WorldWideScience

Sample records for isol method development

  1. Development of analysis methods for seismically isolated nuclear structures

    International Nuclear Information System (INIS)

    Yoo, Bong; Lee, Jae-Han; Koo, Gyeng-Hoi

    2002-01-01

    KAERI's contributions to the project entitled Development of Analysis Methods for Seismically Isolated Nuclear Structures under IAEA CRP of the intercomparison of analysis methods for predicting the behaviour of seismically isolated nuclear structures during 1996-1999 in effort to develop the numerical analysis methods and to compare the analysis results with the benchmark test results of seismic isolation bearings and isolated nuclear structures provided by participating countries are briefly described. Certain progress in the analysis procedures for isolation bearings and isolated nuclear structures has been made throughout the IAEA CRPs and the analysis methods developed can be improved for future nuclear facility applications. (author)

  2. Methods Development for the Isolation and Culture of Primary Corneal Endothelial Cells

    Science.gov (United States)

    2017-02-01

    a cell population particularly suitable for low serum propagation, provided that appropriate growth factors are available. A low serum medium...of MGK. 15. SUBJECT TERMS Cornea, chemical warfare agent, corneal endothelial cell, endothelium, growth , isolation, mouse, rabbit, porcine, in...with corneal SM exposure.2 A primary requirement in achieving this goal is to develop methods that enable the isolation of a pure CEC population and

  3. Isolation and HPLC method development of azafrin from Alectra parasitica var. chitrakutensis.

    Science.gov (United States)

    Agrawal, Poonam; Laddha, Kirti; Tiwari, Ashok

    2014-01-01

    This study was undertaken to isolate and quantify azafrin in Alectra parasitica (Scrophulariaceae) rhizomes. A simple method for the isolation of carotenoid, azafrin, involves solvent extraction of the dried rhizome powder using a single solvent and further purification by recrystallisation. The structure of the compound was elucidated and confirmed by thin-layer chromatography, infrared spectroscopy, mass spectroscopy and nuclear magnetic resonance spectral analysis. A specific and rapid reversed-phase high-performance liquid chromatography (HPLC) method was developed for the analysis of azafrin. The method was validated for accuracy, precision, linearity and specificity. Validation revealed that the method is specific, accurate, precise, reliable and reproducible. The proposed HPLC method can be used for the identification and quantitative analysis of azafrin in A. parasitica rhizomes.

  4. Development of an evaluation method for seismic isolation systems of nuclear power facilities. Development of crossover piping design method for seismic isolation systems

    International Nuclear Information System (INIS)

    Otoyo, Teruyoshi; Otani, Akihito; Otani, Akihito; Fukushima, Shunsuke; Jimbo, Masakazu; Yamamoto, Tomofumi; Sakakida, Takaaki; Onishi, Shigenobu

    2014-01-01

    In the conceptual design of seismic isolation systems of nuclear power facilities, there exist two types of installation. The first type is to isolate both the reactor and the turbine buildings, the other is to isolate only the reactor building. In the latter type, the crossover piping, which installed between the isolated and the non-isolated buildings, is excited and deformed by the different motions of those buildings. In this study, shaking tests of 1/10 scaled model of the main steam piping and FEM analyses under multiple support excitation conditions have been performed to investigate the vibration behavior of the crossover piping. It was confirmed that modal time-history analyses could be in good agreement with the shaking test results. Also, Numerous combination methods were investigated by comparing response spectrum analyses and modal time-history analyses. In conclusion, response spectrum analyses using SRSS combinations could correspond to time-history analyses. (author)

  5. Development of design method of thick rubber bearings for three-dimensional base isolation

    International Nuclear Information System (INIS)

    Yabana, Shuichi; Matuda, Akihiro

    2000-01-01

    Thick rubber bearings as 3-dimensional base isolators have been developed to reduce both horizontal and vertical seismic loads especially for equipment in Fast Breeder Reactors. In this report, a design method of thick rubber bearings is presented. To consider nonlinearity of vertical stiffness affected by vertical stress in the design of thick rubber bearings, Lindley's evaluation method of vertical stiffness is modified as an explicit form of vertical stress. We confirm that the presented method is efficient for design of the thick rubber bearings from comparing between test results and predicted values. Furthermore, rubber bearing tests are conducted with 1/3 scale models to evaluate mechanical properties of thick rubber bearings including ultimate limits. In the tests, horizontal and vertical characteristics of 1/3 scale model are compared with those of 1/6 scale model to discuss scale effect of test specimen. Ultimate limits such as failure shear strain of thick rubber bearings are obtained under various loading conditions. From the test results, we confirm that full scale thick rubber bearing to satisfy requirements is feasible. (author)

  6. A Method for Recruiting Participants from Isolated Islands of Small Island Developing States (SIDS) for Survey Research

    Science.gov (United States)

    Moosa, Sheena; Koopman-Boyden, Peggy

    2016-01-01

    Representing isolated small island communities through social survey research continues to be challenging. We examine a locally developed method to reach and recruit older people (65+ years) for a survey on well-being in the small island developing state of Maldives. The use of messengers to recruit participants is examined in the context of these…

  7. Schizosaccharomyces isolation method

    Directory of Open Access Journals (Sweden)

    Benito Santiago

    2014-01-01

    Full Text Available This study discusses the optimization of a selective and differential medium which would facilitate the isolation of Schizosaccharomyces (a genus with a low incidence compared to other microorganisms to select individuals from this genus for industrial purposes, especially in light of the recent recommendation of the use of yeasts from this genus in the wine industry by the International Organisation of Vine and Wine, or to detect the presence of such yeasts, for those many authors who consider them food spoilers. To this end, we studied various selective differential agents based on the main physiological characteristics of these species, such as their high resistances to high concentrations of sugar, sulfur dioxide, sorbic acid, benzoic acid, acetic acid or malo ethanolic fermentation. This selective medium is based on the genus resistance to the antibiotic actidione and its high resistance to inhibitory agents such as benzoic acid. Malic acid was used as a differential factor due to the ability of this genus to metabolise it to ethanol, which allows detecting of the degradation of this compound. Lastly, the medium was successfully used to isolate strains of Schizosaccharomyces pombe from honey and honeycombs.

  8. Development of a Method to Isolate Glutamic Acid from Foodstuffs for a Precise Determination of Their Stable Carbon Isotope Ratio.

    Science.gov (United States)

    Kobayashi, Kazuhiro; Tanaka, Masaharu; Yatsukawa, Yoichi; Tanabe, Soichi; Tanaka, Mitsuru; Ohkouchi, Naohiko

    2018-01-01

    Recent growing health awareness is leading to increasingly conscious decisions by consumers regarding the production and traceability of food. Stable isotopic compositions provide useful information for tracing the origin of foodstuffs and processes of food production. Plants exhibit different ratios of stable carbon isotopes (δ 13 C) because they utilized different photosynthetic (carbon fixation) pathways and grow in various environments. The origins of glutamic acid in foodstuffs can be differentiated on the basis of these photosynthetic characteristics. Here, we have developed a method to isolate glutamic acid in foodstuffs for determining the δ 13 C value by elemental analyzer-isotope-ratio mass spectrometry (EA/IRMS) without unintended isotopic fractionation. Briefly, following acid-hydrolysis, samples were defatted and passed through activated carbon and a cation-exchange column. Then, glutamic acid was isolated using preparative HPLC. This method is applicable to measuring, with a low standard deviation, the δ 13 C values of glutamic acid from foodstuffs derived from C3 and C4 plants and marine algae.

  9. Development of an evaluation method for seismic isolation systems of nuclear power facilities. Seismic design analysis methods for crossover piping system

    International Nuclear Information System (INIS)

    Tai, Koichi; Sasajima, Keisuke; Fukushima, Shunsuke; Takamura, Noriyuki; Onishi, Shigenobu

    2014-01-01

    This paper provides seismic design analysis methods suitable for crossover piping system, which connects between seismic isolated building and non-isolated building in the seismic isolated nuclear power plant. Through the numerical study focused on the main steam crossover piping system, seismic response spectrum analysis applying ISM (Independent Support Motion) method with SRSS combination or CCFS (Cross-oscillator, Cross-Floor response Spectrum) method has found to be quite effective for the seismic design of multiply supported crossover piping system. (author)

  10. Review of the ISOL Method

    CERN Document Server

    Lindroos, M

    2004-01-01

    The ISOL technique was invented in Copenhagen over 50 years ago and eventually migrated to CERN where a suitable proton drive beam was available at the Syncho-Cyclotron. The quick spread of the technique from CERN to many other laboratories has resulted in a large user community, which has assured the continued development of the method, physics in the front-line of fundamental research and the application of the method to many applied sciences. The technique is today established as one of the main techniques for on-line isotope production of high intensity and high quality beams. The thick targets used allows the production of unmatched high intensity radioactive beams. The fact that the ions are produced at rest makes it ideally suitable for low energy experiments and for post acceleration using well established accelerator techniques. The many different versions of the technique will be discussed and the many facilities spread all over the world will be reviewed. The major developments at the existing faci...

  11. Tiamulin activity against fastidious and nonfastidious veterinary and human bacterial isolates: initial development of in vitro susceptibility test methods.

    Science.gov (United States)

    Jones, Ronald N; Pfaller, Michael A; Rhomberg, Paul R; Walter, Donald H

    2002-02-01

    Tiamulin is a pleuromutilin derivative used in veterinary practice for the control and specific therapy of infections in swine. This report summarizes studies to establish standardized susceptibility testing methods, interpretive criteria, and reagent details for use in veterinary methods recently developed by the National Committee for Clinical Laboratory Standards (NCCLS) (standards M31-A and M37-A, NCCLS, Wayne, Pa., 1999). A total of 636 fastidious and nonfastidious animal and human pathogens were processed by using media and procedures described by the NCCLS. Tiamulin disk diffusion tests used a 30-microg disk concentration, and the proposed MIC breakpoints corresponding to levels achievable in animal target tissues (lung) were or =32 microg/ml for resistance. Correlate zone diameters for specific nonfastidious species were as follows: for Pasteurella multocida and staphylococci tested on Mueller-Hinton agar, susceptibility at > or =19 mm and resistance at or =16 mm and resistance at or =9 mm) was suggested for veterinary fastidious medium broth and enriched chocolate Mueller-Hinton agar. Absolute categorical agreement between NCCLS dilution and disk diffusion test results with these criteria ranged from 90.5 to 96.2%. Tiamulin susceptibility testing methods appear to be accurate in their categorical classification for indicated species, and their availability will allow immediate testing of animal isolates to guide therapy via appropriate levels of dosing and to monitor the development of resistance for agents in this unique class.

  12. Radioactive beams produced by the ISOL method: development for laser ionization and for surface ionization; Faisceaux exotiques par methode ISOL: developpements pour l'ionisation par laser et l'ionisation de surface

    Energy Technology Data Exchange (ETDEWEB)

    Hosni, Faouzi

    2004-10-01

    The works were carried out in the framework of the research program PARRNe (production of radioactive neutron-rich nuclei). This program aims to determine optimal conditions to produce intense beams of neutron-rich isotopes. This thesis treats multiple technical aspects related to the production of separate radioactive isotopes in line (ISOL). It deals mainly with the development of the target-source unit which is the key element for projects such as SPIRAL-2 or EURISOL.The first part presents the various methods using fission as production mode and compares them: fission induced by thermal neutrons, induced by fast neutrons and photofission. The experiment carried out at CERN validated the interest of the photofission as a promising production mode of radioactive ions. That is why the institute of nuclear physics of Orsay decided to build a linear electron accelerator at the Tandem d'Orsay (ALTO).The second part of this thesis deals with the development of uranium targets. The X-rays diffraction and Scanning Electron Microscopy have been used as analysis techniques. They allowed to determine the chemical and structural characteristics of uranium carbide targets as function of various heating temperatures. After the production, the process of ionization has been studied. Two types of ion source have been worked out: the first one is a surface ion source and the second one is a source based on resonant ionization by laser. These two types of sources will be used for the ALTO project. (author)

  13. K. OXYTOCA BACTERIOPHAGES ISOLATION METHODS IMPROVEMENT

    Directory of Open Access Journals (Sweden)

    G. R. Sadrtdinova

    2017-01-01

    Full Text Available The article presents the results of a study related to increasing the efficiency of phage isolation of bacteria of the species K. oxytoca, by developing the optimal composition of the medium used in the work. In scientific research, in almost all methods associated with the isolation of bacteriophages, meat-peptone broth and meat-peptone agar are used as the nutrient basis. The peculiarities of growth and cultivation of microorganisms create certain difficulties for the isolation of phages active against bacteria of the species K. oxytoca. The selection of components and the creation of an environment that would ensure the optimal growth of both the bacterial culture and the reproduction of the virus makes it possible to facilitate the isolation of bacteriophages. The number of bacterial strains used in the work was 7. All strains of cultures were obtained from the Museum of the Department of Microbiology, Virology, Epizootology and Veterinary and Sanitary Expertise of the Federal State Budget Educational Institution of Higher Education “Ulyanovsk State Agrarian University named after P.A. Stolypin”. The studies included 2 main stages. The first stage consisted in isolation of bacteriophages by the method of isolation from the external environment by the method of Adelson L.I., Lyashenko E.A. The material for the studies were samples: soil, sewage sample, fecal samples (2. Only 4 samples. According to the chosen method, the sowing of the putative phagolysate was carried out on meat-peptone agar (1.5% and the agar for isolating bacteriophages (Aph (1.5%. A positive result was the presence on the environment of negative colonies, clearly visible on the matt background of deep growth of bacteria. A negative result is a continuous growth (“lawn” of bacterial culture. As a control, the culture of the microorganism studied was used for the media. In the course of the conducted studies for the first stage, 2 bacteriophages were isolated, active

  14. Assessing Social Isolation: Pilot Testing Different Methods.

    Science.gov (United States)

    Taylor, Harry Owen; Herbers, Stephanie; Talisman, Samuel; Morrow-Howell, Nancy

    2016-04-01

    Social isolation is a significant public health problem among many older adults; however, most of the empirical knowledge about isolation derives from community-based samples. There has been less attention given to isolation in senior housing communities. The objectives of this pilot study were to test two methods to identify socially isolated residents in low-income senior housing and compare findings about the extent of isolation from these two methods. The first method, self-report by residents, included 47 out of 135 residents who completed in-person interviews. To determine self-report isolation, residents completed the Lubben Social Network Scale 6 (LSNS-6). The second method involved a staff member who reported the extent of isolation on all 135 residents via an online survey. Results indicated that 26% of residents who were interviewed were deemed socially isolated by the LSNS-6. Staff members rated 12% of residents as having some or a lot of isolation. In comparing the two methods, staff members rated 2% of interviewed residents as having a lot of isolation. The combination of self-report and staff report could be more informative than just self-report alone, particularly when participation rates are low. However, researchers should be aware of the potential discrepancy between these two methods.

  15. Development of EUCAST disk diffusion method for susceptibility testing of the Bacteroides fragilis group isolates

    DEFF Research Database (Denmark)

    Nagy, Elisabeth; Justesen, Ulrik Stenz; Eitel, Zsuzsa

    2015-01-01

    -clavulanic acid, cefoxitin, clindamycin, imipenem, metronidazole, moxifloxacin, piperacillin/tazobactam, tigecycline by agar dilution method previously. The inhibition zones of the same antibiotics including meropenem disc were determined by the disc diffusion on Brucella blood agar supplemented with haemin...

  16. Study on design method for seismically isolated FBR plants

    International Nuclear Information System (INIS)

    Hirata, Kazuta; Yabana, Shuichi; Ohtori, Yasuki; Ishida, Katsuhiko; Sawada, Yoshihiro; Shiojiri; Hiroo; Mazda, Taiji

    1998-01-01

    CRIEPI conducted 'Demonstration test on FBR seismic isolation system' from 1987 to 1996 under contract with Ministry of International Trade and Industry, Japan. In the demonstration test, base isolation technologies are prepared and demonstrated to apply to FBR and the design guidelines are proposed. In this report overall contents of the design guidelines entitled Design guidelines for seismically base isolated FBR plants' are included. The design guidelines, as a rule, are limited to apply to FBR plants where entire reactor building is isolated in the horizontal direction using laminated rubber bearings as isolators. The design guidelines and its concepts, however, will be useful for the development of similar guidelines for other isolation systems using different type of isolation methods and other nuclear facilities. The design guidelines consist of three parts and appendices. The first part is 'Policy for Safety Design of Base Isolated FBR Plants' specifying the principles and the requirements in the planning and the design for the safety of base isolated FBR plants. The second part is Policy for Seismic Design of Base Isolated FBR' describing the principles and the requirements in the seismic design and the evaluation of safety for base isolated FBR plants. The third part is 'Design Methods for Seismic Isolated FBR Plants' detailing the methods, procedures and parameters to be used in the design and the evaluation of safety fro base isolated FBR plants. In appendices examples of design procedures for base isolated reactor building and laminated rubber bearings as well as various test data on laminated rubber bearings, etc. are shown. (author)

  17. Optimized anion exchange column isolation of zirconium-89 (89Zr) from yttrium cyclotron target: Method development and implementation on an automated fluidic platform.

    Science.gov (United States)

    O'Hara, Matthew J; Murray, Nathaniel J; Carter, Jennifer C; Morrison, Samuel S

    2018-04-13

    Zirconium-89 ( 89 Zr), produced by the (p, n) reaction from naturally monoisotopic yttrium ( nat Y), is a promising positron emitting isotope for immunoPET imaging. Its long half-life of 78.4 h is sufficient for evaluating slow physiological processes. A prototype automated fluidic system, coupled to on-line and in-line detectors, has been constructed to facilitate development of new 89 Zr purification methodologies. The highly reproducible reagent delivery platform and near-real time monitoring of column effluents allows for efficient method optimization. The separation of Zr from dissolved Y metal targets was evaluated using several anion exchange resins. Each resin was evaluated against its ability to quantitatively capture Zr from a load solution high in dissolved Y. The most appropriate anion exchange resin for this application was identified, and the separation method was optimized. The method is capable of a high Y decontamination factor (>10 5 ) and has been shown to remove Fe, an abundant contaminant in Y foils, from the 89 Zr elution fraction. Finally, the method was evaluated using cyclotron bombarded Y foil targets; the method was shown to achieve >95% recovery of the 89 Zr present in the foils. The anion exchange column method described here is intended to be the first 89 Zr isolation stage in a dual-column purification process. Copyright © 2018 Elsevier B.V. All rights reserved.

  18. Methods to isolate extracellular vesicles for diagnosis

    Science.gov (United States)

    Kang, Hyejin; Kim, Jiyoon; Park, Jaesung

    2017-12-01

    Extracellular vesicles (EVs) are small membrane-bound bodies that are released into extracellular space by diverse cells, and are found in body fluids like blood, urine and saliva. EVs contain RNA, DNA and proteins, which can be biomarkers for diagnosis. EVs can be obtained by minimally-invasive biopsy, so they are useful in disease diagnosis. High yield and purity contribute to precise diagnosis of disease, but damaged EVs and impurities can cause confu sed results. However, EV isolation methods have different yields and purities. Furthermore, the isolation method that is most suitable to maximize EV recovery efficiency depends on the experimental conditions. This review focuses on merits and demerits of several types of EV isolation methods, and provides examples of how to diagnose disease by exploiting information obtained by analysis of EVs.

  19. Development of library preparation method able to correct gene expression levels in rice anther and isolate a trace expression gene mediated in cold-resistance

    International Nuclear Information System (INIS)

    Yamaguchi, Tomoya; Koike, Setsuo

    2000-01-01

    When cDNA library is prepared by a previously developed method, genes of which expression level is high are apt to be cloned at a high frequency, whereas genes of which expression level are low, are difficult to be cloned. A low-expression gene has been cloned at very low frequency. Therefore, the gene encoding the key enzyme that is involved in growth disturbance of rice pollen has not been identified. In this study, development of a library preparing method able to correct the expression level was attempted using highly sensitive detection method with radioisotope and some genes related to cold-resistance of rice were isolated. Double strand DNAs were synthesized using mRNA extract from rice anthers and annealed following heat-denaturation. It has been known that single strand DNA molecules abundantly existing in DNA solution can easily aggregate to form double strand DNA, but single stranded DNA molecules poor in the solution are apt to still remain as single strand after annealing. Thus, the amount of single strand DNA would be balanced in the solution between abundant DNA and poor DNA species. The authors succeeded to prepare a gene library including low and high expression genes at similar proportions. Moreover, spin trap method that allows RI labeling of DNA bound to latex particle, was developed to detect with high sensitivity, especially for genes that are expressed at low level. The present method could be used for recovery, detection and quantitative analysis of radiolabeled single strand DNA. Thus, it was demonstrated that the stage from tetrad sperm to small sperm might be easily affected by cold stress. The present results suggest that the expressions of β-1 and β-3 glucanase, which are involved in the release of small sperms following meiosis in the pollen formation, might be easily affected by cold stress. (M.N.)

  20. Development of library preparation method able to correct gene expression levels in rice anther and isolate a trace expression gene mediated in cold-resistance

    Energy Technology Data Exchange (ETDEWEB)

    Yamaguchi, Tomoya; Koike, Setsuo [Tohoku National Agricultural Experiment Station, Morioka (Japan)

    2000-02-01

    When cDNA library is prepared by a previously developed method, genes of which expression level is high are apt to be cloned at a high frequency, whereas genes of which expression level are low, are difficult to be cloned. A low-expression gene has been cloned at very low frequency. Therefore, the gene encoding the key enzyme that is involved in growth disturbance of rice pollen has not been identified. In this study, development of a library preparing method able to correct the expression level was attempted using highly sensitive detection method with radioisotope and some genes related to cold-resistance of rice were isolated. Double strand DNAs were synthesized using mRNA extract from rice anthers and annealed following heat-denaturation. It has been known that single strand DNA molecules abundantly existing in DNA solution can easily aggregate to form double strand DNA, but single stranded DNA molecules poor in the solution are apt to still remain as single strand after annealing. Thus, the amount of single strand DNA would be balanced in the solution between abundant DNA and poor DNA species. The authors succeeded to prepare a gene library including low and high expression genes at similar proportions. Moreover, spin trap method that allows RI labeling of DNA bound to latex particle, was developed to detect with high sensitivity, especially for genes that are expressed at low level. The present method could be used for recovery, detection and quantitative analysis of radiolabeled single strand DNA. Thus, it was demonstrated that the stage from tetrad sperm to small sperm might be easily affected by cold stress. The present results suggest that the expressions of {beta}-1 and {beta}-3 glucanase, which are involved in the release of small sperms following meiosis in the pollen formation, might be easily affected by cold stress. (M.N.)

  1. A new rapid method for isolating nucleoli.

    Science.gov (United States)

    Li, Zhou Fang; Lam, Yun Wah

    2015-01-01

    The nucleolus was one of the first subcellular organelles to be isolated from the cell. The advent of modern proteomic techniques has resulted in the identification of thousands of proteins in this organelle, and live cell imaging technology has allowed the study of the dynamics of these proteins. However, the limitations of current nucleolar isolation methods hinder the further exploration of this structure. In particular, these methods require the use of a large number of cells and tedious procedures. In this chapter we describe a new and improved nucleolar isolation method for cultured adherent cells. In this method cells are snap-frozen before direct sonication and centrifugation onto a sucrose cushion. The nucleoli can be obtained within a time as short as 20 min, and the high yield allows the use of less starting material. As a result, this method can capture rapid biochemical changes in nucleoli by freezing the cells at a precise time, hence faithfully reflecting the protein composition of nucleoli at the specified time point. This protocol will be useful for proteomic studies of dynamic events in the nucleolus and for better understanding of the biology of mammalian cells.

  2. Water feeding method upon reactor isolation

    International Nuclear Information System (INIS)

    Sasaki, Koichi; Takahara, Kuniaki; Hamamura, Kenji; Arakawa, Masahiro.

    1990-01-01

    The present invention concerns a method of feeding water upon reactor isolation in a plural loop type reactor having a plurality of reactor cooling systems. Water can be injected to a plurality of pools even if the pressure between the pools is not balanced and the water level in the reactor cooling system is optimally controlled. That is, water can be injected in accordance with the amount required for each of the pools by setting the opening of a turbine inlet steam control valve to somewhat higher than the cooling system pressure of the highest pressure loop. Water feeding devices upon reactor isolation were required by the same number as that for the reactor cooling systems. Whereas since pumps and turbines are used in common without worsening the water injection controllability to each of the loops according to the method of this invention and, accordingly, the cost performance can be improved. Further, since the opening degree of the turbine inlet steam control valve is controlled while making the difference pressure constant between the turbine inlet pressure and the pump exhaust pressure, the amount of the turbine exhausted steams can be reduced and, further, water injection controllability of the flow rate control valve in the injection line is improved. (I.S.)

  3. Permanent isolation surface barrier development plan

    International Nuclear Information System (INIS)

    Wing, N.R.

    1994-01-01

    The exhumation and treatment of wastes may not always be the preferred alternative in the remediation of a waste site. In-place disposal alternatives, under certain circumstances, may be the most desirable alternatives to use in the protection of human health and the environment. The implementation of an in-place disposal alternative will likely require some type of protective covering that will provide long-term isolation of the wastes from the accessible environment. Even if the wastes are exhumed and treated, a long-term barrier may still be needed to adequately dispose of the treated wastes or any remaining waste residuals. Currently, no open-quotes provenclose quotes long-term barrier is available. The Hanford Site Permanent Isolation Surface Barrier Development Program (BDP) was organized to develop the technology needed to provide a long-term surface barrier capability for the Hanford Site. The permanent isolation barrier technology also could be used at other sites. Permanent isolation barriers use engineered layers of natural materials to create an integrated structure with redundant protective features. Drawings of conceptual permanent isolation surface barriers are shown. The natural construction materials (e.g., fine soil, sand, gravel, riprap, asphalt) have been selected to optimize barrier performance and longevity. The objective of current designs is to use natural materials to develop a maintenance-free permanent isolation surface barrier that isolates wastes for a minimum of 1,000 years by limiting water drainage to near-zero amounts; reducing the likelihood of plant, animal, and human intrusion; controlling the exhalation of noxious gases; and minimizing erosion-related problems

  4. Permanent isolation surface barrier development plan

    Energy Technology Data Exchange (ETDEWEB)

    Wing, N.R.

    1994-01-01

    The exhumation and treatment of wastes may not always be the preferred alternative in the remediation of a waste site. In-place disposal alternatives, under certain circumstances, may be the most desirable alternatives to use in the protection of human health and the environment. The implementation of an in-place disposal alternative will likely require some type of protective covering that will provide long-term isolation of the wastes from the accessible environment. Even if the wastes are exhumed and treated, a long-term barrier may still be needed to adequately dispose of the treated wastes or any remaining waste residuals. Currently, no {open_quotes}proven{close_quotes} long-term barrier is available. The Hanford Site Permanent Isolation Surface Barrier Development Program (BDP) was organized to develop the technology needed to provide a long-term surface barrier capability for the Hanford Site. The permanent isolation barrier technology also could be used at other sites. Permanent isolation barriers use engineered layers of natural materials to create an integrated structure with redundant protective features. Drawings of conceptual permanent isolation surface barriers are shown. The natural construction materials (e.g., fine soil, sand, gravel, riprap, asphalt) have been selected to optimize barrier performance and longevity. The objective of current designs is to use natural materials to develop a maintenance-free permanent isolation surface barrier that isolates wastes for a minimum of 1,000 years by limiting water drainage to near-zero amounts; reducing the likelihood of plant, animal, and human intrusion; controlling the exhalation of noxious gases; and minimizing erosion-related problems.

  5. Development of novel Alicyclobacillus spp. isolation medium.

    Science.gov (United States)

    Chang, S; Kang, D-H

    2005-01-01

    To develop a new isolation medium with higher recovery rates of Alicyclobacillus spp. SK agar was developed with optimized incubation temperature, pH, acidulant, Tween 80 concentration and divalent cation addition. Results indicate that detection of Alicyclobacillus spp. by SK agar was significantly higher (P > 0.05) than those obtained by K agar, orange serum agar, and potato dextrose agar. Current media used for Alicyclobacillus spp. isolation still resulted in high numbers of false negative products. The sensitivity of SK agar to Alicyclobacillus spp. allows detection of low numbers of Alicyclobacillus spp. and also provides a more higher isolation results compared with currently used media. SK agar will be useful to the fruit juice industry to obtain more accurate numbers of contaminant Alicyclobacillus spp. With this media, false negative samples can be reduced, and the likelihood of exported products being rejected can be greatly reduced.

  6. COMPUTER-ASSISTED HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY METHOD DEVELOPMENT WITH APPLICATIONS TO THE ISOLATION AND ANALYSIS OF PHYTOPLANKTON PIGMENTS. (R826944)

    Science.gov (United States)

    We used chromatography modeling software to assist in HPLC method development, with the goalof enhancing separations through the exclusive use of gradient time and column temperature. Wesurveyed nine stationary phases for their utility in pigment purification and natur...

  7. A simple method for DNA isolation from Xanthomonas spp.

    Directory of Open Access Journals (Sweden)

    Gomes Luiz Humberto

    2000-01-01

    Full Text Available A simple DNA isolation method was developed with routine chemicals that yields high quality and integrity preparations when compared to some of the most well known protocols. The method described does not require the use of lysing enzymes, water bath and the DNA was obtained within 40 minutes The amount of nucleic acid extracted (measured in terms of absorbancy at 260 nm from strains of Xanthomonas spp., Pseudomonas spp. and Erwinia spp. was two to five times higher than that of the most commonly used method.

  8. A Modified Method for Isolation of Rhein from Senna

    Science.gov (United States)

    Mehta, Namita; Laddha, K. S.

    2009-01-01

    A simple and efficient method for the isolation of rhein from Cassia angustifolia (senna) leaves is described in which the hydrolysis of the sennosides and extraction of the hydrolysis products (free anthraquinones) is carried out in one step. Further isolation of rhein is achieved from the anthraquinone mixture. This method reduces the number of steps required for isolation of rhein as compared to conventional methods. PMID:20336207

  9. Optimized anion exchange column isolation of zirconium-89 ( 89 Zr) from yttrium cyclotron target: Method development and implementation on an automated fluidic platform

    Energy Technology Data Exchange (ETDEWEB)

    O’Hara, Matthew J.; Murray, Nathaniel J.; Carter, Jennifer C.; Morrison, Samuel S.

    2018-04-01

    Zirconium-89 (89Zr), produced by the (p,n) reaction from naturally monoisotopic yttrium (natY), is a promising positron emitting isotope for immunoPET imaging. Its long half-life of 78.4 h is sufficient for evaluating slow physiological processes. A prototype automated fluidic system, coupled to on-line and in-line detectors, has been constructed to facilitate development of new 89Zr purification methodologies. The highly reproducible reagent delivery platform and near-real time monitoring of column effluents allows for efficient method optimization. The separation of Zr from dissolved Y metal targets was evaluated using several anion exchange resins. Each resin was evaluated against its ability to quantitatively capture Zr from a load solution that is high in dissolved Y. The most appropriate anion exchange resin for this application was identified, and the separation method was optimized. The method is capable of a high Y decontamination factor (>105) and has been shown to separate Fe, an abundant contaminant in Y foils, from the 89Zr elution fraction. Finally, the performance of the method was evaluated using cyclotron bombarded Y foil targets. The separation method was shown to achieve >95% recovery of the 89Zr present in the foils. The 89Zr eluent, however, was in a chemical matrix not immediately conducive to labeling onto proteins. The main intent of this study was to develop a tandem column 89Zr purification process, wherein the anion exchange column method described here is the first separation in a dual-column purification process.

  10. Cooperative method development

    DEFF Research Database (Denmark)

    Dittrich, Yvonne; Rönkkö, Kari; Eriksson, Jeanette

    2008-01-01

    The development of methods tools and process improvements is best to be based on the understanding of the development practice to be supported. Qualitative research has been proposed as a method for understanding the social and cooperative aspects of software development. However, qualitative...... research is not easily combined with the improvement orientation of an engineering discipline. During the last 6 years, we have applied an approach we call `cooperative method development', which combines qualitative social science fieldwork, with problem-oriented method, technique and process improvement....... The action research based approach focusing on shop floor software development practices allows an understanding of how contextual contingencies influence the deployment and applicability of methods, processes and techniques. This article summarizes the experiences and discusses the further development...

  11. An optimized method for mouse liver sinusoidal endothelial cell isolation

    Energy Technology Data Exchange (ETDEWEB)

    Meyer, Jeremy, E-mail: jeremy.meyer@hcuge.ch [Division of Digestive and Transplantation Surgery, University Hospitals of Geneva, Rue Gabrielle-Perret-Gentil 4, 1211 Genève 14 (Switzerland); Unit of Surgical Research, University of Geneva, Rue Michel-Servet 1, 1206 Genève (Switzerland); Lacotte, Stéphanie, E-mail: stephanie.lacotte@unige.ch [Unit of Surgical Research, University of Geneva, Rue Michel-Servet 1, 1206 Genève (Switzerland); Morel, Philippe, E-mail: philippe.morel@hcuge.ch [Division of Digestive and Transplantation Surgery, University Hospitals of Geneva, Rue Gabrielle-Perret-Gentil 4, 1211 Genève 14 (Switzerland); Unit of Surgical Research, University of Geneva, Rue Michel-Servet 1, 1206 Genève (Switzerland); Gonelle-Gispert, Carmen, E-mail: carmen.gonelle@unige.ch [Unit of Surgical Research, University of Geneva, Rue Michel-Servet 1, 1206 Genève (Switzerland); Bühler, Léo, E-mail: leo.buhler@hcuge.ch [Division of Digestive and Transplantation Surgery, University Hospitals of Geneva, Rue Gabrielle-Perret-Gentil 4, 1211 Genève 14 (Switzerland); Unit of Surgical Research, University of Geneva, Rue Michel-Servet 1, 1206 Genève (Switzerland)

    2016-12-10

    The objective of the present study was to develop an accurate and reproducible method for liver sinusoidal endothelial cell (LSEC) isolation in mice. Non-parenchymal cells were isolated using a modified two-step collagenase digestion combined with Optiprep density gradient centrifugation. LSEC were further purified using two prevalent methods, short-term selective adherence and CD146+ magnetic-activated cell sorting (MACS), and compared in terms of cell yield, viability and purity to our purification technique using CD11b cell depletion combined with long-term selective adherence. LSEC purification using our technique allowed to obtain 7.07±3.80 million LSEC per liver, while CD146+ MACS and short-term selective adherence yielded 2.94±1.28 and 0.99±0.66 million LSEC, respectively. Purity of the final cell preparation reached 95.10±2.58% when using our method. In contrast, CD146+ MACS and short-term selective adherence gave purities of 86.75±3.26% and 47.95±9.82%, respectively. Similarly, contamination by non-LSEC was the lowest when purification was performed using our technique, with a proportion of contaminating macrophages of only 1.87±0.77%. Further, isolated cells analysed by scanning electron microscopy presented typical LSEC fenestrations organized in sieve plates, demonstrating that the technique allowed to isolate bona fide LSEC. In conclusion, we described a reliable and reproducible technique for the isolation of high yields of pure LSEC in mice. This protocol provides an efficient method to prepare LSEC for studying their biological functions. - Highlights: • This protocol provides an efficient method to prepare primary mouse LSEC for studying their biological functions. • The liver cell dispersion step was improved by performing a retrograde cannulation of the liver. • The cell yield and the purity obtained were higher than comparative techniques in mice. • Contaminating macrophages were removed by introducing a CD11b- magnetic

  12. An optimized method for mouse liver sinusoidal endothelial cell isolation

    International Nuclear Information System (INIS)

    Meyer, Jeremy; Lacotte, Stéphanie; Morel, Philippe; Gonelle-Gispert, Carmen; Bühler, Léo

    2016-01-01

    The objective of the present study was to develop an accurate and reproducible method for liver sinusoidal endothelial cell (LSEC) isolation in mice. Non-parenchymal cells were isolated using a modified two-step collagenase digestion combined with Optiprep density gradient centrifugation. LSEC were further purified using two prevalent methods, short-term selective adherence and CD146+ magnetic-activated cell sorting (MACS), and compared in terms of cell yield, viability and purity to our purification technique using CD11b cell depletion combined with long-term selective adherence. LSEC purification using our technique allowed to obtain 7.07±3.80 million LSEC per liver, while CD146+ MACS and short-term selective adherence yielded 2.94±1.28 and 0.99±0.66 million LSEC, respectively. Purity of the final cell preparation reached 95.10±2.58% when using our method. In contrast, CD146+ MACS and short-term selective adherence gave purities of 86.75±3.26% and 47.95±9.82%, respectively. Similarly, contamination by non-LSEC was the lowest when purification was performed using our technique, with a proportion of contaminating macrophages of only 1.87±0.77%. Further, isolated cells analysed by scanning electron microscopy presented typical LSEC fenestrations organized in sieve plates, demonstrating that the technique allowed to isolate bona fide LSEC. In conclusion, we described a reliable and reproducible technique for the isolation of high yields of pure LSEC in mice. This protocol provides an efficient method to prepare LSEC for studying their biological functions. - Highlights: • This protocol provides an efficient method to prepare primary mouse LSEC for studying their biological functions. • The liver cell dispersion step was improved by performing a retrograde cannulation of the liver. • The cell yield and the purity obtained were higher than comparative techniques in mice. • Contaminating macrophages were removed by introducing a CD11b- magnetic

  13. Some Orchid Species Fungi Isolated by Different Methods

    Directory of Open Access Journals (Sweden)

    Arzu ÇIĞ

    2014-03-01

    Full Text Available Due to their very small seeds that do not contain endosperm, many terrestrial orchid species require the presence of fungi in order to germinate and maintain their lives; and symbiotic culture studies are being carried out on this topic. For the purpose of determining the orchid species on which the fungus to be used as inoculants in the symbiotic culture will be effective, fungi isolated through several isolation methods are cultured with orchid species. In this study a total of four different isolation methods were applied as one on the tubers and rhizomes and three on the soil of eleven orchid species from the Anacamptis, Cephalanthera, Dactylorhiza and Orchis genera. Three different culture media were used in the methods. At the end of the study Alternaria, Aspergillus, Fusarium, Macrophomina, Rhizoctonia, Trichoderma and Verticillium fungi were isolated. In the study that was conducted with the aimed to isolate particularly Rhizoctania spp. fungi, the fungi was isolated from the tubers of Dactylorhiza umbrosa and Orchis palustris species and the soil of the Orchis simia species. Fusarium and Aspergillus species were isolated the most in all implemented methods and from all species.

  14. GEM simulation methods development

    International Nuclear Information System (INIS)

    Tikhonov, V.; Veenhof, R.

    2002-01-01

    A review of methods used in the simulation of processes in gas electron multipliers (GEMs) and in the accurate calculation of detector characteristics is presented. Such detector characteristics as effective gas gain, transparency, charge collection and losses have been calculated and optimized for a number of GEM geometries and compared with experiment. A method and a new special program for calculations of detector macro-characteristics such as signal response in a real detector readout structure, and spatial and time resolution of detectors have been developed and used for detector optimization. A detailed development of signal induction on readout electrodes and electronics characteristics are included in the new program. A method for the simulation of charging-up effects in GEM detectors is described. All methods show good agreement with experiment

  15. Developments in Surrogating Methods

    Directory of Open Access Journals (Sweden)

    Hans van Dormolen

    2005-11-01

    Full Text Available In this paper, I would like to talk about the developments in surrogating methods for preservation. My main focus will be on the technical aspects of preservation surrogates. This means that I will tell you something about my job as Quality Manager Microfilming for the Netherlands’ national preservation program, Metamorfoze, which is coordinated by the National Library. I am responsible for the quality of the preservation microfilms, which are produced for Metamorfoze. Firstly, I will elaborate on developments in preservation methods in relation to the following subjects: · Preservation microfilms · Scanning of preservation microfilms · Preservation scanning · Computer Output Microfilm. In the closing paragraphs of this paper, I would like to tell you something about the methylene blue test. This is an important test for long-term storage of preservation microfilms. Also, I will give you a brief report on the Cellulose Acetate Microfilm Conference that was held in the British Library in London, May 2005.

  16. A simple method for human peripheral blood monocyte Isolation

    Directory of Open Access Journals (Sweden)

    Marcos C de Almeida

    2000-04-01

    Full Text Available We describe a simple method using percoll gradient for isolation of highly enriched human monocytes. High numbers of fully functional cells are obtained from whole blood or buffy coat cells. The use of simple laboratory equipment and a relatively cheap reagent makes the described method a convenient approach to obtaining human monocytes.

