGRP and AM interact with complexes between CRLR and RAMP1, RAMP2 or RAMP3.

  20. Effects of insecticidal crystal proteins (Cry proteins) produced by genetically modified maize (Bt maize) on the nematode Caenorhabditis elegans

    International Nuclear Information System (INIS)

    Höss, Sebastian; Menzel, Ralph; Gessler, Frank; Nguyen, Hang T.; Jehle, Johannes A.; Traunspurger, Walter

    2013-01-01

    The genetically modified maize MON89034 × MON88017 expresses different crystal (Cry) proteins with pesticidal activity against the European corn borer (Cry1.105; Cry2Ab2) and the Western corn root worm (Cry3Bb1). Non-target organisms, such as soil nematodes, might be exposed to the Cry proteins that enter the soil in course of crop growing. Therefore, the risk of those proteins for nematodes was assessed by testing their toxic effects on Caenorhabditis elegans. All three insecticidal Cry proteins showed dose-dependent inhibitory effects on C. elegans reproduction (EC50: 0.12–0.38 μmol L −1 ), however, at concentrations that were far above the expected soil concentrations. Moreover, a reduced toxicity was observed when Cry proteins were added jointly. A C. elegans mutant strain deficient for receptors for the nematicidal Cry5B was also resistant against Cry1.105 and Cry2Ab2, suggesting that these Cry proteins bound to the same or similar receptors as nematicidal Cry proteins and thereby affect the reproduction of C. elegans. -- Highlights: •Insecticidal Cry proteins dose-dependently inhibited the reproduction of C. elegans. •Mixture toxicity was lower than expected from concentration-additive single effects. •Genes for MAPK-defense-pathway were up-regulated in presence of Cry protein mixture. •Knock-out strains deficient for Cry5B-receptors showed lower susceptibility to insecticidal Cry proteins. •Toxicity of insecticidal Cry-proteins on C. elegans occurred at concentrations far above expected field concentrations. -- Insecticidal Cry proteins expressed by genetically modified maize act on nematodes via a similar mode of action as nematicidal Cry proteins, however, at concentrations far above expected soil levels

  1. Radiochemical tracers in marine biology

    International Nuclear Information System (INIS)

    Petrocelli, S.R.; Anderson, J.W.; Neff, J.M.

    1977-01-01

    Tracers have been used in a great variety of experimentation. More recently, labeled materials have been applied in marine biological research. Some of the existing tracer techniques have been utilized directly, while others have been modified to suit the specific needs of marine biologists. This chapter describes some of the uses of tracers in marine biological research. It also mentions the problems encountered as well as offering possible solutions and discusses further applications of these techniques. Only pertinent references are cited and additional information may be obtained by consulting these references. Due to their relative ease of maintenance, freshwater species are also utilized in studies which involve radiotracer techniques. Since most of these techniques e directly applicable to marine species, some of these studies will also be included

  2. Methods for training radiochemical technicians at ORNL

    International Nuclear Information System (INIS)

    Parrott, J.R.; Nicol, R.G.

    1975-01-01

    The training of personnel to carry out radiochemical operations at ORNL is a formidable and recurrent task since repetitive, production-type operations are not involved, and programs are constantly shifting. It is essential that provisions be made for the routine retraining of personnel if they are to make effective contributions on a continuing basis. The present training methods have emerged as a result of thirty years experience in a variety of radiochemical pilot-plant programs. These programs have included operations performed in glove boxes, hot-cell manipulator work handling high-neutron-emitting isotopes, and the entire spectrum of remote solvent extraction operations. Present methods of training and the results obtained are summarized

  3. Automated radiochemical processing for clinical PET

    International Nuclear Information System (INIS)

    Padgett, H.C.; Schmidt, D.G.; Bida, G.T.; Wieland, B.W.; Pekrul, E.; Kingsbury, W.G.

    1991-01-01

    With the recent emergence of positron emission tomography (PET) as a viable clinical tool, there is a need for a convenient, cost-effective source of the positron emitter-labeled radiotracers labeled with carbon-11, nitrogen-13, oxygen-15, and fluorine-18. These short-lived radioisotopes are accelerator produced and thus, require a cyclotron and radiochemistry processing instrumentation that can be operated 3 in a clinical environment by competant technicians. The basic goal is to ensure safety and reliability while setting new standards for economy and ease of operation. The Siemens Radioisotope Delivery System (RDS 112) is a fully automated system dedicated to the production and delivery of positron-emitter labeled precursors and radiochemicals required to support a clinical PET imaging program. Thus, the entire RDS can be thought of as an automated radiochemical processing apparatus

  4. Radiochemical Means of Investigating Delayed Neutron Precursors

    International Nuclear Information System (INIS)

    Marmol, P. del

    1968-01-01

    Fast radiochemical methods used now for the determination of delayed neutron precursors are classified and reviewed: precipitations, solvent extractions, range experiments, milking, gas sweeping, isotopic and ion exchange, hot atom reactions and diffusion loss. Advantages and limitations of irradiation systems with respect to fast separations are discussed: external beams which allow faster separations only have low neutron fluxes, internal beams which are mostly fit for gaseous reactions; and rabbits for solution irradiations. Future prospects of radiochemical procedures are presented; among these, studies should be mostly oriented towards gaseous reactions which offer possibilities of isolating very short-lived delayed neutron precursors. Chemical procedures for delayed neutron precursor detection are compared with mass spectrometric and isotope separator techniques; it is concluded that the methods are complementary. (author)

  5. Radiochemical Means of Investigating Delayed Neutron Precursors

    International Nuclear Information System (INIS)

    Marmol, P. del

    1968-01-01

    Fast radiochemical methods used now for the determination of delayed neutron precursors are classified and reviewed: precipitations, solvent extractions, range experiments, milking, gas sweeping, isotopic and ion exchange, hot-atom reactions and diffusion loss. Advantages and limitations of irradiation systems with respect to fast separations are discussed: external beams which allow faster separations only have low neutron fluxes, internal beams which are mostly fit for gaseous reactions; and rabbits for solution irradiations. Future prospects of radiochemical procedures are presented; among these, studies should be mostly oriented towards gaseous reactions which offer possibilities of isolating very short-lived delayed neutron precursors. Chemical procedures for delayed neutron precursor detection are compared with mass spectrometric and isotope-separator techniques; it is concluded that the methods are complementary. (author)

  6. Radiochemical analysis of the Bikini ashes

    Energy Technology Data Exchange (ETDEWEB)

    Ishibashi, M; Shigematsu, T; Ishida, T

    1954-01-01

    The following nuclides were detected in the Bikini ashes by radiochemical procedures: /sup 45/Ca, /sup 89/Sr, /sup 91/Y, /sup 95/Zr, /sup 103/Ru, /sup 144/Pr, and /sup 237/U. The ion-exchange method was used for analysis of contaminated rain water which fell on the Kyoto area on May 16, 1954 from which the presence of /sup 89/Sr, /sup 95/Zr, and /sup 140/Ba, was detected. Rare earths seemed also to be present.

  7. Imaging Scanner Usage in Radiochemical Purity Test

    International Nuclear Information System (INIS)

    Norhafizah Othman; Yahaya Talib; Wan Hamirul Bahrin Wan Kamal

    2011-01-01

    Imaging Scanner model BIOSCAN AR-2000 has been used in the radiochemical purity test for the product of Mo-99/ Tc-99m generator. Result from this test was produced directly where the percentage of pertechnetate was calculated based on width peak area by thin layer chromatography. This paperwork will explain the function, procedure, calibration of the instrument and discussed the advantages compared to the previous method. (author)

  8. Radiochemical analysis of military nuclear facilities

    International Nuclear Information System (INIS)

    Bayramov, A.A.; Bayramova, S.M.

    2012-01-01

    Full text : Radiochemical Analysis is a branch of analytical chemistry comprising an aggregate of methods for qualitatively determining the composition and content of radioisotopes in the products of transformations. Safety and minimization of radiation impact on human and environment are important demand of operation of Military Nuclear Facilities (MNF). In accordance of recommendations of International Commission on Radiological Protection there are next objects of radiochemical analysis: 1) potential sources of radiochemical pollution; 2) environment (objects of environment, human environment including buildings, agricultural production, water, air et al.); 3) human himself (determination of dose from external and internal radiation, chemical poisoning). The chemical analysis can be carried out using, for example, the Gas Chromatography instrument whish separates chemical mixtures and identifies the components at a molecular level. It is one of the most accurate tools for analyzing environmental samples. The Gas Chromatography works on the principle that a mixture will separate into individual substances when heated. The heated gases are carried through a column with an inert gas (such as helium). As the separated substances emerge from the column opening, they flow into the Mass Spectrometry. Mass spectrometry identifies compounds by the mass of the analyte molecule. Newly developed portable Gas Chromatography and Mass Spectrometry are techniques that can be used to separate volatile organic compounds and pesticides. Other uses of Gas Chromatography, combined with other separation and analytical techniques, have been developed for radionuclides, explosive compounds such as royal demolition explosive and trinitrotoluene, and metals. So, based on the many years experience of operation of dangerous MNF, in concordance with norms of radiation and chemical safety it was considered that the tasks of the radiochemical analysis of Military Nuclear Facilities include

  9. 14th radiochemical conference. Booklet of abstracts

    International Nuclear Information System (INIS)

    2002-03-01

    The contributions dealt with the following topics: Radionuclides in the environment, radioecology; Nuclear analytical methods; Chemistry of actinide and trans-actinide elements; Ionizing radiation in science, technology, and arts and cultural heritage preservation; Production and application of radionuclides; Separation methods, speciation; Chemistry of nuclear fuel cycle, radiochemical problems in nuclear waste management; and Nuclear methods in medicine, radiopharmaceuticals, and radiodiagnostics, labelled compounds. Of the verbal and poster presentation, 192 have been input to INIS. (P.A.)

  10. Rapid Radiochemical Methods for Asphalt Paving Material ...

    Science.gov (United States)

    Technical Brief Validated rapid radiochemical methods for alpha and beta emitters in solid matrices that are commonly encountered in urban environments were previously unavailable for public use by responding laboratories. A lack of tested rapid methods would delay the quick determination of contamination levels and the assessment of acceptable site-specific exposure levels. Of special concern are matrices with rough and porous surfaces, which allow the movement of radioactive material deep into the building material making it difficult to detect. This research focuses on methods that address preparation, radiochemical separation, and analysis of asphalt paving materials and asphalt roofing shingles. These matrices, common to outdoor environments, challenge the capability and capacity of very experienced radiochemistry laboratories. Generally, routine sample preparation and dissolution techniques produce liquid samples (representative of the original sample material) that can be processed using available radiochemical methods. The asphalt materials are especially difficult because they do not readily lend themselves to these routine sample preparation and dissolution techniques. The HSRP and ORIA coordinate radiological reference laboratory priorities and activities in conjunction with HSRP’s Partner Process. As part of the collaboration, the HSRP worked with ORIA to publish rapid radioanalytical methods for selected radionuclides in building material matrice

  11. The movement protein and coat protein of alfalfa mosaic virus accumulate in structurally modified plasmodesmata

    NARCIS (Netherlands)

    van der Wel, N. N.; Goldbach, R. W.; van Lent, J. W.

    1998-01-01

    In systemically infected tissues of Nicotiana benthamiana, alfalfa mosaic virus (AMV) coat protein (CP) and movement protein (MP) are detected in plasmodesmata in a layer of three to four cells at the progressing front of infection. Besides the presence of these viral proteins, the plasmodesmata are

  12. Highly Efficient Intracellular Protein Delivery by Cationic Polyethyleneimine-Modified Gelatin Nanoparticles

    Directory of Open Access Journals (Sweden)

    Ming-Ju Chou

    2018-02-01

    Full Text Available Intracellular protein delivery may provide a safe and non-genome integrated strategy for targeting abnormal or specific cells for applications in cell reprogramming therapy. Thus, highly efficient intracellular functional protein delivery would be beneficial for protein drug discovery. In this study, we generated a cationic polyethyleneimine (PEI-modified gelatin nanoparticle and evaluated its intracellular protein delivery ability in vitro and in vivo. The experimental results showed that the PEI-modified gelatin nanoparticle had a zeta potential of approximately +60 mV and the particle size was approximately 135 nm. The particle was stable at different biological pH values and temperatures and high protein loading efficiency was observed. The fluorescent image results revealed that large numbers of particles were taken up into the mammalian cells and escaped from the endosomes into the cytoplasm. In a mouse C26 cell-xenograft cancer model, particles accumulated in cancer cells. In conclusion, the PEI-modified gelatin particle may provide a biodegradable and highly efficient protein delivery system for use in regenerative medicine and cancer therapy.

  13. Multilayer Choline Phosphate Molecule Modified Surface with Enhanced Cell Adhesion but Resistance to Protein Adsorption.

    Science.gov (United States)

    Chen, Xingyu; Yang, Ming; Liu, Botao; Li, Zhiqiang; Tan, Hong; Li, Jianshu

    2017-08-22

    Choline phosphate (CP), which is a new zwitterionic molecule, and has the reverse order of phosphate choline (PC) and could bind to the cell membrane though the unique CP-PC interaction. Here we modified a glass surface with multilayer CP molecules using surface-initiated atom-transfer radical polymerization (SI-ATRP) and the ring-opening method. Polymeric brushes of (dimethylamino)ethyl methacrylate (DMAEMA) were synthesized by SI-ATRP from the glass surface. Then the grafted PDMAEMA brushes were used to introduce CP groups to fabricate the multilayer CP molecule modified surface. The protein adsorption experiment and cell culture test were used to evaluate the biocompatibility of the modified surfaces by using human umbilical veinendothelial cells (HUVECs). The protein adsorption results demonstrated that the multilayer CP molecule decorated surface could prevent the adsorption of fibrinogen and serum protein. The adhesion and proliferation of cells were improved significantly on the multilayer CP molecule modified surface. Therefore, the biocompatibility of the material surface could be improved by the modified multilayer CP molecule, which exhibits great potential for biomedical applications, e.g., scaffolds in tissue engineering.

  14. A modified FASP protocol for high-throughput preparation of protein samples for mass spectrometry.

    Directory of Open Access Journals (Sweden)

    Jeremy Potriquet

    Full Text Available To facilitate high-throughput proteomic analyses we have developed a modified FASP protocol which improves the rate at which protein samples can be processed prior to mass spectrometry. Adapting the original FASP protocol to a 96-well format necessitates extended spin times for buffer exchange due to the low centrifugation speeds tolerated by these devices. However, by using 96-well plates with a more robust polyethersulfone molecular weight cutoff membrane, instead of the cellulose membranes typically used in these devices, we could use isopropanol as a wetting agent, decreasing spin times required for buffer exchange from an hour to 30 minutes. In a typical work flow used in our laboratory this equates to a reduction of 3 hours per plate, providing processing times similar to FASP for the processing of up to 96 samples per plate. To test whether our modified protocol produced similar results to FASP and other FASP-like protocols we compared the performance of our modified protocol to the original FASP and the more recently described eFASP and MStern-blot. We show that all FASP-like methods, including our modified protocol, display similar performance in terms of proteins identified and reproducibility. Our results show that our modified FASP protocol is an efficient method for the high-throughput processing of protein samples for mass spectral analysis.

  15. Pyrenebutanoate as a dynamic protein modifier for fluorometric detection in capillary zone electrophoresis

    Czech Academy of Sciences Publication Activity Database

    Horká, Marie; Šlais, Karel

    2002-01-01

    Roč. 23, 7-8 (2002), s. 1090-1095 ISSN 0173-0835 R&D Projects: GA AV ČR IAA4031901 Institutional research plan: CEZ:AV0Z4031919 Keywords : pyrenebutanoate * dynamic protein modifier * CZE Subject RIV: CB - Analytical Chemistry, Separation Impact factor: 4.325, year: 2002

  16. Imaging the lipidome: omega-alkynyl fatty acids for detection and cellular visualization of lipid-modified proteins.

    Science.gov (United States)

    Hannoush, Rami N; Arenas-Ramirez, Natalia

    2009-07-17

    Fatty acylation or lipid modification of proteins controls their cellular activation and diverse roles in physiology. It mediates protein-protein and protein-membrane interactions and plays an important role in regulating cellular signaling pathways. Currently, there is need for visualizing lipid modifications of proteins in cells. Herein we report novel chemical probes based on omega-alkynyl fatty acids for biochemical detection and cellular imaging of lipid-modified proteins. Our study shows that omega-alkynyl fatty acids of varying chain length are metabolically incorporated onto cellular proteins. Using fluorescence imaging, we describe the subcellular distribution of lipid-modified proteins across a panel of different mammalian cell lines and during cell division. Our results demonstrate that this methodology is a useful diagnostic tool for analyzing the lipid content of cellular proteins and for studying the dynamic behavior of lipid-modified proteins in various disease or physiological states.

  17. Synthesis and structural characterization of carboxyethylpyrrole-modified proteins: mediators of age-related macular degeneration.

    Science.gov (United States)

    Lu, Liang; Gu, Xiaorong; Hong, Li; Laird, James; Jaffe, Keeve; Choi, Jaewoo; Crabb, John; Salomon, Robert G

    2009-11-01

    Protein modifications in which the epsilon-amino group of lysyl residues is incorporated into a 2-(omega-carboxyethyl)pyrrole (CEP) are mediators of age-related macular degeneration (AMD). They promote both angiogenesis into the retina ('wet AMD') and geographic retinal atrophy ('dry AMD'). Blood levels of CEPs are biomarkers for clinical prognosis of the disease. To enable mechanistic studies of their role in promoting AMD, for example, through the activation of B- and T-cells, interaction with receptors, or binding with complement proteins, we developed an efficient synthesis of CEP derivatives, that is especially effective for proteins. The structures of tryptic peptides derived from CEP-modified proteins were also determined. A key finding is that 4,7-dioxoheptanoic acid 9-fluorenylmethyl ester reacts with primary amines to provide 9-fluorenylmethyl esters of CEP-modified proteins that can be deprotected in situ with 1,8-diazabicyclo[5.4.0]undec-7-ene without causing protein denaturation. The introduction of multiple CEP-modifications with a wide variety of CEP:protein ratios is readily achieved using this strategy.

  18. Application of radiochemical determination methods in cleanability research of building materials

    International Nuclear Information System (INIS)

    Maeaettae, Jenni; Kymaelaeinen, Hanna-Riitta; Sjoeberg, Anna-Maija

    2011-01-01

    During recent years increasing effort has been made to modify surface properties with easy-to-clean or self-cleaning characteristics, and concomitantly there is a need to be able to quantify cleanability. Methodology is a complex issue, including aspects of selection and characterization of the surface materials, the soiling materials (contaminants), soiling and cleaning methods, and the detection methods. Different biological, chemical, physical and visual methods have been included in studies of surface cleanability. One challenge has been to obtain quantitative information about soiling. The radiochemical methods, gamma spectrometry (NaI(Tl)-crystal) and liquid scintillation counting, have been shown to be suitable for evaluating cleanability of different surface materials and different soiling material types, providing quantitative information about the amount of soiling material both on and beneath the surface. Due to the different labelled soiling components, the interaction of the surface with different soiling material types can be evaluated. Radiochemical methods have unique benefits particularly for examining porous materials and surfaces. However, they are suitable only for highly controlled studies because of the hazards. Different features and details of radiochemical methods are discussed with the view to aid planning of future cleanability studies. - Highlights: → Radiochemical methods can be used for cleanability studies. → These methods give quantitative information about the amount of soiling material. → These methods are suitable particularly for examining porous materials. → These methods are suitable for highly controlled studies because of the hazards.

  19. Brown pigment formation in heated sugar-protein mixed suspensions containing unmodified and peptically modified whey protein concentrates.

    Science.gov (United States)

    Rongsirikul, Narumol; Hongsprabhas, Parichat

    2016-01-01

    Commercial whey protein concentrate (WPC) was modified by heating the acidified protein suspensions (pH 2.0) at 80 °C for 30 min and treating with pepsin at 37 °C for 60 min. Prior to spray-drying, such modification did not change the molecular weights (MWs) of whey proteins determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). After spray-drying the modified whey protein concentrate with trehalose excipient (MWPC-TH), it was found that the α-lactalbumin (α-La) was the major protein that was further hydrolyzed the most. The reconstituted MWPC-TH contained β-lactoglobulin (β-Lg) as the major protein and small molecular weight (MW) peptides of less than 6.5 kDa. The reconstituted MWPC-TH had higher NH2 group, Trolox equivalent antioxidant capacity (TEAC), lower exposed aromatic ring and thiol (SH) contents than did the commercial WPC. Kinetic studies revealed that the addition of MWPC-TH in fructose-glycine solution was able to reduce brown pigment formation in the mixtures heated at 80 to 95 °C by increasing the activation energy (Ea) of brown pigment formation due to the retardation of fluoresced advanced glycation end product (AGEs) formation. The addition of MWPC to reducing sugar-glycine/commercial WPC was also able to lower brown pigment formation in the sterilized (121 °C, 15 min) mixed suspensions containing 0.1 M reducing sugar and 0.5-1.0 % glycine and/or commercial (P < 0.05). It was demonstrated that the modification investigated in this study selectively hydrolyzed α-La and retained β-Lg for the production of antibrowning whey protein concentrate.

  20. Potential allergenicity research of Cry1C protein from genetically modified rice.

    Science.gov (United States)

    Cao, Sishuo; He, Xiaoyun; Xu, Wentao; Luo, Yunbo; Ran, Wenjun; Liang, Lixing; Dai, Yunqing; Huang, Kunlun

    2012-07-01

    With the development of genetically modified crops, there has been a growing interest in available approaches to assess the potential allergenicity of novel gene products. We were not sure whether Cry1C could induce allergy. We examined the protein with three other proteins to determine the potential allergenicity of Cry1C protein from genetically modified rice. Female Brown Norway (BN) rats received 0.1 mg peanut agglutinin (PNA), 1mg potato acid phosphatase (PAP), 1mg ovalbumin (OVA) or 5 mg purified Cry1C protein dissolved in 1 mL water by daily gavage for 42 days to test potential allergenicity. Ten days after the last gavage, rats were orally challenged with antigens, and physiologic and immunologic responses were studied. In contrast to sensitization with PNA, PAP and OVA Cry1C protein did not induce antigen-specific IgG2a in BN rats. Cytokine expression, serum IgE and histamine levels and the number of eosinophils and mast cells in the blood of Cry1C group rats were comparable to the control group rats, which were treated with water alone. As Cry1C did not show any allergenicity, we make the following conclusion that the protein could be safety used in rice or other plants. Copyright © 2012 Elsevier Inc. All rights reserved.

  1. Diaphragms obtained by radiochemical grafting in PTFE

    International Nuclear Information System (INIS)

    Nenner, T.; Fahrasmane, A.

    1984-01-01

    Diaphragms for alkaline water electrolysis are prepared by radiochemical grafting of PTFE fabric with styrene, which is later on sulfonated, or with acrylic acid. The diaphragms obtained are mechanically resistant to potash at temperatures up to 200 0 C, but show some degrafting, which limits the lifetime. The sulfonated styrene group has been found to be more stable in electrolysis than the acrylic acid. In both cases, the incorporation of a cross-linking agent like divinyl benzene improves the lifetime of the diaphragms. Electrolysis during 500 hours at 120 0 C and 10 kAm 2 could be performed. (author)

  2. 13th Radiochemical Conference. Booklet of Abstracts

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1998-04-01

    The Conference included the following sessions: (i) Opening plenary presentations (6 contributions); (ii) Chemistry of natural radionuclides, discovery of radium and polonium (6 verbal presentations + 5 poster presentations); (iii) Radionuclides in the environment, radioecology (29 + 48); (iv) Activation analysis and other radioanalytical methods (36 + 49); (v) Ionizing radiation in science and technology (12 + 12); (vi) Chemistry of actinide and trans-actinide elements (11 + 14); (vii) Separation methods, speciation (18 + 41); (viii) Production and application of radionuclides (14 + 29); and (ix) Radiochemical problems in nuclear waste management (12 + 22). The majority of verbal presentations has been input to INIS, mostly in the form of the full authors` abstracts. (P.A.)

  3. 13th Radiochemical Conference. Booklet of Abstracts

    International Nuclear Information System (INIS)

    1998-04-01

    The Conference included the following sessions: (i) Opening plenary presentations (6 contributions); (ii) Chemistry of natural radionuclides, discovery of radium and polonium (6 verbal presentations + 5 poster presentations); (iii) Radionuclides in the environment, radioecology (29 + 48); (iv) Activation analysis and other radioanalytical methods (36 + 49); (v) Ionizing radiation in science and technology (12 + 12); (vi) Chemistry of actinide and trans-actinide elements (11 + 14); (vii) Separation methods, speciation (18 + 41); (viii) Production and application of radionuclides (14 + 29); and (ix) Radiochemical problems in nuclear waste management (12 + 22). The majority of verbal presentations has been input to INIS, mostly in the form of the full authors' abstracts. (P.A.)

  4. Hanford radiochemical site decommissioning demonstration program

    International Nuclear Information System (INIS)

    Nelson, D.C.

    1971-01-01

    A program is proposed for the innovation, development, and demonstration of technologies necessary to decommission the Hanford radiochemical plant area to the extent that the sites can have unrestricted public access. The five tasks selected for development and demonstration of restoration techniques were restoration of a burial ground, decommissioning of a separations plant, restoration of a separations plant waste interim storage tank farm, restoration of a liquid disposal area, and disposal of large contaminated equipment. Process development requirements are tabulated and discussed. A proposed schedule and estimated costs are given

  5. DNA-modified electrodes fabricated using copper-free click chemistry for enhanced protein detection.

    Science.gov (United States)

    Furst, Ariel L; Hill, Michael G; Barton, Jacqueline K

    2013-12-31

    A method of DNA monolayer formation has been developed using copper-free click chemistry that yields enhanced surface homogeneity and enables variation in the amount of DNA assembled; extremely low-density DNA monolayers, with as little as 5% of the monolayer being DNA, have been formed. These DNA-modified electrodes (DMEs) were characterized visually, with AFM, and electrochemically, and were found to facilitate DNA-mediated reduction of a distally bound redox probe. These low-density monolayers were found to be more homogeneous than traditional thiol-modified DNA monolayers, with greater helix accessibility through an increased surface area-to-volume ratio. Protein binding efficiency of the transcriptional activator TATA-binding protein (TBP) was also investigated on these surfaces and compared to that on DNA monolayers formed with standard thiol-modified DNA. Our low-density monolayers were found to be extremely sensitive to TBP binding, with a signal decrease in excess of 75% for 150 nM protein. This protein was detectable at 4 nM, on the order of its dissociation constant, with our low-density monolayers. The improved DNA helix accessibility and sensitivity of our low-density DNA monolayers to TBP binding reflects the general utility of this method of DNA monolayer formation for DNA-based electrochemical sensor development.

  6. Statistical processing of technological and radiochemical data

    International Nuclear Information System (INIS)

    Lahodova, Zdena; Vonkova, Kateřina

    2011-01-01

    The project described in this article had two goals. The main goal was to compare technological and radiochemical data from two units of nuclear power plant. The other goal was to check the collection, organization and interpretation of routinely measured data. Monitoring of analytical and radiochemical data is a very valuable source of knowledge for some processes in the primary circuit. Exploratory analysis of one-dimensional data was performed to estimate location and variability and to find extreme values, data trends, distribution, autocorrelation etc. This process allowed for the cleaning and completion of raw data. Then multiple analyses such as multiple comparisons, multiple correlation, variance analysis, and so on were performed. Measured data was organized into a data matrix. The results and graphs such as Box plots, Mahalanobis distance, Biplot, Correlation, and Trend graphs are presented in this article as statistical analysis tools. Tables of data were replaced with graphs because graphs condense large amounts of information into easy-to-understand formats. The significant conclusion of this work is that the collection and comprehension of data is a very substantial part of statistical processing. With well-prepared and well-understood data, its accurate evaluation is possible. Cooperation between the technicians who collect data and the statistician who processes it is also very important. (author)

  7. Simple protein structure-sensitive chronopotentiometric analysis with dithiothreitol-modified Hg electrodes

    Czech Academy of Sciences Publication Activity Database

    Ostatná, Veronika; Černocká, Hana; Paleček, Emil

    2012-01-01

    Roč. 87, SI (2012), s. 84-88 ISSN 1567-5394 R&D Projects: GA AV ČR(CZ) KJB100040901; GA ČR(CZ) GAP301/11/2055; GA MŠk(CZ) LC06035 Institutional research plan: CEZ:AV0Z50040507; CEZ:AV0Z50040702 Keywords : protein electroanalysis * DTT-modified electrodes * electrocatalysis Subject RIV: BO - Biophysics Impact factor: 3.947, year: 2012

  8. Comparison of Zirconium Phosphonate-Modified Surfaces for Immobilizing Phosphopeptides and Phosphate-Tagged Proteins.

    Science.gov (United States)

    Forato, Florian; Liu, Hao; Benoit, Roland; Fayon, Franck; Charlier, Cathy; Fateh, Amina; Defontaine, Alain; Tellier, Charles; Talham, Daniel R; Queffélec, Clémence; Bujoli, Bruno

    2016-06-07

    Different routes for preparing zirconium phosphonate-modified surfaces for immobilizing biomolecular probes are compared. Two chemical-modification approaches were explored to form self-assembled monolayers on commercially available primary amine-functionalized slides, and the resulting surfaces were compared to well-characterized zirconium phosphonate monolayer-modified supports prepared using Langmuir-Blodgett methods. When using POCl3 as the amine phosphorylating agent followed by treatment with zirconyl chloride, the result was not a zirconium-phosphonate monolayer, as commonly assumed in the literature, but rather the process gives adsorbed zirconium oxide/hydroxide species and to a lower extent adsorbed zirconium phosphate and/or phosphonate. Reactions giving rise to these products were modeled in homogeneous-phase studies. Nevertheless, each of the three modified surfaces effectively immobilized phosphopeptides and phosphopeptide tags fused to an affinity protein. Unexpectedly, the zirconium oxide/hydroxide modified surface, formed by treating the amine-coated slides with POCl3/Zr(4+), afforded better immobilization of the peptides and proteins and efficient capture of their targets.

  9. ZZ HATCHES-18, Database for radiochemical modelling

    International Nuclear Information System (INIS)

    Heath, T.G.

    2008-01-01

    1 - Description of program or function: HATCHES is a referenced, quality assured, thermodynamic database, developed by Serco Assurance for Nirex. Although originally compiled for use in radiochemical modelling work, HATCHES also includes data suitable for many other applications e.g. toxic waste disposal, effluent treatment and chemical processing. It is used in conjunction with chemical and geochemical computer programs, to simulate a wide variety of reactions in aqueous environments. The database includes thermodynamic data (the log formation constant and the enthalpy of formation for the chemical species) for the actinides, fission products and decay products. The datasets for Ni, Tc, U, Np, Pu and Am are based on the NEA reviews of the chemical thermodynamics of these elements. The data sets for these elements with oxalate, citrate and EDTA are based on the NEA-selected values. For iso-saccharinic acid, additional data (non-selected values) have been included from the NEA review as well as data derived from other sources. HATCHES also includes data for many toxic metals and for elements commonly found in groundwaters or geological materials. HARPHRQ operates by reference to the PHREEQE master species list. Thus the thermodynamic information supplied is: a) the log equilibrium constant for the formation reaction of the requested species from the PHREEQE master species for the corresponding elements; b) the enthalpy of reaction for the formation reaction of the requested species from the PHREEQE master species for the corresponding elements. This version of HATCHES has been updated since the previous release to provide consistency with the selected data from two recent publications in the OECD Nuclear Energy Agency series on chemical thermodynamics: Chemical Thermodynamics Series Volume 7 (2005): Chemical Thermodynamics of Selenium by Aeke Olin (Chairman), Bengt Nolaeng, Lars-Olof Oehman, Evgeniy Osadchii and Erik Rosen and Chemical Thermodynamics Series Volume 8

  10. Isolation of pronephros cells which endocytose chemically modified proteins in the rainbow trout

    International Nuclear Information System (INIS)

    Dannevig, B.H.; Berg, T.

    1986-01-01

    Modified serum albumin is cleared from the blood by kidney cells in salmonid fishes. The present study deals with isolation of cells from pronephros which endocytose formaldehyde-treated human serum albumin (fHSA). Radioactively labelled fHSA or dinitrophenyl-conjugated albumin (DNP-HSA) were injected intravenously into rainbow trouts. Pronephros cells, containing the endocytosed protein, were isolated and further separated by centrifugal elutriation and density-gradient centrifugation. Most of the radioactive protein was elutriated together with small cells. After centrifuging the cells through a Percoll density gradient, radioactive protein was located in cells recovered in the upper part of the gradient. In mammals, fHSA and other modified proteins are mainly taken up by sinusoidal endothelial cells in the liver via a scavenger receptor 0. Our results suggest that a comparable function in salmonids is located in a subpopulation of relatively small cells in kidney tissue, possibly sinusoidal lining cells. The separation techniques used seemed to be suitable for isolation of different populations of pronephros cells

  11. Cleanout and decontamination of radiochemical hot cells

    International Nuclear Information System (INIS)

    Surma, J.E.; Holton, L.K. Jr.; Katayama, Y.B.; Gose, J.E.; Haun, F.E.; Dierks, R.D.

    1990-01-01

    The Pacific Northwest Laboratory is developing and employing advanced remote and contact technologies in cleaning out and decontaminating six radiochemical hot cells at Hanford under the Department of Energy's Surplus Facilities Management Program. The program is using a series of remote and contact decontamination techniques to reduce costs and to significantly lower radiation doses to workers. Refurbishment of the cover blocks above the air lock trench reduced radiation exposure in the air lock and cleanout and decontamination of an analytical cell achieved a reduction in radioactive contamination. Nuclear Regulatory Commission-approved Type B burial boxes are also being used to reduce waste disposal costs and radiation doses. PNL is currently decommissioning its pilot-scale radioactive liquid-fed ceramic melter. Special tools have been developed and are being used to accomplish the world's first such effort. 4 refs., 5 figs

  12. Mixing and sampling tests for Radiochemical Plant

    International Nuclear Information System (INIS)

    Ehinger, M.N.; Marfin, H.R.; Hunt, B.

    1999-01-01

    The paper describes results and test procedures used to evaluate uncertainly and basis effects introduced by the sampler systems of a radiochemical plant, and similar parameters associated with mixing. This report will concentrate on experiences at the Barnwell Nuclear Fuels Plant. Mixing and sampling tests can be conducted to establish the statistical parameters for those activities related to overall measurement uncertainties. Density measurements by state-of-the art, commercially availability equipment is the key to conducting those tests. Experience in the U.S. suggests the statistical contribution of mixing and sampling can be controlled to less than 0.01 % and with new equipment and new tests in operating facilities might be controlled to better accuracy [ru

  13. Radiochemical Analysis Methodology for uranium Depletion Measurements

    Energy Technology Data Exchange (ETDEWEB)

    Scatena-Wachel DE

    2007-01-09

    This report provides sufficient material for a test sponsor with little or no radiochemistry background to understand and follow physics irradiation test program execution. Most irradiation test programs employ similar techniques and the general details provided here can be applied to the analysis of other irradiated sample types. Aspects of program management directly affecting analysis quality are also provided. This report is not an in-depth treatise on the vast field of radiochemical analysis techniques and related topics such as quality control. Instrumental technology is a very fast growing field and dramatic improvements are made each year, thus the instrumentation described in this report is no longer cutting edge technology. Much of the background material is still applicable and useful for the analysis of older experiments and also for subcontractors who still retain the older instrumentation.

  14. Standardization of equations for radiochemical calculations

    International Nuclear Information System (INIS)

    Danahy, R.J.; Dugan, T.A.; Tomlinson, F.K.; Jones, H.W.

    1994-01-01

    In mid 1993, the Fernald Environmental Restoration Management Corporation (FERMCO), with USEPA approval implemented a project quality assurance plan containing performance-based specifications for radiochemical sample analyses conducted in support of the Fernald site remediation activities. FERMCO's initial approach to acquiring performance-based radioanalytical services was to provide limited guidance regarding equations for computation of the quantities required in each analysis report. It became evident that there was a significant divergence of opinion on how to compute some very basic radiochemical quantities. The use of a standardized set of equations was needed in order to ensure comparability of data from different laboratories. In a remediation project of this magnitude, use of multiple laboratories is a virtual necessity. Consequently comparability of data becomes an extremely important issue. A critical issue in the Remedial Investigation/Feasibility Study (RI/FS) phase of the dean up project is to avoid the occurrence of excessive false positive sample results. Such results could lead to unnecessary clean up and significant additional cost. This paper describes the specific formulas FERMCO is currently using to define such quantities as net sample count rate, sample radionuclide concentration, radiometric tracer and gravimetric carrier recovery. Equations have also been produced to define the uncertainty in each of the above quantities. Equations for the Total Propagated Uncertainty (TPU) and for a sample-specific Minimum Detectable Concentration (MDC) have also been specified. Generalized equations have been reformulated to address the specific conditions which apply to the analysis of FERMCO samples. In particular, FERMCO requires results which have been corrected for the radioactivity in the blank while in other instances, sample results without blank correction are required

  15. Characterisation of chemically-modified proteins by electrospray ionisation mass spectrometry

    International Nuclear Information System (INIS)

    Bennett, K.L.

    1996-09-01

    Electrospray mass spectrometry (ESI-MS) has been used to examine a range of intact monoclonal antibodies (MAbs), antibody fragments such as F(ab') 2 , F ab and F c , chemically-modified fragments and a range of other chemically-modified peptides and proteins as part of a broader study aimed at establishing ESI-MS as a method for the characterisation of radioimmunoconjugates (radiolabelled monoclonal antibodies). For example, the addition of up to 10 biotin molecules to the 'papain-sensitive' 50 kDa F ab fragment can be easily detected in ESI mass spectra. For intact MAbs, however, it is only possible to detect average shifts in the mass of intact antibodies following modification. Successful ESI-MS analysis of complexes formed between chelators and other small molecules conjugated to synthetic peptides, hen egg-white Iysozyme (HEL) (M r 14 306) and horse heart myoglobin (M r 16 951) has been demonstrated. ESI-MS offers considerable advantages compared with existing methods for the characterisation of chemically-conjugated proteins including speed and sensitivity of analysis and the capability for obtaining specific structural information. The conditions for ESI-MS of intact MAbs and MAb fragments have been examined in detail and it was found that 150 kDa MAbs generally required lower sample concentration and higher skimmer potentials compared with the 50 kDa F ab fragment and other lower molecular weight proteins. In addition, the m/z range over which ions from MAbs were observed was higher (m/z ∼2000-4500) than for smaller proteins. ESI-MS was also found to be useful for probing the action of the protease papain, that is used to generate MAb fragments (F(ab) '2, F ab and F c ). Further, different sensitivities to papain for different MAb preparations was demonstrated. Finally, the tandem mass spectra of a range of peptides modified by iodine and biotin were examined. In the case of biotinylated peptides, a characteristic fragment ion was identified that could

  16. Protein arrangement on modified diamond-like carbon surfaces – An ARXPS study

    Energy Technology Data Exchange (ETDEWEB)

    Oosterbeek, Reece N., E-mail: reece.oosterbeek@auckland.ac.nz [Department of Chemical and Materials Engineering, The University of Auckland, Private Bag 92019 (New Zealand); Seal, Christopher K. [Light Metals Research Centre, The University of Auckland, Private Bag 92019 (New Zealand); Hyland, Margaret M. [Department of Chemical and Materials Engineering, The University of Auckland, Private Bag 92019 (New Zealand)

    2014-12-01

    Highlights: • DLC coatings were modified by Ar{sup +} ion sputtering and laser graphitisation. • The surface properties of the coatings were measured, and it was found that the above methods increased sp{sup 2} content and altered surface energy. • ARXPS was used to observe protein arrangement on the surface. • Polar CO/CN groups were seen to be segregated towards the interface, indicating they play an important role in bonding. • This segregation increased with increasing polar surface energy, indicating an increased net attraction between polar groups. - Abstract: Understanding the nature of the interface between a biomaterial implant and the biological fluid is an essential step towards creating improved implant materials. This study examined a diamond-like carbon coating biomaterial, the surface energy of which was modified by Ar{sup +} ion sputtering and laser graphitisation. The arrangement of proteins was analysed by angle resolved X-ray photoelectron spectroscopy, and the effects of the polar component of surface energy on this arrangement were observed. It was seen that polar groups (such as CN, CO) are more attracted to the coating surface due to the stronger polar interactions. This results in a segregation of these groups to the DLC–protein interface; at increasing takeoff angle (further from to DLC–protein interface) fewer of these polar groups are seen. Correspondingly, groups that interact mainly by dispersive forces (CC, CH) were found to increase in intensity as takeoff angle increased, indicating they are segregated away from the DLC–protein interface. The magnitude of the segregation was seen to increase with increasing polar surface energy, this was attributed to an increased net attraction between the solid surface and polar groups at higher polar surface energy (γ{sub S}{sup p})

  17. Protein arrangement on modified diamond-like carbon surfaces – An ARXPS study

    International Nuclear Information System (INIS)

    Oosterbeek, Reece N.; Seal, Christopher K.; Hyland, Margaret M.

    2014-01-01

    Highlights: • DLC coatings were modified by Ar + ion sputtering and laser graphitisation. • The surface properties of the coatings were measured, and it was found that the above methods increased sp 2 content and altered surface energy. • ARXPS was used to observe protein arrangement on the surface. • Polar CO/CN groups were seen to be segregated towards the interface, indicating they play an important role in bonding. • This segregation increased with increasing polar surface energy, indicating an increased net attraction between polar groups. - Abstract: Understanding the nature of the interface between a biomaterial implant and the biological fluid is an essential step towards creating improved implant materials. This study examined a diamond-like carbon coating biomaterial, the surface energy of which was modified by Ar + ion sputtering and laser graphitisation. The arrangement of proteins was analysed by angle resolved X-ray photoelectron spectroscopy, and the effects of the polar component of surface energy on this arrangement were observed. It was seen that polar groups (such as CN, CO) are more attracted to the coating surface due to the stronger polar interactions. This results in a segregation of these groups to the DLC–protein interface; at increasing takeoff angle (further from to DLC–protein interface) fewer of these polar groups are seen. Correspondingly, groups that interact mainly by dispersive forces (CC, CH) were found to increase in intensity as takeoff angle increased, indicating they are segregated away from the DLC–protein interface. The magnitude of the segregation was seen to increase with increasing polar surface energy, this was attributed to an increased net attraction between the solid surface and polar groups at higher polar surface energy (γ S p )

  18. Protein classification using modified n-grams and skip-grams.

    Science.gov (United States)

    Islam, S M Ashiqul; Heil, Benjamin J; Kearney, Christopher Michel; Baker, Erich J

    2018-05-01

    Classification by supervised machine learning greatly facilitates the annotation of protein characteristics from their primary sequence. However, the feature generation step in this process requires detailed knowledge of attributes used to classify the proteins. Lack of this knowledge risks the selection of irrelevant features, resulting in a faulty model. In this study, we introduce a supervised protein classification method with a novel means of automating the work-intensive feature generation step via a Natural Language Processing (NLP)-dependent model, using a modified combination of n-grams and skip-grams (m-NGSG). A meta-comparison of cross-validation accuracy with twelve training datasets from nine different published studies demonstrates a consistent increase in accuracy of m-NGSG when compared to contemporary classification and feature generation models. We expect this model to accelerate the classification of proteins from primary sequence data and increase the accessibility of protein characteristic prediction to a broader range of scientists. m-NGSG is freely available at Bitbucket: https://bitbucket.org/sm_islam/mngsg/src. A web server is available at watson.ecs.baylor.edu/ngsg. erich_baker@baylor.edu. Supplementary data are available at Bioinformatics online.

  19. Hot-spot analysis to dissect the functional protein-protein interface of a tRNA-modifying enzyme.

    Science.gov (United States)

    Jakobi, Stephan; Nguyen, Tran Xuan Phong; Debaene, François; Metz, Alexander; Sanglier-Cianférani, Sarah; Reuter, Klaus; Klebe, Gerhard

    2014-10-01

    Interference with protein-protein interactions of interfaces larger than 1500 Ų by small drug-like molecules is notoriously difficult, particularly if targeting homodimers. The tRNA modifying enzyme Tgt is only functionally active as a homodimer. Thus, blocking Tgt dimerization is a promising strategy for drug therapy as this protein is key to the development of Shigellosis. Our goal was to identify hot-spot residues which, upon mutation, result in a predominantly monomeric state of Tgt. The detailed understanding of the spatial location and stability contribution of the individual interaction hot-spot residues and the plasticity of motifs involved in the interface formation is a crucial prerequisite for the rational identification of drug-like inhibitors addressing the respective dimerization interface. Using computational analyses, we identified hot-spot residues that contribute particularly to dimer stability: a cluster of hydrophobic and aromatic residues as well as several salt bridges. This in silico prediction led to the identification of a promising double mutant, which was validated experimentally. Native nano-ESI mass spectrometry showed that the dimerization of the suggested mutant is largely prevented resulting in a predominantly monomeric state. Crystal structure analysis and enzyme kinetics of the mutant variant further support the evidence for enhanced monomerization and provide first insights into the structural consequences of the dimer destabilization. © 2014 Wiley Periodicals, Inc.

  20. Evaluation of a modified IRMA for anti-D quantitation, using 3H protein A

    International Nuclear Information System (INIS)

    Dumasia, A.; Gupte, S.

    1993-01-01

    A modified immunoradiometric assay (IRMA) using tritiated ( 3 H) protein A was developed to estimate anti-D concentration. The main advantages of the assay were longer shelf life of the labelled reagent (more than two years); minimum radiation hazard and; low non specific binding. Levels of anti-D were estimated in 23 Rh (D) immunized women. A good correlation of anti-D concentration (μg/ml) with Rh antibody titre was observed (r=+ 0.89, P 3 H protein A IRMA correlated well with the severity of Rh-HDN. This assay could quantitate anti-D in sera having exclusively IgG 3 subtype. (author). 20 refs., 2 figs., 2 tabs

  1. Lactose repressor protein modified with dansyl chloride: activity effects and fluorescence properties

    International Nuclear Information System (INIS)

    Hsieh, W.T.; Matthews, K.S.

    1985-01-01

    Chemical modification using 5-(dimethylamino)naphthalene-1-sulfonyl chloride (dansyl chloride) has been used to explore the importance of lysine residues involved in the binding activities of the lactose repressor and to introduce a fluorescent probe into the protein. Dansyl chloride modification of lac repressor resulted in loss of operator DNA binding at low molar ratios of reagent/monomer. Loss of nonspecific DNA binding was observed only at higher molar ratios, while isopropyl beta-D-thiogalactoside binding was not affected at any of the reagent levels studied. Lysine residues were the only modified amino acids detected. Protection of lysines-33 and -37 from modification by the presence of nonspecific DNA correlated with maintenance of operator DNA binding activity, and reaction of lysine-37 paralleled operator binding activity loss. Energy transfer between dansyl incorporated in the core region of the repressor protein and tryptophan-201 was observed, with an approximate distance of 23 A calculated between these two moieties

  2. A thiophene-modified screen printed electrode for detection of dengue virus NS1 protein.

    Science.gov (United States)

    Silva, M M S; Dias, A C M S; Cordeiro, M T; Marques, E; Goulart, M O F; Dutra, R F

    2014-10-01

    A thiophene-modified screen printed electrode (SPE) for detection of the Dengue virus non-structural protein 1 (NS1), an important marker for acute phase diagnosis, is described. A sulfur-containing heterocyclic compound, the thiophene was incorporated to a carbon ink to prepare reproducible screen printed electrodes. After cured, the thiophene SPE was coated by gold nanoparticles conjugated to Protein A to form a nanostrutured surface. The Anti-NS1 antibodies immobilized via their Fc portions via Protein A, leaving their antigen specific sites free circumventing the problem of a random antibodies immobilization. Amperometric responses to the NS1 protein of dengue virus were obtained by cyclic voltammetries performed in presence of ferrocyanide/ferricyanide as redox probe. The calibration curve of immunosensor showed a linear response from 0.04 µg mL(-1) to 0.6 µg mL(-1) of NS1 with a good linear correlation (r=0.991, pink enhanced the electroanalytical properties of the SPEs, increasing their reproducibility and sensitivity. This point-of-care testing represents a great potential for use in epidemic situations, facilitating the early diagnosis in acute phase of dengue virus. Copyright © 2014 Elsevier B.V. All rights reserved.

  3. Protein arrangement on modified diamond-like carbon surfaces - An ARXPS study

    Science.gov (United States)

    Oosterbeek, Reece N.; Seal, Christopher K.; Hyland, Margaret M.

    2014-12-01

    Understanding the nature of the interface between a biomaterial implant and the biological fluid is an essential step towards creating improved implant materials. This study examined a diamond-like carbon coating biomaterial, the surface energy of which was modified by Ar+ ion sputtering and laser graphitisation. The arrangement of proteins was analysed by angle resolved X-ray photoelectron spectroscopy, and the effects of the polar component of surface energy on this arrangement were observed. It was seen that polar groups (such as CN, CO) are more attracted to the coating surface due to the stronger polar interactions. This results in a segregation of these groups to the DLC-protein interface; at increasing takeoff angle (further from to DLC-protein interface) fewer of these polar groups are seen. Correspondingly, groups that interact mainly by dispersive forces (CC, CH) were found to increase in intensity as takeoff angle increased, indicating they are segregated away from the DLC-protein interface. The magnitude of the segregation was seen to increase with increasing polar surface energy, this was attributed to an increased net attraction between the solid surface and polar groups at higher polar surface energy (γSp).

  4. Chemically modified carbon nanotubes as material enhanced laser desorption ionisation (MELDI) material in protein profiling

    International Nuclear Information System (INIS)

    Najam-ul-Haq, M.; Rainer, M.; Schwarzenauer, T.; Huck, C.W.; Bonn, G.K.

    2006-01-01

    Biomarkers play a potential role in the early detection and diagnosis of a disease. Our aim is to derivatize carbon nanotubes for exploration of the differences in human body fluids e.g. serum, through matrix assisted laser desorption ionisation/time of flight mass spectrometry (MALDI/TOF-MS) that can be related to disease and subsequently to be employed in the biomarker discovery process. This application we termed as the material enhanced laser desorption ionisation (MELDI). The versatility of this technology is meant to increase the amount of information from biological samples on the protein level, which will have a major impact to serve the cause of diagnostic markers. Serum peptides and proteins are immobilized on derivatized carbon nanotubes, which function as binding material. Protein-loaded suspension is placed on a stainless steel target or buckypaper on aluminum target for direct analysis with MALDI-MS. The elution method to wash the bound proteins from carbon nanotubes was employed to compare with the direct analysis procedure. Elution is carried out by MALDI matrix solution to get them out of the entangled nanotubes, which are difficult to desorb by laser due to the complex nanotube structures. The advantage of these optimized methods compared to the conventional screening methods is the improved sensitivity, selectivity and the short analysis time without prior albumin and immunoglobulin depletion. The comparison of similarly modified diamond and carbon nanotubes exhibit differences in their nature to bind the proteins out of serum due to the differences in their physical characteristics. Infrared (IR) spectroscopy provided hint for the presence of tertiary amine peak at the crucial chemical step of iminodiacetic acid addition to acid chloride functionality on carbon nanotubes. Atomic absorption spectroscopy (AAS) was utilized to quantitatively measure the copper capacity of these derivatized carbon nanotubes which is a direct measure of capacity of

  5. Mesoporous silica particles modified with graphitic carbon: interaction with human red blood cells and plasma proteins

    Energy Technology Data Exchange (ETDEWEB)

    Martinez, Diego Stefani Teodoro; Franqui, Lidiane Silva; Bettini, Jefferson; Strauss, Mathias, E-mail: diego.martinez@lnnano.cnpem.br [Centro Nacional de Pesquisa em Energia e Materiais (CNPEM), Campinas, SP (Brazil); Damasceno, Joao Paulo Vita; Mazali, Italo Odone [Universidade Estadual de Campinas (UNICAMP), SP (Brazil)

    2016-07-01

    Full text: In this work the interaction of the mesoporous silica particles (SBA-15, ∼700 nm) modified with graphitic carbon (SBA-15/C) on human red blood cells (hemolysis) and plasma proteins (protein corona formation) is studied. XPS and CHN analysis showed that the carbon content on the SBA-15/C samples varied from 2 to 10% and was tuned by the functionalization step. The formed carbon structures where associated to graphitic nanodomains coating the pores surface as verified by Raman spectroscopy and {sup 13}C NMR. Advanced TEM/EELS analysis showed that the carbon structures are distributed along the SBA-15 mesopores. SAXS and textural analyses were used to confirm that the porous structure of the silica support is kept after the modification procedure and to calculate the number of graphitic carbon stacked layers coating the mesopores. After incubation of SBA-15 with human red blood cells (RBCs), it was observed a dose-dependent hemolytic effect, probably, due to binding of the material silanol-rich surface to the phosphatidylcholine molecules from the RBC membrane. The graphitic carbon modifications have mitigated this effect, indicating that the graphitic carbon coating protected the silanol groups of the particle surface hindering the hemolysis. Considering the protein corona formation, selective biomolecular interaction of proteins was observed for the different materials using gel electrophoresis (SDS-PAGE) analysis. Besides, graphitic carbon modification decreased the amount of proteins on the corona. Together, the in vitro hemolysis and protein corona assays are promising biological models to understand the influence of silica surface functionalization on their bionano-interactions. Finally, our work contributes to the development of fundamental research on such nanomaterials chemistry in the emerging field of nanobioscience and nanotoxicology. (author)

  6. Novel Highly Sensitive Protein Sensors Based on Tapered Optical Fibres Modified with Au-Based Nanocoatings

    Directory of Open Access Journals (Sweden)

    Aitor Urrutia

    2016-01-01

    Full Text Available Novel protein sensors based on tapered optical fibres modified with Au coatings deposited using two different procedures are proposed. Au-based coatings are deposited onto a nonadiabatic tapered optical fibre using (i a novel facile method composed of layer-by-layer deposition consisting of polycation (poly(allylamine hydrochloride, PAH and negatively charged SiO2 nanoparticles (NPs followed by the deposition of the charged Au NPs and (ii the sputtering technique. The Au NPs and Au thin film surfaces are then modified with biotin in order to bind streptavidin (SV molecules and detect them. The sensing principle is based on the sensitivity of the transmission spectrum of the device to changes in the refractive index of the coatings induced by the SV binding to the biotin. Both sensors showed high sensitivity to SV, with the lowest measured concentration levels below 2.5 nM. The calculated binding constant for the biotin-SV pair was 2.2×10-11 M−1 when a tapered fibre modified with the LbL method was used, with a limit of detection (LoD of 271 pM. The sensor formed using sputtering had a binding constant of 1.01×10-10 M−1 with a LoD of 806 pM. These new structures and their simple fabrication technique could be used to develop other biosensors.

  7. Agrobacterium-mediated transformation of modified antifreeze protein gene in strawberry

    Directory of Open Access Journals (Sweden)

    Srisulak Dheeranupattana

    2005-07-01

    Full Text Available The optimum condition for shoot regeneration from leaf explants of strawberry cultivar Tiogar was investigated. It was found that the best regeneration condition was MS medium containing N6-Benzyladenine (BA and 2,4-Dichlorophenoxy acetic acid (2,4-D at concentrations of 1 mg.l-1 and 0.2 mg.l-1, respectively. Antibiotics sensitivity test found that shoot regeneration from leaf explant was inhibited more than 90% at the concentration of kanamycin (Km as low as 5 mg.l-1. The modified gene encoding antifreeze protein isoform HPLC 6 was successfully constructed using codons which were optimally expressed in the strawberry plant. The antifreeze protein genes, naturally in plasmid pSW1 and modified in plasmid BB, were transformed to strawberry leaf explants by Agrobacterium tumefaciens LBA 4404. The strawberry plants, transformed with both AFP genes, were able to root in MS media containing 50 mg.l-1 Km, while no roots grew from nontransformed plant in this condition. Polymerase chain reaction indicated that the transgenes were integrated in the genome of transformants.

  8. Heterologous protein secretion in Lactobacilli with modified pSIP vectors.

    Directory of Open Access Journals (Sweden)

    Ingrid Lea Karlskås

    Full Text Available We describe new variants of the modular pSIP-vectors for inducible gene expression and protein secretion in lactobacilli. The basic functionality of the pSIP system was tested in Lactobacillus strains representing 14 species using pSIP411, which harbors the broad-host-range Lactococcus lactis SH71rep replicon and a β-glucuronidase encoding reporter gene. In 10 species, the inducible gene expression system was functional. Based on these results, three pSIP vectors with different signal peptides were modified by replacing their narrow-host-range L. plantarum 256rep replicon with SH71rep and transformed into strains of five different species of Lactobacillus. All recombinant strains secreted the target protein NucA, albeit with varying production levels and secretion efficiencies. The Lp_3050 derived signal peptide generally resulted in the highest levels of secreted NucA. These modified pSIP vectors are useful tools for engineering a wide variety of Lactobacillus species.

  9. Amperometric Immunosensor Based on a Protein A/Deposited Gold Nanocrystals Modified Electrode for Carbofuran Detection

    Directory of Open Access Journals (Sweden)

    Xia Sun

    2011-12-01

    Full Text Available In this paper, an amperometric immunosensor modified with protein A/deposited gold nanocrystals (DpAu was developed for the ultrasensitive detection of carbofuran residues. First, DpAu were electrodeposited onto the Au electrode surface to absorb protein A (PA and improve the electrode conductivity. Then PA was dropped onto the surface of DpAu film, used for binding antibody Fc fragments. Next, anti-carbofuran monoclonal antibody was immobilized on the PA modified electrode. Finally, bovine serum albumin (BSA was employed to block the possible remaining active sites avoiding any nonspecific adsorption. The fabrication procedure of the immunosensor was characterized by electrochemical impedance spectroscopy (EIS and cyclic voltammetry (CV, respectively. With the excellent electroconductivity of DpAu and the PA’s oriented immobilization of antibodies, a highly efficient immuno-reaction and detection sensitivity could be achieved. The influences of the electrodeposition time of DpAu, pH of the detection solution and incubation time on the current response of the fabricated immunosensor were investigated. Under optimized conditions, the current response was proportional to the concentration of carbofuran which ranged from 1 to 100 ng/mL and 100 ng/mL to 100 μg/mL. The detection limit was 0.1924 ng/mL. The proposed carbofuran immnuosensor exhibited high specificity, reproducibility, stability and regeneration performance, which may open a new door for ultrasensitive detection of carbofuran residues in vegetables and fruits.

  10. Radiochemical surveillance of KNK primary sodium

    International Nuclear Information System (INIS)

    Stamm, H.-H.; Stade, K.Ch.

    1987-05-01

    Radiochemical surveillance of the KNK primary sodium has been performed now for 15 years with 953 effective full-power days. The overflow method used for sodium sampling proved to be reliable. Different crucible materials have been used for different analytical tasks. The amount of radionuclides in the primary system has not given restrictions to plant operation at any time. On-line gamma spectroscopy on pipings and components of the primary circuits was accomplished in reactor downtimes. Activity depositions on the walls were dominated by Ta-182 after KNK I operation. Main deposited activities at KNK II were Mn-54 (fresh core) and after operation with failed fuel Cs-137, in cover gas areas together with Zn-65. Efficient experimental radionuclide traps for the removal of Mn-54, Zn-65 and Cs-137 from the primary coolant were tested successfully. The dose rates on primary pipes and components of KNK I and KNK II were lower by an order of magnitude compared to water-cooled reactors. This is in good agreement with experiences from LMFBR's in other countries. The resulting average yearly accumulated personal dose rate was 0.21 man-Sv at KNK, compared to 3.9 man-Sv at German light-water-cooled power reactors

  11. Radiochemical studies on amorphous zirconium phosphate

    Energy Technology Data Exchange (ETDEWEB)

    Dyer, A; Moores, G E [Salford Univ. (UK). Dept. of Chemistry and Applied Chemistry

    1981-01-01

    Amorphous zirconium phosphate (ZrP) is used in some hemodialysis machines for the regeneration of dialysate. Its function is to adsorb ammonium ions formed by the pretreatment of urea by urease. It also adsorbs Ca, Mg and K ions but leaches phosphate ions which are then removed (along with F/sup -/ ions) by a bed of hydrous zirconium oxide. The sodium form of ZrP is used although other forms have been suggested for use. The work reported here describes some preliminary radiochemical studies on the mechanism of release of phosphate ions and its possible relationship to sodium ion-exchange. /sup 32/P labelled material (HHZrP) was used for elution experiments with deionized water and buffer solutions having the pH's 4.2, 7.0 and 9.2. Buffer solutions used were as supplied by BDH. Elution was at four different temperatures in the range 293 to 363/sup 0/C. In the second series of experiments HHZrP was suspended in a NaCl solution labelled with /sup 22/Na. From this, /sup 22/Na labelled ZrP (NaHZrP) was prepared and eluted in the same way as the HHZrP. Results are given and discussed.

  12. Radiochemical measurement of mass transport in sodium

    International Nuclear Information System (INIS)

    Cooper, M.H.; Chiang, S.H.

    1976-01-01

    Mass transport processes in the sodium coolant of Liquid Metal Fast Breeder Reactors (LMFBRs) are significant in determining rates of corrosion and deposition of radioactive nuclides from the fuel cladding, deposition and cold trapping of fission products from defect or failed fuel, carbon and nitrogen redistribution in the containment materials, and removal of impurities by cold trapping or hot trapping. Mass transport between rotating, concentric cylinders in molten sodium has been investigated using a unique radiochemical method. Long-lived (33 year) cesium-137, dissolved in the sodium, decays radioactively emitting a beta to barium-137m, which decays with a short half-life (2.6 minutes) emitting a gamma. Cesium is weakly adsorbed and remains in solution, while the barium is strongly adsorbed on the stainless steel surfaces. Hence, by measuring the barium-137m activity on movable stainless steel surfaces, one can calculate the mass transport to that surface. Mass transfer coefficients in sodium measured by this method are in agreement with published heat transfer correlations when the effect of the volumetric mass source is taken into account. Hence, heat transfer correlations can be confidently utilized by analogy in estimating mass transfer in liquid-metal systems

  13. New radiochemical methods for determination of 237Np a 241Pu using extraction chromatography (Presentation)

    International Nuclear Information System (INIS)

    Strisovska, J.

    2013-01-01

    Thesis was focused on the development of a new methodology for the separation of anthropogenic transuranium radionuclides 237 Np a 241 Pu from different kinds of matrices. The analytical methods used in this study were based on extraction chromatography and were optimized according to the sample type. The proposed radiochemical procedure is a combination of two algorithms, which represent the separation of radionuclides by using extraction chromatographic sorbents TEVA resin and TRU resin supplied by Eichrom Technologies LLC. 239 Np a 237 Np were selectively captured on sorbent TEVA resin in oxidation state 4+. TRU resin was used for purification of plutonium fraction from interfering americium radionuclide. 242 Pu and 239 Np radionuclides as tracers have been used to monitor the radiochemical yields of separation. Before every radiochemical separation tracer radionuclide 239 Np was obtained by separation from the parent radionuclide 2 43 Am, which is in radioactive equilibrium to 239 Np. The average yield of chemical separation was 69,3% for 239 Np at 277 keV energy line and 65,9% at 228 keV energy line. The NPL AH-B08069 (2008) samples which consist of the mixture of alpha-radionuclides were used for the modification and optimization of separation method used for separation of Np and Pu in model samples. This method provided high radiochemical yields of 239,240 Pu (95,0 ± 3,5)% and 237 Np (87,9 ± 3,0)%.. Reliability of the method was verified by applying our modified separation procedures on reference materials IAEA-375 and IAEA-414 supplied by International Atomic Energy Agency. A good agreement between the results is obtained by this procedure and the certified values were found. Samples of contaminated soils from the area of Nuclear power plant A-1 Jaslovske Bohunice which is stored temporarily before disposal were analyzed using developed separation procedure. Specific activity of investigated radionuclides was determined in these samples. (author)

  14. The use of robots for automation in the radiochemical laboratory

    International Nuclear Information System (INIS)

    Huddleston, J.

    1988-01-01

    The use of robotic systems for automated processes such as overnight operations, procedures involving radiation hazards in radiochemical laboratories is discussed. Particular reference is made to their use in analytical problems. Their flexibility is emphasised. (U.K.)

  15. A radiochemical assay for biotin in biological materials

    International Nuclear Information System (INIS)

    Hood, R.L.

    1975-01-01

    A radiochemical assay for biotin is described. The assay was sensitive to one nanogram and simple enough for routine biotin analyses. The assay yielded results which were comparable to those obtained from a microbiological assay using Lactobacillus plantarum. (author)

  16. The publication lapse of papers in Radiochemical and Radioanalytical Letters

    International Nuclear Information System (INIS)

    Braun, T.; Nagydiosi-Kocsis, Gy.

    1982-01-01

    The time needed for passing through journal editorial and publication processing has been examined for the papers published in Radiochemical and Radioanalytical Letters for the years 1969-1981. (author)

  17. Safety and Waste Management for SAM Radiochemical Methods

    Science.gov (United States)

    The General Safety and Waste Management page offers section-specific safety and waste management details for the radiochemical analytes included in EPA's Selected Analytical Methods for Environmental Remediation and Recovery (SAM).

  18. Pharmacological characterization of receptor-activity-modifying proteins (RAMPs) and the human calcitonin receptor.

    Science.gov (United States)

    Armour, S L; Foord, S; Kenakin, T; Chen, W J

    1999-12-01

    Receptor-activity-modifying proteins (RAMPs) are a family of single transmembrane domain proteins shown to be important for the transport and ligand specificity of the calcitonin gene-related peptide (CGRP) receptor. In this report, we describe the analysis of pharmacological properties of the human calcitonin receptor (hCTR) coexpressed with different RAMPs with the use of the Xenopus laevis melanophore expression system. We show that coexpression of RAMP3 with human calcitonin receptor changed the relative potency of hCTR to human calcitonin (hCAL) and rat amylin. RAMP1 and RAMP2, in contrast, had little effect on the change of hCTR potency to hCAL or rat amylin. When coexpressed with RAMP3, hCTR reversed the relative potency by a 3.5-fold loss in sensitivity to hCAL and a 19-fold increase in sensitivity to rat amylin. AC66, an inverse agonist, produced apparent simple competitive antagonism of hCAL and rat amylin, as indicated by linear Schild regressions. The potency of AC66 was changed in the blockade of rat amylin but not hCAL responses with RAMP3 coexpression. The mean pK(B) for AC66 to hCAL was 9.4 +/- 0.3 without RAMP3 and 9.45 +/- 0.07 with RAMP3. For the antagonism of AC66 to rat amylin, the pK(B) was 9.25 +/- 0.15 without RAMP3 and 8.2 +/- 0.35 with RAMP3. The finding suggests that RAMP3 might modify the active states of calcitonin receptor in such a way as to create a new receptor phenotype that is "amylin-like." Irrespective of the physiological association of the new receptor species, the finding that a coexpressed membrane protein can completely change agonist and antagonist affinities for a receptor raises implications for screening in recombinant receptor systems.