  17. Development of partitioning method

    International Nuclear Information System (INIS)

    Kubota, Kazuo; Dojiri, Shigeru; Kubota, Masumitsu

    1988-10-01

    The literature survey was carried out on the amount of natural resources, behaviors in reprocessing process and in separation and recovery methods of the platinum group elements and technetium which are contained in spent fuel. The essential results are described below. (1) The platinum group elements, which are contained in spent fuel, are quantitatively limited, compared with total demand for them in Japan. And estimated separation and recovery cost is rather high. In spite of that, development of these techniques is considered to be very important because the supply of these elements is almost from foreign resources in Japan. (2) For recovery of these elements, studies of recovery from undisolved residue and from high level liquid waste (HLLW) also seem to be required. (3) As separation and recovery methods, following techniques are considered to be effective; lead extraction, liquid metal extraction, solvent extraction, ion-exchange, adsorption, precipitation, distillation, electrolysis or their combination. (4) But each of these methods has both advantages and disadvantages. So development of such processes largely depends on future works. (author) 94 refs

  18. A rapid and low-cost DNA extraction method for isolating ...

    African Journals Online (AJOL)

    The price of commercial DNA extraction methods makes the routine use of polymerase chain reaction amplification (PCR) based methods rather costly for scientists in developing countries. A guanidium thiocayante-based DNA extraction method was investigated in this study for the isolation of Escherichia coli (E. coli) DNA ...

  19. Evaluation of isolation methods for pathogenic Yersinia enterocolitica from pig intestinal content.

    Science.gov (United States)

    Laukkanen, R; Hakkinen, M; Lundén, J; Fredriksson-Ahomaa, M; Johansson, T; Korkeala, H

    2010-03-01

    The aim of this study was to evaluate the efficiency of four isolation methods for the detection of pathogenic Yersinia enterocolitica from pig intestinal content. The four methods comprised of 15 isolation steps using selective enrichments (irgasan-ticarcillin-potassium chlorate and modified Rappaport broth) and mildly selective enrichments at 4 or 25 degrees C. Salmonella-Shigella-desoxycholate-calcium chloride agar, cefsulodin-irgasan-novobiocin agar were used as plating media. The most sensitive method detected 78% (53/68) of the positive samples. Individual isolation steps using cold enrichment as the only enrichment or as a pre-enrichment step with further selective enrichment showed the highest sensitivities (55-66%). All isolation methods resulted in high numbers of suspected colonies not confirmed as pathogenic Y. enterocolitica. Cold enrichment should be used in the detection of pathogenic Y. enterocolitica from pig intestinal contents. In addition, more than one parallel isolation step is needed. The study shows that depending on the isolation method used for Y. enterocolitica, the detected prevalence of Y. enterocolitica in pig intestinal contents varies greatly. More selective and sensitive isolation methods need to be developed for pathogenic Y. enterocolitica.

  20. An improved method for isolation of RNA from bone

    Directory of Open Access Journals (Sweden)

    Carter Lauren E

    2012-01-01

    Full Text Available Abstract Background Bone physiology is increasingly appreciated as an important contributor to metabolic disorders such as type 2 diabetes. However, progress in understanding the role of bone in determining metabolic health is hampered by the well-described difficulty of obtaining high quality RNA from bone for gene expression analysis using the currently available approaches. Results We developed a simple approach to isolate bone RNA that combines pulverizing the bone and the phenol-guanidinium based RNA extraction in a single step while maintaining near-freezing temperatures. This single step method increases the yield of high quality RNA by eight-fold, with RNA integrity numbers ranging from 6.7 to 9.2. Conclusions Our streamlined approach substantially increases the yield of high-quality RNA from bone tissue while facilitating safe and efficient processing of multiple samples using readily available platforms. The RNA obtained from this method is suitable for use in gene expression analysis in real-time quantitative PCR, microarray, and next generation sequencing applications.

  1. Simple and efficient methods for isolation and activity measurement ...

    African Journals Online (AJOL)

    Jane

    2011-06-29

    Jun 29, 2011 ... Key words: Hirudin, thrombin titration method, chromatography, purification. INTRODUCTION. Since recombinant ... Escherichia coli in 1986, intensive research had been .... mixed with 50 µl sample was incubated in 37°C water for 5 min, then 5 µl .... conclusion, the concise and efficient isolation line of the.

  2. Method for the detection and isolation of traces of organic fluorine compounds in plants

    Energy Technology Data Exchange (ETDEWEB)

    Wade, R H; Ross, J M; Benedict, H M

    1964-01-01

    A method for the detection and isolation of sub-microgram quantities of organic fluorine compounds from plant materials in the presence of much larger amounts of inorganic fluoride is presented. The procedure consists first of a rapid screening step for use with large numbers of vegetable samples and extracts and, second, of a chromatographic step to isolate and characterize any fluoro-organics found. These methods are developed in light of specific chemical characteristics of organic fluorine compounds as a general class. A modification of SOEP's quantitative sub-micro fluoride analytical method is presented as applicable to these isolation methods. Microgram quantities of organic fluorine compounds were found in the plant materials investigated but at a level too low for isolation and identification.

  3. Rapid and inexpensive method for isolating plasmid DNA

    International Nuclear Information System (INIS)

    Aljanabi, S. M.; Al-Awadi, S. J.; Al-Kazaz, A. A.; Baghdad Univ.

    1997-01-01

    A small-scale and economical method for isolating plasmid DNA from bacteria is described. The method provides DNA of suitable quality for most DNA manipulation techniques. This DNA can be used for restriction endonuclease digestion, southern blot hybridization, nick translation and end labeling of DNA probes, Polymerase Chain Reaction (PCR) -based techniques, transformation, DNA cycle-sequencing, and Chain-termination method for DNA sequencing. The entire procedure is adapted to 1.5 ml microfuge tubes and takes approximately 30 mins. The DNA isolated by this method has the same purity produced by CTAB and cesium chloride precipitation and purification procedures respectively. The two previous methods require many hours to obtain the final product and require the use of very expensive equipment as ultracentrifuge. This method is well suited for the isolation of plasmid DNA from a large number of bacterial samples and in a very short time and low cost in laboratories where chemicals, expensive equipment and finance are limited factors in conducting molecular research. (authors). 11refs. 11refs

  4. Comparison of Various Methods for Isolation of Nocardia from Soil

    Directory of Open Access Journals (Sweden)

    Masoumeh Rasouli-Nasab

    2017-02-01

    Full Text Available Background The genus Nocardia is Gram-positive, aerobic filamentous bacilli and saprophytic micro-organisms that can be isolated from freshwater, salt water, dust and decaying vegetation especially the soil. This study aimed to investigate the several media for to determine a suitable culture media with the ability to better for the isolation of Nocardia from soil. Methods In this study, 400 soil samples were collected from different areas from Iran. The soil samples were then cultured on the four culture media such as Humic acid vitamin B agar, Paraffin agar, Sabouraud dextrose agar supplemented whit cycloheximide and carbon-free broth containing paraffin rods and incubated at 35°C. All of culture media investigated every 3 days for a month. Colonies suspicious to Nocardia were stained with Gram-stain, acid-fast and partially acid-fast and evaluated for resistance to lysozyme. Results From 400 soil samples, the number of 62, 10, 28 and 19 strains of Nocardia were isolated by paraffin rods, Humic Acid Vitamin B agar, Paraffin agar and Sabouraud dextrose agar whit cycloheximide, respectively. Most Nocardia strains were isolated using paraffin bait technique. Conclusions Isolation of Nocardia spp. is enhanced by using the paraffin baiting technique that relies on the selective ability of this micro-organism to metabolize paraffin.

  5. Preservation methods for isolates of ascochyta blight fungi

    Directory of Open Access Journals (Sweden)

    Joanna Marcinkowska

    2014-08-01

    Full Text Available Isolates of ascochyta blight fungi, two of Ascochyta pisi, four of Mycosphaerella pinodes and four of Phoma pinodella were stored: A - on slants under mineral oil, B - on CN's medium agar disks, and as conidial suspension: C - in glycerine, D · in water. Viability and pathogenicity of recovered cultures after each consecutive year were assesed from 1991 to 1999. The compared parameters were first of all strongly influenced by the preservation method, but fungus species and number of years had a minor importance. The best for longer storage was method "A" because after 9 years the isolates were viable, highly pathogenic, and cultures recovered from them were clean. Thc method "C'' is good for short keeping (2-3 years, as conidia in vials need only small space and gave clean cultures.

  6. Extraction Methods for the Isolation of Isoflavonoids from Plant Material

    Directory of Open Access Journals (Sweden)

    Blicharski Tomasz

    2017-03-01

    Full Text Available The purpose of this review is to describe and compare selected traditional and modern extraction methods employed in the isolation of isoflavonoids from plants. Conventional methods such as maceration, percolation, or Soxhlet extraction are still frequently used in phytochemical analysis. Despite their flexibility, traditional extraction techniques have significant drawbacks, including the need for a significant investment of time, energy, and starting material, and a requirement for large amounts of potentially toxic solvents. Moreover, these techniques are difficult to automate, produce considerable amount of waste and pose a risk of degradation of thermolabile compounds. Modern extraction methods, such as: ultrasound-assisted extraction, microwave-assisted extraction, accelerated solvent extraction, supercritical fluid extraction, and negative pressure cavitation extraction, can be regarded as remedies for the aforementioned problems. This manuscript discusses the use of the most relevant extraction techniques in the process of isolation of isoflavonoids, secondary metabolites that have been found to have a plethora of biological and pharmacological activities.

  7. Radioactive ion beam production by the ISOL method for SPIRAL

    International Nuclear Information System (INIS)

    Landre-Pellemoine, Frederique

    2001-01-01

    This work is directly related to the SPIRAL project (Systeme de Production d'Ions Radioactifs Acceleres en Lignes) of which the start up will begin in September 2001 at GANIL (Grand Accelerateur National d'Ions Lourds) in Caen. This thesis primarily concerns the development of radioactive ion production systems (target/ion source) by the thorough study of each production stage of the ISOL (Isotopic Separation On Line) method: target and/or projectile fragmentation production, diffusion out of target material, effusion into the ion source and finally the ionization of the radioactive atoms. A bibliographical research and thermal simulations allowed us to optimize materials and the shape of the production and diffusion targets. A first target was optimized and made reliable for the radioactive noble gases production (argon, neon...). A second target dedicated to the radioactive helium production was entirely designed and realised (from the specifications to the 'off line' and 'on line' tests). Finally, a third target source system was defined for singly-charged radioactive alkaline production. The intensities of secondary beams planned for SPIRAL are presented here. A detailed study of the diffusion effusion efficiency for these various targets showed that the use of a fine microstructure carbon (grain size of 1 μm) improved the diffusion and showed the importance of thickness of the lamella for the short lived isotope effusion. (author) [fr

  8. Resource Isolation Method for Program’S Performance on CMP

    Science.gov (United States)

    Guan, Ti; Liu, Chunxiu; Xu, Zheng; Li, Huicong; Ma, Qiang

    2017-10-01

    Data center and cloud computing are more popular, which make more benefits for customers and the providers. However, in data center or clusters, commonly there is more than one program running on one server, but programs may interference with each other. The interference may take a little effect, however, the interference may cause serious drop down of performance. In order to avoid the performance interference problem, the mechanism of isolate resource for different programs is a better choice. In this paper we propose a light cost resource isolation method to improve program’s performance. This method uses Cgroups to set the dedicated CPU and memory resource for a program, aiming at to guarantee the program’s performance. There are three engines to realize this method: Program Monitor Engine top program’s resource usage of CPU and memory and transfer the information to Resource Assignment Engine; Resource Assignment Engine calculates the size of CPU and memory resource should be applied for the program; Cgroups Control Engine divide resource by Linux tool Cgroups, and drag program in control group for execution. The experiment result show that making use of the resource isolation method proposed by our paper, program’s performance can be improved.

  9. Method for strontium isolation from high-mineralized water

    International Nuclear Information System (INIS)

    Evzhanov, Kh.; Andriyasova, G.M.

    1983-01-01

    A method to isolate strontium from high-mineralized waters containing sodium, magnesium, calcium and strontium chlorides, which differ from the prototype method in a considerable decrease in energy consumption with the preservation of a high degree of Sr, Mg and Ca isolation selectivity, has been suggested. According to the method suggested mineralized waters are treated with alkali (NaOH) in the amount of 95-97% of stoichiometry by magnesium, then after separation of magnesium hydroxide precipitate mother liquor is treated with sodium carbonate in the amount of 50-60% of stoichiometry by calcium. After separation of calcium carbonate precipitate mother liquor is treated with NaOH in the amount of 130-135% of stoichiometry by calcium. After separation of calcium hydroxide precipitate from mother liquor by means of sodium carbonate introduction strontium carbonate is isolated. The degree of strontium extraction in the form of SrCO 3 constitutes 90.5% of its content in the initial solution. The method presented can be used for strontium separation from natural and waste waters

  10. A modified efficient method for dental pulp stem cell isolation.

    Science.gov (United States)

    Raoof, Maryam; Yaghoobi, Mohammad Mehdi; Derakhshani, Ali; Kamal-Abadi, Ali Mohammadi; Ebrahimi, Behnam; Abbasnejad, Mehdi; Shokouhinejad, Noushin

    2014-03-01

    Dental pulp stem cells can be used in regenerative endodontic therapy. The aim of this study was to introduce an efficient method for dental pulp stem cells isolation. In this in-vitro study, 60 extracted human third molars were split and pulp tissue was extracted. Dental pulp stem cells were isolated by the following three different methods: (1) digestion of pulp by collagenase/dispase enzyme and culture of the released cells; (2) outgrowth of the cells by culture of undigested pulp pieces; (3) digestion of pulp tissue pieces and fixing them. The cells were cultured in minimum essential medium alpha modification (αMEM) medium supplemented with 20% fetal bovine serum(FBS) in humid 37°C incubator with 5% CO 2. The markers of stem cells were studied by reverse transcriptase polymerase chain reaction (PCR). The student t-test was used for comparing the means of independent groups. P third method, we obtained stem cells successfully with about 60% efficiency after 2 days. The results of RT-PCR suggested the expression of Nanog, Oct-4, and Nucleostemin markers in the isolated cells from dental pulps. This study proposes a new method with high efficacy to obtain dental pulp stem cells in a short time.

  11. An improved method for isolating viruses from asymptomatic carrier fish

    Science.gov (United States)

    Amend, Donald F.; Pietsch, John P.

    1972-01-01

    This paper describes a method using elevated levels of penicillin, streptomycin, and nystatin instead of filters to control bacteria and mold contaminants in specimens processed for virus isolation. Filters were shown to significantly reduce the virus concentration. Virus and tissue cultures were not affected by this procedure. In field tests nearly three times more specimens were positive for virus with this method than with the widely used filter technique. Moreover, the cost of materials was less. This method is recommended for inspection and certification purposes.

  12. A modified efficient method for dental pulp stem cell isolation

    Directory of Open Access Journals (Sweden)

    Maryam Raoof

    2014-01-01

    Full Text Available Background: Dental pulp stem cells can be used in regenerative endodontic therapy. The aim of this study was to introduce an efficient method for dental pulp stem cells isolation. Materials and Methods: In this in-vitro study, 60 extracted human third molars were split and pulp tissue was extracted. Dental pulp stem cells were isolated by the following three different methods: (1 digestion of pulp by collagenase/dispase enzyme and culture of the released cells; (2 outgrowth of the cells by culture of undigested pulp pieces; (3 digestion of pulp tissue pieces and fixing them. The cells were cultured in minimum essential medium alpha modification (αMEM medium supplemented with 20% fetal bovine serum(FBS in humid 37°C incubator with 5% CO 2 . The markers of stem cells were studied by reverse transcriptase polymerase chain reaction (PCR. The student t-test was used for comparing the means of independent groups. P <0.05 was considered as significant. Results: The results indicated that by the first method a few cell colonies with homogenous morphology were detectable after 4 days, while in the outgrowth method more time was needed (10-12 days to allow sufficient numbers of heterogeneous phenotype stem cells to migrate out of tissue. Interestingly, with the improved third method, we obtained stem cells successfully with about 60% efficiency after 2 days. The results of RT-PCR suggested the expression of Nanog, Oct-4, and Nucleostemin markers in the isolated cells from dental pulps. Conclusion: This study proposes a new method with high efficacy to obtain dental pulp stem cells in a short time.

  13. A molecular method for typing Herpes simplex virus isolates as an alternative to immunofluorescence methods

    Directory of Open Access Journals (Sweden)

    Abraham A

    2009-01-01

    Full Text Available Background: Typing of Herpes simplex virus (HSV isolates is required to identify the virus isolated in culture. The methods available for this include antigen detection by immunofluorescence (IF assays and polymerase chain reaction (PCR. This study was undertaken to standardize a molecular method for typing of HSV and compare it with a commercial IF reagent for typing. Objectives: To compare a molecular method for typing HSV isolates with a monoclonal antibody (MAb based IF test. Study design : This cross-sectional study utilized four reference strains and 42 HSV isolates obtained from patients between September 1998 and September 2004. These were subjected to testing using an MAb-based IF test and a PCR that detects the polymerase ( pol gene of HSV isolates. Results: The observed agreement of the MAb IF assay with the pol PCR was 95.7%. Fifty four point eight percent (23/42 of isolates tested by IF typing were found to be HSV-1, 40.5% (17/42 were HSV-2, and two (4.8% were untypable using the MAb IF assay. The two untypable isolates were found to be HSV-2 using the pol PCR. In addition, the cost per PCR test for typing is estimated to be around Rs 1,300 (USD 30, whereas the cost per MAb IF test is about Rs 1,500 (USD 35 including all overheads (reagents, instruments, personnel time, and consumables. Conclusion: The pol PCR is a cheaper and more easily reproducible method for typing HSV isolates as compared to the IF test. It could replace the IF-based method for routine typing of HSV isolates as availability of PCR machines (thermal cyclers is now more widespread than fluorescence microscopes in a country like India.

  14. Participation in sizing methods and management improvement for the isolated hybrid systems coupling photovoltaic generator and power generating unit. Application in developing countries; Participation a l'amelioration des methodes de dimensionnement et de gestion pour les systemes isoles hybrides de production d'electricite, couplant generateur photovoltaique et groupe electrogene. Application dans les pays en voie de developpement

    Energy Technology Data Exchange (ETDEWEB)

    Camez, C.

    2004-09-15

    One of the applications justifying more the use of renewable energies is the electrification of isolated sites. Sites for which connection to the local distribution network is being out of price or impossible. These sites are most of the time electrified with a diesel generator. This technology is tested and mature however two major disadvantages can be raised here: necessary routine maintenance and the fuel provisioning management. Photovoltaic system can answer here the request and little solar home systems (100 Watts) are installed. In the particular situation of a village electrification, the consumers quickly see in these systems a strong limitation with the development. Reliability, evolution, comfort, service are the masters words here, the development of a micro industry also seems necessary to a village development. It is then necessary to propose a micro network type solution. Such a generator cannot be think completely photovoltaic, presenting an important over-cost compared to a conventional solution. The idea of conventional and renewable energy coupling is born. Such a multi-source system: hybrid, implies an increased sizing and management complexity. An analysis software tool was developed: HYPSIM, this one makes it possible to optimize the operation of such a system. HYPSIM made possible to - Justify the relevance of a photovoltaic/diesel generator/batteries system. - Conclude on the sizing criteria to be observed. (author)

  15. Development of partitioning method

    International Nuclear Information System (INIS)

    Kondo, Yasuo; Kubota, Masumitsu; Abe, Tadashi; Nagato, Kotaro.

    1991-09-01

    Spent fuels from nuclear power stations contain many useful elements, which can be utilized as heat and irradiation sources, radioisotope, elemental resource, etc. Their recovery from spent fuel and effective uses have the advantages in not only converting the radioactive waste to beneficial resources but also promoting rationalization of the management and disposal of the radioactive wastes. In present study, published literature related to recovery and utilization of useful elements in spent fuel was mainly surveyed, present states and trends in their research and development were analyzed, and their future prospects were conjectured. Research and development on recovery and utilization of useful elements are being continued mainly in USA, Europe and Japan. A transportable food irradiator with Cs-137 and an electric power source with Sr-90 for remote weather station are typical examples in major past applications. However, research and development on recovery and utilization are not so much active and the future efforts should be expected hereafter. Present study was conducted under the auspices of the Science and Technology Agency of Japan. (author)

  16. Development of partitioning method

    International Nuclear Information System (INIS)

    Kobayashi, Tsutomu; Shirahashi, Koichi; Kubota, Masumitsu

    1989-11-01

    Precipitation behavior of elements in a high-level liquid waste (HLW) was studied by using the simulated liquid waste, when the transuranic elements group was precipitated and separated as oxalate from HLW generated from the reprocessing of spent nuclear fuel. The results showed that over 90 % of strontium and barium were precipitated when oxalic acid was directly added to HLW to precipitate the transuranic elements group, and the percentages of these elements precipitated were affected by molybdenum and or zirconium. Therefore, a method of adding oxalic acid into the filtrate was studied after removing previously molybdenum and zirconium as precipitate by denitrating HLW, and it was found that precipitated fractions of strontium and barium could be suppressed about 10 %. Adding oxalic acid under the co-existance of ascorbic acid is effective for quantitative precipitation of neptunium in HLW. In this case, it was found that adding ascorbic acid had little influence on precipitation behavior of the other elements except palladium. (author)

  17. A safe inexpensive method to isolate high quality plant and fungal ...

    African Journals Online (AJOL)

    The most commonly used plant DNA isolation methods use toxic and hazardous chemicals (phenol, chloroform), which require special equipment to minimize exposure and may limit their use in certain environments. Commercial DNA extraction kits are convenient and usually safe, but their availability to certain developing ...

  18. A method for the isolation of root hairs from the model legume Medicago truncatula

    NARCIS (Netherlands)

    Ramos Escribano, J.; Bisseling, T.

    2003-01-01

    A new method for the isolation of root hairs from the model legume, Medicago truncatula, was developed. The procedure involves the propagation of detached roots on agar plates and the collection of root hairs by immersion in liquid nitrogen. Yields of up to 40 µg of root hair protein were obtained

  19. Deep sequencing as a method of typing bluetongue virus isolates.

    Science.gov (United States)

    Rao, Pavuluri Panduranga; Reddy, Yella Narasimha; Ganesh, Kapila; Nair, Shreeja G; Niranjan, Vidya; Hegde, Nagendra R

    2013-11-01

    Bluetongue (BT) is an economically important endemic disease of livestock in tropics and subtropics. In addition, its recent spread to temperate regions like North America and Northern Europe is of serious concern. Rapid serotyping and characterization of BT virus (BTV) is an essential step in the identification of origin of the virus and for controlling the disease. Serotyping of BTV is typically performed by serum neutralization, and of late by nucleotide sequencing. This report describes the near complete genome sequencing and typing of two isolates of BTV using Illumina next generation sequencing platform. Two of the BTV RNAs were multiplexed with ten other unknown samples. Viral RNA was isolated and fragmented, reverse transcribed, the cDNA ends were repaired and ligated with a multiplex oligo. The genome library was amplified using primers complementary to the ligated oligo and subjected to single and paired end sequencing. The raw reads were assembled using a de novo method and reference-based assembly was performed based on the contig data. Near complete sequences of all segments of BTV were obtained with more than 20× coverage, and single read sequencing method was sufficient to identify the genotype and serotype of the virus. The two viruses used in this study were typed as BTV-1 and BTV-9E. Copyright © 2013 Elsevier B.V. All rights reserved.

  20. Development of thermoelectric generators for electrification of isolated rural homes

    Energy Technology Data Exchange (ETDEWEB)

    Rinalde, G.F.; Taglialavore, E.; Gortari, S. [CNEA (National Atomic Energy Commission), Centro Atomico Bariloche, 8400 Bariloche (Argentina); Juanico, L.E. [Conicet (National Scientific and Technologic Research Council), Centro Atomico Bariloche, 8400 Bariloche (Argentina); Molina, M.G. [CONICET and Universidad Nacional de San Juan, Av. Libertador San Martin Oeste, 1109, 5400, San Juan (Argentina)

    2010-06-15

    This work presents the experimental development of the first two prototypes of thermoelectric generators intended for initial electrification of rural isolated homes. The microcontroller system designed for these devices is oriented to develop a ''plug and play'' generator that is able to work on firewood home stoves without specialized supervision. (author)

  1. ADVANCED SEISMIC BASE ISOLATION METHODS FOR MODULAR REACTORS

    Energy Technology Data Exchange (ETDEWEB)

    E. Blanford; E. Keldrauk; M. Laufer; M. Mieler; J. Wei; B. Stojadinovic; P.F. Peterson

    2010-09-20

    Advanced technologies for structural design and construction have the potential for major impact not only on nuclear power plant construction time and cost, but also on the design process and on the safety, security and reliability of next generation of nuclear power plants. In future Generation IV (Gen IV) reactors, structural and seismic design should be much more closely integrated with the design of nuclear and industrial safety systems, physical security systems, and international safeguards systems. Overall reliability will be increased, through the use of replaceable and modular equipment, and through design to facilitate on-line monitoring, in-service inspection, maintenance, replacement, and decommissioning. Economics will also receive high design priority, through integrated engineering efforts to optimize building arrangements to minimize building heights and footprints. Finally, the licensing approach will be transformed by becoming increasingly performance based and technology neutral, using best-estimate simulation methods with uncertainty and margin quantification. In this context, two structural engineering technologies, seismic base isolation and modular steel-plate/concrete composite structural walls, are investigated. These technologies have major potential to (1) enable standardized reactor designs to be deployed across a wider range of sites, (2) reduce the impact of uncertainties related to site-specific seismic conditions, and (3) alleviate reactor equipment qualification requirements. For Gen IV reactors the potential for deliberate crashes of large aircraft must also be considered in design. This report concludes that base-isolated structures should be decoupled from the reactor external event exclusion system. As an example, a scoping analysis is performed for a rectangular, decoupled external event shell designed as a grillage. This report also reviews modular construction technology, particularly steel-plate/concrete construction using

  2. ADVANCED SEISMIC BASE ISOLATION METHODS FOR MODULAR REACTORS

    International Nuclear Information System (INIS)

    Blanford, E.; Keldrauk, E.; Laufer, M.; Mieler, M.; Wei, J.; Stojadinovic, B.; Peterson, P.F.

    2010-01-01

    Advanced technologies for structural design and construction have the potential for major impact not only on nuclear power plant construction time and cost, but also on the design process and on the safety, security and reliability of next generation of nuclear power plants. In future Generation IV (Gen IV) reactors, structural and seismic design should be much more closely integrated with the design of nuclear and industrial safety systems, physical security systems, and international safeguards systems. Overall reliability will be increased, through the use of replaceable and modular equipment, and through design to facilitate on-line monitoring, in-service inspection, maintenance, replacement, and decommissioning. Economics will also receive high design priority, through integrated engineering efforts to optimize building arrangements to minimize building heights and footprints. Finally, the licensing approach will be transformed by becoming increasingly performance based and technology neutral, using best-estimate simulation methods with uncertainty and margin quantification. In this context, two structural engineering technologies, seismic base isolation and modular steel-plate/concrete composite structural walls, are investigated. These technologies have major potential to (1) enable standardized reactor designs to be deployed across a wider range of sites, (2) reduce the impact of uncertainties related to site-specific seismic conditions, and (3) alleviate reactor equipment qualification requirements. For Gen IV reactors the potential for deliberate crashes of large aircraft must also be considered in design. This report concludes that base-isolated structures should be decoupled from the reactor external event exclusion system. As an example, a scoping analysis is performed for a rectangular, decoupled external event shell designed as a grillage. This report also reviews modular construction technology, particularly steel-plate/concrete construction using

  3. A simple and efficient method for isolating small RNAs from different plant species

    Directory of Open Access Journals (Sweden)

    de Folter Stefan

    2011-02-01

    Full Text Available Abstract Background Small RNAs emerged over the last decade as key regulators in diverse biological processes in eukaryotic organisms. To identify and study small RNAs, good and efficient protocols are necessary to isolate them, which sometimes may be challenging due to the composition of specific tissues of certain plant species. Here we describe a simple and efficient method to isolate small RNAs from different plant species. Results We developed a simple and efficient method to isolate small RNAs from different plant species by first comparing different total RNA extraction protocols, followed by streamlining the best one, finally resulting in a small RNA extraction method that has no need of first total RNA extraction and is not based on the commercially available TRIzol® Reagent or columns. This small RNA extraction method not only works well for plant tissues with high polysaccharide content, like cactus, agave, banana, and tomato, but also for plant species like Arabidopsis or tobacco. Furthermore, the obtained small RNA samples were successfully used in northern blot assays. Conclusion Here we provide a simple and efficient method to isolate small RNAs from different plant species, such as cactus, agave, banana, tomato, Arabidopsis, and tobacco, and the small RNAs from this simplified and low cost method is suitable for downstream handling like northern blot assays.

  4. Development of a robust method for isolation of shiga toxin-positive Escherichia coli (STEC from fecal, plant, soil and water samples from a leafy greens production region in California.

    Directory of Open Access Journals (Sweden)

    Michael B Cooley

    Full Text Available During a 2.5-year survey of 33 farms and ranches in a major leafy greens production region in California, 13,650 produce, soil, livestock, wildlife, and water samples were tested for Shiga toxin (stx-producing Escherichia coli (STEC. Overall, 357 and 1,912 samples were positive for E. coli O157:H7 (2.6% or non-O157 STEC (14.0%, respectively. Isolates differentiated by O-typing ELISA and multilocus variable number tandem repeat analysis (MLVA resulted in 697 O157:H7 and 3,256 non-O157 STEC isolates saved for further analysis. Cattle (7.1%, feral swine (4.7%, sediment (4.4%, and water (3.3% samples were positive for E. coli O157:H7; 7/32 birds, 2/145 coyotes, 3/88 samples from elk also were positive. Non-O157 STEC were at approximately 5-fold higher incidence compared to O157 STEC: cattle (37.9%, feral swine (21.4%, birds (2.4%, small mammals (3.5%, deer or elk (8.3%, water (14.0%, sediment (12.3%, produce (0.3% and soil adjacent to produce (0.6%. stx1, stx2 and stx1/stx2 genes were detected in 63%, 74% and 35% of STEC isolates, respectively. Subtilase, intimin and hemolysin genes were present in 28%, 25% and 79% of non-O157 STEC, respectively; 23% were of the "Top 6″ O-types. The initial method was modified twice during the study revealing evidence of culture bias based on differences in virulence and O-antigen profiles. MLVA typing revealed a diverse collection of O157 and non-O157 STEC strains isolated from multiple locations and sources and O157 STEC strains matching outbreak strains. These results emphasize the importance of multiple approaches for isolation of non-O157 STEC, that livestock and wildlife are common sources of potentially virulent STEC, and evidence of STEC persistence and movement in a leafy greens production environment.

  5. Method Development in Forensic Toxicology.

    Science.gov (United States)

    Peters, Frank T; Wissenbach, Dirk K; Busardo, Francesco Paolo; Marchei, Emilia; Pichini, Simona

    2017-01-01

    In the field of forensic toxicology, the quality of analytical methods is of great importance to ensure the reliability of results and to avoid unjustified legal consequences. A key to high quality analytical methods is a thorough method development. The presented article will provide an overview on the process of developing methods for forensic applications. This includes the definition of the method's purpose (e.g. qualitative vs quantitative) and the analytes to be included, choosing an appropriate sample matrix, setting up separation and detection systems as well as establishing a versatile sample preparation. Method development is concluded by an optimization process after which the new method is subject to method validation. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  6. A combined dynamic analysis method for geometrically nonlinear vibration isolators with elastic rings

    Science.gov (United States)

    Hu, Zhan; Zheng, Gangtie

    2016-08-01

    A combined analysis method is developed in the present paper for studying the dynamic properties of a type of geometrically nonlinear vibration isolator, which is composed of push-pull configuration rings. This method combines the geometrically nonlinear theory of curved beams and the Harmonic Balance Method to overcome the difficulty in calculating the vibration and vibration transmissibility under large deformations of the ring structure. Using the proposed method, nonlinear dynamic behaviors of this isolator, such as the lock situation due to the coulomb damping and the usual jump resulting from the nonlinear stiffness, can be investigated. Numerical solutions based on the primary harmonic balance are first verified by direct integration results. Then, the whole procedure of this combined analysis method is demonstrated and validated by slowly sinusoidal sweeping experiments with different amplitudes of the base excitation. Both numerical and experimental results indicate that this type of isolator behaves as a hardening spring with increasing amplitude of the base excitation, which makes it suitable for isolating both steady-state vibrations and transient shocks.

  7. Social isolation and brain development in the ant Camponotus floridanus.

    Science.gov (United States)

    Seid, Marc A; Junge, Erich

    2016-06-01

    Social interactions play a key role in the healthy development of social animals and are most pronounced in species with complex social networks. When developing offspring do not receive proper social interaction, they show developmental impairments. This effect is well documented in mammalian species but controversial in social insects. It has been hypothesized that the enlargement of the mushroom bodies, responsible for learning and memory, observed in social insects is needed for maintaining the large social networks and/or task allocation. This study examines the impact of social isolation on the development of mushroom bodies of the ant Camponotus floridanus. Ants raised in isolation were shown to exhibit impairment in the growth of the mushroom bodies as well as behavioral differences when compared to ants raised in social groups. These results indicate that social interaction is necessary for the proper development of C. floridanus mushroom bodies.

  8. Social isolation and brain development in the ant Camponotus floridanus

    Science.gov (United States)

    Seid, Marc A.; Junge, Erich

    2016-06-01

    Social interactions play a key role in the healthy development of social animals and are most pronounced in species with complex social networks. When developing offspring do not receive proper social interaction, they show developmental impairments. This effect is well documented in mammalian species but controversial in social insects. It has been hypothesized that the enlargement of the mushroom bodies, responsible for learning and memory, observed in social insects is needed for maintaining the large social networks and/or task allocation. This study examines the impact of social isolation on the development of mushroom bodies of the ant Camponotus floridanus. Ants raised in isolation were shown to exhibit impairment in the growth of the mushroom bodies as well as behavioral differences when compared to ants raised in social groups. These results indicate that social interaction is necessary for the proper development of C. floridanus mushroom bodies.

  9. Isolated Liver Perfusion Using Percutaneous Methods:[ql An Experimental Study in the Pig

    International Nuclear Information System (INIS)

    Harnek, Jan; Cwikiel, Wojciech; Bergqvist, Lennart; Persson, Bo; Stridbeck, Hans

    1996-01-01

    Purpose: To develop a method for isolated perfusion of the liver using radiological methods. Methods: Twenty-one pigs, weighing about 20 kg, were divided into three groups. By transjugular and transfemoral approaches two occlusion balloons were placed in the inferior vena cava cranial and caudal, respectively, to the origin of the hepatic veins. One occlusion balloon was placed transfemorally in the common hepatic artery. Another occlusion balloon was inserted in the main branch of the portal vein via the transjugular-transhepatic approach in 11 pigs (groups 1 and 2), and in 10 pigs (group 3) by a percutaneous transhepatic route. After inflation of the balloons, patency of the isolated liver circulation was evaluated by recirculation of 99 Tc m -labelled human albumin during 30 min. Blood tests were obtained after 1, 3, 5, 10, 15, and 30 min to evaluate leakage from the liver to the systemic circulation. Results: Increasing leakage to the systemic circulation from the isolated liver circulation was observed in groups 1 and 2. In the third group the leakage was less than 10%. Conclusion: In an experimental animal model, isolated perfusion of the liver with minor leakage to the systemic circulation may be achieved using radiological methods

  10. Total error components - isolation of laboratory variation from method performance

    International Nuclear Information System (INIS)

    Bottrell, D.; Bleyler, R.; Fisk, J.; Hiatt, M.

    1992-01-01

    The consideration of total error across sampling and analytical components of environmental measurements is relatively recent. The U.S. Environmental Protection Agency (EPA), through the Contract Laboratory Program (CLP), provides complete analyses and documented reports on approximately 70,000 samples per year. The quality assurance (QA) functions of the CLP procedures provide an ideal data base-CLP Automated Results Data Base (CARD)-to evaluate program performance relative to quality control (QC) criteria and to evaluate the analysis of blind samples. Repetitive analyses of blind samples within each participating laboratory provide a mechanism to separate laboratory and method performance. Isolation of error sources is necessary to identify effective options to establish performance expectations, and to improve procedures. In addition, optimized method performance is necessary to identify significant effects that result from the selection among alternative procedures in the data collection process (e.g., sampling device, storage container, mode of sample transit, etc.). This information is necessary to evaluate data quality; to understand overall quality; and to provide appropriate, cost-effective information required to support a specific decision

  11. The development of base-isolated APWR plants

    International Nuclear Information System (INIS)

    Tanaka, T.; Nitta, T.

    2001-01-01

    The full text follows: The seismic design of nuclear power stations plays a critical role in the assurance of plant safety in Japan, and standardization of design is difficult to achieve because every site is subject to different seismic conditions. However, the introduction of seismic -isolation devices is one way to rationally achieve safety assurance and promote design standardization. Base-isolated APWR (advanced pressurized water reactor) plants were developed by applying seismic -isolation devices to APWR plants. The introduction of seismic -isolation devices, which are installed between the ground and buildings, largely decreases the effect of seismic force on buildings. Therefore, the limitation of building shape and eccentricity, which are undertaken in order to prevent the floating of buildings, could be eliminated. This permits the flexibility of building layouts, which result in a reduction of building volume. At the same time, the thickness of the buildings walls that are specific to nuclear power stations, can also be decreased except radiation shield. As for the base-isolated APWR equipment design, the rational design of support structures for equipment and pipings is possible, because the floor response acceleration is greatly reduced. For the cost reduction, it has been confirmed that the base-isolated APWR plants are more economical than traditional APWR plants even after the additionally required expenses for seismic-isolation devices are taken into account. This is primarily because of the rational design of the buildings and equipment which is possible as described above. Another advantage is that building standardization can be promoted because the seismic-isolation devices are able to control the seismic force transmitted to the buildings. This is accomplished by arranging the characteristics of the isolation devices according to the seismic conditions of each site. The introduction of these devices to nuclear power stations is nearly ready

  12. A 3-D aerodynamic method for the analysis of isolated horizontal-axis wind turbines

    Energy Technology Data Exchange (ETDEWEB)

    Ammara, I.; Masson, C.; Paraschivoiu, I. [Ecole Polytechnique, Montreal (Canada)

    1997-12-31

    In most existing performance-analysis methods, wind turbines are considered isolated so that interference effects caused by other rotors or by the site topography are neglected. The main objective of this paper is to propose a practical 3-D method suitable for the study of these effects, in order to optimize the arrangement and the positioning of Horizontal-Axis Wind Turbines (HAWTs) in a wind farm. In the proposed methodology, the flow field around isolated HAWTs is predicted by solving the 3-D, time-averaged, steady-state, incompressible, Navier-Stokes equations in which the turbines are represented by distributions of momentum sources. The resulting governing equations are solved using a Control-Volume Finite Element Method (CVFEM). The fundamental aspects related to the development of a practical 3-D method are discussed in this paper, with an emphasis on some of the challenges that arose during its implementation. The current implementation is limited to the analysis of isolated HAWTs. Preliminary results have indicated that, the proposed 3-D method reaches the same level of accuracy, in terms of performance predictions, that the previously developed 2-D axisymmetric model and the well-known momentum-strip theory, while still using reasonable computers resources. It can be considered as a useful tool for the design of HAWTs. Its main advantages, however, are its intrinsic capacity to predict the details of the flow in the wake, and its capabilities of modelling arbitrary wind-turbine arrangements and including ground effects.

  13. Development of a 3-dimensional seismic isolation floor for computer systems

    International Nuclear Information System (INIS)

    Kurihara, M.; Shigeta, M.; Nino, T.; Matsuki, T.

    1991-01-01

    In this paper, we investigated the applicability of a seismic isolation floor as a method for protecting computer systems from strong earthquakes, such as computer systems in nuclear power plants. Assuming that the computer system is guaranteed for 250 cm/s 2 of input acceleration in the horizontal and vertical directions as the seismic performance, the basic design specification of the seismic isolation floor is considered as follows. Against S 1 level earthquakes, the maximum acceleration response of the seismic isolation floor in the horizontal and vertical directions is kept less than 250 cm/s 2 to maintain continuous computer operation. Against S 2 level earthquakes, the isolation floor allows large horizontal movement and large displacement of the isolation devices to reduce the acceleration response, although it is not guaranteed to be less than 250 cm/s 2 . By reducing the acceleration response, however, serious damage to the computer systems is reduced, so that they can be restarted after an earthquake. Usually, seismic isolation floor systems permit 2-dimensional (horizontal) isolation. However, in the case of just-under-seated earthquakes, which have large vertical components, the vertical acceleration response of this system is amplified by the lateral vibration of the frame of the isolation floor. Therefore, in this study a 3-dimensional seismic isolation floor, including vertical isolation, was developed. This paper describes 1) the experimental results of the response characteristics of the 3-dimensional seismic isolation floor built as a trial using a 3-dimensional shaking table, and 2) comparison of a 2-dimensional analytical model, for motion in one horizontal direction and the vertical direction, to experimental results. (J.P.N.)

  14. Biological Methods and Manual Development

    Science.gov (United States)

    EPA scientists conduct research to develop and evaluate analytical methods for the identification, enumeration, evaluation of aquatic organisms exposed to environmental stressors and to correlate exposures with effects on chemical and biological indicators

  15. DDOT MXD+ method development report.

    Science.gov (United States)

    2015-09-01

    Mixed-use development has become increasingly common across the country, including Washington, D.C. : However, a straightforward and empirically validated method for evaluating the traffic impacts of such : projects is still needed. The data presente...

  16. Negotiating a Systems Development Method

    Science.gov (United States)

    Karlsson, Fredrik; Hedström, Karin

    Systems development methods (or methods) are often applied in tailored version to fit the actual situation. Method tailoring is in most the existing literature viewed as either (a) a highly rational process with the method engineer as the driver where the project members are passive information providers or (b) an unstructured process where the systems developer makes individual choices, a selection process without any driver. The purpose of this chapter is to illustrate that important design decisions during method tailoring are made by project members through negotiation. The study has been carried out using the perspective of actor-network theory. Our narratives depict method tailoring as more complex than (a) and (b) show the driver role rotates between the project members, and design decisions are based on influences from several project members. However, these design decisions are not consensus decisions.

  17. Proglobulin processing enzyme in vacuoles isolated from developing pumpkin cotyledons

    International Nuclear Information System (INIS)

    Hara-Nishimura, I.; Nishimura, M.

    1987-01-01

    The enzymic conversion of proglobulin to globulin catalyzed by the extracts of vacuoles isolated from developing pumpkin (Cucurbita sp. cv Kurokawa Amakuri Nankin) cotyledons was investigated. The endoplasmic reticulum fraction isolated from the developing cotyledons pulse-labeled with [ 35 S]methionine was shown to contain mainly the radiolabeled proglobulin, which was used as a substrate for assaying the proteolytic processing in vitro. The vacuolar extracts catalyzed the proteolytic processing of the proglobulin molecule to produce globulin containing two kinds of polypeptide chains, γ and δ. The pH optimum for the vacuole-mediated conversion was at pH 5.0. The proteolytic processing of proglobulin by the vacuolar extracts was inhibited in the presence of various thiol reagents, e.g. p-chloromercuribenzoate, N-ethylmaleimide, iodoacetic acid, Hg 2+ , and Cu 2+ , but not phenylmethylsulfonyl fluoride, EDTA, o-phenanthroline, leupeptin, antipain, pepstatin, chymostatin, or pumpkin trypsin inhibitor, and was activated in the presence of dithiothreitol and cysteine, indicating that the processing enzyme is a thiol protease. The suborganellar fractionation of the vacuoles showed that the processing activity was localized in the matrix fraction, but not in the membrane or crystalloid fractions. During the seed development, the enzyme was shown to increase, exhibiting the maximal activity at the late developmental stage. The matrix fraction of the protein bodies isolated from the dry castor bean (Ricinus communis) exhibited the processing activity toward the pumpkin proglobulin molecules in the same manner as that by the matrix fraction of pumpkin vacuoles

  18. Development of Probabilistic Performance Evaluation Procedure for Umbilical Lines of Seismically Isolated NPPs

    International Nuclear Information System (INIS)

    Hahm, Daegi; Park, Junhee; Choi, Inkil

    2013-01-01

    In this study, we proposed a procedure to perform the probabilistic performance evaluation of interface piping system for seismically isolated NPPs, and carried out the preliminary performance evaluation of the target example umbilical line. For EDB level earthquakes, the target performance goal cannot be fulfilled, but we also find out that the result can be changed with respect to the variation of the assumed values, i. e., the distribution of response, and the limit state of piping system. Recently, to design the nuclear power plants (NPPs) more efficiently and safely against the strong seismic load, many researchers focus on the seismic isolation system. For the adoption of seismic isolation system to the NPPs, the seismic performance of isolation devices, structures, and components should be guaranteed firstly. Hence, some researches were performed to determine the seismic performance of such items. For the interface piping system between isolated structure and non-isolated structure, the seismic capacity should be carefully estimated since that the required displacement absorption capacity will be increased significantly by the adoption of the seismic isolation system. Nowadays, in NUREG report, the probabilistic performance criteria for isolated NPP structures and components are proposed. Hence, in this study, we developed the probabilistic performance evaluation method and procedure for interface piping system, and applied the method to an example pipe. The detailed procedure and main results are summarized in next section. For the interface piping system, the seismic capacity should be carefully estimated since that the required displacement absorption capacity will be increased significantly by the adoption of the seismic isolation system

  19. Comparative miRNA Analysis of Urine Extracellular Vesicles Isolated through Five Different Methods

    Directory of Open Access Journals (Sweden)

    Felix Royo

    2016-12-01

    Full Text Available Urine extracellular vesicles are a valuable low-invasive source of information, especially for the cells of the genitourinary tract. In the search for biomarkers, different techniques have been developed to isolate and characterize the cargo of these vesicles. In the present work, we compare five of these different isolation methods (three commercial isolation kits, ultracentrifugation, and lectin-based purification and perform miRNA profiling using a multiplex miRNA assay. The results showed high correlation through all isolation techniques, and 48 out of 68 miRNAs were detected above the detection limit at least 10 times. The results obtained by multiplex assay were validated through Taqman qPCR. In addition, using this technique combined with a clinically friendly extracellular vesicle (uEV-enrichment method, we performed the analysis of selected miRNAs in urine from patients affected with bladder cancer, benign prostate hyperplasia, or prostate cancer. Importantly, we found that those miRNAs could be detected in almost 100% of the samples, and no significant differences were observed between groups. Our results support the feasibility of analyzing exosomes-associated miRNAs using a methodology that requires a small volume of urine and is compatible with a clinical environment and high-throughput analysis.

  20. Laser Ion Source Development for ISOL Systems at RIA

    CERN Document Server

    Liu, Yuan; Beene, James R; Bilheux, Hassina Z; Brueck, Kim; Geppert, Christopher; Havener, Charles; Kessler, Thomas; Krause, Herbert F; Schultz, David R; Stracener, Dan; Vane, C R; Wendt, Klaus

    2005-01-01

    The isobaric purity of radioactive ion beams (RIBs) is of crucial importance to many experiments. Laser ion sources based on resonant photoionization have already proved to be of great value at existing ISOL RIB facilities. In these ion sources, ions of a selected isotope are produced by laser radiation via stepwise atomic resonant excitations followed by ionization in the last transition. Because each element has its own unique atomic energy levels, the resonant photoionization process can provide elemental selectivity of nearly 100%. We have initiated a research effort to develop a prototype laser ion source with the potential to achieve the high selectivity and high efficiency required for research with ISOL-generated RIBs at the Rare Isotope Accelerator (RIA). A pilot experiment has been conducted to demonstrate resonant photoionization of three atomic species using all-solid-state tunable Ti:Sapphire lasers. Three Ti:Sapphire lasers were provided by the University of Mainz and used in the experiment for ...

  1. A fast method for large-scale isolation of phages from hospital ...

    African Journals Online (AJOL)

    This plaque-forming method could be adopted to isolate E. coli phage easily, rapidly and in large quantities. Among the 18 isolated E. coli phages, 10 of them had a broad host range in E. coli and warrant further study. Key words: Escherichia coli phages, large-scale isolation, drug resistance, biological properties.

  2. Development of Seismic Isolation Systems Using Periodic Materials

    Energy Technology Data Exchange (ETDEWEB)

    Yan, Yiqun [Univ. of Houston, Houston, TX (United States); Mo, Yi-Lung [Univ. of Houston, Houston, TX (United States); Menq, Farn-Yuh [Univ. of Texas, Austin, TX (United States); Stokoe, II, Kenneth H. [Univ. of Texas, Austin, TX (United States); Perkins, Judy [Prairie View A & M University, Prairie View, TX (United States); Tang, Yu [Argonne National Lab. (ANL), Argonne, IL (United States)

    2014-12-10

    Advanced fast nuclear power plants and small modular fast reactors are composed of thin-walled structures such as pipes; as a result, they do not have sufficient inherent strength to resist seismic loads. Seismic isolation, therefore, is an effective solution for mitigating earthquake hazards for these types of structures. Base isolation, on which numerous studies have been conducted, is a well-defined structure protection system against earthquakes. In conventional isolators, such as high-damping rubber bearings, lead-rubber bearings, and friction pendulum bearings, large relative displacements occur between upper structures and foundations. Only isolation in a horizontal direction is provided; these features are not desirable for the piping systems. The concept of periodic materials, based on the theory of solid-state physics, can be applied to earthquake engineering. The periodic material is a material that possesses distinct characteristics that prevent waves with certain frequencies from being transmitted through it; therefore, this material can be used in structural foundations to block unwanted seismic waves with certain frequencies. The frequency band of periodic material that can filter out waves is called the band gap, and the structural foundation made of periodic material is referred to as the periodic foundation. The design of a nuclear power plant, therefore, can be unified around the desirable feature of a periodic foundation, while the continuous maintenance of the structure is not needed. In this research project, three different types of periodic foundations were studied: one-dimensional, two-dimensional, and three-dimensional. The basic theories of periodic foundations are introduced first to find the band gaps; then the finite element methods are used, to perform parametric analysis, and obtain attenuation zones; finally, experimental programs are conducted, and the test data are analyzed to verify the theory. This procedure shows that the

  3. Methods for the Isolation of Genes Encoding Novel PHA Metabolism Enzymes from Complex Microbial Communities.

    Science.gov (United States)

    Cheng, Jiujun; Nordeste, Ricardo; Trainer, Maria A; Charles, Trevor C

    2017-01-01

    Development of different PHAs as alternatives to petrochemically derived plastics can be facilitated by mining metagenomic libraries for diverse PHA cycle genes that might be useful for synthesis of bio-plastics. The specific phenotypes associated with mutations of the PHA synthesis pathway genes in Sinorhizobium meliloti and Pseudomonas putida, allows the use of powerful selection and screening tools to identify complementing novel PHA synthesis genes. Identification of novel genes through their function rather than sequence facilitates the functional proteins that may otherwise have been excluded through sequence-only screening methodology. We present here methods that we have developed for the isolation of clones expressing novel PHA metabolism genes from metagenomic libraries.

  4. Methods for the isolation of genes encoding novel PHB cycle enzymes from complex microbial communities.

    Science.gov (United States)

    Nordeste, Ricardo F; Trainer, Maria A; Charles, Trevor C

    2010-01-01

    Development of different PHAs as alternatives to petrochemically derived plastics can be facilitated by mining metagenomic libraries for diverse PHA cycle genes that might be useful for synthesis of bioplastics. The specific phenotypes associated with mutations of the PHA synthesis pathway genes in Sinorhizobium meliloti allows for the use of powerful selection and screening tools to identify complementing novel PHA synthesis genes. Identification of novel genes through their function rather than sequence facilitates finding functional proteins that may otherwise have been excluded through sequence-only screening methodology. We present here methods that we have developed for the isolation of clones expressing novel PHA metabolism genes from metagenomic libraries.

  5. Development of a structural model for the nonlinear shear deformation behavior of a seismic isolator

    International Nuclear Information System (INIS)

    Lee, Jae Han; Koo, Gyeong Hoi; Yoo, Bong

    2002-02-01

    The seismic excitation test results of an isolated test structure for artificial time history excitation are summarized for structure models of the isolated structure and isolation bearing. To simulate the response characteristic of isolated structure, shear hysteresis curves of isolators are analyzed. A simple analysis model is developed representing the actual dynamic behaviors of the test model, and the seismic responses using the simple model of the isolated structure and structure models, which are developed such as linear and bilinear models for isolators, are performed and compared with those of the seismic tests. The developed bilinear model is well applicable only to large shear strain area of LLRB

  6. [New isolation methods and phylogenetic diversity of actinobacteria from hypersaline beach in Aksu].

    Science.gov (United States)

    Zhang, Yao; Xia, Zhanfeng; Cao, Xinbo; Li, Jun; Zhang, Lili

    2013-08-04

    We explored 4 new methods to improve the isolation of actinobacterial resources from high salt areas. Optimized media based on 4 new strategies were used for isolating actinobacteria from hypersaline beaches. Glycerin-arginine, trehalose-creatine, glycerol-asparticacid, mannitol-casein, casein-mannitol, mannitol-alanine, chitosan-asparagineand GAUZE' No. 1 were used as basic media. New isolation strategy includes 4 methods: ten-fold dilution culture, simulation of the original environment, actinobacterial culture guided by uncultured molecular technology detected, and reference of actinobacterial media for brackish marine environment. The 16S rRNA genes of the isolates were amplified with bacterial universal primers. The results of 16S rRNA gene sequences were compared with sequences obtained from GenBank databases. We constructed phylogenetic tree with the neighbor-joining method. No actinobacterial strains were isolated by 8 media of control group, while 403 strains were isolated by new strategies. The isolates by new methods were members of 14 genera (Streptomyces, Streptomonospora, Saccharomonospora, Plantactinospora, Nocardia, Amycolatopsis, Glycomyces, Micromonospora, Nocardiopsis, Isoptericola, Nonomuraea, Thermobifida, Actinopolyspora, Actinomadura) of 10 families in 8 suborders. The most abundant and diverse isolates were the two suborders of Streptomycineae (69.96%) and Streptosporangineaesuborder (9.68%) within the phylum Actinobacteria, including 9 potential novel species. New isolation methods significantly improved the actinobacterial culturability of hypersaline areas, and obtained many potential novel species, which provided a new and more effective way to isolate actinobacteria resources in hypersaline environments.

  7. Development of inactivated-local isolate vaccine for infectious bronchitis

    Directory of Open Access Journals (Sweden)

    Darminto

    1999-06-01

    Full Text Available Infectious bronchitis (IB is an acute highly contagious viral respiratory disease of poultry caused by coronavirus. The disease causes high mortality in young chicks, reduce body weight gain in broilers and remarkable drop in egg production. IB can only be controlled by vaccination, but due to the antigenic variation among serotypes of IB viruses, the effective IB vaccine should be prepared from local isolates. The aim of this research is to develop inactivated IB vaccine derived from local IB isolates. Local isolates of IB viruses designated as I-37, I-269 and PTS-III were propagated respectively in specific pathogen free (SPF chicken eggs, the viruses then were inactivated by formaline at final concentration of 1:1,000. Subsequently, the inactivated viruses were mixed and emulsified in oil emulsion adjuvant with sorbitant mono-oleic as an emulsifier. The vaccine then was tested for its safety, potency and efficacy in broiler chickens. Birds inoculated twice with a two-week interval by inactivated vaccine did not show any adverse reaction, either systemic or local reaction. The inoculated birds developed antibody responses with high titre, while antibody of the control birds remain negative. In addition, efficacy test which was conducted in broilers demonstrated that birds vaccinated by live-commercial vaccine and boosted three weeks later by Balitvet inactivated vaccine showed high level of antibody production which provided high level of protection against challenged virus (76% against I-37, 92% against I-269 and 68% against PTS-III challenge viruses. From this study, it can be concluded that inactivated local IB vaccine is considered to be safe, potent and efficacious. The vaccine stimulates high titre of antibody responses, which provide high level of protection against challenged viruses.

  8. Methods for extracellular vesicles isolation in a hospital setting

    Directory of Open Access Journals (Sweden)

    Matías eSáenz-Cuesta

    2015-02-01

    Full Text Available The research in extracellular vesicles (EVs has been rising during the last decade. However, there is no clear consensus on the most accurate protocol to isolate and analyze them. Besides, most of the current protocols are difficult to implement in a hospital setting due to being very time consuming or to requirements of specific infrastructure. Thus, our aim is to compare five different protocols (comprising two different medium-speed differential centrifugation protocols; commercially polymeric precipitation -exoquick-; acid precipitation; and ultracentrifugation for blood and urine samples to determine the most suitable one for the isolation of EVs. Nanoparticle tracking analysis, flow cytometry, western blot, electronic microscopy and spectrophotometry were used to characterize basic aspects of EVs such us concentration, size distribution, cell-origin and transmembrane markers and RNA concentration. The highest EV concentrations were obtained using the exoquick protocol, followed by both differential centrifugation protocols, while the ultracentrifugation and acid-precipitation protocols yielded considerably lower EV concentrations. The five protocols isolated EVs of similar characteristics regarding markers and RNA concentration however standard protocol recovered only small EVs. EV isolated with exoquick presented difficult to be analyzed with western blot. The RNA concentrations obtained from urine-derived EVs were similar to those obtained from blood-derived ones, despite the urine EV concentration being 10 to 20 times lower. We consider that a medium-speed differential centrifugation could be suitable to be applied in a hospital setting due to require the simplest infrastructure and recover higher concentration of EV than standard protocol. A workflow from sampling to characterization of EVs is proposed.

  9. Intercomparison of analysis methods for seismically isolated nuclear structures. Part 1: Advanced test data and numerical methods. Working material

    International Nuclear Information System (INIS)

    1993-01-01

    The purpose of the meeting was to review proposed contributions from CRP participating organizations to discuss in detail the experimental data on seismic isolators, to review the numerical methods for the analysis of the seismic isolators, and to perform a first comparison of the calculation results. The aim of the CRP was to validate the reliable numerical methods used for both detailed evaluation of dynamic behaviour of isolation devices and isolated nuclear structures of different nuclear power plant types. The full maturity of seismic isolation for nuclear applications was stressed, as well as the excellent behaviour of isolated structures during the recent earthquakes in Japan and the USA. Participants from Italy, USA, Japan, Russian federation, Republic of Korea, United Kingdom, India and European Commission have presented overview papers on the present programs and their status of contribution to the CRP

  10. Intercomparison of analysis methods for seismically isolated nuclear structures. Part 1: Advanced test data and numerical methods. Working material

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1993-07-01

    The purpose of the meeting was to review proposed contributions from CRP participating organizations to discuss in detail the experimental data on seismic isolators, to review the numerical methods for the analysis of the seismic isolators, and to perform a first comparison of the calculation results. The aim of the CRP was to validate the reliable numerical methods used for both detailed evaluation of dynamic behaviour of isolation devices and isolated nuclear structures of different nuclear power plant types. The full maturity of seismic isolation for nuclear applications was stressed, as well as the excellent behaviour of isolated structures during the recent earthquakes in Japan and the USA. Participants from Italy, USA, Japan, Russian federation, Republic of Korea, United Kingdom, India and European Commission have presented overview papers on the present programs and their status of contribution to the CRP.

  11. Effect of starch isolation method on properties of sweet potato starch

    Directory of Open Access Journals (Sweden)

    A. SURENDRA BABU

    2014-08-01

    Full Text Available Isolation method of starch with different agents influences starch properties, which provide attention for studying the most appropriate method for isolation of starch. In the present study sweet potato starch was isolated by Sodium metabisulphate (M1, Sodium chloride (M2, and Distilled water (M3 methods and these were assessed for functional, chemical, pasting and structural properties. M3 yielded the greatest recovery of starch (10.20%. Isolation methods significantly changed swelling power and pasting properties but starches exhibited similar chemical properties. Sweet potato starches possessed C-type diffraction pattern. Small size granules of 2.90 μm were noticed in SEM of M3 starch. A high degree positive correlation was found between ash, amylose, and total starch content. The study concluded that isolation methods brought changes in yield, pasting and structural properties of sweet potato starch.

  12. How the RNA isolation method can affect microRNA microarray results

    DEFF Research Database (Denmark)

    Podolska, Agnieszka; Kaczkowski, Bogumil; Litman, Thomas

    2011-01-01

    RNA microarray analysis on porcine brain tissue. One method is a phenol-guanidine isothiocyanate-based procedure that permits isolation of total RNA. The second method, miRVana™ microRNA isolation, is column based and recovers the small RNA fraction alone. We found that microarray analyses give different results...... that depend on the RNA fraction used, in particular because some microRNAs appear very sensitive to the RNA isolation method. We conclude that precautions need to be taken when comparing microarray studies based on RNA isolated with different methods.......The quality of RNA is crucial in gene expression experiments. RNA degradation interferes in the measurement of gene expression, and in this context, microRNA quantification can lead to an incorrect estimation. In the present study, two different RNA isolation methods were used to perform micro...

  13. Simple and efficient method for isolation and cultivation of endoscopically obtained human colonocytes

    DEFF Research Database (Denmark)

    Seidelin, Jakob B; Horn, Thomas; Nielsen, Ole H

    2003-01-01

    and where the diagnosis of irritable bowel syndrome was later reached, were included. Seven colon biopsies were taken and incubated at varying time periods of 10-120 min and temperatures of 4-37 degrees C in a chelating buffer. The epithelium was then harvested and cultivated under three different......Few comparative and validated reports exist on the isolation and growth of colonoscopically obtained colonic epithelium. The aim of this study was to develop and validate a simple method for the cultivation of colonoscopically obtained colonocytes. Forty patients, who underwent routine colonoscopy...

  14. Failed sperm development as a reproductive isolating barrier between species.

    Science.gov (United States)

    Wünsch, Lisa K; Pfennig, Karin S

    2013-01-01

    Hybrid male sterility is a common reproductive isolating barrier between species. Yet, little is known about the actual developmental causes of this phenomenon, especially in naturally hybridizing species. We sought to evaluate the developmental causes of hybrid male sterility, using spadefoot toads as our study system. Plains spadefoot toads (Spea bombifrons) and Mexican spadefoot toads (S. multiplicata) hybridize where they co-occur in the southwestern USA. Hybrids are viable, but hybrid males suffer reduced fertility. We compared testes size and developmental stages of sperm cell maturation between hybrid males and males of each species. We found that testes of hybrid males did not differ in mean size from pure-species males. However, hybrids showed a greater range of within-individual variation in testes size than pure-species males. Moreover, although hybrids produced similar numbers of early stage sperm cells, hybrids produced significantly fewer mature spermatozoids than pure-species males. Interestingly, an introgressed individual produced numbers of live sperm comparable to pure-species males, but the majority of these sperm cells were abnormally shaped and non-motile. These results indicate that hybrid incompatibilities in late sperm development serve as a reproductive isolating barrier between species. The nature of this breakdown highlights the possibilities that hybrid males may vary in fertility and that fertility could possibly be recovered in introgressed males. © 2013 Wiley Periodicals, Inc.

  15. Methods for isolation and viability assessment of biological organisms

    Science.gov (United States)

    Letant, Sonia Edith; Baker, Sarah Elyse; Bond, Tiziana; Chang, Allan Shih-Ping

    2015-02-03

    Isolation of biological or chemical organisms can be accomplished using a surface enhanced Raman scattering (SERS) system. The SERS system can be a single or a stacked plurality of photonic crystal membranes with noble-metal lined through pores for flowing analyte potentially containing the biological or chemical organisms. The through pores can be adapted to trap individual biological or chemical organisms and emit SERS spectra, which can then be detected by a detector and further analyzed for viability of the biological or chemical organism.

  16. A rapid, efficient, and economic device and method for the isolation and purification of mouse islet cells.

    Directory of Open Access Journals (Sweden)

    Yin Zongyi

    Full Text Available Rapid, efficient, and economic method for the isolation and purification of islets has been pursued by numerous islet-related researchers. In this study, we compared the advantages and disadvantages of our developed patented method with those of commonly used conventional methods (Ficoll-400, 1077, and handpicking methods. Cell viability was assayed using Trypan blue, cell purity and yield were assayed using diphenylthiocarbazone, and islet function was assayed using acridine orange/ethidium bromide staining and enzyme-linked immunosorbent assay-glucose stimulation testing 4 days after cultivation. The results showed that our islet isolation and purification method required 12 ± 3 min, which was significantly shorter than the time required in Ficoll-400, 1077, and HPU groups (34 ± 3, 41 ± 4, and 30 ± 4 min, respectively; P 1000 islets. In summary, the MCT method is a rapid, efficient, and economic method for isolating and purifying murine islet cell clumps. This method overcomes some of the shortcomings of conventional methods, showing a relatively higher quality and yield of islets within a shorter duration at a lower cost. Therefore, the current method provides researchers with an alternative option for islet isolation and should be widely generalized.

  17. A method for isolating identifying and culturing of rat trachea-bronchia epithelial cells

    International Nuclear Information System (INIS)

    Cui Fengmei; Su Shibiao; Nie Jihua; Li Bingyan; Tong Jian

    2005-01-01

    Objective: To explore a method for isolating identifying and culturing the rat trachea-bronchia epithelial cells. Methods: The rat trachea-bronchia epithelial cells were isolated by digestion with pronase and brushing with cell brush, identified using confocul and cultured in entire F12 media with no serum. Results: With this method, cells in high purity and high viability could be obtained, and about 10 6 cells per rat. The cells grow well in entire F12 media with no serum. Conclusion: The method is useful for isolating rate trachea-bronchia epithelial cells and the entire F12 media with no serum is effective for culturing. (authors)

  18. Efficient methods for isolating five phytochemicals from Gentiana macrophylla using high-performance countercurrent chromatography.

    Science.gov (United States)

    Rho, Taewoong; Jung, Mila; Lee, Min Won; Chin, Young-Won; Yoon, Kee Dong

    2016-12-01

    Efficient high-performance countercurrent chromatography methods were developed to isolate five typical compounds from the extracts of Gentiana macrophylla. n-Butanol-soluble extract of G. macrophylla contained three hydrophilic iridoids, loganic acid (1), swertiamarin (2) and gentiopicroside (3), and a chromene derivative, macrophylloside D (4) which were successfully isolated by flow rate gradient (1.5 mL/min in 0-60 min, 5.0 mL/min in 60-120 min), and consecutive flow rate gradient HPCCC using n-butanol/0.1% aqueous trifluoroacetic acid (1:1, v/v, normal phase mode) system. The yields of 1-4 were 22, 16, 122, and 6 mg, respectively, with purities over 97% in a flow rate gradient high-performance countercurrent chromatography, and consecutive flow rate gradient high-performance countercurrent chromatography gave 1, 2, 3 (54, 41, 348 mg, respectively, purities over 97%) and 4 (13 mg, purity at 95%) from 750 mg of sample. The main compound in methylene chloride soluble extract, 2-methoxyanofinic acid, was successfully separated by n-hexane/ethyl acetate/methanol/water (4:6:4:6, v/v/v/v, flow-rate: 4 mL/min, reversed phase mode) condition. The structures of five isolates were elucidated by 1 H, 13 C NMR and ESI-Q-TOF-MS spectroscopic data which were compared with previously reported values. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  19. A novel method for isolation of epithelial cells from ovine esophagus for tissue engineering.

    Science.gov (United States)

    Macheiner, Tanja; Kuess, Anna; Dye, Julian; Saxena, Amulya K

    2014-01-01

    The yield of a critical number of basal epithelial cells with high mitotic rates from native tissue is a challenge in the field of tissue engineering. There are many protocols that use enzymatic methods for isolation of epithelial cells with unsatisfactory results for tissue engineering. This study aimed to develop a protocol for isolating a sufficient number of epithelial cells with a high Proliferating Index from ovine esophagus for tissue engineering applications. Esophageal mucosa was pretreated with dispase-collagenase solution and plated on collagen-coated culture dishes. Distinction of the various types of epithelial cells and developmental stages was done with specific primary antibodies to Cytokeratins and to Proliferating Cell Nuclear Antigen (PCNA). Up to approximately 8100 epithelial cells/mm2 of mucosa tissue were found after one week of migration. Cytokeratin 14 (CK 14) was positive identified in cells even after 83 days. At the same time the Proliferating Index was 71%. Our protocol for isolation of basal epithelial cells was successful to yield sufficient numbers of cells predominantly with proliferative character and without noteworthy negative enzymatic affection. The results at this study offer the possibility of generation critical cell numbers for tissue engineering applications.

  20. Stress and Deformation Analysis in Base Isolation Elements Using the Finite Element Method

    Directory of Open Access Journals (Sweden)

    Claudiu Iavornic

    2011-01-01

    Full Text Available In Modern tools as Finite Element Method can be used to study the behavior of elastomeric isolation systems. The simulation results obtained in this way provide a large series of data about the behavior of elastomeric isolation bearings under different types of loads and help in taking right decisions regarding geometrical optimizations needed for improve such kind of devices.

  1. Analysis of the error of the developed method of determination the active conductivity reducing the insulation level between one phase of the network and ground, and insulation parameters in a non-symmetric network with isolated neutral with voltage above 1000 V

    Science.gov (United States)

    Utegulov, B. B.

    2018-02-01

    In the work the study of the developed method was carried out for reliability by analyzing the error in indirect determination of the insulation parameters in an asymmetric network with an isolated neutral voltage above 1000 V. The conducted studies of the random relative mean square errors show that the accuracy of indirect measurements in the developed method can be effectively regulated not only by selecting a capacitive additional conductivity, which are connected between phases of the electrical network and the ground, but also by the selection of measuring instruments according to the accuracy class. When choosing meters with accuracy class of 0.5 with the correct selection of capacitive additional conductivity that are connected between the phases of the electrical network and the ground, the errors in measuring the insulation parameters will not exceed 10%.

  2. New method for the rapid extraction of natural products: efficient isolation of shikimic acid from star anise.

    Science.gov (United States)

    Just, Jeremy; Deans, Bianca J; Olivier, Wesley J; Paull, Brett; Bissember, Alex C; Smith, Jason A

    2015-05-15

    A new, practical, rapid, and high-yielding process for the pressurized hot water extraction (PHWE) of multigram quantities of shikimic acid from star anise (Illicium verum) using an unmodified household espresso machine has been developed. This operationally simple and inexpensive method enables the efficient and straightforward isolation of shikimic acid and the facile preparation of a range of its synthetic derivatives.

  3. Disturbances of dental development distinguish patients with oligodontia-ectodermal dysplasia from isolated oligodontia

    NARCIS (Netherlands)

    B. Dhamo (Brunilda); M.A.R. Kuijpers (Mette); Balk-Leurs, I. (I.); Boxum, C. (C.); E.B. Wolvius (Eppo); E.M. Ongkosuwito (Edwin)

    2017-01-01

    textabstractStructured Abstract: Objective: To investigate phenotypic differences in dental development between isolated oligodontia and oligodontia-ectodermal dysplasia (ED). Setting and sample population: A total of 129 patients diagnosed with isolated oligodontia and 22 patients with oligodontia

  4. A method to determine insulin responsiveness in synaptosomes isolated from frozen brain tissue.

    Science.gov (United States)

    Franklin, Whitney; Taglialatela, Giulio

    2016-03-01

    Studying the insulin signaling response at the synapse is an important approach to understand molecular mechanisms involved in disease-related neurodegenerative processes. We developed a method for studying the insulin responsiveness at the synaptic level by isolating functional synaptosomes from fresh or frozen tissue and exposing them to insulin in the presence of ATP (a critical step) to detect insulin receptor (IR) activation. We performed an ATP dose-response curve, insulin dose-response curve, and insulin response time course to optimize this method. We also demonstrated that our protocol reflects the degree of insulin responsiveness in vivo by using an animal model of known insulin resistance, AtENPP1-Tg mice. This method is advantageous over other methods detecting IR in total brain homogenates due to the ability to detect IR response without confounding contributions from other cell areas and cell types also expressing IR. Furthermore, ex vivo insulin stimulation can be compared to baseline synaptosomes obtained from the same animal which improves reliability and statistical power while decreasing the number of animals required to perform individual experiments. We have developed a reliable, efficient method to measure insulin-driven ex vivo phosphorylation of the synaptosomal insulin receptor that can reliably reflect the pre-existing insulin responsiveness status in the CNS of the animal. To the best of our knowledge, this is the first evidence of stimulation of isolated synaptosomes with insulin and a promising new technique to study the synaptic CNS insulin responsiveness under physiological or disease conditions. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. In vitro culture of functionally active buffalo hepatocytes isolated by using a simplified manual perfusion method.

    Directory of Open Access Journals (Sweden)

    Santanu Panda

    Full Text Available In farm animals, there is no suitable cell line available to understand liver-specific functions. This has limited our understanding of liver function and metabolism in farm animals. Culturing and maintenance of functionally active hepatocytes is difficult, since they survive no more than few days. Establishing primary culture of hepatocytes can help in studying cellular metabolism, drug toxicity, hepatocyte specific gene function and regulation. Here we provide a simple in vitro method for isolation and short-term culture of functionally active buffalo hepatocytes.Buffalo hepatocytes were isolated from caudate lobes by using manual enzymatic perfusion and mechanical disruption of liver tissue. Hepatocyte yield was (5.3 ± 0.66×107 cells per gram of liver tissue with a viability of 82.3 ± 3.5%. Freshly isolated hepatocytes were spherical with well contrasted border. After 24 hours of seeding onto fibroblast feeder layer and different extracellular matrices like dry collagen, matrigel and sandwich collagen coated plates, hepatocytes formed confluent monolayer with frequent clusters. Cultured hepatocytes exhibited typical cuboidal and polygonal shape with restored cellular polarity. Cells expressed hepatocyte-specific marker genes or proteins like albumin, hepatocyte nuclear factor 4α, glucose-6-phosphatase, tyrosine aminotransferase, cytochromes, cytokeratin and α1-antitrypsin. Hepatocytes could be immunostained with anti-cytokeratins, anti-albumin and anti α1-antitrypsin antibodies. Abundant lipid droplets were detected in the cytosol of hepatocytes using oil red stain. In vitro cultured hepatocytes could be grown for five days and maintained for up to nine days on buffalo skin fibroblast feeder layer. Cultured hepatocytes were viable for functional studies.We developed a convenient and cost effective technique for hepatocytes isolation for short-term culture that exhibited morphological and functional characteristics of active hepatocytes

  6. Improved Simplified Methods for Effective Seismic Analysis and Design of Isolated and Damped Bridges in Western and Eastern North America

    Science.gov (United States)

    Koval, Viacheslav

    The seismic design provisions of the CSA-S6 Canadian Highway Bridge Design Code and the AASHTO LRFD Seismic Bridge Design Specifications have been developed primarily based on historical earthquake events that have occurred along the west coast of North America. For the design of seismic isolation systems, these codes include simplified analysis and design methods. The appropriateness and range of application of these methods are investigated through extensive parametric nonlinear time history analyses in this thesis. It was found that there is a need to adjust existing design guidelines to better capture the expected nonlinear response of isolated bridges. For isolated bridges located in eastern North America, new damping coefficients are proposed. The applicability limits of the code-based simplified methods have been redefined to ensure that the modified method will lead to conservative results and that a wider range of seismically isolated bridges can be covered by this method. The possibility of further improving current simplified code methods was also examined. By transforming the quantity of allocated energy into a displacement contribution, an idealized analytical solution is proposed as a new simplified design method. This method realistically reflects the effects of ground-motion and system design parameters, including the effects of a drifted oscillation center. The proposed method is therefore more appropriate than current existing simplified methods and can be applicable to isolation systems exhibiting a wider range of properties. A multi-level-hazard performance matrix has been adopted by different seismic provisions worldwide and will be incorporated into the new edition of the Canadian CSA-S6-14 Bridge Design code. However, the combined effect and optimal use of isolation and supplemental damping devices in bridges have not been fully exploited yet to achieve enhanced performance under different levels of seismic hazard. A novel Dual-Level Seismic

  7. Development of guidelines for seismic isolation in Italy

    International Nuclear Information System (INIS)

    Olivieri, M.; Martelli, A.; Bettinali, F.; Bonacina, G.

    1992-01-01

    The first activities on seismic isolation that were performed in Italy concerned the preparation of a proposal for design guidelines for nuclear power plants using the high damping steel-laminated elastomer bearings (HDLRBs). They were jointly initiated by ENEA-RIN and GE Nuclear Energy in 1988, with the co-operation of ISMES and the support of experts of ENEA-DISP and Bechtel National Inc. The features of the guidelines proposal were outlined at the First Post-SMiRT Conference Seminar on Seismic Base Isolation of Nuclear Power Facilities (San Francisco, 1989). The full text of the document was published in the Journal 'Energia Nucleare' in 1990, in a tentative form, to allow for a broad review. A summary of the main items - together with some first results of R and D studies performed in support to guidelines development - was also reported in a paper which was recently published by the Journal 'Nuclear Technology' (February 1992). A first revision of the document is being prepared and will be soon published: it accounts for both comments received - for instance, by the American Society of Civil Engineers (ASCE), ENEA-DISP and the Malaysian Rubber Producers' Association (MRPRA) - and the first results of R and D studies in progress in Italy and the USA. These activities have recently been extended - as part of a cooperation with the Italian Standard Authority (UNI) - to other antiseismic devices, for application to civil buildings and non-nuclear plants. A co-operation of ENEA, ENEL and ISMES has also been started with the National Seismic Service to help it in the assessment of national regulations. Furthermore, extension of the aforesaid guidelines document to nuclear reactors using bearings different from the HDLRB has been planned, under the sponsorship of the Commission of the European communities: this work will be performed by ENEA, with the cooperation of ALGA, ISMES, ANSALDO and the Nuclear Engineering Laboratory (LIN) of the Bologna University, and the

  8. Novel tandem column method for the rapid isolation of radiostrontium from human urine

    International Nuclear Information System (INIS)

    Hawkins, Cory A.; Shkrob, Ilya A.; Mertz, Carol J.; Dietz, Mark L.; Kaminski, Michael D.

    2012-01-01

    Highlights: ► Method for separation and preconcentration of radiostrontium from human urine. ► Recoveries >98%, concentration factor of ca. 50, processing time of nearly 1 h. ► Retention model developed to assist optimization of separations on Diphonix ® column. ► Semi-automated sample preparation device developed. - Abstract: A method has been developed for the isolation of strontium from human urine for subsequent determination in sample volumes as low as 5–20 mL. This method involves the acidification of the sample using methanesulfonic acid and its decolorization using charcoal, treatment of the filtrate with Diphonix ® resin, and subsequent concentration of strontium on Sr resin. Data from retention model simulations provided the initial conditions which were then optimized by actual column separations. Diphonix ® resin was shown to be effective at removing alkali metal ions from the urine matrix under conditions that retain higher valence ions. The suggested processing method provides 99% recovery of Sr 2+ , a concentration factor of 50, and an expected per sample processing time of less than 1 h.

  9. An Improved Quick Method for the Isolation of Total RNA from Cotton ...

    African Journals Online (AJOL)

    David PANG

    2011-11-02

    Nov 2, 2011 ... in liquid nitrogen in a mortar and pestle and stored until. RNA isolation. ... our laboratory for microarray analysis, cDNA pyro- sequencing studies and construction ..... Economic and rapid method for extracting cotton genomic ...

  10. iPBS: a universal method for DNA fingerprinting and retrotransposon isolation.

    Science.gov (United States)

    Kalendar, Ruslan; Antonius, Kristiina; Smýkal, Petr; Schulman, Alan H

    2010-11-01

    Molecular markers are essential in plant and animal breeding and biodiversity applications, in human forensics, and for map-based cloning of genes. The long terminal repeat (LTR) retrotransposons are well suited as molecular markers. As dispersed and ubiquitous transposable elements, their "copy and paste" life cycle of replicative transposition leads to new genome insertions without excision of the original element. Both the overall structure of retrotransposons and the domains responsible for the various phases of their replication are highly conserved in all eukaryotes. Nevertheless, up to a year has been required to develop a retrotransposon marker system in a new species, involving cloning and sequencing steps as well as the development of custom primers. Here, we describe a novel PCR-based method useful both as a marker system in its own right and for the rapid isolation of retrotransposon termini and full-length elements, making it ideal for "orphan crops" and other species with underdeveloped marker systems. The method, iPBS amplification, is based on the virtually universal presence of a tRNA complement as a reverse transcriptase primer binding site (PBS) in LTR retrotransposons. The method differs from earlier retrotransposon isolation methods because it is applicable not only to endogenous retroviruses and retroviruses, but also to both Gypsy and Copia LTR retrotransposons, as well as to non-autonomous LARD and TRIM elements, throughout the plant kingdom and to animals. Furthermore, the inter-PBS amplification technique as such has proved to be a powerful DNA fingerprinting technology without the need for prior sequence knowledge.

  11. Cell fusion in tumor progression: the isolation of cell fusion products by physical methods

    Directory of Open Access Journals (Sweden)

    Vincitorio Massimo

    2011-09-01

    Full Text Available Abstract Background Cell fusion induced by polyethylene glycol (PEG is an efficient but poorly controlled procedure for obtaining somatic cell hybrids used in gene mapping, monoclonal antibody production, and tumour immunotherapy. Genetic selection techniques and fluorescent cell sorting are usually employed to isolate cell fusion products, but both procedures have several drawbacks. Results Here we describe a simple improvement in PEG-mediated cell fusion that was obtained by modifying the standard single-step procedure. We found that the use of two PEG undertreatments obtains a better yield of cell fusion products than the standard method, and most of these products are bi- or trinucleated polykaryocytes. Fusion rate was quantified using fluorescent cell staining microscopy. We used this improved cell fusion and cell isolation method to compare giant cells obtained in vitro and giant cells obtained in vivo from patients with Hodgkin's disease and erythroleukemia. Conclusions In the present study we show how to improve PEG-mediated cell fusion and that cell separation by velocity sedimentation offers a simple alternative for the efficient purification of cell fusion products and to investigate giant cell formation in tumor development.