  19. Radiochemical and instrumental neutron activation analysis - recent trends

    International Nuclear Information System (INIS)

    Dams, R.

    1990-01-01

    Recent trends of radiochemical and instrumental neutron activation analysis are discussed. Novel developments include the application of cyclic and pulsed activation, better energy resolution with hyperpure germanium detectors, and use of pulse processing systems allowing extremely high count rates of very short-lived isotopes. Further development is anticipated in the field of speciation in biological and environmental studies. Radiochemical methods have led to accurate determinations at the ng/g level. A promising future is expected for neutron activation techniques. (orig.)

  20. Adhesion of Trypanosoma cruzi trypomastigotes to fibronectin or laminin modifies tubulin and paraflagellar rod protein phosphorylation.

    Directory of Open Access Journals (Sweden)

    Eliciane C Mattos

    Full Text Available BACKGROUND: The unicellular parasite Trypanosoma cruzi is the causative agent of Chagaś disease in humans. Adherence of the infective stage to elements of the extracellular matrix (ECM, as laminin and fibronectin, is an essential step in host cell invasion. Although members of the gp85/TS, as Tc85, were identified as laminin and fibronectin ligands, the signaling events triggered on the parasite upon binding to these molecules are largely unexplored. METHODOLOGY/PRINCIPAL FINDINGS: Viable infective parasites were incubated with laminin, fibronectin or bovine serum albumin for different periods of time and the proteins were separated by bidimensional gels. The phosphoproteins were envisaged by specific staining and the spots showing phosphorylation levels significantly different from the control were excised and identified by MS/MS. The results of interest were confirmed by immunoblotting or immunoprecipitation and the localization of proteins in the parasite was determined by immunofluorescence. Using a host cell-free system, our data indicate that the phosphorylation contents of T. cruzi proteins encompassing different cellular functions are modified upon incubation of the parasite with fibronectin or laminin. CONCLUSIONS/SIGNIFICANCE: Herein it is shown, for the first time, that paraflagellar rod proteins and α-tubulin, major structural elements of the parasite cytoskeleton, are predominantly dephosphorylated during the process, probably involving the ERK1/2 pathway. It is well established that T. cruzi binds to ECM elements during the cell infection process. The fact that laminin and fibronectin induce predominantly dephosphorylation of the main cytoskeletal proteins of the parasite suggests a possible correlation between cytoskeletal modifications and the ability of the parasite to internalize into host cells.

  1. Physical Activity Modifies the Association between Dietary Protein and Lean Mass of Postmenopausal Women.

    Science.gov (United States)

    Martinez, Jessica A; Wertheim, Betsy C; Thomson, Cynthia A; Bea, Jennifer W; Wallace, Robert; Allison, Matthew; Snetselaar, Linda; Chen, Zhao; Nassir, Rami; Thompson, Patricia A

    2017-02-01

    Maintenance of lean muscle mass and related strength is associated with lower risk for numerous chronic diseases of aging in women. Our aim was to evaluate whether the association between dietary protein and lean mass differs by physical activity level, amino acid composition, and body mass index categories. We performed a cross-sectional analysis of a prospective cohort. Participants were postmenopausal women from the Women's Health Initiative with body composition measurements by dual-energy x-ray absorptiometry (n=8,298). Our study measured percent lean mass, percent fat mass, and lean body mass index. Linear regression models adjusted for scanner serial number, age, calibrated energy intake, race/ethnicity, neighborhood socioeconomic status, and recreational physical activity were used to determine the relationship between protein intake and body composition measures. Likelihood ratio tests and stratified analysis were used to investigate physical activity and body mass index as potential effect modifiers. Biomarker-calibrated protein intake was positively associated with percent lean mass; women in the highest protein quintile had 6.3 percentage points higher lean mass than the lowest quintile (Plean body mass index were both inversely related to protein intake (both Plean body mass index (P interaction =0.011). Leucine intake was associated with lean mass, as were branched chain amino acids combined (both Plean mass in postmenopausal women. Importantly, those that also engage in physical activity have the highest lean mass across body mass index categories. Copyright © 2017 Academy of Nutrition and Dietetics. Published by Elsevier Inc. All rights reserved.

  2. Isolation of a macrophage receptor for proteins modified by advanced glycosylation end products

    International Nuclear Information System (INIS)

    Radoff, S.; Vlassara, H.; Cerami, A.

    1987-01-01

    The nonenzymatic reaction of glucose with protein amino groups leads to the formation of irreversible AGE, such as the recently characterized glucose-derived crosslink, [2-furoyl-4(5)-(2-furanyl)-1-H-imidazole] (FFI). These products accumulate with time in aging tissues and diabetes, and are implicated in irreversible tissue damage. The authors have recently shown that macrophages bind and degrade AGE-proteins via a specific surface receptor, which is thus selectively removing senescent macromolecules. Scatchard plot analysis of binding data has indicated 1.5 x 10 5 receptors/cell with a binding affinity (Ka) of 1.7 x 10 7 /M. They have now isolated this receptor from murine macrophage RAW 264.7 membranes, solubilized with octylglucoside/protease inhibitors, and using FFI-Sepharose affinity chromatography and FPLC. The purified receptor binds radioactive FFI-containing compounds competitively. SDS-PAGE gels under reducing conditions indicate the receptor to be composed of two polypeptides, 83 Kda and 36 Kda. Crosslinking experiments with 125 I-AGE-albumin as ligand, indicate the 83 Kda subunit to be the AGE-binding peptide. These studies further characterize a macrophage receptor which selectively recognizes time-dependent glucose-modified proteins associated with aging and diabetes

  3. Spectroscopic detection of fluorescent protein marker gene activity in genetically modified plants

    Science.gov (United States)

    Liew, O. W.; Chong, Jenny P. C.; Asundi, Anand K.

    2005-04-01

    This work focuses on developing a portable fibre optic fluorescence analyser for rapid identification of genetically modified plants tagged with a fluorescent marker gene. Independent transgenic tobacco plant lines expressing the enhanced green fluorescence protein (EGFP) gene were regenerated following Agrobacterium-mediated gene transfer. Molecular characterisation of these plant lines was carried out at the DNA level by PCR screening to confirm their transgenic status. Conventional transgene expression analysis was then carried out at the RNA level by RT-PCR and at the protein level by Western blotting using anti-GFP rabbit antiserum. The amount of plant-expressed EGFP on a Western blot was quantified against known amounts of purified EGFP by scanning densitometry. The expression level of EGFP in transformed plants was found to range from 0.1 - 0.6% of total extractable protein. A comparison between conventional western analysis of transformants and direct spectroscopic quantification using the fibre optic fluorescence analyser was made. The results showed that spectroscopic measurements of fluorescence emission from strong EGFP expressors correlated positively with Western blot data. However, the fluorescence analyser was also able to identify weakly expressing plant transformants below the detection limit of colorimetric Western blotting.

  4. A genetically modified protein-based hydrogel for 3D culture of AD293 cells.

    Directory of Open Access Journals (Sweden)

    Xiao Du

    Full Text Available Hydrogels have strong application prospects for drug delivery, tissue engineering and cell therapy because of their excellent biocompatibility and abundant availability as scaffolds for drugs and cells. In this study, we created hybrid hydrogels based on a genetically modified tax interactive protein-1 (TIP1 by introducing two or four cysteine residues in the primary structure of TIP1. The introduced cysteine residues were crosslinked with a four-armed poly (ethylene glycol having their arm ends capped with maleimide residues (4-armed-PEG-Mal to form hydrogels. In one form of the genetically modification, we incorporated a peptide sequence 'GRGDSP' to introduce bioactivity to the protein, and the resultant hydrogel could provide an excellent environment for a three dimensional cell culture of AD293 cells. The AD293 cells continued to divide and displayed a polyhedron or spindle-shape during the 3-day culture period. Besides, AD293 cells could be easily separated from the cell-gel constructs for future large-scale culture after being cultured for 3 days and treating hydrogel with trypsinase. This work significantly expands the toolbox of recombinant proteins for hydrogel formation, and we believe that our hydrogel will be of considerable interest to those working in cell therapy and controlled drug delivery.

  5. Thermodynamics of protein folding using a modified Wako-Saitô-Muñoz-Eaton model.

    Science.gov (United States)

    Tsai, Min-Yeh; Yuan, Jian-Min; Teranishi, Yoshiaki; Lin, Sheng Hsien

    2012-09-01

    Herein, we propose a modified version of the Wako-Saitô-Muñoz-Eaton (WSME) model. The proposed model introduces an empirical temperature parameter for the hypothetical structural units (i.e., foldons) in proteins to include site-dependent thermodynamic behavior. The thermodynamics for both our proposed model and the original WSME model were investigated. For a system with beta-hairpin topology, a mathematical treatment (contact-pair treatment) to facilitate the calculation of its partition function was developed. The results show that the proposed model provides better insight into the site-dependent thermodynamic behavior of the system, compared with the original WSME model. From this site-dependent point of view, the relationship between probe-dependent experimental results and model's thermodynamic predictions can be explained. The model allows for suggesting a general principle to identify foldon behavior. We also find that the backbone hydrogen bonds may play a role of structural constraints in modulating the cooperative system. Thus, our study may contribute to the understanding of the fundamental principles for the thermodynamics of protein folding.

  6. Mitochondrial associated ubiquitin fold modifier-1 mediated protein conjugation in Leishmania donovani.

    Directory of Open Access Journals (Sweden)

    Sreenivas Gannavaram

    2011-01-01

    Full Text Available In this report, we demonstrate the existence of the ubiquitin fold modifier-1 (Ufm1 and its conjugation pathway in trypanosomatid parasite Leishmania donovani. LdUfm1 is activated by E1-like enzyme LdUba5. LdUfc1 (E2 specifically interacted with LdUfm1 and LdUba5 to conjugate LdUfm1 to proteinaceous targets. Mass spectrometry analysis revealed that LdUfm1 is conjugated to Leishmania protein targets that are associated with mitochondria. Immunofluorescence experiments showed that Leishmania Ufm1, Uba5 and Ufc1 are associated with the mitochondria. The demonstration that all the components of this system as well as the substrates are associated with mitochondrion suggests it may have physiological roles not yet described in any other organism. Overexpression of a non-conjugatable form of LdUfm1 and an active site mutant of LdUba5 resulted in reduced survival of Leishmania in the macrophage. Since mitochondrial activities are developmentally regulated in the life cycle of trypanosomatids, Ufm1 mediated modifications of mitochondrial proteins may be important in such regulation. Thus, Ufm1 conjugation pathway in Leishmania could be explored as a potential drug target in the control of Leishmaniasis.

  7. Pepsin-Assisted Transglutaminase Modifi cation of Functional Properties of a Protein Isolate Obtained from Industrial Sunfl ower Meal

    Directory of Open Access Journals (Sweden)

    Petya Ivanova

    2017-01-01

    Full Text Available The utilization of industrial sunfl ower meal to produce protein-rich products for the food industry is an alternative approach for bett er and more effi cient use of this agricultural by-product. Sunfl ower meal proteins possess specifi c functional properties, which however need improvement to broaden their potential as supplements for delivering high-quality products for human nutrition. The aim of the study is to evaluate the combined infl uence of low-degree pepsin hydrolysis and transglutaminase (TG modifi cation on industrial sunfl ower meal protein isolate functionality at pH=2 to 10. Three TG-modifi ed pepsin hydrolysates with the degree of hydrolysis of 0.48, 0.71 and 1.72 % were produced and named TG-PH1, TG-PH2 and TG-PH3, respectively. All three TG-modifi ed pepsin hydrolysates exhibited improved solubility at pH between 3.5 and 5.5 as the highest was observed of TG-PH3 at protein isoelectric point (pI=4.5. Sunfl ower meal protein isolate and TG-modifi ed sunfl ower meal protein isolate had greater solubility than the three TG-modifi ed hydrolysates at pH7. Signifi cant improvement of foam making capacity (p<0.05 was achieved with all three TG-modifi ed pepsin hydrolysates in the entire pH area studied. Pepsin hydrolysis of the protein isolate with the three degrees of hydrolysis did not improve foam stability. Improved thermal stability was observed with TG-PH3 up to 80 °C compared to the protein isolate (pH=7. At 90 °C, TG modifi cation of the protein isolate alone resulted in the highest thermal stability. Pepsin hydrolysis followed by a treatment with TG could be used to produce sunfl ower protein isolates with improved solubility, foam making capacity and thermal stability for use in the food industry.

  8. Development of Cy5.5-Labeled Hydrophobically Modified Glycol Chitosan Nanoparticles for Protein Delivery

    Science.gov (United States)

    Chin, Amanda

    Therapeutic proteins are often highly susceptible to enzymatic degradation, thus restricting their in vivo stability. To overcome this limitation, delivery systems designed to promote uptake and reduce degradation kinetics have undergone a rapid shift from macro-scale systems to nanomaterial based carriers. Many of these nanomaterials, however, elicit immune responses and may have cytotoxic effects both in vitro and in vivo. The naturally derived polysaccharide chitosan has emerged as a promising biodegradable material and has been utilized for many biomedical applications; nevertheless, its function is often constrained by poor solubility. Glycol chitosan, a derivative of chitosan, can be hydrophobically modified to impart amphiphilic properties that enable the self-assembly into nanoparticles in aqueous media at neutral pH. This nanoparticle system has shown initial success as a therapeutic agent in several model cell culture systems, but little is known about its stability against enzymatic degradation. Therefore, the goal of this research was to investigate the resistance of hydrophobically modified glycol chitosan against enzyme-catalyzed degradation using an in vivo simulated system containing lysozyme. To synthesize the nanoparticles, hydrophobic cholanic acid was first covalently conjugated to glycol chitosan using of N-(3-Dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS). Conjugates were purified by dialysis, lyophilized, and ultra-sonicated to form nanoparticles. Fourier transform infrared (FT-IR) spectroscopy confirmed the binding of 5beta-cholanic acid to the glycol chitosan. Particle size and stability over time were determined with dynamic light scattering (DLS), and particle morphology was evaluated by transmission electron microscopy (TEM). The average diameter of the nanoparticles was approximately 200 nm, which remained stable at 4°C for up to 10 days. Additionally, a near infrared fluorescent (NIRF) dye

  9. Median Modified Wiener Filter for nonlinear adaptive spatial denoising of protein NMR multidimensional spectra

    KAUST Repository

    Cannistraci, Carlo Vittorio

    2015-01-26

    Denoising multidimensional NMR-spectra is a fundamental step in NMR protein structure determination. The state-of-the-art method uses wavelet-denoising, which may suffer when applied to non-stationary signals affected by Gaussian-white-noise mixed with strong impulsive artifacts, like those in multi-dimensional NMR-spectra. Regrettably, Wavelet\\'s performance depends on a combinatorial search of wavelet shapes and parameters; and multi-dimensional extension of wavelet-denoising is highly non-trivial, which hampers its application to multidimensional NMR-spectra. Here, we endorse a diverse philosophy of denoising NMR-spectra: less is more! We consider spatial filters that have only one parameter to tune: the window-size. We propose, for the first time, the 3D extension of the median-modified-Wiener-filter (MMWF), an adaptive variant of the median-filter, and also its novel variation named MMWF*. We test the proposed filters and the Wiener-filter, an adaptive variant of the mean-filter, on a benchmark set that contains 16 two-dimensional and three-dimensional NMR-spectra extracted from eight proteins. Our results demonstrate that the adaptive spatial filters significantly outperform their non-adaptive versions. The performance of the new MMWF* on 2D/3D-spectra is even better than wavelet-denoising. Noticeably, MMWF* produces stable high performance almost invariant for diverse window-size settings: this signifies a consistent advantage in the implementation of automatic pipelines for protein NMR-spectra analysis.

  10. Median Modified Wiener Filter for nonlinear adaptive spatial denoising of protein NMR multidimensional spectra

    KAUST Repository

    Cannistraci, Carlo Vittorio; Abbas, Ahmed; Gao, Xin

    2015-01-01

    Denoising multidimensional NMR-spectra is a fundamental step in NMR protein structure determination. The state-of-the-art method uses wavelet-denoising, which may suffer when applied to non-stationary signals affected by Gaussian-white-noise mixed with strong impulsive artifacts, like those in multi-dimensional NMR-spectra. Regrettably, Wavelet's performance depends on a combinatorial search of wavelet shapes and parameters; and multi-dimensional extension of wavelet-denoising is highly non-trivial, which hampers its application to multidimensional NMR-spectra. Here, we endorse a diverse philosophy of denoising NMR-spectra: less is more! We consider spatial filters that have only one parameter to tune: the window-size. We propose, for the first time, the 3D extension of the median-modified-Wiener-filter (MMWF), an adaptive variant of the median-filter, and also its novel variation named MMWF*. We test the proposed filters and the Wiener-filter, an adaptive variant of the mean-filter, on a benchmark set that contains 16 two-dimensional and three-dimensional NMR-spectra extracted from eight proteins. Our results demonstrate that the adaptive spatial filters significantly outperform their non-adaptive versions. The performance of the new MMWF* on 2D/3D-spectra is even better than wavelet-denoising. Noticeably, MMWF* produces stable high performance almost invariant for diverse window-size settings: this signifies a consistent advantage in the implementation of automatic pipelines for protein NMR-spectra analysis.

  11. Protein adsorption capability on polyurethane and modified-polyurethane membrane for periodontal guided tissue regeneration applications

    Energy Technology Data Exchange (ETDEWEB)

    Sheikh, Zeeshan [Matrix Dynamics Group, Faculty of Dentistry, University of Toronto, Fitzgerald Building, 150 College Street, Toronto, ON M5S 3E2 (Canada); School of Engineering and Materials Science, Queen Mary, University of London, Mile End Rd, London, E1 4NS (United Kingdom); Khan, Abdul Samad, E-mail: draskhan@ciitlahore.edu.pk [Interdisciplinary Research Centre in Biomedical Materials, COMSATS Institute of Information Technology, Lahore 54000 (Pakistan); Roohpour, Nima [Oral Care R& D, GSK St., Georges Ave., Weybridge KT13 8PA (United Kingdom); Glogauer, Michael [Matrix Dynamics Group, Faculty of Dentistry, University of Toronto, Fitzgerald Building, 150 College Street, Toronto, ON M5S 3E2 (Canada); Rehman, Ihtesham u [Department of Materials Science and Engineering, The Kroto Research Institute, North Campus, University of Sheffield, Broad Lane, Sheffield S3 7HQ (United Kingdom)

    2016-11-01

    Periodontal disease if left untreated can result in creation of defects within the alveolar ridge. Barrier membranes are frequently used with or without bone replacement graft materials for achieving periodontal guided tissue regeneration (GTR). Surface properties of barrier membranes play a vital role in their functionality and clinical success. In this study polyetherurethane (PEU) membranes were synthesized by using 4,4′-methylene-diphenyl diisocyanate (MDI), polytetramethylene oxide (PTMO) and 1,4-butane diol (BDO) as a chain extender via solution polymerization. Hydroxyl terminated polydimethylsiloxane (PDMS) due to having inherent surface orientation towards air was used for surface modification of PEU on one side of the membranes. This resulting membranes had one surface being PEU and the other being PDMS coated PEU. The prepared membranes were treated with solutions of bovine serum albumin (BSA) in de-ionized water at 37 °C at a pH of 7.2. The surface protein adsorptive potential of PEU membranes was observed using Attenuated Total Reflectance Fourier Transform Infrared Spectroscopy (ATR-FTIR), Raman spectroscopy and Confocal Raman spectroscopy. The contact angle measurement, tensile strength and modulus of prepared membranes were also evaluated. PEU membrane (89.86 ± 1.62°) exhibited less hydrophobic behavior than PEU-PDMS (105.87 ± 3.16°). The ultimate tensile strength and elastic modulus of PEU (27 ± 1 MPa and 14 ± 2 MPa) and PEU-PDMS (8 ± 1 MPa and 26 ± 1 MPa) membranes was in required range. The spectral analysis revealed adsorption of BSA proteins on the surface of non PDMS coated PEU surface. The PDMS modified PEU membranes demonstrated a lack of BSA adsorption. The non PDMS coated side of the membrane which adsorbs proteins could potentially be used facing towards the defect attracting growth factors for periodontal tissue regeneration. Whereas, the PDMS coated side could serve as an occlusive barrier for preventing gingival epithelial

  12. Protein adsorption capability on polyurethane and modified-polyurethane membrane for periodontal guided tissue regeneration applications

    International Nuclear Information System (INIS)

    Sheikh, Zeeshan; Khan, Abdul Samad; Roohpour, Nima; Glogauer, Michael; Rehman, Ihtesham u

    2016-01-01

    Periodontal disease if left untreated can result in creation of defects within the alveolar ridge. Barrier membranes are frequently used with or without bone replacement graft materials for achieving periodontal guided tissue regeneration (GTR). Surface properties of barrier membranes play a vital role in their functionality and clinical success. In this study polyetherurethane (PEU) membranes were synthesized by using 4,4′-methylene-diphenyl diisocyanate (MDI), polytetramethylene oxide (PTMO) and 1,4-butane diol (BDO) as a chain extender via solution polymerization. Hydroxyl terminated polydimethylsiloxane (PDMS) due to having inherent surface orientation towards air was used for surface modification of PEU on one side of the membranes. This resulting membranes had one surface being PEU and the other being PDMS coated PEU. The prepared membranes were treated with solutions of bovine serum albumin (BSA) in de-ionized water at 37 °C at a pH of 7.2. The surface protein adsorptive potential of PEU membranes was observed using Attenuated Total Reflectance Fourier Transform Infrared Spectroscopy (ATR-FTIR), Raman spectroscopy and Confocal Raman spectroscopy. The contact angle measurement, tensile strength and modulus of prepared membranes were also evaluated. PEU membrane (89.86 ± 1.62°) exhibited less hydrophobic behavior than PEU-PDMS (105.87 ± 3.16°). The ultimate tensile strength and elastic modulus of PEU (27 ± 1 MPa and 14 ± 2 MPa) and PEU-PDMS (8 ± 1 MPa and 26 ± 1 MPa) membranes was in required range. The spectral analysis revealed adsorption of BSA proteins on the surface of non PDMS coated PEU surface. The PDMS modified PEU membranes demonstrated a lack of BSA adsorption. The non PDMS coated side of the membrane which adsorbs proteins could potentially be used facing towards the defect attracting growth factors for periodontal tissue regeneration. Whereas, the PDMS coated side could serve as an occlusive barrier for preventing gingival epithelial

  13. Radiochemical verification and validation in the environmental data collection process

    International Nuclear Information System (INIS)

    Rosano-Reece, D.; Bottrell, D.; Bath, R.J.

    1994-01-01

    A credible and cost effective environmental data collection process should produce analytical data which meets regulatory and program specific requirements. Analytical data, which support the sampling and analysis activities at hazardous waste sites, undergo verification and independent validation before the data are submitted to regulators. Understanding the difference between verification and validation and their respective roles in the sampling and analysis process is critical to the effectiveness of a program. Verification is deciding whether the measurement data obtained are what was requested. The verification process determines whether all the requirements were met. Validation is more complicated than verification. It attempts to assess the impacts on data use, especially when requirements are not met. Validation becomes part of the decision-making process. Radiochemical data consists of a sample result with an associated error. Therefore, radiochemical validation is different and more quantitative than is currently possible for the validation of hazardous chemical data. Radiochemical data include both results and uncertainty that can be statistically compared to identify significance of differences in a more technically defensible manner. Radiochemical validation makes decisions about analyte identification, detection, and uncertainty for a batch of data. The process focuses on the variability of the data in the context of the decision to be made. The objectives of this paper are to present radiochemical verification and validation for environmental data and to distinguish the differences between the two operations

  14. Screening for Glycosylphosphatidylinositol-Modified Cell Wall Proteins in Pichia pastoris and Their Recombinant Expression on the Cell Surface

    Science.gov (United States)

    Zhang, Li; Liang, Shuli; Zhou, Xinying; Jin, Zi; Jiang, Fengchun; Han, Shuangyan; Zheng, Suiping

    2013-01-01

    Glycosylphosphatidylinositol (GPI)-anchored glycoproteins have various intrinsic functions in yeasts and different uses in vitro. In the present study, the genome of Pichia pastoris GS115 was screened for potential GPI-modified cell wall proteins. Fifty putative GPI-anchored proteins were selected on the basis of (i) the presence of a C-terminal GPI attachment signal sequence, (ii) the presence of an N-terminal signal sequence for secretion, and (iii) the absence of transmembrane domains in mature protein. The predicted GPI-anchored proteins were fused to an alpha-factor secretion signal as a substitute for their own N-terminal signal peptides and tagged with the chimeric reporters FLAG tag and mature Candida antarctica lipase B (CALB). The expression of fusion proteins on the cell surface of P. pastoris GS115 was determined by whole-cell flow cytometry and immunoblotting analysis of the cell wall extracts obtained by β-1,3-glucanase digestion. CALB displayed on the cell surface of P. pastoris GS115 with the predicted GPI-anchored proteins was examined on the basis of potential hydrolysis of p-nitrophenyl butyrate. Finally, 13 proteins were confirmed to be GPI-modified cell wall proteins in P. pastoris GS115, which can be used to display heterologous proteins on the yeast cell surface. PMID:23835174

  15. Quantitative analysis of modified proteins and their positional isomers by tandem mass spectrometry: human histone H4.

    Science.gov (United States)

    Pesavento, James J; Mizzen, Craig A; Kelleher, Neil L

    2006-07-01

    Here we show that fragment ion abundances from dissociation of ions created from mixtures of multiply modified histone H4 (11 kDa) or of N-terminal synthetic peptides (2 kDa) correspond to their respective intact ion abundances measured by Fourier transform mass spectrometry. Isomeric mixtures of modified forms of the same protein are resolved and quantitated with a precision of protein ions created by electrospray greatly easing many of the systematic biases that more strongly affect small peptides (e.g., differences in ionization efficiency and ion m/z values). The ion fragmentation methods validated here are directly extensible to intact human proteins to derive quantitative information on the highly related and often isomeric protein forms created by combinatorial arrays of posttranslational modifications.

  16. Functionality of whey proteins covalently modified by allyl isothiocyanate. Part 1 physicochemical and antibacterial properties of native and modified whey proteins at pH 2 to 7

    NARCIS (Netherlands)

    Keppler, Julia Katharina; Martin, Dierk; Garamus, Vasil M.; Berton-Carabin, Claire; Nipoti, Elia; Coenye, Tom; Schwarz, Karin

    2017-01-01

    Whey protein isolate (WPI) (∼75% β-lactoglobulin (β-LG)) is frequently used in foods as a natural emulsifying agent. However, at an acidic pH value, its emulsification capacity is greatly reduced. The covalent attachment of natural electrophilic hydrophobic molecules to WPI proteins is a

  17. Phenolsulphotransferase in human tissue: radiochemical enzymatic assay and biochemical properties

    International Nuclear Information System (INIS)

    Anderson, R.J.; Weinshilboum, R.M.

    1980-01-01

    Phenolsulphotransferase (EC 2.8.2.1) (PST) is an important catecholamine and drug metabolizing enzyme. Optimal conditions have been determined for the accurate measurement of PST activity in the human platelet, human renal cortex, and human jejunum with a radiochemical microassay. 3-Methoxy-4-hydroxyphenylglycol (MHPG) and 35 S-3'-phosphoadenosine-5'-phosphosulfate ( 35 S-PAPS) were the substrates for the reaction. The apparent Michaelis-Menten (Ksub(m)) values for MHPG with platelet, renal cortex, and jejunum were 1.09, 0.46 and 1.16 mmol/l, respectively. Apparent Ksub(m) values for PAPS in the same tissues were 0.14, 0.13 and 0.21 μmol/l. The pH optimum of the reacton in all three tissues was approximately 6.2-6.8 with three different buffer systems. The coefficients of variation for the assay of platelet, renal cortex, and jejunal activities were 6.2%, 3.4% and 4.4%, respectively. Mean platelet PST activity in blood samples from 75 randomly selected adult subjects was 5.0 +- 1.72 mmol of MHPG sulfate formed per hour per mg of platelet protein (8.3 X 10 -5 +- 2.9 X 10 -5 μmol min -1 mg -1 , mean +- S.D.). There was a 5-fold intersubject variation in platelet PST activity within two standard deviations of the mean value. Experiments in which partially purified human erythrocyte PST was added to platelet, kidney and gut homogenates under these assay conditions provided evidence that endogenous PST inhibitors did not affect the observed enzyme activity. (Auth.)

  18. Radiochemical studies on environmental radioactivity in Sudan

    Energy Technology Data Exchange (ETDEWEB)

    Sam, Adam Khatir [Sudan Atomic Energy Commission, Khartoum (Sudan)

    1998-09-01

    Measurements of uranium and thorium isotopes, {sup 226} Ra, {sup 210} Po, {sup 228} Ra, {sup 40} K and fallout radionuclide {sup 137} Cs in soil samples collected from different districts in Sudan, rock phosphate samples collected from the uro and kurun rock phosphate deposits in the eastern part of the Nuba mountains in Western Sudan, and surface marine sediments and marine organisms collected from the sudanese coastal waters of the Red Sea have been made using a high resolution gamma-spectrometry, radiochemical separation and {alpha} spectrometry. The external exposure due to {gamma} radiation from the ground has been calculated. The average exposure was found to be 45.4 {+-} 21.3 nGy/h, corresponding to the annual dose equivalent of 278 {mu}Sv/y. With the exception of some areas, the calculated exposure falls within the global wide range of outdoor radiation exposure given in the UNSCEAR publications. The nation-wide average concentrations of {sup 226} Ra, {sup 238} U, {sup 232} Th, {sup 40} K and {sup 137} Cs determined were 31.6 {+-} 27, 20.1 {+-} 16.4, 19.1 {+-} 8.1, 280.3 {+-} 137.6 and 4.1 {+-} 4.3 Bq/Kg, respectively. This shows that there is little contamination due to fallout radioactivity at survey sites. The exchangeable radium fraction constitutes 19-24% of the total radium content. The data show that {sup 238} U and its decay products are the principal contributors of radioactivity in both phosphate deposits at Uro and Kurun. The equivalent mass concentrations of uranium in the Uro rock phosphate fall within the range that could be economically recovered as the by-product of fertilizer industry. The mean activity concentrations weighted by average agricultural consumption of 300 kg/ha of untreated ground rock fertilizer resulted in an annual distribution of 120.63 Bq Ra/m{sup 2} with Uro rock and 12.97, 0.21 and 4.24 Bq/m{sup 2} respectively, with Kurun rock fertilizer. The external radiation exposure over agricultural areas was estimated 23.41 x 10

  19. Radiochemical studies on environmental radioactivity in Sudan

    International Nuclear Information System (INIS)

    Sam, Adam Khatir

    1998-01-01

    Measurements of uranium and thorium isotopes, 226 Ra, 210 Po, 228 Ra, 40 K and fallout radionuclide 137 Cs in soil samples collected from different districts in Sudan, rock phosphate samples collected from the uro and kurun rock phosphate deposits in the eastern part of the Nuba mountains in Western Sudan, and surface marine sediments and marine organisms collected from the sudanese coastal waters of the Red Sea have been made using a high resolution gamma-spectrometry, radiochemical separation and α spectrometry. The external exposure due to γ radiation from the ground has been calculated. The average exposure was found to be 45.4 ± 21.3 nGy/h, corresponding to the annual dose equivalent of 278 μSv/y. With the exception of some areas, the calculated exposure falls within the global wide range of outdoor radiation exposure given in the UNSCEAR publications. The nation-wide average concentrations of 226 Ra, 238 U, 232 Th, 40 K and 137 Cs determined were 31.6 ± 27, 20.1 ± 16.4, 19.1 ± 8.1, 280.3 ± 137.6 and 4.1 ± 4.3 Bq/Kg, respectively. This shows that there is little contamination due to fallout radioactivity at survey sites. The exchangeable radium fraction constitutes 19-24% of the total radium content. The data show that 238 U and its decay products are the principal contributors of radioactivity in both phosphate deposits at Uro and Kurun. The equivalent mass concentrations of uranium in the Uro rock phosphate fall within the range that could be economically recovered as the by-product of fertilizer industry. The mean activity concentrations weighted by average agricultural consumption of 300 kg/ha of untreated ground rock fertilizer resulted in an annual distribution of 120.63 Bq Ra/m 2 with Uro rock and 12.97, 0.21 and 4.24 Bq/m 2 respectively, with Kurun rock fertilizer. The external radiation exposure over agricultural areas was estimated 23.41 x 10 -9 Gy/h and 2.59 x 10 -9 Gy/h at 1 m above ground level for Uro and Kurun rock phosphate fertilizers

  20. Radiochemical studies of some preparation methods for phosphorus

    International Nuclear Information System (INIS)

    Loos-Neskovic, C.; Fedoroff, M.

    1983-01-01

    Various methods of radiochemical separation were tested for the determination of phosphorus in metals and alloys by neutron activation analysis. Classical methods of separation revealed some defects when they were applied to this problem. Methods using liquid extraction gave low yields and were not reproducible. Methods based on precipitation gave better results, but were not selective enough in most cases. Retention on alumina was not possible without preliminary separations. Authors studied a new radiochemical separation based on the extraction of elemental phosphorus in the gaseous phase after reduction at high temperature with carbon. Measurements with radioactive phosphorus showed that the extraction yield is better than 99%. (author)

  1. Radiochemical analysis for nuclear waste management in decommissioning

    International Nuclear Information System (INIS)

    Hou, X.

    2010-07-01

    The NKS-B RadWaste project was launched from June 2009. The on-going decommissioning activities in Nordic countries and current requirements and problems on the radiochemical analysis of decommissioning waste were discussed and overviewed. The radiochemical analytical methods used for determination of various radionuclides in nuclear waste are reviewed, a book was written by the project partners Jukka Lehto and Xiaolin Hou on the chemistry and analysis of radionuclide to be published in 2010. A summary of the methods developed in Nordic laboratories is described in this report. The progresses on the development and optimization of analytical method in the Nordic labs under this project are presented. (author)

  2. Radiochemical analysis for nuclear waste management in decommissioning

    Energy Technology Data Exchange (ETDEWEB)

    Hou, X. (Technical Univ. of Denmark, Risoe National Lab. for Sustainable Energy. Radiation Research Div., Roskilde (Denmark))

    2010-07-15

    The NKS-B RadWaste project was launched from June 2009. The on-going decommissioning activities in Nordic countries and current requirements and problems on the radiochemical analysis of decommissioning waste were discussed and overviewed. The radiochemical analytical methods used for determination of various radionuclides in nuclear waste are reviewed, a book was written by the project partners Jukka Lehto and Xiaolin Hou on the chemistry and analysis of radionuclide to be published in 2010. A summary of the methods developed in Nordic laboratories is described in this report. The progresses on the development and optimization of analytical method in the Nordic labs under this project are presented. (author)

  3. Translationally controlled tumor protein supplemented chitosan modified glass ionomer cement promotes osteoblast proliferation and function

    International Nuclear Information System (INIS)

    Sangsuwan, Jiraporn; Wanichpakorn, Supreya; Kedjarune-Leggat, Ureporn

    2015-01-01

    The objective of this study was to evaluate the effect of translationally controlled tumor protein (TCTP) supplemented in a novel glass ionomer cement (BIO-GIC) on normal human osteoblasts (NHost cells). BIO-GIC was a glass ionomer cement (GIC) modified by adding chitosan and albumin to promote the release of TCTP. NHost cells were seeded on specimens of GIC, GIC + TCTP, BIO-GIC and BIO-GIC + TCTP. Cell proliferation was determined by BrdU assay. It was found that BIO-GIC + TCTP had significantly higher proliferation of cells than other specimens. Bone morphogenetic protein-2 (BMP-2) and osteopontin (OPN) gene expressions assessed by quantitative real time PCR and alkaline phosphatase (ALP) activity were used to determine cell differentiation. Bone cell function was investigated by calcium deposition using alizarin assay. Both BMP-2 and OPN gene expressions of cells cultured on specimens with added TCTP increased gradually up-regulation after day 1 and reached the highest on day 3 then down-regulation on day 7. The ALP activity of cells cultured on BIO-GIC + TCTP for 7 days and calcium content after 14 days were significantly higher than other groups. BIO-GIC + TCTP can promote osteoblast cells proliferation, differentiation and function. - Highlights: • Developed a new GIC by supplementing TCTP in BIO-GIC (GIC with chitosan and albumin) • BIO-GIC + TCTP released a higher amount of TCTP than GIC + TCTP. • BIO-GIC + TCTP promoted cell proliferation higher than other specimens and control. • BIO-GIC + TCTP promoted osteoblasts differentiation and function

  4. Translationally controlled tumor protein supplemented chitosan modified glass ionomer cement promotes osteoblast proliferation and function

    Energy Technology Data Exchange (ETDEWEB)

    Sangsuwan, Jiraporn [Department of Molecular Biology and Bioinformatics, Center for Genomics and Bioinformatics Research, Faculty of Science, Prince of Songkla University, Hat Yai, Songkhla 90112 (Thailand); Department of Oral Biology and Occlusion, Faculty of Dentistry, Prince of Songkla University, Hat Yai, Songkhla 90112 (Thailand); Wanichpakorn, Supreya; Kedjarune-Leggat, Ureporn [Department of Oral Biology and Occlusion, Faculty of Dentistry, Prince of Songkla University, Hat Yai, Songkhla 90112 (Thailand)

    2015-09-01

    The objective of this study was to evaluate the effect of translationally controlled tumor protein (TCTP) supplemented in a novel glass ionomer cement (BIO-GIC) on normal human osteoblasts (NHost cells). BIO-GIC was a glass ionomer cement (GIC) modified by adding chitosan and albumin to promote the release of TCTP. NHost cells were seeded on specimens of GIC, GIC + TCTP, BIO-GIC and BIO-GIC + TCTP. Cell proliferation was determined by BrdU assay. It was found that BIO-GIC + TCTP had significantly higher proliferation of cells than other specimens. Bone morphogenetic protein-2 (BMP-2) and osteopontin (OPN) gene expressions assessed by quantitative real time PCR and alkaline phosphatase (ALP) activity were used to determine cell differentiation. Bone cell function was investigated by calcium deposition using alizarin assay. Both BMP-2 and OPN gene expressions of cells cultured on specimens with added TCTP increased gradually up-regulation after day 1 and reached the highest on day 3 then down-regulation on day 7. The ALP activity of cells cultured on BIO-GIC + TCTP for 7 days and calcium content after 14 days were significantly higher than other groups. BIO-GIC + TCTP can promote osteoblast cells proliferation, differentiation and function. - Highlights: • Developed a new GIC by supplementing TCTP in BIO-GIC (GIC with chitosan and albumin) • BIO-GIC + TCTP released a higher amount of TCTP than GIC + TCTP. • BIO-GIC + TCTP promoted cell proliferation higher than other specimens and control. • BIO-GIC + TCTP promoted osteoblasts differentiation and function.

  5. Middle east respiratory syndrome coronavirus spike protein delivered by modified vaccinia virus ankara efficiently induces virus-neutralizing antibodies

    NARCIS (Netherlands)

    F. Song (Fei); R. Fux (Robert); L.B.V. Provacia (Lisette); A. Volz (Asisa); M. Eickmann; S. Becker (Stephan); A.D.M.E. Osterhaus (Albert); B.L. Haagmans (Bart); G. Suttera (Gerd)

    2013-01-01

    textabstractMiddle East respiratory syndrome coronavirus (MERS-CoV) has recently emerged as a causative agent of severe respiratory disease in humans. Here, we constructed recombinant modified vaccinia virus Ankara (MVA) expressing full-length MERS-CoV spike (S) protein (MVA-MERS-S). The genetic

  6. Preparation of C-terminally modified chemokines by expressed protein ligation.

    Science.gov (United States)

    Baumann, Lars; Steinhagen, Max; Beck-Sickinger, Annette G

    2013-01-01

    In order to link structural features on a molecular level to the function of chemokines, site-specific modification strategies are strongly required. These can be used to incorporate fluorescent dyes and/or physical probes to allow investigations in a wide range of biological and physical techniques, e.g., nuclear magnetic resonance (NMR) spectroscopy, fluorescence microscopy, fluorescence resonance energy transfer (FRET), or fluorescence correlation spectroscopy (FCS). Only a limited number of functional groups within the 20 canonical amino acids allow ligation strategies that can be helpful to introduce novel functionalities, which in turn expand the scope of chemoselective and orthogonal reactivity of (semi)synthetic chemokines. In the present chapter we mainly focus on the fabulous history of native chemical ligation (NCL) and provide a general protocol for the preparation of C-terminally modified SDF-1α including tips and tricks for practical work. We believe that this protocol can be easily adapted to other chemokines and many proteins in general.

  7. Branched-chain Amino Acid Biosensing Using Fluorescent Modified Engineered Leucine/Isoleucine/Valine Binding Protein

    Directory of Open Access Journals (Sweden)

    Koji Sode

    2007-06-01

    Full Text Available A novel fluorescence sensing system for branched-chain amino acids (BCAAswas developed based on engineered leucine/isoleucine/valine-binding proteins (LIVBPsconjugated with environmentally sensitive fluorescence probes. LIVBP was cloned fromEscherichia coli and Gln149Cys, Gly227Cys, and Gln254Cys mutants were generated bygenetic engineering. The mutant LIVBPs were then modified with environmentallysensitive fluorophores. Based on the fluorescence intensity change observed upon thebinding of the ligands, the MIANS-conjugated Gln149Cys mutant (Gln149Cys-M showedthe highest and most sensitive response. The BCAAs Leu, Ile, and Val can each bemonitored at the sub-micromolar level using Gln149Cys-M. Measurements were alsocarried out on a mixture of BCAFAs and revealed that Gln149Cys-M-based measurementis not significantly affected by the change in the molar ratio of Leu, Ile and Val in thesample. Its high sensitivity and group-specific molecular recognition ability make the newsensing system ideally suited for the measurement of BCAAs and the determination of theFischer ratio, an indicator of hepatic disease involving metabolic dysfunction.

  8. Modifying the properties of whey protein isolate edible film by incorporating palm oil and glycerol

    Directory of Open Access Journals (Sweden)

    Vachiraya Liaotrakoon

    2018-02-01

    Full Text Available This study aimed to improve the properties of whey protein isolate (WPI films by incorporating palm oil (6, 7, and 8% w/w and glycerol (40, 50 and 60% w/w. The lightness of the films increased as glycerol levels increased, but the redness increased with the increased amount of oil content. Increasing the amounts of palm oil and glycerol improved flexibility (P<0.05, but reduced the strength of the film (P<0.05. Films with higher levels of palm oil and lower amounts of glycerol were less permeable to water vapor and oxygen, but more thermally stable. The size of particles and air bubbles in the films reduced with increased palm oil content, regardless of glycerol level. Among all formulae, the film prepared with 8% palm oil and 40% glycerol showed the best overall results. Modifying WPI films with palm oil and glycerol offers a simple technique for producing packaging with better environmental barrier properties.

  9. Improved mucoadhesion and cell uptake of chitosan and chitosan oligosaccharide surface-modified polymer nanoparticles for mucosal delivery of proteins.

    Science.gov (United States)

    Dyawanapelly, Sathish; Koli, Uday; Dharamdasani, Vimisha; Jain, Ratnesh; Dandekar, Prajakta

    2016-08-01

    The main aim of the present study was to compare mucoadhesion and cellular uptake efficiency of chitosan (CS) and chitosan oligosaccharide (COS) surface-modified polymer nanoparticles (NPs) for mucosal delivery of proteins. We have developed poly (D, L-lactide-co-glycolide) (PLGA) NPs, surface-modified COS-PLGA NPs and CS-PLGA NPs, by using double emulsion solvent evaporation method, for encapsulating bovine serum albumin (BSA) as a model protein. Surface modification of NPs was confirmed using physicochemical characterization methods such as particle size and zeta potential, SEM, TEM and FTIR analysis. Both surface-modified PLGA NPs displayed a slow release of protein compared to PLGA NPs. Furthermore, we have explored the mucoadhesive property of COS as a material for modifying the surface of polymeric NPs. During in vitro mucoadhesion test, positively charged COS-PLGA NPs and CS-PLGA NPs exhibited enhanced mucoadhesion, compared to negatively charged PLGA NPs. This interaction was anticipated to improve the cell interaction and uptake of NPs, which is an important requirement for mucosal delivery of proteins. All nanoformulations were found to be safe for cellular delivery when evaluated in A549 cells. Moreover, intracellular uptake behaviour of FITC-BSA loaded NPs was extensively investigated by confocal laser scanning microscopy and flow cytometry. As we hypothesized, positively charged COS-PLGA NPs and CS-PLGA NPs displayed enhanced intracellular uptake compared to negatively charged PLGA NPs. Our results demonstrated that CS- and COS-modified polymer NPs could be promising carriers for proteins, drugs and nucleic acids via nasal, oral, buccal, ocular and vaginal mucosal routes.

  10. Physical Stability of Octenyl Succinate-Modified Polysaccharides and Whey Proteins for Potential Use as Bioactive Carriers in Food Systems.

    Science.gov (United States)

    Puerta-Gomez, Alex F; Castell-Perez, M Elena

    2015-06-01

    The high cost and potential toxicity of biodegradable polymers like poly(lactic-co-glycolic)acid (PLGA) has increased the interest in natural and modified biopolymers as bioactive carriers. This study characterized the physical stability (water sorption and state transition behavior) of selected starch and proteins: octenyl succinate-modified depolymerized waxy corn starch (DWxCn), waxy rice starch (DWxRc), phytoglycogen, whey protein concentrate (80%, WPC), whey protein isolate (WPI), and α-lactalbumin (α-L) to determine their potential as carriers of bioactive compounds under different environmental conditions. After enzyme modification and particle size characterization, glass transition temperature and moisture isotherms were used to characterize the systems. DWxCn and DWxRc had increased water sorption compared to native starch. The level of octenyl succinate anhydrate (OSA) modification (3% and 7%) did not reduce the water sorption of the DWxCn and phytoglycogen samples. The Guggenheim-Andersen-de Boer model indicated that native waxy corn had significantly (P whey proteins had higher glass transition temperature (Tg) values. On the other hand, depolymerized waxy starches at 7%-OSA modification had a "melted" appearance when exposed to environments with high relative humidity (above 70%) after 10 days at 23 °C. The use of depolymerized and OSA-modified polysaccharides blended with proteins created more stable blends of biopolymers. Hence, this biopolymer would be suitable for materials exposed to high humidity environments in food applications. © 2015 Institute of Food Technologists®

  11. Optimized Mitochondrial Targeting of Proteins Encoded by Modified mRNAs Rescues Cells Harboring Mutations in mtATP6

    Directory of Open Access Journals (Sweden)

    Randall Marcelo Chin

    2018-03-01

    Full Text Available Summary: Mitochondrial disease may be caused by mutations in the protein-coding genes of the mitochondrial genome. A promising strategy for treating such diseases is allotopic expression—the translation of wild-type copies of these proteins in the cytosol, with subsequent translocation into the mitochondria, resulting in rescue of mitochondrial function. In this paper, we develop an automated, quantitative, and unbiased screening platform to evaluate protein localization and mitochondrial morphology. This platform was used to compare 31 mitochondrial targeting sequences and 15 3′ UTRs in their ability to localize up to 9 allotopically expressed proteins to the mitochondria and their subsequent impact on mitochondrial morphology. Taking these two factors together, we synthesized chemically modified mRNAs that encode for an optimized allotopic expression construct for mtATP6. These mRNAs were able to functionally rescue a cell line harboring the 8993T > G point mutation in the mtATP6 gene. : Allotopic expression of proteins normally encoded by mtDNA is a promising therapy for mitochondrial disease. Chin et al. use an unbiased and high-content imaging-based screening platform to optimize allotopic expression. Modified mRNAs encoding for the optimized allotopic expression constructs rescued the respiration and growth of mtATP6-deficient cells. Keywords: mitochondria, mitochondrial disease, mRNA, modified mRNA, ATP6, allotopic expression, rare disease, gene therapy, screening, high content imaging

  12. Integration of carboxyl modified magnetic particles and aqueous two-phase extraction for selective separation of proteins.

    Science.gov (United States)

    Gai, Qingqing; Qu, Feng; Zhang, Tao; Zhang, Yukui

    2011-07-15

    Both of the magnetic particle adsorption and aqueous two-phase extraction (ATPE) were simple, fast and low-cost method for protein separation. Selective proteins adsorption by carboxyl modified magnetic particles was investigated according to protein isoelectric point, solution pH and ionic strength. Aqueous two-phase system of PEG/sulphate exhibited selective separation and extraction for proteins before and after magnetic adsorption. The two combination ways, magnetic adsorption followed by ATPE and ATPE followed by magnetic adsorption, for the separation of proteins mixture of lysozyme, bovine serum albumin, trypsin, cytochrome C and myloglobin were discussed and compared. The way of magnetic adsorption followed by ATPE was also applied to human serum separation. Copyright © 2011 Elsevier B.V. All rights reserved.

  13. Minicomputer system for radiochemical analysis by coincidence spectrometry

    International Nuclear Information System (INIS)

    Brauer, F.P.; Fager, J.E.

    1979-01-01

    Minicomputer-based coincidence analysis methods have been developed for use in performing radiochemical analysis by high-resolution x- and gamma-ray coincidence spectrometry. This paper describes the data-acquisition and analysis methods develolped for qualitative and quantitative analyses of coincidence spectrometric data. Data-acquisition capabilities include both direct multiparameter pulse-height analysis and buffered list-mode acquisition

  14. Instrumentation for chemical and radiochemical monitoring in nuclear power plants

    International Nuclear Information System (INIS)

    Nordmann, F.; Ballard, G.

    2009-01-01

    This article details the instrumentation implemented in French nuclear power plants for the monitoring of chemical and radiochemical effluents with the aim of limiting their environmental impact. It describes the controls performed with chemical automata for the search for drifts, anomalies or pollution in a given circuit. The operation principles of the different types of chemical automata are explained as well as the manual controls performed on samples manually collected. Content: 1 - general considerations; 2 - objectives of the chemical monitoring: usefulness of continuous monitoring with automata, transmission to control rooms and related actions, redundancy of automata; 3 - instrumentation and explanations for the main circuits: principle of chemical automata monitoring, instrumentation of the main primary circuit, instrumentation of the main secondary circuit, instrumentation of the tertiary circuit, preparation of water makeup (demineralized water), other loops, instrumentation for effluents and environment monitoring, measurement principles of chemical automata, control and maintenance of chemical automata; 4 - manual controls after sampling; 5 - radiochemical monitoring: automatized radiochemical measurements, manual radiochemical measurements; 6 - conclusion

  15. Surfactant protein A (SP-A) inhibits agglomeration and macrophage uptake of toxic amine modified nanoparticles.

    Science.gov (United States)

    McKenzie, Zofi; Kendall, Michaela; Mackay, Rose-Marie; Whitwell, Harry; Elgy, Christine; Ding, Ping; Mahajan, Sumeet; Morgan, Cliff; Griffiths, Mark; Clark, Howard; Madsen, Jens

    2015-01-01

    The lung provides the main route for nanomaterial exposure. Surfactant protein A (SP-A) is an important respiratory innate immune molecule with the ability to bind or opsonise pathogens to enhance phagocytic removal from the airways. We hypothesised that SP-A, like surfactant protein D, may interact with inhaled nanoparticulates, and that this interaction will be affected by nanoparticle (NP) surface characteristics. In this study, we characterise the interaction of SP-A with unmodified (U-PS) and amine-modified (A-PS) polystyrene particles of varying size and zeta potential using dynamic light scatter analysis. SP-A associated with both 100 nm U-PS and A-PS in a calcium-independent manner. SP-A induced significant calcium-dependent agglomeration of 100 nm U-PS NPs but resulted in calcium-independent inhibition of A-PS self agglomeration. SP-A enhanced uptake of 100 nm U-PS into macrophage-like RAW264.7 cells in a dose-dependent manner but in contrast inhibited A-PS uptake. Reduced association of A-PS particles in RAW264.7 cells following pre-incubation of SP-A was also observed with coherent anti-Stokes Raman spectroscopy. Consistent with these findings, alveolar macrophages (AMs) from SP-A(-/-) mice were more efficient at uptake of 100 nm A-PS compared with wild type C57Bl/6 macrophages. No difference in uptake was observed with 500 nm U-PS or A-PS particles. Pre-incubation with SP-A resulted in a significant decrease in uptake of 100 nm A-PS in macrophages isolated from both groups of mice. In contrast, increased uptake by AMs of U-PS was observed after pre-incubation with SP-A. Thus we have demonstrated that SP-A promotes uptake of non-toxic U-PS particles but inhibits the clearance of potentially toxic A-PS particles by blocking uptake into macrophages.

  16. Analysis of potential protein-modifying variants in 9000 endometriosis patients and 150000 controls of European ancestry

    DEFF Research Database (Denmark)

    Sapkota, Yadav; Vivo, Immaculata De; Steinthorsdottir, Valgerdur

    2017-01-01

    -modifying variants in endometriosis using exome-array genotyping in 7164 cases and 21005 controls, and a replication set of 1840 cases and 129016 controls of European ancestry. Results in the discovery sample identified significant evidence for association with coding variants in single-variant (rs1801232-CUBN...... sufficient power, our results did not identify any protein-modifying variants (MAF > 0.01) with moderate or large effect sizes in endometriosis, although these variants may exist in non-European populations or in high-risk families. The results suggest continued discovery efforts should focus on genotyping...

  17. TaBoo SeArch Algorithm with a Modified Inverse Histogram for Reproducing Biologically Relevant Rare Events of Proteins.

    Science.gov (United States)

    Harada, Ryuhei; Takano, Yu; Shigeta, Yasuteru

    2016-05-10

    The TaBoo SeArch (TBSA) algorithm [ Harada et al. J. Comput. Chem. 2015 , 36 , 763 - 772 and Harada et al. Chem. Phys. Lett. 2015 , 630 , 68 - 75 ] was recently proposed as an enhanced conformational sampling method for reproducing biologically relevant rare events of a given protein. In TBSA, an inverse histogram of the original distribution, mapped onto a set of reaction coordinates, is constructed from trajectories obtained by multiple short-time molecular dynamics (MD) simulations. Rarely occurring states of a given protein are statistically selected as new initial states based on the inverse histogram, and resampling is performed by restarting the MD simulations from the new initial states to promote the conformational transition. In this process, the definition of the inverse histogram, which characterizes the rarely occurring states, is crucial for the efficiency of TBSA. In this study, we propose a simple modification of the inverse histogram to further accelerate the convergence of TBSA. As demonstrations of the modified TBSA, we applied it to (a) hydrogen bonding rearrangements of Met-enkephalin, (b) large-amplitude domain motions of Glutamine-Binding Protein, and (c) folding processes of the B domain of Staphylococcus aureus Protein A. All demonstrations numerically proved that the modified TBSA reproduced these biologically relevant rare events with nanosecond-order simulation times, although a set of microsecond-order, canonical MD simulations failed to reproduce the rare events, indicating the high efficiency of the modified TBSA.

  18. Modifying a standard method allows simultaneous extraction of RNA and protein, enabling detection of enzymes in the rat retina with low expressions and protein levels.

    Science.gov (United States)

    Agardh, Elisabet; Gustavsson, Carin; Hagert, Per; Nilsson, Marie; Agardh, Carl-David

    2006-02-01

    The aim of the study was to evaluate messenger RNA and protein expression in limited amounts of tissue with low protein content. The Chomczynski method was used for simultaneous extraction of RNA, and protein was modified in the protein isolation step. Template mass and cycling time for the complementary DNA synthesis step of real-time reverse transcription-polymerase chain reaction (RT-PCR) for analysis of catalase, copper/zinc superoxide dismutase, manganese superoxide dismutase, the catalytic subunit of glutamylcysteine ligase, glutathione peroxidase 1, and the endogenous control cyclophilin B (CypB) were optimized before PCR. Polymerase chain reaction accuracy and efficacy were demonstrated by calculating the regression (R2) values of the separate amplification curves. Appropriate antibodies, blocking buffers, and running conditions were established for Western blot, and protein detection and multiplex assays with CypB were performed for each target. During the extraction procedure, the protein phase was dissolved in a modified washing buffer containing 0.1% sodium dodecyl sulfate, followed by ultrafiltration. Enzyme expression on real-time RT-PCR was accomplished with high reliability and reproducibility (R2, 0.990-0.999), and all enzymes except for glutathione peroxidase 1 were detectable in individual retinas on Western blot. Western blot multiplexing with CypB was possible for all targets. In conclusion, connecting gene expression directly to protein levels in the individual rat retina was possible by simultaneous extraction of RNA and protein. Real-time RT-PCR and Western blot allowed accurate detection of retinal protein expressions and levels.

  19. Present status and perspective of radiochemical analysis of radionuclides in Nordic countries

    DEFF Research Database (Denmark)

    Hou, Xiaolin; Olsson, Mattias; Togneri, Laura

    2016-01-01

    Radiochemical analysis plays a critical role in the determination of pure beta and alpha emitting radionuclides for environmental monitoring, radioecology, decommissioning, nuclear forensics and geological dating. A remarkable development on radiochemical analysis has been achieved in the past...... of radionuclides, especially in Nordic countries; some requirements from nuclear industries and research organizations, as well as perspectives on the development of radiochemical analysis are discussed....

  20. Radiochemical guidelines and process specifications for reactor shutdown: the EDF strategy

    International Nuclear Information System (INIS)

    Mole, D.; Wintergerst, M.; Meylogan, Th.; Rocher, A.; Sagot, M.J.; Bonelli, V.; Bonnefon, J.; Dupont, B.

    2012-09-01

    Changes to French nuclear regulations made in June 2006 [1.] have made it necessary for EDF to modify its ruling principles. These modifications required the restructuring of radiochemical guidelines to better reflect their impact on nuclear safety, the environment and radioprotection. In accordance with these aims, a new authoritative document has been produced. This ruling document identifies all parameters with a potential impact on nuclear safety, radiological releases to the environment and personnel dose rates. These diagnostic and control parameters have been identified for a reactor in production and for a reactor during shutdown. For parameters related to a reactor in production, some indicators are used to evaluate impacts on availability, radioprotection and the environment during shutdown and on outage and to anticipate mitigation ways. On the other side, several parameters related to the stages of shutdown were also directly evaluated in order to minimize the impacts. This paper describes the EDF methodology used to establish operational documents: radiochemical guidelines and process specifications, and includes the following: - description of monitored parameters and their associated areas of risk; - justification of target values, frequencies of inspection and the required actions for the monitored parameters. The sizing methodology is based on theoretical studies and on EDF operational experience analysis. By implementing in the operational and technical specifications requirements linked to nuclear safety, radioprotection and environment respect, EDF will benefit from an improved compromise between these areas as well as an increased focus. (authors)

  1. The mechanism of lauric acid-modified protein nanocapsules escape from intercellular trafficking vesicles and its implication for drug delivery.

    Science.gov (United States)

    Jiang, Lijuan; Liang, Xin; Liu, Gan; Zhou, Yun; Ye, Xinyu; Chen, Xiuli; Miao, Qianwei; Gao, Li; Zhang, Xudong; Mei, Lin

    2018-11-01

    Protein nanocapsules have exhibited promising potential applications in the field of protein drug delivery. A major issue with various promising nano-sized biotherapeutics including protein nanocapsules is that owing to their particle size they are subject to cellular uptake via endocytosis, and become entrapped and then degraded within endolysosomes, which can significantly impair their therapeutic efficacy. In addition, many nano-sized biotherapeutics could be also sequestered by autophagosomes and degraded through the autolysosomal pathway. Thus, a limiting step in achieving an effective protein therapy is to facilitate the endosomal escape and auto-lysosomal escape to ensure cytosolic delivery of the protein drugs. Here, we prepared a protein nanocapsule based on BSA (nBSA) and the BSA nanocapsules modified with a bilayer of lauric acid (LA-nBSA) to investigate the escape effects from the endosome and autophagosome. The size distribution of nBSA and LA-nBSA analyzed using DLS presents a uniform diameter centered at 10 nm and 16 nm. The data also showed that FITC-labeled nBSA and LA-nBSA were taken up by the cells mainly through Arf-6-dependent endocytosis and Rab34-mediated macropinocytosis. In addition, LA-nBSA could efficiently escape from endosomal before the degradation in endo-lysosomes. Autophagy could also sequester the LA-nBSA through p62 autophagosome vesicles. These two types of nanocapsules underwent different intracellular destinies and lauric acid (LA) coating played a vital role in intracellular particle retention. In conclusion, the protein nanocapsules modified with LA could enhance the protein nanocapsules escape from intercellular trafficking vesicles, and protect the protein from degradation by the lysosomes.

  2. Protein-Modified-Paramagnetic-Particles as a Tool for Detection of Silver(I) Ions

    Science.gov (United States)

    Kizek, R.; Krizkova, S.; Adam, V.; Huska, D.; Hubalek, J.; Trnkova, L.

    2009-04-01

    incorporated mixed with carbon paste were stable. Even after 14 days we did not determine change in the peak height higher than 5 %. Further linkage of MT with polyclonal chicken antibodies incorporated in carbon paste electrode was determined by SWV. Two signals were observed in voltammograms, cysMT corresponding to -SH moieties of MT and Wa corresponding to tryptophan residues of chicken antibodies. We optimized time of interaction (300 s) and concentration of MT (125 µg/ml) to suggest silver(I) ions biosensor. Biosensor (MT-antibody-modified CPE) prepared under the optimized conditions was utilized for silver(I) ions detection. The detection limit (3 S/N) for silver(I) ions were estimated as 100 nM. The proposed biosensor was tested by detection of silver(I) ions spiked in various water samples (from very pure distilled water to rainwater). Recoveries varied from 74 to 104 %. MT, low molecular mass proteins rich cysteine, play important role in the processes of heavy metals ions metabolism. Due to their unique physico-chemical properties they are able to bind heavy metals with high affinity. We used this feature to suggest simple biosensor based on immobilization of MT on the surface of carbon paste electrode via chicken antibodies against MT. The suggested biosensor was further successfully employed to detect silver(I) ions. The main advantage of the biosensor is that it can be easily miniaturized, whereas carbon nanostructures with immobilized MT should be used as working electrodes. Acknowledgements Financial support from INCHEMBIOL MSMT 0021622412 and GA CR 526/07/0674 is highly acknowledged.

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

    Directory of Open Access Journals (Sweden)

    Akinobu Kajikawa

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

  4. Toxicity assessment of modified Cry1Ac1 proteins and genetically ...

    African Journals Online (AJOL)

    Owner

    2015-06-10

    Jun 10, 2015 ... Key words: Modified Cry1Ac1, food safety assessment, toxicity, insect- resistant rice Agb0101. INTRODUCTION. Genetically modified (GM) crops are becoming an increasingly important feature of the agricultural land- scapes. In 2013, approximately 175 million hectares of. GM crops were planted by 18 ...

  5. Protein adsorption resistance of PVP-modified polyurethane film prepared by surface-initiated atom transfer radical polymerization

    Energy Technology Data Exchange (ETDEWEB)

    Yuan, Huihui; Qian, Bin; Zhang, Wei [Shanghai Key Laboratory of Functional Materials Chemistry and Research Center of Analysis and Test, East China University of Science and Technology, Shanghai 200237 (China); Lan, Minbo, E-mail: minbolan@ecust.edu.cn [Shanghai Key Laboratory of Functional Materials Chemistry and Research Center of Analysis and Test, East China University of Science and Technology, Shanghai 200237 (China); State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237 (China)

    2016-02-15

    Highlights: • Antifouling PVP brushes were successfully grafted on PU films by SI-ATRP. • The effect of polymerization time on surface property and topography was studied. • Hydrophilicity and protein fouling resistance of PVP–PU films were greatly promoted. • Competitive adsorption of three proteins on PVP–PU films was evaluated. - Abstract: An anti-fouling surface of polyurethane (PU) film grafted with Poly(N-vinylpyrrolidone) (PVP) was prepared through surface-initiated atom transfer radical polymerization (SI-ATRP). And the polymerization time was investigated to obtain PU films with PVP brushes of different lengths. The surface properties and protein adsorption of modified PU films were evaluated. The results showed that the hydrophilicity of PU–PVP films were improved with the increase of polymerization time, which was not positive correlation with the surface roughness due to the brush structure. Additionally, the protein resistance performance was promoted when prolonging the polymerization time. The best antifouling PU–PVP (6.0 h) film reduced the adsoption level of bovine serum albumin (BSA), lysozyme (LYS), and brovin serum fibrinogen (BFG) by 93.4%, 68.3%, 85.6%, respectively, compared to the unmodified PU film. The competitive adsorption of three proteins indicated that LYS preferentially adsorbed on the modified PU film, while BFG had the lowest adsorption selectivity. And the amount of BFG on PU–PVP (6.0 h) film reduced greatly to 0.08 μg/cm{sup 2}, which was almost one-tenth of its adsorption from the single-protein system. Presented results suggested that both hydrophilicity and surface roughness might be the important factors in all cases of protein adsorption, and the competitive or selective adsorption might be related to the size of the proteins, especially on the non-charged films.

  6. Protein adsorption resistance of PVP-modified polyurethane film prepared by surface-initiated atom transfer radical polymerization

    International Nuclear Information System (INIS)

    Yuan, Huihui; Qian, Bin; Zhang, Wei; Lan, Minbo

    2016-01-01

    Highlights: • Antifouling PVP brushes were successfully grafted on PU films by SI-ATRP. • The effect of polymerization time on surface property and topography was studied. • Hydrophilicity and protein fouling resistance of PVP–PU films were greatly promoted. • Competitive adsorption of three proteins on PVP–PU films was evaluated. - Abstract: An anti-fouling surface of polyurethane (PU) film grafted with Poly(N-vinylpyrrolidone) (PVP) was prepared through surface-initiated atom transfer radical polymerization (SI-ATRP). And the polymerization time was investigated to obtain PU films with PVP brushes of different lengths. The surface properties and protein adsorption of modified PU films were evaluated. The results showed that the hydrophilicity of PU–PVP films were improved with the increase of polymerization time, which was not positive correlation with the surface roughness due to the brush structure. Additionally, the protein resistance performance was promoted when prolonging the polymerization time. The best antifouling PU–PVP (6.0 h) film reduced the adsoption level of bovine serum albumin (BSA), lysozyme (LYS), and brovin serum fibrinogen (BFG) by 93.4%, 68.3%, 85.6%, respectively, compared to the unmodified PU film. The competitive adsorption of three proteins indicated that LYS preferentially adsorbed on the modified PU film, while BFG had the lowest adsorption selectivity. And the amount of BFG on PU–PVP (6.0 h) film reduced greatly to 0.08 μg/cm"2, which was almost one-tenth of its adsorption from the single-protein system. Presented results suggested that both hydrophilicity and surface roughness might be the important factors in all cases of protein adsorption, and the competitive or selective adsorption might be related to the size of the proteins, especially on the non-charged films.

  7. Colon luminal content and epithelial cell morphology are markedly modified in rats fed with a high-protein diet

    OpenAIRE

    Andriamihaja, Mireille; Davila-Gay, Anne-Marie; Eklou, Mamy; Petit, Nathalie; Delpal, Serge; Allek, Fadhila; Blais, Anne; Delteil, Corine; Tomé, Daniel; Blachier, Francois

    2010-01-01

    Andriamihaja M, Davila A, Eklou-Lawson M, Petit N, Delpal S, Allek F, Blais A, Delteil C, Tome D, Blachier F. Colon luminal content and epithelial cell morphology are markedly modified in rats fed with a high-protein diet. Am J Physiol Gastrointest Liver Physiol 299: G1030-G1037, 2010. First published August 5, 2010; doi: 10.1152/ajpgi.00149.2010.-Hyperproteic diets are used in human nutrition to obtain body weight reduction. Although increased protein ingestion results in an increased transf...

  8. The Role of Histone Protein Modifications and Mutations in Histone Modifiers in Pediatric B-Cell Progenitor Acute Lymphoblastic Leukemia

    Science.gov (United States)

    Janczar, Szymon; Janczar, Karolina; Pastorczak, Agata; Harb, Hani; Paige, Adam J. W.; Zalewska-Szewczyk, Beata; Danilewicz, Marian; Mlynarski, Wojciech

    2017-01-01

    While cancer has been long recognized as a disease of the genome, the importance of epigenetic mechanisms in neoplasia was acknowledged more recently. The most active epigenetic marks are DNA methylation and histone protein modifications and they are involved in basic biological phenomena in every cell. Their role in tumorigenesis is stressed by recent unbiased large-scale studies providing evidence that several epigenetic modifiers are recurrently mutated or frequently dysregulated in multiple cancers. The interest in epigenetic marks is especially due to the fact that they are potentially reversible and thus druggable. In B-cell progenitor acute lymphoblastic leukemia (BCP-ALL) there is a relative paucity of reports on the role of histone protein modifications (acetylation, methylation, phosphorylation) as compared to acute myeloid leukemia, T-cell ALL, or other hematologic cancers, and in this setting chromatin modifications are relatively less well studied and reviewed than DNA methylation. In this paper, we discuss the biomarker associations and evidence for a driver role of dysregulated global and loci-specific histone marks, as well as mutations in epigenetic modifiers in BCP-ALL. Examples of chromatin modifiers recurrently mutated/disrupted in BCP-ALL and associated with disease outcomes include MLL1, CREBBP, NSD2, and SETD2. Altered histone marks and histone modifiers and readers may play a particular role in disease chemoresistance and relapse. We also suggest that epigenetic regulation of B-cell differentiation may have parallel roles in leukemogenesis. PMID:28054944

  9. Ethylene glycol assisted preparation of Ti(4+)-modified polydopamine coated magnetic particles with rough surface for capture of phosphorylated proteins.

    Science.gov (United States)

    Ma, Xiangdong; Ding, Chun; Yao, Xin; Jia, Li

    2016-07-27

    The reversible protein phosphorylation is very important in regulating almost all aspects of cell life, while the enrichment of phosphorylated proteins still remains a technical challenge. In this work, polydopamine (PDA) modified magnetic particles with rough surface (rPDA@Fe3O4) were synthesized by introduction of ethylene glycol in aqueous solution. The PDA coating possessing a wealth of catechol hydroxyl groups could serve as an active medium to immobilize titanium ions through the metal-catechol chelation, which makes the fabrication of titanium ions modified rPDA@Fe3O4 particles (Ti(4+)-rPDA@Fe3O4) simple and very convenient. The spherical Ti(4+)-rPDA@Fe3O4 particles have a surface area of 37.7 m(2) g(-1) and superparamagnetism with a saturation magnetization value of 38.4 emu g(-1). The amount of Ti element in the particle was measured to be 3.93%. And the particles demonstrated good water dispersibility. The particles were used as adsorbents for capture of phosphorylated proteins and they demonstrated affinity and specificity for phosphorylated proteins due to the specific binding sites (Ti(4+)). Factors affecting the adsorption of phosphorylated proteins on Ti(4+)-rPDA@Fe3O4 particles were investigated. The adsorption capacity of Ti(4+)-rPDA@Fe3O4 particles for κ-casein was 1105.6 mg g(-1). Furthermore, the particles were successfully applied to isolate phosphorylated proteins in milk samples, which demonstrated that Ti(4+)-rPDA@Fe3O4 particles had potential application in selective separation of phosphorylated proteins. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. Major membrane surface proteins of Mycoplasma hyopneumoniae selectively modified by covalently bound lipid

    International Nuclear Information System (INIS)

    Wise, K.S.; Kim, M.F.

    1987-01-01

    Surface protein antigens of Mycoplasma hyopneumoniae were identified by direct antibody-surface binding or by radioimmunoprecipitation of surface 125 I-labeled proteins with a series of monoclonal antibodies (MAbs). Radioimmunoprecipitation of TX-114-phase proteins from cells labeled with [ 35 S] methionine, 14 C-amino acids, or [ 3 H] palmitic acid showed that proteins p65, p50, and p44 were abundant and (with one other hydrophobic protein, p60) were selectively labeled with lipid. Alkaline hydroxylamine treatment of labeled proteins indicated linkage of lipids by amide or stable O-linked ester bonds. Proteins p65, p50, and p44 were highly immunogenic in the natural host as measured by immunoblots of TX-114-phase proteins with antisera from swine inoculated with whole organisms. These proteins were antigenically and structurally unrelated, since hyperimmune mouse antibodies to individual gel-purified proteins were monospecific and gave distinct proteolytic epitope maps. Intraspecies size variants of one surface antigen of M. hyopneumoniae were revealed by a MAb to p70 (defined in strain J, ATCC 25934), which recognized a large p73 component on strain VPP11 (ATCC 25617). In addition, MAb to internal, aqueous-phase protein p82 of strain J failed to bind an analogous antigen in strain VPP11

  11. Major membrane surface proteins of Mycoplasma hyopneumoniae selectively modified by covalently bound lipid

    Energy Technology Data Exchange (ETDEWEB)

    Wise K.S.; Kim, M.F.

    1987-12-01

    Surface protein antigens of Mycoplasma hyopneumoniae were identified by direct antibody-surface binding or by radioimmunoprecipitation of surface /sup 125/I-labeled proteins with a series of monoclonal antibodies (MAbs). Radioimmunoprecipitation of TX-114-phase proteins from cells labeled with (/sup 35/S) methionine, /sup 14/C-amino acids, or (/sup 3/H) palmitic acid showed that proteins p65, p50, and p44 were abundant and (with one other hydrophobic protein, p60) were selectively labeled with lipid. Alkaline hydroxylamine treatment of labeled proteins indicated linkage of lipids by amide or stable O-linked ester bonds. Proteins p65, p50, and p44 were highly immunogenic in the natural host as measured by immunoblots of TX-114-phase proteins with antisera from swine inoculated with whole organisms. These proteins were antigenically and structurally unrelated, since hyperimmune mouse antibodies to individual gel-purified proteins were monospecific and gave distinct proteolytic epitope maps. Intraspecies size variants of one surface antigen of M. hyopneumoniae were revealed by a MAb to p70 (defined in strain J, ATCC 25934), which recognized a large p73 component on strain VPP11 (ATCC 25617). In addition, MAb to internal, aqueous-phase protein p82 of strain J failed to bind an analogous antigen in strain VPP11.

  12. Miniaturized chromatographic radiochemical procedure for 131I - MIBG

    International Nuclear Information System (INIS)

    Barboza, M.F. de; Pereira, N.S. de; Colturato, M.T.; Silva, C.P.G. da.

    1989-12-01

    Different solvents were used in paper chromatographic methods to obtain the best system in routine radiochemical control for 131 I-MIBG produced at IPEN-CNEN/SP. The dates were compared with those obtained with eletrophoresis method in buffer acetate, pH=4.5, 350V, during 40 minutes. The stability of the labeled compound store under 4 0 C was studied during 15 days. Miniaturized chromatographic procedures were established using Whatman 3MM (8x1cm) and n-butanol-:acetic acid: water (S:2:1) as a solvent. the Rf values were: 0.3 (I - ) and 1.0 (MIBG). The radiochemical purity was 99.3 and 99.2% (first day) obtained with eletrophoresis and miniaturized chromatographic procedures, respectively and, 84.7% after 15 days of its preparation. It is a rapid, practical and reproductive method. (author) [pt

  13. Thorium base fuels reprocessing at the L.P.R. (Radiochemical Processes Laboratory) experimental plant

    International Nuclear Information System (INIS)

    Almagro, J.C.; Dupetit, G.A.; Deandreis, R.A.

    1987-01-01

    The availability of the LPR (Radiochemical Processes Laboratory) plant offers the possibility to demonstrate and create the necessary technological basis for thorium fuels reprocessing. To this purpose, the solvents extraction technique is used, employing TBP (at 30%) as solvent. The process is named THOREX, a one-cycle acid, which permits an adequate separation of Th 232 and U 233 components and fission products. For thorium oxide elements dissolution, the 'chopp-leach' process (installed at LPR) is used, employing a NO 3 H 13N, 0.05M FH and 0.1M Al (NO 3 ) 3 , as solvent. To adapt the pilot plant to the flow-sheet requirements proposed, minor modifications must be carried out in the interconnection of the existing decanting mixers. The input of the plant has been calculated by Origin Code modified for irradiations in reactors of the HWR type. (Author)

  14. Rapid radiochemical separation of zirconium-95 and niobium-95

    International Nuclear Information System (INIS)

    Downey, D.M.; McLaughlin, C.L.

    1983-01-01

    A rapid method for the quantitative separation of 95 Zr and 95 Nb has been developed. The method is based on the ion flotation of cationic zirconium complex ions with sodium lauryl sulfate (NaLS) from niobium which is masked with hydrogen peroxide. The separation was applied to mixtures of 95 Zr and 95 Nb initially in oxalic acid solution and quantitative recoveries of the radiochemically pure radioisotopes were obtained. (orig.)

  15. Rapid radiochemical separation of zirconium-95 and niobium-95

    Energy Technology Data Exchange (ETDEWEB)

    Downey, D.M.; McLaughlin, C.L.

    1983-01-01

    A rapid method for the quantitative separation of /sup 95/Zr and /sup 95/Nb has been developed. The method is based on the ion flotation of cationic zirconium complex ions with sodium lauryl sulfate (NaLS) from niobium which is masked with hydrogen peroxide. The separation was applied to mixtures of /sup 95/Zr and /sup 95/Nb initially in oxalic acid solution and quantitative recoveries of the radiochemically pure radioisotopes were obtained.

  16. Radiochemical regularities of migration mobility of Chernobyl' discharge radionuclides

    International Nuclear Information System (INIS)

    Skorobogat'ko, E.P.; Rybalko, S.I.

    1992-01-01

    Data on the radionuclude (RN) migration in environment later the Chernobyl' accident are generalized. Introduction of fallout of the radioactive discharge into environment causes necessity to account and to study different factors of geochemical and physicochemical character determining further RN behaviour in the medium. For a well-founded forecast of the behaviour it is necessity to use a complex of radiochemical and physicochemical research, lying in the base of radiation monitoring of environment. 1 refs

  17. An overview of the status of radiochemical analysis in Hungary

    International Nuclear Information System (INIS)

    Solymosi, J.; Toth, G.

    1994-01-01

    This overview covers the following activities at radioanalytical laboratories in Hungary: tracer techniques and their applications; some important new results; radioimmunoassay; x-ray emission analysis and x-ray fluorescence analysis, Moessbauer-spectroscopy and their applications in various fields of science and technology; neutron activation analysis; radiochemical analysis for nuclear power plant applications activities in various laboratories; nuclear environmental analysis (radioanalytical methods for the investigation of contamination by nuclear facilities). (N.T.) 1 fig.; 7 tabs

  18. Electrochemistry as a basis for radiochemical generator systems

    International Nuclear Information System (INIS)

    Bentley, G.E.; Steinkruger, F.J.; Wanek, P.M.

    1984-01-01

    Ion exchange and solvent extraction techniques have been used extensively as the basis for radiochemical generators exploiting the differences in absorption behavior between the parent nuclide and its useful daughter nuclide. Many parent/daughter pairs of nuclides have sufficiently different polarographic half wave potentials so that their electrochemical behavior may be exploited for rapid separation of the daughter from the parent with minimal contamination of the product with the parent isotope

  19. Decontamination and decommission of a radiochemical laboratory building complex

    International Nuclear Information System (INIS)

    Zoubek, Norbert

    2008-01-01

    Full text: Handling of unsealed radioactive substances for research and development purposes in chemical or pharmaceutical industries or research centres as well as production of radioactive substances (e.g. for applications in nuclear medicine or industry) requires operation of special radiochemical laboratories. In general, operation of radiochemical laboratories is strongly regulated by the government and national authorities. The operator needs a permit related to radiological protection. In general, technical requirements for such facilities are very high. To ensure high safety standards with respect to the employees and the environment, several radiological protection measures have to be taken. These measures (for example special shielding or ventilation and waste water systems) depend on various factors, e.g. activity in use, kind of nuclides, chemical properties and volatility of substances. In order to close-down such radiochemical laboratories some radiological protection measures have to be maintained to ensure protection of both humans and the environment induced by possible residual contaminations within the facility including technical inventory. However, a later reuse of the facility as a non-radioactive facility requires removal of all radioactive contamination with respect to national regulation. Resulting radioactive wastes have to be disposed of under control of competent authorities. Based on the experience of a decontamination and decommission project for a former radiochemical laboratory complex, the main steps necessary to release such a facility are discussed. Analytical aspects of initial conditions, necessary organisational structures within the project, resources needed estimation and exploration of the radiological situation in the laboratory, elaboration of a measuring strategy and decontamination methods as well as different waste disposal routes in relation to different waste types are reported. (author)

  20. Present status of radiochemical double β decay study (238U)

    International Nuclear Information System (INIS)

    Chevallier, A.; Chevallier, J.; Escoubes, B.; Schulz, N.; Sens, J.C.; Madic, C.; Maillard, C.

    1989-01-01

    The reasons for which the 238 U is a suitable candidate for the β β decay processes are explained. The strategy adopted for the radiochemical separation of the 234 U is given. A chemical system based on extraction chromatography is applied. The Pu IV breakthrough curves obtained at 40C during 238 Pu/ 238 U separation cycles are presented. A short description of the chromatographic facility is given. The solution adopted for the low background α spectrometer is explained

  1. Radiochemical separation and their application to neutron activation analysis technique

    International Nuclear Information System (INIS)

    Turel, Z.R.

    2013-01-01

    The present paper discusses the development of some new, rapid and selective method for the radiochemical separation and estimation of elements such as, Co(II) 2-3 , Ir(III) 4 , Au(III) 5 , Pt(IV), Pd(II), Os(IV) 6 , Cu(II), Ag(I), Mo(VI), Ni(II), Zn(II), Cd(II), Hg(II), Cs(I), Sb(III), La(III), Sc(III) etc. using various reagents. Various parameters such as pH, time of equilibrium, effect of anions and cations, effect of reagent etc. has been determined employing tracers of the elements under consideration and will be discussed. The method is made highly selective by the use of appropriate masking agent. The stoichiometry of metal reagent is determined by the substoichiometric method. Some examples of multielemental radiochemical separation methods thus developed which have been applied in determining the elements by radiochemical thermal neutron activation analysis will be presented and discussed. The implications of the results on the reference system will also be accounted. Statistical evaluation with reference to accuracy, precision and sensitivity will also be presented

  2. A modified strategy for sequence specific assignment of protein NMR spectra based on amino acid type selective experiments

    International Nuclear Information System (INIS)

    Schubert, Mario; Labudde, Dirk; Leitner, Dietmar; Oschkinat, Hartmut; Schmieder, Peter

    2005-01-01

    The determination of the three-dimensional structure of a protein or the study of protein-ligand interactions requires the assignment of all relevant nuclei as an initial step. This is nowadays almost exclusively performed using triple-resonance experiments. The conventional strategy utilizes one or more pairs of three dimensional spectra to obtain redundant information and thus reliable assignments. Here, a modified strategy for obtaining sequence specific assignments based on two dimensional amino acid type selective triple-resonance experiments is proposed. These experiments can be recorded with good resolution in a relatively short time. They provide very specific and redundant information, in particular on sequential connectivities, that drastically increases the ease and reliability of the assignment procedure, done either manually or in an automated fashion. The new strategy is demonstrated with the protein domain PB1 from yeast CDC24p

  3. Physicochemical and Antioxidant Properties of Buckwheat Protein Isolates with Different Polyphenolic Content Modified by Limited Hydrolysis with Trypsin

    Directory of Open Access Journals (Sweden)

    Xiao-Yan Wang

    2012-01-01

    Full Text Available Effects of limited hydrolysis with trypsin on the physicochemical and antioxidant properties of buckwheat protein isolates (BPIs obtained with untreated and 2-propanol-extracted meal have been investigated and compared. The dephenolization treatment significantly improved the hydrolysis of BPI, which resulted in the gradual decrease in total and protein-bound polyphenolic content, but an increase in the free polyphenolic content. The hydrolysis of globulins was much easier than that of the albumins. The removal of polyphenols improved the hydrolysis of the albumin fraction. The modified BPIs with high polyphenolic content exhibited much higher DPPH radical scavenging activity and reducing power, but poorer ferrous ion chelating ability than those with low polyphenolic content. These results suggest that the limited hydrolysis is suitable for modification of the properties of buckwheat proteins.

  4. Pepsin immobilized in dextran-modified fused-silica capillaries for on-line protein digestion and peptide mapping.

    Science.gov (United States)

    Stigter, E C A; de Jong, G J; van Bennekom, W P

    2008-07-07

    On-line digestion of proteins under acidic conditions was studied using micro-reactors consisting of dextran-modified fused-silica capillaries with covalently immobilized pepsin. The proteins used in this study differed in molecular weight, isoelectric point and sample composition. The injected protein samples were completely digested in 3 min and the digest was analyzed with micro-high performance liquid chromatography (HPLC) and tandem mass spectrometry (MS/MS). The different proteins present in the samples could be identified with a Mascot database search on the basis of auto-MS/MS data. It proved also to be possible to digest and analyze protein mixtures with a sequence coverage of 55% and 97% for the haemoglobin beta- and alpha-chain, respectively, and 35-55% for the various casein variants. Protease auto-digestion, sample carry-over and loss of signal due to adsorption of the injected proteins were not observed. The backpressure of the reactor is low which makes coupling to systems such as Surface Plasmon Resonance biosensors, which do not tolerate too high pressure, possible. The reactor was stable for at least 40 days when used continuously.

  5. Pepsin immobilized in dextran-modified fused-silica capillaries for on-line protein digestion and peptide mapping

    Energy Technology Data Exchange (ETDEWEB)

    Stigter, E.C.A. [Division of Biomedical Analysis, Department of Pharmaceutical Sciences, Faculty of Science, Utrecht University, Sorbonnelaan 16, 3584 CA Utrecht (Netherlands)], E-mail: e.c.a.stigter@uu.nl; Jong, G.J. de; Bennekom, W.P. van [Division of Biomedical Analysis, Department of Pharmaceutical Sciences, Faculty of Science, Utrecht University, Sorbonnelaan 16, 3584 CA Utrecht (Netherlands)

    2008-07-07

    On-line digestion of proteins under acidic conditions was studied using micro-reactors consisting of dextran-modified fused-silica capillaries with covalently immobilized pepsin. The proteins used in this study differed in molecular weight, isoelectric point and sample composition. The injected protein samples were completely digested in 3 min and the digest was analyzed with micro-high performance liquid chromatography (HPLC) and tandem mass spectrometry (MS/MS). The different proteins present in the samples could be identified with a Mascot database search on the basis of auto-MS/MS data. It proved also to be possible to digest and analyze protein mixtures with a sequence coverage of 55% and 97% for the haemoglobin {beta}- and {alpha}-chain, respectively, and 35-55% for the various casein variants. Protease auto-digestion, sample carry-over and loss of signal due to adsorption of the injected proteins were not observed. The backpressure of the reactor is low which makes coupling to systems such as Surface Plasmon Resonance biosensors, which do not tolerate too high pressure, possible. The reactor was stable for at least 40 days when used continuously.

  6. Protein resistance of surfaces modified with oligo(ethylene glycol) aryl diazonium derivatives.

    Science.gov (United States)

    Fairman, Callie; Ginges, Joshua Z; Lowe, Stuart B; Gooding, J Justin

    2013-07-22

    Anti-fouling surfaces are of great importance for reducing background interference in biosensor signals. Oligo(ethylene glycol) (OEG) moieties are commonly used to confer protein resistance on gold, silicon and carbon surfaces. Herein, we report the modification of surfaces using electrochemical deposition of OEG aryl diazonium salts. Using electrochemical and contact angle measurements, the ligand packing density is found to be loose, which supports the findings of the fluorescent protein labelling that aryl diazonium OEGs confer resistance to nonspecific adsorption of proteins albeit lower than alkane thiol-terminated OEGs. In addition to protein resistance, aryl diazonium attachment chemistry results in stable modification. In common with OEG species on gold electrodes, OEGs with distal hydroxyl moieties do confer superior protein resistance to those with a distal methoxy group. This is especially the case for longer derivatives where superior coiling of the OEG chains is possible. Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  7. Receptor Activity-modifying Protein-directed G Protein Signaling Specificity for the Calcitonin Gene-related Peptide Family of Receptors.

    Science.gov (United States)

    Weston, Cathryn; Winfield, Ian; Harris, Matthew; Hodgson, Rose; Shah, Archna; Dowell, Simon J; Mobarec, Juan Carlos; Woodlock, David A; Reynolds, Christopher A; Poyner, David R; Watkins, Harriet A; Ladds, Graham

    2016-10-14

    The calcitonin gene-related peptide (CGRP) family of G protein-coupled receptors (GPCRs) is formed through the association of the calcitonin receptor-like receptor (CLR) and one of three receptor activity-modifying proteins (RAMPs). Binding of one of the three peptide ligands, CGRP, adrenomedullin (AM), and intermedin/adrenomedullin 2 (AM2), is well known to result in a Gα s -mediated increase in cAMP. Here we used modified yeast strains that couple receptor activation to cell growth, via chimeric yeast/Gα subunits, and HEK-293 cells to characterize the effect of different RAMP and ligand combinations on this pathway. We not only demonstrate functional couplings to both Gα s and Gα q but also identify a Gα i component to CLR signaling in both yeast and HEK-293 cells, which is absent in HEK-293S cells. We show that the CGRP family of receptors displays both ligand- and RAMP-dependent signaling bias among the Gα s , Gα i , and Gα q/11 pathways. The results are discussed in the context of RAMP interactions probed through molecular modeling and molecular dynamics simulations of the RAMP-GPCR-G protein complexes. This study further highlights the importance of RAMPs to CLR pharmacology and to bias in general, as well as identifying the importance of choosing an appropriate model system for the study of GPCR pharmacology. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  8. Expression of PAT and NPT II proteins during the developmental stages of a genetically modified pepper developed in Korea.

    Science.gov (United States)

    Kim, Hyo Jin; Lee, Si Myung; Kim, Jae Kwang; Ryu, Tae Hun; Suh, Seok Cheol; Cho, Hyun Suk

    2010-10-27

    Estimation of the protein levels introduced in a biotechnology-derived product is conducted as part of an overall safety assessment. An enzyme-linked immunosorbent assay (ELISA) was used to analyze phosphinothricin acetyltransferase (PAT) and neomycin phosphotransferase II (NPT II) protein expression in a genetically modified (GM) pepper plant developed in Korea. PAT and NPT II expression levels, based on both dry weight and fresh weight, were variable among different plant generations and plant sections from isolated genetically modified organism (GMO) fields at four developmental stages. PAT expression was highest in leaves at anthesis (11.44 μg/gdw and 2.17 μg/gfw) and lowest in roots (0.12 μg/gdw and 0.01 μg/gfw). NPT II expression was also highest in leaves at anthesis (17.31 μg/gdw and 3.41 μg/gfw) and lowest in red pepper (0.65 μg/gdw and 0.12 μg/gfw). In pollen, PAT expression was 0.59-0.62 μg/gdw, while NPT II was not detected. Both PAT and NPT II showed a general pattern of decreased expression with progression of the growing season. As expected, PAT and NPT II protein expression was not detectable in control pepper plants.

  9. Biochemistry of plant class IV chitinases and fungal chitinase-modifying proteins

    Science.gov (United States)

    Plant class IV chitinases have 2 domains, a small (3 kDa) amino-terminal domain with homology to carbohydrate binding peptides, and a larger (25 kDa) catalytic domain. The biological function of these chitinases is not known. But it is known that some pathogenic fungi secrete chitinase modifying pro...

  10. Overexpression of the transcription factor Yap1 modifies intracellular redox conditions and enhances recombinant protein secretion

    Directory of Open Access Journals (Sweden)

    Marizela Delic

    2014-10-01

    Full Text Available Oxidative folding of secretory proteins in the endoplasmic reticulum (ER is a redox active process, which also impacts the redox conditions in the cytosol. As the transcription factor Yap1 is involved in the transcriptional response to oxidative stress, we investigate its role upon the production of secretory proteins, using the yeast Pichia pastoris as model, and report a novel important role of Yap1 during oxidative protein folding. Yap1 is needed for the detoxification of reactive oxygen species (ROS caused by increased oxidative protein folding. Constitutive co-overexpression of PpYAP1 leads to increased levels of secreted recombinant protein, while a lowered Yap1 function leads to accumulation of ROS and strong flocculation. Transcriptional analysis revealed that more than 150 genes were affected by overexpression of YAP1, in particular genes coding for antioxidant enzymes or involved in oxidation-reduction processes. By monitoring intracellular redox conditions within the cytosol and the ER using redox-sensitive roGFP1 variants, we could show that overexpression of YAP1 restores cellular redox conditions of protein-secreting P. pastoris by reoxidizing the cytosolic redox state to the levels of the wild type. These alterations are also reflected by increased levels of oxidized intracellular glutathione (GSSG in the YAP1 co-overexpressing strain. Taken together, these data indicate a strong impact of intracellular redox balance on the secretion of (recombinant proteins without affecting protein folding per se. Re-establishing suitable redox conditions by tuning the antioxidant capacity of the cell reduces metabolic load and cell stress caused by high oxidative protein folding load, thereby increasing the secretion capacity.

  11. IN-MACA-MCC: Integrated Multiple Attractor Cellular Automata with Modified Clonal Classifier for Human Protein Coding and Promoter Prediction

    Directory of Open Access Journals (Sweden)

    Kiran Sree Pokkuluri

    2014-01-01

    Full Text Available Protein coding and promoter region predictions are very important challenges of bioinformatics (Attwood and Teresa, 2000. The identification of these regions plays a crucial role in understanding the genes. Many novel computational and mathematical methods are introduced as well as existing methods that are getting refined for predicting both of the regions separately; still there is a scope for improvement. We propose a classifier that is built with MACA (multiple attractor cellular automata and MCC (modified clonal classifier to predict both regions with a single classifier. The proposed classifier is trained and tested with Fickett and Tung (1992 datasets for protein coding region prediction for DNA sequences of lengths 54, 108, and 162. This classifier is trained and tested with MMCRI datasets for protein coding region prediction for DNA sequences of lengths 252 and 354. The proposed classifier is trained and tested with promoter sequences from DBTSS (Yamashita et al., 2006 dataset and nonpromoters from EID (Saxonov et al., 2000 and UTRdb (Pesole et al., 2002 datasets. The proposed model can predict both regions with an average accuracy of 90.5% for promoter and 89.6% for protein coding region predictions. The specificity and sensitivity values of promoter and protein coding region predictions are 0.89 and 0.92, respectively.

  12. Manganese modified CdTe/CdS quantum dots as an immunoassay biosensor for the detection of Golgi protein-73.

    Science.gov (United States)

    Liu, Wei; Zhang, Aixia; Xu, Guanhong; Wei, Fangdi; Yang, Jing; Hu, Qin

    2016-01-05

    In this paper, a new fluorescence bioassay for Golgi protein-73 (GP73), a promising marker for monitoring liver tumor, was developed by using anti-GP73 antibody (GP73 Ab) capped quantum dots (QDs) coupled with protein A/G agarose beads in an attempt to improve the analysis time, cost and operation. First, carboxylic-functionalized Mn modified CdTe/CdS QDs were synthesized and covalently conjugated with GP73 Ab, then protein A/G agarose beads were specifically combined with the QDs-conjugated Ab to form the QDs-Ab-beads conjugate, which could capture and separate GP73 from the sample through simple centrifugation. It was found that the fluorescence intensity of the above QDs-Ab-beads biosensor could be specifically quenched by GP73 added. A simple, rapid and specific quantitative method for GP73 protein was proposed using the as-prepared QDs-Ab-beads as a biosensor. Under the optimized conditions, the calibration curve of the proposed assay showed good linearity with a correlation coefficient of 0.9935 in the concentration range of 20-150 ng/mL of GP73 protein. The limit of detection (defined as 3σ/K) was 10 ng/mL. The method built exhibited a great potential in the clinic test of GP73. Copyright © 2015 Elsevier B.V. All rights reserved.

  13. Characterization of Silk Fibroin Modified Surface: A Proteomic View of Cellular Response Proteins Induced by Biomaterials

    Directory of Open Access Journals (Sweden)

    Ming-Hui Yang

    2014-01-01

    Full Text Available The purpose of this study was to develop the pathway of silk fibroin (SF biopolymer surface induced cell membrane protein activation. Fibroblasts were used as an experimental model to evaluate the responses of cellular proteins induced by biopolymer material using a mass spectrometry-based profiling system. The surface was covered by multiwalled carbon nanotubes (CNTs and SF to increase the surface area, enhance the adhesion of biopolymer, and promote the rate of cell proliferation. The amount of adhered fibroblasts on CNTs/SF electrodes of quartz crystal microbalance (QCM greatly exceeded those on other surfaces. Moreover, analyzing differential protein expressions of adhered fibroblasts on the biopolymer surface by proteomic approaches indicated that CD44 may be a key protein. Through this study, utilization of mass spectrometry-based proteomics in evaluation of cell adhesion on biopolymer was proposed.

  14. Competitive Adsorption of Plasma Proteins on Polysaccharide-Modified Silicon Surfaces

    National Research Council Canada - National Science Library

    Ombelli, Michela; Costello, Lauren B; Meng, Qing C; Composto, Russell J; Eckmann, David M

    2005-01-01

    .... Competitive protein adsorption plays a key role in the hemocompatibility of the surface. The synthesis of nonfouling surfaces is therefore one of the major prerequisites for devices for biomedical applications...

  15. Analyzing modifiers of protein aggregation in C. elegans by native agarose gel electrophoresis

    NARCIS (Netherlands)

    Holmberg, Mats; Nollen, Ellen A A; Hatters, Danny M.; Hannan, Anthony J.

    2013-01-01

    The accumulation of specific aggregation-prone proteins during aging is thought to be involved in several diseases, most notably Alzheimer's and Parkinson's disease as well as polyglutamine expansion disorders such as Huntington's disease. Caenorhabditis elegans disease models with transgenic

  16. Modifier Genes for Mouse Phosphatidylinositol Transfer Protein alpha (vibrator) That Bypass Juvenile Lethality

    NARCIS (Netherlands)

    Concepcion, Dorothy; Johannes, Frank; Lo, Yuan Hung; Yao, Jay; Fong, Jerry; Hamilton, Bruce A.

    Phosphatidylinositol transfer proteins (PITPs) mediate lipid signaling and membrane trafficking in eukaryotic cells. Loss-of-function mutations of the gene encoding PITP alpha in mice result in a range of dosage-sensitive phenotypes, including neurological dysfunction, neurodegeneration, and

  17. Surface and protein analyses of normal human cell attachment on PIII-modified chitosan membranes

    International Nuclear Information System (INIS)

    Saranwong, N.; Inthanon, K.; Wongkham, W.; Wanichapichart, P.; Suwannakachorn, D.; Yu, L.D.

    2012-01-01

    Surface of chitosan membrane was modified with argon (Ar) and nitrogen (N) plasma immersion ion implantation (PIII) for human skin fibroblasts F1544 cell attachment. The modified surfaces were characterized by Fourier transform infrared spectroscopy (FTIR) and atomic force microscopy (AFM). Cell attachment patterns were evaluated by scanning electron microscopy (SEM). The enzyme-linked immunosorbent assay (ELISA) was used to quantify levels of focal adhesion kinase (FAK). The results showed that Ar PIII had an enhancement effect on the cell attachment while N-PIII had an inhibition effect. Filopodial analysis revealed more microfilament cytoplasmic spreading on the edge of cells attached on the Ar-treated membranes than N-treated membranes. Higher level FAK was found in Ar-treated membranes than that in N-treated membranes.

  18. Evolution and applications of radiochemical procedures. From Marie Curie to Darleane Hoffman

    International Nuclear Information System (INIS)

    Contis, E. T.; Rengan, K.

    1996-01-01

    Marie Curie carried out the first radiochemical separations which eventually lead to the discovery of polonium and radium, two new elements. Nearly a century later Darleane Hoffman and her collaborators are devising new radiochemical separation procedures for studying the chemical properties of newly discovered transactinide elements. Safety requirements as well as changes necessitated by fast decaying radionuclides have transformed the nature of radiochemical separations. Further, applications in a wide variety of areas such as analysis of trace elements in food to radioimmunoassay have broadened the use of radiochemical separations. Examples of some early, historically important, radiochemical separations are described in this article. In addition, recent trends in the use of radiochemical separations in neutron activation analysis, in dating applications, in fission product studies and in the study of transactinide elements are briefly described with specific examples. (author). 52 refs

  19. Evaluation of Enzymatically Modified Soy Protein Isolate Film Forming Solution and Film at Different Manufacturing Conditions.

    Science.gov (United States)

    Mohammad Zadeh, Elham; O'Keefe, Sean F; Kim, Young-Teck; Cho, Jin-Hun

    2018-04-01

    The effects of transglutaminase on soy protein isolate (SPI) film forming solution and films were investigated by rheological behavior and physicochemical properties based on different manufacturing conditions (enzyme treatments, enzyme incubation times, and protein denaturation temperatures). Enzymatic crosslinking reaction and changes in molecular weight distribution were confirmed by viscosity measurement and SDS-PAGE, respectively, compared to 2 controls: the nonenzyme treated and the deactivated enzyme treated. Films treated with both the enzyme and the deactivated enzyme showed significant increase in tensile strength (TS), percent elongation (%E), and initial contact angle of films compared to the nonenzyme control film due to the bulk stabilizers in the commercial enzyme. Water absorption property, protein solubility, Fourier transform infrared (FTIR) and X-ray diffraction (XRD) spectroscopy revealed that enzyme treated SPI film matrix in the molecular structure level, resulted in the changes in physicochemical properties. Based on our observation, the enzymatic treatment at appropriate conditions is a practical and feasible way to control the physical properties of protein based biopolymeric film for many different scientific and industrial areas. Enzymes can make bridges selectively among different amino acids in the structure of protein matrix. Therefore, protein network is changed after enzyme treatment. The behavior of biopolymeric materials is dependent on the network structure to be suitable in different applications such as bioplastics applied in food and pharmaceutical products. In the current research, transglutaminase, as an enzyme, applied in soy protein matrix in different types of forms, activated and deactivated, and different preparation conditions to investigate its effects on different properties of the new bioplastic film. © 2018 Institute of Food Technologists®.

  20. Modified expression of several sperm proteins after chronic exposure to the antiandrogenic compound vinclozolin.

    Science.gov (United States)

    Auger, Jacques; Eustache, Florence; Maceiras, Paula; Broussard, Cédric; Chafey, Philippe; Lesaffre, Corinne; Vaiman, Daniel; Camoin, Luc; Auer, Jana

    2010-10-01

    Little is known about the molecular impact of in vivo exposure to endocrine disruptors (EDs) on sperm structures and functions. We recently reported that the lifelong exposure of rats to the antiandrogenic compound vinclozolin results in low epididymal weight, changes in sperm kinematic parameters, and immature sperm chromatin condensation, together with the impairment of several fertility end points. These results led us to focus specifically on possible molecular abnormalities in sperm. Sperm samples were recovered from the frozen epididymides of rats exposed during the previous study. The proteins present in the samples from six exposed and six control rats were analyzed in pairs, by two-dimensional fluorescence difference gel electrophoresis, to investigate possible exposure-induced changes to sperm protein profiles. Twelve proteins, from the 380 matched spots observed in at least five gels, were present in larger or smaller amounts after vinclozolin exposure. These proteins were identified by mass spectrometry, and several are known to play a crucial role in the sperm fertilizing ability, among which, two mitochondrial enzymes, malate dehydrogenase 2 and aldehyde dehydrogenase (both of which were present in smaller amounts after treatment) and A-kinase anchor protein 4 (larger amounts of precursor after treatment). Finally, Ingenuity Pathway Analysis revealed highly significant interactions between proteins over- and underexpressed after treatment. This is the first study to show an association between in vivo exposure to an ED and changes to the sperm protein profile. These modifications may be at least partly responsible for the reproductive abnormalities and impaired fertility recently reported in this rat model of vinclozolin exposure.

  1. Arabidopsis leucine-rich repeat extensin (LRX) proteins modify cell wall composition and influence plant growth.

    Science.gov (United States)

    Draeger, Christian; Ndinyanka Fabrice, Tohnyui; Gineau, Emilie; Mouille, Grégory; Kuhn, Benjamin M; Moller, Isabel; Abdou, Marie-Therese; Frey, Beat; Pauly, Markus; Bacic, Antony; Ringli, Christoph

    2015-06-24

    Leucine-rich repeat extensins (LRXs) are extracellular proteins consisting of an N-terminal leucine-rich repeat (LRR) domain and a C-terminal extensin domain containing the typical features of this class of structural hydroxyproline-rich glycoproteins (HRGPs). The LRR domain is likely to bind an interaction partner, whereas the extensin domain has an anchoring function to insolubilize the protein in the cell wall. Based on the analysis of the root hair-expressed LRX1 and LRX2 of Arabidopsis thaliana, LRX proteins are important for cell wall development. The importance of LRX proteins in non-root hair cells and on the structural changes induced by mutations in LRX genes remains elusive. The LRX gene family of Arabidopsis consists of eleven members, of which LRX3, LRX4, and LRX5 are expressed in aerial organs, such as leaves and stem. The importance of these LRX genes for plant development and particularly cell wall formation was investigated. Synergistic effects of mutations with gradually more severe growth retardation phenotypes in double and triple mutants suggest a similar function of the three genes. Analysis of cell wall composition revealed a number of changes to cell wall polysaccharides in the mutants. LRX3, LRX4, and LRX5, and most likely LRX proteins in general, are important for cell wall development. Due to the complexity of changes in cell wall structures in the lrx mutants, the exact function of LRX proteins remains to be determined. The increasingly strong growth-defect phenotypes in double and triple mutants suggests that the LRX proteins have similar functions and that they are important for proper plant development.

  2. Genetically modified anthrax lethal toxin safely delivers whole HIV protein antigens into the cytosol to induce T cell immunity

    Science.gov (United States)

    Lu, Yichen; Friedman, Rachel; Kushner, Nicholas; Doling, Amy; Thomas, Lawrence; Touzjian, Neal; Starnbach, Michael; Lieberman, Judy

    2000-07-01

    Bacillus anthrax lethal toxin can be engineered to deliver foreign proteins to the cytosol for antigen presentation to CD8 T cells. Vaccination with modified toxins carrying 8-9 amino acid peptide epitopes induces protective immunity in mice. To evaluate whether large protein antigens can be used with this system, recombinant constructs encoding several HIV antigens up to 500 amino acids were produced. These candidate HIV vaccines are safe in animals and induce CD8 T cells in mice. Constructs encoding gag p24 and nef stimulate gag-specific CD4 proliferation and a secondary cytotoxic T lymphocyte response in HIV-infected donor peripheral blood mononuclear cells in vitro. These results lay the foundation for future clinical vaccine studies.

  3. Coxsackievirus protein 2B modifies endoplasmic reticulum membrane and plasma membrane permeability and facilitates virus release.

    Science.gov (United States)

    van Kuppeveld, F J; Hoenderop, J G; Smeets, R L; Willems, P H; Dijkman, H B; Galama, J M; Melchers, W J

    1997-01-01

    Digital-imaging microscopy was performed to study the effect of Coxsackie B3 virus infection on the cytosolic free Ca2+ concentration and the Ca2+ content of the endoplasmic reticulum (ER). During the course of infection a gradual increase in the cytosolic free Ca2+ concentration was observed, due to the influx of extracellular Ca2+. The Ca2+ content of the ER decreased in time with kinetics inversely proportional to those of viral protein synthesis. Individual expression of protein 2B was sufficient to induce the influx of extracellular Ca2+ and to release Ca2+ from ER stores. Analysis of mutant 2B proteins showed that both a cationic amphipathic alpha-helix and a second hydrophobic domain in 2B were required for these activities. Consistent with a presumed ability of protein 2B to increase membrane permeability, viruses carrying a mutant 2B protein exhibited a defect in virus release. We propose that 2B gradually enhances membrane permeability, thereby disrupting the intracellular Ca2+ homeostasis and ultimately causing the membrane lesions that allow release of virus progeny. PMID:9218794

  4. An OGA-Resistant Probe Allows Specific Visualization and Accurate Identification of O-GlcNAc-Modified Proteins in Cells.

    Science.gov (United States)

    Li, Jing; Wang, Jiajia; Wen, Liuqing; Zhu, He; Li, Shanshan; Huang, Kenneth; Jiang, Kuan; Li, Xu; Ma, Cheng; Qu, Jingyao; Parameswaran, Aishwarya; Song, Jing; Zhao, Wei; Wang, Peng George

    2016-11-18

    O-linked β-N-acetyl-glucosamine (O-GlcNAc) is an essential and ubiquitous post-translational modification present in nucleic and cytoplasmic proteins of multicellular eukaryotes. The metabolic chemical probes such as GlcNAc or GalNAc analogues bearing ketone or azide handles, in conjunction with bioorthogonal reactions, provide a powerful approach for detecting and identifying this modification. However, these chemical probes either enter multiple glycosylation pathways or have low labeling efficiency. Therefore, selective and potent probes are needed to assess this modification. We report here the development of a novel probe, 1,3,6-tri-O-acetyl-2-azidoacetamido-2,4-dideoxy-d-glucopyranose (Ac 3 4dGlcNAz), that can be processed by the GalNAc salvage pathway and transferred by O-GlcNAc transferase (OGT) to O-GlcNAc proteins. Due to the absence of a hydroxyl group at C4, this probe is less incorporated into α/β 4-GlcNAc or GalNAc containing glycoconjugates. Furthermore, the O-4dGlcNAz modification was resistant to the hydrolysis of O-GlcNAcase (OGA), which greatly enhanced the efficiency of incorporation for O-GlcNAcylation. Combined with a click reaction, Ac 3 4dGlcNAz allowed the selective visualization of O-GlcNAc in cells and accurate identification of O-GlcNAc-modified proteins with LC-MS/MS. This probe represents a more potent and selective tool in tracking, capturing, and identifying O-GlcNAc-modified proteins in cells and cell lysates.

  5. Electrochemical immunosensors for the detection of survival motor neuron (SMN) protein using different carbon nanomaterials-modified electrodes.

    Science.gov (United States)

    Eissa, Shimaa; Alshehri, Nawal; Rahman, Anas M Abdel; Dasouki, Majed; Abu-Salah, Khalid M; Zourob, Mohammed

    2018-03-15

    Spinal muscular atrophy is an untreatable potentially fatal hereditary disorder caused by loss-of-function mutations in the survival motor neuron (SMN) 1 gene which encodes the SMN protein. Currently, definitive diagnosis relies on the demonstration of biallelic pathogenic variants in SMN1 gene. Therefore, there is an urgent unmet need to accurately quantify SMN protein levels for screening and therapeutic monitoring of symptomatic newborn and SMA patients, respectively. Here, we developed a voltammetric immunosensor for the sensitive detection of SMN protein based on covalently functionalized carbon nanofiber-modified screen printed electrodes. A comparative study of six different carbon nanomaterial-modified electrodes (carbon, graphene (G), graphene oxide (GO), single wall carbon nanotube (SWCNT), multi-wall carbon nanotube (MWCNT), and carbon nanofiber (CNF)) was performed. 4-carboxyphenyl layers were covalently grafted on the six electrodes by electroreduction of diazonium salt. Then, the terminal carboxylic moieties on the electrodes surfaces were utilized to immobilize the SMN antibody via EDC/NHS chemistry and to fabricate the immunosensors. The electrochemical characterization and analytical performance of the six immunosensors suggest that carbon nanofiber is a better electrode material for the SMN immunosensor. The voltammetric SMN carbon nanofiber-based immunosensor showed high sensitivity (detection limit of 0.75pg/ml) and selectivity against other proteins such as cystic fibrosis transmembrane conductance regulator (CFTR) and dystrophin (DMD). We suggest that this novel biosensor is superior to other developed assays for SMN detection in terms of lower cost, higher sensitivity, simplicity and capability of high throughput screening. Copyright © 2017 Elsevier B.V. All rights reserved.

  6. Self-decomposition of radiochemicals. Principles, control, observations and effects

    International Nuclear Information System (INIS)

    Evans, E.A.

    1976-01-01

    The aim of the booklet is to remind the established user of radiochemicals of the problems of self-decomposition and to inform those investigators who are new to the applications of radiotracers. The section headings are: introduction; radionuclides; mechanisms of decomposition; effects of temperature; control of decomposition; observations of self-decomposition (sections for compounds labelled with (a) carbon-14, (b) tritium, (c) phosphorus-32, (d) sulphur-35, (e) gamma- or X-ray emitting radionuclides, decomposition of labelled macromolecules); effects of impurities in radiotracer investigations; stability of labelled compounds during radiotracer studies. (U.K.)

  7. Mercury determination in geological samples using radiochemical separation

    International Nuclear Information System (INIS)

    Goncalves, Cristina; Favaro, Deborah I.T.

    1997-01-01

    In this work, a radiochemical procedure is presented to increase the neutron activation analysis sensitivity. After irradiation, geological reference materials - Buffalo River Sediment (BRS- - NIST SRM 2704), Lake Sediment (BCR - CRM 280) and GXR-5 (USGS - AEG) - were leached with aqua regia in a Parr bomb placed in a domestic microwave oven and then bismuth diethyl dithiocarbamate was used to pre concentrate mercury by solvent extraction. This procedure eliminates the interference from 279 keV Se-75 photopeak and background radiation from 511 keV Cu-64 photopeak. (author). 15 refs., 2 figs., 1 tab

  8. International intercalibration as a method for control of radiochemical analyses

    International Nuclear Information System (INIS)

    Angelova, A.; Totseva, R.; Karaivanova, R.; Dandulova, Z.; Botsova, L.

    1994-01-01

    The participation of the Radioecology Section at the National Centre for Radiology and Radiation Protection (NCRRP) in the International Interlaboratory Comparison of radiochemical analyses organized by WHO is reported. The method of evaluating accuracy of the results from inter calibrations concerning radionuclide determination of environmental samples is outlined. The data from analysis of cesium 137, strontium 90 and radium 226 in milk, sediments, soil and seaweed made by 21 laboratories are presented. They show a good accuracy values of the results from NCRRP. 1 tab. 2 figs., 6 refs

  9. Low temperature radio-chemical energy conversion processes

    International Nuclear Information System (INIS)

    Gomberg, H.J.

    1986-01-01

    This patent describes a radio-chemical method of converting radiated energy into chemical energy form comprising the steps of: (a) establishing a starting chemical compound in the liquid phase that chemically reacts endothermically to radiation and heat energy to produce a gaseous and a solid constituent of the compound, (b) irradiating the compound in its liquid phase free of solvents to chemically release therefrom in response to the radiation the gaseous and solid constituents, (c) physically separating the solid and gaseous phase constituents from the liquid, and (d) chemically processing the constituents to recover therefrom energy stored therein by the irradiation step (b)

  10. Rapid, radiochemical-ligand binding assay for methotrexate

    International Nuclear Information System (INIS)

    Caston, J.D.

    1976-01-01

    A radiochemical ligand binding assay for methotrexate is provided. A binder factor comprising a partially purified dihydrofolic acid reductase preparation is employed. The binder factor is conveniently prepared by homogenizing a factor containing animal organ such as liver, and extracting with isotonic saline and ammonium sulfate. A binder cofactor, NADPH 2 , is also employed in the binding reaction. The procedure contemplates both direct and sequential assay techniques, and it is not interfered with by vast excesses of many natural folate derivatives. 12 claims, 6 drawing figures

  11. Radiochemical schemes of obtaining 89Sr and 90Y radionuclides

    International Nuclear Information System (INIS)

    Usarov, Z. O.

    2010-03-01

    Key words: strontium-89, yttrium-90, extraction and extraction-chromatographic purification of radionuclides, radiopharmaceuticals. Subjects of research: strontium-89 and yttrium-90 radionuclides and their chloride forms. Purpose of work is developing of radiochemical technologies on obtaining of 89 Sr and 90 Y on the WWR-SM reactor with high radionuclide purity. Methods of research: extraction and extraction-chromatographic methods of radionuclides separation, beta- and gamma-spectrometric methods of activity measuring. The results obtained and their novelty: Were determined the conformity to laws of Y and Sr distribution in two-phase systems TBP-HNO 3 , TBP-NH 4 NO 3 , TBP-HCI, HDEHP-NO 3 , HDEHP-NH 4 NO 3 and HDEHP-HCI. Were determined the conformity to laws of Y and Sr distribution in systems with craun ethers DB-18K-6 and DTBDB-18K-6 from water solutions of HNO 3 . Radiochemical technologies on obtaining of 89 Sr and 90 Y radionuclides including radiochemical process of yttrium target with using the systems TBP-HNO 3 and HDEHP/Teflone were developed. Practical value: the radiochemical technology of obtaining 89 Sr with high radionuclide purity was developed. The method of preparation a chloride compound of 89 SrCl 2 which is used as a drug form for preparation of 89 Sr- 'Metastron' was developed. The relatively simple method of on the way obtaining 90 Y in the reactor with high radionuclidic purity that is useful for follow using in medical practice was offered. Degree of embed and economic effectivity: the developed technologies have approbation in manufacturing conditions in Radiopreparat Enterprise of INP AS RU and were offered for receiving of domestic preparations against of import foreign analogues. The statement about using the invention by obtained patent is attached to dissertation. Field of application: the received results will be introduced in manufacture at Radiopreparat Enterprise of INP AS RU for receiving of domestic preparations

  12. Guiding Principles for Sustainable Existing Buildings: Radiochemical Processing Laboratory

    Energy Technology Data Exchange (ETDEWEB)

    Pope, Jason E. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States)

    2013-11-11

    In 2006, the United States (U.S.) Department of Energy (DOE) signed the Federal Leadership in High Performance and Sustainable Buildings Memorandum of Understanding (MOU), along with 21 other agencies. Pacific Northwest National Laboratory (PNNL) is exceeding this requirement and, currently, about 25 percent of its buildings are High Performance and Sustainable Buildings. The pages that follow document the Guiding Principles conformance effort for the Radiochemical Processing Laboratory (RPL) at PNNL. The RPL effort is part of continued progress toward a building inventory that is 100 percent compliant with the Guiding Principles.

  13. Preparation of proton rich radionuclides in support of radiochemical analysis

    International Nuclear Information System (INIS)

    Jerome, Simon; Larijani, Cyrus; Parker, David

    2012-01-01

    The production of proton rich radionuclides supports a wide range of radiochemical analyses via radioactive yield tracers ( 95m Tc and 236 Pu). In recent years, NPL and the University of Birmingham cyclotron have collaborated to produce these, and other, radionuclides. - Highlights: ► In this paper we options for the production of Tc and Pu tracers. ► The irradiation and measurement of targets producing Tc-95 m and Pu-236 are described. ► Options for production are discussed. ► The results of this study and future work needed are described.

  14. Studies on some Indian paints for radiochemical plants

    International Nuclear Information System (INIS)

    Mahesh Kumar, V.V.; Srinivasan, R.; Natarajan, R.

    1996-01-01

    The choice of paints in areas subjected to contamination and radiation in nuclear installation need special attention. The types of generic coatings are examined with reference to these requirements. Among those examined, certain types of epoxy paints are found to be attractive for these applications. Samples of epoxy paints obtained from some Indian manufacturers are tested for their suitability. Decontaminability and radiation resistance properties are also evaluated with special reference to radiochemical plants. Important specifications for such applications are listed. This report summarizes the results of these studies. (author)

  15. N(epsilon)-carboxymethyllysine-modified proteins are unable to bind to RAGE and activate an inflammatory response.

    Science.gov (United States)

    Buetler, Timo M; Leclerc, Estelle; Baumeyer, Alexandra; Latado, Helia; Newell, John; Adolfsson, Oskar; Parisod, Véronique; Richoz, Janique; Maurer, Sarah; Foata, Francis; Piguet, Dominique; Junod, Sylviane; Heizmann, Claus W; Delatour, Thierry

    2008-03-01

    Advanced glycation endproducts (AGEs) containing carboxymethyllysine (CML) modifications are generally thought to be ligands of the receptor for AGEs, RAGEs. It has been argued that this results in the activation of pro-inflammatory pathways and diseases. However, it has not been shown conclusively that a CML-modified protein can interact directly with RAGE. Here, we have analyzed whether beta-lactoglobulin (bLG) or human serum albumin (HSA) modified chemically to contain only CML (10-40% lysine modification) can (i) interact with RAGE in vitro and (ii) interact with and activate RAGE in lung epithelial cells. Our results show that CML-modified bLG or HSA are unable to bind to RAGE in a cell-free assay system (Biacore). Furthermore, they are unable to activate pro-inflammatory signaling in the cellular system. Thus, CML probably does not form the necessary structure(s) to interact with RAGE and activate an inflammatory signaling cascade in RAGE-expressing cells.

  16. Pokemon siRNA Delivery Mediated by RGD-Modified HBV Core Protein Suppressed the Growth of Hepatocellular Carcinoma.

    Science.gov (United States)

    Kong, Jing; Liu, Xiaoping; Jia, Jianbo; Wu, Jinsheng; Wu, Ning; Chen, Jun; Fang, Fang

    2015-10-01

    Hepatocellular carcinoma (HCC) is a deadly human malignant tumor that is among the most common cancers in the world, especially in Asia. Hepatitis B virus (HBV) infection has been well established as a high risk factor for hepatic malignance. Studies have shown that Pokemon is a master oncogene for HCC growth, suggesting it as an ideal therapeutic target. However, efficient delivery system is still lacking for Pokemon targeting treatment. In this study, we used core proteins of HBV, which is modified with RGD peptides, to construct a biomimetic vector for the delivery of Pokemon siRNAs (namely, RGD-HBc-Pokemon siRNA). Quantitative PCR and Western blot assays revealed that RGD-HBc-Pokemon siRNA possessed the highest efficiency of Pokemon suppression in HCC cells. In vitro experiments further indicated that RGD-HBc-Pokemon-siRNA exerted a higher tumor suppressor activity on HCC cell lines, evidenced by reduced proliferation and attenuated invasiveness, than Pokemon-siRNA or RGD-HBc alone. Finally, animal studies demonstrated that RGD-HBc-Pokemon siRNA suppressed the growth of HCC xenografts in mice by a greater extent than Pokemon-siRNA or RGD-HBc alone. Based on the above results, Pokemon siRNA delivery mediated by RGD-modified HBV core protein was shown to be an effective strategy of HCC gene therapy.

  17. Re-partitioning of Cu and Zn isotopes by modified protein expression

    Directory of Open Access Journals (Sweden)

    Ragnarsdottir K Vala

    2008-10-01

    Full Text Available Abstract Cu and Zn have naturally occurring non radioactive isotopes, and their isotopic systematics in a biological context are poorly understood. In this study we used double focussing mass spectroscopy to determine the ratios for these isotopes for the first time in mouse brain. The Cu and Zn isotope ratios for four strains of wild-type mice showed no significant difference (δ65Cu -0.12 to -0.78 permil; δ66Zn -0.23 to -0.48 permil. We also looked at how altering the expression of a single copper binding protein, the prion protein (PrP, alters the isotope ratios. Both knockout and overexpression of PrP had no significant effect on the ratio of Cu isotopes. Mice brains expressing mutant PrP lacking the known metal binding domain have δ65Cu isotope values of on average 0.57 permil higher than wild-type mouse brains. This implies that loss of the copper binding domain of PrP increases the level of 65Cu in the brain. δ66Zn isotope values of the transgenic mouse brains are enriched for 66Zn to the wild-type mouse brains. Here we show for the first time that the expression of a single protein can alter the partitioning of metal isotopes in mouse brains. The results imply that the expression of the prion protein can alter cellular Cu isotope content.

  18. Mining the secretome of root-knot nematodes for cell wall modifying proteins

    NARCIS (Netherlands)

    Roze, E.H.A.

    2008-01-01

    The products of parasitism genes in nematodes must be secreted to reach their targets at the nematode-plant interface. These nematode secretory proteins are therefore recognised to play an important role in the nematode-plant interaction and as a result have been subject of intense study for years.

  19. Facility Effluent Monitoring Plan for the 325 Radiochemical Processing Laboratory

    International Nuclear Information System (INIS)

    Shields, K.D.; Ballinger, M.Y.

    1999-03-01

    This Facility Effluent Monitoring Plan (FEMP) has been prepared for the 325 Building Radiochemical Processing Laboratory (RPL) at the Pacific Northwest National Laboratory (PNNL) to meet the requirements in DOE Order 5400.1, ''General Environmental Protection Programs.'' This FEMP has been prepared for the RPL primarily because it has a ''major'' (potential to emit >0.1 mrem/yr) emission point for radionuclide air emissions according to the annual National Emission Standards for Hazardous Air Pollutants (NESHAP) assessment performed. This section summarizes the airborne and liquid effluents and the inventory based NESHAP assessment for the facility. The complete monitoring plan includes characterization of effluent streams, monitoring/sampling design criteria, a description of the monitoring systems and sample analysis, and quality assurance requirements. The RPL at PNNL houses radiochemistry research, radioanalytical service, radiochemical process development, and hazardous and radioactive mixed waste treatment activities. The laboratories and specialized facilities enable work ranging from that with nonradioactive materials to work with picogram to kilogram quantities of fissionable materials and up to megacurie quantities of other radionuclides. The special facilities within the building include two shielded hot-cell areas that provide for process development or analytical chemistry work with highly radioactive materials and a waste treatment facility for processing hazardous, mixed radioactive, low-level radioactive, and transuranic wastes generated by PNNL activities

  20. A radiochemical analyses of metastudtite and leachates from spent fuel

    International Nuclear Information System (INIS)

    McNamara, Bruce K.; Hanson, Brady D.; Buck, Edgar C.; Soderquist, Chuck Z.

    2004-01-01

    Immersion of commercial spent nuclear fuel (CSNF) in deionized water produced two novel corrosion products after a two-year contact period. Another unexpected result was that suspensions of aggregates were observed to form at the air-water interface for each of five samples. These solids were characterized, by SEM and XRD to be nearly pure metastudtite (UO4-2H2O); while the corrosion present on the surface of the fuel itself was determined to be studtite (UO4-2H2O). The occurrence of the floating phase prompted a radiochemical analysis of these solids. This chemical analysis was a unique opportunity to study the relatively pure corrosion phase for incorporation of radionuclides. The analysis indicated that high concentration of 90Sr, 137Cs, 99Tc, and that lower concentrations 237Np, 238, 239Pu and 243, 244Cm had partitioned with the air-water interface aggregates. The concentrations of 241Am were two orders of magnitude lower than the expected inventory in the suspended solids. The radiochemical analyses of the several leachate samples provide preliminary solubility data for the hydrogen peroxide leaching of CSNF and these data are compared to leaching of the same fuel in J-13 and deionized waters. The extent of fuel dissolution in these media are discussed

  1. Facility Effluent Monitoring Plan for the 325 Radiochemical Processing Laboratory

    Energy Technology Data Exchange (ETDEWEB)

    Shields, K.D.; Ballinger, M.Y.

    1999-04-02

    This Facility Effluent Monitoring Plan (FEMP) has been prepared for the 325 Building Radiochemical Processing Laboratory (RPL) at the Pacific Northwest National Laboratory (PNNL) to meet the requirements in DOE Order 5400.1, ''General Environmental Protection Programs.'' This FEMP has been prepared for the RPL primarily because it has a ''major'' (potential to emit >0.1 mrem/yr) emission point for radionuclide air emissions according to the annual National Emission Standards for Hazardous Air Pollutants (NESHAP) assessment performed. This section summarizes the airborne and liquid effluents and the inventory based NESHAP assessment for the facility. The complete monitoring plan includes characterization of effluent streams, monitoring/sampling design criteria, a description of the monitoring systems and sample analysis, and quality assurance requirements. The RPL at PNNL houses radiochemistry research, radioanalytical service, radiochemical process development, and hazardous and radioactive mixed waste treatment activities. The laboratories and specialized facilities enable work ranging from that with nonradioactive materials to work with picogram to kilogram quantities of fissionable materials and up to megacurie quantities of other radionuclides. The special facilities within the building include two shielded hot-cell areas that provide for process development or analytical chemistry work with highly radioactive materials and a waste treatment facility for processing hazardous, mixed radioactive, low-level radioactive, and transuranic wastes generated by PNNL activities.

  2. Identification of liver protein targets modified by tienilic acid metabolites using a two-dimensional Western blot-mass spectrometry approach

    Science.gov (United States)

    Methogo, Ruth Menque; Dansette, Patrick M.; Klarskov, Klaus

    2007-12-01

    A combined approach based on two-dimensional electrophoresis-immuno-blotting and nanoliquid chromatography coupled on-line with electrospray ionization mass spectrometry (nLC-MS/MS) was used to identify proteins modified by a reactive intermediate of tienilic acid (TA). Liver homogenates from rats exposed to TA were fractionated using ultra centrifugation; four fractions were obtained and subjected to 2D electrophoresis. Following transfer to PVDF membranes, modified proteins were visualized after India ink staining, using an anti-serum raised against TA and ECL detection. Immuno-reactive spots were localized on the PVDF membrane by superposition of the ECL image, protein spots of interest were excised, digested on the membrane with trypsin followed by nLC-MS/MS analysis and protein identification. A total of 15 proteins were identified as likely targets modified by a TA reactive metabolite. These include selenium binding protein 2, senescence marker protein SMP-30, adenosine kinase, Acy1 protein, adenosylhomocysteinase, capping protein (actin filament), protein disulfide isomerase, fumarylacetoacetase, arginase chain A, ketohexokinase, proteasome endopeptidase complex, triosephosphate isomerase, superoxide dismutase, dna-type molecular chaperone hsc73 and malate dehydrogenase.

  3. Incorporation of chimeric HIV-SIV-Env and modified HIV-Env proteins into HIV pseudovirions

    International Nuclear Information System (INIS)

    Devitt, Gerard; Emerson, Vanessa; Holtkotte, Denise; Pfeiffer, Tanya; Pisch, Thorsten; Bosch, Valerie

    2007-01-01

    Low level incorporation of the viral glycoprotein (Env) into human immunodeficiency virus (HIV) particles is a major drawback for vaccine strategies against HIV/AIDS in which HIV particles are used as immunogen. Within this study, we have examined two strategies aimed at achieving higher levels of Env incorporation into non-infectious pseudovirions (PVs). First, we have generated chimeric HIV/SIV Env proteins containing the truncated C-terminal tail region of simian immunodeficiency virus (SIV)mac239-Env767 stop , which mediates strongly increased incorporation of SIV-Env into SIV particles. In a second strategy, we have employed a truncated HIV-Env protein (Env-Tr752 N750K ) which we have previously demonstrated to be incorporated into HIV virions, generated in infected T-cells, to a higher level than that of Wt-HIV-Env. Although the chimeric HIV/SIV Env proteins were expressed at the cell surface and induced increased levels of cell-cell fusion in comparison to Wt-HIV-Env, they did not exhibit increased incorporation into either HIV-PVs or SIV-PVs. Only Env-Tr752 N750K exhibited significantly higher (threefold) levels of incorporation into HIV-PVs, an improvement, which, although not dramatic, is worthwhile for the large-scale preparation of non-infectious PVs for vaccine studies aimed at inducing Env humoral responses

  4. Immunogenicity of virus-like particles containing modified goose parvovirus VP2 protein.

    Science.gov (United States)

    Chen, Zongyan; Li, Chuanfeng; Zhu, Yingqi; Wang, Binbin; Meng, Chunchun; Liu, Guangqing

    2012-10-01

    The major capsid protein VP2 of goose parvovirus (GPV) expressed using a baculovirus expression system (BES) assembles into virus-like particles (VLPs). To optimize VP2 gene expression in Sf9 cells, we converted wild-type VP2 (VP2) codons into codons that are more common in insect genes. This change greatly increased VP2 protein production in Sf9 cells. The protein generated from the codon-optimized VP2 (optVP2) was detected by immunoblotting and an indirect immunofluorescence assay (IFA). Transmission electron microscopy analysis revealed the formation of VLPs. These findings indicate that optVP2 yielded stable and high-quality VLPs. Immunogenicity assays revealed that the VLPs are highly immunogenic, elicit a high level of neutralizing antibodies and provide protection against lethal challenge. The antibody levels appeared to be directly related to the number of GP-Ag-positive hepatocytes. The variation trends for GP-Ag-positive hepatocytes were similar in the vaccine groups. In comparison with the control group, the optVP2 VLPs groups exhibited obviously better responses. These data indicate that the VLPs retained immunoreactivity and had strong immunogenicity in susceptible geese. Thus, GPV optVP2 appears to be a good candidate for the vaccination of goslings. Copyright © 2012 Elsevier B.V. All rights reserved.