  12. Base Isolation for Seismic Retrofitting of a Multiple Building Structure: Evaluation of Equivalent Linearization Method

    Directory of Open Access Journals (Sweden)

    Massimiliano Ferraioli

    2016-01-01

    Full Text Available Although the most commonly used isolation systems exhibit nonlinear inelastic behaviour, the equivalent linear elastic analysis is commonly used in the design and assessment of seismic-isolated structures. The paper investigates if the linear elastic model is suitable for the analysis of a seismically isolated multiple building structure. To this aim, its computed responses were compared with those calculated by nonlinear dynamic analysis. A common base isolation plane connects the isolation bearings supporting the adjacent structures. In this situation, the conventional equivalent linear elastic analysis may have some problems of accuracy because this method is calibrated on single base-isolated structures. Moreover, the torsional characteristics of the combined system are significantly different from those of separate isolated buildings. A number of numerical simulations and parametric studies under earthquake excitations were performed. The accuracy of the dynamic response obtained by the equivalent linear elastic model was calculated by the magnitude of the error with respect to the corresponding response considering the nonlinear behaviour of the isolation system. The maximum displacements at the isolation level, the maximum interstorey drifts, and the peak absolute acceleration were selected as the most important response measures. The influence of mass eccentricity, torsion, and high-modes effects was finally investigated.

  13. A historical review of Waste Isolation Pilot Plant backfill development

    International Nuclear Information System (INIS)

    Krumhansl, James L.; Molecke, Martin A.; Papenguth, Hans W.; Brush, Laurence H.

    2000-01-01

    Backfills have been part of Sandia National Laboratories' [Sandia's] Waste Isolation Pilot Plant [WIPP] designs for over twenty years. Historically, backfill research at Sandia has depended heavily on the changing mission of the WIPP facility. Early testing considered heat producing, high level, wastes. Bentonite/sand/salt mixtures were evaluated and studies focused on developing materials that would retard brine ingress, sorb radionuclides, and withstand elevated temperatures. The present-day backfill consists of pure MgO [magnesium oxide] in a pelletized form and is directed at treating the relatively low contamination level, non-heat producing, wastes actually being disposed of in the WIPP. Its introduction was motivated by the need to scavenging CO 2 [carbon dioxide] from decaying organic components in the waste. However, other benefits, such as a substantial desiccating capacity, are also being evaluated. The MgO backfill also fulfills a statutory requirement for assurance measures beyond those needed to demonstrate compliance with the US Environmental Protection Agency [EPA] regulatory release limits. However, even without a backfill, the WIPP repository design still operates within EPA regulatory release limits

  14. Formation of putative chloroplast cytochromes in isolated developing pea chloroplasts

    International Nuclear Information System (INIS)

    Thaver, S.S.; Bhava, D.; Castelfranco, P.A.

    1986-01-01

    In addition to chlorophyll-protein complexes, other proteins were labeled when isolated developing pea chloroplasts were incubated with [ 14 C]-5-aminolevulinic acid [ 14 C]-ALA. The major labeled band (M/sub r/ = 43 kDa by LDS-PAGE) was labeled even in the presence of chloramphenicol. Heme-dependent peroxidase activity (as detected by the tetramethyl benzidine-H 2 O 2 stain) was not visibly associated with this band. The radioactive band was stable to heat, 5% HCl in acetone, and was absent if the incubation with [ 14 C]-5-aminolevulinic acid was carried out in the presence of N-methyl protoporphyrin IX dimethyl ester (a specific inhibitor of ferrochelatase). Organic solvent extraction procedures for the enrichment of cytochrome f from chloroplast membranes also extracted this unknown labeled product. It was concluded that this labeled product was probably a c-type cytochrome. The effect of exogenous iron, iron chelators, gabaculine (an inhibitor of ALA synthesis) and other incubation conditions upon the in vitro formation of putative chloroplast cytochromes will be discussed

  15. Test of a mosquito eggshell isolation method and subsampling procedure.

    Science.gov (United States)

    Turner, P A; Streever, W J

    1997-03-01

    Production of Aedes vigilax, the common salt-marsh mosquito, can be assessed by determining eggshell densities found in soil. In this study, 14 field-collected eggshell samples were used to test a subsampling technique and compare eggshell counts obtained with a flotation method to those obtained by direct examination of sediment (DES). Relative precision of the subsampling technique was assessed by determining the minimum number of subsamples required to estimate the true mean and confidence interval of a sample at a predetermined confidence level. A regression line was fitted to cube-root transformed eggshell counts obtained from flotation and DES and found to be significant (P eggshells present. Eggshells obtained with the flotation method can be used to predict those from DES using the following equation: DES count = [1.386 x (flotation count)0.33 - 0.01]3.

  16. Extraction methods of Amaranthus sp. grain oil isolation.

    Science.gov (United States)

    Krulj, Jelena; Brlek, Tea; Pezo, Lato; Brkljača, Jovana; Popović, Sanja; Zeković, Zoran; Bodroža Solarov, Marija

    2016-08-01

    Amaranthus sp. is a fast-growing crop with well-known beneficial nutritional values (rich in protein, fat, dietary fiber, ash, and minerals, especially calcium and sodium, and containing a higher amount of lysine than conventional cereals). Amaranthus sp. is an underexploited plant source of squalene, a compound of high importance in the food, cosmetic and pharmaceutical industries. This paper has examined the effects of the different extraction methods (Soxhlet, supercritical fluid and accelerated solvent extraction) on the oil and squalene yield of three genotypes of Amaranthus sp. grain. The highest yield of the extracted oil (78.1 g kg(-1) ) and squalene (4.7 g kg(-1) ) in grain was obtained by accelerated solvent extraction (ASE) in genotype 16. Post hoc Tukey's HSD test at 95% confidence limit showed significant differences between observed samples. Principal component analysis (PCA) and cluster analysis (CA) were used for assessing the effect of different genotypes and extraction methods on oil and squalene yield, and also the fatty acid composition profile. Using coupled PCA and CA of observed samples, possible directions for improving the quality of product can be realized. The results of this study indicate that it is very important to choose both the right genotype and the right method of extraction for optimal oil and squalene yield. © 2015 Society of Chemical Industry. © 2015 Society of Chemical Industry.

  17. Development of a novel vaccine against canine parvovirus infection with a clinical isolate of the type 2b strain

    OpenAIRE

    Park, Seon Ah; Park, Seung-Yong; Song, Chang-Seon; Choi, In-Soo; Kim, Hwi Yool; Lee, Joong-Bok; Lee, Nak-Hyung

    2012-01-01

    Purpose In spite of an extensive vaccination program, parvoviral infections still pose a major threat to the health of dogs. Materials and Methods We isolated a novel canine parvovirus (CPV) strain from a dog with enteritis. Nucleotide and amino acid sequence analysis of the isolate showed that it is a novel type 2b CPV with asparagine at the 426th position and valine at the 555th position in VP2. To develop a vaccine against CPV infection, we passaged the isolate 4 times in A72 cells. Result...

  18. A novel affinity purification method to isolate peptide specific antibodies

    DEFF Research Database (Denmark)

    Karlsen, Alan E; Lernmark, A; Kofod, Hans

    1990-01-01

    Site-specific, high affinity polyclonal antisera are effectively and successfully produced by immunizing rabbits with synthetic peptides. The use of these antisera in subsequent immune analysis is often limited because of non-specific binding. We describe a new and simple method to effectively...... affinity-purify anti-peptide antibodies. To test our system, rabbits were immunized with model peptides representing sequences of the putative rabbit growth hormone receptor and several HLA-DQ beta-chain molecules. Polystyrene plastic beads were coated with peptides. Immune serum was incubated...... with the beads and after a wash step the bound antibodies were eluted in 1 M acetic acid. The eluted material was composed predominantly of intact immunoglobulin as evidenced by the presence of heavy and light chain bands in SDS-PAGE. The eluted antibodies were peptide specific in ELISA and bound only to intact...

  19. Automated methods for single-stranded DNA isolation and dideoxynucleotide DNA sequencing reactions on a robotic workstation

    International Nuclear Information System (INIS)

    Mardis, E.R.; Roe, B.A.

    1989-01-01

    Automated procedures have been developed for both the simultaneous isolation of 96 single-stranded M13 chimeric template DNAs in less than two hours, and for simultaneously pipetting 24 dideoxynucleotide sequencing reactions on a commercially available laboratory workstation. The DNA sequencing results obtained by either radiolabeled or fluorescent methods are consistent with the premise that automation of these portions of DNA sequencing projects will improve the reproducibility of the DNA isolation and the procedures for these normally labor-intensive steps provides an approach for rapid acquisition of large amounts of high quality, reproducible DNA sequence data

  20. Method Engineering: Engineering of Information Systems Development Methods and Tools

    NARCIS (Netherlands)

    Brinkkemper, J.N.; Brinkkemper, Sjaak

    1996-01-01

    This paper proposes the term method engineering for the research field of the construction of information systems development methods and tools. Some research issues in method engineering are identified. One major research topic in method engineering is discussed in depth: situational methods, i.e.

  1. METHODS TO DEVELOP A TOROIDAL SURFACE

    Directory of Open Access Journals (Sweden)

    DANAILA Ligia

    2017-05-01

    Full Text Available The paper work presents two practical methods to draw the development of a surface unable to be developed applying classical methods of Descriptive Geometry, the toroidal surface, frequently met in technical practice. The described methods are approximate ones; the development is obtained with the help of points. The accuracy of the methods is given by the number of points used when drawing. As for any other approximate method, when practically manufactured the development may need to be adjusted on site.

  2. Development of seismic isolation system in vertical direction

    International Nuclear Information System (INIS)

    Ohoka, Makoto; Horikiri, Morito

    1999-04-01

    A structure concept of vertical seismic isolation system which uses a common deck and a set of large dish springs was created in past studies. In this report, a series of dynamic tests on a small scale model of a common deck isolation structure were performed. The model was excited by random and seismic waves in the horizontal direction and 2-D excitation, horizontal and vertical, in order to identify the characteristics of isolation effect. The tests results are summarized as below. 1) This structure has three vibration mode. The second mode is rocking. 2) Rocking frequency depends on the excitation, for this structure has dish spring which contact with cylinders. Rocking damping varies from 2 to 8%, 3) Each mode's response peak frequency to 2-D(horizontal and vertical) excitation is almost the same the some to horizontal excitation. Vertical mode damping to 2-D excitation is about three times to horizontal excitation. 4) Isolation effect depends on a characteristics of frequency of input motion. The minimum response is to the Monju design seismic wave, soil shear wave:Vs=2000 m/sec, natural frequency of horizontal isolation in vertical direction:fv=20 Hz. A relative displacement is controlled. 5) A rocking angular displacement to 2-D excitation is about 2 times to 1-D excitation(vertical). However, it is about 1.2 E-4(rad), sufficiently small for a practical plant. (author)

  3. Methods for the evaluation of antibiotic resistance in Lactobacillus isolated from fermented sausages

    Directory of Open Access Journals (Sweden)

    Hanna Lethycia Wolupeck

    Full Text Available ABSTRACT: The present study aimed to assess the antibiotic resistance in 54 indigenous Lactobacillus plantarum isolated from artisanal fermented sausages. The confirmation of the strain species was performed by multiplex-PCR assay. Antibiotic resistance was assessed by disk diffusion (DD and Minimum Inhibitory Concentration (MIC methods. Of 54 L. plantarum, 44 strains were genotypically confirmed as L. plantarum and 3 as Lactobacillus pentosus. The highest resistance rates were to ampicillin and streptomycin. The highest susceptibility rates were shown to tetracycline, chloramphenicol and penicillin G. None of the strains showed multidrug resistance. Resistance rates by DD and MIC were not different (P>0.05 for ampicillin, chloramphenicol, erythromycin and penicillin G. Future research should assess the genetic mechanisms underlying the phenotypic resistance in Lactobacillus strains to screen the potential probiotic strains for the development of functional meat products.

  4. [Improved methods for researching isolated carotid sinus baroreceptors automatically controlling for sinus pressure].

    Science.gov (United States)

    Wei, Hua; Zhao, Hai-Yan; Liu, Ping; Huang, Hai-Xia; Wang, Wei; Fu, Xiao-Suo; Niu, Wei-Zhen

    2013-01-01

    To develop a system for automatically controlling carotid sinus pressure in the study on baroreceptors. The preparation containing carotid sinus with parts of the connected vessels and carotid sinus nerve (CS-CSN) were isolated and perfused. A critical pressure controlling component (PRE-U, Hoerbiger, Deutschland) dictated by a computer was integrated into the system to clamp the intrasinus pressure. The pressure command and the relevant intrasinus pressure were compared to evaluate the validity of the pressure controlling system. A variety of sinus pressure-controlling patterns, including pulsation, ramp and step pressures, could be achieved accurately by using the system, and the pressure-dependent discharge activities of sinus nerve were confirmed. This system for clamping carotid sinus pressure could realize multiple pressure-controlling patterns and is a useful and flexible pressure controlling method that could applied in the study on mechano-electric transduction of baroreceptors.

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

    Science.gov (United States)

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

    1996-04-12

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

  6. Improved method for isolation of lycopsamine from roots of comfrey (Symphytum officinale).

    Science.gov (United States)

    Janes, Damjan; Kalamar, Bostjan; Kreft, Samo

    2012-07-01

    An improved method for the isolation and purification of pyrrolizidine alkaloids from comfrey (Symphytum officinale L.) roots was developed, introducing very fast, selective and ion residue-free reduction of N-oxides followed by ion-exchange chromatography giving a non-aqueous solution of alkaloids, from which solvents can be easily removed. With this procedure the use of large volumes of organic solvents, very slow reduction of N-oxides and input of additional impurities was avoided. Lycopsamine, which proved to be the major alkaloid, was additionally purified by preparative layer chromatography (PLC) and high performance liquid chromatography (HPLC). The identity of the alkaloid was confirmed by (I)H NMR spectroscopy and mass spectrometry.

  7. A rapid and inexpensive method for isolation of total DNA from Trichoderma spp (Hypocreaceae).

    Science.gov (United States)

    Vazquez-Angulo, J C; Mendez-Trujillo, V; González-Mendoza, D; Morales-Trejo, A; Grimaldo-Juarez, O; Cervantes-Díaz, L

    2012-05-15

    Extraction of high-quality genomic DNA for PCR amplification from filamentous fungi is difficult because of the complex cell wall and the high concentrations of polysaccharides and other secondary metabolites that bind to or co-precipitate with nucleic acids. We developed a modified sodium dodecyl sulfate/phenol protocol, without maceration in liquid nitrogen and without a final ethanol precipitation step. The A(260/280) absorbance ratios of isolated DNA were approximately 1.7-1.9, demonstrating that the DNA fraction is pure and can be used for analysis. Additionally, the A(260/230) values were higher than 1.6, demonstrating negligible contamination by polysaccharides. The DNA isolated by this protocol is of sufficient quality for molecular applications; this technique could be applied to other organisms that have similar substances that hinder DNA extraction. The main advantages of the method are that the mycelium is directly recovered from culture medium and it does not require the use of expensive and specialized equipment.

  8. Technical note: A method for isolating glycogen granules from ruminal protozoa for further characterization.

    Science.gov (United States)

    Hall, Mary Beth

    2016-03-01

    Evaluation of physical, chemical, and enzymatic hydrolysis characteristics of protozoal glycogen is best performed on a pure substrate to avoid interference from other cell components. A method for isolating protozoal glycogen granules without use of detergents or other potentially contaminating chemicals was developed. Rumen inoculum was incubated anerobically in vitro with glucose. Glycogen-laden protozoa produced in the fermentation, primarily isotrichids, were allowed to sediment in a separatory funnel and were dispensed. The protozoa were processed through repeated centrifugations and sonication to isolate glycogen granules largely free of feed and cellular debris. The final water-insoluble lyophilized product analyzed as 98.3% α-glucan with very rare starch granules and 1.9% protein. Observed losses of glycogen granules during the clean-up process indicate that this procedure should not be used for quantitative assessment of protozoal glycogen from fermentations. Further optimization of this procedure to enhance the amount of glycogen obtained per fermentation may be possible. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  9. Development and characterization of a magnetorheological elastomer based adaptive seismic isolator

    International Nuclear Information System (INIS)

    Li, Yancheng; Li, Jianchun; Samali, Bijan; Li, Weihua

    2013-01-01

    One of the main shortcomings in current base isolation design/practice is lack of adaptability. As a result, a base isolation system that is effective for one type earthquake may become ineffective or may have adverse effect for other earthquakes. The vulnerability of traditional base isolation systems can be exaggerated by two types of earthquakes, i.e. near-field earthquakes and far-field earthquakes. This paper addresses the challenge facing current base isolation design/practice by proposing a new type of seismic isolator for the base isolation system, namely an adaptive seismic isolator. The novel adaptive seismic isolator utilizes magnetorheological elastomer (MRE) for its field-sensitive material property. Traditional seismic isolator design with a unique laminated structure of steel and MRE layers has been adopted in the novel MRE seismic isolator. To evaluate and characterize the behavior of the MRE seismic isolator, experimental testing was conducted on a shake table facility under harmonic cycling loading. Experimental results show that the proposed adaptive seismic isolator can successfully alter the lateral stiffness and damping force in real time up to 37% and 45% respectively. Based on the successful development of the novel adaptive seismic isolator, a discussion is also extended to the impact and potential applications of such a device in structural control applications in civil engineering. (paper)

  10. Characterization of RNA from Exosomes and Other Extracellular Vesicles Isolated by a Novel Spin Column-Based Method

    Science.gov (United States)

    Enderle, Daniel; Spiel, Alexandra; Coticchia, Christine M.; Berghoff, Emily; Mueller, Romy; Schlumpberger, Martin; Sprenger-Haussels, Markus; Shaffer, Jonathan M.; Lader, Eric; Skog, Johan; Noerholm, Mikkel

    2015-01-01

    Exosomes and other extracellular vesicles (commonly referred to as EVs) have generated a lot of attention for their potential applications in both diagnostics and therapeutics. The contents of these vesicles are the subject of intense research, and the relatively recent discovery of RNA inside EVs has raised interest in the biological function of these RNAs as well as their potential as biomarkers for cancer and other diseases. Traditional ultracentrifugation-based protocols to isolate EVs are labor-intensive and subject to significant variability. Various attempts to develop methods with robust, reproducible performance have not yet been completely successful. Here, we report the development and characterization of a spin column-based method for the isolation of total RNA from EVs in serum and plasma. This method isolates highly pure RNA of equal or higher quantity compared to ultracentrifugation, with high specificity for vesicular over non-vesicular RNA. The spin columns have a capacity to handle up to 4 mL sample volume, enabling detection of low-abundance transcripts in serum and plasma. We conclude that the method is an improvement over traditional methods in providing a faster, more standardized way to achieve reliable high quality RNA preparations from EVs in biofluids such as serum and plasma. The first kit utilizing this new method has recently been made available by Qiagen as “exoRNeasy Serum/Plasma Maxi Kit”. PMID:26317354

  11. Evaluating Methods for Isolating Total RNA and Predicting the Success of Sequencing Phylogenetically Diverse Plant Transcriptomes

    Science.gov (United States)

    Bruskiewich, Richard; Burris, Jason N.; Carrigan, Charlotte T.; Chase, Mark W.; Clarke, Neil D.; Covshoff, Sarah; dePamphilis, Claude W.; Edger, Patrick P.; Goh, Falicia; Graham, Sean; Greiner, Stephan; Hibberd, Julian M.; Jordon-Thaden, Ingrid; Kutchan, Toni M.; Leebens-Mack, James; Melkonian, Michael; Miles, Nicholas; Myburg, Henrietta; Patterson, Jordan; Pires, J. Chris; Ralph, Paula; Rolf, Megan; Sage, Rowan F.; Soltis, Douglas; Soltis, Pamela; Stevenson, Dennis; Stewart, C. Neal; Surek, Barbara; Thomsen, Christina J. M.; Villarreal, Juan Carlos; Wu, Xiaolei; Zhang, Yong; Deyholos, Michael K.; Wong, Gane Ka-Shu

    2012-01-01

    Next-generation sequencing plays a central role in the characterization and quantification of transcriptomes. Although numerous metrics are purported to quantify the quality of RNA, there have been no large-scale empirical evaluations of the major determinants of sequencing success. We used a combination of existing and newly developed methods to isolate total RNA from 1115 samples from 695 plant species in 324 families, which represents >900 million years of phylogenetic diversity from green algae through flowering plants, including many plants of economic importance. We then sequenced 629 of these samples on Illumina GAIIx and HiSeq platforms and performed a large comparative analysis to identify predictors of RNA quality and the diversity of putative genes (scaffolds) expressed within samples. Tissue types (e.g., leaf vs. flower) varied in RNA quality, sequencing depth and the number of scaffolds. Tissue age also influenced RNA quality but not the number of scaffolds ≥1000 bp. Overall, 36% of the variation in the number of scaffolds was explained by metrics of RNA integrity (RIN score), RNA purity (OD 260/230), sequencing platform (GAIIx vs HiSeq) and the amount of total RNA used for sequencing. However, our results show that the most commonly used measures of RNA quality (e.g., RIN) are weak predictors of the number of scaffolds because Illumina sequencing is robust to variation in RNA quality. These results provide novel insight into the methods that are most important in isolating high quality RNA for sequencing and assembling plant transcriptomes. The methods and recommendations provided here could increase the efficiency and decrease the cost of RNA sequencing for individual labs and genome centers. PMID:23185583

  12. Scenario development methods and practice

    International Nuclear Information System (INIS)

    2001-01-01

    The safe management of radioactive waste is an essential aspect of all nuclear power programmes. Although a general consensus has been reached in OECD countries on the use of geological repositories for the disposal of high-level radioactive waste, analysis of the long-term safety of these repositories, using performance assessment and other tools, is required prior to implementation. The initial stage in developing a repository safety assessment is the identification of all factors that may be relevant to the long-term safety of the repository and their combination to form scenarios. This must be done in a systematic and transparent way in order to assure the regulatory authorities that nothing important has been forgotten. Scenario development has become the general term used to describe the collection and organisation of the scientific and technical information necessary to assess the long-term performance or safety of radioactive waste disposal systems. This includes the identification of the relevant features, events and processes (FEPs), the synthesis of broad models of scientific understanding, and the selection of cases to be calculated. Scenario development provides the overall framework in which the cases and their calculated consequences can be discussed, including biases or shortcomings due to omissions or lack of knowledge. The NEA Workshop on Scenario Development was organised in Madrid, in May 1999, with the objective of reviewing developments in scenario methodologies and applications in safety assessments since 1992. The outcome of this workshop is the subject of this book. It is a review of developments in scenario methodologies based on a large body of practical experience in safety assessments. It will be of interest to radioactive waste management experts as well as to other specialists involved in the development of scenario methodologies. (author)

  13. Development of Improved Process with Treatment of Cellulase for Isolation of Ampelopsin from Dried Fruits of Ampelopsis grossedentata

    Directory of Open Access Journals (Sweden)

    Wa Gao

    2016-02-01

    Full Text Available The commercial method for isolation of ampelopsin, one of the most common flavonoids isolated from the plant species Ampelopsis grossedentata, is a simple hydrothermal extraction at high temperature. To develop an improved process to isolate ampelopsin, the effects of treatment of cellulase on hydrolysis of the dried fruit of A. grossedentata were investigated. The treatment of cellulase was found to decrease the temperature and time for hydrolysis of the dried fruit of A. grossedentata. The conditions of the filter press and continuous flow centrifuge for removal of insoluble materials from the hydrolysate of the dried fruit of A. grossedentata were optimized. The recovery yield of ampelopsin from the dried fruits of A. grossedentata was 39.4%, as determined by HPLC chromatographic analysis. A safe and economical process at low temperature with treatment of cellulase for the isolation of ampelopsin was developed in this study.

  14. Ethnographic Contributions to Method Development

    DEFF Research Database (Denmark)

    Leander, Anna

    2016-01-01

    of IR—Critical Security Studies. Ethnographic research works with what has been termed a “strong” understanding of objectivity. When this understanding is taken seriously, it must lead to a refashioning of the processes of gathering, analyzing, and presenting data in ways that reverse many standard...... of research in the ethnographic tradition. However, it would also require rethinking standard methods instructions and the judgments they inform....... assumptions and instructions pertaining to “sound methods.” Both in the context of observation and in that of justification, working with “strong objectivity” requires a flexibility and willingness to shift research strategies that is at odds with the usual emphasis on stringency, consistency, and carefully...

  15. Developing Scoring Algorithms (Earlier Methods)

    Science.gov (United States)

    We developed scoring procedures to convert screener responses to estimates of individual dietary intake for fruits and vegetables, dairy, added sugars, whole grains, fiber, and calcium using the What We Eat in America 24-hour dietary recall data from the 2003-2006 NHANES.

  16. Taxonomic Dimensions for Studying Situational Method Development

    NARCIS (Netherlands)

    Aydin, Mehmet N.; Harmsen, Frank; van Hillegersberg, Jos; Ralyté, Jolita; Brinkkemper, Sjaak; Henderson-Sellers, Brian

    2007-01-01

    This paper is concerned with fragmented literature on situational method development, which is one of fundamental topics related to information systems development (ISD) methods. As the topic has attracted many scholars from various and possibly complementary schools of thought, different

  17. Evaluation of four methods for estimating leaf area of isolated trees

    Science.gov (United States)

    P.J. Peper; E.G. McPherson

    2003-01-01

    The accurate modeling of the physiological and functional processes of urban forests requires information on the leaf area of urban tree species. Several non-destructive, indirect leaf area sampling methods have shown good performance for homogenous canopies. These methods have not been evaluated for use in urban settings where trees are typically isolated and...

  18. Non liquid nitrogen-based-method for isolation of DNA from ...

    African Journals Online (AJOL)

    A simple, efficient, reliable and cost-effective method for isolation of total genomic DNA from fungi, suitable for polymerase chain reaction (PCR) amplification and other molecular applications was described. The main advantages of the method are: (1) does not require the use of liquid nitrogen for preparation of fungi DNA; ...

  19. Method Engineering: Engineering of Information Systems Development Methods and Tools

    OpenAIRE

    Brinkkemper, J.N.; Brinkkemper, Sjaak

    1996-01-01

    This paper proposes the term method engineering for the research field of the construction of information systems development methods and tools. Some research issues in method engineering are identified. One major research topic in method engineering is discussed in depth: situational methods, i.e. the configuration of a project approach that is tuned to the project at hand. A language and support tool for the engineering of situational methods are discussed.

  20. Antifungal susceptibility testing of vaginal candida isolates: the broth microdilution method

    Directory of Open Access Journals (Sweden)

    Mahmoudi Rad M

    2010-02-01

    Full Text Available "n Normal 0 false false false EN-US X-NONE AR-SA MicrosoftInternetExplorer4 /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0in 5.4pt 0in 5.4pt; mso-para-margin:0in; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:"Times New Roman"; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-bidi-font-family:Arial; mso-bidi-theme-font:minor-bidi;} Background: Vulvovaginal candidiasis is a common mucosal infection among immunocompetent, healthy women, and is caused by opportunistic yeasts that belong to genus Candida. In this study, we isolated and identified the Candida species in the vagina of patients who admitted in Gynecology Department of Mahdieh Hospital in Tehran, Iran to evaluate the in vitro activities of fluconazole, miconazole, itraconazole and flucytosine against 191 clinical Candida isolates by the NCCLS microdilution method."n"nMethods: 191 Candida were isolated from vaginal secretions and identified with conventional mycological methods in the diagnosis of Candida species. The identity of all strains was confirmed genotypically by multiplex PCR. In vitro susceptibility testing of vaginal Candida isolates was performed by the NCCLS broth microdilution method. The results were read at 48 h."n"nResults: Most C. albicans isolates (>90% were sensitive in vitro to the antifungal agents tested. Most C. glabrata isolates showed sensitivity to miconazole and then flucytosine while they were more resistant to Itraconazole and fluconazole. Many isolates of C. tropicalis were susceptible to miconazole and then fluconazole. They showed a little resistance to

  1. Method for isolation and molecular characterization of extracellular microvesicles released from brain endothelial cells

    Directory of Open Access Journals (Sweden)

    Haqqani Arsalan S

    2013-01-01

    Full Text Available Abstract Background In addition to possessing intracellular vesicles, eukaryotic cells also produce extracellular microvesicles, ranging from 50 to 1000 nm in diameter that are released or shed into the microenvironment under physiological and pathological conditions. These membranous extracellular organelles include both exosomes (originating from internal vesicles of endosomes and ectosomes (originating from direct budding/shedding of plasma membranes. Extracellular microvesicles contain cell-specific collections of proteins, glycoproteins, lipids, nucleic acids and other molecules. These vesicles play important roles in intercellular communication by acting as carrier for essential cell-specific information to target cells. Endothelial cells in the brain form the blood–brain barrier, a specialized interface between the blood and the brain that tightly controls traffic of nutrients and macromolecules between two compartments and interacts closely with other cells forming the neurovascular unit. Therefore, brain endothelial cell extracellular microvesicles could potentially play important roles in ‘externalizing’ brain-specific biomarkers into the blood stream during pathological conditions, in transcytosis of blood-borne molecules into the brain, and in cell-cell communication within the neurovascular unit. Methods To study cell-specific molecular make-up and functions of brain endothelial cell exosomes, methods for isolation of extracellular microvesicles using mass spectrometry-compatible protocols and the characterization of their signature profiles using mass spectrometry -based proteomics were developed. Results A total of 1179 proteins were identified in the isolated extracellular microvesicles from brain endothelial cells. The microvesicles were validated by identification of almost 60 known markers, including Alix, TSG101 and the tetraspanin proteins CD81 and CD9. The surface proteins on isolated microvesicles could potentially

  2. Developments in geophysical exploration methods

    CERN Document Server

    1982-01-01

    One of the themes in current geophysical development is the bringing together of the results of observations made on the surface and those made in the subsurface. Several benefits result from this association. The detailed geological knowledge obtained in the subsurface can be extrapolated for short distances with more confidence when the geologi­ cal detail has been related to well-integrated subsurface and surface geophysical data. This is of value when assessing the characteristics of a partially developed petroleum reservoir. Interpretation of geophysical data is generally improved by the experience of seeing the surface and subsurface geophysical expression of a known geological configuration. On the theoretical side, the understanding of the geophysical processes themselves is furthered by the study of the phenomena in depth. As an example, the study of the progress of seismic wave trains downwards and upwards within the earth has proved most instructive. This set of original papers deals with some of ...

  3. Nuclear methods in national development

    International Nuclear Information System (INIS)

    1993-01-01

    This volume of the proceedings of the First National Conference on Nuclear Methods held at Kongo Conference Hotel Zaria from 2-4 September 1993, contains the full text of about 30 technical papers and speeches of invited dignitaries presented at the conference. The technical papers are original or review articles containing results and experiences in nuclear and related analytical techniques. Topics treated include neutron generator operation and control, nuclear data, application of nuclear techniques in environment, geochemistry, medicine, biology, agriculture, material science and industries. General topics in nuclear laboratory organization and research experiences were also covered. The papers were fully discussed during the conference and authors were requested to make changes in the manuscripts where necessary. However, they were further edited. The organizing committee wishes to thank all authors for their presentation and cooperation in submitting their manuscripts promptly and the participants for their excellent contribution during the conference

  4. Comparison of the Histopaque-1119 method with the Plasmagel method for separation of blood leukocytes for cytomegalovirus isolation.

    Science.gov (United States)

    Slifkin, M; Cumbie, R

    1992-10-01

    Histopaque-1119 (Sigma Chemical Co., St. Louis, Mo.) and Plasmagel (Cellular Products, Inc., Buffalo, N.Y.) were compared as density gradient separation reagents for the separation of polymorphonuclear leukocytes and mononuclear cells from blood from the isolation of cytomegalovirus (CMV). Of 200 peripheral blood specimens examined, CMV was recovered from 51 by both methods. The time of detection of immunofluorescent sites or a cytopathic effect associated with CMV was similar by each method. The Histopaque-1119 method was less time-consuming than the Plasmagel method since it did not require a precentrifugation step for the settling of erythrocytes. The use of Histopaque-1119 will permit an effective alternative single-step method for the separation of blood leukocytes for the isolation of CMV.

  5. Remote vacuum or pressure sealing device and method for critical isolated systems

    Science.gov (United States)

    Brock, James David [Newport News, VA; Keith, Christopher D [Newport News, VA

    2012-07-10

    A remote vacuum or pressure sealing apparatus and method for making a radiation tolerant, remotely prepared seal that maintains a vacuum or pressure tight seal throughout a wide temperature range. The remote sealing apparatus includes a fixed threaded sealing surface on an isolated system, a gasket, and an insert consisting of a plug with a protruding sample holder. An insert coupling device, provided for inserting samples within the isolated system, includes a threaded fastener for cooperating with the fixed threaded sealing surface on the isolated system. The insert coupling device includes a locating pin for azimuthal orientation, coupling pins, a tooted coaxial socket wrench, and an insert coupling actuator for actuating the coupling pins. The remote aspect of the sealing apparatus maintains the isolation of the system from the user's environment, safely preserving the user and the system from detrimental effect from each respectively.

  6. Environmental friendly method for the extraction of coir fibre and isolation of nanofibre.

    Science.gov (United States)

    Abraham, Eldho; Deepa, B; Pothen, L A; Cintil, J; Thomas, S; John, M J; Anandjiwala, R; Narine, S S

    2013-02-15

    The objective of this work was to develop an environmental friendly method for the effective utilization of coir fibre by adopting steam pre-treatment. The retting of the coconut bunch makes strong environmental problems which can be avoided by this method. Chemical characterization of the fibre during each processing stages confirmed the increase of cellulose content from raw (40%) to final steam treated fibres (93%). Morphological and dynamic light scattering analyses of the fibres at different processing stages revealed that the isolation of cellulose nano fibres occur in the final step of the process as an aqueous suspension. FT-IR and XRD analysis demonstrated that the treatments lead to the gradual removal of lignin and hemicelluloses from the fibres. The existence of strong lignin-cellulose complex in the raw coir fibre is proved by its enhanced thermal stability. Steam explosion has been proved to be a green method to expand the application areas of coir fibre. Copyright © 2012 Elsevier Ltd. All rights reserved.

  7. Recent developments at the ISOL facility of GSI Darmstadt

    CERN Document Server

    Roeckl, E; Burkard, K; Döring, J; Grawe, H; Hüller, W; Kirchner, R; Mazzocchi, C; Mukha, I; Plettner, C

    2003-01-01

    The research programme pursued at the ISOL facility of GSI Darmstadt focuses on the study of decay properties of nuclei along the N congruent with Z line between the double shell closures at sup 5 sup 6 Ni and sup 1 sup 0 sup 0 Sn. In this report, the major scientific achievements obtained in the past two years are reviewed, with particular emphasis being put on the detection techniques.

  8. Accuracy of phenotypic methicillin susceptibility methods in the detection of Staphylococcus aureus isolates carrying different SCCmec types

    Directory of Open Access Journals (Sweden)

    Pricilla DM de Matos

    2010-11-01

    Full Text Available A total of 138 isolates, 118 methicillin-resistant Staphylococcus aureus (MRSA isolates (staphylococcal cassette chromosome type II, 20 isolates, type III, 39 isolates and type IV, 59 isolates and 20 methicillin-sensitive S. aureus isolates were evaluated by phenotypic methods: cefoxitin and oxacillin disk diffusion (DD, agar dilution (AD, latex agglutination (LA, oxacillin agar screening (OAS and chromogenic agar detection. All methods showed 100% specificity, but only the DD tests presented 100% sensitivity. The sensitivity of the other tests ranged from 82.2% (OAS-98.3% (AD. The LA test showed the second lowest sensitivity (86.4%. The DD test showed high accuracy in the detection of MRSA isolates, but there was low precision in the detection of type IV isolates by the other tests, indicating that the genotypic characteristics of the isolates should be considered.

  9. Accuracy of phenotypic methicillin susceptibility methods in the detection of Staphylococcus aureus isolates carrying different SCCmec types.

    Science.gov (United States)

    Matos, Pricilla D M de; Schuenck, Ricardo P; Cavalcante, Fernanda S; Caboclo, Roberta M F; Santos, Kátia Regina N dos

    2010-11-01

    A total of 138 isolates, 118 methicillin-resistant Staphylococcus aureus (MRSA) isolates (staphylococcal cassette chromosome type II, 20 isolates, type III, 39 isolates and type IV, 59 isolates) and 20 methicillin-sensitive S. aureus isolates were evaluated by phenotypic methods: cefoxitin and oxacillin disk diffusion (DD), agar dilution (AD), latex agglutination (LA), oxacillin agar screening (OAS) and chromogenic agar detection. All methods showed 100% specificity, but only the DD tests presented 100% sensitivity. The sensitivity of the other tests ranged from 82.2% (OAS)-98.3% (AD). The LA test showed the second lowest sensitivity (86.4%). The DD test showed high accuracy in the detection of MRSA isolates, but there was low precision in the detection of type IV isolates by the other tests, indicating that the genotypic characteristics of the isolates should be considered.

  10. Modified method for combined DNA and RNA isolation from peanut and other oil seeds.

    Science.gov (United States)

    Dang, Phat M; Chen, Charles Y

    2013-02-01

    Isolation of good quality RNA and DNA from seeds is difficult due to high levels of polysaccharides, polyphenols, and lipids that can degrade or co-precipitate with nucleic acids. Standard RNA extraction methods utilizing guanidinium-phenol-chloroform extraction has not shown to be successful. RNA isolation from plant seeds is a prerequisite for many seed specific gene expression studies and DNA is necessary in marker-assisted selection and other genetic studies. We describe a modified method to isolate both RNA and DNA from the same seed tissue and have been successful with several oil seeds including peanut, soybean, sunflower, canola, and oil radish. An additional LiCl precipitation step was added to isolate both RNA and DNA from the same seed tissues. High quality nucleic acids were observed based on A(260)/A(280) and A(260)/A(230) ratios above 2.0 and distinct bands on gel-electrophoresis. RNA was shown to be suitable for reverse transcriptase polymerase chain reaction based on actin or 60S ribosomal primer amplification and DNA was shown to have a single band on gel-electrophoresis analysis. This result shows that RNA and DNA isolated using this method can be appropriate for molecular studies in peanut and other oil containing seeds.

  11. Development and Test of Methods for Fault Detection and Isolation

    DEFF Research Database (Denmark)

    Jørgensen, R.B.

    Almost all industrial systemns are automated to ensure optimal production both in relation to energy consumtion and safety to equipment and humans. All working parts are individually subject to faults. This can lead to unacceptable economic loss or injury to people. This thesis deals with a monit......Almost all industrial systemns are automated to ensure optimal production both in relation to energy consumtion and safety to equipment and humans. All working parts are individually subject to faults. This can lead to unacceptable economic loss or injury to people. This thesis deals...

  12. Comparison of the Histopaque-1119 method with the Plasmagel method for separation of blood leukocytes for cytomegalovirus isolation.

    OpenAIRE

    Slifkin, M; Cumbie, R

    1992-01-01

    Histopaque-1119 (Sigma Chemical Co., St. Louis, Mo.) and Plasmagel (Cellular Products, Inc., Buffalo, N.Y.) were compared as density gradient separation reagents for the separation of polymorphonuclear leukocytes and mononuclear cells from blood from the isolation of cytomegalovirus (CMV). Of 200 peripheral blood specimens examined, CMV was recovered from 51 by both methods. The time of detection of immunofluorescent sites or a cytopathic effect associated with CMV was similar by each method....

  13. Reactor physics methods development at Westinghouse

    International Nuclear Information System (INIS)

    Mueller, E.; Mayhue, L.; Zhang, B.

    2007-01-01

    The current state of reactor physics methods development at Westinghouse is discussed. The focus is on the methods that have been or are under development within the NEXUS project which was launched a few years ago. The aim of this project is to merge and modernize the methods employed in the PWR and BWR steady-state reactor physics codes of Westinghouse. (author)

  14. Earth science developments in support of waste isolation

    International Nuclear Information System (INIS)

    Duguid, J.O.

    1981-01-01

    Earth science issues in geologic waste isolation can be subdivided into smaller questions that are resolvable. This approach provides a mechanism for focusing research on topics of definable priority and monitoring progress through the status of issue resolution. The status of resolution of major issues in borehole sealing, interpretation of groundwater hydrology, geochemistry, and repository performance assessment is presented. The Waste Terminal Storage Program has reached a point where the selection of sites, underground testing, and emplacement of waste can proceed on a well-defined schedule

  15. Two methods for isolating the lung area of a CT scan for density information

    International Nuclear Information System (INIS)

    Hedlund, L.W.; Anderson, R.F.; Goulding, P.L.; Beck, J.W.; Effmann, E.L.; Putman, C.E.

    1982-01-01

    Extracting density information from irregularly shaped tissue areas of CT scans requires automated methods when many scans are involved. We describe two computer methods that automatically isolate the lung area of a CT scan. Each starts from a single, operator specified point in the lung. The first method follows the steep density gradient boundary between lung and adjacent tissues; this tracking method is useful for estimating the overall density and total area of lung in a scan because all pixels within the lung area are available for statistical sampling. The second method finds all contiguous pixels of lung that are within the CT number range of air to water and are not a part of strong density gradient edges; this method is useful for estimating density and area of the lung parenchyma. Structures within the lung area that are surrounded by strong density gradient edges, such as large blood vessels, airways and nodules, are excluded from the lung sample while lung areas with diffuse borders, such as an area of mild or moderate edema, are retained. Both methods were tested on scans from an animal model of pulmonary edema and were found to be effective in isolating normal and diseased lungs. These methods are also suitable for isolating other organ areas of CT scans that are bounded by density gradient edges

  16. Investigation of colistin sensitivity via three different methods in Acinetobacter baumannii isolates with multiple antibiotic resistance.