  5. Interaction Of Calcium Phosphate Nanoparticles With Human Chorionic Gonadotropin Modifies Secondary And Tertiary Protein Structure

    Directory of Open Access Journals (Sweden)

    Al-Hakeim Hussein K

    2015-12-01

    Full Text Available Calcium phosphate nanoparticles (CaPNP have good biocompatibility and bioactivity inside human body. In this study, the interaction between CaPNP and human chorionic gonadotropin (hCG was analyzed to determine the changes in the protein structure in the presence of CaPNP and the quantity of protein adsorbed on the CaPNP surface. The results showed a significant adsorption of hCG on the CaPNP nanoparticle surface. The optimal fit was achieved using the Sips isotherm equation with a maximum adsorption capacity of 68.23 µg/mg. The thermodynamic parameters, including ∆H° and ∆G°, of the adsorption process are positive, whereas ∆S° is negative. The circular dichroism results of the adsorption of hCG on CaPNP showed the changes in its secondary structure; such changes include the decomposition of α-helix strand and the increase in β-pleated sheet and random coil percentages. Fluorescence study indicated minimal changes in the tertiary structure near the microenvironment of the aromatic amino acids such as tyrosine and phenyl alanine caused by the interaction forces between the CaPNP and hCG protein. The desorption process showed that the quantity of the hCG desorbed significantly increases as temperature increases, which indicates the weak forces between hCG and the surface.

  6. Instant thin layer chromatography (ITLC) - the most true and convenient method for the determination of radiochemical purity of Tc-99m Labeled radiopharmaceuticals

    International Nuclear Information System (INIS)

    Anwar-Ul-Azim, M.; Ansari, M.I.H.

    2007-01-01

    Full text: Determination of radiochemical purity usually involves paper chromatography method or some column chromatography such as ion exchange or gel filtration technique. In this study, a modified chromatography method with thin-layer support called instant thin Layer chromatography (ITLC) was used for the measurement of radiochemical purity. The aim of the study was to present instant thin layer chromatography-Silica-Gel (ITLC-SG) technique as a fairly rapid , convenient and inexpensive system for the screening of radiochemical impurities of 99mTc-labeled radiopharmaceuticals resulting from physical , chemical and/or radiation decomposition. Materials and Methods: The experiment was carried out at the Institute of Nuclear Medicine and Ultrasound, Dhaka for a period of 10 months from August '05 to May '06. Radiochemical purity of seven types of 99mTc-labeled radiopharmaceuticals, namely 99mTc-DTPA,99mTc- MI I, 99m Tc - HDP, 99m Tc - DMSA, 99m Tc - Tetofosmin, 99mTc-Fyton and 99mTc-ECD were measured by the use of two solvent dual-instant thin layer chromatography systems. Commercially available instant thin layer chromatography plate ( 20cm x20cm) impregnated with silica gel (ITLC-SG: Merck, Germany) was used as a stationary phase for the measurement of radiochemical impurities.Acetone, Ethyl methyl ketone, chloroform and methanol were used to measure the amount of free pertechnetate in the labelled radiopharmaceuticals and 0.9% saline was used in all cases as solvent (Mobile phase) to measure the amount of the hydrolyzed-reduced technetium colloid. Results: The results showed that the measured radiochemical purity of 99mTc-DTPA, 99mTc-MIBI, 99mTc-HDP, 99mTc-DMSA, 99mTc-Tetofosmin, 99mTc-Fyton and 99mTc-ECD were acceptable for all the studied samples except two samples of 99mTc-MIBI, one sample of 99mTc-Fyton and two samples of 99mTc-ECD, where the measured radiochemical purity were not within the acceptable RCP limits of commonly used radiopharmaceuticals (RCP

  7. Receptor activity modifying proteins (RAMPs) interact with the VPAC1 receptor: evidence for differential RAMP modulation of multiple signalling pathways

    International Nuclear Information System (INIS)

    Christopoulos, G.; Morfis, M.; Sexton, P.M.; Christopoulos, A.; Laburthe, M.; Couvineau, A.

    2001-01-01

    Full text: Receptor activity modifying proteins (RAMP) constitute a family of three accessory proteins that affect the expression and/or phenotype of the calcitonin receptor (CTR) or CTR-like receptor (CRLR). In this study we screened a range of class II G protein-coupled receptors (PTH1, PTH2, GHRH, VPAC1, VPAC2 receptors) for possible RAMP interactions by measurement of receptor-induced translocation of c-myc tagged RAMP1 or HA tagged RAMP3. Of these, only the VPAC1 receptor caused significant translocation of c-myc-RAMP1 or HA-RAMP3 to the cell surface. Co-transfection of VPAC1 and RAMPs did not alter 125 I-VIP binding and specificity. VPAC1 receptor function was subsequently analyzed through parallel determinations of cAMP accumulation and phosphoinositide (PI) hydrolysis in the presence and absence of each of the three RAMPs. In contrast to CTR-RAMP interaction, where there was an increase in cAMP Pharmacologisand a decrease in PI hydrolysis, VPAC1-RAMP interaction was characterized by a specific increase in agonist-mediated PI hydrolysis when co-transfected with RAMP2. This change was due to an enhancement of Emax with no change in EC 50 value for VIP. No significant change in cAMP accumulation was observed. This is the first demonstration of an interaction of RAMPs with a G protein-coupled receptor outside the CTR family and may suggest a more generalized role for RAMPs in modulating G protein-coupled receptor signaling. Copyright (2001) Australasian Society of Clinical and Experimental Pharmacologists and Toxicologists

  8. High-protein diet differently modifies intestinal goblet cell characteristics and mucosal cytokine expression in ileum and colon.

    Science.gov (United States)

    Lan, Annaïg; Andriamihaja, Mireille; Blouin, Jean-Marc; Liu, Xinxin; Descatoire, Véronique; Desclée de Maredsous, Caroline; Davila, Anne-Marie; Walker, Francine; Tomé, Daniel; Blachier, François

    2015-01-01

    We have previously shown that high-protein (HP) diet ingestion causes marked changes in the luminal environment of the colonic epithelium. This study aimed to evaluate the impact of such modifications on small intestinal and colonic mucosa, two segments with different transit time and physiological functions. Rats were fed with either normal protein (NP; 14% protein) or HP (53% protein) isocaloric diet for 2 weeks, and parameters related to intestinal mucous-secreting cells and to several innate/adaptive immune characteristics (myeloperoxidase activity, cytokine and epithelial TLR expression, proportion of immune cells in gut-associated lymphoid tissues) were measured in the ileum and colon. In ileum from HP animals, we observed hyperplasia of mucus-producing cells concomitant with an increased expression of Muc2 at both gene and protein levels, reduction of mucosal myeloperoxidase activity, down-regulation of Tlr4 gene expression in enterocytes and down-regulation of mucosal Th cytokines associated with CD4+ lymphocyte reduction in mesenteric lymph nodes. These changes coincided with an increased amount of acetate in the ileal luminal content. In colon, HP diet ingestion resulted in a lower number of goblet cells at the epithelial surface but increased goblet cell number in colonic crypts together with an increased Muc3 and a slight reduction of Il-6 gene expression. Our data suggest that HP diet modifies the goblet cell distribution in colon and, in ileum, increases goblet cell activity and decreases parameters related to basal gut inflammatory status. The impact of HP diet on intestinal mucosa in terms of beneficial or deleterious effects is discussed. Copyright © 2015 Elsevier Inc. All rights reserved.

  9. Development of radiochemical method of analysis of binding of tritium labeled drotaverine hydrochloride with human blood serum albumin

    International Nuclear Information System (INIS)

    Kim, A.A.; Djuraeva, G.T.; Shukurov, B.V.; Mavlyanov, I.R.

    2004-01-01

    Full text: The albumin, being a basic functional linkage of numerous endogenous and exogenous substances is the most important protein of blood plasma. At the diseases connected to liver disfunction, collected in blood metabolite reduce connecting ability of albumino. The aim of the present research was a development of radiochemical method of determination of ability of albumin to bind the tritium labeled preparation drotaverine hydrochloride (no - spa). We had developed a micromethod of definition of connecting ability of albumin, allowing to analyse 20 mkl of blood serum. The method consists in incubation of tritium labeled drotaverine hydrochloride with blood serum in vitro, the following fractionation of serum proteins by gel - filtration on a microcolumn with Sephadex G-25, and direct measurement of the radioactivity connected to fraction of proteins of blood serum. The method has been tested on a series of blood serum of control group of healthy people and on a series of blood serum of patients with hepatitis B. We received quantitative characteristics of binding of drotaverine hydrochloride with albumin of patients with hepatitis B. It was preliminary established that binding ability of serum albumin of children with various forms of acute virus hepatitis tends to decrease in comparison with group of the control. Advantage of the developed radiochemical method is high precision and the high sensitivity of detection of infringement of binding ability of albumin. Application of tritium labeled drotaverine hydrochloride allows to measure directly levels of binding of a preparation with albumin

  10. Modified C-reactive protein is expressed by stroke neovessels and is a potent activator of angiogenesis in vitro.

    Science.gov (United States)

    Slevin, Mark; Matou-Nasri, Sabine; Turu, Marta; Luque, Ana; Rovira, Norma; Badimon, Lina; Boluda, Susana; Potempa, Lawrence; Sanfeliu, Coral; de Vera, Nuria; Krupinski, Jerzy

    2010-01-01

    Native C-reactive protein (nCRP) is a pentameric oligo-protein and an acute phase reactant whose serum expression is increased in patients with inflammatory disease. We have identified by immunohistochemistry, significant expression of a tissue-binding insoluble modified version or monomeric form of CRP (mCRP) associated with angiogenic microvessels in peri-infarcted regions of patients studied with acute ischaemic stroke. mCRP, but not nCRP was expressed in the cytoplasm and nucleus of damaged neurons. mCRP co-localized with CD105, a marker of angiogenesis in regions of revascularisation. In vitro investigations demonstrated that mCRP was preferentially expressed in human brain microvessel endothelial cells following oxygen-glucose deprivation and mCRP (but not column purified nCRP) associated with the endothelial cell surface, and was angiogenic to vascular endothelial cells, stimulating migration and tube formation in matrigel more strongly than fibroblast growth factor-2. The mechanism of signal transduction was not through the CD16 receptor. Western blotting showed that mCRP stimulated phosphorylation of the key down-stream mitogenic signalling protein ERK1/2. Pharmacological inhibition of ERK1/2 phosphorylation blocked the angiogenic effects of mCRP. We propose that mCRP may contribute to the neovascularization process and because of its abundant presence, be important in modulating angiogenesis in both acute stroke and later during neuro-recovery.

  11. Improved innate and adaptive immunostimulation by genetically modified HIV-1 protein expressing NYVAC vectors.

    Directory of Open Access Journals (Sweden)

    Esther D Quakkelaar

    Full Text Available Attenuated poxviruses are safe and capable of expressing foreign antigens. Poxviruses are applied in veterinary vaccination and explored as candidate vaccines for humans. However, poxviruses express multiple genes encoding proteins that interfere with components of the innate and adaptive immune response. This manuscript describes two strategies aimed to improve the immunogenicity of the highly attenuated, host-range restricted poxvirus NYVAC: deletion of the viral gene encoding type-I interferon-binding protein and development of attenuated replication-competent NYVAC. We evaluated these newly generated NYVAC mutants, encoding HIV-1 env, gag, pol and nef, for their ability to stimulate HIV-specific CD8 T-cell responses in vitro from blood mononuclear cells of HIV-infected subjects. The new vectors were evaluated and compared to the parental NYVAC vector in dendritic cells (DCs, RNA expression arrays, HIV gag expression and cross-presentation assays in vitro. Deletion of type-I interferon-binding protein enhanced expression of interferon and interferon-induced genes in DCs, and increased maturation of infected DCs. Restoration of replication competence induced activation of pathways involving antigen processing and presentation. Also, replication-competent NYVAC showed increased Gag expression in infected cells, permitting enhanced cross-presentation to HIV-specific CD8 T cells and proliferation of HIV-specific memory CD8 T-cells in vitro. The recombinant NYVAC combining both modifications induced interferon-induced genes and genes involved in antigen processing and presentation, as well as increased Gag expression. This combined replication-competent NYVAC is a promising candidate for the next generation of HIV vaccines.

  12. Cancer associated aberrant protein o-glycosylation can modify antigen processing and immune response

    DEFF Research Database (Denmark)

    Madsen, Caroline B; Petersen, Cecilie; Lavrsen, Kirstine

    2012-01-01

    Aberrant glycosylation of mucins and other extracellular proteins is an important event in carcinogenesis and the resulting cancer associated glycans have been suggested as targets in cancer immunotherapy. We assessed the role of O-linked GalNAc glycosylation on antigen uptake, processing......, and presentation on MHC class I and II molecules. The effect of GalNAc O-glycosylation was monitored with a model system based on ovalbumin (OVA)-MUC1 fusion peptides (+/- glycosylation) loaded onto dendritic cells co-cultured with IL-2 secreting OVA peptide-specific T cell hybridomas. To evaluate the in vivo...

  13. EMILIN2 (Elastin microfibril interface located protein, potential modifier of thrombosis

    Directory of Open Access Journals (Sweden)

    Hoover-Plow Jane L

    2011-05-01

    Full Text Available Abstract Background Elastin microfibril interface located protein 2 (EMILIN2 is an extracellular glycoprotein associated with cardiovascular development. While other EMILIN proteins are reported to play a role in elastogenesis and coagulation, little is known about EMILIN2 function in the cardiovascular system. The objective of this study was to determine whether EMILIN2 could play a role in thrombosis. Results EMILIN2 mRNA was expressed in 8 wk old C57BL/6J mice in lung, heart, aorta and bone marrow, with the highest expression in bone marrow. In mouse cells, EMILIN2 mRNA expression in macrophages was higher than expression in endothelial cells and fibroblasts. EMILIN2 was identified with cells and extracellular matrix by immunohistochemistry in the carotid and aorta. After carotid ferric chloride injury, EMILIN2 was abundantly expressed in the thrombus and inhibition of EMILIN2 increased platelet de-aggregation after ADP-stimulated platelet aggregation. Conclusions These results suggest EMILIN2 could play a role in thrombosis as a constituent of the vessel wall and/or a component of the thrombus.

  14. A modified GFP facilitates counting membrane protein subunits by step-wise photobleaching in Arabidopsis.

    Science.gov (United States)

    Song, Kai; Xue, Yiqun; Wang, Xiaohua; Wan, Yinglang; Deng, Xin; Lin, Jinxing

    2017-06-01

    Membrane proteins exert functions by forming oligomers or molecular complexes. Currently, step-wise photobleaching has been applied to count the fluorescently labelled subunits in plant cells, for which an accurate and reliable control is required to distinguish individual subunits and define the basal fluorescence. However, the common procedure using immobilized GFP molecules is obviously not applicable for analysis in living plant cells. Using the spatial intensity distribution analysis (SpIDA), we found that the A206K mutation reduced the dimerization of GFP molecules. Further ectopic expression of Myristoyl-GFP A206K driven by the endogenous AtCLC2 promoter allowed imaging of individual molecules at a low expression level. As a result, the percentage of dimers in the transgenic pCLC2::Myristoyl-mGFP A206K line was significantly reduced in comparison to that of the pCLC2::Myristoyl-GFP line, confirming its application in defining the basal fluorescence intensity of GFP. Taken together, our results demonstrated that pCLC2::Myristoyl-mGFP A206K can be used as a standard control for monomer GFP, facilitating the analysis of the step-wise photobleaching of membrane proteins in Arabidopsis thaliana. Copyright © 2017 Elsevier GmbH. All rights reserved.

  15. The analytical of radiochemical purity of tumor receptor imaging agent 99Tcm-octreotide

    International Nuclear Information System (INIS)

    Wang Xufu; Zuo Shuyao; Shao Wenbo; Wang Guoming; Sun Jianwen; Zhang Qin

    2003-01-01

    The radiochemical purity of tumor receptor imaging agent 99 Tc m -octreotide is measured by High Pressure Liquid Chromatography (HPLC) and two systems of chromatography combining method of silver stain. The results show that the radiochemical purity of 98 Tc m -octreotide measured by both methods are effective and correct. It can separate 99 Tc m -octreotide from other radioactive compositions correctly and effectively

  16. Serotonergic Chemosensory Neurons Modify the C. elegans Immune Response by Regulating G-Protein Signaling in Epithelial Cells

    Science.gov (United States)

    Anderson, Alexandra; Laurenson-Schafer, Henry; Partridge, Frederick A.; Hodgkin, Jonathan; McMullan, Rachel

    2013-01-01

    The nervous and immune systems influence each other, allowing animals to rapidly protect themselves from changes in their internal and external environment. However, the complex nature of these systems in mammals makes it difficult to determine how neuronal signaling influences the immune response. Here we show that serotonin, synthesized in Caenorhabditis elegans chemosensory neurons, modulates the immune response. Serotonin released from these cells acts, directly or indirectly, to regulate G-protein signaling in epithelial cells. Signaling in these cells is required for the immune response to infection by the natural pathogen Microbacterium nematophilum. Here we show that serotonin signaling suppresses the innate immune response and limits the rate of pathogen clearance. We show that C. elegans uses classical neurotransmitters to alter the immune response. Serotonin released from sensory neurons may function to modify the immune system in response to changes in the animal's external environment such as the availability, or quality, of food. PMID:24348250

  17. A 90-day safety study of genetically modified rice expressing Cry1Ab protein (Bacillus thuringiensis toxin) in Wistar rats

    DEFF Research Database (Denmark)

    Schrøder, Malene; Poulsen, Morten; Wilcks, Andrea

    2007-01-01

    An animal model for safety assessment of genetically modified foods was tested as part of the SAFOTEST project. In a 90-day feeding study on Wistar rats, the transgenic KMD1 rice expressing Cry1Ab protein was compared to its non-transgenic parental wild type, Xiushui 11. The KMD1 rice contained 15......, macroscopic and histopathological examinations were performed with only minor changes to report. The aim of the study was to use a known animal model in performance of safety assessment of a GM crop, in this case KMD1 rice. The results show no adverse or toxic effects of KMD1 rice when tested in the design...... used in this 90-day study. Nevertheless the experiences from this study lead to the overall conclusion that safety assessment for unintended effects of a GM crop cannot be done without additional test group(s)....

  18. Assembly of Modified Ferritin Proteins on Carbon Nanotubes and its Electrocatalytic Activity for Oxygen Reduction

    Science.gov (United States)

    Kim, Jae-Woo; Lillehei, Peter T.; Park, Cheol

    2012-01-01

    Highly effective dispersions of carbon nanotubes (CNTs) can be made using a commercially available buffer solution. Buffer solutions of 3-(N-morpholino)-propanesulfonic acid (MOPS), which consists of a cyclic ring with nitrogen and oxygen heteroatoms, a charged group, and an alkyl chain greatly enhance the dispersibility and stability of CNTs in aqueous solutions. Additionally, the ability of biomolecules, especially cationized Pt-cored ferritins, to adhere onto the well-dispersed CNTs in the aqueous buffer solution is also improved. This was accomplished without the use of surfactant molecules, which are detrimental to the electrical, mechanical, and other physical properties of the resulting products. The assembled Pt-cored ferritin proteins on the CNTs were used as an electrocatalyst for oxygen reduction

  19. Mesoporous Silicon with Modified Surface for Plant Viruses and Their Protein Particle Sensing

    Directory of Open Access Journals (Sweden)

    Kae Dal Kwack

    2008-10-01

    Full Text Available Changes in electric parameters of a mesoporous silicon treated by a plasma chemical etching with fluorine and hydrogen ions, under the adsorption of NEPO (Nematodetransmitted Polyhedral plant viruses such as TORSV (Tomato Ringspot Virus, GFLV (Grapevine Fan Leaf Virus and protein macromolecule from TORSV particles are described. The current response to the applied voltage is measured for each virus particle to investigate the material parameters which are sensitive to the adsorbed particles. The peculiar behaviors of the response are modeled by the current-voltage relationship in a MOSFET. This model explains the behavior well and the double gate model of the MOSFET informs that the mesoporous silicon is a highly sensitive means of detecting the viruses in the size range less than 50 nm.

  20. Awake intranasal insulin delivery modifies protein complexes and alters memory, anxiety, and olfactory behaviors.

    Science.gov (United States)

    Marks, David R; Tucker, Kristal; Cavallin, Melissa A; Mast, Thomas G; Fadool, Debra A

    2009-05-20

    The role of insulin pathways in olfaction is of significant interest with the widespread pathology of diabetes mellitus and its associated metabolic and neuronal comorbidities. The insulin receptor (IR) kinase is expressed at high levels in the olfactory bulb, in which it suppresses a dominant Shaker ion channel (Kv1.3) via tyrosine phosphorylation of critical N- and C-terminal residues. We optimized a 7 d intranasal insulin delivery (IND) in awake mice to ascertain the biochemical and behavioral effects of insulin to this brain region, given that nasal sprays for insulin have been marketed notwithstanding our knowledge of the role of Kv1.3 in olfaction, metabolism, and axon targeting. IND evoked robust phosphorylation of Kv1.3, as well as increased channel protein-protein interactions with IR and postsynaptic density 95. IND-treated mice had an increased short- and long-term object memory recognition, increased anxiolytic behavior, and an increased odor discrimination using an odor habituation protocol but only moderate change in odor threshold using a two-choice paradigm. Unlike Kv1.3 gene-targeted deletion that alters metabolism, adiposity, and axonal targeting to defined olfactory glomeruli, suppression of Kv1.3 via IND had no effect on body weight nor the size and number of M72 glomeruli or the route of its sensory axon projections. There was no evidence of altered expression of sensory neurons in the epithelium. In mice made prediabetic via diet-induced obesity, IND was no longer effective in increasing long-term object memory recognition nor increasing anxiolytic behavior, suggesting state dependency or a degree of insulin resistance related to these behaviors.

  1. Aspects of 6-[18F]fluoro-L-DOPA preparation: precursor synthesis, preparative HPLC purification and determination of radiochemical purity

    International Nuclear Information System (INIS)

    Fuechtner, F.; Angelberger, P.; Kvaternik, H.; Hammerschmidt, F.; Simovc, B. Peric; Steinbach, J.

    2002-01-01

    A modified method for the synthesis of the intermediate product N-Boc-3,4-di(Boc-O)-6-iodo-L-phenylalanine ethyl ester of the [ 18 F]FDOPA precursor preparation was developed. With the application of bis-(trifluoroacetoxy)-iodobenzene for the iodination step with elemental iodine the yield of the intermediate can be increased from 12% to 50-60%. By replacing silica-gel-based RP HPLC column by a polymer-based column for semi-preparative purification of [ 18 F]FDOPA from the reaction mixture the radiochemical purity of the final product can be increased up to >99%. For the determination of the radiochemical impurity [ 18 F]fluoride a HPLC method using a column with polymer-based RP material was introduced

  2. [The effect of hydrophobic surface properties of protein on its resistance to denaturation by organic solvents (using modified alpha-chymotrypsin as an example].

    Science.gov (United States)

    Kudriashova, E V; Belova, A B; Vinogradov, A A; Mozhaev, V V

    1994-03-01

    Catalytic activity of covalently modified alpha-chymotrypsin in water/cosolvent solutions was investigated. The stability of chymotrypsin increases upon modification with hydrophilic reagents, such as glyceraldehyde, pyrometallic and succinic anhydrides, and glucosamine. Correlation was observed between the protein's stability in organic solvents and the degree of hydrophilization of the protein's surface. The protein is the more stable, the higher are the modification degree and the hydrophilicity of the modifying residue. At a certain critical hydrophilization degree of chymotrypsin a limit of stability is achieved. The stabilization effect can be accounted for by the fact that the interaction between water molecules on the surface and protein's functional groups become stronger in the hydrophilized protein.

  3. Paleomagnetism and radiochemical age estimates for Late Brunhes polarity episodes

    International Nuclear Information System (INIS)

    Denham, C.R.; Anderson, R.F.; Bacon, M.P.

    1977-01-01

    Several reversed polarity magnetozones occur within deep-sea sediment core CH57-8 from the Greater Antilles Outer Ridge, within sediment of latest Pleistocene/Late Brunhes age. The uppermost reversed interval spanning 31 data points coincides with the X faunal zone of the Last Interglacial Period. Radiochemical dating of cores CH57-8 and KN25-4 has shown that all the reversed polarity magnetozones are significantly younger than the Brunhes/Matuyama boundary at 0.7 m.y.B.P. A variation of the excess 230 Th method was used, in which 210 Po and 238 U were the actual radionuclides measured. In a third core from the Mid-Atlantic Ridge, the 210 Po results were similar to those which others obtained earlier by direct 230 Th measurements. (Auth.)

  4. Development of robotic plasma radiochemical assays for positron emission tomography

    International Nuclear Information System (INIS)

    Alexoff, D.L.; Shea, C.; Fowler, J.S.; Gatley, S.J.; Schlyer, D.J.

    1995-01-01

    A commercial laboratory robot system (Zymate PyTechnology II Laboratory Automation System; Zymark Corporation, Hopkinton, MA) was interfaced to standard and custom laboratory equipment and programmed to perform rapid radiochemical analyses for quantitative PET studies. A Zymark XP robot arm was used to carry out the determination of unchanged (parent) radiotracer in plasma using only solid phase extraction methods. Robotic throughput for the assay of parent radiotracer in plasma is 4--6 samples/hour depending on the radiotracer. Robotic assays of parent compound in plasma were validated for the radiotracers [ 11 C]Benztropine, [ 11 C]cocaine, [ 11 C]clorgyline, [ 11 C]deprenyl, [ 11 C]methadone, [ 11 C]methylphenidate, [ 11 C]raclorpride, and [ 11 C]SR46349B. A simple robot-assisted methods development strategy has been implemented to facilitate the automation of plasma assays of new radiotracers

  5. Quantitative radio-chemical separation of calcium, strontium and barium

    International Nuclear Information System (INIS)

    Dupuis, M.C.; Dupuis, M.; Le Nagard, M.; Michot, H.

    1965-01-01

    A method for separation of Ca 45 , Sr 89 and Ba 140 has been developed for the radiochemical determination of these isotopes in a solution of fission with a large concentration of mineral salts. After removal of most fission products by solvent extraction (TTA-MIBK) at different pH, the alkaline earths are extracted from the aqueous phase at pH 9. After recovery with diluted hydrochloric acid, the three elements are adsorbed on cationic resin Dowex 50 and eluted sequentially with ammonium α - Hydroxy iso-butyrate using gradient concentration and pH. Ca 45 and Sr 89 are measured by β - counting and Ba 140 by γ spectrometry. The chemical yield approximates 80 per cent for calcium, and 70 per cent for strontium and barium. The decontamination factor is 10 5 for most fission products. Four separations can be performed in twenty hours. (authors) [fr

  6. Handling of Ammonium Nitrate Mother-Liquid Radiochemical Production - 13089

    International Nuclear Information System (INIS)

    Zherebtsov, Alexander; Dvoeglazov, Konstantine; Volk, Vladimir; Zagumenov, Vladimir; Zverev, Dmitriy; Tinin, Vasiliy; Kozyrev, Anatoly; Shamin, Dladimir; Tvilenev, Konstantin

    2013-01-01

    The aim of the work is to develop a basic technology of decomposition of ammonium nitrate stock solutions produced in radiochemical enterprises engaged in the reprocessing of irradiated nuclear fuel and fabrication of fresh fuel. It was necessary to work out how to conduct a one-step thermal decomposition of ammonium nitrate, select and test the catalysts for this process and to prepare proposals for recycling condensation. Necessary accessories were added to a laboratory equipment installation decomposition of ammonium nitrate. It is tested several types of reducing agents and two types of catalyst to neutralize the nitrogen oxides. It is conducted testing of modes of the process to produce condensation, suitable for use in the conversion of a new technological scheme of production. It is studied the structure of the catalysts before and after their use in a laboratory setting. It is tested the selected catalyst in the optimal range for 48 hours of continuous operation. (authors)

  7. Radiochemical analysis in the nuclear research establishment (KFA) Juelich, FRG

    International Nuclear Information System (INIS)

    Anon.

    1975-01-01

    KFA Juelich is one of the two great nuclear research centres of the Federal Republic of Germany. About 3700 employees including about 700 scientists are engaged in a great number of programs and projects belonging to six main fields of research and development: high temperature reactor and energy techniques; nuclear fusion; properties of materials; materials research; life and environment; methods. In the article the radiochemical analysis work of the former Central Institute of Analytical Chemistry and its two successors is described: activation analysis, application of tracer techniques, fission product analysis. Further on the irradiation facilities are described, a short survey is given on the instrumentation, and the future work is outlined. (T.G.)

  8. Computer aided design of piping for a radiochemical plant

    Energy Technology Data Exchange (ETDEWEB)

    Selvaraj, P G; Chandrasekhar, A; Chandrasekar, A V [Reprocessing Group, Indira Gandhi Centre for Atomic Research, Kalpakkam (India); Raju, R P; Mahudeeswaran, K V; Kumar, S V [Reprocessing Group, Bhabha Atomic Research Centre, Mumbai (India)

    1994-06-01

    In a radiochemical plant such as reprocessing plants, process equipment, storage tanks, liquid transfer systems and the associated pipe lines etc. are housed in series of concrete cells. Availability of limited cell space/volume, provision of various modes of liquid transfers with associated redundancies and instrumentation lines with standby alternatives increase the overall piping density. Designing such high density piping layout without interference is quite complex and needs lot of human efforts. This paper briefly describes development of computer codes for the entire scheme of design, drafting and fabrication of piping for nuclear fuel reprocessing plant. The general organisation of various programs, their functions, the complete sequence of the scheme and the flow of data are presented. High degree of reliability of each routine, considerable error checking facilities, marking legends on the drawings, provision for scaling in drafting and accuracy to the extent of one mm in layout design are some of the important features of this scheme. (author). 1 fig.

  9. 15th radiochemical conference: Booklet of abstracts and conference programme

    International Nuclear Information System (INIS)

    John, J.; Benes, P.; Kucera, J.; Havela, L.; Bartonicek, B.; Vobecky, M.; Krizova, V.; Kopicka, K.; Prasil, Z.

    2006-04-01

    The conference was structured as follows: Opening plenary lectures (6 lectures); Topic 1 - radionuclides in the environment, radioecology (22 verbal presentations (VPs), 23 poster presentations (PPs)); Topic 2 - nuclear analytical methods (22 VPs, 32 PPs); Topic 3 - chemistry of actinide and transactinide elements (8 VPs, 10 PPs); Topic 4 - radiation chemistry (9 VPs, 5 PPs); Topic 5 - production and application of radionuclides (17 VPs, 6 PPs); Topic 6 - separation methods, speciation (21 VPs, 23 PPs); Topic 7 - chemistry of nuclear fuel cycle, radiochemical problems in nuclear waste management (20 VPs, 16 PPs); Topic 8 - nuclear methods in medicine, radiopharmaceuticals, and radiodiagnostics, labelled compounds (8 VPs, 7 PPs); and Panels (2 introductions). (P.A.)

  10. The design of a new radiochemical laboratory complex

    International Nuclear Information System (INIS)

    Lewis, A.G.

    1984-01-01

    A brief account is given of the history and scope of Amersham International plc in the manufacture of radiopharmaceuticals and other labelled organic compounds, and radioactive sources. Extra facilities were needed and a new site was found, and contracts placed for new radiochemical laboratories. The two new laboratories, which are described in some detail, are intended as follows: (a) a Medical Products building for the production of a range of diagnostic kits for use in the treatment of thyroid and other disorders, the main isotope used being iodine-125; and (b) the Chemical Products building, for the development and manufacture of a wide range of organic compounds, which are labelled with either tritium or carbon-14. Particular emphasis is given to the description of the air conditioning and ventilation systems, the open work benches, and the special ventilated enclosures, and the drainage system. Planning for maximum flexibility is also stressed. (U.K.)

  11. Fast analysis procedure of radiochemical coordinat uptake for methotrexate

    International Nuclear Information System (INIS)

    Caston, J.D.; Kamen, B.A.

    1976-01-01

    Under this invention, a radio-chemical analysis is submitted to determine the concentration of methotrexate or its equivalents in analysis in a biological medium. The amounts taken up of the labelled compound and the known concentrations of the unlabelled compound to be determined are radio-isotopically related to a first system containing a pre-determined amount of the labelled compound and a pre-determined amount of the unlabelled compound. In a second system, identical to the first, save that the sample of the biological medium to be analyzed takes the place of the unlabelled compound, the amount of labelled compound taken up is determined radio-isotopically. The concentration of the compound in the sample is then determined by correlation of the labelled compound uptake determined in the second system with the relation determined in the first system. The radio-isotopic relations and determinations may be made by direct and sequential analytical techniques [fr

  12. Radiochemical neutron activation analysis of gold in geochemical samples

    International Nuclear Information System (INIS)

    Zilliacus, R.

    1983-01-01

    A fast method for the radiochemical neutron activation analysis of gold in geochemical samples is described. The method is intended for samples having background concentrations of gold. The method is based on the dissolution of samples with hydrofluoric acid and aqua regia followed by the dissolution of the fluorides with boric acid and hydrochloric acid. Gold is then adsorbed on activated carbon by filtrating the solution through a thin carbon layer. The activity measurements are carried out using a Ge(Li)-detector and a multichannel analyzer. The chemical yields of the separation determined by reirradiation vary between 60 and 90%. The detection limit of the method is 0.2 ng/g gold in rock samples. USGS standard rocks and exploration reference materials are analyzed and the results are presented and compared with literature data. (author)

  13. Radiochemical methods for studying lipase-catalyzed interesterification of lipids

    International Nuclear Information System (INIS)

    Schuch, R.; Mukherjee, K.D.

    1987-01-01

    Reactions involving lipase-catalyzed interesterification of lipids, which are of commendable interest in biotechnology, have been monitored and assayed by radiochemical methods using 14 C-labeled substrates. Medium chain (C 12 plus C 14 ) triacylglycerols were reacted in the presence of an immobilized lipase from Mucor miehei and hexane at 45 0 C with methyl [1- 14 C]oleate, [1- 14 C]oleic acid, [carboxyl- 14 C]trioleoylglycerol, [1- 14 C]octadecenyl alcohol, and [U- 14 C]glycerol, each of known specific activity. The reactions were monitored and the rate of interesterification determined by radio thin layer chromatography from the incorporation of radioactivity into acyl moieties of triacylglycerols (from methyl oleate, oleic acid, and trioleoylglycerol), alkyl moieties of wax esters (from octadecenyl alcohol), and into glycerol backbone of monoacylglycerols and diacylglycerols (from glycerol). (orig.)

  14. Research reactor FR2 - 20 years chemical and radiochemical measurements

    International Nuclear Information System (INIS)

    Feuerstein, H.; Graebner, H.; Oschinski, J.; Hoffmann, W.; Beyer, J.

    1986-09-01

    The FR2 has been a D 2 O cooled and moderated research reactor with a thermal output of 44 MW. It was in operation from 1961 to 1981. Because of the operating conditions of the reactor, only a small number of routine measurements were performed. For these however special techniques had to be developed. During the 20 years of operation a number of special events occured or have been observed, sometimes with very amazing results, e.g. the 'aceton effect'. This report describes the chemical and radiochemical conditions of the reactor systems, as well as the results of the surveilance work. Not described are measurements for the many experiments. The last chapter gives in a short form a description of the most unusual events and observations. (orig.) [de

  15. Radiochemical plasma salicylamide assay using ring-labeled tritiated salicylamide

    Energy Technology Data Exchange (ETDEWEB)

    Stella, V J; Varia, S A; Riedy, M

    1979-05-01

    A rat plasma salicylamide assay was developed using ring-labeled tritiated salicylamide, synthesized by reacting salicylamide with tritium oxide in the presence of heptafluorobutyric acid. The reaction yielded /sup 3/H-salicylamide of specific activity up to 8.41 mCi/mmole, 60% yield. Plasma containing /sup 3/H-salicylamide and its metabolites was extracted with a toluene-based scintillation fluid, which was subsequently counted. Specificity for free salicylamide was demonstrated by radiochemical and standard fluorescence plasma salicylamide level-time curves. Specificity resulted from nonextraction of the salicylamide sulfate and glucuronide metabolites. Sulfatase and beta-glucuronidase treatment allowed the analysis of plasma sulfate and glucuronide conjugates as free salicylamide. This procedure should be effective for the analysis of salicylamide and its metabolites in the presence of similar phenolic compounds.

  16. Radiochemical investigations on the solubility of molybdatophosphate in phosphate determination

    International Nuclear Information System (INIS)

    Noack, S.

    1975-01-01

    The solubility of various molybdatophosphates was determined under the conditions of a gravimetric phosphate determination by radiochemical means by labelling PO 4 3- with P-32. Starting with various conditions for phosphate determination via the molybdatophosphate of quinoline, 8-hydroxyquinoline, dimorpholino ethane, N,N,N',N'-tetrakis-β-hydroxypropyl ethylene diamine and N,N,N',N'-tetrakis-β-hydroxybutyl ethylene diamine, a general working rule was developed to determine the solubility. Taking the example of quinoline molybdatophosphates, a series of influencing factors - work, concentration and measuring parameters - were investigated in order to be able to limit the reliability region of the gravimetric phosphate determination. Depending on the conditions, the measured solubilities were between 10 -10 and 10 -6 Mol/l, the corresponding degrees of precipitation between 99.0 and 99.9999%. Apparent solubility products were calculated for the different molybdatophosphates using computer programmes especially developed for this purpose. (orig./RB) [de

  17. A rapid, simple method for obtaining radiochemically pure hepatic heme

    International Nuclear Information System (INIS)

    Bonkowski, H.L.; Bement, W.J.; Erny, R.

    1978-01-01

    Radioactively-labelled heme has usually been isolated from liver to which unlabelled carrier has been added by long, laborious techniques involving organic solvent extraction followed by crystallization. A simpler, rapid method is devised for obtaining radiochemically-pure heme synthesized in vivo in rat liver from delta-amino[4- 14 C]levulinate. This method, in which the heme is extracted into ethyl acetate/glacial acetic acid and in which porphyrins are removed from the heme-containing organic phase with HCl washes, does not require addition of carrier heme. The new method gives better heme recoveries than and heme specific activities identical to, those obtained using the crystallization method. In this new method heme must be synthesized from delta-amino[4- 14 C]levulinate; it is not satisfactory to use [2- 14 C]glycine substrate because non-heme counts are isolated in the heme fraction. (Auth.)

  18. Radiochemical analyses of several spent fuel Approved Testing Materials

    International Nuclear Information System (INIS)

    Guenther, R.J.; Blahnik, D.E.; Wildung, N.J.

    1994-09-01

    Radiochemical characterization data are described for UO 2 and UO 2 plus 3 wt% Gd 2 O 3 commercial spent nuclear fuel taken from a series of Approved Testing Materials (ATMs). These full-length nuclear fuel rods include MLA091 of ATM-103, MKP070 of ATM-104, NBD095 and NBD131 of ATM-106, and ADN0206 of ATM-108. ATMs 103, 104, and 106 were all irradiated in the Calvert Cliffs Nuclear Power Plant (Reactor No.1), a pressurized-water reactor that used fuel fabricated by Combustion Engineering. ATM-108 was part of the same fuel bundle designed as ATM-105 and came from boiling-water reactor fuel fabricated by General Electric and irradiated in the Cooper Nuclear Power Plant. Rod average burnups and expected fission gas releases ranged from 2,400 to 3,700 GJ/kgM. (25 to 40 Mwd/kgM) and from less than 1% to greater than 10%, respectively, depending on the specific ATM. The radiochemical analyses included uranium and plutonium isotopes in the fuel, selected fission products in the fuel, fuel burnup, cesium and iodine on the inner surfaces of the cladding, 14 C in the fuel and cladding, and analyses of the gases released to the rod plenum. Supporting examinations such as fuel rod design and material descriptions, power histories, and gamma scans used for sectioning diagrams are also included. These ATMs were examined as part of the Materials Characterization Center Program conducted at Pacific Northwest Laboratory provide a source of well-characterized spent fuel for testing in support of the US Department of Energy Office of Civilian Radioactive Waste Management Program

  19. Whey protein isolate modified by transglutaminase aggregation and emulsion gel properties

    Science.gov (United States)

    Qi, Weiwei; Chen, Chong; Liu, Mujun; Yu, Guoping; Cai, Xinghang; Guo, Peipei; Yao, Yuxiu; Mei, Sijie

    2015-07-01

    Whey protein isolate and commercial soybean salad oil were used to produce the WPI emulsion dispersions. The properties of TG-catalyzed emulsion gelation produced from WPI emulsion dispersions were investigated by the amount of TG, temperature, pH and reaction time. Specifically, the texture properties (hardness and springiness), water-holding capacity and rheological properties (G' and G") were assessed. The result of Orthogonal tests showed WPI emulsion can form better hardness and springiness gel when the ratio of TG and WPI was 20U/g, pH 7.5, treatment temperature and time were 50°C and 3 h, respectively. The microstructure of TG emulsion gels was more compact, gel pore is smaller, distribution more uniform, the oil droplets size smaller compared with untreated emulsion gels. Compared to the control of rheological properties, G' and G" were significantly increased and G' > G", results showed that the gel was solid state, and TG speeded up the process of gelation.

  20. Dual Mode Fluorophore-Doped Nickel Nitrilotriacetic Acid-Modified Silica Nanoparticles Combine Histidine-Tagged Protein Purification with Site-Specific Fluorophore Labeling

    OpenAIRE

    Kim, Sung Hoon; Jeyakumar, M.; Katzenellenbogen, John A.

    2007-01-01

    We present the first example of a fluorophore-doped nickel chelate surface- modified silica nanoparticle that functions in a dual mode, combining histidine-tagged protein purification with site-specific fluorophore labeling. Tetramethylrhodamine (TMR)-doped silica nanoparticles, estimated to contain 700–900 TMRs per ca. 23-nm particle, were surface modified with nitrilotriacetic acid (NTA), producing TMR-SiO2-NTA-Ni+2. Silica-embedded TMR retains very high quantum yield, is resistant to quenc...

  1. Immunotoxicological Evaluation of Genetically Modified Rice Expressing Cry1Ab/Ac Protein (TT51-1) by a 6-Month Feeding Study on Cynomolgus Monkeys

    OpenAIRE

    Tan, Xiaoyan; Zhou, Xiaobing; Tang, Yao; Lv, Jianjun; Zhang, Lin; Sun, Li; Yang, Yanwei; Miao, Yufa; Jiang, Hua; Chen, Gaofeng; Huang, Zhiying; Wang, Xue

    2016-01-01

    The present study was performed to evaluate the food safety of TT51-1, a new type of genetically modified rice that expresses the Cry1Ab/Ac protein (Bt toxin) and is highly resistant to most lepidopteran pests. Sixteen male and 16 female cynomolgus monkeys were randomly divided into four groups: conventional rice (non-genetically modified rice, non-GM rice), positive control, 17.5% genetically modified rice (GM rice) and 70% GM rice. Monkeys in the non-GM rice, positive control, and GM rice g...

  2. Automation of column-based radiochemical separations. A comparison of fluidic, robotic, and hybrid architectures

    Energy Technology Data Exchange (ETDEWEB)

    Grate, J.W.; O' Hara, M.J.; Farawila, A.F.; Ozanich, R.M.; Owsley, S.L. [Pacific Northwest National Laboratory, Richland, WA (United States)

    2011-07-01

    Two automated systems have been developed to perform column-based radiochemical separation procedures. These new systems are compared with past fluidic column separation architectures, with emphasis on using disposable components so that no sample contacts any surface that any other sample has contacted, and setting up samples and columns in parallel for subsequent automated processing. In the first new approach, a general purpose liquid handling robot has been modified and programmed to perform anion exchange separations using 2 mL bed columns in 6 mL plastic disposable column bodies. In the second new approach, a fluidic system has been developed to deliver clean reagents through disposable manual valves to six disposable columns, with a mechanized fraction collector that positions one of four rows of six vials below the columns. The samples are delivered to each column via a manual 3-port disposable valve from disposable syringes. This second approach, a hybrid of fluidic and mechanized components, is a simpler more efficient approach for performing anion exchange procedures for the recovery and purification of plutonium from samples. The automation architectures described can also be adapted to column-based extraction chromatography separations. (orig.)

  3. An autoantibody against Nε-(carboxyethyl)lysine (CEL): Possible involvement in the removal of CEL-modified proteins by macrophages

    International Nuclear Information System (INIS)

    Mera, Katsumi; Nagai, Ryoji; Takeo, Kazuhiro; Izumi, Miyoko; Maruyama, Toru; Otagiri, Masaki

    2011-01-01

    Highlights: → A higher amount of autoantibody against CEL than that of other AGEs was observed in human plasma. → The purified human anti-CEL autoantibody specifically reacted with CEL. → Anti-CEL antibody accelerated the uptake of 125 I-CEL-HSA by macrophage in vitro. → Endocytic uptake of 125 I-CEL-HSA by mice liver was accelerated in the presence of anti-CEL antibody. -- Abstract: Advanced glycation end products (AGEs) are believed to play a significant role in the development of diabetic complications. In this study, we measured the levels of autoantibodies against several AGE structures in healthy human plasma and investigated the physiological role of the autoantibodies. A high titer of the autoantibody against N ε -(carboxyethyl)lysine (CEL) was detected in human plasma compared with other AGE structures such as CML and pentosidine. The purified human anti-CEL autoantibody reacted with CEL-modified human serum albumin (CEL-HSA), but not CML-HSA. A rabbit polyclonal anti-CEL antibody, used as a model autoantibody against CEL, accelerated the uptake of CEL-HSA by macrophages, but did not enhance the uptake of native HSA. Furthermore, when 125 I-labeled CEL-HSA was injected into the tail vein of mice, accumulation of 125 I-CEL-HSA in the liver was accelerated by co-injection of the rabbit anti-CEL antibody. These results demonstrate that the autoantibody against CEL in plasma may play a role in the macrophage uptake of CEL-modified proteins.

  4. O-GlcNAc-specific antibody CTD110.6 cross-reacts with N-GlcNAc2-modified proteins induced under glucose deprivation.

    Directory of Open Access Journals (Sweden)

    Takahiro Isono

    Full Text Available Modification of serine and threonine residues in proteins by O-linked β-N-acetylglucosamine (O-GlcNAc glycosylation is a feature of many cellular responses to the nutritional state and to stress. O-GlcNAc modification is reversibly regulated by O-linked β-N-acetylglucosamine transferase (OGT and β-D-N-acetylglucosaminase (O-GlcNAcase. O-GlcNAc modification of proteins is dependent on the concentration of uridine 5'-diphospho-N-acetylglucosamine (UDP-GlcNAc, which is a substrate of OGT and is synthesized via the hexosamine biosynthetic pathway. Immunoblot analysis using the O-GlcNAc-specific antibody CTD110.6 has indicated that glucose deprivation increases protein O-GlcNAcylation in some cancer cells. The mechanism of this paradoxical phenomenon has remained unclear. Here we show that the increased glycosylation induced by glucose deprivation and detected by CTD110.6 antibodies is actually modification by N-GlcNAc(2, rather than by O-GlcNAc. We found that this induced glycosylation was not regulated by OGT and O-GlcNAcase, unlike typical O-GlcNAcylation, and it was inhibited by treatment with tunicamycin, an N-glycosylation inhibitor. Proteomics analysis showed that proteins modified by this induced glycosylation were N-GlcNAc(2-modified glycoproteins. Furthermore, CTD110.6 antibodies reacted with N-GlcNAc(2-modified glycoproteins produced by a yeast strain with a ts-mutant of ALG1 that could not add a mannose residue to dolichol-PP-GlcNAc(2. Our results demonstrated that N-GlcNAc(2-modified glycoproteins were induced under glucose deprivation and that they cross-reacted with the O-GlcNAc-specific antibody CTD110.6. We therefore propose that the glycosylation status of proteins previously classified as O-GlcNAc-modified proteins according to their reactivity with CTD110.6 antibodies must be re-examined. We also suggest that the repression of mature N-linked glycoproteins due to increased levels of N-GlcNAc(2-modified proteins is a newly

  5. Colonic vascular conductance increased by Daikenchuto via calcitonin gene-related peptide and receptor-activity modifying protein 1.

    Science.gov (United States)

    Kono, Toru; Koseki, Takashi; Chiba, Shinichi; Ebisawa, Yoshiaki; Chisato, Naoyuki; Iwamoto, Jun; Kasai, Shinichi

    2008-11-01

    Daikencyuto (DKT) is a traditional Japanese medicine (Kampo) and is a mixture of extract powders from dried Japanese pepper, processed ginger, ginseng radix, and maltose powder and has been used as the treatment of paralytic ileus. DKT may increase gastrointestinal motility by an up-regulation of the calcitonin gene-related peptide (CGRP). CGRP is also the most powerful vasoactive substance. In the present study, we investigated whether DKT has any effect on the colonic blood flow in rats. Experiments were performed on fasted anesthetized and artificially ventilated Wistar rats. Systemic mean arterial blood pressure and heart rate were recorded. Red blood cell flux in colonic blood flow was measured using noncontact laser tissue blood flowmetry, and colonic vascular conductance (CVC) was calculated as the ratio of flux to mean arterial blood pressure. We examined four key physiological mechanisms underlying the response using blocker drugs: CGRP1 receptor blocker (CGRP(8-37)), nitric oxide synthase inhibitor, vasoactive intestinal polypeptide (VIP) receptor blocker ([4-Cl-DPhe6, Leu17]-VIP), and substance P receptor blocker (spantide). Reverse transcription-polymerase chain reaction was used for the detection of mRNA of calcitonin receptor-like receptor, receptor-activity modifying protein 1, the component of CGRP 1 receptor and CGRP. After laparotomy, a cannula was inserted into the proximal colon to administer the DKT and to measure CVC at the distal colon. Intracolonal administration of DKT (10, 100, and 300 mg/kg) increased CVC (basal CVC, 0.10 mL/mmHg) from the first 15-min observation period (0.14, 0.17, and 0.17 mL/mmHg, respectively) and with peak response at either 45 min (0.17 mL/mmHg by 10 mg/kg), or 75 and 60 min (0.23 and 0.21 mL/mmHg by 100 and 300 mg/kg, respectively). CGRP(8-37) completely abolished the DKT-induced hyperemia, whereas nitric oxide synthase inhibitor partially attenuated the DKT-induced hyperemia. [4-Cl-DPhe6, Leu17]-VIP and spantide

  6. Determination of radiochemical purity and pharmacokinetic parameters of sup(99m)Tc-sulphur colloid and sup(99m)Tc-tin colloid

    International Nuclear Information System (INIS)

    Jovanovic, V.; Konstantinovska, D.; Milivojevic, K.; Bzenic, J.

    1981-01-01

    Labelling yield and radiochemical purity, higher than 95%, of sup(99m)Tc-colloid preparations were determined by using the paper chromatography method. Less than 3% of labelled citric acid, added to the preparation as a buffer solution, has been found in sup(99m)Tc-sulphur colloid. High radiochemical purity and optimum size of colloid particles has also been proved by biodistribution studies on experimental animals. The analysis performed has shown that more than 50% of sup(99m)Tc-colloid preparations excreted by urine is sup(99m)TcO - , the remaining past 50% being protein bound sup(99m)Tc. Biological half-time of excretion of the fast phase is the same for both preparations, i.e. 10 min, while for the slow component it is 120 min in sup(99m)Tc-S-colloid and 160 min in sup(99m)Tc-Sn colloid. (orig.) [de

  7. Radioactive preparations. Determination of radiochemical purity by thin-layer chromatography

    International Nuclear Information System (INIS)

    1986-01-01

    The standard sets the data which must be attached to every sample, and the equipment, chemicals and auxiliary substances used in the determination of radiochemical purity of substances by chromatography. Described are preparation of the sample, the procedure of sample deposition, the development, drying and detection of the radioactive preparation. The qualitative and quantitative assessment of the radiochromatogram is described as are the calculation of radiochemical purity and the determination of the reproducibility of measurement of radiochemical purity of radioactive preparations. (E.S.)

  8. Activation of extrasynaptic, but not synaptic, NMDA receptors modifies amyloid precursor protein expression pattern and increases amyloid-ß production.

    Science.gov (United States)

    Bordji, Karim; Becerril-Ortega, Javier; Nicole, Olivier; Buisson, Alain

    2010-11-24

    Calcium is a key mediator controlling essential neuronal functions depending on electrical activity. Altered neuronal calcium homeostasis affects metabolism of amyloid precursor protein (APP), leading to increased production of β-amyloid (Aβ), and contributing to the initiation of Alzheimer's disease (AD). A linkage between excessive glutamate receptor activation and neuronal Aβ release was established, and recent reports suggest that synaptic and extrasynaptic NMDA receptor (NMDAR) activation may have distinct consequences in plasticity, gene regulation, and neuronal death. Here, we report for the first time that prolonged activation of extrasynaptic NMDAR, but not synaptic NMDAR, dramatically increased the neuronal production of Aβ. This effect was preceded by a shift from APP695 to Kunitz protease inhibitory domain (KPI) containing APPs (KPI-APPs), isoforms exhibiting an important amyloidogenic potential. Conversely, after synaptic NMDAR activation, we failed to detect any KPI-APP expression and neuronal Aβ production was not modified. Calcium imaging data showed that intracellular calcium concentration after extrasynaptic NMDAR stimulation was lower than after synaptic activation. This suggests distinct signaling pathways for each pool of receptors. We found that modification of neuronal APP expression pattern triggered by extrasynaptic NMDAR activation was regulated at an alternative splicing level involving calcium-/calmodulin-dependent protein kinase IV, but overall APP expression remained identical. Finally, memantine dose-dependently inhibited extrasynaptic NMDAR-induced KPI-APPs expression as well as neuronal Aβ release. Altogether, these data suggest that a chronic activation of extrasynaptic NMDAR promotes amyloidogenic KPI-APP expression leading to neuronal Aβ release, representing a causal risk factor for developing AD.

  9. Determination of hydrophobic coenzyme a esters and other lipids using a biosensor comprising a modified coenzyme a- and acyl-coa binding protein (acbp)

    DEFF Research Database (Denmark)

    2002-01-01

    , food and feed preparations, tissue extracts, acyl-CoA synthetase reaction media and various laboratory conditions using a modified Coenzyme A- and acyl-CoA binding protein (ACBP) is provided. Furthermore the invention relates to a construct comprising a peptide and a signal moiety for performing...

  10. The effect of modifying dietary protein and carbohydrate in weight loss on arterial compliance and postprandial lipidemia in overweight women with polycystic ovary syndrome.

    Science.gov (United States)

    Moran, Lisa J; Noakes, Manny; Clifton, Peter M; Norman, Robert J

    2010-11-01

    In overweight women with polycystic ovary syndrome, weight loss improves arterial compliance and postprandial lipidemia. Modifying dietary carbohydrate or protein in weight loss provided similar improvements in arterial compliance and postprandial lipidemia. Copyright © 2010 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  11. 40 CFR 174.505 - Bacillus thuringiensis modified Cry3A protein (mCry3A) in corn; exemption from the requirement of...

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Bacillus thuringiensis modified Cry3A protein (mCry3A) in corn; exemption from the requirement of a tolerance. 174.505 Section 174.505 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) PESTICIDE PROGRAMS PROCEDURES AND REQUIREMENTS FOR PLANT-INCORPORATED PROTECTANTS...

  12. The Generation of Turnip Crinkle Virus-Like Particles in Plants by the Transient Expression of Wild-Type and Modified Forms of Its Coat Protein.

    Science.gov (United States)

    Saunders, Keith; Lomonossoff, George P

    2015-01-01

    Turnip crinkle virus (TCV), a member of the genus carmovirus of the Tombusviridae family, has a genome consisting of a single positive-sense RNA molecule that is encapsidated in an icosahedral particle composed of 180 copies of a single type of coat protein. We have employed the CPMV-HT transient expression system to investigate the formation of TCV-like particles following the expression of the wild-type coat protein or modified forms of it that contain either deletions and/or additions. Transient expression of the coat protein in plants results in the formation of capsid structures that morphologically resemble TCV virions (T = 3 structure) but encapsidate heterogeneous cellular RNAs, rather than the specific TCV coat protein messenger RNA. Expression of an amino-terminal deleted form of the coat protein resulted in the formation of smaller T = 1 structures that are free of RNA. The possibility of utilizing TCV as a carrier for the presentation of foreign proteins on the particle surface was also explored by fusing the sequence of GFP to the C-terminus of the coat protein. The expression of coat protein-GFP hybrids permitted the formation of VLPs but the yield of particles is diminished compared to the yield obtained with unmodified coat protein. Our results confirm the importance of the N-terminus of the coat protein for the encapsidation of RNA and show that the coat protein's exterior P domain plays a key role in particle formation.

  13. Radiochemical search for neutron-rich isotopes of element 107

    International Nuclear Information System (INIS)

    Schaedel, M.

    1987-01-01

    Recent mass calculations have indicated that there is a region of deformed nuclei around neutron number N=162 that is especially stable against spontaneous fission. Barrier heights of about 5 MeV for Z = 107 nuclides can be extrapolated. To search for new, neutron-rich isotopes of element 107 in radiochemical experiments with 254 Es as a target an on-line chemical separation of element 107 (EKA-Rhenium), especially from the actinide elements is needed. An on-line gas-phase chemistry was developed with the homolog Re based on the volatility of the oxide which is transported in an O 2 containing atmosphere along a temperature gradient in a quartz tube and is condensed onto a thin Ta coated Ni-foil. The authors applied this technique in two series of experiments with their rotating wheel on-line gas-phase chemistry apparatus at the 88-inch cyclotron where they irradiated 254 Es as a target with 93 MeV and 96 MeV 16 O ions to search for 266 107. The assignment of the observed alpha events between 8 and 9 MeV to possibly (1) non actinide contaminants like 212 Po, (2) known isotopes of heavy elements like 261 105, or (3) a new isotope will be discussed

  14. Development of a radiochemical sensor. Part I: Feasibility study

    Energy Technology Data Exchange (ETDEWEB)

    Tarancon, A. [Departament de Quimica Analitica, Facultat de Quimica, Universitat de Barcelona, C/ Marti Franques 1, 08028 Barcelona (Spain); Garcia, J.F. [Departament de Pintura, Facultat de Belles Arts, Universitat de Barcelona, C/ Pau Gargallo 4, 08028 Barcelona (Spain)]. E-mail: jfgarcia@apolo.qui.ub.es; Rauret, G. [Departament de Quimica Analitica, Facultat de Quimica, Universitat de Barcelona, C/ Marti Franques 1, 08028 Barcelona (Spain)

    2005-05-04

    The evolution of nuclear activities and criteria for radiation protection have led to a continuous increase in measures to monitor and control the environment and therefore in the number of determinations required for such purposes. Classical analytical procedures are time-consuming, labor-intense and generate a large amount of waste. The alternative use of sensors for such determinations has seen very limited development. The present study focuses on the evaluation of the behavior of a prototype radiochemical sensor for liquid effluents. The sensor is based on a receptor made of a plastic scintillator and is capable of continuous, on-time and accurate remote quantification of the activity of alpha, beta and beta-gamma emitters. Low-level active solutions of {sup 90}Sr/{sup 90}Y, {sup 238}Pu, {sup 134}Cs and {sup 60}Co in matrices of groundwater, seawater and drinking water were quantified with prediction errors lower than 10% in most cases. The study also yields information about light generation and transmission and transductor configuration that will be useful in the design of future versions of this sensor.

  15. Planning for maintenance in radiochemical facilities [Paper No.: VB-2

    International Nuclear Information System (INIS)

    Balasubramanian, G.R.

    1981-01-01

    Reprocessing facilities in the earlier stages of development were planned mainly based on the concept of direct maintenance in view of the inherent advantage of man-machine interface and initial savings in the investment costs. With the mechanical processes finding a firm place in head-end operation and increase in down time necessary for elaborate decontamination efforts even for a minor modification has led to the review of the concept. For the same reason, the recent plants are based on the concept of harmonious blend of both direct and remote maintenance. The paper describes the planning needed from consideration of various aspects related to such concepts of maintenance during different phases of such type of facilities, highlighting some of the tools and special equipments to be developed for this purpose. A brief description of recent development in the field of remote maintenance is also given. Though the basic hot facility of reference is the one of reprocessing fast reactor fuels, the concepts and systems discussed are equally applicable to other radiochemical and radiometallurgical facilities also. (author)

  16. Waste treatment at the Radiochemical Engineering Development Center

    International Nuclear Information System (INIS)

    Brunson, R.R.; Bond, W.D.; Chattin, F.R.; Collins, R.T.; Sullivan, G.R.; Wiles, R.H.

    1997-01-01

    At the Radiochemical Engineering Development Center (REDC) irradiated targets are processed for the recovery of valuable radioisotopes, principally transuranium nuclides. A system was recently installed for treating the various liquid alkaline waste streams for removal of excess radioactive contaminants at the REDC. Radionuclides that are removed will be stored as solids and thus the future discharge of radionuclides to liquid low level waste tank storage will be greatly reduced. The treatment system is of modular design and is installed in a hot cell (Cubicle 7) in Building 7920 at the REDC where preliminary testing is in progress. The module incorporates the following: (1) a resorcinol-formaldehyde resin column for Cs removal, (2) a cross flow filtration unit for removal of rare earths and actinides as hydroxide, and (3) a waste solidification unit. Process flowsheets for operation of the module, key features of the module design, and its computer-assisted control system are presented. Good operability of the cross flow filter system is mandatory to the successful treatment of REDC wastes. Results of tests to date on the operation of the filter in its slurry collection mode and its slurry washing mode are presented. These tests include the effects of entrained organic solvent in the waste stream feed to the filter

  17. Remote sampling of process fluids in radiochemical plants

    International Nuclear Information System (INIS)

    Sengar, P.B.; Bhattacharya, R.; Ozarde, P. D.; Rana, D.S.

    1990-01-01

    Sampling of process fluids, continuous or periodic, is an essential requirement in any chemical process plant, so as to keep a control on process variables. In a radiochemical plant the task of taking and conveying the samples is a very tricky affair. This is due to the fact that neither the vessels/equipment containing radioactive effluents can be approached for manual sampling nor sampled fluids can be handled directly. The problems become more accute with higher levels of radioactivity. As such, inovative systems have to be devised to obtain and handle the raioactive samples employing remote operations. The remote sampling system developed in this Division has some of the unique features such as taking only requisite amount of samples in microlitre range, practically maintenance free design, avoidence of excess radioactive fluids coming out of process systems, etc. The paper describes in detail the design of remote sampling system and compares the same with existing systems. The design efforts are towards simplicity in operation, obtaining homogenised representative samples and highly economical on man-rem expenditure. The performance of a prototype system has also been evaluated. (author). 3 refs

  18. MAGGIE Component 1: Identification and Purification of Native and Recombinant Multiprotein Complexes and Modified Proteins from Pyrococcus furiosus

    Energy Technology Data Exchange (ETDEWEB)

    Adams, Michael W. [University of Georgia; W. W. Adams, Michael

    2014-01-07

    Virtualy all cellular processes are carried out by dynamic molecular assemblies or multiprotein complexes (PCs), the composition of which is largely unknown. Structural genomics efforts have demonstrated that less than 25% of the genes in a given prokaryotic genome will yield stable, soluble proteins when expressed using a one-ORF-at-a-time approach. We proposed that much of the remaining 75% of the genes encode proteins that are part of multiprotein complexes or are modified post-translationally, for example, with metals. The problem is that PCs and metalloproteins (MPs) cannot be accurately predicted on a genome-wide scale. The only solution to this dilemma is to experimentally determine PCs and MPs in biomass of a model organism and to develop analytical tools that can then be applied to the biomass of any other organism. In other words, organisms themselves must be analyzed to identify their PCs and MPs: “native proteomes” must be determined. This information can then be utilized to design multiple ORF expression systems to produce recombinant forms of PCs and MPs. Moreover, the information and utility of this approach can be enhanced by using a hyperthermophile, one that grows optimally at 100°C, as a model organism. By analyzing the native proteome at close to 100 °C below the optimum growth temperature, we will trap reversible and dynamic complexes, thereby enabling their identification, purification, and subsequent characterization. The model organism for the current study is Pyrococcus furiosus, a hyperthermophilic archaeon that grows optimally at 100°C. It is grown up to 600-liter scale and kg quantities of biomass are available. In this project we identified native PCs and MPs using P. furiosus biomass (with MS/MS analyses to identify proteins by component 4). In addition, we provided samples of abundant native PCs and MPs for structural characterization (using SAXS by component 5). We also designed and evaluated generic bioinformatics and

  19. Proceedings of the Tripartite Seminar on Nuclear Material Accounting and Control at Radiochemical Plants

    International Nuclear Information System (INIS)

    1999-01-01

    The problems of creation and operation of nuclear materials (NM) control and accounting systems and their components at radiochemical plants were discussed in seminar during November 2-6 of 1998. There were 63 Russian and 25 foreign participants in seminar. The seminar programme includes following sessions and articles: the aspects of State NM control and accountancy; NM control and accounting in radiochemical plants and at separate stages of reprocessing of spent nuclear fuel and irradiated fuel elements of commercial reactors; NM control and accountancy in storage facilities of radiochemical plants; NM control and accounting computerization, material balance assessment, preparation of reports; qualitative and quantitative measurements in NM control and accounting at radiochemical plants destructive analysis techniques [ru

  20. Automated radiochemical synthesis and biodistribution of [11C]l-α-acetylmethadol ([11C]LAAM)

    International Nuclear Information System (INIS)

    Sai, Kiran Kumar Solingapuram; Fan, Jinda; Tu, Zhude; Zerkel, Patrick; Mach, Robert H.; Kharasch, Evan D.