    Science.gov (United States)

    Sinirtaş, Melda; Akalin, Halis; Gedikoğlu, Suna

    2009-09-01

    In recent years there has been an increase in life-threatening infections caused by Acinetobacter baumannii with multiple antibiotic resistance, which has lead to the use of polymyxins, especially colistin, being reconsidered. The aim of this study was to investigate the colistin sensitivity of A. baumannii isolates with multiple antibiotic resistance via different methods, and to evaluate the disk diffusion method for colistin against multi-resistant Acinetobacter isolates, in comparison to the E-test and Phoenix system. The study was carried out on 100 strains of A. baumannii (colonization or infection) isolated from the microbiological samples of different patients followed in the clinics and intensive care units of Uludağ University Medical School between the years 2004 and 2005. Strains were identified and characterized for their antibiotic sensitivity by Phoenix system (Becton Dickinson, Sparks, MD, USA). In all studied A. baumannii strains, susceptibility to colistin was determined to be 100% with the disk diffusion, E-test, and broth microdilution methods. Results of the E-test and broth microdilution method, which are accepted as reference methods, were found to be 100% consistent with the results of the disk diffusion tests; no very major or major error was identified upon comparison of the tests. The sensitivity and the positive predictive value of the disk diffusion method were found to be 100%. Colistin resistance in A. baumannii was not detected in our region, and disk diffusion method results are in accordance with those of E-test and broth microdilution methods.

  17. IoT System Development Methods

    NARCIS (Netherlands)

    Giray, G.; Tekinerdogan, B.; Tüzün, E.

    2018-01-01

    It is generally believed that the application of methods plays an important role in developing quality systems. A development method is mainly necessary for structuring the process in producing largescale and complex systems that involve high costs. Similar to the development of other systems, it is

  18. Carbapenem inactivation: a very affordable and highly specific method for phenotypic detection of carbapenemase-producing Pseudomonas aeruginosa isolates compared with other methods.

    Science.gov (United States)

    Akhi, Mohammad Taghi; Khalili, Younes; Ghotaslou, Reza; Kafil, Hossein Samadi; Yousefi, Saber; Nagili, Behroz; Goli, Hamid Reza

    2017-06-01

    This investigation was undertaken to compare phenotypic and molecular methods for detection of carbapenemase-producing Pseudomonas aeruginosa. A total of 245 non-duplicated isolates of P. aeruginosa were collected from hospitalized patients. Disc diffusion method was used to identify carbapenem-resistant bacteria. Three phenotypic methods, including Modified Hodge Test (MHT), Modified Carba NP (MCNP) test and Carbapenem Inactivation Method (CIM) were used for investigation of carbapenemase production. In addition, polymerase chain reaction (PCR) was used to detect carbapenemase encoding genes. Of 245 P. aeruginosa isolates investigated, 121 isolates were carbapenem-resistant. Among carbapenem-resistant isolates, 40, 39 and 35 isolates exhibited positive results using MHT, MCNP test and CIM, respectively. PCR indicated the presence of carbapenemase genes in 35 of carbapenem-resistant isolates. MHT showed low sensitivity and specificity for carbapenemase detection among P. aeruginosa isolates in comparison to PCR. CIM was most affordable and highly specific than MCNP test compared with the molecular method.

  19. Enrichment Method for the Isolation of Bioactive Actinomycetes From Mangrove Sediments of Andaman Islands, India

    Directory of Open Access Journals (Sweden)

    Baskaran, R.

    2011-01-01

    Full Text Available Various pre-treatment methods and three different media were employed for the isolation of bioactive actinomycetes from mangrove sediments of Andaman and Nicobar Islands, India. Sediments from four different sites of mangrove forest were collected and pre-treated by dry heat method, and the media were supplemented with cycloheximide 80 µg/mL and nalidixic acid 75 µg/mL. The mean actinomycetes population density in sediment samples were recorded as 22 CFU-10^-6/gm in KUA medium followed by 12 CFU-10^-6/gm in AIA medium and 8 CFU-10^-6/gm in SCA medium. A total of 42 actinomycetes were isolated, and all the isolates were evaluated for their antibacterial activity against pathogenic bacteria on two different media. Among 42 isolates tested, 22 species were found to be antibacterial metabolite producer against test bacteria namely, Staphylococcus aureus, Bacillus subtilis, Salmonella typhi and Klebsiella pneumoniae. Particularly, the actinomycete strains such as A101, A102, A107, A116, A121, A125, A130, F101, F102, F104, F106, De101 and De102 significantly inhibited the growth of all bacteria which were tested. Of these strains, A107 was identified as Streptomyces spp. This strain had the maximum activity against all used pathogens on both medium. Hence, the isolation, characterization and studies of secondary metabolites of actinomycetes from mangrove sediments in Andaman and Nicobar Island could be a pathway for discovery of antibiotics from marine actinomycetes.

  20. Hanford Permanent Isolation Barrier Program: Asphalt technology development

    International Nuclear Information System (INIS)

    Freeman, H.D.; Romine, R.A.

    1994-11-01

    An important component of the Hanford Permanent Isolation Barrier is the use of a two-layer composite asphalt system, which provides backup water diversion capabilities if the primary capillary barrier fails to meet infiltration goals. Because of asphalt's potential to perform to specification over the 1000-year design life criterion, a composite asphalt barrier (HMAC/fluid-applied polymer-modified asphalt) is being considered as an alternative to the bentonite clay/high density poly(ethylene) barriers for the low-permeability component of the Hanford Permanent Isolation Barrier. The feasibility of using asphalt as a long-term barrier is currently being studied. Information that must be known is the ability of asphalt to retain desirable physical properties over a period of 1000 years. This paper presents the approach for performing accelerated aging tests and evaluating the performance of samples under accelerated conditions. The results of these tests will be compared with asphalt artifact analogs and the results of modeling the degradation of the selected asphalt composite to make life-cycle predictions

  1. A safe inexpensive method to isolate high quality plant and fungal ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-08-18

    Aug 18, 2008 ... quality DNA from plant and fungal species. This method uses potassium acetate to remove proteins and polysaccharides in an SDS extraction buffer. Further DNA purification is achieved using a low salt. CTAB treatment. This SDS/CTAB protocol was used to isolate high quality genomic DNA subject to.

  2. Methods for Isolation, Purification, and Propagation of Bacteriophages of Campylobacter jejuni

    DEFF Research Database (Denmark)

    Gencay, Yilmaz Emre; Birk, Tina; Sørensen, Martine Camilla Holst

    2017-01-01

    Here, we describe the methods for isolation, purification, and propagation of Campylobacter jejuni bacteriophages from samples expected to contain high number of phages such as chicken feces. The overall steps are (1) liberation of phages from the sample material; (2) observation of plaque-formin...

  3. Three methods for isolating viable anthozoan endoderm cells with their intracellular symbiotic dinoflagellates

    Science.gov (United States)

    Gates, R. D.; Muscatine, L.

    1992-09-01

    Three maceration methods are described for the isolation of single endoderm cells from marine cnidarians. Two are enzymatic treatments suitable for fleshy anthozoans such as sea anemones and zoanthids. The third employs calcium free sea water and is suitable for stony corals. The viability and morphology of the endoderm cells is described using fluorogenic dyes and scanning and transmission electron microscopy.

  4. Development of Dissolution Test Method for Drotaverine ...

    African Journals Online (AJOL)

    Development of Dissolution Test Method for Drotaverine ... Methods: Sink conditions, drug stability and specificity in different dissolution media were tested to optimize a dissolution test .... test by Prism 4.0 software, and differences between ...

  5. A Modified Ficoll-Paque Gradient Method for Isolating Mononuclear Cells from the Peripheral and Umbilical Cord Blood of Humans for Biobanks and Clinical Laboratories.

    Science.gov (United States)

    Jia, Yanjuan; Xu, Hui; Li, Yonghong; Wei, Chaojun; Guo, Rui; Wang, Fang; Wu, Yu; Liu, Jing; Jia, Jing; Yan, Junwen; Qi, Xiaoming; Li, Yuanting; Gao, Xiaoling

    2018-04-01

    Although the Ficoll-Paque method is classically used to isolate peripheral blood mononuclear cells (PBMCs), modifications in this method are required for a more rapid and economic output for biobanks and clinical laboratories, particularly in developing countries. In this study, we addressed this issue by modifying the Ficoll-Paque method for the isolation of PBMCs or mononuclear cells from the peripheral and the umbilical cord blood of healthy and diseased (infected, anemic, and chronic obstructive pulmonary disease) adult individuals. In the modified method, we initiated the cell isolation process from the buffy coat layer, which appears in the interface between the plasma and sediments after centrifugation, instead of using the whole blood as described in the classic method. Although the PBMC yield by the modified method was about 12% less than in the classic method, the number of PBMCs isolated by the modified method was more than one million, which is enough for different research/diagnostic purposes, such as multi-omics detection. Assessment of cell viability and purity by hematology analyzer and trypan blue showed no significant difference between the viability and purity of the PBMCs isolated by these two methods in almost all groups, except samples from the infected and cord blood groups, where lower PBMC purity with higher granulocyte contamination were observed. In addition, at delayed processing time points, all parameters for the two methods were decreased in a time-dependent manner, especially at 8, 12, or 24 hours after the sample collection. In summary, the performance of PBMC isolation by the classic and modified methods mainly relies on the PBMC ratio in original samples. The modified method could be preferred for PBMC isolation because of its time and cost savings, especially for the biobanks and clinical laboratories in developing countries.

  6. A novel tertiary prep-HPLC method for the isolation of single amino acids for AMS-radiocarbon measurement.

    Science.gov (United States)

    Fernandes, Ricardo; Koudelka, Tomas; Tholey, Andreas; Dreves, Alexander

    2017-07-15

    AMS-radiocarbon measurements of amino acids can potentially provide more reliable radiocarbon dates than bulk collagen analysis. Nonetheless, the applicability of such an approach is often limited by the low-throughput of existing isolation methods and difficulties in determining the contamination introduced during the separation process. A novel tertiary prep-HPLC amino acid isolation method was developed that relies on the combustion of eluted material without requiring any additional chemical steps. Amino acid separation was carried out using a gradient mix of pure water and phosphoric acid with an acetonitrile step in-between runs to remove hydrophobic molecules from the separation column. The amount of contaminant carbon and its 14 C content were determined from two-point measurements of collagen samples of known 14 C content. The amount of foreign carbon due to the isolation process was estimated at 4±1μg and its 14 C content was 0.43±0.01 F 14 C. Radiocarbon values corrected for carbon contamination have only a minor increase in uncertainties. For Holocene samples, this corresponds to an added uncertainty typically smaller than 10 14 Cyears. The developed method can be added to routine AMS measurements without implying significant operational changes and offers a level of measurement uncertainty that is suitable for many archaeological, ecological, environmental, and biological applications. Copyright © 2017. Published by Elsevier B.V.

  7. A simple cleanup method for the isolation of nitrate from natural water samples for O isotopes analysis

    International Nuclear Information System (INIS)

    Haberhauer, G.; Blochberger, K.

    1999-09-01

    The analysis of O-isotopic composition of nitrate has many potential applications in studies of environmental processes. O-isotope nitrate analysis require sample free of other oxygen-containing compounds. More than 100 % of non-NO 3 - oxygen relative to NO 3 - oxygen can still be found in forest soil water samples after cleanup if improper cleanup strategies, e.g., adsorption onto activated carbon, are used. Such non-NO 3 - oxygen compounds will bias O-isotropic data. Therefore, an efficient cleanup method was developed to isolate nitrate from natural water samples. In a multistep cleanup procedure using adsorption onto water-insoluble poly(vinylpyrrolidone), removal of almost all other oxygen-containing compounds, such as fulvic acids, and isolation of nitrate was achieved. The method supplied samples free of non-NO 3 - oxygen which can be directly combusted to CO 2 for subsequent O-isotope analysis. (author)

  8. Moral counselling: a method in development.

    Science.gov (United States)

    de Groot, Jack; Leget, Carlo

    2011-01-01

    This article describes a method of moral counselling developed in the Radboud University Medical Centre Nijmegen (The Netherlands). The authors apply insights of Paul Ricoeur to the non-directive counselling method of Carl Rogers in their work of coaching patients with moral problems in health care. The developed method was shared with other health care professionals in a training course. Experiences in the course and further practice led to further improvement of the method.

  9. Development, characterization and application of monoclonal antibodies against Brazilian Dengue virus isolates.

    Directory of Open Access Journals (Sweden)

    Camila Zanluca

    Full Text Available Dengue is the most prevalent human arboviral disease. The morbidity related to dengue infection supports the need for an early, quick and effective diagnostic test. Brazil is a hotspot for dengue, but no serological diagnostic test has been produced using Brazilian dengue virus isolates. This study aims to improve the development of immunodiagnostic methods for dengue virus (DENV detection through the production and characterization of 22 monoclonal antibodies (mAbs against Brazilian isolates of DENV-1, -2 and -3. The mAbs include IgG2bκ, IgG2aκ and IgG1κ isotypes, and most were raised against the envelope or the pre-membrane proteins of DENV. When the antibodies were tested against the four DENV serotypes, different reactivity patterns were identified: group-specific, subcomplex specific (DENV-1, -3 and -4 and DENV-2 and -3 and dengue serotype-specific (DENV-2 or -3. Additionally, some mAbs cross-reacted with yellow fever virus (YFV, West Nile virus (WNV and Saint Louis encephalitis virus (SLEV. None of the mAbs recognized the alphavirus Venezuelan equine encephalitis virus (VEEV. Furthermore, mAbs D3 424/8G, D1 606/A12/B9 and D1 695/12C/2H were used to develop a capture enzyme-linked immunosorbent assay (ELISA for anti-dengue IgM detection in sera from patients with acute dengue. To our knowledge, these are the first monoclonal antibodies raised against Brazilian DENV isolates, and they may be of special interest in the development of diagnostic assays, as well as for basic research.

  10. Methods to isolate a large amount of generative cells, sperm cells and vegetative nuclei from tomato pollen for omics analysis

    Directory of Open Access Journals (Sweden)

    Yunlong eLu

    2015-06-01

    Full Text Available The development of sperm cells from microspores involves a set of finely regulated molecular and cellular events and the coordination of these events. The mechanisms underlying these events and their interconnections remain a major challenge. Systems analysis of genome-wide molecular networks and functional modules with high-throughput omics approaches is crucial for understanding the mechanisms; however, this study is hindered because of the difficulty in isolating a large amount of cells of different types, especially generative cells (GCs, from the pollen. Here, we optimized the conditions of tomato pollen germination and pollen tube growth to allow for long-term growth of pollen tubes in vitro with sperm cells (SCs generated in the tube. Using this culture system, we developed methods for isolating GCs, SCs and vegetative-cell nuclei (VN from just-germinated tomato pollen grains and growing pollen tubes and their purification by Percoll density gradient centrifugation. The purity and viability of isolated GCs and SCs were confirmed by microscopy examination and fluorescein diacetate staining, respectively, and the integrity of VN was confirmed by propidium iodide staining. We could obtain about 1.5 million GCs and 2.0 million SCs each from 180 mg initiated pollen grains, and 10 million VN from 270 mg initiated pollen grains germinated in vitro in each experiment. These methods provide the necessary preconditions for systematic biology studies of SC development and differentiation in higher plants.

  11. Methods to isolate a large amount of generative cells, sperm cells and vegetative nuclei from tomato pollen for "omics" analysis.

    Science.gov (United States)

    Lu, Yunlong; Wei, Liqin; Wang, Tai

    2015-01-01

    The development of sperm cells (SCs) from microspores involves a set of finely regulated molecular and cellular events and the coordination of these events. The mechanisms underlying these events and their interconnections remain a major challenge. Systems analysis of genome-wide molecular networks and functional modules with high-throughput "omics" approaches is crucial for understanding the mechanisms; however, this study is hindered because of the difficulty in isolating a large amount of cells of different types, especially generative cells (GCs), from the pollen. Here, we optimized the conditions of tomato pollen germination and pollen tube growth to allow for long-term growth of pollen tubes in vitro with SCs generated in the tube. Using this culture system, we developed methods for isolating GCs, SCs and vegetative cell nuclei (VN) from just-germinated tomato pollen grains and growing pollen tubes and their purification by Percoll density gradient centrifugation. The purity and viability of isolated GCs and SCs were confirmed by microscopy examination and fluorescein diacetate staining, respectively, and the integrity of VN was confirmed by propidium iodide staining. We could obtain about 1.5 million GCs and 2.0 million SCs each from 180 mg initiated pollen grains, and 10 million VN from 270 mg initiated pollen grains germinated in vitro in each experiment. These methods provide the necessary preconditions for systematic biology studies of SC development and differentiation in higher plants.

  12. An Overview of Neolignans of the Genus Piper L.: Isolation Methods and Biological Activities.

    Science.gov (United States)

    Macedo, Arthur Ladeira; Dos Santos, Thais Carvalho Costa; Valverde, Alessandra Leda; Moreira, Davyson de Lima; Vasconcelos, Thatyana Rocha Alves

    2017-01-01

    The genus Piper L. has the shikimic acid pathway predominantly expressed, biosynthesizing many cinnamic acid derivatives (CAD). Neolignans comprise an important class of CAD that exhibit a wide range of pharmacological properties such as antibacterial, antitumor, insecticidal, anti-inflammatory, antioxidant, smooth muscle relaxant, neuroprotective, antiprotozoal and against platelet aggregation factor. These substances have been extracted and isolated from Piper species using different technics. The present review aims to summarize extraction and isolation methods and biological activities of the different types of neolignans covering the period from 1968 to January 2016. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  13. COMPARATIVE EVALUATION OF CONVENTIONAL VERSUS RAPID METHODS FOR AMPLIFIABLE GENOMIC DNA ISOLATION OF CULTURED Azospirillum sp. JG3

    Directory of Open Access Journals (Sweden)

    Stalis Norma Ethica

    2013-12-01

    Full Text Available As an initial attempt to reveal genetic information of Azospirillum sp. JG3 strain, which is still absence despite of the strains' ability in producing valued enzymes, two groups of conventional methods: lysis-enzyme and column-kit; and two rapid methods: thermal disruption and intact colony were evaluated. The aim is to determine the most practical method for obtaining high-grade PCR product using degenerate primers as part of routine-basis protocols for studying the molecular genetics of the Azospirillal bacteria. The evaluation includes the assessment of electrophoresis gel visualization, pellet appearance, preparation time, and PCR result of extracted genomic DNA from each method. Our results confirmed that the conventional methods were more superior to the rapid methods in generating genomic DNA isolates visible on electrophoresis gel. However, modification made in the previously developed DNA isolation protocol giving the simplest and most rapid method of all methods used in this study for extracting PCR-amplifiable DNA of Azospirillum sp. JG3. Intact bacterial cells (intact colony loaded on electrophoresis gel could present genomic DNA band, but could not be completely amplified by PCR without thermal treatment. It can also be inferred from our result that the 3 to 5-min heating in dH2O step is critical for the pre-treatment of colony PCR of Azospirillal cells.

  14. Impact of enumeration method on diversity of Escherichia coli genotypes isolated from surface water.

    Science.gov (United States)

    Martin, E C; Gentry, T J

    2016-11-01

    There are numerous regulatory-approved Escherichia coli enumeration methods, but it is not known whether differences in media composition and incubation conditions impact the diversity of E. coli populations detected by these methods. A study was conducted to determine if three standard water quality assessments, Colilert ® , USEPA Method 1603, (modified mTEC) and USEPA Method 1604 (MI), detect different populations of E. coli. Samples were collected from six watersheds and analysed using the three enumeration approaches followed by E. coli isolation and genotyping. Results indicated that the three methods generally produced similar enumeration data across the sites, although there were some differences on a site-by-site basis. The Colilert ® method consistently generated the least diverse collection of E. coli genotypes as compared to modified mTEC and MI, with those two methods being roughly equal to each other. Although the three media assessed in this study were designed to enumerate E. coli, the differences in the media composition, incubation temperature, and growth platform appear to have a strong selective influence on the populations of E. coli isolated. This study suggests that standardized methods of enumeration and isolation may be warranted if researchers intend to obtain individual E. coli isolates for further characterization. This study characterized the impact of three USEPA-approved Escherichia coli enumeration methods on observed E. coli population diversity in surface water samples. Results indicated that these methods produced similar E. coli enumeration data but were more variable in the diversity of E. coli genotypes observed. Although the three methods enumerate the same species, differences in media composition, growth platform, and incubation temperature likely contribute to the selection of different cultivable populations of E. coli, and thus caution should be used when implementing these methods interchangeably for

  15. A development of three-dimensional seismic isolation for advanced reactor systems in Japan: Pt.2

    International Nuclear Information System (INIS)

    Kenji Takahashi; Kazuhiko Inoue; Asao Kato; Masaki Morishita; Takafumi Fujita

    2005-01-01

    Two types of three-dimensional seismic isolation systems were developed for the fast breeder reactor (FBR). One is the three-dimensional entire building base isolation system It was developed by collecting concepts Japanese companies from which a combination system with air springs and hydraulic rocking suppression devices was selected. The other is the vertically isolated system for main components with horizontally entire building base isolation, which was developed by adopting coned disk spring devices. In the study, seismic condition was assumed based on a strict reference ground motion. Design data of the building and components are referred to FBR being developed as the 'Commercialized Fast Reactor Cycle System'. Analysis based on these assumed conditions showed suitable combinations of natural frequencies and damping ratios for isolation. Devices were developed to satisfy the combinations. In five years research and development, several verification tests were performed including shake table tests with scaled models. Finally it is found that the two types of seismic isolation systems are available for FBR. The result is reflected in the preliminary design guideline for the three-dimensional isolation system. (authors)

  16. An improved culture method for selective isolation of Campylobacter jejuni from wastewater

    Directory of Open Access Journals (Sweden)

    Jinyong Kim

    2016-08-01

    Full Text Available Campylobacter jejuni is one of the leading foodborne pathogens worldwide. C. jejuni is isolated from a wide range of foods, domestic animals, wildlife, and environmental sources. The currently-available culture-based isolation methods are not highly effective for wastewater samples due to the low number of C. jejuni in the midst of competing bacteria. To detect and isolate C. jejuni from wastewater samples, in this study, we evaluated a few different enrichment conditions using five different antibiotics (i.e., cefoperazone, vancomycin, trimethoprim, polymyxin B, and rifampicin, to which C. jejuni is intrinsically resistant. The selectivity of each enrichment condition was measured with Ct value using quantitative real-time PCR (qRT-PCR, and multiplex PCR to determine Campylobacter species. In addition, the efficacy of Campylobacter isolation on different culture media after selective enrichment was examined by growing on Bolton and Preston agar plates. The addition of polymyxin B, rifampicin, or both to the Bolton selective supplements enhanced the selective isolation of C. jejuni. In particular, rifampicin supplementation and an increased culture temperature (i.e., 42°C had a decisive effect on the selective enrichment of C. jejuni from wastewater. The results of 16S rDNA sequencing also revealed that Enterococcus spp. and Pseudomonas aeruginosa are major competing bacteria in the enrichment conditions. Although it is known to be difficult to isolate Campylobacter from samples with heavy contamination, this study well exhibited that the manipulation of antibiotic selective pressure improves the isolation efficiency of fastidious Campylobacter from wastewater.

  17. Comparison of different DNA isolation methods and use of dodecyle trimethyl ammonium bromide (DTAB for the isolation of DNA from meat products

    Directory of Open Access Journals (Sweden)

    Yusuf OZsENSOY

    2016-12-01

    Conclusion: DNA isolation kit, another best method, is recommended due to quality and quantity of DNA for researchers who do not want that phenol/chloroform method have toxic substances. This study is also the first study in which DTAB method is used for DNA extraction from meat products. [J Adv Vet Anim Res 2016; 3(4.000: 368-374

  18. Methodology for developing new test methods

    Directory of Open Access Journals (Sweden)

    A. I. Korobko

    2017-06-01

    Full Text Available The paper describes the methodology for developing new test methods and forming solutions for the development of new test methods. The basis of the methodology for developing new test methods is the individual elements of the system and process approaches. They contribute to the development of an effective research strategy for the object, the study of interrelations, the synthesis of an adequate model of the test method. The effectiveness of the developed test method is determined by the correct choice of the set of concepts, their interrelations and mutual influence. This allows you to solve the tasks assigned to achieve the goal. The methodology is based on the use of fuzzy cognitive maps. The question of the choice of the method on the basis of which the model for the formation of solutions is based is considered. The methodology provides for recording a model for a new test method in the form of a finite set of objects. These objects are significant for the test method characteristics. Then a causal relationship is established between the objects. Further, the values of fitness indicators and the observability of the method and metrological tolerance for the indicator are established. The work is aimed at the overall goal of ensuring the quality of tests by improving the methodology for developing the test method.

  19. A highly efficient nonchemical method for isolating live nematodes (Caenorhabditis elegans) from soil during toxicity assays.

    Science.gov (United States)

    Kim, Shin Woong; Moon, Jongmin; An, Youn-Joo

    2015-01-01

    The success of soil toxicity tests using Caenorhabditis elegans may depend in large part on recovering the organisms from the soil. However, it can be difficult to learn the International Organization for Standardization/ASTM International recovery process that uses the colloidal silica flotation method. The present study determined that a soil-agar isolation method provides a highly efficient and less technically demanding alternative to the colloidal silica flotation method. Test soil containing C. elegans was arranged on an agar plate in a donut shape, a linear shape, or a C curve; and microbial food was placed outside the soil to encourage the nematodes to leave the soil. The effects of ventilation and the presence of food on nematode recovery were tested to determine the optimal conditions for recovery. A linear arrangement of soil on an agar plate that was sprinkled with microbial food produced nearly 83% and 90% recovery of live nematodes over a 3-h and a 24-h period, respectively, without subjecting the nematodes to chemical stress. The method was tested using copper (II) chloride dihydrate, and the resulting recovery rate was comparable to that obtained using colloidal silica flotation. The soil-agar isolation method portrayed in the present study enables live nematodes to be isolated with minimal additional physicochemical stress, making it a valuable option for use in subsequent sublethal tests where live nematodes are required. © 2014 SETAC.

  20. The SIESTA method; developments and applicability

    International Nuclear Information System (INIS)

    Artacho, Emilio; Anglada, E; Dieguez, O; Gale, J D; Garcia, A; Junquera, J; Martin, R M; Ordejon, P; Pruneda, J M; Sanchez-Portal, D; Soler, J M

    2008-01-01

    Recent developments in and around the SIESTA method of first-principles simulation of condensed matter are described and reviewed, with emphasis on (i) the applicability of the method for large and varied systems (ii) efficient basis sets for the standards of accuracy of density-functional methods (iii) new implementations, and (iv) extensions beyond ground-state calculations

  1. Some problems on Monte Carlo method development

    International Nuclear Information System (INIS)

    Pei Lucheng

    1992-01-01

    This is a short paper on some problems of Monte Carlo method development. The content consists of deep-penetration problems, unbounded estimate problems, limitation of Mdtropolis' method, dependency problem in Metropolis' method, random error interference problems and random equations, intellectualisation and vectorization problems of general software

  2. An intercomparison of three methods for the large-scale isolation of oceanic dissolved organic matter

    Science.gov (United States)

    Green, Nelson W.; Perdue, E. Michael; Aiken, George R.; Butler, Kenna D.; Chen, Hongmei; Dittmar, Thorsten; Niggemann, Jutta; Stubbins, Aron

    2014-01-01

    Dissolved organic matter (DOM) was isolated from large volumes of deep (674 m) and surface (21 m) ocean water via reverse osmosis/electrodialysis (RO/ED) and two solid-phase extraction (SPE) methods (XAD-8/4 and PPL) at the Natural Energy Laboratory of Hawaii Authority (NELHA). By applying the three methods to common water samples, the efficiencies of XAD, PPL and RO/ED DOM isolation were compared. XAD recovered 42% of dissolved organic carbon (DOC) from deep water (25% with XAD-8; 17% with XAD-4) and 30% from surface water (16% with XAD-8; 14% with XAD-4). PPL recovered 61 ± 3% of DOC from deep water and 61% from surface water. RO/ED recovered 82 ± 3% of DOC from deep water, 14 ± 3% of which was recovered in a sodium hydroxide rinse, and 75 ± 5% of DOC from surface water, with 12 ± 2% in the sodium hydroxide rinse. The highest recoveries of all were achieved by the sequential isolation of DOC, first with PPL and then via RO/ED. This combined technique recovered 98% of DOC from a deep water sample and 101% of DOC from a surface water sample. In total, 1.9, 10.3 and 1.6 g-C of DOC were collected via XAD, PPL and RO/ED, respectively. Rates of DOC recovery using the XAD, PPL and RO/ED methods were 10, 33 and 10 mg-C h− 1, respectively. Based upon C/N ratios, XAD isolates were heavily C-enriched compared with water column DOM, whereas RO/ED and PPL ➔ RO/ED isolate C/N values were most representative of the original DOM. All techniques are suitable for the isolation of large amounts of DOM with purities suitable for most advanced analytical techniques. Coupling PPL and RO/ED techniques may provide substantial progress in the search for a method to quantitatively isolate oceanic DOC, bringing the entirety of the DOM pool within the marine chemist's analytical window.

  3. Biodiversity of soil-mycoflora of islamabad pakistan using two isolation methods

    International Nuclear Information System (INIS)

    Anjum, F.; Asif, M.; Ayub, N.

    2005-01-01

    The soil microfungal flora of Islamabad were investigated, by using the direct plate and dilution plate methods. A total of 4,225 fungal colonies were isolated from 80 samples collected from Islamabad, Pakistan. The species-composition revealed 66 different species belonging to 20 fungal genera. Among these, Penicillium, being most common and prevalent, was ranked as the dominant genus with respect to the test-sites. Most of the species belong to genera Aspergillus and Penicillium. The maximum number of colonies i.e. 1797 were developed on DRBC, followed by PDA (1320) and the minimum number was recorded on SDA (1108). The results indicate that 4 of these species belong to Mucorales, one to Sphaeriales, one to Coelomycetes and 60 to Hyphomycetes. The most widespread genera were Aspergillus (16 species), Penicillium (15 species) and Acremonium (7 species). The most common species were Aspergillus niger (241 colonies), Cladosporium cladosporioides (219 colonies), Penicillium charlesii (201 colonies), Aspergillus flavus (200 colonies), Penicillium frequantus (198 colonies), Penicillium purpurogenum (178 colonies) and Alternaria alternata (176 colonies). Direct plating and dilution plating with three media, i.e. Potato Dextrose Agar, Dichloran Rose Bengal Agar, and Sabrouad Dextrose Agar, were used in this study. Dilution plating with Dichloran Rose Bengal Chloramphenicol Agar (DRBC) was found to be the most suitable. Other two media i.e. Potato Dextrose Agar (PDA) and Sabrouad Dextrose Agar (SDA) were also tried for cultivation of fungal species with successful results. (author)

  4. A screening method for the isolation of polyhydroxyalkanoate-producing purple non-sulfur photosynthetic bacteria from natural seawater

    Directory of Open Access Journals (Sweden)

    Mieko Higuchi-Takeuchi

    2016-09-01

    Full Text Available Polyhydroxyalkanoates (PHAs are a family of biopolyesters accumulated by a variety of microorganisms as carbon and energy storage under starvation conditions. We focused on marine purple non-sulfur photosynthetic bacteria as host microorganisms for PHA production and developed a method for their isolation from natural seawater. To identify novel PHA-producing marine purple non-sulfur photosynthetic bacteria, natural seawaters were cultured in nutrient-rich medium for purple non-sulfur photosynthetic bacteria, and twelve pink- or red-pigmented colonies were picked up. Gas chromatography mass spectrometry analysis revealed that four isolates synthesized PHA at levels ranging from 0.5 to 24.4 wt% of cell dry weight. The 16S ribosomal RNA sequence analysis revealed that one isolate (HM2 showed 100% identity to marine purple non-sulfur photosynthetic bacteria. In conclusion, we have demonstrated in this study that PHA-producing marine purple non-sulfur photosynthetic bacteria can be isolated from natural seawater under nutrient-rich conditions.

  5. Development of additional pituitary hormone deficiencies in pediatric patients originally diagnosed with idiopathic isolated GH deficiency

    NARCIS (Netherlands)

    W.F. Blum (Werner); C.L. Deal (Cheri Lynn); A.G. Zimmermann (Alan); E.P. Shavrikova (Elena); C.J. Child (Christopher); C.A. Quigley (Charmian); S.L.S. Drop (Stenvert); G. Cutler (Gordon); R.G. Rosenfeld (Ron)

    2014-01-01

    textabstractObjective: We assessed the characteristics of children initially diagnosed with idiopathic isolated GH deficiency (IGHD) who later developed additional (multiple) pituitary hormone deficiencies (MPHD). Design: Data were analyzed for 5805 pediatric patients with idiopathic IGHD, who were

  6. Development of novel agar media for isolating guaiacol producing Alicyclobacillus spp.

    Science.gov (United States)

    Chang, S S; Park, S H; Kang, D H

    2013-06-03

    The purpose of this study is to develop a selective and differential medium (SK2 agar) for isolating guaiacol producing Alicyclobacillus. Forty-one selected dyes and vanillic acid were incorporated in SK agar for screening selective and differential agents. Two guaiacol producing (1016, 1101) and two non-guaiacol producing (19220, C-GD 1-1) Alicyclobacillus isolates were streaked onto media and color differentiation of the isolates was assessed. Among 41 tested dyes, Chrome Azurol S (CAS) allowed color differentiation of the two types of Alicyclobacillus. Colonies of guaiacol producing Alicyclobacillus isolates appeared as dark purple to royal blue color with yellow background, whereas non-guaiacol producing Alicyclobacillus isolates produced cream colored colonies with yellow background. Vanillic acid not only served as a precursor for guaiacol formation but also inhibited non-guaiacol producing Alicyclobacillus. Non-guaiacol producing isolates did not grow on SK agar containing more than 70 ppm vanillic acid, whereas the recovery of guaiacol producing isolates was unaffected. When compared with other Alicyclobacillus isolation media, not only was SK2 agar capable of selectively recovering guaiacol-producing Alicyclobacillus, the degree of growth was also approximately equal if not better than orange serum agar, potato dextrose agar, and K agar. The development of SK2 agar provides the fruit juice industry with an inexpensive, simple to use alternative for the detection of guaiacol producing Alicyclobacillus. Copyright © 2013 Elsevier B.V. All rights reserved.

  7. The Improved Method for Isolation of Photochrome Trans-membrane Protein Bacteriorhodopsin from Purple Membranes of Halobacterium Halobacterium Halobium ET 1001

    OpenAIRE

    Oleg Mosin; Ignat Ignatov

    2015-01-01

    It was developed the improved method for isolation of photochrome trans-membraine protein bacteriorhodopsin (output – 5 mg from 100 g of wet biomass) capable to transform light energy to electrochemical energy of generated protons H+ and АТP. The protein was isolated from purple membranes of photo-organotrophic halobacterium Halobacterium halobium ET 1001 by cellular autolysis by distilled water, processing of bacterial biomass by ultrasound at 22 KHz, alcohol extraction of low and high-weigh...

  8. Fine typing of methicillin-resistant Staphylococcus aureus isolates using direct repeat unit and staphylococcal interspersed repeat unit typing methods.

    Science.gov (United States)

    Ho, Cheng-Mao; Ho, Mao-Wang; Li, Chi-Yuan; Lu, Jang-Jih

    2015-08-01

    Methicillin-resistant Staphylococcus aureus (MRSA) typing is an important epidemiologic tool for monitoring trends and preventing outbreaks. However, the efficiency of various MRSA typing methods for each SCCmec MRSA isolate is rarely evaluated. A total of 157 MRSA isolates from four different regions in Taiwan were typed with five different molecular methods, including SCCmec typing, multilocus sequence typing (MLST), spa typing, mec-associated direct repeat unit (dru) copy number determination, and staphylococcal interspersed repeat unit (SIRU) profiling. There were four SCCmec types, eight MLST types, 15 spa types, 11 dru types, and 31 SIRU profiles. The most common type determined by each molecular typing method was SCCmec III (115 isolates, 73.2%), ST239 (99 isolates, 63.1%), t037 (107 isolates, 68.2%), 14 dru copies (76 isolates, 48.4%), and SIRU profile 3013722 (102 isolates, 65%), respectively. When using the combination of MLST, spa typing, and dru copy number, ST5-t002-4 (n = 8), ST239-t037-14 (n = 68), ST59-t437-9 (n = 9), and ST59-t437-11 (n = 6) were found to be the most common types of SCCmec types II (n = 9), III (n = 115), IV (n = 21), and VT (n = 11) isolates, respectively. SCCmec type III isolates were further classified into 11 dru types. Of the 21 SCCmec type IV isolates, 14 SIRU profiles were found. Seven SIRU patterns were observed in the 11 SCCmec type VT isolates. Different typing methods showed a similar Hunter-Gaston discrimination index among the 157 MRSA isolates. However, dru and SIRU typing methods had a better discriminatory power for SCCmec type III and SCCmec types IV and VT isolates, respectively, suggesting that dru and SIRU can be used to further type these isolates. Copyright © 2013. Published by Elsevier B.V.

  9. Isolation of developing secondary xylem specific cellulose synthase ...

    Indian Academy of Sciences (India)

    The present study aimed at identifying developing secondary xylem specific cellulose synthase genes from .... the First strand cDNA synthesis kit (Fermentas, Pittsburgh,. USA). .... ing height of the rooted cutting, girth of the stem, leaf area.

  10. Development of a Thermal Isolation Structure for Aerospace Cryogenic Instruments

    Science.gov (United States)

    Nash, A.; Robeck, L.

    1999-01-01

    A proof of concept prototype cryostat has been developed to demonstrate the ability to accommodate low temperature science investigations within the constraints of the Hitchhiker siderail carrier on the space shuttle.

  11. Comparative studies of two methods for miRNA isolation from milk whey.

    Science.gov (United States)

    Jin, Xiao-lu; Wei, Zi-hai; Liu, Lan; Liu, Hong-yun; Liu, Jian-xin

    2015-06-01

    MicroRNAs (miRNAs) from milk whey have been considered for their potential as noninvasive biomarkers for milk quality control and disease diagnosis. However, standard protocols for miRNA isolation and quantification from milk whey are not well established. The objective of this study was to compare two methods for the isolation of miRNAs from milk whey. These two methods were modified phenol-based technique (Trizol LS(®) followed by phenol precipitation, the TP method) and combined phenol and column-based approach (Trizol LS(®) followed by cleanup using the miRNeasy kit, the TM method). Yield and quality of RNA were rigorously measured using a NanoDrop ND-1000 spectrophotometer and then the distribution of RNA was precisely detected in a Bioanalyzer 2100 instrument by microchip gel electrophoresis. Several endogenous miRNAs (bta-miR-141, bta-miR-146a, bta-miR-148a, bta-miR-200c, bta-miR-362, and bta-miR-375) and an exogenous spike-in synthetic control miRNA (cel-miR-39) were quantified by real-time polymerase chain reaction (PCR) to examine the apparent recovery efficiency of milk whey miRNAs. Both methods could successfully isolate sufficient small RNA (whey, and their yields were quite similar. However, the quantification results show that the total miRNA recovery efficiency by the TM method is superior to that by the TP method. The TM method performed better than the TP for recovery of milk whey miRNA due to its consistency and good repeatability in endogenous and spike-in miRNA recovery. Additionally, quantitative recovery analysis of a spike-in miRNA may be more accurate to reflect the milk whey miRNA recovery efficiency than using traditional RNA quality analysis instruments (NanoDrop or Bioanalyzer 2100).

  12. Comparative studies of two methods for miRNA isolation from milk whey*

    Science.gov (United States)

    Jin, Xiao-lu; Wei, Zi-hai; Liu, Lan; Liu, Hong-yun; Liu, Jian-xin

    2015-01-01

    MicroRNAs (miRNAs) from milk whey have been considered for their potential as noninvasive biomarkers for milk quality control and disease diagnosis. However, standard protocols for miRNA isolation and quantification from milk whey are not well established. The objective of this study was to compare two methods for the isolation of miRNAs from milk whey. These two methods were modified phenol-based technique (Trizol LS® followed by phenol precipitation, the TP method) and combined phenol and column-based approach (Trizol LS® followed by cleanup using the miRNeasy kit, the TM method). Yield and quality of RNA were rigorously measured using a NanoDrop ND-1000 spectrophotometer and then the distribution of RNA was precisely detected in a Bioanalyzer 2100 instrument by microchip gel electrophoresis. Several endogenous miRNAs (bta-miR-141, bta-miR-146a, bta-miR-148a, bta-miR-200c, bta-miR-362, and bta-miR-375) and an exogenous spike-in synthetic control miRNA (cel-miR-39) were quantified by real-time polymerase chain reaction (PCR) to examine the apparent recovery efficiency of milk whey miRNAs. Both methods could successfully isolate sufficient small RNA (whey, and their yields were quite similar. However, the quantification results show that the total miRNA recovery efficiency by the TM method is superior to that by the TP method. The TM method performed better than the TP for recovery of milk whey miRNA due to its consistency and good repeatability in endogenous and spike-in miRNA recovery. Additionally, quantitative recovery analysis of a spike-in miRNA may be more accurate to reflect the milk whey miRNA recovery efficiency than using traditional RNA quality analysis instruments (NanoDrop or Bioanalyzer 2100). PMID:26055915

  13. [Means and methods of personal hygiene in the experiment with 520-day isolation].