    2014-01-01

    Long-acting opioid agonists methadone and l-α-acetylmethadol (LAAM) prevent withdrawal in opioid-dependent persons. Attempts to synthesize [ 11 C]-methadone for PET evaluation of brain disposition were unsuccessful. Owing, however, to structural and pharmacologic similarities, we aimed to develop [ 11 C]LAAM as a PET ligand to probe the brain exposure of long-lasting opioids in humans. This manuscript describes [ 11 C]LAAM synthesis and its biodistribution in mice. The radiochemical synthetic strategy afforded high radiochemical yield, purity and specific activity, thereby making the synthesis adaptable to automated modules. - Highlights: • Radiochemical synthesis of opioid [ 11 C]l-α-acetylmethadol (LAAM) described for the first time. • High radiochemical yield, purity and specific activity. • Easily reproducible and adaptable synthesis to any C-11 automated modules. • [ 11 C]LAAM utility as a PET radiopharmaceutical for assessing brain penetration

  1. Investigations of radiochemical methods for the platinum group metals for NAA

    International Nuclear Information System (INIS)

    Tredoux, M.

    A radiochemical procedure for the determination of the platinum group metals and gold is outlined in this report. The sample is irradiated, treated with acids and passed through anion-exchange columns before being determined by gamma spectrometry

  2. Radiochemical and biological control of metaiodobenzyl-guanidine (MIBG) labeled with 131I

    International Nuclear Information System (INIS)

    Barboza, M.R.F.F. de; Muramoto, E.; Colturato, M.T.; Silva Valente Goncalves, R. da; Pereira, N.P.S. de; Almeida, M.A.T.M. de; Silva, C.P.G. da.

    1988-07-01

    This study shows the standardization of the radiochemical control of MIBG - 131 I in eletrophoretic system and also the biological control in Wistar rat for a period of time, not longer than 60 minutes after tracer administration. (author) [pt

  3. The use of radiochemical analysis for detecting biotracers of food radioactive contamination in Cherkasy Region

    International Nuclear Information System (INIS)

    Matvyijenko, D.G.

    2003-01-01

    Stable biotracers of radioactive contamination according to the findings of analytical control of the foodstuffs was determined. The use of radiochemical analysis for determining the activity of the foodstuffs and water (Sr-90, Cs-137) was evaluated

  4. First application of a microsphere-based immunoassay to the detection of genetically modified organisms (GMOs): quantification of Cry1Ab protein in genetically modified maize.

    Science.gov (United States)

    Fantozzi, Anna; Ermolli, Monica; Marini, Massimiliano; Scotti, Domenico; Balla, Branko; Querci, Maddalena; Langrell, Stephen R H; Van den Eede, Guy

    2007-02-21

    An innovative covalent microsphere immunoassay, based on the usage of fluorescent beads coupled to a specific antibody, was developed for the quantification of the endotoxin Cry1Ab present in MON810 and Bt11 genetically modified (GM) maize lines. In particular, a specific protocol was developed to assess the presence of Cry1Ab in a very broad range of GM maize concentrations, from 0.1 to 100% [weight of genetically modified organism (GMO)/weight]. Test linearity was achieved in the range of values from 0.1 to 3%, whereas fluorescence signal increased following a nonlinear model, reaching a plateau at 25%. The limits of detection and quantification were equal to 0.018 and 0.054%, respectively. The present study describes the first application of quantitative high-throughput immunoassays in GMO analysis.

  5. Natural radioprotection of cells. Radiochemical, biochemical and clinical aspects

    International Nuclear Information System (INIS)

    Revesz, L.

    1982-01-01

    Recent investigations on natural radioprotection are reviewed, carried out with human fibroblast strains and lymphoblastoid cell lines derived from individuals heterozygous or homozygous with regard to an inborn error in glutathione synthetase activity. The cells have a decreased content of non-protein bound sulphydryls and lack specifically glutathione either almost totally, or have a glutathione level about fifty per cent of the normal. Using single-strand DNA breaks as the end-point of the radiation effect, oxygen and misonidazole fail to enhance the radiosensitivity of cells deficient in glutathione, and OER and DMF close to unity are calculated. Substitution of glutathione deficiency by treatment with different radioprotective aminothiols decreases the sensitivity of the cells, and makes them susceptible to the oxygen effect. In glutathione deficient cells, rejoining of the single strand breaks proceeds at a normal rate after anoxic radiation exposure, but is inhibited after oxic exposure. Using clonogenic survival as the end-point for the radiation effect, sensitization of glutathione deficient cells is again greatly decreased, but OER and DMF differ significantly from unity. The data were interpreted to indicate that endogenous glutathione or some exogenous aminothiols repair radiation induced radicals in key target molecules by hydrogen transfer in a competition with oxygen and/or misonidazole which permanent the damage. The outcome of the radical reactions will be modified by further post-irradiation enzymatic repair processes of which at least one is glutathione dependent. For the appropriate clinical application of hypoxic cell sensitizers, the use of a ''vascularization index'', possibly determined by morphometric analysis of histological preparations, is suggested as a diagnostic parameter to characterize neoplasms besides current routine staging and grading of differentiation

  6. Determination of thorium in native gold by radiochemical neutron activation analysis

    International Nuclear Information System (INIS)

    Liu, Y.; Kraehenbuehl, U.

    1995-01-01

    Thorium concentrations in 11 native gold samples from different sources, e.g. placer gold, vein and lode gold were determined. Thorium was determined by radiochemical separation and measurement of protactinium from irradiated native gold samples. The chemical yield of the separation procedures is 90%. Other elements were measured by gamma-ray spectroscopy. The radiochemical separation procedures described in this work make accurate determination of Th concentrations in native gold at picogram concentrations possible. (orig.)

  7. ESOL facility for the generation and radiochemical separation of short half-life fission products

    International Nuclear Information System (INIS)

    Gehrke, R.J.; Meikrantz, D.H.; Baker, J.D.; Anderl, R.A.; Novick, V.J.; Greenwood, R.C.

    1988-01-01

    A facility has been developed at the Idaho National Engineering Laboratory (INEL) for the generation and rapid radiochemical separation of short half-life mixed fission products. This facility, referred to as the Idaho Elemental Separation On Line (ESOL), consists of electro-plated sources of spontaneously fissioning 252 Cf with a helium jet transport arrangement to continuously deliver short half-life, mixed fission products to the radiochemistry laboratory for rapid, computer controlled, radiochemical separations. 18 refs., 13 figs

  8. Current studies of biological materials using instrumental and radiochemical neutron activation analysis

    International Nuclear Information System (INIS)

    Fardy, J.J.; McOrist, G.D.; Farrar, Y.J.

    1985-01-01

    Instrumental neutron activation analysis still remains the preferred option when analysing the trace element distribution in a wide rage of materials by neutron activation analysis. However, when lower limits of detection are required or major interferences reduce the effectiveness of this technique, radiochemical neutron activation analysis is applied. This paper examines the current use of both methods and the development of rapid radiochemical techniques for analysis of the biological materials, hair, cow's milk, human's milk, milk powder, blood and blood serum

  9. A comparison of the radiochemical stability of different iodine-131 labelled metaiodobenzylguanidine formulations for therapeutic use

    International Nuclear Information System (INIS)

    Wafelman, A.R.; Beijnen, J.H.; Hoefnagel, C.A.; Maes, R.A.A.

    1994-01-01

    The results of a stability study of three commercially available formulations of [ 131 I]MIBG for therapeutic use and an unstabilized formulation, stored under various conditions, are presented. The stability was followed for 20 days. In all formulations tested, free [ 131 I]iodide, formed by radiolysis, was the most important radiochemical impurity. The pharmaceutical formulation with the largest amount of stabilizer was radiochemically - but not chemically -most stable. (author)

  10. Determination of radiochemical purity of 125I-TOC and 125I-F-PGA

    International Nuclear Information System (INIS)

    Yang Keya; Fan Wo; Zhang Youjiu; Xu Yujie; Zhu Ran; Hu Mingjiang

    2006-01-01

    To explore whether there is accordance among three determination methods of the radiochemical purity of [Tyr 3 ] octreotide (TOC) and folate-penicillin G amidase conjugate (F-PGA), which are both labeled with 125 I by Iodogen method, the RCP of the labelings are determined by high performance liquid chromatography (HPLC), paper chromatography and trichloroacetic acid (TCA) precipitation, in which four different concentrations of proteins are used to investigate the effect of them on the determination of RCP. It is shown that both HPLC and paper chromatography can separate the labelings from free iodine efficiently, though HPLC is the most precise and reliable method to determinate RCP of such labelings. In TCA precipitation, the RCP measured with 0.2%BSA is the lowest, but those with three other concentrations of the BSA are similar (P>0.05). When RCP 0.05), whereas higher than that with HPLC (P 10%, the RCP of 125 I-TOC obtained by TCA precipitation is a bit lower than those by two other methods (P 0.05), and there are no significant differences to determinate the RCP of 125 I-F-PGA (P>0.05). The three methods are correlated each other (r=0.0996-0.999, P<0.001). (authors)

  11. Crystallization and molecular-replacement studies of the monoclonal antibody mAbR310 specific for the (R)-HNE-modified protein

    International Nuclear Information System (INIS)

    Ito, Sohei; Tatsuda, Emi; Ishino, Kousuke; Suzuki, Kenichiro; Sakai, Hiroshi; Uchida, Koji

    2006-01-01

    Antigen-free Fab fragment of mAbR310, which recognizes (R)-HNE modified protein, has been crystallized. Initial phases have been obtained by molecular replacement. 4-Hydroxy-2-nonenal (HNE), a major racemic product of lipid peroxidation, reacts with histidine to form a stable HNE–histidine Michael addition-type adduct possessing three chiral centres in the cyclic hemiacetal structure. Monoclonal antibodies against HNE-modified protein have been widely used for assessing oxidative stress in vitro and in vivo. Here, the purification, crystallization and preliminary crystallographic analysis of a Fab fragment of novel monoclonal antibody R310 (mAbR310), which recognizes (R)-HNE-modified protein, are reported. The Fab fragment of mAbR310 was obtained by digestion with papain, purified and crystallized. Using hanging-drop vapour-diffusion crystallization techniques, crystals of mAbR310 Fab were obtained. The crystal belongs to the monoclinic space group C2 (unit-cell parameters a = 127.04, b = 65.31, c = 64.29 Å, β = 118.88°) and diffracted X-rays to a resolution of 1.84 Å. The asymmetric unit contains one molecule of mAbR310, with a corresponding crystal volume per protein weight of 2.51 Å 3 Da −1 and a solvent content of 51.0%

  12. X-ray crystallographic analysis of adipocyte fatty acid binding protein (aP2) modified with 4-hydroxy-2-nonenal

    Energy Technology Data Exchange (ETDEWEB)

    Hellberg, Kristina; Grimsrud, Paul A.; Kruse, Andrew C.; Banaszak, Leonard J.; Ohlendorf, Douglas H.; Bernlohr, David A. (UMM)

    2012-07-11

    Fatty acid binding proteins (FABP) have been characterized as facilitating the intracellular solubilization and transport of long-chain fatty acyl carboxylates via noncovalent interactions. More recent work has shown that the adipocyte FABP is also covalently modified in vivo on Cys117 with 4-hydroxy-2-nonenal (4-HNE), a bioactive aldehyde linked to oxidative stress and inflammation. To evaluate 4-HNE binding and modification, the crystal structures of adipocyte FABP covalently and noncovalently bound to 4-HNE have been solved to 1.9 {angstrom} and 2.3 {angstrom} resolution, respectively. While the 4-HNE in the noncovalently modified protein is coordinated similarly to a carboxylate of a fatty acid, the covalent form show a novel coordination through a water molecule at the polar end of the lipid. Other defining features between the two structures with 4-HNE and previously solved structures of the protein include a peptide flip between residues Ala36 and Lys37 and the rotation of the side chain of Phe57 into its closed conformation. Representing the first structure of an endogenous target protein covalently modified by 4-HNE, these results define a new class of in vivo ligands for FABPs and extend their physiological substrates to include bioactive aldehydes.

  13. Bioinformatics analysis to assess potential risks of allergenicity and toxicity of HRAP and PFLP proteins in genetically modified bananas resistant to Xanthomonas wilt disease.

    Science.gov (United States)

    Jin, Yuan; Goodman, Richard E; Tetteh, Afua O; Lu, Mei; Tripathi, Leena

    2017-11-01

    Banana Xanthomonas wilt (BXW) disease threatens banana production and food security throughout East Africa. Natural resistance is lacking among common cultivars. Genetically modified (GM) bananas resistant to BXW disease were developed by inserting the hypersensitive response-assisting protein (Hrap) or/and the plant ferredoxin-like protein (Pflp) gene(s) from sweet pepper (Capsicum annuum). Several of these GM banana events showed 100% resistance to BXW disease under field conditions in Uganda. The current study evaluated the potential allergenicity and toxicity of the expressed proteins HRAP and PFLP based on evaluation of published information on the history of safe use of the natural source of the proteins as well as established bioinformatics sequence comparison methods to known allergens (www.AllergenOnline.org and NCBI Protein) and toxins (NCBI Protein). The results did not identify potential risks of allergy and toxicity to either HRAP or PFLP proteins expressed in the GM bananas that might suggest potential health risks to humans. We recognize that additional tests including stability of these proteins in pepsin assay, nutrient analysis and possibly an acute rodent toxicity assay may be required by national regulatory authorities. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

  14. Dual-mode fluorophore-doped nickel nitrilotriacetic acid-modified silica nanoparticles combine histidine-tagged protein purification with site-specific fluorophore labeling.

    Science.gov (United States)

    Kim, Sung Hoon; Jeyakumar, M; Katzenellenbogen, John A

    2007-10-31

    We present the first example of a fluorophore-doped nickel chelate surface-modified silica nanoparticle that functions in a dual mode, combining histidine-tagged protein purification with site-specific fluorophore labeling. Tetramethylrhodamine (TMR)-doped silica nanoparticles, estimated to contain 700-900 TMRs per ca. 23 nm particle, were surface modified with nitrilotriacetic acid (NTA), producing TMR-SiO2-NTA-Ni2+. Silica-embedded TMR retains very high quantum yield, is resistant to quenching by buffer components, and is modestly quenched and only to a certain depth (ca. 2 nm) by surface-attached Ni2+. When exposed to a bacterial lysate containing estrogen receptor alpha ligand binding domain (ERalpha) as a minor component, these beads showed very high specificity binding, enabling protein purification in one step. The capacity and specificity of these beads for binding a his-tagged protein were characterized by electrophoresis, radiometric counting, and MALDI-TOF MS. ERalpha, bound to TMR-SiO2-NTA-Ni++ beads in a site-specific manner, exhibited good activity for ligand binding and for ligand-induced binding to coactivators in solution FRET experiments and protein microarray fluorometric and FRET assays. This dual-mode type TMR-SiO2-NTA-Ni2+ system represents a powerful combination of one-step histidine-tagged protein purification and site-specific labeling with multiple fluorophore species.

  15. Radiochemical analysis of concrete samples for decommission of nuclear reactors

    Energy Technology Data Exchange (ETDEWEB)

    Zapata-Garcia, Daniel; Wershofen, Herbert [Physikalisch-Technische Bundesanstalt (PTB), Bundesallee 100 38116, Braunschweig (Germany); Larijani, Cyrus; Sobrino-Petrirena, Maitane; Garcia-Miranda, Maria; Jerome, Simon M. [National Physical Laboratory (NPL), Hampton Road, Teddington, Middlesex, TW11 0LW (United Kingdom)

    2014-07-01

    Decommissioning of the oldest nuclear power reactors are some of the most challenging technological legacy issues many countries will face in forthcoming years, as many power reactors reach the end of their design lives. Decommissioning of nuclear reactors generates large amounts of waste that need to be classified according to their radioactive content. Approximately 10 % of the contaminated material ends up in different repositories (depending on their level of contamination) while the rest is decontaminated, measured and released into the environment or sent for recycling. Such classification needs to be done accurately in order to ensure that both the personnel involved in decommissioning and the population at large are not needlessly exposed to radiation or radioactive material and to minimise the environmental impact of such work. However, too conservative classification strategies should not be applied, in order to make proper use of radioactive waste repositories since space is limited and the full process must be cost-effective. Implicit in decommissioning and classification of waste is the need to analyse large amounts of material which usually combine a complex matrix with a non-homogeneous distribution of the radionuclides. Because the costs involved are large, it is possible to make great savings by the adoption of best available practices, such as the use of validated methods for on-site measurements and simultaneous determination of more than one radionuclide whenever possible. The work we present deals with the development and the validation of a procedure for the simultaneous determination of {sup 241}Am, plutonium isotopes, uranium isotopes and {sup 90}Sr in concrete samples. Samples are firstly ground and fused with LiBO{sub 2} and Li{sub 2}B{sub 4}O{sub 7}. After dissolution of the fused sample, silicate and alkaline elements are removed followed by radiochemical separation of the target radionuclides using extraction chromatography. Measurement

  16. Characterization of the single transmembrane domain of human receptor activity-modifying protein 3 in adrenomedullin receptor internalization

    International Nuclear Information System (INIS)

    Kuwasako, Kenji; Kitamura, Kazuo; Nagata, Sayaka; Nozaki, Naomi; Kato, Johji

    2012-01-01

    Highlights: ► RAMP3 mediates CLR internalization much less effectively than does RAMP2. ► The RAMP3 TMD participates in the negative regulation of CLR/RAMP3 internalization. ► A new strategy of promoting internalization and resensitization of the receptor was found. -- Abstract: Two receptor activity-modifying proteins (RAMP2 and RAMP3) enable calcitonin receptor-like receptor (CLR) to function as two heterodimeric receptors (CLR/RAMP2 and CLR/RAMP3) for adrenomedullin (AM), a potent cardiovascular protective peptide. Following AM stimulation, both receptors undergo rapid internalization through a clathrin-dependent pathway, after which CLR/RAMP3, but not CLR/RAMP2, can be recycled to the cell surface for resensitization. However, human (h)RAMP3 mediates CLR internalization much less efficiently than does hRAMP2. Therefore, the molecular basis of the single transmembrane domain (TMD) and the intracellular domain of hRAMP3 during AM receptor internalization was investigated by transiently transfecting various RAMP chimeras and mutants into HEK-293 cells stably expressing hCLR. Flow cytometric analysis revealed that substituting the RAMP3 TMD with that of RAMP2 markedly enhanced AM-induced internalization of CLR. However, this replacement did not enhance the cell surface expression of CLR, [ 125 I]AM binding affinity or AM-induced cAMP response. More detailed analyses showed that substituting the Thr 130 –Val 131 sequence in the RAMP3 TMD with the corresponding sequence (Ile 157 –Pro 158 ) from RAMP2 significantly enhanced AM-mediated CLR internalization. In contrast, substituting the RAMP3 target sequence with Ala 130 –Ala 131 did not significantly affect CLR internalization. Thus, the RAMP3 TMD participates in the negative regulation of CLR/RAMP3 internalization, and the aforementioned introduction of the Ile–Pro sequence into the RAMP3 TMD may be a strategy for promoting receptor internalization/resensitization.

  17. Comparison of different thin layer detection techniques to determine the radiochemical purity of radiopharmaceuticals

    International Nuclear Information System (INIS)

    Hammermaier, A.; Reich, E.; Boegl, W.

    1985-01-01

    Ten radiopharmaceuticals frequently used in clinical treatment were examined as to their radiochemical purity by paper and thin layer chromatography or electrophoresis, respectively. It is known that radiochemical impurities may result in an unnecessary exposure of the patients to be examined. Other than determining the radiochemical purity of several radiopharmaceuticals, a comparison of the different measuring methods of distributing activity on radiochromatograms or electropherograms is intended by this study. For this, the activity distribution in the developed radiochromatograms was assessed by four different measuring methods (TLC-linear analyzer, TLC-scanner with NaI(Tl) detector, TLC-scanner with gas flow counter and NaI(Tl) well-typ counter). As shown by the above analysis, only the TLC-linear analyzer and the NaI(Tl) well-typ counter (measurement of chromatograms or electropherograms cut into strips) are generally suitable methods for determining the radiochemical purity of radiochemicals, the TLC-scanner with gas flow counter is usable in most cases, while TLC-scanner with NaI(Tl) detector is yielding unsatisfactory results. (orig.) [de

  18. Lack of detectable allergenicity in genetically modified maize containing "Cry" proteins as compared to native maize based on in silico & in vitro analysis.

    Directory of Open Access Journals (Sweden)

    Chandni Mathur

    Full Text Available Genetically modified, (GM crops with potential allergens must be evaluated for safety and endogenous IgE binding pattern compared to native variety, prior to market release.To compare endogenous IgE binding proteins of three GM maize seeds containing Cry 1Ab,1Ac,1C transgenic proteins with non GM maize.An integrated approach of in silico & in vitro methods was employed. Cry proteins were tested for presence of allergen sequence by FASTA in allergen databases. Biochemical assays for maize extracts were performed. Specific IgE (sIgE and Immunoblot using food sensitized patients sera (n = 39 to non GM and GM maize antigens was performed.In silico approaches, confirmed for non sequence similarity of stated transgenic proteins in allergen databases. An insignificant (p> 0.05 variation in protein content between GM and non GM maize was observed. Simulated Gastric Fluid (SGF revealed reduced number of stable protein fractions in GM then non GM maize which might be due to shift of constituent protein expression. Specific IgE values from patients showed insignificant difference in non GM and GM maize extracts. Five maize sensitized cases, recognized same 7 protein fractions of 88-28 kD as IgE bindng in both GM and non-GM maize, signifying absence of variation. Four of the reported IgE binding proteins were also found to be stable by SGF.Cry proteins did not indicate any significant similarity of >35% in allergen databases. Immunoassays also did not identify appreciable differences in endogenous IgE binding in GM and non GM maize.

  19. Lack of Detectable Allergenicity in Genetically Modified Maize Containing “Cry” Proteins as Compared to Native Maize Based on In Silico & In Vitro Analysis

    Science.gov (United States)

    Mathur, Chandni; Kathuria, Pooran C.; Dahiya, Pushpa; Singh, Anand B.

    2015-01-01

    Background Genetically modified, (GM) crops with potential allergens must be evaluated for safety and endogenous IgE binding pattern compared to native variety, prior to market release. Objective To compare endogenous IgE binding proteins of three GM maize seeds containing Cry 1Ab,1Ac,1C transgenic proteins with non GM maize. Methods An integrated approach of in silico & in vitro methods was employed. Cry proteins were tested for presence of allergen sequence by FASTA in allergen databases. Biochemical assays for maize extracts were performed. Specific IgE (sIgE) and Immunoblot using food sensitized patients sera (n = 39) to non GM and GM maize antigens was performed. Results In silico approaches, confirmed for non sequence similarity of stated transgenic proteins in allergen databases. An insignificant (p> 0.05) variation in protein content between GM and non GM maize was observed. Simulated Gastric Fluid (SGF) revealed reduced number of stable protein fractions in GM then non GM maize which might be due to shift of constituent protein expression. Specific IgE values from patients showed insignificant difference in non GM and GM maize extracts. Five maize sensitized cases, recognized same 7 protein fractions of 88-28 kD as IgE bindng in both GM and non-GM maize, signifying absence of variation. Four of the reported IgE binding proteins were also found to be stable by SGF. Conclusion Cry proteins did not indicate any significant similarity of >35% in allergen databases. Immunoassays also did not identify appreciable differences in endogenous IgE binding in GM and non GM maize. PMID:25706412

  20. Identification and modulation of the key amino acid residue responsible for the pH sensitivity of neoculin, a taste-modifying protein.

    Directory of Open Access Journals (Sweden)

    Ken-ichiro Nakajima

    Full Text Available Neoculin occurring in the tropical fruit of Curculigo latifolia is currently the only protein that possesses both a sweet taste and a taste-modifying activity of converting sourness into sweetness. Structurally, this protein is a heterodimer consisting of a neoculin acidic subunit (NAS and a neoculin basic subunit (NBS. Recently, we found that a neoculin variant in which all five histidine residues are replaced with alanine elicits intense sweetness at both neutral and acidic pH but has no taste-modifying activity. To identify the critical histidine residue(s responsible for this activity, we produced a series of His-to-Ala neoculin variants and evaluated their sweetness levels using cell-based calcium imaging and a human sensory test. Our results suggest that NBS His11 functions as a primary pH sensor for neoculin to elicit taste modification. Neoculin variants with substitutions other than His-to-Ala were further analyzed to clarify the role of the NBS position 11 in the taste-modifying activity. We found that the aromatic character of the amino acid side chain is necessary to elicit the pH-dependent sweetness. Interestingly, since the His-to-Tyr variant is a novel taste-modifying protein with alternative pH sensitivity, the position 11 in NBS can be critical to modulate the pH-dependent activity of neoculin. These findings are important for understanding the pH-sensitive functional changes in proteinaceous ligands in general and the interaction of taste receptor-taste substance in particular.

  1. Method of determination of radiochemical purity of gallium-67 citrate injection

    International Nuclear Information System (INIS)

    Wang Quanji

    1985-01-01

    A simple method is used to compare the effect of five developing agents on the radiochemical purity of neutral products of 67 GaCit and on Rsub(f) values. Two preferable developing agents are recommended as suitable for the identification of 67 GaCit injection in its production. The effect of six pH values of different developing agents on radiochemical purity, Rsub(f) and chromatogram are compared for the neutral products. The results of the experiments show that the ascending paper chromatography with 1:2:4 pyridine/ethanol/water and 85:15 methanol/water is preferable for the determination of the radiochemical purity of 67 GaCit. The other developing agents also can be used if there are not any impurities except gallium radioisotopes

  2. Northern Marshall Islands Radiological Survey: a quality-control program for a radiochemical analyses

    International Nuclear Information System (INIS)

    Jennings, C.D.; Mount, M.E.

    1983-08-01

    More than 16,000 radiochemical analyses were performed on about 5400 samples of soils, vegetation, animals, fish, invertebrates, and water to establish amounts of 90 Sr, 137 Cs, 241 Am, and plutonium isotopes in the Northern Marshall Islands. Three laboratories were contracted by Lawrence Livermore National Laboratory to perform the radiochemical analyses: Environmental Analysis Laboratory (EAL), Richmond, California; Eberline Instrument Corporation (EIC), Albuquerque, New Mexico; and Laboratory of Radiation Ecology (LRE), University of Washington, Seattle, Washington. The analytical precision and accuracy were monitored by regularly including duplicate samples and natural matrix standards in each group of about 100 samples analyzed. Based on the duplicates and standards, over 83% of the radiochemical analyses in this survey were acceptable - 97% of the analyses by EAL, 45% of the analyses by EIC, and 98% of the analyses by LRE

  3. High-protein diet modifies colonic microbiota and luminal environment but not colonocyte metabolism in the rat model: the increased luminal bulk connection.

    Science.gov (United States)

    Liu, Xinxin; Blouin, Jean-Marc; Santacruz, Arlette; Lan, Annaïg; Andriamihaja, Mireille; Wilkanowicz, Sabina; Benetti, Pierre-Henri; Tomé, Daniel; Sanz, Yolanda; Blachier, François; Davila, Anne-Marie

    2014-08-15

    High-protein diets are used for body weight reduction, but consequences on the large intestine ecosystem are poorly known. Here, rats were fed for 15 days with either a normoproteic diet (NP, 14% protein) or a hyperproteic-hypoglucidic isocaloric diet (HP, 53% protein). Cecum and colon were recovered for analysis. Short- and branched-chain fatty acids, as well as lactate, succinate, formate, and ethanol contents, were markedly increased in the colonic luminal contents of HP rats (P diet, whereas the amount of butyrate in feces was increased (P diet consumption allows maintenance in the luminal butyrate concentration and thus its metabolism in colonocytes despite modified microbiota composition and increased substrate availability. Copyright © 2014 the American Physiological Society.

  4. Radiolabeling, quality control and radiochemical purity assessment of 99mTc-HYNIC-TOC

    International Nuclear Information System (INIS)

    Melero, Laura T.U.H.; Araujo, Elaine B.; Mengatti, Jair

    2009-01-01

    Somatostatine receptors are widely expressed by several tumors, especially of the neuroendocrine origin. In vivo images of these tumors using radiolabeled somatostatine analogues became a useful clinical tool in oncology. The aim of this work was the radiolabeling of the somatostatine analogue HYNIC-TOC with 99mTc as well as the evaluation of the radiochemical stability and quality control of labeled complex. 99mTc-HYNIC-TOC was produced by labeling conditions using 20 μg of peptide, 20 mg of tricine and 10 mg of EDDA as coligands, 1110 MBq of 99mTc (99Mo-99mTc IPEN-TEC generator) and 15 μg of SnCl 2 .2H 2 O. The reaction proceeds for 10 minutes at boiling water bath. Radiochemical purity of labeled preparation was evaluated by different chromatographic systems: ITLC-SG in methanol:ammonium acetate (1:1); TLC-SG in sodium citrate buffer 0.1 N pH 5.0 and methylethylketone, and HPLC employing column C-18, 5 μm, 4.6 mm x 250 mm, UV (220 nm), radioactivity detectors, 1 mL/minute flow of acetonitrile and trifluoroacetic acid solution 0.1 %. Labeled compound has been found radiochemically stable for 5 hours and radiochemical purity was higher than 90 %. The thin layer chromatographic systems enabled the separation of radiochemical species presented in the labeled mixture as well as HPLC system. The labeling procedure studied resulted in high radiochemical yield and easy preparation. Future works include the preparation of a lyophilized reagent to make feasible the preparation of 99mTc-HYNIC-TOC at nuclear medicine services in order to study the clinical potential of the radiopharmaceutical in diagnostic and staging of neuroendocrine tumors. (author)

  5. Radiolabeling, quality control and radiochemical purity assessment of {sup 99m}Tc-HYNIC-TOC

    Energy Technology Data Exchange (ETDEWEB)

    Melero, Laura T.U.H.; Araujo, Elaine B.; Mengatti, Jair [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil)

    2009-07-01

    Somatostatine receptors are widely expressed by several tumors, especially of the neuroendocrine origin. In vivo images of these tumors using radiolabeled somatostatine analogues became a useful clinical tool in oncology. The aim of this work was the radiolabeling of the somatostatine analogue HYNIC-TOC with 99mTc as well as the evaluation of the radiochemical stability and quality control of labeled complex. 99mTc-HYNIC-TOC was produced by labeling conditions using 20 {mu}g of peptide, 20 mg of tricine and 10 mg of EDDA as coligands, 1110 MBq of 99mTc (99Mo-99mTc IPEN-TEC generator) and 15 {mu}g of SnCl{sub 2}.2H{sub 2}O. The reaction proceeds for 10 minutes at boiling water bath. Radiochemical purity of labeled preparation was evaluated by different chromatographic systems: ITLC-SG in methanol:ammonium acetate (1:1); TLC-SG in sodium citrate buffer 0.1 N pH 5.0 and methylethylketone, and HPLC employing column C-18, 5 {mu}m, 4.6 mm x 250 mm, UV (220 nm), radioactivity detectors, 1 mL/minute flow of acetonitrile and trifluoroacetic acid solution 0.1 %. Labeled compound has been found radiochemically stable for 5 hours and radiochemical purity was higher than 90 %. The thin layer chromatographic systems enabled the separation of radiochemical species presented in the labeled mixture as well as HPLC system. The labeling procedure studied resulted in high radiochemical yield and easy preparation. Future works include the preparation of a lyophilized reagent to make feasible the preparation of 99mTc-HYNIC-TOC at nuclear medicine services in order to study the clinical potential of the radiopharmaceutical in diagnostic and staging of neuroendocrine tumors. (author)

  6. Identification of Atg2 and ArfGAP1 as Candidate Genetic Modifiers of the Eye Pigmentation Phenotype of Adaptor Protein-3 (AP-3) Mutants in Drosophila melanogaster.

    Science.gov (United States)

    Rodriguez-Fernandez, Imilce A; Dell'Angelica, Esteban C

    2015-01-01

    The Adaptor Protein (AP)-3 complex is an evolutionary conserved, molecular sorting device that mediates the intracellular trafficking of proteins to lysosomes and related organelles. Genetic defects in AP-3 subunits lead to impaired biogenesis of lysosome-related organelles (LROs) such as mammalian melanosomes and insect eye pigment granules. In this work, we have performed a forward screening for genetic modifiers of AP-3 function in the fruit fly, Drosophila melanogaster. Specifically, we have tested collections of large multi-gene deletions--which together covered most of the autosomal chromosomes-to identify chromosomal regions that, when deleted in single copy, enhanced or ameliorated the eye pigmentation phenotype of two independent AP-3 subunit mutants. Fine-mapping led us to define two non-overlapping, relatively small critical regions within fly chromosome 3. The first critical region included the Atg2 gene, which encodes a conserved protein involved in autophagy. Loss of one functional copy of Atg2 ameliorated the pigmentation defects of mutants in AP-3 subunits as well as in two other genes previously implicated in LRO biogenesis, namely Blos1 and lightoid, and even increased the eye pigment content of wild-type flies. The second critical region included the ArfGAP1 gene, which encodes a conserved GTPase-activating protein with specificity towards GTPases of the Arf family. Loss of a single functional copy of the ArfGAP1 gene ameliorated the pigmentation phenotype of AP-3 mutants but did not to modify the eye pigmentation of wild-type flies or mutants in Blos1 or lightoid. Strikingly, loss of the second functional copy of the gene did not modify the phenotype of AP-3 mutants any further but elicited early lethality in males and abnormal eye morphology when combined with mutations in Blos1 and lightoid, respectively. These results provide genetic evidence for new functional links connecting the machinery for biogenesis of LROs with molecules implicated in

  7. Use of reference materials for quality control of elemental analysis by neutron activation with radiochemical separation

    International Nuclear Information System (INIS)

    Woittiez, J.R.W.

    1990-01-01

    This paper describes the use of certified reference materials to monitor the long-term quality of radiochemical separations. The practical limitations which determine the actual design of the quality control are discussed. The hypothesis that the high yield of the radiochemical separation will be constant with time has been checked and validated for the elements Zn, Fe, Co, Cd, Mo and to a lesser extent for W and Th using NBS SRM 1577A, BCR CRM 274 and IAEA RM A-11. This validation could not be made for the elements Cr, Au, and Ag. Especially for Cr there is a serious lack of appropiate certified reference materials. (orig.)

  8. Radiochemical procedures for determination of selected members of the uranium and thorium series

    International Nuclear Information System (INIS)

    Smithson, G.L.

    1979-01-01

    The radiochemical procedures contained in this manual are adaptations of those developed and published by many radiochemists. In many cases the identity of the originator is not clear and usually modifications in the original procedure have been made by subsequent workers. Nearly all of the basic radiochemical techniques and separations in use today were developed during the Manhattan Project and can be found in U.S.A.E.C. reports published from 1945 to 1953. This manual contains methods for the determination of Pb-210, Po-210; Ra-226, Ra-228, Th-228, Th-230 and Th-232. (auth)

  9. Critical evaluation of the determination of zirconium and hafnium by instrumental and radiochemical neutron activation analysis

    International Nuclear Information System (INIS)

    Burger, Mario; Kraehenbuehl, Urs

    1991-01-01

    Neutron activation analysis (instrument or radiochemical) is suitable for the determination of zirconium and hafnium in samples of geochemical origin only when sufficient attention is paid to inter-fering nuclides. The size of the necessary correction for INAA depends on the composition of the sample; this problem is discussed. The radio-chemical technique which is recommended involves separation of the samples, precipitations and anion-exchange separation. Results are given for various standard reference materials and for meteorites. (author). 12 refs.; 1 fig.; 9 tabs

  10. On the methodology of radiochemical neutron activation analysis of noble metals

    International Nuclear Information System (INIS)

    Chai, C.F.; Ma, S.L.; Mao, X.Y.; Liao, K.N.; Liu, W.C.

    1986-01-01

    Two different radiochemical procedures were developed: chelate ion resin exchange and amine solvent extraction. Two kinds of new Chinese chelate resins (NANKAI-3926 and BEI-5) and a new long-chain primary amine N 1923 were compared with Srafion NMRR and the tertiary amine N 235 in absorption performance of noble metals, respectively. Influences of various experimental conditions, e.g. sample digestion, acidity, equilibrium time, as well as elution of noble metals, on analytical sensitivity and chemical yield were discussed. Combining with neutron activation, the radiochemical separation procedures developed were used to determine the noble metal contents in the geological samples from Permina/Triassic boundary in South China. (author)

  11. Abyssal sediment erosion from the Central Indian Basin: Evidence from radiochemical and radiolarian studies

    Digital Repository Service at National Institute of Oceanography (India)

    Banakar, V.K.; Gupta, S.M.; Padmavati, V.K.

    ) 167-173 167 Elsevier Science Publishers B.V., Amsterdam Letter Section Abyssal sediment erosion in the Central Indian Basin: Evidence from radiochemical and radiolarian studies V.K. Banakar, S.M. Gupta and V.K. Padmavathi National Institute... of Oceanography, Dona-Paula, Goa-403 004, India (Revision accepted September 17, 1990) ABSTRACT Banakar, V.K., Gupta, S.M. and Padmavathi, V.K., 1991. Abyssal sediment erosion from the Central Indian Basin: Evi- dence from radiochemical and radiolarian studies...

  12. Use of ethyl-α-isonitrosoacetoacetate in the rapid estimation and radiochemical separation of gold

    International Nuclear Information System (INIS)

    Sawant, A.D.; Haldar, B.C.

    1978-01-01

    The use of ethyl-α-isonitrosoacetoacetate in the rapid estimation and radiochemical separation of gold is reported. As low as 5.00 mg of Au can be estimated with an accuracy better than 1%. Decontamination values against platinum metals and other metals usually associated with Au are greater than 10 5 . Isotopes and results are tabulated. The time required for radiochemical separation is around 20 min and the recovery of Au is better than 80%. γ-activities were measured with a single channel analyser and NaI(Tl) detector. β-activities were counted on a thin end-window type GM counter. (T.I.)

  13. Kit preparation of 153Sm-EDTMP and factors affecting radiochemical purity and stability

    International Nuclear Information System (INIS)

    Ferro-Flores, G.; Tendilla, J.I.; Lopez-Gomez, M.A.; Aguilar-Hernandez, F.; Gonzalez-Zavala, M.A.; Parades-Gutierrez, L.; Avila-Ramirez, E.

    1996-01-01

    A fast kit method was developed for the production of 153 Sm-EDTMP in two steps avoiding the use of nitric acid, evaporation and sterilization of the final solution by autoclave. Methods of analysis for the determination of chemical and radiochemical purity in the radiopharmaceutical solution were established. Factors affecting radiochemical purity and stability of the complex as the molar ratio of EDTMP/Sm, concentration of phosphate buffer and neutralization of EDTMP prior kit preparation were also analyzed. The use of this radiopharmaceutical in rabbits and patients showed selective skeletal uptake. (author). 5 refs., 4 figs., 3 tabs

  14. The role of high performance liquid chromatography in radiochemical/radiopharmaceutical synthesis and quality assurance

    International Nuclear Information System (INIS)

    Boothe, T.E.; Emran, A.M.

    1990-01-01

    The usefulness of HPLC in all areas of radiopharmaceutics has been demonstrated in numerous laboratories, particularly in the development of in-house radiopharmaceuticals for SPECT and PET. HPLC continues to be a powerful tool in preparation and quality assurance (QA) as illustrated in such areas as chemical and radiochemical identification; product separation and isolation; preparative scale purification; and specific activity determination. A review of established HPLC techniques in radiopharmaceutics will be presented. Examples from the literature as well as newer applications will be used in an attempt to assess and define the present-day role of HPLC in the preparation of radiochemicals and radiopharmaceuticals with emphasis on QA

  15. Radiochemical methodologies applied to analytical characterization of low and intermediate level wastes from nuclear power plants

    International Nuclear Information System (INIS)

    Monteiro, Roberto Pellacani G.; Júnior, Aluísio Souza R.; Kastner, Geraldo F.; Temba, Eliane S.C.; Oliveira, Thiago C. de; Amaral, Ângela M.; Franco, Milton B.

    2017-01-01

    The aim of this work is to present radiochemical methodologies developed at CDTN/CNEN in order to answer a program for isotopic inventory of radioactive wastes from Brazilian Nuclear Power Plants. In this program some radionuclides, 3 H, 14 C, 55 Fe, 59 Ni, 63 Ni, 90 Sr, 93 Zr, 94 Nb, 99 Tc, 129 I, 235 U, 238 U, 238 Pu, 239 + 240 Pu, 241 Pu, 242 Pu, 241 Am, 242 Cm e 243 + 244 Cm, were determined in Low Level Wastes (LLW) and Intermediate Level Wastes (ILW) and a protocol of analytical methodologies based on radiochemical separation steps and spectrometric and nuclear techniques was established. (author)

  16. Senp1 Is Essential for Desumoylating Sumo1-Modified Proteins but Dispensable for Sumo2 and Sumo3 Deconjugation in the Mouse Embryo

    Directory of Open Access Journals (Sweden)

    Prashant Sharma

    2013-05-01

    Full Text Available Posttranslational modification with small ubiquitin-like modifier (Sumo regulates numerous cellular and developmental processes. Sumoylation is dynamic with deconjugation by Sumo-specific proteases (Senps regulating steady-state levels. Different Senps are found in distinct subcellular domains, which may limit their deconjugation activity to colocalizing Sumo-modified proteins. In vitro, Senps can discriminate between the different Sumo paralogs: Sumo1 versus the highly related Sumo2 and Sumo3 (Sumo2/3, which can form poly-Sumo chains. However, a full understanding of Senp specificity in vivo is still lacking. Here, using biochemical and genetic approaches, we establish that Senp1 has an essential, nonredundant function to desumoylate Sumo1-modified proteins during mouse embryonic development. Senp1 specificity for Sumo1 conjugates represents an intrinsic function and not simply a product of colocalization. In contrast, Senp1 has only a limited role in Sumo2/3 desumoylation, although it may regulate Sumo1-mediated termination of poly-Sumo2/3 chains.

  17. Roles of Trm9- and ALKBH8-like proteins in the formation of modified wobble uridines in Arabidopsis tRNA

    DEFF Research Database (Denmark)

    Leihne, Vibeke; Kirpekar, Finn; Vågbø, Cathrine B

    2011-01-01

    Uridine at the wobble position of tRNA is usually modified, and modification is required for accurate and efficient protein translation. In eukaryotes, wobble uridines are modified into 5-methoxycarbonylmethyluridine (mcm(5)U), 5-carbamoylmethyluridine (ncm(5)U) or derivatives thereof. Here, we...... demonstrate, both by in vitro and in vivo studies, that the Arabidopsis thaliana methyltransferase AT1G31600, denoted by us AtTRM9, is responsible for the final step in mcm(5)U formation, thus representing a functional homologue of the Saccharomyces cerevisiae Trm9 protein. We also show that the enzymatic...... activity of AtTRM9 depends on either one of two closely related proteins, AtTRM112a and AtTRM112b. Moreover, we demonstrate that AT1G36310, denoted AtALKBH8, is required for hydroxylation of mcm(5)U to (S)-mchm(5)U in tRNA(Gly)(UCC), and has a function similar to the mammalian dioxygenase ALKBH8...

  18. Protein-energy malnutrition induces an aberrant acute-phase response and modifies the circadian rhythm of core temperature.

    Science.gov (United States)

    Smith, Shari E; Ramos, Rafaela Andrade; Refinetti, Roberto; Farthing, Jonathan P; Paterson, Phyllis G

    2013-08-01

    Protein-energy malnutrition (PEM), present in 12%-19% of stroke patients upon hospital admission, appears to be a detrimental comorbidity factor that impairs functional outcome, but the mechanisms are not fully elucidated. Because ischemic brain injury is highly temperature-sensitive, the objectives of this study were to investigate whether PEM causes sustained changes in temperature that are associated with an inflammatory response. Activity levels were recorded as a possible explanation for the immediate elevation in temperature upon introduction to a low protein diet. Male, Sprague-Dawley rats (7 weeks old) were fed a control diet (18% protein) or a low protein diet (PEM, 2% protein) for either 7 or 28 days. Continuous core temperature recordings from bioelectrical sensor transmitters demonstrated a rapid increase in temperature amplitude, sustained over 28 days, in response to a low protein diet. Daily mean temperature rose transiently by day 2 (p = 0.01), falling to normal by day 4 (p = 0.08), after which mean temperature continually declined as malnutrition progressed. There were no alterations in activity mean (p = 0.3) or amplitude (p = 0.2) that were associated with the early rise in mean temperature. Increased serum alpha-2-macroglobulin (p protein diet had no effect on the signaling pathway of the pro-inflammatory transcription factor, NFκB, in the hippocampus. In conclusion, PEM induces an aberrant and sustained acute-phase response coupled with long-lasting effects on body temperature.

  19. Protein sequence analysis by incorporating modified chaos game and physicochemical properties into Chou's general pseudo amino acid composition.

    Science.gov (United States)

    Xu, Chunrui; Sun, Dandan; Liu, Shenghui; Zhang, Yusen

    2016-10-07

    In this contribution we introduced a novel graphical method to compare protein sequences. By mapping a protein sequence into 3D space based on codons and physicochemical properties of 20 amino acids, we are able to get a unique P-vector from the 3D curve. This approach is consistent with wobble theory of amino acids. We compute the distance between sequences by their P-vectors to measure similarities/dissimilarities among protein sequences. Finally, we use our method to analyze four datasets and get better results compared with previous approaches. Copyright © 2016 Elsevier Ltd. All rights reserved.

  20. Extracellular vesicles shed by melanoma cells contain a modified form of H1.0 linker histone and H1.0 mRNA-binding proteins.

    Science.gov (United States)

    Schiera, Gabriella; Di Liegro, Carlo Maria; Puleo, Veronica; Colletta, Oriana; Fricano, Anna; Cancemi, Patrizia; Di Cara, Gianluca; Di Liegro, Italia

    2016-11-01

    Extracellular vesicles (EVs) are now recognized as a fundamental way for cell-to-cell horizontal transfer of properties, in both physiological and pathological conditions. Most of EV-mediated cross-talk among cells depend on the exchange of proteins, and nucleic acids, among which mRNAs, and non-coding RNAs such as different species of miRNAs. Cancer cells, in particular, use EVs to discard molecules which could be dangerous to them (for example differentiation-inducing proteins such as histone H1.0, or antitumor drugs), to transfer molecules which, after entering the surrounding cells, are able to transform their phenotype, and even to secrete factors, which allow escaping from immune surveillance. Herein we report that melanoma cells not only secrete EVs which contain a modified form of H1.0 histone, but also transport the corresponding mRNA. Given the already known role in tumorigenesis of some RNA binding proteins (RBPs), we also searched for proteins of this class in EVs. This study revealed the presence in A375 melanoma cells of at least three RBPs, with apparent MW of about 65, 45 and 38 kDa, which are able to bind H1.0 mRNA. Moreover, we purified one of these proteins, which by MALDI-TOF mass spectrometry was identified as the already known transcription factor MYEF2.

  1. Low-molecular weight protein profiling of genetically modified maize using fast liquid chromatography electrospray ionization and time-of-flight mass spectrometry.

    Science.gov (United States)

    Koc, Anna; Cañuelo, Ana; Garcia-Reyes, Juan F; Molina-Diaz, Antonio; Trojanowicz, Marek

    2012-06-01

    In this work, the use of liquid chromatography coupled to electrospray time-of-flight mass spectrometry (LC-TOFMS) has been evaluated for the profiling of relatively low-molecular weight protein species in both genetically modified (GM) and non-GM maize. The proposed approach consisted of a straightforward sample fractionation with different water and ethanol-based buffer solutions followed by separation and detection of the protein species using liquid chromatography with a small particle size (1.8 μm) C(18) column and electrospray-time-of-flight mass spectrometry detection in the positive ionization mode. The fractionation of maize reference material containing different content of transgenic material (from 0 to 5% GM) led to five different fractions (albumins, globulins, zeins, zein-like glutelins, and glutelins), all of them containing different protein species (from 2 to 52 different species in each fraction). Some relevant differences in the quantity and types of protein species were observed in the different fractions of the reference material (with different GM contents) tested, thus revealing the potential use of the proposed approach for fast protein profiling and to detect tentative GMO markers in maize. © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  2. Rapid Radiochemical Analyses in Support of Fukushima Nuclear Accident - 13196

    Energy Technology Data Exchange (ETDEWEB)

    Maxwell, Sherrod L.; Culligan, Brian K.; Hutchison, Jay B. [Savannah River National Laboratory, Building 735-B, Aiken, SC 29808 (United States)

    2013-07-01

    discussed. Air filter samples were reported within twenty-four (24) hours of receipt using rapid techniques published previously. [11] The rapid reporting of high quality analytical data arranged through the U.S. Department of Energy Consequence Management Home Team was critical to allow the government of Japan to readily evaluate radiological impacts from the nuclear reactor incident to both personnel and the environment. SRNL employed unique rapid methods capability for radionuclides to support Japan that can also be applied to environmental, bioassay and waste management samples. New rapid radiochemical techniques for radionuclides in soil and other environmental matrices as well as some of the unique challenges associated with this work will be presented that can be used for application to environmental monitoring, environmental remediation, decommissioning and decontamination activities. (authors)

  3. An in vitro tag-and-modify protein sample generation method for single-molecule fluorescence resonance energy transfer.

    Science.gov (United States)

    Hamadani, Kambiz M; Howe, Jesse; Jensen, Madeleine K; Wu, Peng; Cate, Jamie H D; Marqusee, Susan

    2017-09-22

    Biomolecular systems exhibit many dynamic and biologically relevant properties, such as conformational fluctuations, multistep catalysis, transient interactions, folding, and allosteric structural transitions. These properties are challenging to detect and engineer using standard ensemble-based techniques. To address this drawback, single-molecule methods offer a way to access conformational distributions, transient states, and asynchronous dynamics inaccessible to these standard techniques. Fluorescence-based single-molecule approaches are parallelizable and compatible with multiplexed detection; to date, however, they have remained limited to serial screens of small protein libraries. This stems from the current absence of methods for generating either individual dual-labeled protein samples at high throughputs or protein libraries compatible with multiplexed screening platforms. Here, we demonstrate that by combining purified and reconstituted in vitro translation, quantitative unnatural amino acid incorporation via AUG codon reassignment, and copper-catalyzed azide-alkyne cycloaddition, we can overcome these challenges for target proteins that are, or can be, methionine-depleted. We present an in vitro parallelizable approach that does not require laborious target-specific purification to generate dual-labeled proteins and ribosome-nascent chain libraries suitable for single-molecule FRET-based conformational phenotyping. We demonstrate the power of this approach by tracking the effects of mutations, C-terminal extensions, and ribosomal tethering on the structure and stability of three protein model systems: barnase, spectrin, and T4 lysozyme. Importantly, dual-labeled ribosome-nascent chain libraries enable single-molecule co-localization of genotypes with phenotypes, are well suited for multiplexed single-molecule screening of protein libraries, and should enable the in vitro directed evolution of proteins with designer single-molecule conformational

  4. Titania nanotube-modified screen printed carbon electrodes enhance the sensitivity in the electrochemical detection of proteins.

    Science.gov (United States)

    Mandal, Soumit S; Navratna, Vikas; Sharma, Pratyush; Gopal, B; Bhattacharyya, Aninda J

    2014-08-01

    The use of titania nanotubes (TiO2-NT) as the working electrode provides a substantial improvement in the electrochemical detection of proteins. A biosensor designed using this strategy provided a robust method to detect protein samples at very low concentrations (Cprotein ca 1ng/μl). Reproducible measurements on protein samples at this concentration (Ip,a of 80+1.2μA) could be achieved using a sample volume of ca 30μl. We demonstrate the feasibility of this strategy for the accurate detection of penicillin binding protein, PBP2a, a marker for methicillin resistant Staphylococcus aureus (MRSA). The selectivity and efficiency of this sensor were also validated using other diverse protein preparations such as a recombinant protein tyrosine phosphatase (PTP10D) and bovine serum albumin (BSA). This electrochemical method also presents a substantial improvement in the time taken (few minutes) when compared to conventional enzyme-linked immunosorbent assay (ELISA) protocols. It is envisaged that this sensor could substantially aid in the rapid diagnosis of bacterial infections in resource strapped environments. Copyright © 2014 Elsevier B.V. All rights reserved.

  5. Au nanocrystals grown on a better-defined one-dimensional tobacco mosaic virus coated protein template genetically modified by a hexahistidine tag

    International Nuclear Information System (INIS)

    Liu Nan; Zhang Wei; Luo Zhaopeng; Zhai Niu; Zhang Hongfei; Li Zhonghao; Jiang Xingyi; Tang Gangling; Hu Qingyuan; Wang Chong; Tian Dandan

    2012-01-01

    In this paper, tobacco mosaic virus (TMV) coated protein (CP) was genetically modified by introducing a hexahistidine tag into it for a well-defined one-dimensional template, on which Au nanocrystals (NCs) were grown. The results showed that genetic modification could not only ameliorate the one-dimensional structure of the template, but also improve the growth density of Au NCs on the template. This indicated that genetic modification could be an effective method to modulate the structure of the TMVCP template-based nanocomposites allowing for a broader application of them. (paper)

  6. Radiochemical identification of the kil gene product of bacteriophage lambda

    International Nuclear Information System (INIS)

    Greer, H.; Ausubel, F.M.

    1979-01-01

    The coliphage lambda kil gene product has been identified using a differential labeling technique . The kil gene polypeptide has a molecular weight of about 16,000, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Gel filtration of the kil protein indicates that it may exist as a tetramer in native form

  7. On the radiochemical purity of elementary 35S with high specific activity

    International Nuclear Information System (INIS)

    Todorovsky, D.S.; Kostadinov, K.N.; Efremova, Yu.N.

    1979-01-01

    Radiochemical composition and chemical changes with increasing storage time of benzene solutions and of solid species of elementary 35 S with high specific activity are studied. The dependence of the stability on the specific activity and the radioactive concentration is shown and some tentative limits are given for permissible storage periods. (author)

  8. Determination of the radiochemical purity of phosphorus-32 and tritium-labeled diisopropylphosphorofluoridate (DFP)

    International Nuclear Information System (INIS)

    Christopher, R.E.; Sheppard, G.

    1975-01-01

    A method is described for the determination of the radiochemical purity of labeled diisopropylphosphorofluoridate (DFP), based on the irreversible inhibition reaction with the enzyme α-chymotrypsin. The nature of the impurities in commercially available 32 P- and 3 H-labeled DFP is discussed

  9. Radiochemical assay for determination of dihydropyrimidinase activity using reversed-phase high-performance liquid chromatography

    NARCIS (Netherlands)

    van Kuilenburg, A. B.; van Lenthe, H.; van Gennip, A. H.

    1999-01-01

    A radiochemical assay was developed to measure the activity of dihydropyrimidinase (DHP) in human liver homogenates. The method is based on the separation of radiolabeled dihydrouracil from N-carbamyl-beta-alanine by HPLC with on-line detection of radioactivity combined with detection of 14CO2 by

  10. Rapid and sensitive enzymatic-radiochemical assay for the determination of triglycerides

    International Nuclear Information System (INIS)

    Khoo, J.C.; Miller, E.; Goldberg, D.I.

    1987-01-01

    An enzymatic-radiochemical method suitable for the determination of triglyceride levels of cells in culture is described. The method is based on the enzymatic hydrolysis of triglycerides to free fatty acids which then complex with 63 Ni. The method is rapid, accurate, and inexpensive. The procedure extends the sensitivity of triglyceride measurement to as low as 0.25 nanomoles

  11. Triphenyl phosphine oxide as a substoichiometric radiochemical reagent: Determination of thallium

    International Nuclear Information System (INIS)

    Reddy, P.C.; Polaiah, B.; Rangamannar, B.

    1991-01-01

    A rapid radiochemical method has been developed for the determination of microgram amounts of thallium based on the substoichiometric extraction of its ocmplex with triphenylphosphine oxide into benzene from 6 M sulphuric acid. 10-90 μg of thallium was determined with an average error of 2.06%. The effect of diverse metal ions on the extraction was studied. (orig.)

  12. Computer aided piping layout design in radiochemical plants- an improved software package

    International Nuclear Information System (INIS)

    Raju, R.P.; Siddiqui, H.R.

    1995-01-01

    A software package was developed and it was successfully implemented for the piping layout design of the four process cells of the Kalpakkam Reprocessing Project. This paper discusses in detail all the improvements and modifications that are being carried out in the package so that it becomes more meaningful and useful for implementation for the forthcoming radiochemical plants

  13. Evaluation of radiochemical purities of some radiopharmaceuticals in Shiraz Namazi teaching hospital

    Directory of Open Access Journals (Sweden)

    Hossein Sadeghpour

    2015-03-01

    Full Text Available Many radiopharmaceuticals, as a special group of drugs, are eventually prepared at the nuclear medicine departments of the hospitals. Therefore, their quality control procedures such as sterility tests, radionuclide, radiochemical and chemical purity should be carried out in the hospitals. In this study, radiochemical purity for more than 300 preparations of three different radiopharmaceutical formulations from commercial kits were tested using instant thin layer chromatography. The formulations 99mTc-DTPA, 99mTc-MDP and 99mTc-MIBI were obtained from Pars Isotope Co. Several paper chromatographic systems including standard and factory recommended thin layer chromatography systems were used in this study. In addition different equipments for detection of radioactivity in paper chromatography like gamma camera and dose calibrator were used. The results showed that the most observed impurities were hydrolyzed reduced technetium (HR-Tc. There were no significant differences between calculated 99mTc-MIBI radiochemical purities when the radioactive detection device was gamma camera instead of dose calibrator. In case of 99mTc-DTPA and 99mTc-MDP, there were significant differences in detection of HR-Tc. On the contrary, no significant differences in free pertechnetate were observed when package insert procedures for quality control were used instead of those recommended in the references. Finally, we observed that the package insert procedures for quality control can offer higher radiochemical purities.

  14. Radiochemical separation and alpha-spectrometric determination of Americium in different matrixes

    International Nuclear Information System (INIS)

    Radenkovic, M.; Joksic, J.; Paligoric, D.

    2009-01-01

    A method of separation and alpha-spectrometric determination of americium, developed in VINCA Institute of Nuclear Sciences is described in the paper. The procedure is convenient to be used for 241 Am determination in environmental matrixes as well as samples of human origin if a very small concentrations are expected, using 243 Am as a tracer for radiochemical yield recovery. (author) [sr

  15. The radiochemical purity of radiotracers as the criterion of their usefulness in investigations of hydrocarbons distillation

    International Nuclear Information System (INIS)

    Kolaczkowski, M.

    1976-01-01

    Among numerous analytic techniques of separation and analysis of radioactive products gas radiochromatography has proved to be particularly suitable. Organic bromides labelled with 82 Br-radionuclide are investigated. The analytic gas chromatographs equipped with appropriately constructed radiochromatographic attachments are used. The results of radiochemical purity determination of radiotracers are compared for various techniques. (author)

  16. Analytical and radiochemical methods in the stability study of the MIBI-Sm

    International Nuclear Information System (INIS)

    Isaac, M.; Leyva, R.; Gamboa, R.; Turino, D.

    1997-01-01

    99mT c-MIBI is a radiopharmaceutical imaging agent useful for assessing myocardial perfusion. This paper presents the stability results obtained in the radiochemical purity and others parameters in the quality control of 5 batch during more than 2 years

  17. Biliary lipid composition and gallstone formation in rabbits fed on soy protein, cholesterol, casein and modified casein.

    OpenAIRE

    Ozben, T

    1989-01-01

    In four experimental groups, rabbits were fed on diets containing soy beans, soy beans plus cholesterol (1%, w/w), casein and modified casein for 8 weeks. Biliary lipid levels, lithogenic-index values and the rate of gallstone formation were determined. The highest mean relative concentrations (mol%) of cholesterol and phospholipid were found in the soy bean + cholesterol group, and the highest mean relative bile acid concentration was in the soy bean group. The lowest mean relative cholester...

  18. Detection of NT-pro BNP using fluorescent protein modified by streptavidin as a label in immunochromatographic assay

    Directory of Open Access Journals (Sweden)

    Haixia Li

    2016-12-01

    Full Text Available A novel fluorescent immunochromatographic assay for the detection of NT-proBNP in human serum has been developed. Based on a sandwich-type immunoassay format, analytes in samples were captured by one monoclonal antibody labeled with fluorescent protein and “sandwiched” by another monoclonal antibody immobilized on the nitrocellulose membrane, the fluorescence and concentration of analytes were measured and then calculated by fluoroanalyzer. The fluorescent protein is a fusion protein and was prepared through the application of Streptavidin gene SA, β subunit cpcB of Phycocyanin, lyase alr0617, and phycoerythrobilin synthetase gene ho1, pebA, pebB for covalent binding. It is characterized with higher stability, good solubility in water and it is not easy to quench fluorescence. Take the advantages of fluorescent protein, the immunochromatographic assay exhibited a wide linear range for NT-proBNP from 200 pg ml−1 to 26,000 pg ml−1, with a detection limit of 47 pg ml−1 under optimal conditions. Compared with chemiluminescence immunoassay (CLIA, 131 human serum samples were analyzed and the correlation coefficient of the developed immunoassay was 0.978. These results demonstrated that fluorescent immunochromatographic assay is a more rapid, sensitive, specific method and could be developed into a platform for more biomarkers determination in clinical practice. Keywords: NT-pro BNP, Fluorescent protein, Immunochromatographic assay

  19. UVA Light-excited Kynurenines Oxidize Ascorbate and Modify Lens Proteins through the Formation of Advanced Glycation End Products

    Science.gov (United States)

    Linetsky, Mikhail; Raghavan, Cibin T.; Johar, Kaid; Fan, Xingjun; Monnier, Vincent M.; Vasavada, Abhay R.; Nagaraj, Ram H.

    2014-01-01

    Advanced glycation end products (AGEs) contribute to lens protein pigmentation and cross-linking during aging and cataract formation. In vitro experiments have shown that ascorbate (ASC) oxidation products can form AGEs in proteins. However, the mechanisms of ASC oxidation and AGE formation in the human lens are poorly understood. Kynurenines are tryptophan oxidation products produced from the indoleamine 2,3-dioxygenase (IDO)-mediated kynurenine pathway and are present in the human lens. This study investigated the ability of UVA light-excited kynurenines to photooxidize ASC and to form AGEs in lens proteins. UVA light-excited kynurenines in both free and protein-bound forms rapidly oxidized ASC, and such oxidation occurred even in the absence of oxygen. High levels of GSH inhibited but did not completely block ASC oxidation. Upon UVA irradiation, pigmented proteins from human cataractous lenses also oxidized ASC. When exposed to UVA light (320–400 nm, 100 milliwatts/cm2, 45 min to 2 h), young human lenses (20–36 years), which contain high levels of free kynurenines, lost a significant portion of their ASC content and accumulated AGEs. A similar formation of AGEs was observed in UVA-irradiated lenses from human IDO/human sodium-dependent vitamin C transporter-2 mice, which contain high levels of kynurenines and ASC. Our data suggest that kynurenine-mediated ASC oxidation followed by AGE formation may be an important mechanism for lens aging and the development of senile cataracts in humans. PMID:24798334

  20. The Northern Marshall Islands radiological survey: A quality control program for radiochemical and gamma spectroscopy analysis

    International Nuclear Information System (INIS)

    Kehl, S.R.; Mount, M.E.; Robison, W.L.

    1995-09-01

    From 1979 to 1989, approximately 25,000 Post Northern Marshall Islands Radiological Survey (PNMIRS) samples were collected, and over 71,400 radiochemical and gamma spectroscopy analyses were performed to establish the concentration of 90 Sr, 137 Cs, 241 Am, and plutonium isotopes in soil, vegetation, fish, and animals in the Northern Marshall Islands. While the Low Level Gamma Counting Facility (B379) in the Health and Ecological Assessment (HEA) division accounted for over 80% of all gamma spectroscopy analyses, approximately 4889 radiochemical and 5437 gamma spectroscopy analyses were performed on 4784 samples of soil, vegetation, terrestrial animal, and marine organisms by outside laboratories. Four laboratories were used by Lawrence Livermore National Laboratory (LLNL) to perform the radiochemical analyses: Thermo Analytical Norcal, Richmond, California (TMA); Nuclear Energy Services, North Carolina State University (NCSU); Laboratory of Radiation Ecology, University of Washington (LRE); and Health and Ecological Assessment (HEA) division, LLNL, Livermore, California. Additionally, LRE and NCSU were used to perform gamma spectroscopy analyses. The analytical precision and accuracy were monitored by including blind duplicates and natural matrix standards in each group of samples analyzed. On the basis of reported analytical values for duplicates and standards, 88% of the gamma and 87% of the radiochemical analyses in this survey were accepted. By laboratory, 93% of the radiochemical analyses by TMA; 88% of the gamma-ray spectrometry and 100% of the radiochemistry analyses by NCSU; 89% of the gamma spectroscopy and 87% of the radiochemistry analyses by LRE; and 90% of the radiochemistry analyses performed by HEA's radiochemistry department were accepted

  1. A Legionella pneumophila effector protein encoded in a region of genomic plasticity binds to Dot/Icm-modified vacuoles.

    Directory of Open Access Journals (Sweden)

    Shira Ninio

    2009-01-01

    Full Text Available Legionella pneumophila is an opportunistic pathogen that can cause a severe pneumonia called Legionnaires' disease. In the environment, L. pneumophila is found in fresh water reservoirs in a large spectrum of environmental conditions, where the bacteria are able to replicate within a variety of protozoan hosts. To survive within eukaryotic cells, L. pneumophila require a type IV secretion system, designated Dot/Icm, that delivers bacterial effector proteins into the host cell cytoplasm. In recent years, a number of Dot/Icm substrate proteins have been identified; however, the function of most of these proteins remains unknown, and it is unclear why the bacterium maintains such a large repertoire of effectors to promote its survival. Here we investigate a region of the L. pneumophila chromosome that displays a high degree of plasticity among four sequenced L. pneumophila strains. Analysis of GC content suggests that several genes encoded in this region were acquired through horizontal gene transfer. Protein translocation studies establish that this region of genomic plasticity encodes for multiple Dot/Icm effectors. Ectopic expression studies in mammalian cells indicate that one of these substrates, a protein called PieA, has unique effector activities. PieA is an effector that can alter lysosome morphology and associates specifically with vacuoles that support L. pneumophila replication. It was determined that the association of PieA with vacuoles containing L. pneumophila requires modifications to the vacuole mediated by other Dot/Icm effectors. Thus, the localization properties of PieA reveal that the Dot/Icm system has the ability to spatially and temporally control the association of an effector with vacuoles containing L. pneumophila through activities mediated by other effector proteins.