    Science.gov (United States)

    Shumilina, G A; Shumilina, I V; Solov'eva, S O

    2013-01-01

    Six volunteers (3 Russians, a Frenchman, an Italian and a Chinese) participated in assessment of the input of sanitation and housekeeping provisions to their wellbeing during 520-day isolation and confinement. Subject of the study was quality and sufficiency of housekeeping agents and procedures as well as more than 60 names of personal hygiene items. The sanitation and housekeeping monitoring involved the clinical, hygienic and microbiological methods, and also consideration of crew comments on the items at their disposal and recommended procedures. Based on the analysis of the functional condition of the integument and oral cavity and entries in the questionnaires, i.e. objective data and subjective feelings, all test subjects remained in the invariably good state. Owing to the application of the selected hygienic means and methods the microbial status of the crew was stable throughout 520-day isolation.

  14. Comparison of Plasmagel with LeucoPREP-Macrodex methods for separation of leukocytes for virus isolation.

    Science.gov (United States)

    Woods, G L; Proffitt, M R

    1987-10-01

    Plasmagel (Cellular Products, Inc., Buffalo, NY), which can separate both polymorphonuclear leukocytes (PMN) and mononuclear cells from other blood components, and LeucoPREP (Becton Dickinson Immunocytometry Systems, Mountain View, CA), which can separate mononuclear cells from other blood components, were used to harvest leukocytes from whole blood for the purpose of virus isolation. Macrodex was combined with the later, in a second step, for recovery of PMN. Of 90 peripheral blood specimens examined, cytomegalovirus was recovered from 10: in six by both methods, in three from Plasmagel prepared cells only, and in one from cells from the LeucoPREP-Macrodex preparation only. Total leukocyte counts, differential counts, and leukocyte viability did not differ significantly for the two methods. Plasmagel provided an efficient, inexpensive means of harvesting leukocytes from whole blood for virus isolation.

  15. Isolation, Screening and Development of Local Bacterial Consortia With Azo Dyes Decolourising Capability

    Directory of Open Access Journals (Sweden)

    Khadijah, O.

    2009-01-01

    Full Text Available A total of 1540 bacterial isolates were isolated and screened for their ability to degrade selected azo dyes. Of these, nine isolates were chosen for further studies based on their ability to degrade a wide spectrum of dyes efficiently and rapidly. Several microbial consortia were developed and tested for their effectiveness. Overall the consortia were able to degrade 70 - 100% colour within 72 hours compared to 60 – 97% colour removed by individual isolates. A microbial consortium labelled C15 showed good growth in agitation culture but the colour removal was best in static culture with 80 - 100% colour removed in less than 72 hours. Based on the 16S rRNA sequencing, two of the bacterial isolates in C15 belong to the Chryseobacterium genus while the other one belongs to Flavobacterium genus.

  16. Prognostic aspects of imaging method development

    International Nuclear Information System (INIS)

    Steinhart, L.

    1987-01-01

    A survey is presented of X-ray diagnostic methods and techniques and possibilities of their further development. Promising methods include direct imaging using digital radiography. In connection with computer technology these methods achieve higher resolution. The storage of obtained images in the computer memory will allow automated processing and evaluation and the use of expert systems. Development is expected to take place especially in computerized tomography using magnetic resonance, and positron computed tomography and other non-radioactive diagnostic methods. (J.B.). 5 figs., 1 tab., 1 ref

  17. Improved Method for the Isolation of Biosurfactant Glycolipids from Rhodococcus sp. Strain H13A

    OpenAIRE

    Bryant, Frank O.

    1990-01-01

    An improved method for the isolation of the biosurfactant glycolipids from Rhodococcus sp. strain H13A by using XM 50 diafiltration and isopropanol precipitation was devised. This procedure was advantageous since it removes protein coisolated when the glycolipids are obtained by organic extraction and silicic acid chromatography. The protein apparently does not contribute any biosurfactant characteristics to the glycolipids. The deacylated glycolipid backbone included only a disaccharide.

  18. Sensitive microculture method for isolation of human immunodeficiency virus type 1 from blood leukocytes.

    Science.gov (United States)

    Erice, A; Sannerud, K J; Leske, V L; Aeppli, D; Balfour, H H

    1992-02-01

    A study was conducted to compare our standard culture with a new microculture procedure for isolation of human immunodeficiency virus type 1 (HIV-1) from blood leukocytes. A total of 137 blood specimens from 102 HIV-1 antibody-positive individuals (52 were asymptomatic, 31 were symptomatic, and 19 had AIDS) were cultured in a microculture system in which 10(6) of the patients' peripheral blood mononuclear cells (PBMC) were cocultured with 10(6) phytohemagglutinin (PHA)-stimulated PBMC from an HIV-1 antibody-negative blood donor in 1.2 ml of culture medium. Results were compared with those of a historical control group of 139 standard HIV-1 cultures from 108 HIV-1 antibody-positive subjects (58 were asymptomatic, 36 were symptomatic, and 14 had AIDS). For standard cultures, 10 x 10(6) of the patients' PBMC were cocultured with 5 x 10(6) PHA-stimulated PBMC from an HIV-1 antibody-negative blood donor in 15 ml of culture medium. HIV-1 was isolated in 128 (93%) microcultures and 133 (96%) standard cultures. Both methods identified more than 75% of the positive cultures within 7 days and 100% of the positive cultures within 14 days. The isolation rates for HIV-1 in microcultures compared with standard cultures were 91 versus 93% (specimens from asymptomatic individuals), 93 versus 96% (specimens from symptomatic individuals), and 97 versus 100% (specimens from patients with AIDS). The median time to positivity for both culture methods was 7 days, and this correlated significantly with symptoms and CD4+ cell counts. The microculture method is a sensitive and less expensive system for isolation of HIV-1 from PBMC of HIV-1 antibody-positive individuals, and we recommend it as the culture method of choice, especially for children and patients with AIDS and severe anemia or leukopenia whose blood volume is an important consideration.

  19. Development of Antioxidant Activity during Milk Fermentation by Wild Isolates of Lactobacillus helveticus

    Directory of Open Access Journals (Sweden)

    Aazam Namdari

    2016-07-01

    Full Text Available Background and Objective: Oxidative stress, due to free radicals, brings injury to the body by attacking large molecules and cell organs, and is the main reason of many diseases. Fermentation of foods containing large amount of proteins such as milk by special species of lactic acid bacteria is a potential way in enhancement of the antioxidative activity of foods. This study aimed at evaluating non-common starter species isolates of Lactobacillus helveticus for their capability to produce fermented milk enriched in antioxidant peptides.Materials and Methods: Reconstituted skim milk (11% was inoculated with 7 wild isolates of Lactobacillus helveticus, and after 24 h fermentation at 37ºC, the samples were kept 4ºC and for 14 days. Viable cell number, acidification and proteolysis degree in the milk fermented by each isolate were assessed in 1, 7 and 14 days. Development of antioxidant activity was measured using DPPH and ABTS●+ radial scavenging activities during the storage period.Results and Conclusion: Though some slight strain-dependent differences were observed in growth, acidification and proteolysis, all the samples showed considerably strong antioxidant activity (at least 62.32±3.66% and 57.64±1.42% measured using DPPH and ABTS●+ radicals, respectively through the whole storage period. In vitro simulated gastrointestinal digestion indicated that DPPH radical-scavenging activity of the antioxidative peptidic supernatants was not affected significantly by consecutive pepsin-pancreatin hydrolysis in most of the samples. These evidences support Lactobacillus helveticus as a promising functional culture able to promote health benefits in dairy-based functional foods.Conflict of interest: The authors declare that there is no conflict of interest.

  20. Contribution of the JRC Ispra to the intercomparison of analysis methods for seismically isolated nuclear structures

    International Nuclear Information System (INIS)

    Magonette, G.; Renda, V.

    2002-01-01

    Aim of the work done at JRC has been essentially to investigate the potentiality of the Pseudo-Dynamic (PsD) method to test structures incorporating anti-seismic protection devices based on materials with a strain-rate dependent behaviour. This is of relevant importance due to the interest to perform tests on large-scale mock-ups to assess the behaviour of realistic structure of civil engineering interest. Two specific typologies of protection have been analysed and tested at the European Laboratory for Structural Assessment (ELSA) of JRC Ispra. The first dealing with base isolation and the second with energy dissipation devices. In both cases the protection devices were based on high damping rubber material which is characterised by a moderate dependence from the strain rate of the application of the displacements. To validate a standard procedure to test base isolated structures by the PsD method, a collaboration was set up with the Italian Working Group on Seismic Isolation which includes the national research centre ENEA, the national electricity board ENEL, the industrial research centre ISMES and a manufacturer of isolators ALGA. In the framework of this collaboration it was decided to test at the ELSA laboratory a scaled 5-storey frame structure (provided by ENEL), isolated by means of high damping rubber bearings (HDRBs), which had been tested on the shaking table of ISMES. This experimental activity aimed to compare the results which can be obtained by means of the PsD testing technique with those which can be obtained by means of a truly-dynamic test on a shaking table. To validate a standard procedure to test structures incorporating energy dissipation devices, an international collaboration has been set up with Industries, Research Centres and Universities in the framework of a project partially funded by the European Commission through the General Directorate for Science and Technology. The obtained results show once more that the PsD method, when

  1. Energy and economic development in isolated regions: a case study

    International Nuclear Information System (INIS)

    Munoz, A.; Maldonado, P.

    1992-01-01

    High production costs and inadequate supply of electricity are severe handicaps to economic development in remote areas. Costs are increased further by the low level and high variability of electric loads. In order to face these problems a simultaneous development of wind diesel energy resources and productive projects with demand management is proposed. This paper illustrates such a case in Southern Chile where present electricity costs are reduced as consumption is increased from 85 to 468 KWh/person/year. A new refrigeration plant increases the storage capacity for the small fishing activity and effectively uses the output of wind energy generators. (author)

  2. Development of Antibiotic Resistance Against Ureaplasma urealyticum Strains Isolated from Urogenital Samples

    Directory of Open Access Journals (Sweden)

    Musa Saraçoğlu

    2018-04-01

    Full Text Available Objective: To assess any change in the antibiotic sensitivity of Ureaplasma urealyticum strains isolated from urogenital samples in the course of time. Materials and Methods: Hospital records were retrospectively examined and cases with growth of U. urealyticum in urogenital samples in the years 2008 and 2013 were identified. Furthermore, the change in the course of time was examined by taking into consideration the cases we reported in 2001. Results: Higher rates of sensitivity against tetracycline and doxycycline were observed in 60 patients with isolated U. urealyticum. Higher rates of resistance against ofloxacin and ciprofloxacin were observed. A significant difference was found in resistance against antibiotics when the records of 2008 and 2013 were compared. A statistically significant increase was found in resistance against ofloxacin and ciprofloxacin when the records of 2001 were compared with the records of 2008 and 2013 (p<0.0005. Conclusion: U. urealyticum strains demonstrated high levels of resistance to quinolones. Resistance development is increasing in the course of time. Sensitivity against tetracycline and doxycycline has continued at high rates. It would be beneficial to consider these results during empirical treatment to be applied in cases ineligible for culturing.

  3. A fast and reproducible method for albumin isolation and depletion from serum and cerebrospinal fluid.

    Science.gov (United States)

    Holewinski, Ronald J; Jin, Zhicheng; Powell, Matthew J; Maust, Matthew D; Van Eyk, Jennifer E

    2013-03-01

    Analysis of serum and plasma proteomes is a common approach for biomarker discovery, and the removal of high-abundant proteins, such as albumin and immunoglobins, is usually the first step in the analysis. However, albumin binds peptides and proteins, which raises concerns as to how the removal of albumin could impact the outcome of the biomarker study while ignoring the possibility that this could be a biomarker subproteome itself. The first goal of this study was to test a new commercially available affinity capture reagent from Protea Biosciences and to compare the efficiency and reproducibility to four other commercially available albumin depletion methods. The second goal of this study was to determine if there is a highly efficient albumin depletion/isolation system that minimizes sample handling and would be suitable for large numbers of samples. Two of the methods tested (Sigma and ProteaPrep) showed an albumin depletion efficiency of 97% or greater for both serum and cerebrospinal fluid (CSF). Isolated serum and CSF albuminomes from ProteaPrep spin columns were analyzed directly by LC-MS/MS, identifying 128 serum (45 not previously reported) and 94 CSF albuminome proteins (17 unique to the CSF albuminome). Serum albuminome was also isolated using Vivapure anti-HSA columns for comparison, identifying 105 proteins, 81 of which overlapped with the ProteaPrep method. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  4. Development of a new method to identify aminoacylated RNA

    Directory of Open Access Journals (Sweden)

    Wang Ji

    2014-02-01

    Full Text Available A RT-PCR method is developed to isolate RNA aminoacylated on their 3’ end from large pools of RNA. The method is being applied in two separate projects. We are interested in isolating a new class of ribozymes that could successively catalyze the two chemical reactions leading to their own 3’ aminoacylation (ATP activation of an amino acid followed by 3' esterification of the RNA. The catalysis of each of the two reactions has independently been demonstrated for some RNA isolated with the SELEX methodology [1-2]. However, the coupling of both reactions on a same molecule has not been achieved yet. The identification of these still hypothetical ribozymes may help understand how the former translation system started in the absence of the aminoacyltRNA Synthetase, which catalyzes the above two reactions on tRNA in modern cells. In another project, we would like to identify the whole repertoire of aminoacylated RNA (the “aminoacylome” in cells. There are strong indications that other RNA besides tRNA and tmRNA may be aminoacylated for biological purposes [3-4].

  5. Development of a variable stiffness and damping tunable vibration isolator

    CSIR Research Space (South Africa)

    Cronje, JM

    2005-03-01

    Full Text Available the amplification at resonance. A practical variable stiffness spring was developed by using a compound leaf spring with circular spring elements. A wax actuator, controlled by a hot-air gun with a closed-loop displacement and velocity feedback control system...

  6. Development of spectrophotometric fingerprinting method for ...

    African Journals Online (AJOL)

    Selective and efficient analytical methods are required not only for quality assurance but also for authentication of herbal formulations. A simple, rapid and validated fingerprint method has developed for estimation of piperine in 'Talisadi churna', a well known herbal formulation in India. The estimation was carried out in two ...

  7. Validated modified Lycopodium spore method development for ...

    African Journals Online (AJOL)

    Validated modified lycopodium spore method has been developed for simple and rapid quantification of herbal powdered drugs. Lycopodium spore method was performed on ingredients of Shatavaryadi churna, an ayurvedic formulation used as immunomodulator, galactagogue, aphrodisiac and rejuvenator. Estimation of ...

  8. Development of Three Methods for Simultaneous Quantitative ...

    African Journals Online (AJOL)

    Development of Three Methods for Simultaneous Quantitative Determination of Chlorpheniramine Maleate and Dexamethasone in the Presence of Parabens in ... Tropical Journal of Pharmaceutical Research ... Results: All the proposed methods were successfully applied to the analysis of raw materials and dosage form.

  9. Development and Application of Kinetic Spectrophotometric Method ...

    African Journals Online (AJOL)

    Purpose: To develop an improved kinetic-spectrophotometric procedure for the determination of metronidazole (MNZ) in pharmaceutical formulations. Methods: The method is based on oxidation reaction of MNZ by hydrogen peroxide in the presence of Fe(II) ions at pH 4.5 (acetate buffer). The reaction was monitored ...

  10. Tracking maize pollen development by the Leaf Collar Method.

    Science.gov (United States)

    Begcy, Kevin; Dresselhaus, Thomas

    2017-12-01

    An easy and highly reproducible nondestructive method named the Leaf Collar Method is described to identify and characterize the different stages of pollen development in maize. In plants, many cellular events such as meiosis, asymmetric cell division, cell cycle regulation, cell fate determination, nucleus movement, vacuole formation, chromatin condensation and epigenetic modifications take place during pollen development. In maize, pollen development occurs in tassels that are confined within the internal stalk of the plant. Hence, identification of the different pollen developmental stages as a tool to investigate above biological processes is impossible without dissecting the entire plant. Therefore, an efficient and reproducible method is necessary to isolate homogeneous cell populations at individual stages throughout pollen development without destroying the plant. Here, we describe a method to identify the various stages of pollen development in maize. Using the Leaf Collar Method in the maize inbreed line B73, we have determined the duration of each stage from pollen mother cells before meiosis to mature tricellular pollen. Anther and tassel size as well as percentage of pollen stages were correlated with vegetative stages, which are easily recognized. The identification of stage-specific genes indicates the reproducibility of the method. In summary, we present an easy and highly reproducible nondestructive method to identify and characterize the different stages of pollen development in maize. This method now opens the way for many subsequent physiological, morphological and molecular analyses to study, for instance, transcriptomics, metabolomics, DNA methylation and chromatin patterns during normal and stressful conditions throughout pollen development in one of the economically most important grass species.

  11. An inexpensive and fast method for infiltration coating of complex geometry matrices for ISOL production target applications

    International Nuclear Information System (INIS)

    Kawai, Y.; Alton, G.D.; Bilheux, J.-C.

    2005-01-01

    An inexpensive, fast, and close to universal infiltration coating technique has been developed for fabricating fast diffusion-release ISOL targets. Targets are fabricated by deposition of finely divided (∼1μm) compound materials in a paint-slurry onto highly permeable, complex structure reticulated-vitreous-carbon-foam (RVCF) matrices, followed by thermal heat treatment. In this article, we describe the coating method and present information on the physical integrity, uniformity of deposition, and matrix adherence of SiC, HfC and UC 2 targets, destined for on-line use as targets at the Holifield Radioactive Ion Beam Facility (HRIBF)

  12. [Comparison between conventional methods, ChromAgar Candida® and PCR method for the identification of Candida species in clinical isolates].

    Science.gov (United States)

    Estrada-Barraza, Deyanira; Dávalos Martínez, Arturo; Flores-Padilla, Luis; Mendoza-De Elias, Roberto; Sánchez-Vargas, Luis Octavio

    2011-01-01

    The increase in the incidence of yeast species causing fungemia in susceptible immunocompromised patients in the last two decades and the low sensitivity of conventional blood culture has led to the need to develop alternative approaches for the early detection and identification of causative species. The aim of this study was to compare the usefulness of molecular testing by the polymerase chain reaction (PCR) and conventional methods to identify clinical isolates of different species, using the ID32C ATB system (bioMérieux, France), chromogenic culture Chromagar Candida® (CHROMagar, France) and morphogenesis in corn meal agar. We studied 79 isolates, in which the most prevalent species using the system ID32C and PCR was C. albicans, followed by C. tropicalis, C. glabrata and C .krusei. PCR patterns obtained for the identification of clinical isolates were stable and consistent in the various independent studies and showed good reproducibility, concluding that PCR with species-specific primers that amplify genes ITS1 and ITS2 for rRNA or topoisomerase II primers is a very specific and sensitive method for the identification of C. glabrata, C. krusei, C. albicans, and with less specificity for C. tropicalis. Copyright © 2010 Revista Iberoamericana de Micología. Published by Elsevier Espana. All rights reserved.

  13. An improved method for RNA isolation and cDNA library construction from immature seeds of Jatropha curcas L

    Directory of Open Access Journals (Sweden)

    Kaur Jatinder

    2010-05-01

    Full Text Available Abstract Background RNA quality and quantity is sometimes unsuitable for cDNA library construction, from plant seeds rich in oil, polysaccharides and other secondary metabolites. Seeds of jatropha (Jatropha curcas L. are rich in fatty acids/lipids, storage proteins, polysaccharides, and a number of other secondary metabolites that could either bind and/or co-precipitate with RNA, making it unsuitable for downstream applications. Existing RNA isolation methods and commercial kits often fail to deliver high-quality total RNA from immature jatropha seeds for poly(A+ RNA purification and cDNA synthesis. Findings A protocol has been developed for isolating good quality total RNA from immature jatropha seeds, whereby a combination of the CTAB based RNA extraction method and a silica column of a commercial plant RNA extraction kit is used. The extraction time was reduced from two days to about 3 hours and the RNA was suitable for poly(A+ RNA purification, cDNA synthesis, cDNA library construction, RT-PCR, and Northern hybridization. Based on sequence information from selected clones and amplified PCR product, the cDNA library seems to be a good source of full-length jatropha genes. The method was equally effective for isolating RNA from mustard and rice seeds. Conclusions This is a simple CTAB + silica column method to extract high quality RNA from oil rich immature jatropha seeds that is suitable for several downstream applications. This method takes less time for RNA extraction and is equally effective for other tissues where the quality and quantity of RNA is highly interfered by the presence of fatty acids, polysaccharides and polyphenols.

  14. Social isolation in childhood and adult inflammation: evidence from the National Child Development Study.

    Science.gov (United States)

    Lacey, Rebecca E; Kumari, Meena; Bartley, Mel

    2014-12-01

    Social isolation is known to be associated with poorer health amongst adults, including coronary heart disease. It is hypothesized that this association may be mediated by inflammation. There has been little prospective research on the long-term impact of social isolation in childhood on adult health or the pathways which might be involved. The aim of this study was to investigate whether social isolation in childhood is associated with increased adult inflammation and the mechanisms involved across the life course. This study used multiply-imputed data on 7462 participants of the National Child Development Study in Great Britain. The association between child social isolation (7-11 yrs) and levels of C-reactive protein (CRP) in middle age (44 yrs) was examined. We additionally investigated the role of adult social isolation, psychological distress, health behaviors and socioeconomic factors as potential mediators using path analysis and concurrent measurements made across the life course. Socially isolated children had higher levels of C-reactive protein in mid-life (standardized coefficient=0.05, p≤0.001). In addition, children who were socially isolated tended to have lower subsequent educational attainment, be in a less advantaged social class in adulthood, were more likely to be psychologically distressed across adulthood and were more likely to be obese and to smoke. All of these factors partially explained the association between childhood social isolation and CRP. However, this association remained statistically significant after considering all mediators simultaneously. Social isolation in childhood is associated with higher levels of C-reactive protein in mid-life. This is explained in part through complex mechanisms acting across the life course. Identification and interventions targeted toward socially isolated children may help reduce long-term adult health risk. Copyright © 2014 The Authors. Published by Elsevier Ltd.. All rights reserved.

  15. Possible approaches to community development for nuclear waste isolation

    International Nuclear Information System (INIS)

    Voth, D.E.; Herrington, B.E.

    1982-10-01

    Community development is a process whereby communities assess their needs, determine their priorities, make plans, carry out these plans, and evaluate the results. This approach is applied in evaluating four possible decision-making scenarios that DOE could use in the site selection process and in the development of compensation/mitigation mechanisms. In the first scenario, DOE would select the repository site purely on geologic and technological grounds. In the second scenario, DOE would select the repository site using geologic and technological information, detailed socioeconomic information, and an understanding of potential socioeconomic impacts of a nuclear repository upon the community. In the third scenario DOE would select a repository site using the same information available in the second scenario; however, in this case, DOE would provide the local community with professionals who would act as advocates for the local community and argue their position within DOE's decision-making process. In the final scenario, DOE would invite potential repository site communities to engage in an auctioning process, whereby each community would bid on an acceptable compensation package (in their eyes) to be paid by DOE. Based on an evaluation of these scenarios, the report makes the following recommendations: (1) DOE should consider the auctioning process; (2) in the event that no communities are willing to present bids, DOE should provide communities with advocates; (3) compensation should be a collective package rather than in the form of individual cash payments; (4) a third party (other than DOE), with a heavy reliance on local organizations, should be specifically created for implementing the compensation/mitigation package; and (5) in order to approach the siting of additional repositories with greater understanding, longitudinal research at the first repository should be undertaken as a part of the monitoring phase

  16. A method for starting up the gas isolating section of a benzine pyrolysis installation

    Energy Technology Data Exchange (ETDEWEB)

    Bruskin, Yu.A.; Gorokhov, V.V.; Kotler, L.D.; Shevchenko, K.N.; Zeldin, V.Ye.

    1982-01-01

    In the method for starting up a gas isolation section of a benzine pyrolysis unit, which includes starting the demethanizer in a steaming mode, starting the ethane and ethylene and propane and propylene towers, filling the ethylene cooling system with a hydrocarbon fraction, starting the ethylene cooling system, switching to a mode of the demethanization unit, starting the subassemblies for hydration and isolation of the ethylene and propylene fractions, in order to reduce the length of the start up period and to reduce the ejection of gas to the burner, after starting the demethanizer in the steaming mode, starting the ethane and ethylene and propane and propylene towers, the ethane and ethylene and propane and propylene fractions are mixed before the hydration subassemblies with an H2 bearing gas for catalytic reforming and then the other units and subassemblies are started.

  17. A Study on the Development of Prototype Seismic Isolation Device for NPP

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Hongpyo; Cho, Myungsug; Kim, Sunyong; Lee, Yonghee; Kang Kyunghun [KHNP-CRI, Daejeon (Korea, Republic of)

    2014-05-15

    Korean nuclear power plants have been and still are based on seismic resistance design including all of the natural disasters. However, in regions of high seismic hazard, seismic isolation technology is needed to guarantee the seismic safety on nuclear power plants. To achieve this purpose, the research and development of seismic isolation system for the construction in high seismicity area is on-going in Korea. In this study, prototype seismic isolation devices as mentioned above are developed and tested to identify the basic shear and compressive characteristics of them. In this study, assessment performance of basic characteristics on the prototype LRB and EQS seismic isolation for nuclear power plant structures is employed to compare with design values. Based on the test results of compression and shear characteristics, it is judged that they meet the measuring efficiency range conditions which are presented in ISO 22762 and AASHOT guide specification. Therefore, prototype seismic isolation devices like LRB and EQS developed in this study can be expected to be used as reference data when designing a seismic isolation system for nuclear power plant structures in the future.

  18. Phene Plate (PhP) biochemical fingerprinting. A screening method for epidemiological typing of enterococcal isolates.

    Science.gov (United States)

    Saeedi, B; Tärnberg, M; Gill, H; Hällgren, A; Jonasson, J; Nilsson, L E; Isaksson, B; Kühn, I; Hanberger, H

    2005-09-01

    Pulsed-field gel electrophoresis (PFGE) is currently considered the gold standard for genotyping of enterococci. However, PFGE is both expensive and time-consuming. The purpose of this study was to investigate whether the PhP system can be used as a reliable clinical screening method for detection of genetically related isolates of enterococci. If so, it should be possible to minimize the number of isolates subjected to PFGE typing, which would save time and money. Ninety-nine clinical enterococcal isolates were analysed by PhP (similarity levels 0.90-0.975) and PFGE (similarity levels PhP also belong to the same cluster according to PFGE, i.e. p(A(PFGE)=B(PFGE) * A(PhP)=B(PhP)), and the probability that a pair of isolates of different types according to PhP also belong to different clusters according to PFGE, i.e. p(A(PFGE) not equalB(PFGE) * A(PhP) not equalB(PhP)), was relatively high for E. faecalis (0.86 and 0.96, respectively), but was lower for E. faecium (0.51 and 0.77, respectively). The concordance which shows the probability that PhP and PFGE agree on match or mismatch was 86%-93% for E. faecalis and 54%-66% for E. faecium, which indicates that the PhP method may be useful for epidemiological typing of E. faecalis in the current settings but not for E. faecium.

  19. Lectin-induced agglutination method of urinary exosomes isolation followed by mi-RNA analysis: Application for prostate cancer diagnostic.

    Science.gov (United States)

    Samsonov, Roman; Shtam, Tatiana; Burdakov, Vladimir; Glotov, Andrey; Tsyrlina, Evgenia; Berstein, Lev; Nosov, Alexander; Evtushenko, Vladimir; Filatov, Michael; Malek, Anastasia

    2016-01-01

    low-cost and easily performed method for isolation of exosomes from urine. Isolated exosomes can be further analysed in terms of miRNA content. The miRNA profile of urinary exosomes reflects development of prostate cancer and may present a promising diagnostic tool. © 2015 Wiley Periodicals, Inc.

  20. Comparison of Methods for Isolating High Quality DNA and RNA from an Oleaginous Fungus Cunninghamella bainieri Strain 2a1

    OpenAIRE

    Noor Adila, A. K.; Farah Diba, A. B.; Zamri, Z.; Wan Mohtar, W. Y.; Aidil, A. H.; Mahadi, N. M.; Murad, A. M. A.

    2007-01-01

    A number of protocols have been reported for efficient fungal DNA and RNA isolation. However, many of these methods are often designed for certain groups or morphological forms of fungi and, in some cases, are species dependent. In this report, we compared four published protocols for DNA isolation from a locally isolated oleaginous fungus, Cunninghamella bainieri strain 2a1. These protocols either involved the use of polyvinyl pyrrolidone (PVP), hexacetyltrimethylammonium bromide (CTAB) or w...

  1. Development of antheridial filaments of Chara vulgaris L. in isolated antheridia in vitro

    Directory of Open Access Journals (Sweden)

    Hanna Kuran

    2014-01-01

    Full Text Available The study was undertaken to establish the conditions of culture in vitro of Chara vulgaris L. antheridium isolated from the mother plant and to ascertain the regularities in the development of the antheridial filaments under these conditions. The culture of isolated antheridia was run for 5 days on Forsberg medium. The antheridial filaments were found to preserve their full viability evaluated in terms of 3H-phenylalanine incorporation With simultaneous depression of mitotic activity. Under these conditions a considerable part of the filaments pass through at least three division cycles. In vitro culture of isolated antheridia causes the greater shortening of the cell length the longer the antheridia are kept Isolated from the thallus.

  2. Development of Methods of Innovative Projects’ Management in Developer Organizations

    Directory of Open Access Journals (Sweden)

    Papelniuk Oksana

    2017-01-01

    Full Text Available In article the author studies the conceptual scheme of life cycle of innovative developer project and offers methods of innovative projects' management according to the stages of its lifecycle. The author describes algorithm of the organization of management system of innovative projects in the developer organization representing set of stages and methods. The offered algorithm allows developer organizations achieve one of the main of their objectives, which is the creation of a special environment of comfortable accommodation with the emphasis on ecological compatibility and energy efficiency.

  3. Isolation and Identification of Volatile Components in Tempe by Simultaneous Distillation-Extraction Method by Modified Extraction Method

    Directory of Open Access Journals (Sweden)

    Syahrial Syahrial

    2010-06-01

    Full Text Available An isolation and identification of volatile components in temps for 2, 5 and 8 days fermentation by simultaneous distillation-extraction method was carried out. Simultaneous distillation-extraction apparatus was modified by Muchalal from the basic Likens-Nickerson's design. Steam distillation and benzena as an extraction solvent was used in this system. The isolation was continuously carried out for 3 hours which maximum water temperature In the Liebig condenser was 8 °C. The extract was concentrated by freeze concentration method, and the volatile components were analyzed and identified by combined gas chromatography-mass spectrophotometry (GC-MS. The Muchalal's simultaneous distillation extraction apparatus have some disadvantage in cold finger condenser, and it's extractor did not have condenser. At least 47, 13 and 5 volatile components were found in 2, 5 and 8 days fermentation, respectively. The volatile components in the 2 days fermentation were nonalal, ɑ-pinene, 2,4-decadienal, 5-phenyldecane, 5-phenylundecane, 4-phenylundecane, 5-phenyldodecane, 4-phenyldodecane, 3-phenyldodecane, 2-phenyldodecane, 5-phenyltridecane, and caryophyllene; in the 5 days fermentation were nonalal, caryophyllene, 4-phenylundecane, 5-phenyldodecane, 4-phenyldodecane, 3-phenyldodecane, 2-phenyldodecane; and in the 8 days fermentation were ethenyl butanoic, 2-methy1-3-(methylethenylciclohexyl etanoic and 3,7-dimethyl-5-octenyl etanoic.

  4. Methods for Confirming the Gram Reaction of Gram-variable Bacillus Species Isolated from Tobacco

    Directory of Open Access Journals (Sweden)

    Morin A

    2014-12-01

    Full Text Available Bacillus is a predominant genus of bacteria isolated from tobacco. The Gram stain is the most commonly used and most important of all diagnostic staining techniques in microbiology. In order to help confirm the Gram positivity of Bacillus isolates from tobacco, three methods using the chemical differences of the cell wall and membrane of Gram-positive and Gram-negative bacteria were investigated: the KOH (potassium hydroxide, the LANA (L-alanine-4-nitroanilide, and the vancomycin susceptibility tests. When colonies of Gram-negative bacteria are treated with 3% KOH solution, a slimy suspension is produced, probably due to destruction of the cell wall and liberation of deoxyribonucleic acid (DNA. Gram-positive cell walls resist KOH treatment. The LANA test reveals the presence of a cell wall aminopeptidase that hydrolyzes the L-alanine-4-nitroanilide in Gram-negative bacteria. This enzyme is absent in Gram-positive bacteria. Vancomycin is a glycopeptide antibiotic inhibiting the cell wall peptido-glycan synthesis of Gram-positive microorganisms. Absence of lysis with KOH, absence of hydrolysis of LANA, and susceptibility to vancomycin were used with the Gram reaction to confirm the Gram positivity of various Bacillus species isolated from tobacco. B. laevolacticus excepted, all Bacillus species tested showed negative reactions to KOH and LANA tests, and all species were susceptible to vancomycin (5 and 30 µg.

  5. Disturbances of dental development distinguish patients with oligodontia-ectodermal dysplasia from isolated oligodontia.

    Science.gov (United States)

    Dhamo, B; Kuijpers, M A R; Balk-Leurs, I; Boxum, C; Wolvius, E B; Ongkosuwito, E M

    2018-02-01

    To investigate phenotypic differences in dental development between isolated oligodontia and oligodontia-ectodermal dysplasia (ED). A total of 129 patients diagnosed with isolated oligodontia and 22 patients with oligodontia as part of ED were eligible. The phenotype of dental development was assessed for the frequency of missing a certain tooth, dental age, development of each tooth present, abnormal size and abnormal shape of teeth. The data were analysed building linear, ordinal and logistic regression models. Compared to patients with isolated oligodontia, patients with oligodontia-ED missed more frequently central incisors and second molars in both jaws, and lateral incisors in the mandible (P ectodermal abnormalities. © 2017 The Authors. Orthodontics & Craniofacial Research Published by John Wiley & Sons Ltd.

  6. Development of advanced MCR task analysis methods

    International Nuclear Information System (INIS)

    Na, J. C.; Park, J. H.; Lee, S. K.; Kim, J. K.; Kim, E. S.; Cho, S. B.; Kang, J. S.

    2008-07-01

    This report describes task analysis methodology for advanced HSI designs. Task analyses was performed by using procedure-based hierarchical task analysis and task decomposition methods. The results from the task analysis were recorded in a database. Using the TA results, we developed static prototype of advanced HSI and human factors engineering verification and validation methods for an evaluation of the prototype. In addition to the procedure-based task analysis methods, workload estimation based on the analysis of task performance time and analyses for the design of information structure and interaction structures will be necessary

  7. Effects of Phytophthora cinnamomi isolate, inoculum delivery method, flood, and drought on vigor, disease severity and mortality of blueberry plants

    Science.gov (United States)

    Four studies evaluated the effect of Phytophthora cinnamomi isolates, inoculum delivery methods, and flood and drought conditions on vigor, disease severity scores, and survival of blueberry plants grown in pots in the greenhouse. Phytophthora cinnamomi isolates were obtained from blueberry plants ...

  8. New Method to Disaggregate and Analyze Single Isolated Helminthes Cells Using Flow Cytometry: Proof of Concept

    Directory of Open Access Journals (Sweden)

    Karen Nava-Castro

    2011-01-01

    Full Text Available In parasitology, particularly in helminthes studies, several methods have been used to look for the expression of specific molecules, such as RT-PCR, western blot, 2D-electrophoresis, and microscopy, among others. However, these methods require homogenization of the whole helminth parasite, preventing evaluation of individual cells or specific cell types in a given parasite tissue or organ. Also, the extremely high interaction between helminthes and host cells (particularly immune cells is an important point to be considered. It is really hard to obtain fresh parasites without host cell contamination. Then, it becomes crucial to determine that the analyzed proteins are exclusively from parasitic origin, and not a consequence of host cell contamination. Flow cytometry is a fluorescence-based technique used to evaluate the expression of extra-and intracellular proteins in different type cells, including protozoan parasites. It also allows the isolation and recovery of single-cell populations. Here, we describe a method to isolate and obtain purified helminthes cells.

  9. Inverse PCR-based method for isolating novel SINEs from genome.

    Science.gov (United States)

    Han, Yawei; Chen, Liping; Guan, Lihong; He, Shunping

    2014-04-01

    Short interspersed elements (SINEs) are moderately repetitive DNA sequences in eukaryotic genomes. Although eukaryotic genomes contain numerous SINEs copy, it is very difficult and laborious to isolate and identify them by the reported methods. In this study, the inverse PCR was successfully applied to isolate SINEs from Opsariichthys bidens genome in Eastern Asian Cyprinid. A group of SINEs derived from tRNA(Ala) molecular had been identified, which were named Opsar according to Opsariichthys. SINEs characteristics were exhibited in Opsar, which contained a tRNA(Ala)-derived region at the 5' end, a tRNA-unrelated region, and AT-rich region at the 3' end. The tRNA-derived region of Opsar shared 76 % sequence similarity with tRNA(Ala) gene. This result indicated that Opsar could derive from the inactive or pseudogene of tRNA(Ala). The reliability of method was tested by obtaining C-SINE, Ct-SINE, and M-SINEs from Ctenopharyngodon idellus, Megalobrama amblycephala, and Cyprinus carpio genomes. This method is simpler than the previously reported, which successfully omitted many steps, such as preparation of probes, construction of genomic libraries, and hybridization.

  10. Automatic speech recognition (zero crossing method). Automatic recognition of isolated vowels

    International Nuclear Information System (INIS)

    Dupeyrat, Benoit

    1975-01-01

    This note describes a recognition method of isolated vowels, using a preprocessing of the vocal signal. The processing extracts the extrema of the vocal signal and the interval time separating them (Zero crossing distances of the first derivative of the signal). The recognition of vowels uses normalized histograms of the values of these intervals. The program determines a distance between the histogram of the sound to be recognized and histograms models built during a learning phase. The results processed on real time by a minicomputer, are relatively independent of the speaker, the fundamental frequency being not allowed to vary too much (i.e. speakers of the same sex). (author) [fr

  11. Development of two dimensional electrophoresis method using single chain DNA

    International Nuclear Information System (INIS)

    Ikeda, Junichi; Hidaka, So

    1998-01-01

    By combining a separation method due to molecular weight and a method to distinguish difference of mono-bases, it was aimed to develop a two dimensional single chain DNA labeled with Radioisotope (RI). From electrophoretic pattern difference of parent and variant strands, it was investigated to isolate the root module implantation control gene. At first, a Single Strand Conformation Polymorphism (SSCP) method using concentration gradient gel was investigated. As a result, it was formed that intervals between double chain and single chain DNAs expanded, but intervals of both single chain DNAs did not expand. On next, combination of non-modified acrylic amide electrophoresis method and Denaturing Gradient-Gel Electrophoresis (DGGE) method was examined. As a result, hybrid DNA developed by two dimensional electrophoresis arranged on two lines. But, among them a band of DNA modified by high concentration of urea could not be found. Therefore, in this fiscal year's experiments, no preferable result could be obtained. By the used method, it was thought to be impossible to detect the differences. (G.K.)

  12. Comparative Analysis of the Genomic DNA Isolation Methods on Inula sp. (Asteraceae

    Directory of Open Access Journals (Sweden)

    Emre SEVİNDİK

    2016-12-01

    Full Text Available Simple, fast, low-cost and high throughput protocols are required for DNA isolation of plant species. In this study, phenol chloroform isoamyl alcohol and commercial (Sigma DNA isolation kit methods were applied on some Inula species that belong to Asteraceae family. Genomic DNA amounts, A260, A280, A260/A230 and purity degrees (A260/A280 that were obtained through both methods were measured through electrophoresis and spectrophotometer. Additionally, PCR amplification was realized by primer pairs specific to nrDNA ITS, cpDNA ndhF (972F-1603R and trnL-F regions. Results showed that maximum genomic DNA in nanograms obtained by phenol chloroform isoamyl alcohol method. The study also revealed that I. macrocephala had the maximum DNA and I. heterolepis had the minimum DNA amount. A260/A280 purity degrees showed that the highest and lowest purity in gDNAs obtained through phenol-choloform isoamyl alcohol method were in I.aucheriana and I. salicina, respectively. The highest and lowest purity degrees of gDNAs obtained through commercial kit was observed in I. fragilis and I. macrocephala samples, respectively. PCR amplification results showed that while band profiles of each three regions (ITS, trnL-F and ndhF did not yield positive results in PCR amplifications using phenol-choloform isoamyl alcohol method; PCR band profiles obtained through commercial kit yielded positive results. As a result, it is fair to say that the relation of genomic DNA with PCR was found to be more efficient although the maximum amount of genomic DNA was obtained through phenol chloroform isoamyl alcohol method.

  13. Characterization, thermal stability studies, and analytical method development of Paromomycin for formulation development.

    Science.gov (United States)

    Khan, Wahid; Kumar, Neeraj

    2011-06-01

    Paromomycin (PM) is an aminoglycoside antibiotic, first isolated in the 1950s, and approved in 2006 for treatment of visceral leishmaniasis. Although isolated six decades back, sufficient information essential for development of pharmaceutical formulation is not available for PM. The purpose of this paper was to determine thermal stability and development of new analytical method for formulation development of PM. PM was characterized by thermoanalytical (DSC, TGA, and HSM) and by spectroscopic (FTIR) techniques and these techniques were used to establish thermal stability of PM after heating PM at 100, 110, 120, and 130 °C for 24 h. Biological activity of these heated samples was also determined by microbiological assay. Subsequently, a simple, rapid and sensitive RP-HPLC method for quantitative determination of PM was developed using pre-column derivatization with 9-fluorenylmethyl chloroformate. The developed method was applied to estimate PM quantitatively in two parenteral dosage forms. PM was successfully characterized by various stated techniques. These techniques indicated stability of PM for heating up to 120 °C for 24 h, but when heated at 130 °C, PM is liable to degradation. This degradation is also observed in microbiological assay where PM lost ∼30% of its biological activity when heated at 130 °C for 24 h. New analytical method was developed for PM in the concentration range of 25-200 ng/ml with intra-day and inter-day variability of stability of PM was determined successfully. Developed analytical method was found sensitive, accurate, and precise for quantification of PM. Copyright © 2010 John Wiley & Sons, Ltd. Copyright © 2010 John Wiley & Sons, Ltd.

  14. Developing a TQM quality management method model

    NARCIS (Netherlands)

    Zhang, Zhihai

    1997-01-01

    From an extensive review of total quality management literature, the external and internal environment affecting an organization's quality performance and the eleven primary elements of TQM are identified. Based on the primary TQM elements, a TQM quality management method model is developed. This

  15. Usability Evaluation Method for Agile Software Development

    Directory of Open Access Journals (Sweden)

    Saad Masood Butt

    2015-02-01

    Full Text Available Agile methods are the best fit for tremendously growing software industry due to its flexible and dynamic nature. But the software developed using agile methods do meet the usability standards? To answer this question we can see that majority of agile software development projects currently involve interactive user interface designs, which can only be possible by following User Centered Design (UCD in agile methods. The question here is, how to integrate UCD with agile models. Both Agile models and UCD are iterative in nature but agile models focus on coding and development of software; whereas, UCD focuses on user interface of the software. Similarly, both of them have testing features where the agile model involves automated tested code while UCD involves an expert or a user to test the user interface. In this paper, a new agile usability model is proposed and the evaluation is of the proposed model is presented by practically implementing it in three real life projects. . Key results from these projects clearly show: the proposed agile model incorporates usability evaluation methods, improves the relationship between usability experts to work with agile software experts; in addition, allows agile developers to incorporate the result from UCD into subsequent interactions.

  16. A Framework for Teaching Software Development Methods

    Science.gov (United States)

    Dubinsky, Yael; Hazzan, Orit

    2005-01-01

    This article presents a study that aims at constructing a teaching framework for software development methods in higher education. The research field is a capstone project-based course, offered by the Technion's Department of Computer Science, in which Extreme Programming is introduced. The research paradigm is an Action Research that involves…

  17. Automating Object-Oriented Software Development Methods

    NARCIS (Netherlands)

    Tekinerdogan, B.; Saeki, Motoshi; Sunyé, Gerson; van den Broek, P.M.; Hruby, Pavel; Tekinerdogan, B.; van den Broek, P.M.; Saeki, M.; Hruby, P.; Sunye, G.

    2001-01-01

    Current software projects have generally to deal with producing and managing large and complex software products. It is generally believed that applying software development methods are useful in coping with this complexity and for supporting quality. As such numerous object-oriented software

  18. Automating Object-Oriented Software Development Methods

    NARCIS (Netherlands)

    Tekinerdogan, B.; Frohner, A´ kos; Saeki, Motoshi; Sunyé, Gerson; van den Broek, P.M.; Hruby, Pavel

    2002-01-01

    Current software projects have generally to deal with producing and managing large and complex software products. It is generally believed that applying software development methods are useful in coping with this complexity and for supporting quality. As such numerous object-oriented software

  19. Development of ecofriendly bionanocomposite: Whey protein isolate/pullulan films with nano-SiO2.

    Science.gov (United States)

    Hassannia-Kolaee, Mahbobeh; Khodaiyan, Faramarz; Pourahmad, Rezvan; Shahabi-Ghahfarrokhi, Iman

    2016-05-01

    During the past decade, the limitation of petroleum based polymers, the high price of oil, and the environmental concern were attracted the attention of researchers to develop biobased polymers. The composition of different biopolymers and the reinforcement with nano filler are common methods to improve the drawbacks of biopolymers. In this study whey protein isolate/pullulan (WPI/PUL) films contain 1%, 3%, and 5% (w/w) nano-SiO2 (NS) were prepared by a casting method. Tensile strength of nanocomposite films increased after increasing NS content, but elongation at break decreased, simultaneously. Water absorption, moisture content, solubility in water improved in the wake of increasing NS content because NS increase the cohesiveness of the polymer matrix and improved the barrier and water resistance properties of the films. water vapor permeability of film specimens decreased by increasing NS content. Uniform distribution of NS into polymer matrix was confirmed by scanning electron microscopy (SEM). XRD pattern and thermal analysis revealed increasing crystallinity and increasing Tg of film specimens with increasing NS content, respectively. According to our result WPI/PUL/NS films possess potential to be used as environment friendly packaging films to improve shelf life of food and can be used as promising alternative to petroleum based packaging films. Copyright © 2016 Elsevier B.V. All rights reserved.

  20. Optimal DNA Isolation Method for Detection of Nontuberculous Mycobacteria by Polymerase Chain Reaction

    Directory of Open Access Journals (Sweden)

    Samira Mohammadi

    2017-01-01

    Full Text Available Background: Nontuberculous mycobacteria (NTM are a group of opportunistic pathogens and these are widely dispersed in water and soil resources. Identification of mycobacteria isolates by conventional methods including biochemical tests, growth rates, colony pigmentation, and presence of acid-fast bacilli is widely used, but these methods are time-consuming, labor-intensive, and may sometimes remain inconclusive. Materials and Methods: The DNA was extracted from NTM cultures using CTAB, Chelex, Chelex + Nonidet P-40, FTA® Elute card, and boiling The quantity and quality of the DNA extracted via these methods were determined using UV-photometer at 260 and 280 nm, and polymerase chain reaction (PCR amplification of the heat-shock protein 65 gene with serially diluted DNA samples. Results: The CTAB method showed more positive results at 1:10–1:100,000 at which the DNA amount was substantial. With the Chelex method of DNA extraction, PCR amplification was detected at 1:10 and 1:1000 dilutions. Conclusions: According to the electrophoresis results, the CTAB and Chelex DNA extraction methods were more successful in comparison with the others as regard producing suitable concentrations of DNA with the minimum use of PCR inhibitor.

  1. A Simple and Reproducible Method to Prepare Membrane Samples from Freshly Isolated Rat Brain Microvessels.

    Science.gov (United States)

    Brzica, Hrvoje; Abdullahi, Wazir; Reilly, Bianca G; Ronaldson, Patrick T

    2018-05-07

    The blood-brain barrier (BBB) is a dynamic barrier tissue that responds to various pathophysiological and pharmacological stimuli. Such changes resulting from these stimuli can greatly modulate drug delivery to the brain and, by extension, cause considerable challenges in the treatment of central nervous system (CNS) diseases. Many BBB changes that affect pharmacotherapy, involve proteins that are localized and expressed at the level of endothelial cells. Indeed, such knowledge on BBB physiology in health and disease has sparked considerable interest in the study of these membrane proteins. From a basic science research standpoint, this implies a requirement for a simple but robust and reproducible method for isolation of microvessels from brain tissue harvested from experimental animals. In order to prepare membrane samples from freshly isolated microvessels, it is essential that sample preparations be enriched in endothelial cells but limited in the presence of other cell types of the neurovascular unit (i.e., astrocytes, microglia, neurons, pericytes). An added benefit is the ability to prepare samples from individual animals in order to capture the true variability of protein expression in an experimental population. In this manuscript, details regarding a method that is utilized for isolation of rat brain microvessels and preparation of membrane samples are provided. Microvessel enrichment, from samples derived, is achieved by using four centrifugation steps where dextran is included in the sample buffer. This protocol can easily be adapted by other laboratories for their own specific applications. Samples generated from this protocol have been shown to yield robust experimental data from protein analysis experiments that can greatly aid the understanding of BBB responses to physiological, pathophysiological, and pharmacological stimuli.

  2. PRODUCTION OF EMBBRYONIC STEM CELLS FROM INNER CELL MASS OF BLASTOCYST ISOLATED BY ENZYMATIC AND IMMUNOSURGERY METHODS

    Directory of Open Access Journals (Sweden)

    Thomas Mata Hine

    2008-03-01

    Full Text Available The objective of the research is determining the ICM isolation method to produce ESC. Blastocyst stage of DDy mice embryos were used in this study. Zona pellucida of blastocysts were removed by 0.25% pronase, the ICM isolation were done by enzimatic or immunosurgery method, and then they were cultured in DMEM-high glucose supplemented with mercaptoethanol, gentamycin, fetal bovine serum, and cumulus cells as feeder layer. The result of the research indicated that immunosurgery method yielding attachment rate and number ESC colony 93.85% and 43.08%, respectively, higher (P<0.05 than enzimatic method that weree 79.63% and 18.52%, respectively, but the viability of ICM cells were equal (P >0.05 that are 93.59% in enzymatic method and 98.56% in immunosurgery method. This research concluded that immunosurgery more effective method for isolation of ICM and ESC production than enzymatic method.

  3. Oral rinse as a potential method to culture Candida isolate from AIDS patients

    Directory of Open Access Journals (Sweden)

    Desiana Radithia

    2011-12-01

    Full Text Available Background: Candida isolate is easily sampled from oral cavity by swabbing directly on the candidiasis lesion, to be smeared onto slides for direct examination or cultured in a growth medium. This method is by far the gold standard for defining candidiasis diagnosis. However it is difficult to apply on sensitive patients and almost impossible on patients showing no clinical appearance of oral candidiasis. AIDS patients are very prone to candida infection and have a tendency of repetitive infection involving mixed species. As many candida species show different susceptibility to anti-fungal agents, it is necessary to identify the species causing the infection in the management of oral candidiasis. Oral rinse is a suggested method to obtain candida isolate to be cultured for further analysis such as species identification. This method is simple and less risky on infection transmission as less tools are required in the procedure. Purpose: This study aimed to assess the application of oral rinse as an alternative method to culture Candida isolate from AIDS patients. Methods: A cross-sectional observative study was conducted in HIV/AIDS in-Patient Facility of Intermediate Care Unit for Infection Disease, Dr. Soetomo Hospital Surabaya. Fourteen stadium 4 AIDS patients matching criteria were swabbed on 1/3-posterior of the tongue, and then given 10 ml phosphate buffer saline to rinse vigorously for 15 seconds. Both specimens were cultured on Sabouraud’s dextrose agar and colony growth was observed. Results: Candida colonies were able to grow from all 14 isolates (100% by both methods. Qualitatively, cultures from oral rinse specimens were more populated than cultures from swab specimens. Conclusion: Oral rinse is an applicable technique to obtain Candida species isolate. This technique is safe, easy, non-invasive, and needs less tools therefore less risky for HIV transmission.Latar belakang: Isolat Candida mudah diambil dengan cara mengusap lesi

  4. Development of methods for body composition studies

    International Nuclear Information System (INIS)

    Mattsson, Soeren; Thomas, Brian J

    2006-01-01

    This review is focused on experimental methods for determination of the composition of the human body, its organs and tissues. It summarizes the development and current status of fat determinations from body density, total body water determinations through the dilution technique, whole and partial body potassium measurements for body cell mass estimates, in vivo neutron activation analysis for body protein measurements, dual-energy absorptiometry (DEXA), computed tomography (CT) and magnetic resonance imaging (MRI, fMRI) and spectroscopy (MRS) for body composition studies on tissue and organ levels, as well as single- and multiple-frequency bioimpedance (BIA) and anthropometry as simple easily available methods. Methods for trace element analysis in vivo are also described. Using this wide range of measurement methods, together with gradually improved body composition models, it is now possible to quantify a number of body components and follow their changes in health and disease. (review)

  5. Development of methods for body composition studies

    Energy Technology Data Exchange (ETDEWEB)

    Mattsson, Soeren [Department of Radiation Physics, Lund University, Malmoe University Hospital, SE-205 02 Malmoe (Sweden); Thomas, Brian J [School of Physical and Chemical Sciences, Queensland University of Technology, Brisbane, QLD 4001 (Australia)

    2006-07-07

    This review is focused on experimental methods for determination of the composition of the human body, its organs and tissues. It summarizes the development and current status of fat determinations from body density, total body water determinations through the dilution technique, whole and partial body potassium measurements for body cell mass estimates, in vivo neutron activation analysis for body protein measurements, dual-energy absorptiometry (DEXA), computed tomography (CT) and magnetic resonance imaging (MRI, fMRI) and spectroscopy (MRS) for body composition studies on tissue and organ levels, as well as single- and multiple-frequency bioimpedance (BIA) and anthropometry as simple easily available methods. Methods for trace element analysis in vivo are also described. Using this wide range of measurement methods, together with gradually improved body composition models, it is now possible to quantify a number of body components and follow their changes in health and disease. (review)

  6. Isolation of uv-sensitive variants of human FL cells by a viral suicide method

    International Nuclear Information System (INIS)

    Shiomi, T.; Sato, K.

    1979-01-01

    A new method (viral suicide method) for the isolation of uv-sensitive mutants is described. Colonies of mutagenized human FL cells were infected with uv-irradiated Herpes simplex viruses and surviving ones which seemed to be deficient in host cell reactivation (HCR) were examined for their uv sensitivity. Nineteen of 238 clones examined were sensitive to uv irradiation at the time of the isolation. After recloning, four of these clones have been studied and two (UVS-1 and UVS-2) of them are stable in their uv sensitivity for 4 months in culture. uv sensitivity of UVS-1, UVS-2, and the parental FL cells are as follows: the extrapolation numbers (n) are 2.2, 2.1, and 1.8 and mean lethal doses (DO) are 2.9, 3.7, and 7.8 J/m 2 for UVS-1, UVS-2, and the parental FL cells, respectively. They are no more sensitive than FL cells to x-irradiation. The ability of HCR in UVS-2 cells is apparently lower than that in FL cells, whereas UVS-1 cells are the same as FL cells in the ability

  7. Evaluation of three methods for DNA fingerprinting of Corynebacterium pseudotuberculosis strains isolated from goats in Poland.

    Science.gov (United States)

    Stefańska, Ilona; Rzewuska, Magdalena; Binek, Marian

    2008-01-01

    Phenotypic approaches based on metabolic and biological characteristics of Corynebacterium pseudotuberculosis have been limited due to insufficient discrimination between closely related isolates. In this paper we present performance and convenience of three molecular typing methods: BOX-PCR, random amplification of polymorphic DNA (RAPD) and amplification of DNA fragments surrounding rare restriction site (ADSRRS-fingerprinting) in genome analysis of these bacteria. Among examined 61 strains there were distinguished four, eight and 10 different genotypes by BOX-PCR, RAPD and ADSRRS-fingerprinting, respectively. The value of discrimination index was the lowest for BOX-PCR (D = 0.265), much bigger for RAPD (D = 0.539) and the highest for ADSRRS-fingerprinting (D = 0.604). The good discriminatory ability and reproducibility of RAPD and ADSRRS-fingerprinting indicates that those techniques may be particularly applied for epidemiological studies of C. pseudotuberculosis isolates. We found that ADSRRS-fingerprinting is a rapid method offering good discrimination power, excellent reproducibility and may be applied for epidemiological studies of intraspecific genetic relatedness of C. pseudotuberculosis strains.

  8. Identification of Mucorales isolates from soil using morphological and molecular methods

    Directory of Open Access Journals (Sweden)

    Ardeshir Ziaee

    2016-03-01

    Full Text Available Background and Purpose: Soil is the main habitat of saprophytic and pathogenic fungi. Mucormycetes are one of the most parts of soil fungi and certain members are among opportunistic fungi and can cause systemic fungal infections in immunocompromised patients. The majority of human and animal infections are caused by members of the genera Rhizopus, Mucor, Rhizomucor, Lichtheimia (Absidia, Cunninghamella and Mortierella. The objective of this research was to isolate and identify the main genera of Zygomycetes, using molecular assay and morphological features. Materials and Methods: A total of 340 soil samples were collected from different sites of seven public parks and 14 municipality districts in Isfahan. All samples were cultured on appropriate media and incubated at 27° C for 2 to 4 days, and then examined daily for visible fungal growth. PCR-RFLP method and macroscopic, microscopic and physiological characteristics were applied to identify fungal colonies. Results: Four hundred pure colonies belonging to six genera of Zygomycetes including Lichtheimia, Rhizopus, Rhizomucor, Mucor, Cunninghamella and Mortierella were identified. The genus Rhizopus (35.5% was the most frequent isolate, followed by Mucor (32.25% and Rhizomucor (27.5%. Conclusion: These finding may help us to understand about the importance of opportunistic fungi in public areas and the risk of exposure with immunocompromised persons.

  9. A bootstrapping method for development of Treebank

    Science.gov (United States)

    Zarei, F.; Basirat, A.; Faili, H.; Mirain, M.

    2017-01-01

    Using statistical approaches beside the traditional methods of natural language processing could significantly improve both the quality and performance of several natural language processing (NLP) tasks. The effective usage of these approaches is subject to the availability of the informative, accurate and detailed corpora on which the learners are trained. This article introduces a bootstrapping method for developing annotated corpora based on a complex and rich linguistically motivated elementary structure called supertag. To this end, a hybrid method for supertagging is proposed that combines both of the generative and discriminative methods of supertagging. The method was applied on a subset of Wall Street Journal (WSJ) in order to annotate its sentences with a set of linguistically motivated elementary structures of the English XTAG grammar that is using a lexicalised tree-adjoining grammar formalism. The empirical results confirm that the bootstrapping method provides a satisfactory way for annotating the English sentences with the mentioned structures. The experiments show that the method could automatically annotate about 20% of WSJ with the accuracy of F-measure about 80% of which is particularly 12% higher than the F-measure of the XTAG Treebank automatically generated from the approach proposed by Basirat and Faili [(2013). Bridge the gap between statistical and hand-crafted grammars. Computer Speech and Language, 27, 1085-1104].

  10. Optimal DNA Isolation Method for Detection of Nontuberculous Mycobacteria by Polymerase Chain Reaction.

    Science.gov (United States)

    Mohammadi, Samira; Esfahani, Bahram Nasr; Moghim, Sharareh; Mirhendi, Hossein; Zaniani, Fatemeh Riyahi; Safaei, Hajieh Ghasemian; Fazeli, Hossein; Salehi, Mahshid

    2017-01-01

    Nontuberculous mycobacteria (NTM) are a group of opportunistic pathogens and these are widely dispersed in water and soil resources. Identification of mycobacteria isolates by conventional methods including biochemical tests, growth rates, colony pigmentation, and presence of acid-fast bacilli is widely used, but these methods are time-consuming, labor-intensive, and may sometimes remain inconclusive. The DNA was extracted from NTM cultures using CTAB, Chelex, Chelex + Nonidet P-40, FTA ® Elute card, and boiling The quantity and quality of the DNA extracted via these methods were determined using UV-photometer at 260 and 280 nm, and polymerase chain reaction (PCR) amplification of the heat-shock protein 65 gene with serially diluted DNA samples. The CTAB method showed more positive results at 1:10-1:100,000 at which the DNA amount was substantial. With the Chelex method of DNA extraction, PCR amplification was detected at 1:10 and 1:1000 dilutions. According to the electrophoresis results, the CTAB and Chelex DNA extraction methods were more successful in comparison with the others as regard producing suitable concentrations of DNA with the minimum use of PCR inhibitor.

  11. Stochastic development regression using method of moments

    DEFF Research Database (Denmark)

    Kühnel, Line; Sommer, Stefan Horst

    2017-01-01

    This paper considers the estimation problem arising when inferring parameters in the stochastic development regression model for manifold valued non-linear data. Stochastic development regression captures the relation between manifold-valued response and Euclidean covariate variables using...... the stochastic development construction. It is thereby able to incorporate several covariate variables and random effects. The model is intrinsically defined using the connection of the manifold, and the use of stochastic development avoids linearizing the geometry. We propose to infer parameters using...... the Method of Moments procedure that matches known constraints on moments of the observations conditional on the latent variables. The performance of the model is investigated in a simulation example using data on finite dimensional landmark manifolds....

  12. Improved method for rapid and accurate isolation and identification of Streptococcus mutans and Streptococcus sobrinus from human plaque samples.

    Science.gov (United States)

    Villhauer, Alissa L; Lynch, David J; Drake, David R

    2017-08-01

    Mutans streptococci (MS), specifically Streptococcus mutans (SM) and Streptococcus sobrinus (SS), are bacterial species frequently targeted for investigation due to their role in the etiology of dental caries. Differentiation of S. mutans and S. sobrinus is an essential part of exploring the role of these organisms in disease progression and the impact of the presence of either/both on a subject's caries experience. Of vital importance to the study of these organisms is an identification protocol that allows us to distinguish between the two species in an easy, accurate, and timely manner. While conducting a 5-year birth cohort study in a Northern Plains American Indian tribe, the need for a more rapid procedure for isolating and identifying high volumes of MS was recognized. We report here on the development of an accurate and rapid method for MS identification. Accuracy, ease of use, and material and time requirements for morphological differentiation on selective agar, biochemical tests, and various combinations of PCR primers were compared. The final protocol included preliminary identification based on colony morphology followed by PCR confirmation of species identification using primers targeting regions of the glucosyltransferase (gtf) genes of SM and SS. This method of isolation and identification was found to be highly accurate, more rapid than the previous methodology used, and easily learned. It resulted in more efficient use of both time and material resources. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. A method of experimental rheumatoid arthritis induction using collagen type II isolated from chicken sternal cartilage.

    Science.gov (United States)

    Su, Zhaoliang; Shotorbani, Siamak Sandoghchian; Jiang, Xugan; Ma, Rui; Shen, Huiling; Kong, Fanzhi; Xu, Huaxi

    2013-07-01

    At present, collagen‑induced arthritis (CIA) is the best known and most extensively used model for the immunological and pathological characteristics of human rheumatoid arthritis (RA). This model is useful not only in aiding our understanding of the pathogenesis of this disease, but also in the development of new therapies. Bovine, porcine and human collagen has been used to induce CIA; however, response has been identified to vary between strains and injection conditions, and false positive results and reduced potency are common as a result of minor contaminants or deglycosylated protein. Therefore, in the present study, type II collagen (CII) was isolated and purified from chicken sternal cartilage and was found to successfully induce the RA model. Furthermore, T helper 17 (Th17) cells were observed to infiltrate the joint on day 45 following induction by CII. In vitro, expression of toll‑like receptor 2 (TLR2) increased in peritoneal macrophages stimulated by CII. In addition, blockage of TLR2 was identified to markedly decrease levels of TGF‑β and IL‑6 in the cell culture supernatant. The results indicate that CII isolated from chicken sternal cartilage may be recognized by TLR2 on macrophages, leading to TGF‑β and IL‑6 production and subsequent activation of Th17 cells which mediates CIA development.

  14. Scientific methods for developing ultrastable structures

    International Nuclear Information System (INIS)

    Gamble, M.; Thompson, T.; Miller, W.

    1990-01-01

    Scientific methods used by the Los Alamos National Laboratory for developing an ultrastable structure for study of silicon-based elementary particle tracking systems are addressed. In particular, the design, analysis, and monitoring of this system are explored. The development methodology was based on a triad of analytical, computational, and experimental techniques. These were used to achieve a significant degree of mechanical stability (alignment accuracy >1 μrad) and yet allow dynamic manipulation of the system. Estimates of system thermal and vibratory stability and component performance are compared with experimental data collected using laser interferometry and accelerometers. 8 refs., 5 figs., 4 tabs

  15. Thrombus Development on a Transseptal Sheath in the Right Atrium Before Electrical Pulmonary Vein Isolation

    Directory of Open Access Journals (Sweden)

    Nieves Romero-Rodriguez

    2008-02-01

    Full Text Available We describe the case of a patient who developed a thrombus on the transseptal sheath in the right atrium before transseptal puncture for circumferential pulmonary vein isolation for paroxysmal atrial fibrillation treatment. The use of intracardiac echocardiography allowed to its identification and probably prevented the patient from suffering a serious thromboembolic complication.

  16. Job-Embedded Professional Development: Reducing Teacher Isolation by Enacting Social Change

    Science.gov (United States)

    Frank, K.

    2009-01-01

    Teacher isolation and burn-out are problems which contribute to high rates of teacher attrition and require school districts to hire new teachers each year and start anew with professional development. The purpose of this qualitative case study was to investigate whether peer coaching and mentoring programs had an effect on this problem by…

  17. Seismic isolation development for the US advanced liquid-metal reactor program

    International Nuclear Information System (INIS)

    Gluekler, E.L.; Bigelow, C.C.; DeVita, V.; Kelly, J.M.; Seidensticker, R.W.; Tajirian, F.F.

    1989-01-01

    GE Nuclear Energy, in association with a US Industrial Team and support from the US National Laboratories and Universities, is developing a modular liquid-metal reactor concept for the US Department of Energy (DOE). The objective of this development is to provide, by the turn of the century, a reactor concept with optimized passive safety features that is economically competitive with other domestic energy sources, licensable, and ready for commercial deployment. One of the unique features of the concept is the seismic isolation of the reactor modules which decouples the reactor and their safety systems from potentially damaging ground motions and significantly enhances the structural resistance to high energy, as well as long duration earthquakes. Seismic isolation is accomplished with high damping natural rubber bearings. The reactors are located in individual silos below grade level and are supported by the isolator bearings at approximately their center of gravity. This application of seismic isolation is the first for a US nuclear power plant. A development program has been established to assure the full benefits from the utilization of this new approach and to provide adequate system characterization and qualification for licensing certification. The development program is described in this paper and selected results are presented. The initial testing indicated excellent performance of high damping natural rubber bearings

  18. Development of a transient criticality evaluation method

    International Nuclear Information System (INIS)

    Pain, C.C.; Eaton, M.D.; Miles, B.; Ziver, A.K.; Gomes, J.L.M.A.; Umpleby, A.P.; Piggott, M.D.; Goddard, A.J.H.; Oliveira, C.R.E. de

    2005-01-01

    In developing a transient criticality evaluation method we model, in full spatial/temporal detail, the neutron fluxes and consequent power and the evolving material properties - their flows, energies, phase changes etc. These methods are embodied in the generic method FETCH code which is based as far as possible on basic principles and is capable of use in exploring safety-related situations somewhat beyond the range of experiment. FETCH is a general geometry code capable of addressing a range of criticality issues in fissile materials. The code embodies both transient radiation transport and transient fluid dynamics. Work on powders, granular materials, porous media and solutions is reviewed. The capability for modelling transient criticality for chemical plant, waste matrices and advanced reactors is also outlined. (author)

  19. Summary of the systems prioritization method as a decision-aiding method for the waste isolation pilot plant

    International Nuclear Information System (INIS)

    Boak, D.M.; Prindle, N.H.; Lincoln, R.

    1996-12-01

    In March 1994, the U.S. Department of Energy Carlsbad Area Office (DOE/CAO) implemented a performance-based decision-aiding method to assist in programmatic prioritization within the Waste Isolation Pilot Plant (WIPP) Project with respect to applicable U.S. Environmental Protection Agency (EPA) long-term performance requirements in 40 CFR 191.13(a) (radionuclide containment requirements) and 40 CFR 268.6 (hazardous constituent concentration requirements). This method, the Systems Prioritization Method (SPM), was designed by Sandia National Laboratories (SNL) to: (1) identify programmatic options (activities) and their costs and durations; (2) analyze combinations of activities (activity sets) in terms of their predicted contribution to long-term performance of the WIPP disposal system; and (3) analyze cost, duration, and performance tradeoffs. The results of the second iteration of SPM (SPM-2) were the basis for recommendations to DOE/CAO in May 1995 for programmatic prioritization within the WIPP project. This paper presents a summary of the SPM implementation, key results, and lessons learned

  20. Summary of the systems prioritization method as a decision-aiding method for the waste isolation pilot plant

    Energy Technology Data Exchange (ETDEWEB)

    Boak, D.M.; Prindle, N.H.; Lincoln, R. [and others

    1996-12-01

    In March 1994, the U.S. Department of Energy Carlsbad Area Office (DOE/CAO) implemented a performance-based decision-aiding method to assist in programmatic prioritization within the Waste Isolation Pilot Plant (WIPP) Project with respect to applicable U.S. Environmental Protection Agency (EPA) long-term performance requirements in 40 CFR 191.13(a) (radionuclide containment requirements) and 40 CFR 268.6 (hazardous constituent concentration requirements). This method, the Systems Prioritization Method (SPM), was designed by Sandia National Laboratories (SNL) to: (1) identify programmatic options (activities) and their costs and durations; (2) analyze combinations of activities (activity sets) in terms of their predicted contribution to long-term performance of the WIPP disposal system; and (3) analyze cost, duration, and performance tradeoffs. The results of the second iteration of SPM (SPM-2) were the basis for recommendations to DOE/CAO in May 1995 for programmatic prioritization within the WIPP project. This paper presents a summary of the SPM implementation, key results, and lessons learned.

  1. Seismic isolation development for the US advanced liquid-metal reactor program

    International Nuclear Information System (INIS)

    Gluekler, E.L.; Bigelow, C.C.; DeVita, V.; Kelly, J.M.; Seidensticker, R.W.; Tajirian, F.F.

    1991-01-01

    GE Nuclear Energy, in association with a US Industrial Team and support from the US National Laboratories and Universities, is developing a modular liquid-metal reactor concept for the US DOE. The objective of this development is to provide, by the turn of the century, a reactor with optimized passive safety features that is economically competitive with other domestic energy sources, licensable, and ready for commercial deployment. One of the unique features of the concept is the seismic isolation of the reactor modules which decouples the reactors and their safety systems from potentially damaging ground motions and significantly enhances the structural resistance to high energy, as well as long-duration earthquakes. Seismic isolation is accomplished with high-damping natural-rubber bearings. The reactors are located in individual silos below grade level and are supported by the isolator bearings at approximately their center of gravity. This application of seismic isolation is the first for a US nuclear power plant. A development program has been established to assure the full benefits from the utilization of this new approach and to provide adequate system characterization and qualification for licensing certification. The development program, which is supported by the US DOE, ANL, Energy Technology Engineering Center (ETEC), the University of California at Berkeley (UC-Berkeley), GE, and Bechtel National, Inc. (BNI), is described and selected results are presented. The initial testing indicated excellent performance of high-damping natural-rubber bearings. The development of seismic isolation guidelines is in progress as a joint activity between ENEA of Italy and the GE Team. (orig./HP)

  2. Standardization of sample collection, isolation and analysis methods in extracellular vesicle research

    Directory of Open Access Journals (Sweden)

    Kenneth W. Witwer

    2013-05-01

    Full Text Available The emergence of publications on extracellular RNA (exRNA and extracellular vesicles (EV has highlighted the potential of these molecules and vehicles as biomarkers of disease and therapeutic targets. These findings have created a paradigm shift, most prominently in the field of oncology, prompting expanded interest in the field and dedication of funds for EV research. At the same time, understanding of EV subtypes, biogenesis, cargo and mechanisms of shuttling remains incomplete. The techniques that can be harnessed to address the many gaps in our current knowledge were the subject of a special workshop of the International Society for Extracellular Vesicles (ISEV in New York City in October 2012. As part of the “ISEV Research Seminar: Analysis and Function of RNA in Extracellular Vesicles (evRNA”, 6 round-table discussions were held to provide an evidence-based framework for isolation and analysis of EV, purification and analysis of associated RNA molecules, and molecular engineering of EV for therapeutic intervention. This article arises from the discussion of EV isolation and analysis at that meeting. The conclusions of the round table are supplemented with a review of published materials and our experience. Controversies and outstanding questions are identified that may inform future research and funding priorities. While we emphasize the need for standardization of specimen handling, appropriate normative controls, and isolation and analysis techniques to facilitate comparison of results, we also recognize that continual development and evaluation of techniques will be necessary as new knowledge is amassed. On many points, consensus has not yet been achieved and must be built through the reporting of well-controlled experiments.

  3. Isolation and purification of Echinococcus granulosus antigen B from hydatid cyst fluid using three different methods

    Directory of Open Access Journals (Sweden)

    Shirazi, S.

    2016-07-01

    Full Text Available Hydatid cyst, the larval stage of cestodes Echinococcus spp., is recognized as a zoonotic infection in the world. The World Health Organization (WHO has recently classified echinococcosis in a group of neglected tropical diseases. The prevalence of Echinococcus granulosus infection is high in Iran due to the presence of various intermediate hosts in this country. Considering the rising trend of this zoonotic parasitic disease based on national epidemiological studies, diagnosis is of great significance. WHO has suggested the use of specific antigens, especially antigen B (AgB for serological diagnostic tests. In general, AgB is a polymeric lipoprotein, which disintegrates into 8.12, 16, and 20.24 kDa subunits. In the present study, we applied three different methods for AgB isolation from hydatid cyst fluid (HCF and compared their efficacy in AgB isolation. Finally, the protein concentration of this antigen was measured by Bradford assay and confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE. The results showed that the application of polyethylene glycol (PEG 4000 as a thickener agent beside purification of HCF in dialysis bag and filtering and also dialysis against acetate buffer leading to the best quantity in purified antigen B.

  4. Identification of Mucorales isolates from soil using morphological and molecular methods.

    Science.gov (United States)

    Ziaee, A; Zia, M; Bayat, M; Hashemi, J

    2016-03-01

    Soil is the main habitat of saprophytic and pathogenic fungi. Mucoromycotina constitutes a large group of soil fungi, with certain opportunistic members causing systemic infections in immunocompromised hosts. The majority of human and animal infections are caused by the members of the genera Rhizopus , Mucor , Rhizomucor , Lichtheimia ( Absidia) , Cunninghamella, and Mortierella . Accordingly, in the present study, we aimed to isolate and identify the main genera of the order Mucorales, using molecular assays and morphological features . In total, 340 soil samples were collected from seven public parks throughout the city and sidewalk gardens in 14 municipal districts in Isfahan, Iran. All the samples were cultured on the appropriate media, incubated at 27°C for 2- 4 days, and examined daily for visible fungal growth. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was applied and macroscopic, microscopic, and physiological characteristics were assessed to identify fungal colonies. 400 pure colonies, belonging to the orders Mucorales and Mortierellales, including the genera Lichtheimia , Rhizopus, Rhizomucor, Mucor, Cunninghamella, and Mortierella, were identified . The genus Rhizopus (35.5%) was the most frequent isolate, followed by Mucor (32.25%) and Rhizomucor (27.5%). The results emphasize the importance of opportunistic fungi in public areas and indicate the risk of exposure for immunocompromised individuals.

  5. Isolation, characterization and development of bacteria in the Mine Gafsa for applications in bioremediation

    International Nuclear Information System (INIS)

    Heni, Sana

    2010-01-01

    Today pollution represents an important environmental problem. Bacterial ability to bioremediate many types of pollutants in different matrixes (soil, water, and air) have been widely acknowledged. The goal of the present work is to isolate from contaminated soil of Gafsa, in Tunisia, bacterial strains to evaluate their potential for bioremediation. Soil from the mining area of Gafsa was collected. Initially, many bacterial strains were isolated in TGY agar (Tryptone/Glucose/Yeast extract agar) based on the presence of pigments. The primary bacterial selection was performed using heavy metals and the minimal inhibitory concentrations (MICs) of a metal-resistant bacterium, Cupriavidus metallidurans CH34. Isolated metal-resistant bacterium was checked for its potential to resistant to gamma radiation. Selected strain, Micrococcus luteus S7, was assessed for its bioremediation potential of matrixes artificially contaminated under laboratory conditions for its future use in developing a bio product for contaminated soil inoculation.

  6. Numerical Solutions for Nonlinear High Damping Rubber Bearing Isolators: Newmark's Method with Netwon-Raphson Iteration Revisited

    Science.gov (United States)

    Markou, A. A.; Manolis, G. D.

    2018-03-01

    Numerical methods for the solution of dynamical problems in engineering go back to 1950. The most famous and widely-used time stepping algorithm was developed by Newmark in 1959. In the present study, for the first time, the Newmark algorithm is developed for the case of the trilinear hysteretic model, a model that was used to describe the shear behaviour of high damping rubber bearings. This model is calibrated against free-vibration field tests implemented on a hybrid base isolated building, namely the Solarino project in Italy, as well as against laboratory experiments. A single-degree-of-freedom system is used to describe the behaviour of a low-rise building isolated with a hybrid system comprising high damping rubber bearings and low friction sliding bearings. The behaviour of the high damping rubber bearings is simulated by the trilinear hysteretic model, while the description of the behaviour of the low friction sliding bearings is modeled by a linear Coulomb friction model. In order to prove the effectiveness of the numerical method we compare the analytically solved trilinear hysteretic model calibrated from free-vibration field tests (Solarino project) against the same model solved with the Newmark method with Netwon-Raphson iteration. Almost perfect agreement is observed between the semi-analytical solution and the fully numerical solution with Newmark's time integration algorithm. This will allow for extension of the trilinear mechanical models to bidirectional horizontal motion, to time-varying vertical loads, to multi-degree-of-freedom-systems, as well to generalized models connected in parallel, where only numerical solutions are possible.

  7. Numerical Solutions for Nonlinear High Damping Rubber Bearing Isolators: Newmark’s Method with Netwon-Raphson Iteration Revisited

    Directory of Open Access Journals (Sweden)

    Markou A.A.

    2018-03-01

    Full Text Available Numerical methods for the solution of dynamical problems in engineering go back to 1950. The most famous and widely-used time stepping algorithm was developed by Newmark in 1959. In the present study, for the first time, the Newmark algorithm is developed for the case of the trilinear hysteretic model, a model that was used to describe the shear behaviour of high damping rubber bearings. This model is calibrated against free-vibration field tests implemented on a hybrid base isolated building, namely the Solarino project in Italy, as well as against laboratory experiments. A single-degree-of-freedom system is used to describe the behaviour of a low-rise building isolated with a hybrid system comprising high damping rubber bearings and low friction sliding bearings. The behaviour of the high damping rubber bearings is simulated by the trilinear hysteretic model, while the description of the behaviour of the low friction sliding bearings is modeled by a linear Coulomb friction model. In order to prove the effectiveness of the numerical method we compare the analytically solved trilinear hysteretic model calibrated from free-vibration field tests (Solarino project against the same model solved with the Newmark method with Netwon-Raphson iteration. Almost perfect agreement is observed between the semi-analytical solution and the fully numerical solution with Newmark’s time integration algorithm. This will allow for extension of the trilinear mechanical models to bidirectional horizontal motion, to time-varying vertical loads, to multi-degree-of-freedom-systems, as well to generalized models connected in parallel, where only numerical solutions are possible.

  8. Isolating Trait and Method Variance in the Measurement of Callous and Unemotional Traits.

    Science.gov (United States)

    Paiva-Salisbury, Melissa L; Gill, Andrew D; Stickle, Timothy R

    2017-09-01

    To examine hypothesized influence of method variance from negatively keyed items in measurement of callous-unemotional (CU) traits, nine a priori confirmatory factor analysis model comparisons of the Inventory of Callous-Unemotional Traits were evaluated on multiple fit indices and theoretical coherence. Tested models included a unidimensional model, a three-factor model, a three-bifactor model, an item response theory-shortened model, two item-parceled models, and three correlated trait-correlated method minus one models (unidimensional, correlated three-factor, and bifactor). Data were self-reports of 234 adolescents (191 juvenile offenders, 43 high school students; 63% male; ages 11-17 years). Consistent with hypotheses, models accounting for method variance substantially improved fit to the data. Additionally, bifactor models with a general CU factor better fit the data compared with correlated factor models, suggesting a general CU factor is important to understanding the construct of CU traits. Future Inventory of Callous-Unemotional Traits analyses should account for method variance from item keying and response bias to isolate trait variance.