  2. SAAFEC: Predicting the Effect of Single Point Mutations on Protein Folding Free Energy Using a Knowledge-Modified MM/PBSA Approach.

    Science.gov (United States)

    Getov, Ivan; Petukh, Marharyta; Alexov, Emil

    2016-04-07

    Folding free energy is an important biophysical characteristic of proteins that reflects the overall stability of the 3D structure of macromolecules. Changes in the amino acid sequence, naturally occurring or made in vitro, may affect the stability of the corresponding protein and thus could be associated with disease. Several approaches that predict the changes of the folding free energy caused by mutations have been proposed, but there is no method that is clearly superior to the others. The optimal goal is not only to accurately predict the folding free energy changes, but also to characterize the structural changes induced by mutations and the physical nature of the predicted folding free energy changes. Here we report a new method to predict the Single Amino Acid Folding free Energy Changes (SAAFEC) based on a knowledge-modified Molecular Mechanics Poisson-Boltzmann (MM/PBSA) approach. The method is comprised of two main components: a MM/PBSA component and a set of knowledge based terms delivered from a statistical study of the biophysical characteristics of proteins. The predictor utilizes a multiple linear regression model with weighted coefficients of various terms optimized against a set of experimental data. The aforementioned approach yields a correlation coefficient of 0.65 when benchmarked against 983 cases from 42 proteins in the ProTherm database. the webserver can be accessed via http://compbio.clemson.edu/SAAFEC/.

  3. Photochemical properties and sensor applications of modified yellow fluorescent protein (YFP) covalently attached to the surfaces of etched optical fibers (EOFs).

    Science.gov (United States)

    Veselov, Alexey A; Abraham, Bobin George; Lemmetyinen, Helge; Karp, Matti T; Tkachenko, Nikolai V

    2012-01-01

    Fluorescent proteins have the inherent ability to act as sensing components which function both in vitro and inside living cells. We describe here a novel study on a covalent site-specific bonding of fluorescent proteins to form self-assembled monolayers (SAMs) on the surface of etched optical fibers (EOFs). Deposition of fluorescent proteins on EOFs gives the opportunity to increase the interaction of guided light with deposited molecules relative to plane glass surfaces. The EOF modification is carried out by surface activation using 3-aminopropylthrimethoxysilane (APTMS) and bifunctional crosslinker sulfosuccinimidyl 4-[N-maleimidomethyl]cyclohexane-1-carboxylate (sulfo-SMCC) which exposes sulfhydryl-reactive maleimide groups followed by covalent site-specific coupling of modified yellow fluorescent protein (YFP). Steady-state and fluorescence lifetime measurements confirm the formation of SAM. The sensor applications of YPF SAMs on EOF are demonstrated by the gradual increase of emission intensity upon addition of Ca(2+) ions in the concentration range from a few tens of micromolars up to a few tens of millimolars. The studies on the effect of pH, divalent cations, denaturing agents, and proteases reveal the stability of YFP on EOFs at normal physiological conditions. However, treatments with 0.5% SDS at pH 8.5 and protease trypsin are found to denaturate or cleave the YFP from fiber surfaces.

  4. Devising of the method for the determination of small and very small amounts of cadmium in biological materials by radiochemical version of neutron activation analysis

    International Nuclear Information System (INIS)

    Dybczynski, R.; Samczynski, Z.

    1996-01-01

    The newly modified version of the method for the determination of cadmium in biological materials by radiochemical NAA based on selective post irradiation separation of Cd using ion exchange rasin Retardion 11A8 is presented. The conditions necessary for the selective retaining of Cd on the column exploiting both anionic and cationic ion exchange function of the resin have been discussed. Depending on the composition of the external solution, cadmium existing in the form of either anionic chloride complexes or cationic amine species is taken up by quaternary ammonium or carboxylate functional groups, respectively while accompanying elements are eluted. The elaborated method was further verified by determine Cd content in several certified biological reference materials using neutron activation analysis. The ion exchange separation procedure assures very high radiochemical purity of the cadmium fraction. Decontamination factors obtained for Mo, Sb, Na, Zn, Co, Sc amounted to 10 3 - 10 6 . Detection limit for Cd was 0.5 μg kg -1 . Analytical results show good agreement with the certified values. (author). 10 refs, 7 figs, 1 tab

  5. Immunotoxicological Evaluation of Genetically Modified Rice Expressing Cry1Ab/Ac Protein (TT51-1) by a 6-Month Feeding Study on Cynomolgus Monkeys

    Science.gov (United States)

    Tan, Xiaoyan; Zhou, Xiaobing; Tang, Yao; Lv, Jianjun; Zhang, Lin; Sun, Li; Yang, Yanwei; Miao, Yufa; Jiang, Hua; Chen, Gaofeng; Huang, Zhiying; Wang, Xue

    2016-01-01

    The present study was performed to evaluate the food safety of TT51-1, a new type of genetically modified rice that expresses the Cry1Ab/Ac protein (Bt toxin) and is highly resistant to most lepidopteran pests. Sixteen male and 16 female cynomolgus monkeys were randomly divided into four groups: conventional rice (non-genetically modified rice, non-GM rice), positive control, 17.5% genetically modified rice (GM rice) and 70% GM rice. Monkeys in the non-GM rice, positive control, and GM rice groups were fed on diets containing 70% non-GM rice, 17.5% GM rice or 70% GM rice, respectively, for 182 days, whereas animals in the positive group were intravenously injected with cyclophosphamide every other day for a total of four injections before the last treatment. Six months of treatment did not yield abnormal observations. Specifically, the following parameters did not significantly differ between the non-GM rice group and GM rice groups: body weight, food consumption, electrocardiogram, hematology, immuno-phenotyping of lymphocytes in the peripheral blood, mitogen-induced peripheral blood lymphocyte proliferation, splenocyte proliferation, KLH-T cell-dependent antibody response, organ weights and ratios, and histological appearance (p>0.05). Animals from the GM rice group differed from animals in the non-GM rice group (pGM rice. In conclusion, a 6-month feeding study of TT51-1 did not show adverse immunotoxicological effects on cynomolgus monkeys. PMID:27684490

  6. Structural study of the AOT reverse micellar system. Influence of attractive interactions induced by the solubilisation of native and modified proteins

    International Nuclear Information System (INIS)

    Cassin, Guillaume

    1994-01-01

    This research thesis reports the study of the influence of intra-micellar attractions on the thermodynamic behaviour of reverse micellar systems, as well as of the effects induced by the solubilisation of natives or modified proteins. The author proposes a model to explain the decrease of attractions between droplets when the volume fraction occupied by reverse micelles increases. This model which highlights the importance of depletion forces between reverse micelles, allows the building up of a theoretical relationship between the bonding parameter and the volume fraction of reverse micelles. In order to understand the appearance of an attractive term related to the solubilisation of native cytochrome-c in these systems, this protein has been chemically modified. The author highlights the role of the charge born by a micellar probe on the thermodynamic behaviour of micro-emulsions. Then, the author applies the model of dimerizing adhesive spheres to reverse micellar systems containing native cytochrome-c. He shows that theoretical predictions of this model are in agreement with obtained experimental results [fr

  7. Advanced glycation end products-modified proteins and oxidized LDL mediate down-regulation of leptin in mouse adipocytes via CD36

    International Nuclear Information System (INIS)

    Unno, Yuka; Sakai, Masakazu; Sakamoto, Yu-ichiro; Kuniyasu, Akihiko; Nakayama, Hitoshi; Nagai, Ryoji; Horiuchi, Seikoh

    2004-01-01

    Advanced glycation end products (AGE)-modified proteins as well as oxidized-LDL (Ox-LDL) undergo receptor-mediated endocytosis by CHO cells overexpressing CD36, a member of class B scavenger receptor family. The purpose of the present study was to examine the effects of glycolaldehyde-modified BSA (GA-BSA) as an AGE-ligand and Ox-LDL on leptin expression in adipocytes. GA-BSA decreased leptin expression at both protein and mRNA levels in 3T3-L1 adipocytes and mouse epididymal adipocytes. Ox-LDL showed a similar inhibitory effect on leptin expression in 3T3-L1 adipocytes, which effect was protected by N-acetylcysteine, a reactive oxygen species (ROS) inhibitor. Binding of 125 I-GA-BSA or 125 I-Ox-LDL to 3T3-L1 adipocytes and subsequent endocytic degradation were inhibited by a neutralizing anti-CD36 antibody. Furthermore, this antibody also suppressed Ox-LDL-induced leptin down-regulation. These results clarify that the interaction of GA-BSA and Ox-LDL with CD36 leads to down-regulation of leptin expression via ROS system(s) in 3T3-L1 adipocytes, suggesting that a potential link of AGE- and/or Ox-LDL-induced leptin down-regulation might be linked to insulin-sensitivity in metabolic syndrome

  8. Colon luminal content and epithelial cell morphology are markedly modified in rats fed with a high-protein diet.

    Science.gov (United States)

    Andriamihaja, Mireille; Davila, Anne-Marie; Eklou-Lawson, Mamy; Petit, Nathalie; Delpal, Serge; Allek, Fadhila; Blais, Anne; Delteil, Corine; Tomé, Daniel; Blachier, François

    2010-11-01

    Hyperproteic diets are used in human nutrition to obtain body weight reduction. Although increased protein ingestion results in an increased transfer of proteins from the small to the large intestine, there is little information on the consequences of the use of such diets on the composition of large intestine content and on epithelial cell morphology and metabolism. Rats were fed for 15 days with either a normoproteic (NP, 14% protein) or a hyperproteic isocaloric diet (HP, 53% protein), and absorptive colonocytes were observed by electron microscopy or isolated for enzyme activity studies. The colonic luminal content was recovered for biochemical analysis. Absorbing colonocytes were characterized by a 1.7-fold reduction in the height of the brush-border membranes (P = 0.0001) after HP diet consumption when compared with NP. This coincided in the whole colon content of HP animals with a 1.8-fold higher mass content (P = 0.0020), a 2.2-fold higher water content (P = 0.0240), a 5.2-fold higher protease activity (P = 0.0104), a 5.5-fold higher ammonia content (P = 0.0008), and a more than twofold higher propionate, valerate, isobutyrate, and isovalerate content (P hyperproteic diet ingestion causes marked changes both in the luminal environment of colonocytes and in the characteristics of these cells, demonstrating that hyperproteic diet interferes with colonocyte metabolism and morphology. Possible causal relationships between energy metabolism, reduced height of colonocyte brush-border membranes, and reduced water absorption are discussed.

  9. Detection of genetically modified organisms in foods by protein- and DNA-based techniques : bridging the methods

    NARCIS (Netherlands)

    Duijn, G. van; Biert, R. van; Bleeker-Marcelis, H.; Boeijen, I. van; Adan, A.J.; Jhakrie, S.; Hessing, M.

    2002-01-01

    According to European Commission (EC) Regulation 1139/98, foods and food ingredients that are to be delivered to the final consumer in which either protein or DNA resulting from genetic modification is present, shall be subject to additional specific labeling requirements. Since 1994, genetically

  10. Effect of addition of thermally modified cowpea protein on sensory acceptability and textural properties of wheat bread and sponge cake.

    Science.gov (United States)

    Campbell, Lydia; Euston, Stephen R; Ahmed, Mohamed A

    2016-03-01

    This paper investigates the sensory acceptability and textural properties of leavened wheat bread and sponge cake fortified with cow protein isolates that had been denatured and glycated by thermal treatment. Defatted cowpea flour was prepared from cow pea beans and the protein isolate was prepared (CPI) and thermally denatured (DCPI). To prepare glycated cowpea protein isolate (GCPI) the cowpea flour slurry was heat treated before isolation of the protein. CPI was more susceptible to thermal denaturation than GCPI as determined by turbidity and sulphydryl groups resulting in greater loss of solubility. This is attributed to the higher glycation degree and higher carbohydrate content of GCPI as demonstrated by glycoprotein staining of SDS PAGE gels. Water absorption of bread dough was significantly enhanced by DCPI and to a larger extent GCPI compared to the control, resulting in softer texture. CPI resulted in significantly increased crumb hardness in baked bread than the control whereas DCPI or GCPI resulted in significantly softer crumb. Bread fortified with 4% DCPI or GCPI was similar to control as regards sensory and textural properties whereas 4% CPI was significantly different, limiting its inclusion level to 2%. There was a trend for higher sensory acceptability scores for GCPI containing bread compared DCPI. Whole egg was replaced by 20% by GCPI (3.5%) in sponge cake without affecting the sensory acceptability, whereas CPI and DCPI supplemented cakes were significantly different than the control. Copyright © 2015 Elsevier Ltd. All rights reserved.

  11. Magnetic graphene oxide modified with choline chloride-based deep eutectic solvent for the solid-phase extraction of protein

    Energy Technology Data Exchange (ETDEWEB)

    Huang, Yanhua; Wang, Yuzhi, E-mail: wyzss@hnu.edu.cn; Pan, Qi; Wang, Ying; Ding, Xueqin; Xu, Kaijia; Li, Na; Wen, Qian

    2015-06-02

    Highlights: • A strategy for extraction of protein based on DES-coated magnetic graphene oxide. • The deep eutectic solvents were based on choline chloride. • Bovine serum albumin was used as the analyte. • The material prepared works for the acidic but not the basic or the neutral proteins. - Abstract: Four kinds of green deep eutectic solvents (DESs) based on choline chloride (ChCl) have been synthesized and coated on the surface of magnetic graphene oxide (Fe{sub 3}O{sub 4}@GO) to form Fe{sub 3}O{sub 4}@GO-DES for the magnetic solid-phase extraction of protein. X-ray diffraction (XRD), vibrating sample magnetometer (VSM), Fourier transform infrared spectrometry (FTIR), field emission scanning electron microscopy (FESEM) and thermal gravimetric analysis (TGA) were employed to characterize Fe{sub 3}O{sub 4}@GO-DES, and the results indicated the successful preparation of Fe{sub 3}O{sub 4}@GO-DES. The UV–vis spectrophotometer was used to measure the concentration of protein after extraction. Single factor experiments proved that the extraction amount was influenced by the types of DESs, solution temperature, solution ionic strength, extraction time, protein concentration and the amount of Fe{sub 3}O{sub 4}@GO-DES. Comparison of Fe{sub 3}O{sub 4}@GO and Fe{sub 3}O{sub 4}@GO-DES was carried out by extracting bovine serum albumin, ovalbumin, bovine hemoglobin and lysozyme. The experimental results showed that the proposed Fe{sub 3}O{sub 4}@GO-DES performs better than Fe{sub 3}O{sub 4}@GO in the extraction of acidic protein. Desorption of protein was carried out by eluting the solid extractant with 0.005 mol L{sup −1} Na{sub 2}HPO{sub 4} contained 1 mol L{sup −1} NaCl. The obtained elution efficiency was about 90.9%. Attributed to the convenient magnetic separation, the solid extractant could be easily recycled.

  12. Magnetic graphene oxide modified with choline chloride-based deep eutectic solvent for the solid-phase extraction of protein.

    Science.gov (United States)

    Huang, Yanhua; Wang, Yuzhi; Pan, Qi; Wang, Ying; Ding, Xueqin; Xu, Kaijia; Li, Na; Wen, Qian

    2015-06-02

    Four kinds of green deep eutectic solvents (DESs) based on choline chloride (ChCl) have been synthesized and coated on the surface of magnetic graphene oxide (Fe3O4@GO) to form Fe3O4@GO-DES for the magnetic solid-phase extraction of protein. X-ray diffraction (XRD), vibrating sample magnetometer (VSM), Fourier transform infrared spectrometry (FTIR), field emission scanning electron microscopy (FESEM) and thermal gravimetric analysis (TGA) were employed to characterize Fe3O4@GO-DES, and the results indicated the successful preparation of Fe3O4@GO-DES. The UV-vis spectrophotometer was used to measure the concentration of protein after extraction. Single factor experiments proved that the extraction amount was influenced by the types of DESs, solution temperature, solution ionic strength, extraction time, protein concentration and the amount of Fe3O4@GO-DES. Comparison of Fe3O4@GO and Fe3O4@GO-DES was carried out by extracting bovine serum albumin, ovalbumin, bovine hemoglobin and lysozyme. The experimental results showed that the proposed Fe3O4@GO-DES performs better than Fe3O4@GO in the extraction of acidic protein. Desorption of protein was carried out by eluting the solid extractant with 0.005 mol L(-1) Na2HPO4 contained 1 mol L(-1) NaCl. The obtained elution efficiency was about 90.9%. Attributed to the convenient magnetic separation, the solid extractant could be easily recycled. Copyright © 2015 Elsevier B.V. All rights reserved.

  13. Identification of Atg2 and ArfGAP1 as Candidate Genetic Modifiers of the Eye Pigmentation Phenotype of Adaptor Protein-3 (AP-3 Mutants in Drosophila melanogaster.

    Directory of Open Access Journals (Sweden)

    Imilce A Rodriguez-Fernandez

    Full Text Available The Adaptor Protein (AP-3 complex is an evolutionary conserved, molecular sorting device that mediates the intracellular trafficking of proteins to lysosomes and related organelles. Genetic defects in AP-3 subunits lead to impaired biogenesis of lysosome-related organelles (LROs such as mammalian melanosomes and insect eye pigment granules. In this work, we have performed a forward screening for genetic modifiers of AP-3 function in the fruit fly, Drosophila melanogaster. Specifically, we have tested collections of large multi-gene deletions--which together covered most of the autosomal chromosomes-to identify chromosomal regions that, when deleted in single copy, enhanced or ameliorated the eye pigmentation phenotype of two independent AP-3 subunit mutants. Fine-mapping led us to define two non-overlapping, relatively small critical regions within fly chromosome 3. The first critical region included the Atg2 gene, which encodes a conserved protein involved in autophagy. Loss of one functional copy of Atg2 ameliorated the pigmentation defects of mutants in AP-3 subunits as well as in two other genes previously implicated in LRO biogenesis, namely Blos1 and lightoid, and even increased the eye pigment content of wild-type flies. The second critical region included the ArfGAP1 gene, which encodes a conserved GTPase-activating protein with specificity towards GTPases of the Arf family. Loss of a single functional copy of the ArfGAP1 gene ameliorated the pigmentation phenotype of AP-3 mutants but did not to modify the eye pigmentation of wild-type flies or mutants in Blos1 or lightoid. Strikingly, loss of the second functional copy of the gene did not modify the phenotype of AP-3 mutants any further but elicited early lethality in males and abnormal eye morphology when combined with mutations in Blos1 and lightoid, respectively. These results provide genetic evidence for new functional links connecting the machinery for biogenesis of LROs with

  14. Effect of type and content of modified montmorillonite on the structure and properties of bio-nanocomposite films based on soy protein isolate and montmorillonite.

    Science.gov (United States)

    Kumar, P; Sandeep, K P; Alavi, S; Truong, V D; Gorga, R E

    2010-06-01

    The nonbiodegradable and nonrenewable nature of plastic packaging has led to a renewed interest in packaging materials based on bio-nanocomposites (biopolymer matrix reinforced with nanoparticles such as layered silicates). Bio-nanocomposite films based on soy protein isolate (SPI) and modified montmorillonite (MMT) were prepared using melt extrusion. The effect of different type (Cloisite 20A and Cloisite 30B) and content (0% to 15%) of modified MMT on the structure (degree of intercalation and exfoliation) and properties (color, mechanical, dynamic mechanical, thermal stability, and water vapor permeability) of SPI-MMT bio-nanocomposite films were investigated. Extrusion of SPI and modified MMTs resulted in bio-nanocomposites with exfoliated structures at lower MMT content (5%). At higher MMT content (15%), the structure of bio-nanocomposites ranged from intercalated for Cloisite 20A to disordered intercalated for Cloisite 30B. At an MMT content of 5%, bio-nanocomposite films based on modified MMTs (Cloisite 20A and Cloisite 30B) had better mechanical (tensile strength and percent elongation at break), dynamic mechanical (glass transition temperature and storage modulus), and water barrier properties as compared to those based on natural MMT (Cloisite Na(+)). Bio-nanocomposite films based on 10% Cloisite 30B had mechanical properties comparable to those of some of the plastics that are currently used in food packaging applications. However, much higher WVP values of these films as compared to those of existing plastics might limit the application of these films to packaging of high moisture foods such as fresh fruits and vegetables.

  15. Relationship between serum high-sensitivity C-reactive protein and modified TOAST classification as well as OCSP subtypes in patients with acute ischemic stroke

    Directory of Open Access Journals (Sweden)

    Hua-jun CHANG

    2014-10-01

    Full Text Available This paper aims to investigate the relationship between serum high-sensitivity C-reactive protein (hs-CRP level and modified TOAST classification as well as OCSP subtypes in patients with acute ischemic stroke. Serum hs-CRP was measured in 240 patients with acute ischemic stroke and 120 normal controls. All patients were classified according to modified TOAST classification and OCSP criteria. Serum hs-CRP levels in acute ischemic stroke group were significantly higher than those in normal control group [(13.68 ± 6.92 mg/L vs (3.98 ± 0.76 mg/L; t = 6.922, P = 0.002]. Among modified TOAST subtypes, the highest serum hs-CRP level was in cardioembolism (CE group [(16.82 ± 6.16 mg/L], followed by arterothrombosis (AT group [(15.17 ± 5.68 mg/L], stroke of undetermined etiology (SUD group [(10.06 ± 3.89 mg/L] and small artery disease (SAD group [(9.86 ± 3.75 mg/L, P = 0.027]. Among OCSP subtypes, the highest serum hs-CRP level was in total anterior circulation infarct (TACI group [(17.02 ± 6.98 mg/L], followed by posterior circulation infarct (POCI group [(15.91 ± 7.12 mg/L], partial anterior circulation infarct (PACI group [(12.83 ± 4.95 mg/L] and lacunar infarct (LACI group [(10.61 ± 5.73 mg/L, P = 0.005]. Serum hs-CRP levels are various in different modified TOAST and OCSP subtypes, which may reflect etiological and pathophysiological diversity of acute ischemic stroke, guide clinical treatment and help to predict prognosis. doi: 10.3969/j.issn.1672-6731.2014.10.013

  16. The analysis of thallium in geological materials by radiochemical neutron activation and x-ray fluorescence spectrometry: a comparison

    Energy Technology Data Exchange (ETDEWEB)

    McGoldrick, P J; Robinson, P [Tasmania Univ., Sandy Bay, TAS (Australia)

    1994-12-31

    Carrier-based radiochemical neutron activation (RNAA) is a precise and accurate technique for the analysis of Tl in geological materials. For about a decade, until the mid-80s, a procedure modified from Keays et al. (1974) was used at the University of Melbourne to analyse for Tl in a wide variety of geological materials. Samples of powdered rock weighing several hundred milligrams each were irradiated in HIFAR for between 12 hours and 1 week, and subsequently fused with a sodium hydroxide - sodium peroxide mixture and several milligrams of inactive Tl carrier. Following acid digestion of the fusion mixture anion exchange resin was used to separate Tl from the major radioactive rock constituents. The Tl was then stripped from the resin and purified as thallium iodide and a yield measured gravimetrically. Activity from {sup 204}Tl (a {beta}-emitter with a 3 8 year half-life) was measured and Tl determined by reference to pure chemical standards irradiated and processed along with the unkowns. Detection limits for the longer irradiations were about one part per billion. Precision was monitored by repeat analyses of `internal standard` rocks and was estimated to be about five to ten percent (one standard deviation). On the other hand, X-ray fluorescence spectrometry (XRF) was seen as an excellent cost-effective alternative for thallium analysis in geological samples, down to 1 ppm. 6 refs. 1 tab., 1 fig.

  17. The analysis of thallium in geological materials by radiochemical neutron activation and x-ray fluorescence spectrometry: a comparison

    Energy Technology Data Exchange (ETDEWEB)

    McGoldrick, P.J.; Robinson, P. [Tasmania Univ., Sandy Bay, TAS (Australia)

    1993-12-31

    Carrier-based radiochemical neutron activation (RNAA) is a precise and accurate technique for the analysis of Tl in geological materials. For about a decade, until the mid-80s, a procedure modified from Keays et al. (1974) was used at the University of Melbourne to analyse for Tl in a wide variety of geological materials. Samples of powdered rock weighing several hundred milligrams each were irradiated in HIFAR for between 12 hours and 1 week, and subsequently fused with a sodium hydroxide - sodium peroxide mixture and several milligrams of inactive Tl carrier. Following acid digestion of the fusion mixture anion exchange resin was used to separate Tl from the major radioactive rock constituents. The Tl was then stripped from the resin and purified as thallium iodide and a yield measured gravimetrically. Activity from {sup 204}Tl (a {beta}-emitter with a 3 8 year half-life) was measured and Tl determined by reference to pure chemical standards irradiated and processed along with the unkowns. Detection limits for the longer irradiations were about one part per billion. Precision was monitored by repeat analyses of `internal standard` rocks and was estimated to be about five to ten percent (one standard deviation). On the other hand, X-ray fluorescence spectrometry (XRF) was seen as an excellent cost-effective alternative for thallium analysis in geological samples, down to 1 ppm. 6 refs. 1 tab., 1 fig.

  18. A 52-week safety study in cynomolgus macaques for genetically modified rice expressing Cry1Ab/1Ac protein.

    Science.gov (United States)

    Mao, Jie; Sun, Xing; Cheng, Jian-Hua; Shi, Yong-Jie; Wang, Xin-Zheng; Qin, Jun-Jie; Sang, Zhi-Hong; He, Kun; Xia, Qing

    2016-09-01

    A 52-week feeding study in cynomolgus macaques was carried out to evaluate the safety of Bt rice Huahui 1 (HH1), a transgenic rice line expressing Cry1Ab/1Ac protein. Monkeys were fed a diet with 20% or 60% HH1 rice, 20% or 60% parental rice (Minghui 63, MH63), normal diet, normal diet spiked with purified recombinant Cry1Ab/1Ac fusion protein or bovine serum albumin (BSA) respectively. During the feeding trail, clinical observations were conducted daily, and multiple parameters, including body weight, body temperature, electrocardiogram, hematology, blood biochemistry, serum metabolome and gut microbiome were examined at regular intervals. Upon sacrifice, the organs were weighted, and the macroscopic, microscopic and electron microscopic examinations were performed. The results show no adverse or toxic effects of Bt rice HH1 or Cry1Ab/1Ac fusion protein on monkeys. Therefore, the present 52-week primate feeding study suggests that the transgenic rice containing Cry 1Ab/1Ac is equivalent to its parental rice line MH63. Copyright © 2016 Elsevier Ltd. All rights reserved.

  19. HPV16-E2 protein modifies self-renewal and differentiation rate in progenitor cells of human immortalized keratinocytes.

    Science.gov (United States)

    Domínguez-Catzín, Victoria; Reveles-Espinoza, Alicia-María; Sánchez-Ramos, Janet; Cruz-Cadena, Raúl; Lemus-Hernández, Diana; Garrido, Efraín

    2017-04-03

    Cervical cancer is the fourth cause of death worldwide by cancer in women and is a disease associated to persistent infection with human papillomavirus (HPV), particularly from two high-risk types HPV16 and 18. The virus initiates its replicative cycle infecting cells located in the basal layer of the epithelium, where a small population of epithelial stem cells is located performing important functions of renewal and maintenance of the tissue. Viral E2 gene is one of the first expressed after infection and plays relevant roles in the replicative cycle of the virus, modifying fundamental processes in the infected cells. Thus, the aim of the present study was to demonstrate the presence of hierarchic subpopulations in HaCaT cell line and evaluate the effect of HPV16-E2 expression, on their biological processes. HaCaT-HPV16-E2 cells were generated by transduction of HaCaT cell line with a lentiviral vector. The α6-integrin-CD71 expression profile was established by immunostaining and flow cytometric analysis. After sorting, cell subpopulations were analyzed in biological assays for self-renewal, clonogenicity and expression of stemness factors (RT-qPCR). We identified in HaCaT cell line three different subpopulations that correspond to early differentiated cells (α6-integrin dim ), transitory amplifying cells (α6-integrin bri /CD71 bri ) and progenitor cells (α6-integrin bri /CD71 dim ). The last subpopulation showed stem cell characteristics, such as self-renewal ability, clonogenicity and expression of the well-known stem cell factors SOX2, OCT4 and NANOG, suggesting they are stem-like cells. Interestingly, the expression of HPV16-E2 in HaCaT cells changed its α6-integrin-CD71 immunophenotype modifying the relative abundance of the cell subpopulations, reducing significantly the percentage of α6-integrin bri /CD71 dim cells. Moreover, the expression of the stem cell markers was also modified, increasing the expression of SOX2 and NANOG, but decreasing notably

  20. OPERATIONAL EXPERIENCE: UPGRADED MPC AND A SYSTEMS FOR THE RADIOCHEMICAL PLANT OF THE SIBERIAN CHEMICAL COMBINE

    International Nuclear Information System (INIS)

    RODRIGUEZ, C.; GOLOSKOKOV, I.; FISHBONE, L.; GOODEY, K.; LOOMIS, M.; CRAIN, B. JR.; LARSEN, R.

    2003-01-01

    The success of reducing the risk of nuclear proliferation through physical protection and material control/accounting systems depends upon the development of an effective design that includes consideration of the objectives of the systems and the resources available to implement the design. Included among the objectives of the design are facility characterization, definition of threat, and identification of targets. When considering resources, the designer must consider funds available, rapid low-cost elements, technology elements, human resources, and the availability of resources to sustain operation of the end system. The Siberian Chemical Combine (SCC) is a multi-function nuclear facility located in the Tomsk region of Siberia, Russia. Beginning in 1996, SCC joined with the United States Department of Energy (US/DOE) Material Protection, Control, and Accounting (MPC and A) Program to develop and implement MPC and A upgrades for the Radiochemical, Chemical Metallurgical, Conversion, Uranium Enrichment, and Reactor Plants of the SCC. At the Radiochemical Plant the MPC and A design and implementation process has been largely completed for the Plutonium Storage Facility and related areas of the Radiochemical Plant. Design and implementation of upgrades for the Radiochemical Plant include rapid physical protection upgrades such as bricking up of doors and windows, and installation of security-hardened doors. Rapid material control and accounting upgrades include installation of modern balances and bar code equipment. Comprehensive MPC and A upgrades include the installation of access controls to sensitive areas of the Plant, alarm communication and display (AC and D) systems to detect and annunciate alarm conditions, closed circuit (CCTV) systems to assess alarm conditions, central and secondary alarm station upgrades that enable security forces to assess and respond to alarm conditions, material control and accounting upgrades that include upgraded physical

  1. Expression characterization and functional implication of the collagen-modifying Leprecan proteins in mouse gonadal tissue and mature sperm

    Directory of Open Access Journals (Sweden)

    Sarah M. Zimmerman

    2018-02-01

    Full Text Available The Leprecan protein family which includes the prolyl 3-hydroxylase enzymes (P3H1, P3H2, and P3H3, the closely related cartilage-associated protein (CRTAP, and SC65 (Synaptonemal complex 65, aka P3H4, LEPREL4, is involved in the post-translational modification of fibrillar collagens. Mutations in CRTAP, P3H1 and P3H2 cause human genetic diseases. We recently showed that SC65 forms a stable complex in the endoplasmic reticulum with P3H3 and lysyl hydroxylase 1 and that loss of this complex leads to defective collagen lysyl hydroxylation and causes low bone mass and skin fragility. Interestingly, SC65 was initially described as a synaptonemal complex-associated protein, suggesting a potential additional role in germline cells. In the present study, we describe the expression of SC65, CRTAP and other Leprecan proteins in postnatal mouse reproductive organs. We detect SC65 expression in peritubular cells of testis up to 4 weeks of age but not in cells within seminiferous tubules, while its expression is maintained in ovarian follicles until adulthood. Similar to bone and skin, SC65 and P3H3 are also tightly co-expressed in testis and ovary. Moreover, we show that CRTAP, a protein normally involved in collagen prolyl 3-hydroxylation, is highly expressed in follicles and stroma of the ovary and in testes interstitial cells at 4 weeks of age, germline cells and mature sperm. Importantly, CrtapKO mice have a mild but significant increase in morphologically abnormal mature sperm (17% increase compared to WT. These data suggest a role for the Leprecans in the post-translational modification of collagens expressed in the stroma of the reproductive organs. While we could not confirm that SC65 is part of the synaptonemal complex, the expression of CRTAP in the seminiferous tubules and in mature sperm suggest a role in the testis germ cell lineage and sperm morphogenesis.

  2. Scientific and technical conference. Problems and horizons of development of chemical and radiochemical control in nuclear energetics. Collection of summaries of reports

    International Nuclear Information System (INIS)

    2001-01-01

    During scientific and technical conference on problems of development of chemical and radiochemical control in nuclear energetics following themes were considered: the problems of methodological and instrumental assurance of chemical and radiochemical control at working nuclear power plants and nuclear energetic units; modern conceptions of automation systems construction of chemical and radiochemical control on the basis of intellectual measuring channels; the ways of decision of generally system problems of organization and management of chemical and radiochemical control using computed technologies; the problems of certification of chemical and radiochemical methods of measuring in nuclear energetics [ru

  3. Drug-binding ability of human serum albumin at children with chronic virus hepatitis radiochemical definition method

    International Nuclear Information System (INIS)

    Kim, A.A.; Dadakhanov, J.A.; Djuraeva, G.T.; Shukurov, B.V.; Mavlyanov, I.R.

    2006-01-01

    Full text: The chronic virus hepatitis produces numerous abnormalities of liver function. The viruses of B, C, D, F and G hepatitis possess the ability to cause chronically proceeding diseases. Earlier we have found that binding ability of serum albumin at patients with acute forms of virus hepatitis is authentically reduced in comparison with the given parameters of control group. At an acute virus hepatitis B with middle severity the reducing of binding ability of serum albumin was observed at 70 % of patients. At an acute virus hepatitis A the reduce of binding ability of serum albumin is less expressed than at acute virus hepatitis B. At of chronic virus intoxication in human organism there is a formation and accumulation of toxic compounds in the excessive concentrations, which are not inherent to a normal metabolism. One of universal mechanisms of reaction of an organism on the increasing concentration of metabolism products is formation of complexes of various compounds with blood plasma proteins. The formation in an organism of endo- and exotoxins excessive concentrations results in blocking the binding centers of albumin molecule that causes the change of its complexing ability. The purpose of the present research: investigation of binding ability of serum albumin with use of radiochemical method at children with a chronic virus hepatitis B and C. Materials and methods. Under clinical observation there were 52 children in the age from 3 till 14 years. From them at 32 the chronic virus hepatitis B was confirmed, at 20 chronic virus - hepatitis C. Etiological diagnostics was carried out by definition of specific markers of a hepatitis B and C method IFA and PCR. Binding ability of serum albumin was defined by radiochemical method with use of the tritium labeled no-spa (drotaverine hydrochloride). The control group consists from 10 conditionally health children of similar age. Results and their discussion. The results of investigation have shown, that at a

  4. Development of Monoclonal Antibodies Recognizing Linear Epitope: Illustration by Three Bacillus thuringiensis Crystal Proteins of Genetically Modified Cotton, Maize, and Tobacco.

    Science.gov (United States)

    Cao, Zhen; Zhang, Wei; Ning, Xiangxue; Wang, Baomin; Liu, Yunjun; Li, Qing X

    2017-11-22

    Bacillus thuringiensis Cry1Ac, Cry1Ia1, and Cry1Ie are δ-endotoxin insecticidal proteins widely implemented in genetically modified organisms (GMO), such as cotton, maize, and potato. Western blot assay integrates electrophoresis separation power and antibody high specificity for monitoring specific exogenous proteins expressed in GMO. Procedures for evoking monoclonal antibody (mAb) for Western blot were poorly documented. In the present study, Cry1Ac partially denatured at 100 °C for 5 min was used as an immunogen to develop mAbs selectively recognizing a linear epitope of Cry1Ac for Western blot. mAb 5E9C6 and 3E6E2 selected with sandwich ELISA strongly recognized the heat semidenatured Cry1Ac. Particularly, 3E6E2 recognized both E. coli and cotton seed expressed Cry1Ac in Western blot. Such strategy of using partially denatured proteins as immunogens and using sandwich ELISA for mAb screening was also successfully demonstrated with production of mAbs against Cry1Ie for Western blot assay in maize.

  5. Continuous and intermittent transcranial magnetic theta burst stimulation modify tactile learning performance and cortical protein expression in the rat differently.

    Science.gov (United States)

    Mix, Annika; Benali, Alia; Eysel, Ulf T; Funke, Klaus

    2010-11-01

    Repetitive transcranial magnetic stimulation (rTMS) can modulate cortical excitability in a stimulus-frequency-dependent manner. Two kinds of theta burst stimulation (TBS) [intermittent TBS (iTBS) and continuous TBS (cTBS)] modulate human cortical excitability differently, with iTBS increasing it and cTBS decreasing it. In rats, we recently showed that this is accompanied by changes in the cortical expression of proteins related to the activity of inhibitory neurons. Expression levels of the calcium-binding protein parvalbumin (PV) and of the 67-kDa isoform of glutamic acid decarboxylase (GAD67) were strongly reduced following iTBS, but not cTBS, whereas both increased expression of the 65-kDa isoform of glutamic acid decarboxylase. In the present study, to investigate possible functional consequences, we applied iTBS and cTBS to rats learning a tactile discrimination task. Conscious rats received either verum or sham rTMS prior to the task. Finally, to investigate how rTMS and learning effects interact, protein expression was determined for cortical areas directly involved in the task and for those either not, or indirectly, involved. We found that iTBS, but not cTBS, improved learning and strongly reduced cortical PV and GAD67 expression. However, the combination of learning and iTBS prevented this effect in those cortical areas involved in the task, but not in unrelated areas. We conclude that the improved learning found following iTBS is a result of the interaction of two effects, possibly in a homeostatic manner: a general weakening of inhibition mediated by the fast-spiking interneurons, and re-established activity in those neurons specifically involved in the learning task, leading to enhanced contrast between learning-induced and background activity. © 2010 The Authors. European Journal of Neuroscience © 2010 Federation of European Neuroscience Societies and Blackwell Publishing Ltd.

  6. Approaches to assess IgE mediated allergy risks (sensitization and cross-reactivity) from new or modified dietary proteins

    DEFF Research Database (Denmark)

    Remington, B.; Broekman, H. C. H.; Blom, W. M.

    2018-01-01

    for new proteins, and especially to identify and characterise the risk of sensitization for IgE mediated allergy from oral exposure. Existing tools and tests are capable of assessing potential crossreactivity. However, there are few possibilities to assess the hazard due to de novo sensitization. The only...... methods available are in vivo models, but many limitations exist to use them for assessing risk. We conclude that there is a need to understand which criteria adequately define allergenicity for risk assessment purposes, and from these criteria develop a more suitable battery of tests to distinguish...

  7. Clinical utility of spot urine protein-to-creatinine ratio modified by estimated daily creatinine excretion in children.

    Science.gov (United States)

    Yang, Eun Mi; Yoon, Bo Ae; Kim, Soo Wan; Kim, Chan Jong

    2017-06-01

    The spot urine protein-to-creatinine ratio (UPCR) is widely used to predict 24-h urine protein (24-h UP) excretion. In patients with low daily urine creatinine excretion (UCr), however, the UPCR may overestimate 24-h UP. The aim of this study was to predict 24-h UP using UPCR adjusted by estimated 24-h UCr in children. This study included 442 children whose 24-h UP and spot UPCR were measured concomitantly. Estimated 24-h UCr was calculated using three previously existing equations. We estimated the 24-h UP excretion from UPCR by multiplying the estimated UCr. The results were compared with the measured 24-h UP. There was a strong correlation between UPCR and 24-h UP (r = 0.801, P < 0.001), and the correlation improved after multiplying the UPCR by the measured UCr (r = 0.847, P < 0.001). Using the estimated UCr rather than the measured UCr, there was high accuracy and strong correlation between the estimated UPCR weighted by the Cockcroft-Gault equation and 24-h UP. Improvement was also observed in the subgroup (proteinuria vs. non-proteinuria) analysis, particularly in the proteinuria group. The spot UPCR multiplied by the estimated UCr improved the accuracy of prediction of the 24-h UP in children.

  8. Trace determination of uranium and thorium in biological samples by radiochemical neutron activation analysis

    International Nuclear Information System (INIS)

    Benedik, Ljudmila; Repinc, Urska; Byrne, Anthony R.; Stegnar, Peter

    2002-01-01

    Radiochemical neutron activation analysis (RNAA) is an excellent method for determining uranium and thorium; it offers unique possibilities for their ultratrace analysis using selective radiochemical separations. Regarding the favourably sensitive nuclear characteristics of uranium and of thorium with respect to RNAA, but the different half-lives of their induced nuclides, two different approaches were used. In the first approach uranium and thorium were determined separately via 239 U, 239 Np and 233 Pa. In the second approach these elements were 239 239 233 determined simultaneously in a single sample using U and/or Np and Pa. Isolation of induced nuclides was based on separation by extraction and/or anion exchange chromatography. Chemical yields were measured in each sample aliquot using added 235 U, 238 Np and 231 Pa radioisotopic tracers. (author)

  9. Slifers revisited: a method for determining yields independent of radiochemical measurements

    International Nuclear Information System (INIS)

    Rambo, J.T.

    1976-01-01

    It would be very desirable if an independent method other than radiochemical measurement were available to determine the yields of low-yield events in the alluviums and tuffs of areas 2, 9, and 10 at the Nevada Test Site. The successful application of slifers to the measurement of yields from high-yield events suggests that under some conditions they may also be usable with low-yield events. This view is supported by the evidence discussed here, which is based on direct experience with slifer yield measurements for low-yield events in porous media. Suggested methods for improving slifer yield determinations and a method for determining yields independent of radiochemical measurements are offered

  10. Radiochemical methodologies applied to analytical characterization of low and intermediate level wastes from nuclear power plants

    Energy Technology Data Exchange (ETDEWEB)

    Monteiro, Roberto Pellacani G.; Júnior, Aluísio Souza R.; Kastner, Geraldo F.; Temba, Eliane S.C.; Oliveira, Thiago C. de; Amaral, Ângela M.; Franco, Milton B., E-mail: rpgm@cdtn.br, E-mail: reisas@cdtn.br, E-mail: gfk@cdtn.br, E-mail: esct@cdtn.br, E-mail: tco@cdtn.br, E-mail: ama@cdtn.br, E-mail: francom@cdtn.br [Centro de Desenvolvimento da Tecnologia Nuclear (CDTN/CNEN-MG), Belo Horizonte, MG (Brazil)

    2017-07-01

    The aim of this work is to present radiochemical methodologies developed at CDTN/CNEN in order to answer a program for isotopic inventory of radioactive wastes from Brazilian Nuclear Power Plants. In this program some radionuclides, {sup 3}H, {sup 14}C, {sup 55}Fe, {sup 59}Ni, {sup 63}Ni, {sup 90}Sr, {sup 93}Zr, {sup 94}Nb, {sup 99}Tc, {sup 129}I, {sup 235}U, {sup 238}U, {sup 238}Pu, {sup 239}+{sup 240}Pu, {sup 241}Pu, {sup 242}Pu, {sup 241}Am, {sup 242}Cm e {sup 243}+{sup 244}Cm, were determined in Low Level Wastes (LLW) and Intermediate Level Wastes (ILW) and a protocol of analytical methodologies based on radiochemical separation steps and spectrometric and nuclear techniques was established. (author)

  11. Requirement of radiochemical recovery determination for gross alpha and gross beta estimation in drinking water

    International Nuclear Information System (INIS)

    Raveendran, Nanda; Rao, D.D.; Hegde, A.G.

    2010-01-01

    Presence of radionuclides in drinking water which emits Alpha and Beta particles are the potential sources of internal exposure in drinking water. Gross alpha and gross beta determination in drinking water and packaged drinking water (PDW) as per BIS (Bureau of Indian standards) standards is discussed here. The methods have been tested to account for losses in the radiochemical procedures using radionuclides such as 137 Cs, 90 Sr, 226 Ra, 239 Pu, 243 Am, 232 U. The methods have also been validated in an IAEA proficiency test conducted during 2009. Monitoring of gross alpha and gross beta activity observed in drinking water/packaged drinking water from various states of India were within the limits set by BIS. Average radiochemical recoveries of 84% and 63% were obtained for gross α and gross β respectively. (author)

  12. Procedural and developmental aspects of a multielement automatic radiochemical machine, applied to neutron irradiated biomedical samples

    International Nuclear Information System (INIS)

    Iyengar, G.V.

    1976-06-01

    This report is intended to serve as a practical guide, elaborately describing the working details and some developmental work connected with an automatic multielement radiochemical machine based on thermal neutron activation analysis using ion exchange and partition chromatography. Some of the practical aspects and personal observations after much experience with this versatile multielement method, applied to investigate the elemental composition of different biomedical matrices, are summarized. Standard reference materials are analyzed, and the data are presented with a set of gamma-spectra obtained before and after chemical separation into convenient groups suitable for gamma spectroscopy. The samples analyzed included various human and animal tissues, body fluids, IAEA biological standard reference materials, and samples from the WHO/IAEA project on 'Trace elements in relation to cardiovascular diseases'. Simplified modifications of the radiochemical processing, suitable for fast and routine analysis of clinical samples have also been discussed. (orig.) [de

  13. Ultratrace determination of platinum in biological materials via neutron activation and radiochemical separation

    International Nuclear Information System (INIS)

    Zeisler, R.; Greenberg, R.R.

    1982-01-01

    A neutron activation analysis scheme based upon a radiochemical separation of the activation products has been developed. The method utilizes the inherent sensitivity of the activation reaction 198 Pt(n,ν) 199 Pt and counting of the daughter nuclide 199 Au. This nuclide is radiochemically separated from interfering activities by homogeneous precipitation as elemental gold. The remaining interference of the secondary reaction 197 Au(n,ν) 198 Au(n,ν) 199 Au from gold in the samples is quantitatively assessed and corrected. During this process accurate gold concentrations in the samples are obtained at ultratrace levels. The analysis scheme is applied to gold and platinum determinations in biological Standard Reference Materials and human liver specimens. Gold and platinum are determined at concentrations of 5x10 - 11 g/g, and at higher levels. (author)

  14. Radiochemical syntheses further radiopharmaceuticals for positron emission tomography and new strategies for their production

    CERN Document Server

    Kilbourn, Michael R; Kilbourn, Michael R

    2015-01-01

    This book describes methods and procedures for preparing PET radiopharmaceuticals, and highlights new methods for conducting radiochemical reactions with carbon-11 (C11) and fluorine-18 (F18), which are two of the most commonly used radionuclides in positron emission tomography (PET) imaging.     Provides reliable methods for radiochemical syntheses and reactions, including all essential information to duplicate the procedure     Eliminates the time-consuming process of searching journal articles and extracting pertinent details from lengthy experimental sections or supporting information     Focuses on an emerging and important area for pharmaceutical and medical applications     Encompasses technical, regulatory, and application aspects     Includes solid-phase radiochemistry, transition-metal catalyzed radiochemistry, microfluidics, click chemistry, green radiochemistry and new strategies for radiopharmaceutical quality control.

  15. Radiochemical neutron activation analysis for trace elements of basic ingredients of pan

    International Nuclear Information System (INIS)

    Zaidi, J.H.; Arif, M.; Fatima, I; Qureshi, I.H.

    2002-01-01

    Extensive use of pan, by one-tenth of world's population, entails the evaluation of trace element contents in its ingredients. Radiochemical neutron activation analysis (RNAA) was developed and successfully employed to determine the concentration of 36 trace elements (essential, toxic and nonessential) in its four basic ingredients, leaf of betel pepper, betel nut, catechu and lime. The radiochemical separation methodology has significantly improved the detection limits of most of these elements due to suppression of Compton background. Base-line values of certain toxic and essential elements in these ingredients is provided. The daily intake of essential and toxic elements through pan was estimated and compared with the recommended values. The cumulative intake of Mn is four times higher than the recommended value and that of toxic elements is well below the tolerance limits. (author)

  16. RADCHEM - Radiochemical procedures for the determination of Sr, U, Pu, Am and Cm

    Energy Technology Data Exchange (ETDEWEB)

    Sidhu, R. [Inst. for Energy Technology (Norway)

    2006-04-15

    An accurate determination of radionuclides from various sources in the environment is essential for assessment of the potential hazards and suitable countermeasures both in case of accidents, authorised release and routine surveillance. Reliable radiochemical separation and detection techniques are needed for accurate determination of alpha and beta emitters. Rapid analytical methods are needed in case of an accident for early decision-making. The objective of this project has been to compare and evaluate radiochemical procedures used at Nordic laboratories for the determination of strontium, uranium, plutonium, americium and curium. To gather detailed information on the procedures in use, a questionnaire regarding various aspects of radionuclide determination was developed and distributed to all (sixteen) relevant laboratories in the Nordic countries. The response and the procedures used by each laboratory were then discussed between those who answered the questionnaire. This report summaries the findings and gives recommendation on suitable practice. (au)

  17. Leakage evaluation in the PCV (Primary Containment Vessel) using chemical and radiochemical data

    International Nuclear Information System (INIS)

    Maeda, Katsuji; Nagasawa, Katsumi

    1998-01-01

    Keeping the reliability of nuclear power plant operation, the primary coolant leakage in the PCV is strictly restricted by the Technical Specifications. It is very important to detect an indication of leakage and estimate the source of leakage to provide countermeasures. Usually the indication of leakage will be detected by increase of drain flow in the PCV sump. There are some possibilities of leakage sources in the PCV, such as reactor water, main steam, condensate, feedwater and closed cooling water. The leakage source contain different chemical and radiochemical species. This means that the leakage source can be presumed and detected by using chemical information from the PCV atmosphere and sump water. To detect the leakage indication and the source quickly and exactly, the PCV Leakage Detection Expert System has been developed. This paper describes how to evaluate the leakage indication and source in the PCV by using chemical and radiochemical data. (author)

  18. RADCHEM - Radiochemical procedures for the determination of Sr, U, Pu, Am and Cm

    International Nuclear Information System (INIS)

    Sidhu, R.

    2006-04-01

    An accurate determination of radionuclides from various sources in the environment is essential for assessment of the potential hazards and suitable countermeasures both in case of accidents, authorised release and routine surveillance. Reliable radiochemical separation and detection techniques are needed for accurate determination of alpha and beta emitters. Rapid analytical methods are needed in case of an accident for early decision-making. The objective of this project has been to compare and evaluate radiochemical procedures used at Nordic laboratories for the determination of strontium, uranium, plutonium, americium and curium. To gather detailed information on the procedures in use, a questionnaire regarding various aspects of radionuclide determination was developed and distributed to all (sixteen) relevant laboratories in the Nordic countries. The response and the procedures used by each laboratory were then discussed between those who answered the questionnaire. This report summaries the findings and gives recommendation on suitable practice. (au)

  19. Rapid radiochemical methods for preparation of sup(99m)Tc labelled compounds

    International Nuclear Information System (INIS)

    Narasimhan, D.V.S.; Banodkar, S.M.; Kothari, K.; Mani, R.S.

    1981-01-01

    Several inorganic and organic compounds incorporating sup( 99 m)Tc are being extensively used for imaging various body organs. The preparation of these sup( 99 m)Tc compounds with the necessary purity requirements is carried out by controlled reduction of sup( 99 m)Tc-pertechnetate using Sn(II) ions as the reducing agent followed by complexation with various active ingredients. The authors here present procedures developed at Radiopharmaceuticals Section of BARC for preparing sup( 99 m)Tc-diphosphonate, sup( 99 m)Tc-glucoheptonate, sup( 99 m)Tc-albumin microspheres and sup( 99 m)Tc-phytate with high radiochemical purity. The paper also describes procedures for the preparation of freeze-dried kits for single step preparation of these compounds. The paper also describes the authors' experience with various analytical procedures for the determination of radiochemical purity of these preparations. (author)

  20. Rapid radiochemical separation of short-lived radionuclides in neutron-activated samples

    International Nuclear Information System (INIS)

    Fardy, J.

    1985-11-01

    Radiochemical separation procedures based on the removal of metal ions by columns of C 18 -bonded silica gel after selective complexation are examined and the simplicity of the method demonstrated by its application to determination of Mn, Cu and Zn in neutron-activated biological material from the following solutions (pH 0-10, sulphate concentration 0,18M and 1,44M SO 4 ): 8-hydroxyquinoline (oxine), ammonium pyrrolidinedithiocarbamate (APDC), cupferron (CUP), 1-(2-pyridylazo)-2-naphthol (PAN), 1-(2'-thiazolylazo)-2-naphthol (TAN), 4-(2-pyridylazo) resorcinol (PAR), diethylammonium diethyldithiocarbamate (DDC), potassium ethyl xanthate (PEX), acetylacetone (AcAc) or thenoyltrifluoracetone (TTA). The method is rapid and reliable and readily adaptable in all radiochemical laboratories

  1. Some problems concenrning the use of automated radiochemical separation systems in destructive neutron activation analysis

    International Nuclear Information System (INIS)

    Nagy, L.G.; Toeroek, G.

    1977-01-01

    The present state of a long term program is reviewed. It was started to elaborate a remote controlled automated radiochemical processing system for the neutron activation analysis of biological materials. The system is based on wet ashing of the sample followed by reactive desorption of some volatile components. The distillation residue is passed through a series of columns filled with selective ion screening materials to remove the matrix activity. The solution is thus ''stripped'' from the interfering radioions, and it is processed to single-elements through group separations using ion-exchange chromatographic techniques. Some special problems concerning this system are treated. (a) General aspects of the construction of a (semi)automated radiochemical processing system are discussed. (b) Comparison is made between various technical realizations of the same basic concept. (c) Some problems concerning the ''reconstruction'' of an already published processing system are outlined. (T.G.)

  2. Influence of EDC/NHS coupling chemistry on stability and cytotoxicity of ZnO nanoparticles modified with proteins

    Science.gov (United States)

    Keleştemur, Seda; Altunbek, Mine; Culha, Mustafa

    2017-05-01

    The toxicity of ZnO nanoparticles (NPs) is a growing concern due to its increasing use in several products including sunscreens, paints, pigments and ceramics for its antibacterial, antifungal, anti-corrosive and UV filtering properties. The toxicity of ZnO NPs is mostly attributed to the Zn2+ release causing an increase in the intracellular reactive oxygen species (ROS) level. The surface modification with a biocompatible ligand or a polymer can be a good strategy to reduce dissolution based toxicity. In two previous studies, the conflicting results with EDC/NHS coupling chemistry for ZnO NPs were reported. In this study, the same surface modification strategy with an emphasis on the stability of ZnO NPs is clarified. First, the density of -OH groups on the ZnO NPs is increased with hydrogen peroxide (H2O2) treatment, and then a silica coating on the ZnO NPs (Si-ZnO) surface is performed. Finally, a covalent attachment of bovine serum albumin (BSA) on three different concentrations of ZnO-Si is carried out by EDC/NHS coupling chemistry. ZnO NPs have a very high dissolution rate under acidic conditions of EDC/NHS coupling chemistry as determined from the ICP-MS analysis. In addition, the amount of ZnO NPs in coupling reaction has an important effect on the dissolution rate of Zn2+ and dependently BSA attached on the ZnO NP surfaces. Finally, the cytotoxicity of the BSA modified Si-ZnO NPs on human lung cancer (A549) and human skin fibroblast (HSF) is evaluated. Although an increased association of BSA modified ZnO NPs with cells was observed, the modification significantly decreased their cytotoxicity. This can be explained with the decreased active surface area of ZnO NPs with the surface modification. However, an increase in the mitochondrial depolarization and ROS production was observed depending on the amount of BSA coverage.

  3. Radiochemical separation of actinides for their determination in environmental samples and waste products

    Energy Technology Data Exchange (ETDEWEB)

    Gleisberg, B [Nuclear Engineering and Analytics Rossendorf, Inc. (VKTA), Dresden (Germany)

    1997-03-01

    The determination of low level activities of actinides in environmental samples and waste products makes high demands on radiochemical separation methods. Artificial and natural actinides were analyzed in samples form the surrounding areas of NPP and of uranium mines, incorporation samples, solutions containing radioactive fuel, solutions and solids resutling from the process, and in wastes. The activities are measured by {alpha}-spectrometry and {gamma}-spectrometry. (DG)

  4. Study of performance characteristics of a radiochemical method to determine uranium in biological samples

    International Nuclear Information System (INIS)

    Puga, Maria J.; Cerchietti, Maria L.R.; Prudenzo, J.E.; Arguelles, Maria G.

    2005-01-01

    In this paper is described a methodology to calculate detection limit (Ld), quantification level (Lq) and minimum detectable activity (MDA) in a radiochemical method for determination of uranium in urine samples. The concentration is measured by fluorimetry and alpha gross activity using liquid scintillation counting (LSC). The calculation of total propagated uncertainty on a spike sample is presented. Furthermore, the major sources of uncertainty and percentage contribution in both measurements are assessed. (author)

  5. A radiochemical technique for the determination of mercury in drinking water

    International Nuclear Information System (INIS)

    Ahmad, S.; Chaudhri, M.S.; Qureshi, I.H.

    1982-01-01

    A radiochemical method for the separation and determination of mercury in drinking water has been developed. The radionuclides of mercury formed after neutron irradiation of the sample were separated by liquid/liquid extraction using PAN and TBA mixture in chloroform from aqueous nitric acid medium. Quantitative extraction of mercury was achieved in a single step and the equilibrium was attained within five minutes. (orig.)

  6. Determination of mercury in biologycal samples by radiochemical neutron activation analysis

    International Nuclear Information System (INIS)

    Suc, N.V.

    1989-01-01

    The radiochemical neutron activation analysis was applied to determine contents of mercury in biological samples. Samples were digested in mixing of H 2 SO 4 and HNO 3 acid. After extraction of mercury by Ni-Ditiodietylphosphoric acid in carbontetrachloride, mercury was back extracted by 5% KI solution. Contents of mercury from five samples of fish was determined by this method. The accuracy of the method was checked by comparing it with NBS standard samples and results are good agreement

  7. Safety decommissioning regulations of radiochemical objects - the problem, requires urgent decision

    International Nuclear Information System (INIS)

    Rovnyj, S.I.; Arsent'eva, N.V.; Emel'yanov, N.M.; Kolesnikov, V.N.

    2001-01-01

    The necessity of planning and pursuance of the measures on decommissioning of radiochemical industry is discussed. Technological processes were stopped more than in 30 buildings and constructions of the PO Mayak. The characteristics of the technological buildings to be decommissioned were treated in the context of building peculiarities, function, character and level of contamination. An acceptable variant for reactor decommissioning invites development of the standard-legal aspects [ru

  8. Radiochemical determination and separation or total radium, 226Ra and 224Ra

    International Nuclear Information System (INIS)

    Suarez, J. A.; Gonzalez, J. A.; Pablo, M. A. de

    1987-01-01

    Radiochemical purification and separation of radium has been carried out and the determination of total radium solubilized in aqueous samples has been studied assuming that all the alpha emitters of the sample have their origin in the 226Ra and elements of its desintegration chain. Also, the activities of 22Ra and 226 Ra have been evaluated separately doing a measurement after the chemical separation of the radium and another one 10 days after. (Author) 9 refs

  9. Statistical analysis of radiochemical measurements of TRU radionuclides in REDC waste

    International Nuclear Information System (INIS)

    Beauchamp, J.; Downing, D.; Chapman, J.; Fedorov, V.; Nguyen, L.; Parks, C.; Schultz, F.; Yong, L.

    1996-10-01

    This report summarizes results of the study on the isotopic ratios of transuranium elements in waste from the Radiochemical Engineering Development Center actinide-processing streams. The knowledge of the isotopic ratios when combined with results of nondestructive assays, in particular with results of Active-Passive Neutron Examination Assay and Gamma Active Segmented Passive Assay, may lead to significant increase in precision of the determination of TRU elements contained in ORNL generated waste streams

  10. Analysis of the radiochemical purity of 18F-FDG by HPLC

    International Nuclear Information System (INIS)

    Chen Liguang; Tang Anwu; He Shanzhen; Chen Yulong

    2001-01-01

    The radiochemical purity (RCP) of 18 F-FDG is analyzed by HPLC. Eighty-five percent acetonitrile is used as the eluting solution. Carbon hydrate column is used as separation column. The t R of 18 F - is 6.50 min and 18 F-FDG is 9.00 min. HPLC take less time and has higher sensitivity than TLC for the same sample at the same time. So HPLC excels TLC in analyzing RCP of 18 F-FDG

  11. Genetically modified rice Bt-Shanyou63 expressing Cry1Ab/c protein does not harm Daphnia magna.

    Science.gov (United States)

    Zhang, Li; Guo, Ruqing; Fang, Zhixiang; Liu, Biao

    2016-10-01

    The genetically modified (GM) rice Bt-ShanYou63 (Bt-SY63) received an official biosafety certificate while its safety remained in dispute. In a lifelong study, Daphnia magna were experimentally fed a basal diet of rice flours from Bt-SY63 or its parental rice ShanYou63 (SY63) at concentrations of 0.2mg, 0.3mg, or 0.4mgC (per individual per day). Overall the survival, body size, and reproduction of the animals were comparable between Bt-SY63 and ShanYou63.. The results showed that no significant differences were observed in growth and reproduction parameters between D. magna fed GM and non-GM flour and no dose-related changes occurred in all the values. Based on the different parameters assessed, the GM rice Bt-SY63 is a safe food source for D. magna that does not differ in quality from non-GM rice. Copyright © 2016. Published by Elsevier Inc.

  12. A 90-day subchronic feeding study of genetically modified rice expressing Cry1Ab protein in Sprague-Dawley rats.

    Science.gov (United States)

    Song, Huan; He, Xiaoyun; Zou, Shiying; Zhang, Teng; Luo, Yunbo; Huang, Kunlun; Zhu, Zhen; Xu, Wentao

    2015-04-01

    Bacillus thuringiensis (Bt) transgenic rice line (mfb-MH86) expressing a synthetic cry1Ab gene can be protected against feeding damage from Lepidopteran insects, including Sesamia inferens, Chilo suppressalis, Tryporyza incertulas and Cnaphalocrocis medinalis. Rice flour from mfb-MH86 and its near-isogenic control MH86 was separately formulated into rodent diets at concentrations of 17.5, 35 and 70 % (w/w) for a 90-day feeding test with rats, and all of the diets were nutritionally balanced. In this study, the responses of rats fed diets containing mfb-MH86 were compared to those of rats fed flour from MH86. Overall health, body weight and food consumption were comparable between groups fed diets containing mfb-MH86 and MH86. Blood samples were collected prior to sacrifice and a few significant differences (p < 0.05) were observed in haematological and biochemical parameters between rats fed genetically modified (GM) and non-GM diets. However, the values of these parameters were within the normal ranges of values for rats of this age and sex, thus not considered treatment related. In addition, upon sacrifice a large number of organs were weighed, macroscopic and histopathological examinations were performed with only minor changes to report. In conclusion, these results demonstrated that no toxic effect was observed in the conditions of the experiment, based on the different parameters assessed. GM rice mfb-MH86 is as safe and nutritious as non-GM rice.

  13. Synthesis of N-[methyl-11C]hydromorphone by using multivariate strategies for optimization of radiochemical yields

    International Nuclear Information System (INIS)

    Rimland, Annika; Bergson, Goeran; Obenius, Ulf; Sjoeberg, Stefan; Langstroem, Bengt

    1987-01-01

    The synthesis of N-[methyl- 11 C]hydromorphone has been performed by using [ 11 C]methyl iodide and desmethyl hydromorphone in a mixture of dimethylsulphoxide and dimethylformamide as solvent. Optimization of the radiochemical yield by varying the reaction conditions was performed by using multivariate strategies. The labelled hydromorphone was obtained in 72% radiochemical yield in the alkylation reaction with [ 11 C]-methyl iodide, counted from the end of the [ 11 C]methyl iodide synthesis. N-[Methyl- 11 C]hydromorphone was obtained as a ready injectable pharmaceutical solution with a total synthesis time of 40 min and in a 10% total radiochemical yield, with a radiochemical purity > 99.5%, according to HPLC analysis. (author)

  14. Defining Optimized Properties of Modified mRNA to Enhance Virus- and DNA- Independent Protein Expression in Adult Stem Cells and Fibroblasts

    Directory of Open Access Journals (Sweden)

    Frauke Hausburg

    2015-02-01

    Full Text Available Background: By far, most strategies for cell reprogramming and gene therapy are based on the introduction of DNA after viral delivery. To avoid the high risks accompanying these goals, non-viral and DNA-free delivery methods for various cell types are required. Methods: Relying on an initially established PCR-based protocol for convenient template DNA production, we synthesized five differently modified EGFP mRNA (mmRNA species, incorporating various degrees of 5-methylcytidine-5'-triphosphate (5mC and pseudouridine-5'-triphosphate (Ψ. We then investigated their effect on i protein expression efficiencies and ii cell viability for human mesenchymal stem cells (hMSCs and fibroblasts from different origins. Results: Our protocol allows highly efficient mmRNA production in vitro, enabling rapid and stable protein expression after cell transfection. However, our results also demonstrate that the terminally optimal modification needs to be defined in pilot experiments for each particular cell type. Transferring our approach to the conversion of fibroblasts into skeletal myoblasts using mmRNA encoding MyoD, we confirm the huge potential of mmRNA based protein expression for virus- and DNA-free reprogramming strategies. Conclusion: The achieved high protein expression levels combined with good cell viability not only in fibroblasts but also in hMSCs provides a promising option for mmRNA based modification of various cell types including slowly proliferating adult stem cells. Therefore, we are confident that our findings will substantially contribute to the improvement of efficient cell reprogramming and gene therapy approaches.

  15. Radiochemical problems of radiation chemical synthesis in n, γ-field of nuclear reactor

    International Nuclear Information System (INIS)

    Mironov, V.P.; Frejdus, N.V.; Bugaenko, L.T.; Kalyazin, E.P.; Petryaev, E.P.

    1981-01-01

    A wide applicability of products of radiation chemical synthesis (RCS), using n, γ-irradiation, is limited by possible contamination of the latter with long-lived radioactive isotopes of chemical elements included in the composition of the reagent and compounds syntesized (chemically non-separable radionuclides - CNR). A technique of the determination of the limit accumulation CNR on the basis of radiation chemical parameters of the synthesis (radiation-chemical yield, the dose rate absorbed, singleness of purpose of RCS etc.) and radiochemical parameters of formation and accumulation of CNR (radiochemical yields of CNR in the products of radiolysis, neutron fluence, the reagent purity etc.) is suggested. The radiochemical evaluation of CNR accumulation (tritium and carbon-14), formed at the expense of activation with neutrons of chemical elements of water and organic substances, consisting of hydrogen, carbon and oxygen has shown that at relatively low yields of final products (> or approximately 3 molecules/100 eV) no accumulation of radionuclides in concentrations reaching the average admissible concentration takes place [ru

  16. Radiochemical aging of an epoxy network; Vieillissement radiochimique d'un reseau epoxyde

    Energy Technology Data Exchange (ETDEWEB)

    Devanne, Th

    2003-05-01

    This thesis is to give a better understanding of the radiochemical aging of a thermoset resin under gamma irradiation. The conditions of aging are gamma irradiation under air with a dose rate of 2 kGy/h at 120 C. The requested lifetime is four years, it means a dose of 70 MGy. The first step of this work was the choice of a resistive epoxy resin. This choice was made thanks to the literature data. The high glass transition temperature and the high amount of aromatic groups were the main criteria of the final choice. After this choice, thermal and mechanical properties were followed under thermal and radiochemical aging: i) under thermal aging, after 600 hours at 220 C, the glass transition temperature remained unchanged. But, from a mechanical point of view, properties at break dramatically decreased. This embrittlement was assigned to a critical oxidized layer. The thickness of this layer was estimated about 30 {mu}m. ii) the same kind of embrittlement was observed under radiochemical aging. Moreover, it appeared a decrease of the glass transition temperature when increasing the dose of irradiation. This indicates that the main degradation mechanism is chain scission under anaerobic atmosphere. We, then, proposed a mechanistic model associated with a kinetic model to predict the evolution of the glass transition temperature depending on the irradiation conditions. Parameters of the kinetic model were determined by solid NMR and ESR experiments. Comparison between experimental and calculated values at 120 C is satisfactory, a global good agreement was found. (author)

  17. Radiochemical determination of zirconium by inductively coupled plasma mass spectrometry (ICPMS)

    International Nuclear Information System (INIS)

    Oliveira, Thiago C.; Oliveira, Arno Heeren de

    2013-01-01

    The zirconium isotope 93 Zr is a long-lived pure β-particle-emitting radionuclide thus occurring as one of the radionuclides found in nuclear reactors. It's produced from 235 U fission and from 92 Zr neutron activation. Due to its long half-life, 93 Zr is one of the interest radionuclides for assessment studies performance of waste storage or disposal. Measurement of 93 Zr is difficult owing to its trace level concentration and its low activity in nuclear wastes and further because its certified standards are not frequently available. The aim of this work was to apply a selective radiochemical separation methodology for 93 Zr determination in nuclear waste and analyze it by Inductively Coupled Plasma Mass Spectrometry (ICPMS). To set up the zirconium radiochemical separation procedure, a zirconium tracer solution was used in order to follow the zirconium behavior during the radiochemical separation. A tracer solution containing the main interferences, Ba, Co, Eu, Fe, Mn, Nb, Ni, Sr, and Y was used in order to verify the decontamination factor during separation process. The limit of detection of 0,039 ppb was obtained for zirconium standard solutions by ICPMS. Then, the protocol will be applied to low level waste (LLW) and intermediate level waste (ILW) from nuclear power plants. (author)

  18. Influence of Storage Temperature on Radiochemical Purity of 99mTc-Radiopharmaceuticals.

    Science.gov (United States)

    Uccelli, Licia; Boschi, Alessandra; Martini, Petra; Cittanti, Corrado; Bertelli, Stefania; Bortolotti, Doretta; Govoni, Elena; Lodi, Luca; Romani, Simona; Zaccaria, Samanta; Zappaterra, Elisa; Farina, Donatella; Rizzo, Carlotta; Giganti, Melchiore; Bartolomei, Mirco

    2018-03-15

    The influence of effective room temperature on the radiochemical purity of 99m Tc-radiopharmaceuticals was reported. This study was born from the observation that in the isolators used for the preparation of the 99m Tc-radiopharmaceuticals the temperatures can be higher than those reported in the commercial illustrative leaflets of the kits. This is due, in particular, to the small size of the work area, the presence of instruments for heating, the continuous activation of air filtration, in addition to the fact that the environment of the isolator used for the 99m Tc-radiopharmaceuticals preparation and storage is completely isolated and not conditioned. A total of 244 99m Tc-radiopharmaceutical preparations (seven different types) have been tested and the radiochemical purity was checked at the end of preparation and until the expiry time. Moreover, we found that the mean temperature into the isolator was significantly higher than 25 °C, the temperature, in general, required for the preparation and storage of 99m Tc-radiopharmaceuticals. Results confirmed the radiochemical stability of radiopharmaceutical products. However, as required in the field of quality assurance, the impact that different conditions than those required by the manufacturer on the radiopharmaceuticals quality have to be verified before human administration.

  19. Investigation of quantitative separation of thorium, uranium, neptunium and plutonium from complex radiochemical mixtures

    International Nuclear Information System (INIS)

    Ushatskij, V.N.; Preobrazhenskaya, L.D.; Kolychev, V.B.; Gugel', E.S.

    1979-01-01

    Quantitative separation of actinides and their radiochemical purification with the aid of TBP with subsequent separation of thorium and quantitative separation of U, Np and Pu with the aid of D2EHPA have been studied. The method has been developed for quantitative extraction-chromatographic separation and radiochemical purification of nanogram amounts of U, Pu and microgram amounts of Th and Np from complex radiochemical mixtures containing both fragment radioisotopes and non-radioactive macrocomponents ( Fe,Al,Mg,Mn, Na and others). The method calls for application of one-extraction-chromatographic column with TBP and one column with D2EHPA. Thorium is separated at the first stage since it does not form complexes in a chloride solution during washing of the sorption column with 6. OM HCl. Npsup((4)) and Pusup((3)) required for separation are stabilized with the aid of hydrazine and hydroxylamine mixture. The yield of each of the above-cited actinide elements during the complete two-stage separation and at the stage of their separation varies within the range of 98.5-99.3%

  20. Spaceflight Activates Protein Kinase C Alpha Signaling and Modifies the Developmental Stage of Human Neonatal Cardiovascular Progenitor Cells.

    Science.gov (United States)

    Baio, Jonathan; Martinez, Aida F; Bailey, Leonard; Hasaniya, Nahidh; Pecaut, Michael J; Kearns-Jonker, Mary

    2018-02-12

    Spaceflight impacts cardiovascular function in astronauts; however, its impact on cardiac development and the stem cells that form the basis for cardiac repair is unknown. Accordingly, further research is needed to uncover the potential relevance of such changes to human health. Using simulated microgravity (SMG) generated by two-dimensional clinorotation and culture aboard the International Space Station (ISS), we assessed the effects of mechanical unloading on human neonatal cardiovascular progenitor cell (CPC) developmental properties and signaling. Following 6-7 days of SMG and 12 days of ISS culture, we analyzed changes in gene expression. Both environments induced the expression of genes that are typically associated with an earlier state of cardiovascular development. To understand the mechanism by which such changes occurred, we assessed the expression of mechanosensitive small RhoGTPases in SMG-cultured CPCs and observed decreased levels of RHOA and CDC42. Given the effect of these molecules on intracellular calcium levels, we evaluated changes in noncanonical Wnt/calcium signaling. After 6-7 days under SMG, CPCs exhibited elevated levels of WNT5A and PRKCA. Similarly, ISS-cultured CPCs exhibited elevated levels of calcium handling and signaling genes, which corresponded to protein kinase C alpha (PKCα), a calcium-dependent protein kinase, activation after 30 days. Akt was activated, whereas phosphorylated extracellular signal-regulated kinase levels were unchanged. To explore the effect of calcium induction in neonatal CPCs, we activated PKCα using hWnt5a treatment on Earth. Subsequently, early cardiovascular developmental marker levels were elevated. Transcripts induced by SMG and hWnt5a-treatment are expressed within the sinoatrial node, which may represent embryonic myocardium maintained in its primitive state. Calcium signaling is sensitive to mechanical unloading and directs CPC developmental properties. Further research both in space and on Earth

  1. Basal-bolus insulin therapy reduces maternal triglycerides in gestational diabetes without modifying cholesteryl ester transfer protein activity.

    Science.gov (United States)

    Olmos, Pablo R; Borzone, Gisella R

    2017-09-01

    Macrosomia in the offspring of overweight/obese mothers with glucose-controlled gestational diabetes mellitus (GDM) is due to excessive rise of maternal triglycerides (TG). We aimed to ascertain whether basal-bolus insulin therapy (BBIT), or other components of the treatment, could reduce TG in GDM. We studied the records of 131 singleton pregnancies with GDM, using stepwise multiple linear regression, Mann-Whitney, χ 2 , and Jonckheere-Terpstra tests. As maternal TG increased steadily during normal pregnancy, these were transformed as z-scores. The atherogenic index of plasma (AIP) was calculated as a measure of cholesteryl ester transfer protein activity. Multiple regression showed that only BBIT (but neither limitation of weight gain nor metformin) reduced maternal TG z-scores (P = 0.011). When the 131 pregnancies were split into two groups - without BBIT (n = 58; HbA1c = 5.3 ± 0.3%) and with BBIT (n = 73; HbA1c = 5.4 ± 0.6; P = 0.2005) - we observed that BBIT (n = 73) reduced maternal TG z-scores in a dose-related fashion (Jonckheere-Terpstra P = 0.03817). The atherogenic index of plasma remained within normal range in both groups. BBIT (but not weight gain control nor metformin) reduced maternal TG in mothers with glucose-controlled GDM. This beneficial effect of BBIT was not related to changes in the cholesteryl ester transfer protein activity. © 2017 Japan Society of Obstetrics and Gynecology.

  2. Effective intracellular inhibition of the cAMP-dependent protein kinase by microinjection of a modified form of the specific inhibitor peptide PKi in living fibroblasts.

    Science.gov (United States)

    Fernandez, A; Mery, J; Vandromme, M; Basset, M; Cavadore, J C; Lamb, N J

    1991-08-01

    In order to obtain a peptide retaining its biological activity following microinjection into living cells, we have modified a synthetic peptide [PKi(m)(6-24)], derived from the specific inhibitor protein of the cAMP-dependent protein kinase (A-kinase) in two ways: (1) substitution of the arginine at position 18 for a D-arginine; (2) blockade of the side chain on the C-terminal aspartic acid by a cyclohexyl ester group. In an in vitro assay, PKi(m) has retained a specific inhibitory activity against A-kinase as assessed against six other kinases, with similar efficiency to that of the unmodified PKi(5-24) peptide. Microinjection of PKi(m) into living fibroblasts reveals its capacity to prevent the changes in cell morphology and cytoskeleton induced by drugs which activate endogenous A-kinase, whereas the original PKi peptide failed to do so. This inhibition of A-kinase in vivo by PKi(m) lasts between 4 and 6 h after injection. In light of its effective half-life, this modified peptide opens a route for the use of biologically active peptides in vivo, an approach which has been hampered until now by the exceedingly short half-life of peptides inside living cells. By providing a direct means of inhibiting A-kinase activity for sufficiently long periods to observe effects on cellular functions in living cells, PKi(m) represents a powerful tool in studying the potential role of cAMP-dependent phosphorylation in vivo.

  3. [Alternation of proteins in brain of Parkinson's disease model rats after the transplantation of TH-NTN gene modified bone marrow mesenchymal stem cells].

    Science.gov (United States)

    Huang, Yue; Chang, Cheng; Zhang, Jie-wen; Gao, Xiao-qun

    2012-09-04

    To explore the effects of tyrosine hydroxylase-neurturin (TH-NTN) gene modified bone marrow mesenchymal stem cell (BMSC) transplantation in Parkinson's disease (PD) model rats and the alternations of correlated proteins. The PD rat model was established by the 2-point injection of 6-hydroxydopamine (6-OHDA) into unilateral (right) striatum. Successful modeling rats were separated into PD, BMSC and TH-NTN-BMSC groups. BMSC and TH-NTN-BMSC groups were transplanted into BMSCs and TH-NTN gene modified BMSC cells separately into right striatum. After transplantation, ethology detection in all groups was made with an intraperitoneal injection of apomorphine (APO). Dopamine (DA) and Dihydroxyphenylacetic Acid (DOPAC) in striatum were detected by high performance liquid electrochemical analysis. TH and NTN proteins in right striatum were also analyzed by immunohistochemistry and Western blot. Finally the density of dopamine receptors in post synaptic density of dopaminergic synapses of corpus striatum were compared between each group by post-embedding immunogold electron microscopy. After an injection of APO, rotation frequency decreased in TH-NTN-BMSC group, i.e. (5.7 ± 1.3) circles/min versus (10.8 ± 2.2), (9.9 ± 1.2) circles/min in PD and BMSC groups (P TH-NTN-BMSC group versus (923 ± 132)/µm(2) in PD and (860 ± 116)/µm(2) in BMSC groups was also found. The combined therapy of TH and NTN genes increases the synthesis of DA and also protects the dopaminergic neurons to achieve double therapeutic effects. It may provide potential innovations of PD genetic therapy.

  4. A 90-day safety study of genetically modified rice expressing rhIGF-1 protein in C57BL/6J rats.

    Science.gov (United States)

    Tang, Maoxue; Xie, Tingting; Cheng, Wenke; Qian, Lili; Yang, Shulin; Yang, Daichang; Cui, Wentao; Li, Kui

    2012-06-01

    Genetically modified plants expressing disease resistance traits offer new treatment strategies for human diseases, but at the same time present a challenge in terms of food safety assessment. The present 90-day feeding study was designed to assess the safety of transgenic rice expressing the recombinant human insulin-like growth factor-1 (rhIGF-1) compared to its parental wild rice. Male and female C57BL/6J rats were given a nutritionally balanced purified diet with 20% transgenic rhIGF-1 rice or 20% parental rice for 90 days. This corresponds to a mean daily rhIGF-1 protein intake of approximately 217.6 mg/kg body weight based on the average feed consumption. In the animal study a range of biological, biochemical, clinical, microbiological and pathological parameters were examined and several significant differences were observed between groups, but none of the effects were considered to be adverse. In conclusion, no adverse or toxic effects on C57BL/6J rats were observed in the design used in this 90-day study. These results will provide valuable information for the safety assessment of genetically modified food crops.

  5. Characterization of opaque2 modifier QTLs and candidate genes in recombinant inbred lines derived from the K0326Y quality protein maize inbred

    KAUST Repository

    Holding, David R.

    2010-11-13

    Quality protein maize (QPM) is a high lysine-containing corn that is based on genetic modification of the opaque2 (o2) mutant. In QPM, modifier genes convert the starchy endosperm of o2 to the vitreous phenotype of wild type maize. There are multiple, unlinked o2 modifier loci (Opm) in QPM and their nature and mode of action are unknown. We previously identified seven Opm QTLs and characterized 16 genes that are differentially up-regulated at a significant level in K0326Y QPM, compared to the starchy endosperm mutant W64Ao2. In order to further characterize these Opm QTLs and the genes up-regulated in K0326Y QPM, we created a population of 314 recombinant inbred lines (RILs) from a cross between K0326Y QPM and W64Ao2. The RILs were characterized for three traits associated with endosperm texture: vitreousness, density and hardness. Genetic linkage analysis of the RIL population confirmed three of the previously identified QTLs associated with o2 endosperm modification in K0326Y QPM. Many of the genes up-regulated in K0326Y QPM showed substantially higher levels of expression in vitreous compared with opaque RILs. These included genes associated with the upstream regulation of the ethylene response pathway, and a gene encoding a regulatory subunit of pyrophosphate-dependent fructose-6-phosphate 1-phosphotransferase, an adaptive enzyme of the glycolytic pathway. © 2010 Springer-Verlag.

  6. Coping strategy and social support modify the association between perceived stress and C-reactive protein: a longitudinal study of healthy men and women.

    Science.gov (United States)

    Shimanoe, Chisato; Hara, Megumi; Nishida, Yuichiro; Nanri, Hinako; Otsuka, Yasuko; Horita, Mikako; Yasukata, Jun; Miyoshi, Nobuyuki; Yamada, Yosuke; Higaki, Yasuki; Tanaka, Keitaro

    2018-05-01

    Inconsistent associations have been reported between perceived stress and C-reactive protein (CRP), a marker of systemic inflammation. We previously observed a male-specific inverse relationship between perceived stress and CRP in a cross-sectional study. In the present study, we examined the longitudinal association between changes in perceived stress and CRP, and further analyzed whether changes in coping strategies and social support modify this association. This study included 8454 participants in both a baseline survey and a follow-up survey 5 years later. Psychosocial measures (i.e. perceived stress, coping strategies, and social support) and CRP concentrations were measured by identical means in both surveys. Consistent with our previous findings, increased perceived stress was significantly associated with lower CRP in men (p trend  = .037), but not in women. Increased "emotional expression," a coping strategy, was also associated with lower CRP in women (p trend  = .024). Furthermore, interactions between perceived stress and a coping strategy (positive reappraisal) or social support on CRP were found in men (p interaction  = .007 and .038, respectively); the above inverse association between stress and CRP was not detected for participants with diminished positive reappraisal or social support. In conclusion, increases in perceived stress during a 5-year period were associated with decreases in CRP among healthy men, and the observed association was possibly modified by coping strategy or social support.

  7. Study of the interaction of 6-mercaptopurine with protein by microdialysis coupled with LC and electrochemical detection based on functionalized multi-wall carbon nanotubes modified electrode.

    Science.gov (United States)

    Cao, Xu-Ni; Lin, Li; Zhou, Yu-Yan; Zhang, Wen; Shi, Guo-Yue; Yamamoto, Katsunobu; Jin, Li-Tong

    2003-07-14

    Microdialysis sampling coupled with liquid chromatography and electrochemical detection (LC-ECD) was developed and applied to study the interaction of 6-Mercaptopurine (6-MP) with bovine serum albumin (BSA). In the LC-ECD, the multi-wall carbon nanotubes fuctionalized with carboxylic groups modified electrode (MWNT-COOH CME) was used as the working electrode for the determination of 6-MP. The results indicated that this chemically modified electrode (CME) exhibited efficiently electrocatalytic oxidation for 6-MP with relatively high sensitivity, stability and long-life. The peak currents of 6-MP were linear to its concentrations ranging from 4.0 x 10(-7) to 1.0 x 10(-4) mol l(-1) with the calculated detection limit (S/N = 3) of 2.0 x 10(-7) mol l(-1). The method had been successfully applied to assess the association constant (K) and the number of the binding sites (n) on a BSA molecular, which calculated by Scatchard equation, were 3.97 x 10(3) mol(-1) l and 1.51, respectively. This method provided a fast, sensible and simple technique for the study of drug-protein interactions.

  8. Modifying effects of 5-azacytidine on metal-containing proteins profile in Guerin carcinoma with different sensitivity to cytostatics.

    Science.gov (United States)

    Chekhun, V F; Lozovska, Y V; Naleskina, L A; Borikun, T V; Burlaka, A P; Todor, I N; Demash, D V; Yalovenko, T M; Zadvornyi, T V; Pavlova, A O; Storchay, D M; Lukianova, N Yu

    2016-12-01

    To assess the influence of the treatment with 5-azacytidine (5-aza) on the profile of metal-containing proteins and factors of their regulation in Guerin carcinoma cells in vivo. The study was conducted on Wistar rats transplanted with wild-type Guerin carcinoma (Guerin/WT) and its strains resistant to cisplatin (Guerin/CP) or doxorubicin (Guerin/Dox). Animals were distributed in 6 groups treated with 5-aza and control animals without treatment. 5-Aza was injected by i.v. route (1 injection in 4 days at a dose of 2 mg/kg starting from the 4 th day after tumor transplantation, 4 injections in total). Ferritin levels in blood serum and tumor tissue were measured by ELISA, transferrin and free iron complexes - by low-temperature EPR, miRNA-200b, -133a and -320a levels and promoter methylation - by real-time quantitative reverse transcription polymerase chain reaction. The study has shown that 5-aza treatment caused demethylation of promoter regions of fth1 and tfr1 genes in all studied Guerin carcinoma strains. 5-Aza treatment resulted in a significant decrease of ferritin levels in tumor tissue (by 32.1% in Guerin/WT strain, by 29.8% in Guerin/Dox and by 69.1% in Guerin/CP). These events were accompanied by 3.5-fold and 2-fold increase of free iron complexes levels in tumor tissue of doxorubicin and cisplatin resistant strains, respectively. Also, 5-aza treatment resulted in significantly elevated levels of miR-200b, -133a, 320a expression in tumor tissue. After 5-aza treatment, ferritin levels in blood serum of animals with Guerin/Dox were increased by 23.9%, while in Guerin/Wt and Guerin/CP they were decreased by 17 and 16%, respectively. Alterations of epigenetic regulation upon in vivo treatment with 5-aza change the levels of metal-containing proteins due to DNA demethylation and altered miRNA expression profiles in Guerin carcinoma cells.

  9. Ultra-sensitive chemiluminescent detection of Staphylococcus aureus based on competitive binding of Staphylococcus protein A-modified magnetic beads to immunoglobulin G

    International Nuclear Information System (INIS)

    Xiong, Jie; Wang, Wenwen; Zhou, Yali; Kong, Weijun; Wang, Zhenxing; Fu, Zhifeng

    2016-01-01

    Staphylococcus protein A (SPA) is a surface protein only expressed naturally in the cell walls of Staphylococcus aureus (S. aureus) and binds specifically to the Fc region of immunoglobulin G (IgG). This fact can be utilized for the detection of S. aureus. Specifically, SPA-modified magnetic beads, compete with S. aureus pathogens for binding to rabbit IgG that previously was labeled with horseradish peroxidase (HRP). The beads were then magnetically separated, and chemiluminescence (CL) was generated by adding the reagents luminol and H_2O_2. Under optimal conditions, the intensity of CL decreases with increasing concentration of S. aureus over a very wide linear range (10 to 1.0 × 10"9 cfu·mL"−"1), with a limit of detection of 6.0 cfu·mL"−"1 at an S/N ratio of 3. The assay (including binding reaction, magnetic separation, washing of beads and detection) is completed within 50 min which is faster than many reported methods. It can well distinguish S. aureus from other Gram-positive and Gram-negative bacteria. The magnetic beads have the beneficial effect of eliminating undesired matrix effects and of concentrating the sample. The method was applied to the analysis of urine, apple juice and glucose injection samples spiked with S. aureus, and recoveries ranged from 85 to 107 %. (author)

  10. Subchronic toxicity study in vivo and allergenicity study in vitro for genetically modified rice that expresses pharmaceutical protein (human serum albumin).

    Science.gov (United States)

    Sheng, Yao; Qi, Xiaozhe; Liu, Yifei; Guo, Mingzhang; Chen, Siyuan; He, Xiaoyun; Huang, Kunlun; Xu, Wentao

    2014-10-01

    Genetically modified (GM) crops that express pharmaceutical proteins have become an important focus of recent genetic engineering research. Food safety assessment is necessary for the commercial development of these crops. Subchronic toxicity study in vivo and allergenicity study in vitro were designed to evaluate the food safety of the rice variety expressing human serum albumin (HSA). Animals were fed rodent diets containing 12.5%, 25.0% and 50.0% GM or non-GM rice for 90 days. The composition analysis of the GM rice demonstrated several significant differences. However, most of the differences remained within the ranges reported in the literature. In the animal study, a range of indexes including clinical observation, feed efficiency, hematology, serum chemistry, organ weights and histopathology were examined. Random changes unrelated to the GM rice exposure, within the range of historical control values and not associated with any signs of illness were observed. The results of heat stability and in vitro digestion of HSA indicated no evidence of potential allergenicity of the protein. Overall, the results of these studies suggest that the GM rice appears to be safe as a dietary ingredient when it is used at up to 50% in the diet on a subchronic basis. Copyright © 2014 Elsevier Ltd. All rights reserved.

  11. Effects of a diet containing genetically modified rice expressing the Cry1Ab/1Ac protein (Bacillus thuringiensis toxin) on broiler chickens.

    Science.gov (United States)

    Li, Zeyang; Gao, Yang; Zhang, Minhong; Feng, Jinghai; Xiong, Yandan

    2015-01-01

    The aim of this study was to evaluate the effect of feeding Bacillus thuringiensis (Bt) rice expressing the Cry1Ab/1Ac protein on broiler chicken. The genetically modified (GM) Bt rice was compared with the corresponding non-GM rice regarding performance of feeding groups, their health status, relative organ weights, biochemical serum parameters and occurrence of Cry1Ab/1Ac gene fragments. One hundred and eighty day-old Arbor Acres female broilers with the same health condition were randomly allocated to the two treatments (6 replicate cages with 15 broilers in each cage per treatment). They received diets containing GM rice (GM group) or its parental non-GM rice (non-GM group) at 52-57% of the air-dried diet for 42 days. The results show that the transgenic rice had a similar nutrient composition as the non-GM rice and had no adverse effects on chicken growth, biochemical serum parameters and necropsy during the 42-day feeding period. In birds fed the GM rice, no transgenic gene fragments were detected in the samples of blood, liver, kidneys, spleen, jejunum, ileum, duodenum and muscle tissue. In conclusion, the results suggest that Bt rice expressing Cry1Ab/1Ac protein has no adverse effects on broiler chicken. Therefore, it can be considered as safe and used as feed source for broiler chicken.

  12. A Novel 1,4-Dihydropyridine Derivative Improves Spatial Learning and Memory and Modifies Brain Protein Expression in Wild Type and Transgenic APPSweDI Mice.

    Directory of Open Access Journals (Sweden)

    Baiba Jansone

    Full Text Available Ca2+ blockers, particularly those capable of crossing the blood-brain barrier (BBB, have been suggested as a possible treatment or disease modifying agents for neurodegenerative disorders, e.g., Alzheimer's disease. The present study investigated the effects of a novel 4-(N-dodecyl pyridinium group-containing 1,4-dihydropyridine derivative (AP-12 on cognition and synaptic protein expression in the brain. Treatment of AP-12 was investigated in wild type C57BL/6J mice and transgenic Alzheimer's disease model mice (Tg APPSweDI using behavioral tests and immunohistochemistry, as well as mass spectrometry to assess the blood-brain barrier (BBB penetration. The data demonstrated the ability of AP-12 to cross the BBB, improve spatial learning and memory in both mice strains, induce anxiolytic action in transgenic mice, and increase expression of hippocampal and cortical proteins (GAD67, Homer-1 related to synaptic plasticity. The compound AP-12 can be seen as a prototype molecule for use in the design of novel drugs useful to halt progression of clinical symptoms (more specifically, anxiety and decline in memory of neurodegenerative diseases, particularly Alzheimer's disease.

  13. In vitro digestibility of individual amino acids in rumen-undegraded protein: the modified three-step procedure and the immobilized digestive enzyme assay.

    Science.gov (United States)

    Boucher, S E; Calsamiglia, S; Parsons, C M; Stern, M D; Moreno, M Ruiz; Vázquez-Añón, M; Schwab, C G

    2009-08-01

    Three soybean meal, 3 SoyPlus (West Central Cooperative, Ralston, IA), 5 distillers dried grains with solubles, and 5 fish meal samples were used to evaluate the modified 3-step in vitro procedure (TSP) and the in vitro immobilized digestive enzyme assay (IDEA; Novus International Inc., St. Louis, MO) for estimating digestibility of AA in rumen-undegraded protein (RUP-AA). In a previous experiment, each sample was ruminally incubated in situ for 16 h, and in vivo digestibility of AA in the intact samples and in the rumen-undegraded residues (RUR) was obtained for all samples using the precision-fed cecectomized rooster assay. For the modified TSP, 5 g of RUR was weighed into polyester bags, which were then heat-sealed and placed into Daisy(II) incubator bottles. Samples were incubated in a pepsin/HCl solution followed by incubation in a pancreatin solution. After this incubation, residues remaining in the bags were analyzed for AA, and digestibility of RUP-AA was calculated based on disappearance from the bags. In vitro RUP-AA digestibility estimates obtained with this procedure were highly correlated to in vivo estimates. Corresponding intact feeds were also analyzed via the pepsin/pancreatin steps of the modified TSP. In vitro estimates of AA digestibility of the feeds were highly correlated to in vivo RUP-AA digestibility, which suggests that the feeds may not need to be ruminally incubated before determining RUP-AA digestibility in vitro. The RUR were also analyzed via the IDEA kits. The IDEA values of the RUR were good predictors of RUP-AA digestibility in soybean meal, SoyPlus, and distillers dried grains with solubles, but the IDEA values were not as good predictors of RUP-AA digestibility in fish meal. However, the IDEA values of intact feed samples were also determined and were highly correlated to in vivo RUP-AA digestibility for all feed types, suggesting that the IDEA value of intact feeds may be a better predictor of RUP-AA digestibility than the IDEA

  14. Silencing of OSBP-related protein 8 (ORP8) modifies the macrophage transcriptome, nucleoporin p62 distribution, and migration capacity

    International Nuclear Information System (INIS)

    Béaslas, Olivier; Vihervaara, Terhi; Li, Jiwei; Laurila, Pirkka-Pekka; Yan, Daoguang; Olkkonen, Vesa M.

    2012-01-01

    ORP8 is an oxysterol/cholesterol binding protein anchored to the endoplasmic reticulum and the nuclear envelope, and is abundantly expressed in the macrophage. We created and characterized mouse RAW264.7 macrophages with ORP8 stably silenced using shRNA lentiviruses. A microarray transcriptome and gene ontology pathway analysis revealed significant alterations in several nuclear pathways and ones associated with centrosome and microtubule organization. ORP8 knockdown resulted in increased expression and altered subcellular distribution of an interaction partner of ORP8, nucleoporin NUP62, with an intranuclear localization aspect and association with cytoplasmic vesicular structures and lamellipodial edges of the cells. Moreover, ORP8 silenced cells displayed enhanced migration, and a more pronounced microtubule cytoskeleton than controls expressing a non-targeting shRNA. ORP8 was shown to compete with Exo70 for interaction with NUP62, and NUP62 knockdown abolished the migration enhancement of ORP8-silenced cells, suggesting that the endogenous ORP8 suppresses migration via binding to NUP62. As a conclusion, the present study reveals new, unexpected aspects of ORP8 function in macrophages not directly involving lipid metabolism, but rather associated with nuclear functions, microtubule organization, and migration capacity. -- Highlights: ► The phenotype of Raw264.7 macrophage with ORP8 silenced is characterized. ► ORP8 silencing alters mRNA levels of nuclear and microtubule/centrosome pathways. ► ORP8 silencing results in increased expression and altered distribution of NUP62. ► ORP8 silenced macrophages show enhanced migration and altered microtubule cytoskeleton. ► ORP8 competes in vitro with Exo70 for binding to NUP62.

  15. Silencing of OSBP-related protein 8 (ORP8) modifies the macrophage transcriptome, nucleoporin p62 distribution, and migration capacity

    Energy Technology Data Exchange (ETDEWEB)

    Beaslas, Olivier; Vihervaara, Terhi [Minerva Foundation Institute for Medical Research, FI-00290 Helsinki (Finland); Li, Jiwei [Department of Biology, Jinan University, Guangzhou 510632 (China); Laurila, Pirkka-Pekka [FIMM, Institute for Molecular Medicine Finland, FI-00290 Helsinki (Finland); National Institute for Health and Welfare, Public Health Genomics Unit, FI-00290 Helsinki (Finland); Yan, Daoguang [Department of Biology, Jinan University, Guangzhou 510632 (China); Olkkonen, Vesa M., E-mail: vesa.olkkonen@helsinki.fi [Minerva Foundation Institute for Medical Research, FI-00290 Helsinki (Finland); Institute of Biomedicine, Anatomy, University of Helsinki, FI-00014 (Finland)

    2012-09-10

    ORP8 is an oxysterol/cholesterol binding protein anchored to the endoplasmic reticulum and the nuclear envelope, and is abundantly expressed in the macrophage. We created and characterized mouse RAW264.7 macrophages with ORP8 stably silenced using shRNA lentiviruses. A microarray transcriptome and gene ontology pathway analysis revealed significant alterations in several nuclear pathways and ones associated with centrosome and microtubule organization. ORP8 knockdown resulted in increased expression and altered subcellular distribution of an interaction partner of ORP8, nucleoporin NUP62, with an intranuclear localization aspect and association with cytoplasmic vesicular structures and lamellipodial edges of the cells. Moreover, ORP8 silenced cells displayed enhanced migration, and a more pronounced microtubule cytoskeleton than controls expressing a non-targeting shRNA. ORP8 was shown to compete with Exo70 for interaction with NUP62, and NUP62 knockdown abolished the migration enhancement of ORP8-silenced cells, suggesting that the endogenous ORP8 suppresses migration via binding to NUP62. As a conclusion, the present study reveals new, unexpected aspects of ORP8 function in macrophages not directly involving lipid metabolism, but rather associated with nuclear functions, microtubule organization, and migration capacity. -- Highlights: Black-Right-Pointing-Pointer The phenotype of Raw264.7 macrophage with ORP8 silenced is characterized. Black-Right-Pointing-Pointer ORP8 silencing alters mRNA levels of nuclear and microtubule/centrosome pathways. Black-Right-Pointing-Pointer ORP8 silencing results in increased expression and altered distribution of NUP62. Black-Right-Pointing-Pointer ORP8 silenced macrophages show enhanced migration and altered microtubule cytoskeleton. Black-Right-Pointing-Pointer ORP8 competes in vitro with Exo70 for binding to NUP62.

  16. Rapamycin and chloroquine: the in vitro and in vivo effects of autophagy-modifying drugs show promising results in valosin containing protein multisystem proteinopathy.

    Directory of Open Access Journals (Sweden)

    Angèle Nalbandian

    Full Text Available Mutations in the valosin containing protein (VCP gene cause hereditary Inclusion body myopathy (hIBM associated with Paget disease of bone (PDB, frontotemporal dementia (FTD, more recently termed multisystem proteinopathy (MSP. Affected individuals exhibit scapular winging and die from progressive muscle weakness, and cardiac and respiratory failure, typically in their 40s to 50s. Histologically, patients show the presence of rimmed vacuoles and TAR DNA-binding protein 43 (TDP-43-positive large ubiquitinated inclusion bodies in the muscles. We have generated a VCPR155H/+ mouse model which recapitulates the disease phenotype and impaired autophagy typically observed in patients with VCP disease. Autophagy-modifying agents, such as rapamycin and chloroquine, at pharmacological doses have previously shown to alter the autophagic flux. Herein, we report results of administration of rapamycin, a specific inhibitor of the mechanistic target of rapamycin (mTOR signaling pathway, and chloroquine, a lysosomal inhibitor which reverses autophagy by accumulating in lysosomes, responsible for blocking autophagy in 20-month old VCPR155H/+ mice. Rapamycin-treated mice demonstrated significant improvement in muscle performance, quadriceps histological analysis, and rescue of ubiquitin, and TDP-43 pathology and defective autophagy as indicated by decreased protein expression levels of LC3-I/II, p62/SQSTM1, optineurin and inhibiting the mTORC1 substrates. Conversely, chloroquine-treated VCPR155H/+ mice revealed progressive muscle weakness, cytoplasmic accumulation of TDP-43, ubiquitin-positive inclusion bodies and increased LC3-I/II, p62/SQSTM1, and optineurin expression levels. Our in vitro patient myoblasts studies treated with rapamycin demonstrated an overall improvement in the autophagy markers. Targeting the mTOR pathway ameliorates an increasing list of disorders, and these findings suggest that VCP disease and related neurodegenerative multisystem

  17. Molecular Properties of Guar Gum and Pectin Modify Cecal Bile Acids, Microbiota, and Plasma Lipopolysaccharide-Binding Protein in Rats.

    Science.gov (United States)

    Ghaffarzadegan, Tannaz; Marungruang, Nittaya; Fåk, Frida; Nyman, Margareta

    2016-01-01

    Bile acids (BAs) act as signaling molecules in various physiological processes, and are related to colonic microbiota composition as well as to different types of dietary fat and fiber. This study investigated whether guar gum and pectin-two fibers with distinct functional characteristics-affect BA profiles, microbiota composition, and gut metabolites in rats. Low- (LM) or high-methoxylated (HM) pectin, and low-, medium-, or high-molecular-weight (MW) guar gum were administered to rats that were fed either low- or high-fat diets. Cecal BAs, short-chain fatty acids (SCFA) and microbiota composition, and plasma lipopolysaccharide-binding protein (LBP) levels were analyzed, by using novel methodologies based on gas chromatography (BAs and SCFAs) and 16S rRNA gene sequencing on the Illumina MiSeq platform. Strong correlations were observed between cecal BA and SCFA levels, microbiota composition, and portal plasma LBP levels in rats on a high-fat diet. Notably, guar gum consumption with medium-MW increased the cecal amounts of cholic-, chenodeoxycholic-, and ursodeoxycholic acids as well as α-, β-, and ω-muricholic acids to a greater extent than other types of guar gum or the fiber-free control diet. In contrast, the amounts of cecal deoxycholic- and hyodeoxycholic acid were reduced with all types of guar gum independent of chain length. Differences in BA composition between pectin groups were less obvious, but cecal levels of α- and ω-muricholic acids were higher in rats fed LM as compared to HM pectin or the control diet. The inflammatory marker LBP was downregulated in rats fed medium-MW guar gum and HM pectin; these two fibers decreased the cecal abundance of Oscillospira and an unclassified genus in Ruminococcaceae, and increased that of an unclassified family in RF32. These results indicate that the molecular properties of guar gum and pectin are important for their ability to modulate cecal BA formation, gut microbiota composition, and high-fat diet induced

  18. An autoantibody against N{sup {epsilon}}-(carboxyethyl)lysine (CEL): Possible involvement in the removal of CEL-modified proteins by macrophages

    Energy Technology Data Exchange (ETDEWEB)

    Mera, Katsumi [Department of Biopharmaceutics, Graduate School of Pharmaceutical Sciences, Kumamoto University, Kumamoto (Japan); Nagai, Ryoji, E-mail: nagai-883@umin.ac.jp [Department of Food and Nutrition, Laboratory of Nutritional Science and Biochemistry, Japan Women' s University, Tokyo (Japan); Takeo, Kazuhiro; Izumi, Miyoko [Department of Biopharmaceutics, Graduate School of Pharmaceutical Sciences, Kumamoto University, Kumamoto (Japan); Maruyama, Toru [Department of Biopharmaceutics, Graduate School of Pharmaceutical Sciences, Kumamoto University, Kumamoto (Japan); Center for Clinical Pharmaceutical Science, Kumamoto University, Kumamoto (Japan); Otagiri, Masaki [Department of Biopharmaceutics, Graduate School of Pharmaceutical Sciences, Kumamoto University, Kumamoto (Japan); Faculty of Pharmaceutical Sciences, Sojo University, Kumamoto (Japan)

    2011-04-08

    Highlights: {yields} A higher amount of autoantibody against CEL than that of other AGEs was observed in human plasma. {yields} The purified human anti-CEL autoantibody specifically reacted with CEL. {yields} Anti-CEL antibody accelerated the uptake of {sup 125}I-CEL-HSA by macrophage in vitro. {yields} Endocytic uptake of {sup 125}I-CEL-HSA by mice liver was accelerated in the presence of anti-CEL antibody. -- Abstract: Advanced glycation end products (AGEs) are believed to play a significant role in the development of diabetic complications. In this study, we measured the levels of autoantibodies against several AGE structures in healthy human plasma and investigated the physiological role of the autoantibodies. A high titer of the autoantibody against N{sup {epsilon}}-(carboxyethyl)lysine (CEL) was detected in human plasma compared with other AGE structures such as CML and pentosidine. The purified human anti-CEL autoantibody reacted with CEL-modified human serum albumin (CEL-HSA), but not CML-HSA. A rabbit polyclonal anti-CEL antibody, used as a model autoantibody against CEL, accelerated the uptake of CEL-HSA by macrophages, but did not enhance the uptake of native HSA. Furthermore, when {sup 125}I-labeled CEL-HSA was injected into the tail vein of mice, accumulation of {sup 125}I-CEL-HSA in the liver was accelerated by co-injection of the rabbit anti-CEL antibody. These results demonstrate that the autoantibody against CEL in plasma may play a role in the macrophage uptake of CEL-modified proteins.

  19. A combined simple bubbling method with high performance liquid chromatography purification strategy, higher radiochemical yield and purity and faster preparation of carbon-11-raclopride.

    Science.gov (United States)

    Huang, Huacheng; Ning, Yanli; Zhang, Bucheng; Lou, Cen

    2015-01-01

    Carbon-11-raclopride (¹¹C-R) is a positron-emitting radiotracer successfully used for the study of cognitive control and widely applied in PET imaging. A simple automated preparation of ¹¹C-R by using the reaction of carbon-(11)-methyl triflate (¹¹C-MeOTF) or ¹¹C-methyl iodide (¹¹C-MeI) with demethylraclopride is described. Specifically we used a simple setup applied an additional "U" reaction vessel for ¹¹C-MeOTf compared with ¹¹C-MeI and assessed the influence of several solvents and of the amount of the percussor for ¹¹C-methylation of demethylraclopride by the bubbling method. The reversal of retention order between product and its precursor has been achieved for ¹¹C-R, enabling collection of the purified ¹¹C-R by using the HPLC column after shorter retention time. By the improved radiosynthesis and purification strategy, ¹¹C-R could be prepared with higher radiochemical yield than that of the previous studies. The yield for ¹¹C-MeOTf was 76% and for ¹¹C-CH3I >26% and with better radiochemical purity (>99% based on both ¹¹C-MeOTf and ¹¹C-MeI) as compared to the previously obtained purity of ¹¹C-R using HPLC method with acetonitrile as a part of mobile phase. Furthermore, by using ethanol as the organic modifier, residual solvent analysis prior to human injection could be avoided and ¹¹C-R could be injected directly following simple dilution and sterile filtration. Improved radiosynthesis and HPLC purification in combination with ethanol containing eluent, extremely shortened the time for preparation of ¹¹C-R, gave a higher radiochemical yield and purity for ¹¹C-R and can be used for multiple and faster synthesis of ¹¹C-R and probably for other ¹¹C-labeled radiopharmaceuticals.

  20. Preparation, radiochemical purity control and stability of 99mTc-mertiatide (Mag-3)

    International Nuclear Information System (INIS)

    Van Hemert, F.J.; Schimmel, K.J.M.; Van Eck-Smit, B.L.F.; Van Lenthe, H.

    2005-01-01

    Scintigraphic image analysis of 99m Tc-mertiatide (Mag-3, mercaptoacetyltriglycine) clearance provides the determination of the blood flow, the tubular transit time and the excretion as well from both kidneys. Radiopharmaceutical routine recommends a radiochemical purity control before administration of the product to a patient. The main objective of this study is to develop a Mag-3 labeling procedure that fits better than the previous one in our daily routine production of radiopharmaceuticals. Increasing proportions of 99m Tc-Mag-3 were measured during the heating and cooling steps of the Mag-3 labeling procedure. High performance liquid chromatography (HPLC) analysis was used to confirm the results of a rapid radiochemical quality control assay on standard instant thin-layer chromatography-silica gel (ITLC-SG) paper. The reconstitution time takes 20-25 minutes from the harvest of pertechnetate to a ready-for-use calibrated patient syringe. The HPLC profile of 99m Tc-Mag-3 including its minor impurities remains unchanged for 24-48 hours after reconstitution. The application of a programmable Peltier-directed device for heating/cooling provides a better control of the temperature course. The procedure proposed fully meets the labeling criteria recommended by the supplier and can be performed with a minimum of attention within a time-span that we formerly needed for solely the radiochemical purity control assay. Moreover, 99m Tc-Mag-3 prepared in this way seems to be considerably more stable than mentioned in the manufacturer's instructions. (author)

  1. Radiochemical determination of 210 Pb and 226Ra in petroleum sludges and scales

    International Nuclear Information System (INIS)

    Araujo, Andressa Arruda de

    2005-01-01

    The oil extraction and production, both onshore and offshore, can generate different types of residues, such as sludge, that is deposited in the water/oil separators, valves and storage tanks and scales, which form i the inner surface of ducts and equipment. Analyses already carried out through gamma spectrometry indicated the existence of high radioisotope concentration. However, radionuclides emitting low-energy gamma-rays, such as 210 Pb, are hardly detected by that technique. Consequently, there is a need to test alternative techniques to determine this and other radionuclides from the 238 U series. This work, therefore, focuses on the radiochemical determination of the concentration of 210 Pb, and 226 Ra in samples of sludge and scale from the oil processing stations of the UN-SEAL, a PETROBRAS unit responsible for the exploration and production of petroleum in Sergipe and Alagoas. The sludge and scale samples went through a preliminary process of extraction of oil, in order to separate the solid phase, where the largest fraction of the radioactivity is concentrated. After oil removal, the samples were digested using alkaline fusion as an option for dissolution. Finally, their activity concentration was determined for the samples of sludge and scales, using and alternative radiochemical method, which is based on ionic exchange. The activity concentration found for 210 Pb varied from 1,14 to 507,3 kBq kg -1 . The values for 226 Ra were higher, varying from 4,36 to 3.445 kBq kg -1 . The results for 226 Ra were then compared with the ones found for the same samples of sludge and scales using gamma spectrometry. The results of the comparison confirm the efficiency of the methodology used int hi work, that is, radiochemical determination by means of ionic exchange. (author)

  2. The radiochemical purity of technetium-99m-tin-diethylene-triamino-pentaacetic acid (DTPA) complex

    International Nuclear Information System (INIS)

    Besnard, M.; Costerousse, O.; Merlin, L.; Coehn, Y.

    1975-01-01

    The effect on radiochemical purity was studied as a function of the storage period of tin-DTPA solution and of the technetium-complex solution. The quantity of the pertechnetate ions present in the solution is determined by ascending paper chromatography, and an attempt was made to clarify the bond type of technetium by a spectrophotometric method. The tin-DTPA solutions for complexing of the reduced technetium are stable over a period of 8 weeks. The yield of the radiopharmaceutical product is better than 95%. (G.Gy.)

  3. Radiochemical and thermal studies of the cation-exchanged forms of synthetic zeolite Linde sieve A

    Energy Technology Data Exchange (ETDEWEB)

    Banerjee, S P [Saugar Univ. (India). Dept. of Chemistry

    1976-02-01

    The compositions of the cobalt and silver-exchanged forms of synthetic zeolite Sieve A have been determined by radiochemical and TGA studies and correspond to Co/sub 6/A.19.8H/sub 2/O and Ag/sub 12/..cap alpha... 20H/sub 2/O respectively (A=Al/sub 12/Si/sub 12/O/sub 48//sup 12/-). Heating of these zeolites inhibits their capacity for cation exchange and water absorption. No evidence of occluded NaAlO/sub 2/ has been found.

  4. Trace element evaluation of different varieties of chewing gum by radiochemical neutron activation analysis

    International Nuclear Information System (INIS)

    Zaidi, J.H.; Arif, M.; Fatima, I.; Ahmad, S.; Qureshi, I.H.

    2000-01-01

    Extensive use of chewing gums, by children in particular, entails the evaluation of trace element contents in them. Radiochemical neutron activation analysis (RNAA) was successfully employed to determine the concentration of 35 trace elements (essential, toxic and nonessential) in eight different brands of chewing gum generally consumed in Rawalpindi/Islamabad area. Comparison of trace element data of our work with literature has been presented. None of the elements detected in the brands of chewing gum examined was found to be present at a level representing a substantial contribution to the total dietary intake of the element. (author)

  5. Design of the Laboratory-Scale Plutonium Oxide Processing Unit in the Radiochemical Processing Laboratory

    Energy Technology Data Exchange (ETDEWEB)

    Lumetta, Gregg J. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Meier, David E. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Tingey, Joel M. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Casella, Amanda J. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Delegard, Calvin H. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Edwards, Matthew K. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Orton, Robert D. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Rapko, Brian M. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Smart, John E. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States)

    2015-05-01

    This report describes a design for a laboratory-scale capability to produce plutonium oxide (PuO2) for use in identifying and validating nuclear forensics signatures associated with plutonium production, as well as for use as exercise and reference materials. This capability will be located in the Radiochemical Processing Laboratory at the Pacific Northwest National Laboratory. The key unit operations are described, including PuO2 dissolution, purification of the Pu by ion exchange, precipitation, and re-conversion to PuO2 by calcination.

  6. Conceptual Design for the Pilot-Scale Plutonium Oxide Processing Unit in the Radiochemical Processing Laboratory

    Energy Technology Data Exchange (ETDEWEB)

    Lumetta, Gregg J.; Meier, David E.; Tingey, Joel M.; Casella, Amanda J.; Delegard, Calvin H.; Edwards, Matthew K.; Jones, Susan A.; Rapko, Brian M.

    2014-08-05

    This report describes a conceptual design for a pilot-scale capability to produce plutonium oxide for use as exercise and reference materials, and for use in identifying and validating nuclear forensics signatures associated with plutonium production. This capability is referred to as the Pilot-scale Plutonium oxide Processing Unit (P3U), and it will be located in the Radiochemical Processing Laboratory at the Pacific Northwest National Laboratory. The key unit operations are described, including plutonium dioxide (PuO2) dissolution, purification of the Pu by ion exchange, precipitation, and conversion to oxide by calcination.

  7. A new radiochemical assay for fructose-1,6-diphosphatase in human leucocytes

    International Nuclear Information System (INIS)

    Janssen, A.J.M.; Trijbels, F.J.M.

    1982-01-01

    Fructose-1,6-diphosphatase (D-fructose-1,6-diphosphate 1-phosphohydrolase, EC 3.1.3.11, FDPase) is one of the key enzymes of the gluconeogenic pathway. Measuring the activity both in the presence and in the absence of AMP yields the true FDPase activity, corrected for non-specific phosphatase activity. In this paper the authors introduce a new radiochemical assay for FDPase, based on the decarboxylating activity of 6-phosphogluconate dehydrogenase. One molecule [U- 14 C]fructose-1,6-diphosphate yields one molecule 14 CO 2 which can be captured in strongly basic solutions and counted in a liquid scintillation counter. (Auth.)

  8. Radiochemical procedure for the determination of plutonium isotopes in powdered milk

    International Nuclear Information System (INIS)

    Taddei, M.H.T.; Silva, N.C.

    2006-01-01

    A radiochemical procedure for the determination of alpha-emitting isotopes of plutonium in powdered milk is proposed. The procedure involves sample dissolution (by HNO 3 and HClO 4 ), separation by ionic-exchange resin, electrodeposition and alpha-spectroscopy. In order to determine the chemical recovery, 242 Pu was employed as a tracer. A reference material (Marine Sediment IAEA 135) was analyzed to validate such procedure, and to show its reliability. Afterwards, some powdered milk, produced for international trade, was analyzed and chemical recovery was found to be around 95%. (author)

  9. Methods for nuclear material control used in the basic production of a typical radiochemical plant

    International Nuclear Information System (INIS)

    Kositsyn, V.F.; Mukhortov, N.F.; Korovin, Yu.I.; Rudenko, V.S.; Petrov, A.M.

    1999-01-01

    Techniques for destructive and non-destructive assay of the component and isotopic composition of nuclear materials are described, namely gravimetric, titrimetric, coulometric, mass spectrometry, as well as those based on registration of neutron and γ radiations. Their metrologic characteristics are described. The techniques described are suggested to be used for nuclear material (NM) control and accounting purposes at the model radiochemical plant for processing irradiated fuel subassemblies from power reactors. The measurement control program is also described. This program is intended for the measurement quality assurance in the framework of NM control and accountancy system [ru

  10. Determination of trace elements in bottled water in Greece by instrumental and radiochemical neutron activation analyses

    International Nuclear Information System (INIS)

    Soupioni, M.J.; Symeopoulos, B.D.; Papaefthymiou, H.V.

    2006-01-01

    Four different bottled water brands sold in Greece in the winter of 2001-2002 were analyzed for a wide range of chemical elements, using neutron activation analysis (NAA). The elements Na and Br were determined instrumentally (INAA), whereas the other metals and trace elements radiochemically (RNAA). The results indicated that the mean level of all the elements determined in the samples were well within the European Union (EU) directive on drinking water and accomplish the drinking water standards of the World Health Organisation (WHO) as well as of the Food and Drug Administration (FDA). (author)

  11. Radiochemical techniques for determining some naturally occurring radionuclides in marine environmental materials

    Energy Technology Data Exchange (ETDEWEB)

    Baker, C W [Ministry of Agriculture, Fisheries and Food, Lowestoft (UK). Fisheries Lab.

    1984-06-15

    The determination of some of the naturally-occurring, alpha-emitting radionuclides in marine environmental materials, is of interest for several reasons. Radium and radon nuclides are potentially useful as oceanographic tracers. Lead and thorium nuclides may be used to study sedimentation rates, mixing processes and bioturbation in sediments. Radium and polonium nuclides are incorporated into food chains and the data may provide a perspective against which to assess the significance, for marine organisms, of exposure to radiation in a marine radioactive waste disposal situation. This paper discusses the manner in which samples are taken, and the radiochemical methods which have been employed to measure the nuclides, together with some data produced.

  12. Integral method of treatment of experimental data from radiochemical solar neutrino detectors

    International Nuclear Information System (INIS)

    Gavrin, V.N.; Kopylov, A.V.; Streltsov, A.V.

    1985-01-01

    An analysis is made of the statistical errors in solar neutrino detection by radiochemical detectors at different times of exposure. It is shown that short exposures (tau/sub e/ = one-half to one half-life) give minimal one-year error. The possibility is considered of the detection of the solar neutrino flux variation due to annual changes of the Earth-Sun distance. The integral method of treatment of the experimental data is described. Results are given of the statistical treatment of computer simulated data

  13. Radiochemical analysis of radio-nuclides in sea water collected near Bikini Atoll

    Energy Technology Data Exchange (ETDEWEB)

    Miyake, Y; Sugiura, Y

    1955-01-01

    A radiochemical analysis of sea water containing fission materials collected near Bikini Atoll in June, 1954, was performed. The sea water was boiled with hydrochloric acid, iron and lanthanum salts each 5 mg as Fe and La were added to it. They were precipitated as hydroxide, which was dissolved in hydrochloric acid and ferric chloride was extracted with ethyl ether. The remaining solution was evaporated to dryness and the residue was dissolved in hydrochloric acid. Using the latter solution the group separation was done with cation exchanger resins.

  14. Rapid and accurate determination of radiochemical purity of sup(99m)Tc compounds

    International Nuclear Information System (INIS)

    Tamat, S.R.

    1977-01-01

    The wide spread use of sup(99m)Tc-labelled radiopharmaceuticals and limitation of the short half-life of the isotope, is associated with an urgent need for a rapid, simple but accurate method for determining the radiochemical purity of the compound. A short paper chromatographic (KK) or thin layer chromatographic (KLT) method using 95% methanol or 0.9% saline solution as solvents, has solved the problem. With these methods, the amount of free sup(99m)Tc pertechnetate in a compound, can be determined in only a few minutes. These methods compare satisfactorily with lengtheir procedures. (author)

  15. The platinum group elements and gold: analysis by radiochemical and instrumental neutron activation analysis and relevance to geological exploration and related problems

    Energy Technology Data Exchange (ETDEWEB)

    Reeves, S.; Plimer, I. R. [Melbourne Univ., Parkville, VIC (Australia). School of Physics

    1996-12-31

    This paper presents an overview of research conducted with the support of the Australian Institute of Nuclear Science and Engineering, at the University of Melbourne, School of Earth Sciences, Radiochemical Neutron Activation Laboratory. The primary objective of this research is to realize the high potential of the platinum group elements (PGE) and gold to the solution of petrogenetic problems, the study of magma generation and magmatic processes in mafic/ultramafic rock suites, as tracers in hydrothermal ore formation. The PGEs (Os, Ru, Ir, Pt, Pd and Rh) are among the least abundant of all elements on earth with unique properties such as high melting points, high electrical and thermal conductivity, high density, strength and toughness as alloys. They exhibit both siderophile and chalcophile characteristics and are valuable tools in providing information about magmatic processes, in particular S-saturation, as well as crystal fractionation trends. Two distinct groups of PGEs are discerned; the IPGEs (Ru, Os, Ir) and the PPGEs (Pt, Pd, Rh, Au) on the basis of their behaviour during fractionation processes. Using chondrite normalized PGE patterns it is possible to distinguish between sulphides that segregated from primitive magmas, such as komatiites, and sulphides which segregated from more fractionated magmas, such as tholeiites. It is critical to the understanding of these processes to be able to analyse key elements, such as the PGE and gold, in the parts per billion to parts per trillion range. Platinum group elements and Au were determined by radiochemical neutron activation analysis using a modified NiS fire-assay preconcentration technique, adapted from procedures first used by Robert, R.V. D. and van Wyk, E. (1975) . Detection limits are generally 0.005-0.01 ppb (Au and Ir), 0.1-0.2 ppb (Pd and Pt), and 0.1-0.5 ppb for Ru. 9 refs.

  16. The platinum group elements and gold: analysis by radiochemical and instrumental neutron activation analysis and relevance to geological exploration and related problems

    International Nuclear Information System (INIS)

    Reeves, S.; Plimer, I. R.

    1996-01-01

    This paper presents an overview of research conducted with the support of the Australian Institute of Nuclear Science and Engineering, at the University of Melbourne, School of Earth Sciences, Radiochemical Neutron Activation Laboratory. The primary objective of this research is to realize the high potential of the platinum group elements (PGE) and gold to the solution of petrogenetic problems, the study of magma generation and magmatic processes in mafic/ultramafic rock suites, as tracers in hydrothermal ore formation. The PGEs (Os, Ru, Ir, Pt, Pd and Rh) are among the least abundant of all elements on earth with unique properties such as high melting points, high electrical and thermal conductivity, high density, strength and toughness as alloys. They exhibit both siderophile and chalcophile characteristics and are valuable tools in providing information about magmatic processes, in particular S-saturation, as well as crystal fractionation trends. Two distinct groups of PGEs are discerned; the IPGEs (Ru, Os, Ir) and the PPGEs (Pt, Pd, Rh, Au) on the basis of their behaviour during fractionation processes. Using chondrite normalized PGE patterns it is possible to distinguish between sulphides that segregated from primitive magmas, such as komatiites, and sulphides which segregated from more fractionated magmas, such as tholeiites. It is critical to the understanding of these processes to be able to analyse key elements, such as the PGE and gold, in the parts per billion to parts per trillion range. Platinum group elements and Au were determined by radiochemical neutron activation analysis using a modified NiS fire-assay preconcentration technique, adapted from procedures first used by Robert, R.V. D. and van Wyk, E. (1975) . Detection limits are generally 0.005-0.01 ppb (Au and Ir), 0.1-0.2 ppb (Pd and Pt), and 0.1-0.5 ppb for Ru. 9 refs

  17. The platinum group elements and gold: analysis by radiochemical and instrumental neutron activation analysis and relevance to geological exploration and related problems

    Energy Technology Data Exchange (ETDEWEB)

    Reeves, S; Plimer, I R [Melbourne Univ., Parkville, VIC (Australia). School of Physics

    1997-12-31

    This paper presents an overview of research conducted with the support of the Australian Institute of Nuclear Science and Engineering, at the University of Melbourne, School of Earth Sciences, Radiochemical Neutron Activation Laboratory. The primary objective of this research is to realize the high potential of the platinum group elements (PGE) and gold to the solution of petrogenetic problems, the study of magma generation and magmatic processes in mafic/ultramafic rock suites, as tracers in hydrothermal ore formation. The PGEs (Os, Ru, Ir, Pt, Pd and Rh) are among the least abundant of all elements on earth with unique properties such as high melting points, high electrical and thermal conductivity, high density, strength and toughness as alloys. They exhibit both siderophile and chalcophile characteristics and are valuable tools in providing information about magmatic processes, in particular S-saturation, as well as crystal fractionation trends. Two distinct groups of PGEs are discerned; the IPGEs (Ru, Os, Ir) and the PPGEs (Pt, Pd, Rh, Au) on the basis of their behaviour during fractionation processes. Using chondrite normalized PGE patterns it is possible to distinguish between sulphides that segregated from primitive magmas, such as komatiites, and sulphides which segregated from more fractionated magmas, such as tholeiites. It is critical to the understanding of these processes to be able to analyse key elements, such as the PGE and gold, in the parts per billion to parts per trillion range. Platinum group elements and Au were determined by radiochemical neutron activation analysis using a modified NiS fire-assay preconcentration technique, adapted from procedures first used by Robert, R.V. D. and van Wyk, E. (1975) . Detection limits are generally 0.005-0.01 ppb (Au and Ir), 0.1-0.2 ppb (Pd and Pt), and 0.1-0.5 ppb for Ru. 9 refs.

  18. A 90-day dietary toxicity study of genetically modified rice T1C-1 expressing Cry1C protein in Sprague Dawley rats.

    Directory of Open Access Journals (Sweden)

    Xueming Tang

    Full Text Available In a 90-day study, Sprague Dawley rats were fed transgenic T1C-1 rice expressing Cry1C protein and were compared with rats fed non-transgenic parental rice Minghui 63 and rats fed a basal diet. No adverse effects on animal behavior or weight gain were observed during the study. Blood samples were collected and analyzed, and standard hematological and biochemical parameters were compared. A few of these parameters were found to be significantly different, but were within the normal reference intervals for rats of this breed and age, and were thus not considered to be treatment-related. Following sacrifice, a large number of organs were weighed, and macroscopic and histopathological examinations were performed with no changes reported. The aim of this study was to use a known animal model to determine the safety of the genetically modified (GM rice T1C-1. The results showed no adverse or toxic effects due to T1C-1 rice when tested in this 90-day study.

  19. A 90-day dietary toxicity study of genetically modified rice T1C-1 expressing Cry1C protein in Sprague Dawley rats.

    Science.gov (United States)

    Tang, Xueming; Han, Fangting; Zhao, Kai; Xu, Yan; Wu, Xiao; Wang, Jinbin; Jiang, Lingxi; Shi, Wei

    2012-01-01

    In a 90-day study, Sprague Dawley rats were fed transgenic T1C-1 rice expressing Cry1C protein and were compared with rats fed non-transgenic parental rice Minghui 63 and rats fed a basal diet. No adverse effects on animal behavior or weight gain were observed during the study. Blood samples were collected and analyzed, and standard hematological and biochemical parameters were compared. A few of these parameters were found to be significantly different, but were within the normal reference intervals for rats of this breed and age, and were thus not considered to be treatment-related. Following sacrifice, a large number of organs were weighed, and macroscopic and histopathological examinations were performed with no changes reported. The aim of this study was to use a known animal model to determine the safety of the genetically modified (GM) rice T1C-1. The results showed no adverse or toxic effects due to T1C-1 rice when tested in this 90-day study.

  20. Are serum gamma-glutamyl transferase, high-sensitivity C-reactive protein, and ischaemia-modified albumin useful in diagnosing PCOS?

    Science.gov (United States)

    Ozturk, Mustafa; Keskin, Ugur; Ozturk, Ozlem; Ulubay, Mustafa; Alanbay, İbrahim; Aydin, Aytekin; Yenen, Müfit Cemal

    2016-10-01

    We assessed the serum levels of gamma-glutamyl transferase (GGT), high-sensitivity C-reactive protein (hsCRP) and ischaemia-modified albumin (IMA) in patients with polycystic ovary syndrome (PCOS). Fifty-three patients with PCOS were included in our study along with 40 women with no PCOS as the control group. The patients were divided according to their body mass index (BMI). GGT levels were significantly higher in the women with PCOS than the women in the control group (p PCOS women who were normoweight and overweight than the normoweight and overweight women in the control group (p PCOS and the controls or between the normoweight and overweight subgroups. GGT may be associated with the diagnosis of PCOS when the threshold is set at >15.5 U/L. With the application of this threshold, raised GGT levels had 83% sensitivity (95% CI 0.70-0.90) and 67.5% specificity (95% CI 0.52-0.79), for the diagnosis of PCOS. In our study, GGT levels were elevated in the PCOS patients independent of BMI and could thus be an important marker of PCOS.

  1. Clinical diagnostic value of determination of serum lschemia modified albumin, homocysteine and high-sensitivity C-reactive protein levels in patients with acute coronary syndrome

    International Nuclear Information System (INIS)

    Lu Weiqun; Jiang Donglin; Lu Weigu

    2009-01-01

    Objective: To study the clinical diagnostic value of determination of serum ischemia modified albumin (IMA), homocysteine (Hcy) and high-sensitivity C-reactive protein (hs-CRP) levels in patients with acute coronary syndrome (ACS). Methods: Serum IMA (with albumin-cobalt binding assay), Hcy (with ELISA) and hs-CRP (with RIA) levels were determined in 73 patients with ACS within 3 hours after onset of chest pain and 40 controls. Results: The sensitivity of IMA was 87.67%, the specificity was 100%. The area under curve of ROC was 0.985 (95% confidence interval 0.969 ∼ 1.001). All the figures were higher than those of Hcy and hs-CRP. Combined determination of IMA, Hcy and hs-CRP, would increass the sensitivity to 97.26%. Conclusion: IMA is one of the earliest sensitive indicators for clinical diagnosis of early myocardial ischemia in patients with ACS. Combined determination of IMA, Hcy and hs-CRP would be even more sensitive. (authors)

  2. Determination of chromium, cobalt and nickel in tissue samples by radiochemical activation analysis

    International Nuclear Information System (INIS)

    Reisell, A.; Lakomaa, E.L.

    1983-03-01

    A radiochemical neutron activation analysis method for the determination of chromium, cobalt and nickel in tissue samples. A radiochemical neutron activation analysis method for the determination of chromium, cobalt and nickel has been developed. The destruction device used consisted of a combined wet-ashing-distillation and ion-exchange system. Six samples could be treated at the same time. The samples were wet-ashed with H*L2SO*L4-H*L2O*L2 mixture. Volatile elements were distilled as bromide compounds with HBr*H-. The distillation residue in 8M HCl was passed through hydrated antimony pentoxide (HAP) in order to remove disturbing *H2*H4Na-activity and through a Dowex 2 x 8 column so as to retain *H6*H0Co (formed from *H5*H8Ni). Chromium was elutriated from the column and precipitated as Cr(OH)*L3 for the removal of disturbing *H3*H2P-activity. The standards and samples were treated in a similar manner each so that the yield determination is not necessarily needed. The yields by tracer experiments were (43 +- 5) % for Cr, (93 +- 4) % for Co and (88 +- 14) % for Ni. The precision and accuracy of the method were studied by using reference materials of the National Bureau of Standards (NBS) and the International Atomic Energy Agency (IAEA)

  3. Growth of a manganese nodule from Peru Basin: a radiochemical anatomy

    International Nuclear Information System (INIS)

    Reyss, J.L.; Lemaitre, N.; Marchig, V.; Southon, J.R.; Nelson, D.E.; Vogel, J.S.

    1985-01-01

    Attempts have been made to study the entire growth history of a manganese nodule from the northern part of Peru Basin in the Pacific using radiochemical profiles of 230 Th/ 232 Th, 227 Th/ 230 Th and 10 Be/ 9 Be. Combined with the observations on Fe-Mn contents and textural variation, the radiochemical data indicate that the nodule grew more or less concentrically throughout most of its existence since it formed 1.5 my ago, receiving Mn from both bottom water and pore water. This condition appeared to have changed about 180 ky ago when the growth became asymmetric in that the top and bottom sides became fixed in their relative positions on the sea floor. Since then, the bottom side accreted with a fast rate of close to 200 mm/my, apparently fueled by the supply of diagenetically remobilized Mn in pore water from the sediment substrate. In the meantime, the topside accumulated at about 6 mm/my, a value which is in the normal range for deep-sea nodules having their Mn supplied from the hydrogenous source. (author)

  4. Cyclotron production, radiochemical separation and quality control of platinum radiotracers for toxicological studies

    International Nuclear Information System (INIS)

    Bonardi, M.; Birattari, C.; Groppi, F.; Arginelli, D.; Gini, L.; Gallorini, M.

    1998-01-01

    The increasing concentration of Pt, Pd and Rh in the environment is mainly due to the release of these elements from the catalytic converters of the motorvehicles. This situation makes it necessary to carry out metallotoxicological experiments on both cell cultures and laboratory animals, in order to assess their impact on living organisms after a Long Term and Low Level Exposure (LLE). Both nuclear reactions nat Ir(p,xn) and nat Os(α,xn) were investigated in the energy range up to 45 MeV for protons and 38 MeV for alpha-particles, in order to optimize the irradiation parameters for the production of 188,189,191 Pt. Several sets of thin- and thick-target excitation functions were determined experimentally by cyclotron irradiation at both Milano and Ispra cyclotrons. This paper reports the irradiation parameters studied and adopted and two radiochemical procedures for the separation of radio-Pt from an Os target, as well as from ruthenium, iridium and gold impurities. These procedures were used to obtain very high specific activity Pt radionuclides in No Carrier Added (NCA) form. Radionuclidic, radiochemical and chemical purity measurements were carried out by the use of several techniques like γ-spectrometry, ion-exchange radio-chromatography, atomic absorption spectrometry and neutron activation analysis. (author)

  5. RAPID AUTOMATED RADIOCHEMICAL ANALYZER FOR DETERMINATION OF TARGETED RADIONUCLIDES IN NUCLEAR PROCESS STREAMS

    International Nuclear Information System (INIS)

    O'Hara, Matthew J.; Durst, Philip C.; Grate, Jay W.; Egorov, Oleg; Devol, Timothy A.

    2008-01-01

    Some industrial process-scale plants require the monitoring of specific radionuclides as an indication of the composition of their feed streams or as indicators of plant performance. In this process environment, radiochemical measurements must be fast, accurate, and reliable. Manual sampling, sample preparation, and analysis of process fluids are highly precise and accurate, but tend to be expensive and slow. Scientists at Pacific Northwest National Laboratory (PNNL) have assembled and characterized a fully automated prototype Process Monitor instrument which was originally designed to rapidly measure Tc-99 in the effluent streams of the Waste Treatment Plant at Hanford, WA. The system is capable of a variety of tasks: extraction of a precise volume of sample, sample digestion/analyte redox adjustment, column-based chemical separations, flow-through radiochemical detection and data analysis/reporting. The system is compact, its components are fluidically inter-linked, and analytical results can be immediately calculated and electronically reported. It is capable of performing a complete analytical cycle in less than 15 minutes. The system is highly modular and can be adapted to a variety of sample types and analytical requirements. It exemplifies how automation could be integrated into reprocessing facilities to support international nuclear safeguards needs

  6. Validation of the FDG (18F) radiochemical purity assay by thin layer chromatography

    International Nuclear Information System (INIS)

    Leao, R.L.C.; Oliveira, M.L.; Nascimento, J.E.; Nascimento, N.C.E.S.

    2013-01-01

    All methodologies utilized in radiopharmaceutical industry should be validated in order to prove that they meet the requirements of analytical applications, ensuring the reliability of the results. At a radiopharmaceutical industry there is one challenge aspect: sometimes it is not possible use a stable standard to perform the validation analysis. In order to overcome this difficulty, the objective of this study was to suggest a validation protocol for these methodologies, based on the recommendations of RE n° 899/Agencia Nacional de Vigilancia Sanitaria (ANVISA), and prove its efficiency, performing the radiochemical purity validation test of FDG (18F), by TLC. To obtain the calibration curve, we suggested that the theoretical activity values should be determined using a dose calibrator, simultaneously of each analysis performed by TLC, for 5 hours. The method was linear (R 2 of 0.996), precise (CV% <5%) and accurate (96.85% < accuracy < 102.56%). In relation to the robustness test, our experiments evaluated the influence of the distance travelled by mobile phase, variations at mobile phase concentration and type of chromatographic plate (silica gel on glass or aluminium plates). The detection and quantification limits were determined (321.9 and 1065.6 kBq, respectively). As expected, this methodology was nonspecific, showing a slight spot corresponding to the FDM. The proposed protocol was efficient and the methodology tested was effective to determine the radiochemical purity of FDG (18F), in accordance to the limits recommended by ANVISA. (author)

  7. A direct reading on-line flowrate meter for use in radiochemical plant

    International Nuclear Information System (INIS)

    Shah, B.V.; Kaimal, C.K.R.; Siddiqui, I.A.; Kumar, S.V.

    1987-01-01

    A device for measurement and remote direct reading display of the flowrates of streams in a radiochemical plant is described. The device is interposed in the measured stream and consists of a syphon pot with a specially developed attachment on the discharge line. Differential pressure switches are used to trigger a timer device at set levels in the pot and the time required for filling the pot during each cycle is measured and is used to compute and display the flowrate. The device is accurate and reliable and is simple to fabricate and install. It is maintenance-free since it has no moving parts. It is also suggested that a manometer with conductive contacts could be used in place of the d.p. switches. The background and various stages of development of the device are described. The operating data is tabulated and parameters required for plant applications are indicated in detail. A simple method to detect and correct for errors due to drift in d.p. switch setting is also outlined. Sketches of typical syphon pot, the schematic of the apparatus and suggested layout for application in radiochemical plant are also included. (author). 11 figures, 6 tables

  8. The use of cuprous iodide as a precipitation matrix in the radiochemical determination of 131I in milk

    International Nuclear Information System (INIS)

    McCurdy, D.E.; Mellor, R.A.; Lambdin, R.W.; McLain, M.E. Jr.

    1980-01-01

    As a result of the implementation of the As Low As is Reasonably Achievable philosophy to the nuclear power industry, recent U.S. Nuclear Regulatory Commission requirements have prompted high sensitivity radiochemical analysis for the measurement of 131 I in milk. The most recognized and commonly employed technique incorporates costly palladium iodide as the final precipitate in the radiochemical purification of the iodine chemical species. The procedure presented in this paper outlines the many advantages of using cuprous iodide as the final precipitate. These include lower cost per analysis, consistent recoveries, better precipitate matrix and good self absorption characteristics. Typical lower limit of detection values and operating characteristics obtained for high sensitivity β-γ analysis as well as gas proportional counting and a comparison of radiochemical and Ge(Li) spectrometric results for environmental samples collected during a recent Chinese weapons fallout incident are presented. (author)

  9. Trace analysis measurements in high-purity aluminium by means of radiochemical neutron and proton activation analysis

    International Nuclear Information System (INIS)

    Egger, K.P.

    1987-01-01

    The aim of the study consisted in the development of efficient radiochemical composite processes and activation methods for the multi-element determination of traces within the lower ng range in high-purity aluminium. More than 50 elements were determined with the help of activation with reactor neutrons; the selective separation of matrix activity (adsorption with hydrated antimony pentoxide) led to a noticeable improvement of detectability, as compared with instrumental neutron activation analysis. Further improvements were achieved with the help of radiochemical group separations in ion exchangers or with the help of the selective separation of the pure beta-emitting elements. Over 20 elements up to high atomic numbers were determined by means of activating 13 MeV protons and 23 Me protons. In this connection, improvements of the detection limit by as a factor of 10 were achieved with radiochemical separation techniques, as compared with pure instrumental proton activation analysis. (RB) [de

  10. Modified cyanobacteria

    Science.gov (United States)

    Vermaas, Willem F J.

    2014-06-17

    Disclosed is a modified photoautotrophic bacterium comprising genes of interest that are modified in terms of their expression and/or coding region sequence, wherein modification of the genes of interest increases production of a desired product in the bacterium relative to the amount of the desired product production in a photoautotrophic bacterium that is not modified with respect to the genes of interest.

  11. HPLC-ICP-MS compared with radiochemical detection for metabolite profiling of H-3-bromohexine in rat urine and faeces

    DEFF Research Database (Denmark)

    Jensen, B.P.; Gammelgaard, B.; Hansen, S.H.

    2005-01-01

    H-3-Bromohexine was dosed to rats as a model compound to allow comparison of HPLC-ICP-MS detection on bromine to radiochemical detection in an in vivo drug metabolism study. Metabolite profiles were obtained in urine and faeces extracts. No influence of the methanol gradient on the bromine response...... was observed in the range of 18 - 75% methanol. The sensitivity obtained by HPLC- ICP-MS was almost two orders of magnitude better than on-line H-3 radiochemical detection. For ICP- MS, the limit of detection was calculated to be 69 nM Br ( injection volume 100 mu l), corresponding to an absolute limit...

  12. Determination of gold and platinum in biological materials by radiochemical neutron activation analysis using electrolytic separation of gold

    International Nuclear Information System (INIS)

    Reitz, B.; Heydorn, K.

    1993-01-01

    A new method is presented for the determination of Au and Pt in biological materials based on neutron activation analysis with radiochemical separation of gold. Separation of gold by electrolytic deposition on a niobium cathode ascertains thee highest radiochemical purity without any interference from calcium or other major elements. With 199 Au as indicator for platinum the gold content of the sample not only strongly affects the limit of detection, but also causes interference by double neutron capture. Replicate analyses of BCR Certified Reference Materials No. 184, 185 and 186 were carried out. (author) 18 refs.; 3 figs.; 2 tabs

  13. Function of the cytoplasmic tail of human calcitonin receptor-like receptor in complex with receptor activity-modifying protein 2

    Energy Technology Data Exchange (ETDEWEB)

    Kuwasako, Kenji, E-mail: kuwasako@fc.miyazaki-u.ac.jp [Frontier Science Research Center, University of Miyazaki, 5200 Kihara, Kiyotake, Miyazaki 889-1692 (Japan); Kitamura, Kazuo; Nagata, Sayaka; Hikosaka, Tomomi [Division of Circulation and Body Fluid Regulation, Faculty of Medicine, University of Miyazaki, 5200 Kihara, Kiyotake, Miyazaki 889-1692 (Japan); Kato, Johji [Frontier Science Research Center, University of Miyazaki, 5200 Kihara, Kiyotake, Miyazaki 889-1692 (Japan)

    2010-02-12

    Receptor activity-modifying protein 2 (RAMP2) enables calcitonin receptor-like receptor (CRLR) to form an adrenomedullin (AM)-specific receptor. Here we investigated the function of the cytoplasmic C-terminal tail (C-tail) of human (h)CRLR by co-transfecting its C-terminal mutants into HEK-293 cells stably expressing hRAMP2. Deleting the C-tail from CRLR disrupted AM-evoked cAMP production or receptor internalization, but did not affect [{sup 125}I]AM binding. We found that CRLR residues 428-439 are required for AM-evoked cAMP production, though deleting this region had little effect on receptor internalization. Moreover, pretreatment with pertussis toxin (100 ng/mL) led to significant increases in AM-induced cAMP production via wild-type CRLR/RAMP2 complexes. This effect was canceled by deleting CRLR residues 454-457, suggesting Gi couples to this region. Flow cytometric analysis revealed that CRLR truncation mutants lacking residues in the Ser/Thr-rich region extending from Ser{sup 449} to Ser{sup 467} were unable to undergo AM-induced receptor internalization and, in contrast to the effect on wild-type CRLR, overexpression of GPCR kinases-2, -3 and -4 failed to promote internalization of CRLR mutants lacking residues 449-467. Thus, the hCRLR C-tail is crucial for AM-evoked cAMP production and internalization of the CRLR/RAMP2, while the receptor internalization is dependent on the aforementioned GPCR kinases, but not Gs coupling.

  14. A radio-high-performance liquid chromatography dual-flow cell gamma-detection system for on-line radiochemical purity and labeling efficiency determination

    DEFF Research Database (Denmark)

    Lindegren, S; Jensen, H; Jacobsson, L

    2014-01-01

    In this study, a method of determining radiochemical yield and radiochemical purity using radio-HPLC detection employing a dual-flow-cell system is evaluated. The dual-flow cell, consisting of a reference cell and an analytical cell, was constructed from two PEEK capillary coils to fit into the w...

  15. Metrological aspects of radiochemical methods for determining activity of gamma-emitting nuclides

    International Nuclear Information System (INIS)

    Shcherbakov, B.Y.

    1986-01-01

    The author considers the problem of metrological compatibility of the two stages in the radiochemical method of determining the activity of a gamma-emitting nuclide: chemical isolation of the nuclide and radiometric measurement of its activity. The authors show that preparation of the specimen in liquid form provides for important advantages compared with the traditional application of the solid residue onto a flat substate. The work here is of interest for analytical chemists who are involved with determination of the activity of gamma emitting nuclides such as Ru 103, Rh 106, Sn 113, Cs 134, Cs 137, La 140, Ce 141, Ce 144, Hg 203, Na 24, Mn 54, Fe 59, Co 60, Zn 65, Zr 95, and Nb 95, for example, in waste water or in emissions to the atmosphere, with the goal of protecting the environment

  16. Hanford Environmental Restoration data validation process for chemical and radiochemical analyses

    International Nuclear Information System (INIS)

    Adams, M.R.; Bechtold, R.A.; Clark, D.E.; Angelos, K.M.; Winter, S.M.

    1993-10-01

    Detailed procedures for validation of chemical and radiochemical data are used to assure consistent application of validation principles and support a uniform database of quality environmental data. During application of these procedures, it was determined that laboratory data packages were frequently missing certain types of documentation causing subsequent delays in meeting critical milestones in the completion of validation activities. A quality improvement team was assembled to address the problems caused by missing documentation and streamline the entire process. The result was the development of a separate data package verification procedure and revisions to the data validation procedures. This has resulted in a system whereby deficient data packages are immediately identified and corrected prior to validation and revised validation procedures which more closely match the common analytical reporting practices of laboratory service vendors

  17. The determination of the radiochemical purity of 99Tcm-DTPA-HSA injection

    International Nuclear Information System (INIS)

    Yin Guangrong; Zhang Yan

    1996-01-01

    A simple, rapid analytical method of radiochemical purity for the 99 Tc m -DTPA-HSA injection is established. 99 Tc m (VII), 99 Tc m (IV) and labelled compound 99 Tc m -DTPA-HSA are separated by two systems of paper chromatography on Whatman No.1. The solvent of the system A is 85% methanol and that of the system B is water-95% alcohol-ammonium hydroxide (volume ratio is 5:2:1). 90 Tc m (IV) and labelled compound are located at the origin, while R f of 99 Tc m (VII) is about 0.4-0.5 in the system A. R f of 99 Tc m (VII) and labelled compound are about 0.7-0.8, while 99 Tc m (IV) is located at the origin in the system B

  18. Limits of the radiochemical and geochemical method for the test of nucleon stability

    Energy Technology Data Exchange (ETDEWEB)

    Bergamasco, L [CNR, Laboratorio di Cosmo-Geofisica, Turin, Italy; Texas A and M University, College Station, Tex.); Cini, G [CNR, Laboratorio di Cosmo-Geofisica; Torino, Universita, Turin, Italy)

    1978-07-01

    An estimate is obtained for the limiting nucleon lifetime that can be determined by the radiochemical method of Steinberg and Evans (1977), which counts the Ar-37 production from reactions due to nucleon decay in a large sample of KC2H3O2. To estimate the background rate, processes due to the atmospheric muon component are considered. It is found that the asymptotic limit on the maximum value of the nucleon lifetime is 2 x 10/sup 30/years. This value cannot be increased by going to larger depths or by using a larger sample, but is inherent in the method itself. To improve the limit one would have to resort to methods based on multiparticle decay modes.

  19. Radiochemical separation and effective dose estimation due to ingestion of 90Sr

    International Nuclear Information System (INIS)

    Ilic, Z.; Vidic, A.; Deljkic, D.; Sirko, D.; Zovko, E.; Samek, D.

    2009-01-01

    Since 2007. Institute for Public Health of Federation of Bosnia and Herzegovina-Radiation Protection Centre, within the framework of monitoring of radioactivity of environment carried out measurement of specific activity of 90 Sr content in selected food and water samples. The paper described the methods of measurement and radiochemical separation. Presented results, as average values of specific activity of 90 Sr, were used for estimation of effective dose due to ingestion of 90 Sr for 2007. and 2008. Estimated effective dose for 2007. due to ingestion of 90 Sr for adults was 1,36 μSv and 2,03 μSv for children (10 year old), and for 2008. 0,67 μSv (adults) and 1,01 μSv (children 10 year old). Estimated effective doses for 2007. and 2008. are varied because of different average specific activity radionuclide 90 Sr in selected samples of food, their number, species and origin. (author) [sr

  20. Determination of radiochemical yields of 186Re-labelled complexes using thin layer chromatography

    International Nuclear Information System (INIS)

    Konirova, R.; Kohlickova, M.; Jedinakova-Krizova, V.

    1999-01-01

    The reaction conditions for synthesis of three rhenium complexes 186 Re-methylendiphosphonate (MDP), 186 Re-hydroxyethylidendiphosphonate (HEDP) and 186 Re-citrate have been investigated. Radiochemical yield of complexation has been determined by thin layer chromatography and paper chromatography. The rhenium complexation with corresponding ligand is dependent on pH values of reaction mixture, concentration of studied ligand (MDP, HEDP and sodium citrate) and concentration of reducing agent. Stannous chloride with ascorbic acid (as antioxidant) was used for reduction of perrhenate. The labeling yield of 186 Re-MDP was about 90 %, of 186 Re-HEDP more than 80 % and more than 75 % for 186 Re-citrate under optimum conditions. Besides, the possibility of application of porphyrins as organic ligands for complexation with rhenium isotopes is examined. (authors)

  1. Development of radiochemical separation method for determination of toxic elements in biological samples

    International Nuclear Information System (INIS)

    Maihara, V.A.; Vasconcellos, M.B.A.; Favaro, D.I.T.; Armelin, M.J.A.

    1990-01-01

    In order to determine Hg, Sb, As and Se in biological materials by neutron activation analysis, a radiochemical separation was developed. The chemical separation procedure used was based on the digestion of the irradiated sample in a mixture of H NO 3 and H 2 SO 4 in a teflon bomb, at 130 0 C for 1 to 4 hours. After the dissolution of organic matter, Hg and Sb were retained by a Dowex 2-X8 resin column in 6 M HCl. The effluent was passed through a TDO, tin dioxide column which retains As and Se in 3 M HCl medium. Radioactive tracers of these elements were used to determine the yields of the separation process. Certified reference materials were analyzed to check the precision and accuracy of the method. (author)

  2. Development of a radiochemical separation method for toxic elements determination from biologic samples

    International Nuclear Information System (INIS)

    Malhara, V.A.; Vasconcellos, M.B.A.; Favaro, D.I.T.; Armelin, M.J.A.

    1990-01-01

    In order to determine Hg, Sb, As and Se in biological materials by neutron activation analysis, a radiochemical separation was developed. The chemical separation procedure used was based on the digestion of the irradiated sample in a mixture of HNO 3 and H 2 SO 4 in a teflon bomb, at 130 0 C for 1 to 4 hours. After the dissolution of organic matter, Hg and Sb were retained by a Dowex 2-XB resin column in 6M HCI. The efluent was passed through a TDO, tin dioxide column which retains As and Se in 3M HCI medium. Radioactive tracers of these elements were used to determine the yields of the separation process. Certified reference materials were analyzed to check the precision and accuracy of the method. (author)

  3. Investigation of the possibility of using hydrogranulation in reprocessing radioactive wastes of radiochemical production facilities

    Energy Technology Data Exchange (ETDEWEB)

    Revyakin, V.; Borisov, L.M. [All Russian Scientific and Research Institute of Non-Organic Materials, Moscow (Russian Federation)

    1996-05-01

    Radio-chemical production facilities are constantly accumulating liquid radioactive wastes (still residues as the result of evaporation of extraction and adsorption solutions etc.) which are a complex multicomponent mixtures. The wastes are frequently stored for extended periods of time while awaiting disposition and in some cases, and this is much worse, they are released into the environment. In this report, I would like to draw your attention to some results we have obtained from investigations aimed at simplifying handing of such wastes by the precipitation of hard to dissolve metal hydroxides, the flocculation of the above into granules with the help of surface-active agents (in this case a polyacrylamide - PAA), quickly precipitated and easily filtered. The precipitate may be quickly dried and calcinated, if necessary, and transformed into a dense oxide sinter. In other words it may be transformed into a material convenient for storage or burial.

  4. Radiochemical determination of Inertial Confinement Fusion capsule compression at the National Ignition Facility

    Energy Technology Data Exchange (ETDEWEB)

    Shaughnessy, D. A., E-mail: shaughnessy2@llnl.gov; Moody, K. J.; Gharibyan, N.; Grant, P. M.; Gostic, J. M.; Torretto, P. C.; Wooddy, P. T.; Bandong, B. B.; Cerjan, C. J.; Hagmann, C. A.; Caggiano, J. A.; Yeamans, C. B.; Bernstein, L. A.; Schneider, D. H. G.; Henry, E. A.; Fortner, R. J. [Lawrence Livermore National Laboratory, 7000 East Avenue, Livermore, California 94551 (United States); Despotopulos, J. D. [Lawrence Livermore National Laboratory, 7000 East Avenue, Livermore, California 94551 (United States); Radiochemistry Program, University of Nevada Las Vegas, Las Vegas, Nevada 89154 (United States)

    2014-06-15

    We describe a radiochemical measurement of the ratio of isotope concentrations produced in a gold hohlraum surrounding an Inertial Confinement Fusion capsule at the National Ignition Facility (NIF). We relate the ratio of the concentrations of (n,γ) and (n,2n) products in the gold hohlraum matrix to the down-scatter of neutrons in the compressed fuel and, consequently, to the fuel's areal density. The observed ratio of the concentrations of {sup 198m+g}Au and {sup 196g}Au is a performance signature of ablator areal density and the fuel assembly confinement time. We identify the measurement of nuclear cross sections of astrophysical importance as a potential application of the neutrons generated at the NIF.

  5. Radiochemical purity and in vitro stability of Tc-99m radiopharmaceuticals

    International Nuclear Information System (INIS)

    Vucina, J.

    2001-01-01

    The increased contents of long lived 99 Tc, oxygen and cupric ions could affect the labeling yield of eight radiopharmaceuticals. Oxygen and in leaser extend copper were found to affect the radiochemical purity of the preparations. In vitro stability of radiopharmaceuticals, examined on 99m Tc(Sn)-pyrosphoshate solutions, was extended when ascorbic acid was added as the chemical stabilizer. The quantity of 5x10 -7 mol/dm 3 of ascorbic acid was found to be sufficient to keep the content of 99m Tc-pertechnetate below 1 % six hours after labeling even in the cases when 99m Tc was present in high radioactive concentrations (740-814 GBq/dm 3 ). The results led to the development of the kits in which ascorbic or gentisic acid are the standards component in the kit composition (author)

  6. Preparation, radiochemical analysis and biodistribution of 99mTc-dihydrobis(1-pyrazolyl)borate

    International Nuclear Information System (INIS)

    Owunwanne, A.; Abdel-Dayem, H.; Yacoub, T.

    1987-01-01

    Optimum preparation of 99m Tc-dihydrobis(1-pyrazolyl)borate ( 99m Tc-HBPz 2 ) was done by mixing 1.4 mg/ml HBPz 2 and 1.0 mg/ml of stanous PYP. Radiochemical analysis of the preparation using paper chromatography (PC), thin layer chromatography (TLC) and high performance liquid chromatography (HPLC) indicated a stable product with one major component. The labelling efficiency was approximately 90%. Animal biodistribution studies performed in mice showed that most of the injected radioactivity was confined to the liver, kidney, lungs, intestine and heart. The heart to blood ratio was small but persisted up to 3 hrs. after the injection. (orig.) [de

  7. Radiochemical analyses of surface water from U.S. Geological Survey hydrologic bench-mark stations

    Science.gov (United States)

    Janzer, V.J.; Saindon, L.G.

    1972-01-01

    The U.S. Geological Survey's program for collecting and analyzing surface-water samples for radiochemical constituents at hydrologic bench-mark stations is described. Analytical methods used during the study are described briefly and data obtained from 55 of the network stations in the United States during the period from 1967 to 1971 are given in tabular form.Concentration values are reported for dissolved uranium, radium, gross alpha and gross beta radioactivity. Values are also given for suspended gross alpha radioactivity in terms of natural uranium. Suspended gross beta radioactivity is expressed both as the equilibrium mixture of strontium-90/yttrium-90 and as cesium-137.Other physical parameters reported which describe the samples include the concentrations of dissolved and suspended solids, the water temperature and stream discharge at the time of the sample collection.

  8. Radiochemical method for evaluating the effect of antibiotics on Escherichia coli biofilms

    International Nuclear Information System (INIS)

    Dix, B.A.; Cohen, P.S.; Laux, D.C.; Cleeland, R.

    1988-01-01

    A simple radiochemical method for evaluating the action of antibiotics on Escherichia coli cells in biofilms is reported. After growth, biofilms of E. coli ATCC 25922 on disks of urinary catheter material were suspended in fresh medium containing or lacking an antibiotic, incubated for 4 h at 37 degrees C, and pulse-labeled with [ 3 H]leucine for 5 min. Radioactivity in trichloracetic acid-precipitable material in the biofilm and in the surrounding medium (planktonic E. coli) was then measured. Antibiotic-induced inhibition of incorporation of [ 3 H]leucine into the cells in the biofilm was far less pronounced than incorporation into planktonic cells and, furthermore, correlated well with loss in viable counts. The method is simple, inexpensive, and extremely timesaving

  9. Radiochemical neutron activation analysis for trace elements evaluation of human milk

    International Nuclear Information System (INIS)

    Gill, K.P.; Zaidi, J.H.; Ahmad, S.

    2003-01-01

    The principal objective pursued in this study is to establish the base-line data on the status of elemental composition in human milk from Pakistani subjects of Rawalpindi/Islamabad area. Radiochemical neutron activation analysis (RNAA) methodology was developed and successfully employed to determine the concentration of 18 minor and trace elements (essential, toxic and nonessential) in human milk. This methodology has significantly improved the detection limits of most of these elements due to suppression of Compton background. The data provide the base-line values of these elements in human milk of low- and medium-income group subjects of the region. The results obtained show good compatibility with the data reported by the WHO on elemental composition of human milk from different geological regions. (orig.)

  10. Standard test methods for chemical, mass spectrometric, spectrochemical, nuclear, and radiochemical analysis of uranium hexafluoride

    CERN Document Server

    American Society for Testing and Materials. Philadelphia

    2011-01-01

    1.1 These test methods cover procedures for subsampling and for chemical, mass spectrometric, spectrochemical, nuclear, and radiochemical analysis of uranium hexafluoride UF6. Most of these test methods are in routine use to determine conformance to UF6 specifications in the Enrichment and Conversion Facilities. 1.2 The analytical procedures in this document appear in the following order: Note 1—Subcommittee C26.05 will confer with C26.02 concerning the renumbered section in Test Methods C761 to determine how concerns with renumbering these sections, as analytical methods are replaced with stand-alone analytical methods, are best addressed in subsequent publications. Sections Subsampling of Uranium Hexafluoride 7 - 10 Gravimetric Determination of Uranium 11 - 19 Titrimetric Determination of Uranium 20 Preparation of High-Purity U3O 8 21 Isotopic Analysis 22 Isotopic Analysis by Double-Standard Mass-Spectrometer Method 23 - 29 Determination of Hydrocarbons, Chlorocarbons, and Partially Substitut...

  11. The study of radiochemical separation methods on gaseous Fission product krypton-88

    International Nuclear Information System (INIS)

    Yang Zhihong; Zhang Shengdong; Yang Lei; Ding Youqian; Sun Hongqing; Ma Peng

    2012-01-01

    Half-life of krypton-88 is 2.84 hours, high fission yields and a relatively large gamma branching ratio is had. The gas is short-lived fission products in burnup measurements. Only New fission products can extract from extraction in gas of fissile irradiation target. But krypton-88 with krypton-85, krypton-87, xenon -135, and xenon-138 is coexisted together, thus radiochemical separation must quickly taken. selected the irradiation time is 1-2 hours and cooling time is best 2 hours for sample preparation, krypton and xenon were separated using activated carbon adsorption, the ratio of krypton and xenon were measured by gamma spectroscopy. Then according to the ratio of krypton-85 and xenon-125 count rate coefficient around separation were calculated yield of krypton and decontamination factor of xenon and the final the yield of krypton-85 is calculated. (authors)

  12. Continuous radiochemical analysis of fission products in a nuclear reactor water coolant

    International Nuclear Information System (INIS)

    Moskvin, L.N.; Zakharov, L.K.; Leont'ev, G.G.; Mel'nikov, V.A.; Orlenkov, I.S.; Slutskij, G.K.

    1975-01-01

    Method for continuous radiochemical analysis of I, Cs, Ba, Sr and Ce isotopes in a reactor water heat-transfer agent was developed. A continuous two-dimensional chromatographic process of complex mixtures separation of substances proved to be feasible on several parallel sorbent layers, which moved at constant velocities and separated by stationary intermediate collectors. Tests on model solutions containing I, Ce, Cs and Ba isotopes and on heat-carrier samples showed quantitative separation of elements. The results were indicative of a basic possibility of using multisorbent chromatographs for continuous control of multicomponent mixtures, particularly for control of radioactive fission product compositions in water heat-transfer agents in nuclear power plants. A diagram is shown for a two-dimensional chromatographic separation of a multicomponent mixture. Also shown is a flow chart of an installation for continuous control of iodine and cesium isotope activities

  13. Determination of rhenium in biological and environmental samples by radiochemical neutron activation analysis

    International Nuclear Information System (INIS)

    Kucera, J.; Mizera, J.; Randa, Z.; Byrne, A.R.; Lucanikova, M.

    2006-01-01

    Radiochemical neutron activation procedures using liquid-liquid extraction with tetraphenylarsonium chloride in chloroform from 1 M HCl and solid extraction with ALIQUAT 336 incorporated in a polyacrylonitrile binding matrix from 0.1 M HCl were developed for accurate determination of rhenium in biological and environmental samples at the sub-ng.g -1 level. Concentrations of Re in the range of 0.1 to 2.4 ng.g -1 were determined in several botanical reference materials (RM), while in a RM of road dust a value of approx. 10 ng.g -1 was found. Significantly elevated values of Re, up to 90 ng.g -1 , were found in seaweed (brown algae). Results for Re in the brown algae Fucus vesiculosus in which elevated 99 Tc values had previously been determined suggest possible competition between Re and Tc in the accumulation process. (author)

  14. A radiochemical assay for detection of leukotriene B4 production from isolated cells

    International Nuclear Information System (INIS)

    Aharony, D.; Dobson, P.; Krell, R.D.

    1984-01-01

    A radiochemical procedure for quantitating the effect of inhibitors of leukotriene B4 (LTB4) biosynthesis is described. Rat peritoneal cells were labeled with 3 H-arachidonic acid and stimulated with the calcium ionophore A23187. 3 H-LTB4 was isolated by processing on C18-Sep Pak cartridges followed by reverse-phase high pressure liquid chromatography (HPLC). The identity of this product, as well as other arachidonic acid metabolites, was verified by using silicic acid column chromatography followed by straight-phase HPLC and thin layer chromatography. Using this assay, LTB4 release by ionophore A23187 has been shown to be both time- and concentration-dependent. Indomethacin enhanced, while NDGA and ETYA inhibited, the A23187-induced production of LTB4. This procedure is both simple and direct and is capable of assessing the ability of novel compounds to alter LTB4 production

  15. A radiochemical procedure for the determination of Po-210 in environmental samples

    International Nuclear Information System (INIS)

    Godoy, J.M.; Schuettelkopf, H.

    1980-07-01

    A radiochemical procedure for the determination of Po-210 in environmental samples was developed. Soil, sediments, filter materials, plants, water and food samples can be analyzed for Po-210. Wet ashing is achieved with HNO 3 + H 2 O 2 or HCl + HNO 3 . To separate disturbing substances, a coprecipitation with Te is used for sample materials containing silica. Po-210 deposition from HCl solution on Ag platelets with other sample materials is possible directly. Deposited Po-210 is counted by α-spectrometry. For chemical yield determination Po-208 is added, yields range between 60% and 100%. A lower detection limit of about 0,002 pCi Po-210/sample is achievable. (orig./HP) [de

  16. Determination of individual rare earth elements in Vietnamese monazite by radiochemical neutron activation analysis

    International Nuclear Information System (INIS)

    Nguyen Van Suc; Nguyen Mong Sinh

    1993-01-01

    Radiochemical neutron activation analysis (RNAA) has been applied for determination of rare earth elements (REE) in Vietnamese monazite. The chemical separation procedure used is based on the chromatographic elution of rare earth groups, after the separation of 233 Pa(Th) in irradiated monazite samples by coprecipitation with MnO 2 , the rare earth elements were retained by Biorad AG1 x 8 resin column in 10% 15.4M HNO 3 -90% methanol solution. The elution of heavy rare earth (HREE) and middle rare earth (MREE) groups was carried out with 10% 1M HNO 3 - 90% methanol and 10% 0.05M HNO 3 -90% methanol solution, respectively; while the light rare earths (LREE) were eluted from the column by 0.1M HNO 3 solution. The accuracy of the method was checked by the analysis of granodiorite GSP-I and the rare earth values were in good agreement. (author) 7 refs.; 3 tabs

  17. Comparison study among methodologies of planar chromatography for radiochemical control of technetium-99m

    International Nuclear Information System (INIS)

    Monteiro, Elisiane de Godoy

    2012-01-01

    Radiopharmaceuticals are substances that have radioisotopes in their composition. About 95% of the procedures performed in nuclear medicine use radiopharmaceuticals with diagnostic purposes, and the Lyophilized Reagents (LR) labeled with Technetium-99m ( 99 mTc), obtained from 99 Mo/ 99 mTc generator, are the most one used. Quality Control represents the set of assays to be performed to assure that the product is adequate to its purpose. An important feature to be evaluated in 99m Tc radiopharmaceuticals is the radiochemical purity (% RqP) to quantify free pertechnetate ( 99 mTcO 4 - ) and technetium colloidal (99mTcO 2 ) mainly by paper chromatography (PC), thin layer (TLC) and High Performance Liquid Chromatography (HPLC). The objective of this work was to perform the comparison among the radiochemical control methodologies of LR labeled with 99m Tc, described in the United States Pharmacopoeia (USP) and European Pharmacopoeia (EP) and those used by IPEN. 99m TcO 4 - eluate and DISIDA, DMSA, DTPA, EC, ECD, GHA, MIBI, MDP, PIRO, SAH and Sn Coloidal LR were provided by IPEN-CNEN/SP. TLC-cellulose, TLC-SG.TLC-SG reverse phase, HPTLC-cellulose, HPTLC-SG (Merck) and ITLC-SG (Pall Corporation), W1MM, W3MM, W17M e W31ET (Whatman) chromatographic plates were used. The measurement of the radioactivity was done in a Perkin Elmer Cobra D-5002 gamma counter. LR were labeled to obtain 55,0 MBq mL 1 (1,5 mCi mL 1 ) of final radioactive concentration. The % 99m TcO 4 - , % 99m TcO 2 and % RqP were determined up to 4 hour labeling. From 11 LR, only EC and GHA have no radiochemical control methods in USP and EP. In USP and/or EP, DTPA, MDP, PIRO, SAH and Sn Coloidal methods use ITLC-SG; IPEN uses this chromatography plate in DISIDA, EC, ECD, GHA, PIRO, MIBI and SAH. As ITLC-SG had been out of production (recommended in 40, 70 and 41% of the USP, EP and IPEN methodologies, respectively), it was necessary to search alternatives to replace ITLC-SG plate in the radiochemical control

  18. Groundwater quality in wells in central rural Finland: a microbiological and radiochemical survey

    International Nuclear Information System (INIS)

    Korhonen, L.; Niskanen, M.; Heinonen-Tanski, H.; Martikainen, P.J.; Salonen, L.; Taipalinen, I.

    1996-01-01

    The microbiological, physicochemical, and radiochemical water quality from samples of 150 rural wells in Finland was analyzed. Organic matter exceeded 12 mg KMnO4 L(-1) in 63% and nitrate 25 mg NO3 L(-1) in 29% of the wells. NO3--concentrations were higher in wells with cattle. Fecal coliforms and fecal streptococci were found in 10-40%. There was no direct positive correlation between heterotrophic and indicator bacteria. Salmonella or Campylobacter were not detected. Human pathogen Listeria monocytogenes was isolated from two and Yersinia enterocolitica serotypes O5 or O6 from four waters not containing fecal coliforms. Thus, the predictive value of fecal coliforms to indicate these pathogens is poor. Coliphages were found in seven wells. Mean concentrations of radon and long-lived alpha-active radionuclides were lower and those of beta-emitting radionuclides higher than the mean concentrations measured from groundwater in Finland. Radionuclides from the Chernobyl fallout were not detected

  19. Standard practices for dissolving glass containing radioactive and mixed waste for chemical and radiochemical analysis

    CERN Document Server

    American Society for Testing and Materials. Philadelphia

    2000-01-01

    1.1 These practices cover techniques suitable for dissolving glass samples that may contain nuclear wastes. These techniques used together or independently will produce solutions that can be analyzed by inductively coupled plasma atomic emission spectroscopy (ICP-AES), inductively coupled plasma mass spectrometry (ICP-MS), atomic absorption spectrometry (AAS), radiochemical methods and wet chemical techniques for major components, minor components and radionuclides. 1.2 One of the fusion practices and the microwave practice can be used in hot cells and shielded hoods after modification to meet local operational requirements. 1.3 The user of these practices must follow radiation protection guidelines in place for their specific laboratories. 1.4 Additional information relating to safety is included in the text. 1.5 The dissolution techniques described in these practices can be used for quality control of the feed materials and the product of plants vitrifying nuclear waste materials in glass. 1.6 These pr...

  20. Radiochemical applications of insoluble sulfate columns. Analytical possibilities in the field of the fission product solutions

    International Nuclear Information System (INIS)

    Barrachina, M.; Sauvagnac, R.

    1962-01-01

    In this paper we go on with our study of the heterogeneous ion-isotopic exchange in column. At present, we apply it to determine the radiochemical composition of the raw solutions used in the industrial recuperation of the long-lived fission products. The separation of the radioelements contained in these solutions is carried out mainly by making use of small columns, 1-3 cm height, of BaSO 4 or SrSO 4 , under selected experimental conditions. These columns behave like a special type of inorganic exchangers, working by absorption or by ion-isotopic exchange depending on the cases,a nd they provide the means for the selective separation of several important fission products employing very small volumes of fixing and eluting solutions. (Author) 11 refs

  1. Assessment of radiochemical purity of [{sup 18}F]fludeoxyglucose by high pressure liquid chromatography (HPLC)

    Energy Technology Data Exchange (ETDEWEB)

    Lacerda, Aline E.; Silva, Juliana B.; Silveira, Marina B.; Ferreira, Soraya Z., E-mail: radiofarmacoscdtn@cdtn.b [Centro de Desenvolvimento da Tecnologia Nuclear (CDTN/CNEN-MG), Belo Horizonte, MG (Brazil). Unidade de Pesquisa e Producao de Radiofarmacos

    2011-07-01

    The quality control of [{sup 18}F]fludeoxyglucose ({sup 18}FDG) has received attention due to its increasing clinical use. Although the quality requirements of {sup 18}FDG are established in various pharmacopoeia, the suitability of all testing methods used should be verified under actual conditions of use and documented. The aim of this study was to develop a high pressure liquid chromatography (HPLC) method for radiochemical purity evaluation of {sup 18}FDG, based on pharmacopoeia references, and to verify its suitability for routine quality control in our centre. HPLC analysis was performed with an Agilent HPLC. {sup 18}FDG and impurities were separated on an anion-exchange column by isocratic elution with 0.1 M NaOH as the mobile phase. Detection was accomplished with refractive index and NaI (Tl) scintillation detectors. The flow rate of the mobile phase was set at 0.8 mL/min and the column temperature was kept at 35 deg C. Specificity, linearity, precision and robustness were assessed to verify if the method was adequate for its intended purpose. Retention time of {sup 18}FDG was not affected by the presence of other components of the formulation and a good peak resolution was achieved. The analytical curve of {sup 18}FDG was linear, with a correlation coefficient value of 0.9995. Intraday repeatable precision, reported as the relative standard deviation, was 0.11%. Analytical procedure remained unaffected by small variations in mobile phase flow rate. Results evidenced that HPLC is suitable for radiochemical purity evaluation of {sup 18}FDG, considering operational conditions of our laboratory. (author)

  2. Radiochemical neutron activation analysis of zirconium and zirconium-niobium alloys

    International Nuclear Information System (INIS)

    Tashimova, F.A.; Sadikov, I.I.; Salimov, M.

    2004-01-01

    Full text: Zirconium and zirconium-niobium alloys are used on nuclear technology, as fuel cladding of nuclear reactors. Their nuclear-physical, mechanical and thermophysical properties are influenced them matrix and impurity composition, therefore determination of matrix and impurity content of these materials is a very important task. Neutron activation analysis is one from multielemental and high sensible techniques that are widely applied in analysis of high purity materials. Investigation of nuclear-physical characteristics of zirconium has shown that instrumental variant NAA is unusable for analysis due to high radioactivity of a matrix. Therefore it is necessary carrying out radiochemical separation of impurity radionuclides from matrix. Study of the literature datum have shown, that zirconium and niobium are very well extracted from muriatic solution with 5% tributyl phosphineoxide (TBPO) solution in toluene and 0,75 M solution of di-2-ethyl hexyl phosphoric acid (HDEHP) in cyclohexanone. Investigation of these elements extraction in these systems has shown that more effective and selective separation of matrix radionuclides is achieved in HDEHP-3M HCI system. This system is also extracted and hafnium, witch is an accompanying element of zirconium and its high content prevented determination of other impurity elements in sample. Therefore we used extraction system HDEHP-3M HCl for analysis of zirconium and zirconium-niobium alloys in chromatographic variant. By measurement of distribution profile of a matrix and of elution curve of determined elements is established, that for effective separation of impurity and matrix radionuclides there is enough chromatographic column with diameter 1 cm and height of a sorbent layer 7 cm, thus volume of elute, necessary for complete elution of determinate elements is 35-40 ml. On the basis of the carried out researches the technique of radiochemical NAA of high purity zirconium and zirconium-niobium alloy, which allows to

  3. On-Line Monitoring for Control and Safeguarding of Radiochemical Streams at Spent Fuel Reprocessing Plant

    International Nuclear Information System (INIS)

    Bryan, Samuel A.; Levitskaia, Tatiana G.; Billing, Justin M.; Casella, Amanda J.; Johnsen, Amanda M.; Peterson, James M.

    2009-01-01

    Advanced techniques enabling enhanced safeguarding of the spent fuel reprocessing plants are urgently needed. Our approach is based on prerequisite that real time monitoring of the solvent extraction flowsheets provides unique capability to quickly detect unwanted manipulations with fissile isotopes present in the radiochemical streams during reprocessing activities. The methods used to monitor these processes must be robust and must be able to withstand harsh radiation and chemical environments. A new on-line monitoring system satisfying these requirements and featuring Raman spectroscopy combined with a Coriolis and conductivity probes, has been recently developed by our research team. It provides immediate chemical data and flow parameters of high-level radioactive waste streams with high brine content generated during retrieval activities from Hanford nuclear waste storage tanks. The nature of the radiochemical streams at the spent fuel reprocessing plant calls for additional spectroscopic information, which can be gained by the utilization of UV-vis-NIR capabilities. Raman and UV-vis-NIR spectroscopies are analytical techniques that have extensively been extensively applied for measuring the various organic and inorganic compounds including actinides. The corresponding spectrometers used under the laboratory conditions are easily convertible to the process-friendly configurations allowing remote measurements under the flow conditions. A fiber optic Raman probe allows monitoring of the high concentration species encountered in both aqueous and organic phases within the UREX suite of flowsheets, including metal oxide ions, such as uranyl, components of the organic solvent, inorganic oxo-anions, and water. The actinides and lanthanides are monitored remotely by UV-vis-NIR spectroscopy in aqueous and organic phases. In this report, we will present our recent results on spectroscopic measurements of simulant flowsheet solutions and commercial fuels available at

  4. Project Title: Radiochemical Analysis by High Sensitivity Dual-Optic Micro X-ray Fluorescence

    International Nuclear Information System (INIS)

    Havrilla, George J.; Gao, Ning

    2002-01-01

    A novel dual-optic micro X-ray fluorescence instrument will be developed to do radiochemical analysis of high-level radioactive wastes at DOE sites such as Savannah River Site and Hanford. This concept incorporates new X-ray optical elements such as monolithic polycapillaries and double bent crystals, which focus X-rays. The polycapillary optic can be used to focus X-rays emitted by the X-ray tube thereby increasing the X-ray flux on the sample over 1000 times. Polycapillaries will also be used to collect the X-rays from the excitation site and screen the radiation background from the radioactive species in the specimen. This dual-optic approach significantly reduces the background and increases the analyte signal thereby increasing the sensitivity of the analysis. A doubly bent crystal used as the focusing optic produces focused monochromatic X-ray excitation, which eliminates the bremsstrahlung background from the X-ray source. The coupling of the doubly bent crystal for monochromatic excitation with a polycapillary for signal collection can effectively eliminate the noise background and radiation background from the specimen. The integration of these X-ray optics increases the signal-to-noise and thereby increases the sensitivity of the analysis for low-level analytes. This work will address a key need for radiochemical analysis of high-level waste using a non-destructive, multi-element, and rapid method in a radiation environment. There is significant potential that this instrumentation could be capable of on-line analysis for process waste stream characterization at DOE sites

  5. Assessment of radiochemical purity of [18F]fludeoxyglucose by high pressure liquid chromatography (HPLC)

    International Nuclear Information System (INIS)

    Lacerda, Aline E.; Silva, Juliana B.; Silveira, Marina B.; Ferreira, Soraya Z.

    2011-01-01

    The quality control of [ 18 F]fludeoxyglucose ( 18 FDG) has received attention due to its increasing clinical use. Although the quality requirements of 18 FDG are established in various pharmacopoeia, the suitability of all testing methods used should be verified under actual conditions of use and documented. The aim of this study was to develop a high pressure liquid chromatography (HPLC) method for radiochemical purity evaluation of 18 FDG, based on pharmacopoeia references, and to verify its suitability for routine quality control in our centre. HPLC analysis was performed with an Agilent HPLC. 18 FDG and impurities were separated on an anion-exchange column by isocratic elution with 0.1 M NaOH as the mobile phase. Detection was accomplished with refractive index and NaI (Tl) scintillation detectors. The flow rate of the mobile phase was set at 0.8 mL/min and the column temperature was kept at 35 deg C. Specificity, linearity, precision and robustness were assessed to verify if the method was adequate for its intended purpose. Retention time of 18 FDG was not affected by the presence of other components of the formulation and a good peak resolution was achieved. The analytical curve of 18 FDG was linear, with a correlation coefficient value of 0.9995. Intraday repeatable precision, reported as the relative standard deviation, was 0.11%. Analytical procedure remained unaffected by small variations in mobile phase flow rate. Results evidenced that HPLC is suitable for radiochemical purity evaluation of 18 FDG, considering operational conditions of our laboratory. (author)

  6. Characterization of filter cartridges from the IEA-R1 reactor by radiochemical method

    International Nuclear Information System (INIS)

    Geraldo, Bianca; Vicente, Roberto; Ferreira, Robson J.; Goes, Marcos M.; Marumo, Julio T.

    2015-01-01

    The filter cartridges used in water purification system of research nuclear reactor IEA-R1 are considered radioactive wastes after their useful life. The characterization of these wastes is one of the stages of management, which aims to identify and quantify the radionuclides present, including those known as 'difficult to measure' (DTM) radionuclides. Establish a radiochemical analysis methodology for this type of waste is a difficult job, not only by the application of these techniques, but also by the amount of radionuclides that should be analyzed. In the waste produced in a nuclear reactor, the most important radionuclides are fission products, activation products and transuranic elements. Since these radionuclides emit gamma radiation not measurable in its decay process and consequently are difficult to measure, their concentrations can be estimated by indirect methods such as scale factors. This method is used to evaluate the DTM concentration, which is represented by alpha and beta nuclides using the correlation between them and the radionuclide key, a gamma emitter. The objective of this work is to describe a radiochemical analysis methodology for gamma emitter nuclides, present in the filter cartridges, evaluating the activity and concentrations by destructive assays. At the same time, two studies have been performed by non-destructive assays, the first one based on dose rates and the point kernel method to correlate the results and the second one based on calibration efficiency with Monte Carlo method. These studies belong to the radioactive waste characterization program that has been conducted at the Waste Management Laboratory of Nuclear and Energy Research Institute, IPEN-CNEN/SP. (author)

  7. The reliability of radiochemical and chemical trace analyses in environmental materials

    International Nuclear Information System (INIS)

    Heinonen, Jorma.

    1977-12-01

    After theoretically exploring the factors which influence the quality of analytical data as well as the means by which a sufficient quality can be assured and controlled, schemes of different kinds have been developed and applied in order to demonstrate the analytical quality assurance and control in practice. Methods have been developed for the determination of cesium, bromine and arsenic by neutron activation analysis at the natural ''background'' concentration level in environmental materials. The calibration of methods is described. The methods were also applied on practical routine analysis, the results of which are briefly reviewed. In the case of Ce the precision of a comprehensive calibration was found to vary between 5.2-9.2% as a relative standard deviation, which agrees well with the calculated statistical random error 5.7-8.7%. In the case of Br the method showed a reasonable precision, about 11% on the average, and accuracy. In employing the method to analyze died samples containing Br from 3 to 12 ppm a continuous control of precison was performed. The analysis of As demonstrates the many problems and difficulties associated with environmental analysis. In developing the final method four former intercomparison materials of IAEA were utilized in the calibration. The tests performed revealed a systematic error. In this case a scheme was developed for the continuous control of both precision and accuracy. The results of radiochemical analyses in environmental materials show a reliability somewhat better than that occuring in the determination of stable trace elements. According to a rough classification, 15% of the results of radiochemical analysis show excellent reliability, whereas 60% show a reliability adequate for certain purposes. The remaining 15% are excellent, 60% adequate for some purposes and 30% good-for-nothing. The reasons for often insufficient reliability of results are both organizational and technical. With reasonable effort and

  8. Improving the radiochemical purity determination of 123I-labeled metaiodobenzylguanidine

    International Nuclear Information System (INIS)

    Luciana Carvalheira; Paulo Bechara Dutra; Paula Fernandes de Aguiar

    2014-01-01

    The HPLC method originally applied at the Nuclear Engineering Institute (IEN) for the radiochemical purity determination of 123iodine labeled m-iodobenzylguanidine ( 123 I-mIBG) takes 18.5 min. The final product release also depends on this result, and to facilitate this stage, we aimed to decrease this analysis time. We also intended to use fewer toxic compounds, if feasible. The optimization approach used herein was a combination of factorial and mixture designs to study simultaneously the selected variables. Analysis time, resolution and chromatograms aspect were the measured responses. The qualitative analysis of these responses provided the best chromatographic separation conditions that were 52 mM KH 2 PO 4 in a solution of ethanol and water (1:1), applying a flow rate of 0.50 mL min -1 and C18 column (4.6 × 250 mm, 5 μm). These optimum conditions not only decreased the analysis time in 61 %, but also allowed the reduction of mobile phase toxicity. To assure reliable data, method validation was performed for these conditions. The method has proved its specificity, the detection limit found was 3.70 × 10 -4 MBq mL -1 and the quantification limit has corresponded to 1.11 × 10 -3 MBq mL -1 . Repeatability and intermediate precision has not exceeded 3 and 5 %, respectively, and the accuracy has matched the interval of 95-105 %. This new method has been routinely applied in the radiochemical purity determination of 123 I-mIBG at IEN. (author)

  9. Characterization of filter cartridges from the IEA-R1 reactor by radiochemical method

    Energy Technology Data Exchange (ETDEWEB)

    Geraldo, Bianca; Vicente, Roberto; Ferreira, Robson J.; Goes, Marcos M.; Marumo, Julio T., E-mail: bgeraldo@ipen.br [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil)

    2015-07-01

    The filter cartridges used in water purification system of research nuclear reactor IEA-R1 are considered radioactive wastes after their useful life. The characterization of these wastes is one of the stages of management, which aims to identify and quantify the radionuclides present, including those known as 'difficult to measure' (DTM) radionuclides. Establish a radiochemical analysis methodology for this type of waste is a difficult job, not only by the application of these techniques, but also by the amount of radionuclides that should be analyzed. In the waste produced in a nuclear reactor, the most important radionuclides are fission products, activation products and transuranic elements. Since these radionuclides emit gamma radiation not measurable in its decay process and consequently are difficult to measure, their concentrations can be estimated by indirect methods such as scale factors. This method is used to evaluate the DTM concentration, which is represented by alpha and beta nuclides using the correlation between them and the radionuclide key, a gamma emitter. The objective of this work is to describe a radiochemical analysis methodology for gamma emitter nuclides, present in the filter cartridges, evaluating the activity and concentrations by destructive assays. At the same time, two studies have been performed by non-destructive assays, the first one based on dose rates and the point kernel method to correlate the results and the second one based on calibration efficiency with Monte Carlo method. These studies belong to the radioactive waste characterization program that has been conducted at the Waste Management Laboratory of Nuclear and Energy Research Institute, IPEN-CNEN/SP. (author)

  10. Development of a radiochemical neutron activation analysis procedure for determination of rhenium in biological and environmental samples at ultratrace level

    Czech Academy of Sciences Publication Activity Database

    Kučera, Jan; Byrne, A. R.; Mizera, Jiří; Lučaníková, M.; Řanda, Zdeněk

    2006-01-01

    Roč. 269, č. 2 (2006), s. 251-257 ISSN 0236-5731 R&D Projects: GA ČR(CZ) GA203/04/0943 Institutional research plan: CEZ:AV0Z10480505 Keywords : radiochemical neutron activation analysis * rhenium * biological and environmental samples Subject RIV: CB - Analytical Chemistry, Separation Impact factor: 0.509, year: 2006

  11. Obtention process of phosphorus 32 starting from commercial sulfur and design and construction of the radiochemical separation prototype

    International Nuclear Information System (INIS)

    Duarte A, C.; Alanis M, J.; Gutierrez R, C.

    2002-01-01

    In this work an obtention process of phosphorus 32 ( 32 P) in orthophosphoric acid form (H 3 32 PO 4 ) is described starting from commercial sulfur. Also the design and construction of the experimental prototype used in the radiochemical separation and their results in three tests carried out is reported. (Author)

  12. Radiochemical separations of hafnium, tantalum, and germanium from tungsten and selenium irradiated by 14 MeV neutrons

    International Nuclear Information System (INIS)

    Blachot, J.; Benabed, A.; Herment, J.; Monnand, E.

    1968-01-01

    Radiochemical separations of Hf, Ta and Ge from W and Se respectively, after 14 MeV-neutron irradiation. A new isotope: 79 Ge with a half-live of 50 ± 5 s and emitter of a 230 keV γ transition is observed. (authors) [fr

  13. Radiochemical and thermal studies of the copper(II)-exchanged form of synthetic zeolite linde sieve A

    International Nuclear Information System (INIS)

    Banerjee, S.P.

    1978-01-01

    Synthetic zeolite Linde Sieve A displays a double ion-sieve action. Only small cations can penetrate the single 6-rings into the beta cages. The radiochemical and thermal studies of copper(II)-exchanges form of 4A shows evidence of hydrated copper(II) ions in the zeolite structure. (author)

  14. System-wide Analysis of SUMOylation Dynamics in Response to Replication Stress Reveals Novel Small Ubiquitin-like Modified Target Proteins and Acceptor Lysines Relevant for Genome Stability

    DEFF Research Database (Denmark)

    Xiao, Zhenyu; Chang, Jer-Gung; Hendriks, Ivo A

    2015-01-01

    . Following statistical analysis on five biological replicates, a total of 566 SUMO-2 targets were identified. After 2 hours of Hydroxyurea treatment, 10 proteins were up-regulated for SUMOylation and 2 proteins were down-regulated for SUMOylation, whereas after 24 hours, 35 proteins were up...

  15. On the presence of fictitious solar neutrino flux variations in radiochemical experiments

    International Nuclear Information System (INIS)

    Vladimirskii, B.M.; Bruns, A.V.

    2004-01-01

    The currently available data on solar neutrino flux variation in radiochemical experiments and Cherenkov measurements have so far defied a simple interpretation. Some of the results concerning these variations are indicative of their relationship to processes on the solar surface. It may well be that a poorly understood, uncontrollable factor correlating with solar activity indices affects the neutrino flux measurements. This factor is assumed to modulate the detection efficiency on different detectors in different ways. To test this assumption, we have analyzed all available radiochemical measurements obtained with the Brookhaven (1970-1994, 108 runs), GALLEX (1991-1997, 65 runs), and SAGE (1989-2000, 80 runs) detectors for possible instability of the detection efficiency. We consider the heliophysical situation at the final stage of the run, the last 7-27 days, when the products of the neutrino reaction with the target material had already been accumulated. All of the main results obtained previously by other authors were found to be reproduced for chlorine-argon measurements. The neutrino flux anticorrelates with the sunspot numbers only for an odd solar cycle. A similar behavior is observed for the critical frequencies of the E-ionosphere. The neutrino flux probably correlates with the A p magnetic activity index only for an even solar cycle. The predominance of a certain sign of the radial interplanetary magnetic field (IMF) in the last 14 (or 7) days of the run has the strongest effect on the recorded neutrino flux. The effect changes sign when the polarity of the general solar magnetic field is reversed and is most pronounced for the shortest runs (less than 50 days). The dependence of the flux on IMF polarity completely disappears if the corresponding index is taken for the first rather than the last days of the run. The IMF effect on the recorded neutrino flux was also found for short runs in the gallium-germanium experiment, but this effect for a given

  16. Investigation of Chemical and Radiochemical Fingerprints of Water Resources in Siwa Oasis, Western Desert, Egypt

    International Nuclear Information System (INIS)

    El-Sayed, S.A.; Allam, Kh.A.; Salama, M.H.; El Begawy, H.

    2017-01-01

    Water resources in Siwa oasis, Egypt; included the Nubian sandstone aquifer system (NSSAS), Tertiary carbonate aquifer system (TCAS) as well as springs and lakes. Existing chemical and radiochemical fingerprints of these resources in the central and western parts of the oasis were examined based on the analyses of 30 water samples collected in 2015. In each sample, the analyzed chemical and radioisotopic constituents were Cl"-, SO_4"2"- , CO_3"2"-, HCO"3 "-, Na"+, K"+, Ca"2"+ and Mg"2"+, Al, B, Ba, Cd, Co, Fe, Cu, Mn, Mo, Ni, Pb, Sr, V, Zn, Si and Cr, "2"2"6 Ra, "2"3"2Th and "4"0K. Other physical properties included ph and TDS were measured in the field. The chemical analysis was carried out following the ASTM methods, while the radiochemical measurements were done using a hyper pure germanium (HPGe) gamma ray spectrometry. Results of this work may be advantage ous to promote the hydrologic database in the oasis, where levels of the natural radioactivity in groundwater resources were established for the first time. Water resources were classified based on their contents of TDS into fresh water for the NSSAS, brackish to salty water for TCAS, brackish water for springs and brine water for lakes. The artesian water of the NSSAS and TCAS and water of the springs were alkaline, while the water of the lakes was acidic. Chemical characters of the ground water of the NSSAS reflected a little response to the processes of water-rock interactions, and the vice versa was observed for the groundwater of the TCAS. The ground water of the NSSAS was dominated by HCO"3"-Na genetic type characterizing the meteoric origin of water. The ground water of the TCAS was characterized by the presence of three genetic types of water (SO"4"-Na, Cl-Mg, and Cl-CA water), indicating the modification of the recharged water from the underlain NSSAS due to the effects of water sediment interactions and mixing with trapped ancient sea water. The chemical properties of spring water were, greatly

  17. Application of radiochemical methods for development of new biological preparation designed for soil bioremediation

    International Nuclear Information System (INIS)

    Kim, A.A.; Djuraeva, G.T.; Djumaniyazova, G.I.; Yadgarov, Kh.T.

    2006-01-01

    Full text: Internationally the bioremediation of agricultural lands contaminated by persistent chloroorganic compounds by means of the microbial methods are used as the most low-cost and the most effective. One of the factors reducing efficacy of microbial degradation, is often the low quantity of microorganisms - destructors in the soil. Therefore, we have designed bioremediation technology of soils, contaminated by organochlorine compounds, with use of the alive microorganisms as active agent. We developed the biological preparation containing 5 aboriginal active strains of bacteria - destructors of persistent chloroorganic compounds and investigated the ability of biological preparation to increase the bioremediation potential of contaminated soils. To carry out the investigation we developed the complex of radiochemical methods with use of tritium labeled PCBs, including the following methods: 1.The method to define the accumulation and degradation of PCBs in soil bacteria in culture allows determination of quantitative characteristics of bacterial strains. 2. The method to define the PCBs degradation by soil bacteria strains in model conditions in the soil allows to estimate the PCB-destructive activity of strains after introducing in soil. 3. A method to define the PCB-destructive activity of own microbiota of contaminated soil. 4. A method to define the effect of stimulation of the PCB-destructive activity of biological preparation and own microbiota of soil with the help of biofertilizers. By using the developed radiochemical methods we have carried out investigation on creation of new biological preparation on the basis of strains of soil bacteria - destructors of PCBs. We also determined the quality and quantity characteristics of HCCH and PCBs-destructive activity of new biological preparation. It is shown that the new biological preparation is capable of accumulation and destruction of the PCBs in culture and in soil at model conditions. Thus, the

  18. The eIF4E-binding proteins are modifiers of cytoplasmic eIF4E relocalization during the heat shock response

    OpenAIRE

    Sukarieh, R.; Sonenberg, N.; Pelletier, J.

    2009-01-01

    Stress granules (SGs) arise as a consequence of cellular stress, contain stalled translation preinitiation complexes, and are associated with cell survival during environmental insults. SGs are dynamic entities with proteins relocating into and out of them during stress. Among the repertoire of proteins present in SGs is eukaryotic initiation factor 4E (eIF4E), a translation factor required for cap-dependent translation and that regulates a rate-limiting step for protein synthesis. Herein, we...

  19. III Scientific-technical conference. Problems and outlooks for development of chemical and radiochemical control in atomic energetics (Atomenergoanalytics-2005). Summaries of reports

    International Nuclear Information System (INIS)

    2005-01-01

    Summaries of reports of the III Scientific-technical conference: Problems and outlooks for development of chemical and radiochemical control in atomic energetics (Atomenergoanalytics-2005) are presented. The conference performed 20-22 September, 2005, in Sosnovyj Bor. Problems of methodical, instrumental and metrological supply of chemical, radiochemical and radiometric control at active NPP and NPU, modern concepts of construction of automated systems of chemical and radiometric control in the atomic energetics, directions for the decision of questions in organization and conducting of chemical and radiochemical control of water-chemical regimes of NPP and NPU are discussed [ru

  20. Radiochemical neutron activation analysis of high pure palladium and platinum by ion exchange chromatography

    International Nuclear Information System (INIS)

    Sadikov, I.I.; Zinov'ev, V.G.; Sadikova, Z.O.; Salimov, M.I.

    2006-01-01

    Full text: The palladium and platinum are widely used for jewel manufacture because of their beautiful white color. However the most part of these metals are widely adopted in the world as catalysts. Many works on analytical chemistry of platinum group elements published during last years are devoted to determination of platinum and palladium in other materials. There are no articles on analysis technique of the palladium and platinum purity published during last 20 years. Available publications are very old and are published till 70th of the last century, and implement chemical and spectral methods. At the same time, the palladium and platinum are very suitable for NAA. Therefore the purpose of our research was development of high-sensitivity and multielement techniques of radiochemical neutron activation analysis of a high pure palladium and platinum. Research of nuclear characteristics of palladium and platinum has shown that radioactive nuclides with different yields are formed under the reactor neutrons. 109 , 111 , 111m Pd, 109m , 111 Ag, 191 , 197 , 199 Pt, 199 Au are the most important among them. 109Pd separation factor is equal to 1*10 5 at palladium analysis, whereas 197 Pt and 199 Au separation factor is equal to 1*10 4 at the platinum analysis every other day after irradiation. Palladium and platinum can be separated by precipitation, extraction and ion exchange methods. For separation of radioactive nuclide of the matrix elements from the impurity elements we used ion exchange chromatography system Dowex-1x8 - 1 M HNO 3 for palladium and Dowex-1x8 - 0.1 M HNO 3 for platinum. At the HNO 3 acid concentrations variation from 0,1 M to 1 M more then 25 elements have distribution factors less than 1 and 10 elements have distribution factors 5 while matrix elements have distribution factors higher than 100. It allows an effective separation of these elements from palladium and platinum. Optimum sizes of the chromatographic column and the column effluent