  9. Isolation of full-length putative rat lysophospholipase cDNA using improved methods for mRNA isolation and cDNA cloning

    International Nuclear Information System (INIS)

    Han, J.H.; Stratowa, C.; Rutter, W.J.

    1987-01-01

    The authors have cloned a full-length putative rat pancreatic lysophospholipase cDNA by an improved mRNA isolation method and cDNA cloning strategy using [ 32 P]-labelled nucleotides. These new methods allow the construction of a cDNA library from the adult rat pancreas in which the majority of recombinant clones contained complete sequences for the corresponding mRNAs. A previously recognized but unidentified long and relatively rare cDNA clone containing the entire sequence from the cap site at the 5' end to the poly(A) tail at the 3' end of the mRNA was isolated by single-step screening of the library. The size, amino acid composition, and the activity of the protein expressed in heterologous cells strongly suggest this mRNA codes for lysophospholipase

  10. Prioritizing pesticide compounds for analytical methods development

    Science.gov (United States)

    Norman, Julia E.; Kuivila, Kathryn; Nowell, Lisa H.

    2012-01-01

    The U.S. Geological Survey (USGS) has a periodic need to re-evaluate pesticide compounds in terms of priorities for inclusion in monitoring and studies and, thus, must also assess the current analytical capabilities for pesticide detection. To meet this need, a strategy has been developed to prioritize pesticides and degradates for analytical methods development. Screening procedures were developed to separately prioritize pesticide compounds in water and sediment. The procedures evaluate pesticide compounds in existing USGS analytical methods for water and sediment and compounds for which recent agricultural-use information was available. Measured occurrence (detection frequency and concentrations) in water and sediment, predicted concentrations in water and predicted likelihood of occurrence in sediment, potential toxicity to aquatic life or humans, and priorities of other agencies or organizations, regulatory or otherwise, were considered. Several existing strategies for prioritizing chemicals for various purposes were reviewed, including those that identify and prioritize persistent, bioaccumulative, and toxic compounds, and those that determine candidates for future regulation of drinking-water contaminants. The systematic procedures developed and used in this study rely on concepts common to many previously established strategies. The evaluation of pesticide compounds resulted in the classification of compounds into three groups: Tier 1 for high priority compounds, Tier 2 for moderate priority compounds, and Tier 3 for low priority compounds. For water, a total of 247 pesticide compounds were classified as Tier 1 and, thus, are high priority for inclusion in analytical methods for monitoring and studies. Of these, about three-quarters are included in some USGS analytical method; however, many of these compounds are included on research methods that are expensive and for which there are few data on environmental samples. The remaining quarter of Tier 1

  11. Research of isolated resonances using the average energy shift method for filtered neutron beam

    International Nuclear Information System (INIS)

    Gritzay, O.O.; Grymalo, A.K.; Kolotyi, V.V.; Mityushkin, O.O.; Venediktov, V.M.

    2010-01-01

    This work is devoted to detailed description of one of the research directions in the Neutron Physics Department (NPD), namely, to research of resonance parameters of isolated nuclear level at the filtered neutron beam on the horizontal experimental channel HEC-8 of the WWR-M reactor. Research of resonance parameters is an actual problem nowadays. This is because there are the essential differences between the resonance parameter values in the different evaluated nuclear data library (ENDL) for many nuclei. Research of resonance parameter is possible due to the set of the neutron cross sections received at the same filter, but with the slightly shifted filter average energy. The shift of the filter average energy is possible by several processes. In this work this shift is realized by neutron energy dependence on scattering angle. This method is provided by equipment.

  12. A rapid and simple method for DNA extraction from yeasts and fungi isolated from Agave fourcroydes.

    Science.gov (United States)

    Tapia-Tussell, Raul; Lappe, Patricia; Ulloa, Miguel; Quijano-Ramayo, Andrés; Cáceres-Farfán, Mirbella; Larqué-Saavedra, Alfonso; Perez-Brito, Daisy

    2006-05-01

    A simple and easy protocol for extracting high-quality DNA from different yeast and filamentous fungal species is described. This method involves two important steps: first, the disruption of cell walls by mechanical means and freezing; and second, the extraction, isolation, and precipitation of genomic DNA. The absorbance ratios (A(260)/A(280)) obtained ranged from 1.6 to 2.0. The main objective of this procedure is to extract pure DNA from yeast and filamentous fungi, including those with high contents of proteins, polysaccharides, and other complex compounds in their cell walls. The yield and quality of the DNAs obtained were suitable for micro/minisatellite primer-polymerase chain reaction (MSP-PCR) fingerprinting as well as for the sequence of the D1/D2 domain of the 26S rDNA.

  13. Properties of cellulose nanocrystals from oil palm trunk isolated by total chlorine free method.

    Science.gov (United States)

    Lamaming, Junidah; Hashim, Rokiah; Leh, Cheu Peng; Sulaiman, Othman

    2017-01-20

    Cellulose nanocrystals were isolated from oil palm trunk by total chlorine free method. The samples were either water pre-hydrolyzed or non-water pre-hydrolyzed, subjected to soda pulping, acidified and ozone bleached. Cellulose and cellulose nanocrystal (CNC) physical, chemical, thermal properties, and crystallinity index were investigated by composition analysis, scanning electron microscopy, transmission electron microscopy, fourier transform infrared, thermogravimetric analysis and X-ray diffraction. Water pre-hydrolysis reduced lignin (process compared to non-fibrillated of non-water pre-hydrolyzed cellulose. Water pre-hydrolysis improved final CNC crystallinity (up to 75%) compared to CNC without water pre-hydrolysis crystallinity (69%). Cellulose degradation was found to occur during ozone bleaching stage but CNC showed an increase in crystallinity after acid hydrolysis. Thus, oil palm trunk CNC can be potentially applied in pharmaceutical, food, medical and nanocomposites. Copyright © 2016 Elsevier Ltd. All rights reserved.

  14. A well-posed numerical method to track isolated conformal map singularities in Hele-Shaw flow

    International Nuclear Information System (INIS)

    Baker, G.; Siegel, M.; Tanveer, S.

    1995-01-01

    We present a new numerical method for calculating an evolving 2D Hele-Shaw interface when surface tension effects are neglected. In the case where the flow is directed from the less viscous fluid into the more viscous fluid, the motion of the interface is ill-posed; small deviations in the initial condition will produce significant changes in the ensuing motion. The situation is disastrous for numerical computation, as small roundoff errors can quickly lead to large inaccuracies in the computed solution. Our method of computation is most easily formulated using a conformal map from the fluid domain into a unit disk. The method relies on analytically continuing the initial data and equations of motion into the region exterior to the disk, where the evolution problem becomes well-posed. The equations are then numerically solved in the extended domain. The presence of singularities in the conformal map outside of the disk introduces specific structures along the fluid interface. Our method can explicitly track the location of isolated pole and branch point singularities, allowing us to draw connections between the development of interfacial patterns and the motion of singularities as they approach the unit disk. In particular, we are able to relate physical features such as finger shape, side-branch formation, and competition between fingers to the nature and location of the singularities. The usefulness of this method in studying the formation of topological singularities (self-intersections of the interface) is also pointed out. 47 refs., 10 figs., 1 tab

  15. Shielding methods development in the United States

    International Nuclear Information System (INIS)

    Mynatt, F.R.

    1977-01-01

    A generalized shielding methodology has been developed in the U.S.A. that is adaptable to the shielding analyses of all reactor types. Thus far used primarily for liquid-metal fast breeder reactors, the methodology includes several component activities: (1) developing methods for calculating radiation transport through reactor-shield systems; (2) processing cross-section libraries; (3) performing design calculations for specific systems; (4) performing and analyzing pertinent integral experiments; (5) performing sensitivity studies on both the design calculations and the experimental analyses; and, finally, (6) calculating shield design parameters and their uncertainties. The criteria for the methodology are a 5 to 10 percent accuracy for responses at locations near the core and a factor of 2 accuracy for responses at distant locations. The methodology has been successfully adapted to most in-vessel and ex-vessel problems encountered in the shield analyses of the Fast Flux Test Facility and the Fast Flux Test Facility and the Clinch River Breeder Reactor; however, improved techniques are needed for calculating regions in which radiation streaming is dominant. Areas of the methodology in which significant progress has recently been made are those involving the development of cross-section libraries, sensitivity analysis methods, and transport codes

  16. New method of isolation and perfusion of rat pancreas for phosphorus nucletic resonance study

    Energy Technology Data Exchange (ETDEWEB)

    Matsumoto, Takehisa; Kanno, Tomio; Murakami, Masataka; Watari, Hiroshi; Seo, Yoshiteru

    1988-02-01

    Phosphorus nuclear magnetic resonance (/sup 31/P-NMR) was applied to measure phosphorus metabolites in the pancreas. Rat pancreatic gland was isolated and perfused with a modified Krebs-Henseleit solution at room temperature (25 deg C). Removal of the duodenum from the pancreas made it possible to record /sup 31/P-NMR spectra only from the pancreatic tissue. /sup 31/P signals of creatine phosphate (PCr), ATP and inorganic phosphate (Pi) were observed in all the glands. In different individual glands, the Signal intensity of PCr varied, but those of ATP were almost the same. The initial levels of all the phosphorus metabolites in individual glands remained almost unchanged during perfusion with the standard solution for 2 hours. When the perfusion was stopped, signal intensity of ATP and PCr decreased while Pi resonance increased and shifted to a higher magnetic field, indicating tissue acidification. On re-perfusion, each resonance and tissue pH were restored to the respective initial resting levels. Continuous stimulation with 0.3 ..mu..M acetylcholine (ACh) or 0.1 nM CCK-8 caused a markedly larger increase in pancreatic exocrine secretion than that observed in the perfused pancreas which was prepared by the previous method. Continuous stimulation with 0.1 ..mu..M ACh caused a marked and sustained increase in pancreatic exocrine secretion. During the stimulation, the resonances of phosphorus energy metabolites and tissue pH remained almost unchanged. These results indicate that the isolated perfused rat pancreas which is prepared by our new method is a useful preparation for analysis of secretion-metabolism coupling in the exocrine pancreas.

  17. Structural reliability methods: Code development status

    Science.gov (United States)

    Millwater, Harry R.; Thacker, Ben H.; Wu, Y.-T.; Cruse, T. A.

    1991-05-01

    The Probabilistic Structures Analysis Method (PSAM) program integrates state of the art probabilistic algorithms with structural analysis methods in order to quantify the behavior of Space Shuttle Main Engine structures subject to uncertain loadings, boundary conditions, material parameters, and geometric conditions. An advanced, efficient probabilistic structural analysis software program, NESSUS (Numerical Evaluation of Stochastic Structures Under Stress) was developed as a deliverable. NESSUS contains a number of integrated software components to perform probabilistic analysis of complex structures. A nonlinear finite element module NESSUS/FEM is used to model the structure and obtain structural sensitivities. Some of the capabilities of NESSUS/FEM are shown. A Fast Probability Integration module NESSUS/FPI estimates the probability given the structural sensitivities. A driver module, PFEM, couples the FEM and FPI. NESSUS, version 5.0, addresses component reliability, resistance, and risk.

  18. Developments of an Interactive Sail Design Method

    OpenAIRE

    S. M. Malpede; M. Vezza

    2000-01-01

    This paper presents a new tool for performing the integrated design and analysis of a sail. The features of the system are the geometrical definition of a sail shape, using the Bezier surface method, the creation of a finite element model for the non-linear structural analysis and a fluid-dynamic model for the aerodynamic analysis. The system has been developed using MATLAB(r). Recent sail design efforts have been focused on solving the aeroelastic behavior of the sail. The pressure dis...

  19. Sea sand disruption method (SSDM) as a valuable tool for isolating essential oil components from conifers.

    Science.gov (United States)

    Dawidowicz, Andrzej L; Czapczyńska, Natalia B

    2011-11-01

    Essential oils are one of nature's most precious gifts with surprisingly potent and outstanding properties. Coniferous oils, for instance, are nowadays being used extensively to treat or prevent many types of infections, modify immune responses, soothe inflammations, stabilize moods, and to help ease all forms of non-acute pain. Given the broad spectrum of usage of coniferous essential oils, a fast, safe, simple, and efficient sample-preparation method is needed in the estimation procedure of essential oil components in fresh plant material. Generally, the time- and energy-consuming steam distillation (SD) is applied for this purpose. This paper will compare SD, pressurized liquid extraction (PLE), matrix solid-phase dispersion (MSPD), and the sea sand disruption method (SSDM) as isolation techniques to obtain aroma components from Scots pine (Pinus sylvestris), spruce (Picea abies), and Douglas fir (Pseudotsuga menziesii). According to the obtained data, SSDM is the most efficient sample preparation method in determining the essential oil composition of conifers. Moreover, SSDM requires small organic solvent amounts and a short extraction time, which makes it an advantageous alternative procedure for the routine analysis of coniferous oils. The superiority of SSDM over MSPD efficiency is ascertained, as there are no chemical interactions between the plant cell components and the sand. This fact confirms the reliability and efficacy of SSDM for the analysis of volatile oil components. Copyright © 2011 Verlag Helvetica Chimica Acta AG, Zürich.

  20. The Basalt Waste Isolation Project technical program evaluation process: A criteria-based method

    International Nuclear Information System (INIS)

    Babad, H.; Evans, C.; Wolfe, B.A.

    1982-01-01

    The need to objectively evaluate the progress being made by the Basalt Waste Isolation Project (BWIP) toward establishing the feasibility of siting a nuclear waste repository in basalt (NWRB) mandates a process for evaluating the technical work of the project. To assist BWIP management in the evaluation process, the Systems Department staff has developed a BWIP Technical Program Evaluation Process (TPEP). The basic process relates progress on project technical work to the BWIP Functional and System Performance Criteria as defined in National Waste Terminal Storage (NWTS) Criteria Documents. The benefits of the TPEP to BWIP and future plans for TPEP are discussed. During fiscal year (FY) 1982, TPEP will be further formalized and further applied to the review of BWIP technical activities

  1. Basalt Waste Isolation Project Technical Program Evaluation Process: a criteria-based method

    International Nuclear Information System (INIS)

    Babad, H.; Evans, G.C.; Wolfe, B.A.

    1982-01-01

    The need to objectively evaluate the progress being made by the Basalt Waste Isolation Project (BWIP) toward establishing the feasibility of siting a nuclear waste repository in basalt (NWRB) mandates a process for evaluating the technical work of the project. To assist BWIP management in the evaluation process, the Systems Department staff has developed a BWIP Technical Program Evaluation Process (TPEP). The basic process relates progress on project technical work to the SWIP Functional and System Performance Criteria as defined in National Waste Terminal Storage (MWTS) Criteria Documents. The benefits of the TPEP to BWIP and future plans for TPEP are discussed. During fiscal year (FY) 1982, TPEP wll be further formalized and further applied to the review of BWIP technical activities

  2. Real-time PCR-based method for rapid detection of Aspergillus niger and Aspergillus welwitschiae isolated from coffee.

    Science.gov (United States)

    von Hertwig, Aline Morgan; Sant'Ana, Anderson S; Sartori, Daniele; da Silva, Josué José; Nascimento, Maristela S; Iamanaka, Beatriz Thie; Pelegrinelli Fungaro, Maria Helena; Taniwaki, Marta Hiromi

    2018-05-01

    Some species from Aspergillus section Nigri are morphologically very similar and altogether have been called A. niger aggregate. Although the species included in this group are morphologically very similar, they differ in their ability to produce mycotoxins and other metabolites and their taxonomical status has evolved continuously. Among them, A. niger and A. welwitschiae are ochratoxin A and fumonisin B 2 producers and their detection and/or identification is of crucial importance for food safety. The aim of this study was the development of a real-time PCR-based method for simultaneous discrimination of A. niger and A. welwitschiae from other species of the A. niger aggregate isolated from coffee beans. One primer pair and a hybridization probe specific for detection of A. niger and A. welwitschiae strains were designed based on the BenA gene sequences, and used in a Real-time PCR assay for the rapid discrimination between both these species from all others of the A. niger aggregate. The Real-time PCR assay was shown to be 100% efficient in discriminating the 73 isolates of A. niger/A. welwitschiae from the other A. niger aggregate species analyzed as a negative control. This result testifies to the use of this technique as a good tool in the rapid detection of these important toxigenic species. Copyright © 2018 Elsevier B.V. All rights reserved.

  3. Equilibration kinetics in isolated and membrane-bound photosynthetic reaction centers upon illumination: a method to determine the photoexcitation rate.

    Science.gov (United States)

    Manzo, Anthony J; Goushcha, Alexander O; Barabash, Yuri M; Kharkyanen, Valery N; Scott, Gary W

    2009-07-01

    Kinetics of electron transfer, following variation of actinic light intensity, for photosynthetic reaction centers (RCs) of purple bacteria (isolated and membrane-bound) were analyzed by measuring absorbance changes in the primary photoelectron donor absorption band at 865 nm. The bleaching of the primary photoelectron donor absorption band in RCs, following a sudden increase of illumination from the dark to an actinic light intensity of I(exp), obeys a simple exponential law with the rate constant alphaI(exp) + k(rec), in which alpha is a parameter relating the light intensity, measured in mW/cm(2), to a corresponding theoretical rate in units of reciprocal seconds, and k(rec) is the effective rate constant of the charge recombination in the photosynthetic RCs. In this work, a method for determining the alpha parameter value is developed and experimentally verified for isolated and membrane-bound RCs, allowing for rigorous modeling of RC macromolecule dynamics under varied photoexcitation conditions. Such modeling is necessary for RCs due to alterations of the forward photoexcitation rates and relaxation rates caused by illumination history and intramolecular structural dynamics effects. It is demonstrated that the classical Bouguer-Lambert-Beer formalism can be applied for the samples with relatively low scattering, which is not necessarily the case with strongly scattering media or high light intensity excitation.

  4. Using containerless methods to develop amorphous pharmaceuticals.

    Science.gov (United States)

    Weber, J K R; Benmore, C J; Suthar, K J; Tamalonis, A J; Alderman, O L G; Sendelbach, S; Kondev, V; Yarger, J; Rey, C A; Byrn, S R

    2017-01-01

    Many pipeline drugs have low solubility in their crystalline state and require compounding in special dosage forms to increase bioavailability for oral administration. The use of amorphous formulations increases solubility and uptake of active pharmaceutical ingredients. These forms are rapidly gaining commercial importance for both pre-clinical and clinical use. Synthesis of amorphous drugs was performed using an acoustic levitation containerless processing method and spray drying. The structure of the products was investigated using in-situ high energy X-ray diffraction. Selected solvents for processing drugs were investigated using acoustic levitation. The stability of amorphous samples was measured using X-ray diffraction. Samples processed using both spray drying and containerless synthesis were compared. We review methods for making amorphous pharmaceuticals and present data on materials made by containerless processing and spray drying. It was shown that containerless processing using acoustic levitation can be used to make phase-pure forms of drugs that are known to be difficult to amorphize. The stability and structure of the materials was investigated in the context of developing and making clinically useful formulations. Amorphous compounds are emerging as an important component of drug development and for the oral delivery of drugs with low solubility. Containerless techniques can be used to efficiently synthesize small quantities of pure amorphous forms that are potentially useful in pre-clinical trials and for use in the optimization of clinical products. Developing new pharmaceutical products is an essential enterprise to improve patient outcomes. The development and application of amorphous pharmaceuticals to increase absorption is rapidly gaining importance and it provides opportunities for breakthrough research on new drugs. There is an urgent need to solve problems associated with making formulations that are both stable and that provide high

  5. Development of numerical methods for reactive transport

    International Nuclear Information System (INIS)

    Bouillard, N.

    2006-12-01

    When a radioactive waste is stored in deep geological disposals, it is expected that the waste package will be damaged under water action (concrete leaching, iron corrosion). Then, to understand these damaging processes, chemical reactions and solutes transport are modelled. Numerical simulations of reactive transport can be done sequentially by the coupling of several codes. This is the case of the software platform ALLIANCES which is developed jointly with CEA, ANDRA and EDF. Stiff reactions like precipitation-dissolution are crucial for the radioactive waste storage applications, but standard sequential iterative approaches like Picard's fail in solving rapidly reactive transport simulations with such stiff reactions. In the first part of this work, we focus on a simplified precipitation and dissolution process: a system made up with one solid species and two aqueous species moving by diffusion is studied mathematically. It is assumed that a precipitation dissolution reaction occurs in between them, and it is modelled by a discontinuous kinetics law of unknown sign. By using monotonicity properties, the convergence of a finite volume scheme on admissible mesh is proved. Existence of a weak solution is obtained as a by-product of the convergence of the scheme. The second part is dedicated to coupling algorithms which improve Picard's method and can be easily used in an existing coupling code. By extending previous works, we propose a general and adaptable framework to solve nonlinear systems. Indeed by selecting special options, we can either recover well known methods, like nonlinear conjugate gradient methods, or design specific method. This algorithm has two main steps, a preconditioning one and an acceleration one. This algorithm is tested on several examples, some of them being rather academical and others being more realistic. We test it on the 'three species model'' example. Other reactive transport simulations use an external chemical code CHESS. For a

  6. Ex vivo piperaquine resistance developed rapidly in Plasmodium falciparum isolates in northern Cambodia compared to Thailand

    Directory of Open Access Journals (Sweden)

    Suwanna Chaorattanakawee

    2016-10-01

    Full Text Available Abstract Background The recent dramatic decline in dihydroartemisinin-piperaquine (DHA-PPQ efficacy in northwestern Cambodia has raised concerns about the rapid spread of piperaquine resistance just as DHA-PPQ is being introduced as first-line therapy in neighbouring countries. Methods Ex vivo parasite susceptibilities were tracked to determine the rate of progression of DHA, PPQ and mefloquine (MQ resistance from sentinel sites on the Thai–Cambodian and Thai–Myanmar borders from 2010 to 2015. Immediate ex vivo (IEV histidine-rich protein 2 (HRP-2 assays were used on fresh patient Plasmodium falciparum isolates to determine drug susceptibility profiles. Results IEV HRP-2 assays detected the precipitous emergence of PPQ resistance in Cambodia beginning in 2013 when 40 % of isolates had an IC90 greater than the upper limit of prior years, and this rate doubled to 80 % by 2015. In contrast, Thai–Myanmar isolates from 2013 to 14 remained PPQ-sensitive, while northeastern Thai isolates appeared to have an intermediate resistance profile. The opposite trend was observed for MQ where Cambodian isolates appeared to have a modest increase in overall sensitivity during the same period, with IC50 declining to median levels comparable to those found in Thailand. A significant association between increased PPQ IC50 and IC90 among Cambodian isolates with DHA-PPQ treatment failure was observed. Nearly all Cambodian and Thai isolates were deemed artemisinin resistant with a >1 % survival rate for DHA in the ring-stage assay (RSA, though there was no correlation among isolates to indicate cross-resistance between PPQ and artemisinins. Conclusions Clinical DHA-PPQ failures appear to be associated with declines in the long-acting partner drug PPQ, though sensitivity appears to remain largely intact for now in western Thailand. Rapid progression of PPQ resistance associated with DHA-PPQ treatment failures in northern Cambodia limits drugs of choice in

  7. Two efficient methods for isolation of high-quality genomic DNA from entomopathogenic fungi.

    Science.gov (United States)

    Serna-Domínguez, María G; Andrade-Michel, Gilda Y; Arredondo-Bernal, Hugo C; Gallou, Adrien

    2018-03-27

    Conventional and commercial methods for isolation of nucleic acids are available for fungal samples including entomopathogenic fungi (EPF). However, there is not a unique optimal method for all organisms. The cell wall structure and the wide range of secondary metabolites of EPF can broadly interfere with the efficiency of the DNA extraction protocol. This study compares three commercial protocols: DNeasy® Plant Mini Kit (Qiagen), Wizard® Genomic DNA Purification Kit (Promega), and Axygen™ Multisource Genomic DNA Miniprep Kit (Axygen) and three conventional methods based on different buffers: SDS, CTAB/PVPP, and CTAB/β-mercaptoethanol versus three cell lysis procedures: liquid nitrogen homogenization and two bead-beating materials (i.e., tungsten-carbide and stainless-steel) for four representative species of EPF (i.e., Beauveria bassiana, Hirsutella citriformis, Isaria javanica, and Metarhizium anisopliae). Liquid nitrogen homogenization combined with DNeasy® Plant Mini Kit (i.e., QN) or SDS buffer (i.e., SN) significantly improved the yield with a good purity (~1.8) and high integrity (>20,000 bp) of genomic DNA in contrast with other methods, also, these results were better when compared with the two bead-beating materials. The purified DNA was evaluated by PCR-based techniques: amplification of translation elongation factor 1-α (TEF) and two highly sensitive molecular markers (i.e., ISSR and AFLP) with reliable and reproducible results. Despite a variation in yield, purity, and integrity of extracted DNA across the four species of EPF with the different DNA extraction methods, the SN and QN protocols maintained a high-quality of DNA which is required for downstream molecular applications. Copyright © 2018 Elsevier B.V. All rights reserved.

  8. Development of radio-labeling method for natural juvenile hormone

    International Nuclear Information System (INIS)

    Kurata, Keiji; Shiozuki, Takahiro; Kotaki, Toyomi

    1997-01-01

    The aim of this study was to develop a new method for quantitative determination of juvenile hormone (JH) based on the principle for radioimmuno assay. Using JH-binding protein (JHBP), the discrimination of the L-form of JH from D-form not occurring naturally was attempted to establish a measuring method for JH. First, the corpus allatum, the JH-producing endocrine organ was cultured and both L-form JH and 3 H or 14 C-labelled JH could be obtained easily. There are several homologs of JH and it is necessary to establish the respective labelling methods for the homologues. Since different species of insects produce JH with its specific structure, each homologue could be produced by selecting an appropriate species. The capacity of JH production was compared among five insects. The biosynthetic ability by the corpus allatum from migratory locust was highest among them and 1 μg of labelled JH type 3 could be obtained from the culture with about 50 corpus allata for 3 hrs. Since other materials than JH were also released into the culture medium, establishment of the effective method for isolation and purification of JH is necessary to use it for bioassay. (M.N.)

  9. Rapid, sensitive and cost effective method for isolation of viral DNA from feacal samples of dogs

    Directory of Open Access Journals (Sweden)

    Savi.

    2010-06-01

    Full Text Available A simple method for viral DNA extraction using chelex resin was developed. The method used was eco-friendly and cost effective compared to other methods such as phenol chloroform method which use health hazardous organic reagents. Further, a polymerase chain reaction (PCR based detection of canine parvovirus (CPV using primers from conserved region of VP2 gene was developed. To increase the sensitivity and specificity of reaction, nested PCR was designed. PCR reaction was optimized to amplify 747bp product of VP2 gene. The assay can be completed in few hours and doesn’t need hazardous chemicals. Thus, the sample preparation using chelating resin along with nested PCR seems to be a sensitive, specific and practical method for the detection of CPV in diarrhoeal feacal samples. [Vet. World 2010; 3(3.000: 105-106

  10. Development and analysis of finite volume methods

    International Nuclear Information System (INIS)

    Omnes, P.

    2010-05-01

    This document is a synthesis of a set of works concerning the development and the analysis of finite volume methods used for the numerical approximation of partial differential equations (PDEs) stemming from physics. In the first part, the document deals with co-localized Godunov type schemes for the Maxwell and wave equations, with a study on the loss of precision of this scheme at low Mach number. In the second part, discrete differential operators are built on fairly general, in particular very distorted or nonconforming, bidimensional meshes. These operators are used to approach the solutions of PDEs modelling diffusion, electro and magneto-statics and electromagnetism by the discrete duality finite volume method (DDFV) on staggered meshes. The third part presents the numerical analysis and some a priori as well as a posteriori error estimations for the discretization of the Laplace equation by the DDFV scheme. The last part is devoted to the order of convergence in the L2 norm of the finite volume approximation of the solution of the Laplace equation in one dimension and on meshes with orthogonality properties in two dimensions. Necessary and sufficient conditions, relatively to the mesh geometry and to the regularity of the data, are provided that ensure the second-order convergence of the method. (author)

  11. Method development for trace and ultratrace analysis

    International Nuclear Information System (INIS)

    Anon.

    1992-01-01

    Method development, that is, selection of a mode of chromatography and the right column and mobile-phase combination for trace and ultratrace analysis, requires several main considerations. The method should be useful for resolving various trace and ultratrace components present in the sample. If the nature of these components is known, the choice of method may be straightforward, that is, a selection can be made from the following modes of HPLC: (1) adsorption chromatography; (2) normal-phase chromatography; (3) reversed-phase chromatography; (4) ion-pair chromatography; (5) ion-exchange chromatography; (6) ion chromatography. Unfortunately, the nature of all of the components is frequently unknown. However, several intelligent judgments can be made on the nature of impurities. This chapter deals with some basic approaches to mobile-phase selection and optimization. More detailed information may be found in basic texts. Techniques for separation of high-molecular-weight compounds (macromolecules) and chiral compounds may be found elsewhere. Mainly compounds with molecular weight lower than 2,000 are discussed here. 123 refs

  12. The development of permanent isolation surface barriers: Hanford Site, Richland, Washington, U.S.A

    International Nuclear Information System (INIS)

    Wing, N.R.; Gee, G.W.

    1993-01-01

    Permanent isolation surface barriers are being developed to isolate wastes disposed of in situ (in place) at the US Department of Energy's Hanford Site in Washington State (USA). The current focus of development efforts is to design barriers that will function in a semiarid to subhumid climate, Emit infiltration and percolation of water through the waste zone to near-zero amounts, be maintenance free, and last up to 1000 years or more. A series of field tests, experiments, and lysimeter studies have been conducted for several years. The results of tests to date confirm that the Hanford barrier concepts are valid for both present and wetter climatic conditions. The data collected also have provided the foundation for the design of a large prototype barrier to be constructed later in 1993. This paper presents the results of some of the field tests, experiments, and lysimeter studies

  13. Method of developing thick sloping coal formations

    Energy Technology Data Exchange (ETDEWEB)

    Bragintsev, V F; Mashkovtsev, I L; Semenov, V S; Zykov, V N

    1980-02-15

    A method of developing thick sloping coal formations in three inclined layers includes carrying out developmental operations for each of the layers until one begins development of the last one and extraction of layers. It is characterized in that in order to improve efficiency and safety of an operation of formation development there is first extraction of the upper layer and then slits in sequence from the roof of the formation to the floor of the upper layer and beneath protected objects. Then the lower layer is workedin thin strips in sequence from the floor of the formation to the roof of the lower layer. Next there is extraction of a slit at the roof of the middle layer and delivery of a plasticized hardening mixture into the worked out space of the indicated slot. The middle layer is worked in thin layers beneath the protection of the formed artificial roof in a sequence from the artificial roof to the floor of the middle layer. Workings of the middle layer are formed by joining of the combined workings of the upper and lower layers during extraction of pillars of coal between them. The layers are respectively worked following completion of roof advance in front of the working face of each subsequent extraction layer in alternating fashion.

  14. High Performance Harmonic Isolation By Means of The Single-phase Series Active Filter Employing The Waveform Reconstruction Method

    DEFF Research Database (Denmark)

    Senturk, Osman Selcuk; Hava, Ahmet M.

    2009-01-01

    current sampling delay reduction method (SDRM), a single-phase SAF compensated system provides higher harmonic isolation performance and higher stability margins compared to the system using conventional synchronous reference frame based methods. The analytical, simulation, and experimental studies of a 2...

  15. COMPARATIVE ANALYSIS OF METHODS FOR IDENTIFICATION OF NONTUBERCULOUS MYCOBACTERIA ISOLATED FROM CLINICAL MATERIAL

    Directory of Open Access Journals (Sweden)

    A. V. Lyamin

    2017-01-01

    Full Text Available Recently there has been a significant increase in the incidence of mycobacteriosis, which is due to an increase in the proportion of immunosuppressed patients, the presence of these various comorbid conditions, as well as the improvement of diagnostic methods. Selecting the most accurate method of identification is extremely important in determining treatment strategy of patients. The aim of the study was to conduct a comparative analysis of modern methods of identification NTMB isolated from clinical specimens in 2015 in the Samara region. The work was carried out identification of 78 strains of microorganisms. Laboratory diagnosis was carried out using the DNA hybridization method and MALDI-ToF mass spectrometry. When microbial identification using MALDI-ToF mass spectrometry was isolated 16 strains (20.5% M. kansasii; 11 strains (14.1% M. avium and M. fortuitum; 9 strains (11.5% M. gordonae; strain 3 (3.8% M. peregrinum, M. szulgai, M. chimera intracellulare group, strain 2 (2.6% M. abscessus, M. septicum, M. paragordonae, M. senegalence, 1 strain (1.3% M. chelonae, M. frederiksbergense, M. monacense, M. lentiflavum. By using mass spectrometry, it was identified 15 types NTMB compared with 9 types — by DNA hybridization. Full match identification results was observed only in 45 (57.7% strains of divergent strains were found in 16 (20.5%. Most often when using the DNA hybridization method, discrepancy was detected in slow-growing cultures (9 strains with a predominance of microorganisms identified as M. gordonae. Among the representatives of fast-growing NTMB, seven investigations were identified in the identification, more often among representatives of the M. fortuitum and M. peregrinum groups. Particular attention should be paid to the identification of the M. kansasii strain by a molecular genetic method, which mass spectrometry was defined as M. bovis. Both cultures of M. tuberculosis complex, which were identified by MALDI

  16. 105-KE Basin isolation barrier leak rate test analytical development. Revision 1

    International Nuclear Information System (INIS)

    Irwin, J.J.

    1995-01-01

    This document provides an analytical development in support of the proposed leak rate test of the 105-KE Basin. The analytical basis upon which the K-basin leak test results will be used to determine the basin leakage rates is developed in this report. The leakage of the K-Basin isolation barriers under postulated accident conditions will be determined from the test results. There are two fundamental flow regimes that may exist in the postulated K-Basin leakage: viscous laminar and turbulent flow. An analytical development is presented for each flow regime. The basic geometry and nomenclature of the postulated leak paths are denoted

  17. Calculating regional tissue volume for hyperthermic isolated limb perfusion: Four methods compared.

    Science.gov (United States)

    Cecchin, D; Negri, A; Frigo, A C; Bui, F; Zucchetta, P; Bodanza, V; Gregianin, M; Campana, L G; Rossi, C R; Rastrelli, M

    2016-12-01

    Hyperthermic isolated limb perfusion (HILP) can be performed as an alternative to amputation for soft tissue sarcomas and melanomas of the extremities. Melphalan and tumor necrosis factor-alpha are used at a dosage that depends on the volume of the limb. Regional tissue volume is traditionally measured for the purposes of HILP using water displacement volumetry (WDV). Although this technique is considered the gold standard, it is time-consuming and complicated to implement, especially in obese and elderly patients. The aim of the present study was to compare the different methods described in the literature for calculating regional tissue volume in the HILP setting, and to validate an open source software. We reviewed the charts of 22 patients (11 males and 11 females) who had non-disseminated melanoma with in-transit metastases or sarcoma of the lower limb. We calculated the volume of the limb using four different methods: WDV, tape measurements and segmentation of computed tomography images using Osirix and Oncentra Masterplan softwares. The overall comparison provided a concordance correlation coefficient (CCC) of 0.92 for the calculations of whole limb volume. In particular, when Osirix was compared with Oncentra (validated for volume measures and used in radiotherapy), the concordance was near-perfect for the calculation of the whole limb volume (CCC = 0.99). With methods based on CT the user can choose a reliable plane for segmentation purposes. CT-based methods also provides the opportunity to separate the whole limb volume into defined tissue volumes (cortical bone, fat and water). Copyright © 2016 Elsevier Ltd. All rights reserved.

  18. Filtration Isolation of Nucleic Acids: A Simple and Rapid DNA Extraction Method.

    Science.gov (United States)

    McFall, Sally M; Neto, Mário F; Reed, Jennifer L; Wagner, Robin L

    2016-08-06

    FINA, filtration isolation of nucleic acids, is a novel extraction method which utilizes vertical filtration via a separation membrane and absorbent pad to extract cellular DNA from whole blood in less than 2 min. The blood specimen is treated with detergent, mixed briefly and applied by pipet to the separation membrane. The lysate wicks into the blotting pad due to capillary action, capturing the genomic DNA on the surface of the separation membrane. The extracted DNA is retained on the membrane during a simple wash step wherein PCR inhibitors are wicked into the absorbent blotting pad. The membrane containing the entrapped DNA is then added to the PCR reaction without further purification. This simple method does not require laboratory equipment and can be easily implemented with inexpensive laboratory supplies. Here we describe a protocol for highly sensitive detection and quantitation of HIV-1 proviral DNA from 100 µl whole blood as a model for early infant diagnosis of HIV that could readily be adapted to other genetic targets.

  19. Vibration isolation design for periodically stiffened shells by the wave finite element method

    Science.gov (United States)

    Hong, Jie; He, Xueqing; Zhang, Dayi; Zhang, Bing; Ma, Yanhong

    2018-04-01

    Periodically stiffened shell structures are widely used due to their excellent specific strength, in particular for aeronautical and astronautical components. This paper presents an improved Wave Finite Element Method (FEM) that can be employed to predict the band-gap characteristics of stiffened shell structures efficiently. An aero-engine casing, which is a typical periodically stiffened shell structure, was employed to verify the validation and efficiency of the Wave FEM. Good agreement has been found between the Wave FEM and the classical FEM for different boundary conditions. One effective wave selection method based on the Wave FEM has thus been put forward to filter the radial modes of a shell structure. Furthermore, an optimisation strategy by the combination of the Wave FEM and genetic algorithm was presented for periodically stiffened shell structures. The optimal out-of-plane band gap and the mass of the whole structure can be achieved by the optimisation strategy under an aerodynamic load. Results also indicate that geometric parameters of stiffeners can be properly selected that the out-of-plane vibration attenuates significantly in the frequency band of interest. This study can provide valuable references for designing the band gaps of vibration isolation.

  20. Comparison of Composition and Anticaries Effect of Galla Chinensis Extracts with Different Isolation Methods

    Science.gov (United States)

    Huang, Xuelian; Deng, Meng; Liu, Mingdong; Cheng, Lei; Exterkate, R.A.M.; Li, Jiyao; Zhou, Xuedong; Ten Cate, Jacob. M.

    2017-01-01

    Objectives: Galla chinensis water extract (GCE) has been demonstrated to inhibit dental caries by favorably shifting the demineralization/remineralization balance of enamel and inhibiting the biomass and acid formation of dental biofilm. The present study focused on the comparison of composition and anticaries effect of Galla chinensis extracts with different isolation methods, aiming to improve the efficacy of caries prevention. Methods: The composition of water extract (GCE), ethanol extract (eGCE) and commercial tannic acid was compared. High performance liquid chromatography coupled to electrospray ionization-time of flight-mass spectrometry (HPLC-ESI-TOF-MS) analysis was used to analyze the main ingredients. In vitro pH-cycling regime and polymicrobial biofilms model were used to assess the ability of different Galla chinensis extracts to inhibit enamel demineralization, acid formation and biofilm formation. Results: All the GCE, eGCE and tannic acid contained a high level of total phenolics. HPLC-ESI-TOF-MS analysis showed that the main ingredients of GCE were gallic acid (GA), while eGCE mainly contained 4-7 galloylglucopyranoses (GGs) and tannic acid mainly contained 5-10 GGs. Furthermore, eGCE and tannic acid showed a better effect on inhibiting enamel demineralization, acid formation and biofilm formation compared to GCE. Conclusions: Galla chinensis extracts with higher tannin content were suggested to have higher potential to prevent dental caries. PMID:28979574

  1. XRSW method, its application and development

    Energy Technology Data Exchange (ETDEWEB)

    Zheludeva, S I; Kovalchuk, M V [Russian Academy of Sciences, Institute of Crystallography, Moscow (Russian Federation)

    1996-09-01

    X-Ray Standing Waves (XRSW) may be obtained under dynamical diffraction in periodic structures or under total external reflection conditions (TR) is stratified medium. As the incident angle varies, XRSW nodes and antinodes move in the direction perpendicular to the reflecting planes, leading to drastic variation of photoelectron interaction of X-ray with matter and resulting in specific angular dependencies of secondary radiation yields (photoelectrons, fluorescence, internal photoeffect, photoluminescence, Compton and thermal diffuse scattering). The structural information - the position of investigated atoms in the direction of XRSW movement (coherent position), the distribution of atoms about this position (coherent fraction) - is obtained with the accuracy about several percents from XRSW period D. The objects under investigation are: semiconductor surface layers, heterostructure, multicomponent crystals, interfaces, adsorbed layers. Besides the development of XRSW method allow to obtain structure, geometrical and optical parameters of ultrathin films (crystalline and disordered, organic and inorganic) and nanostructures on their base.

  2. Effect of drying methods on the structure, thermo and functional properties of fenugreek (Trigonella foenum graecum) protein isolate.

    Science.gov (United States)

    Feyzi, Samira; Varidi, Mehdi; Zare, Fatemeh; Varidi, Mohammad Javad

    2018-03-01

    Different drying methods due to protein denaturation could alter the functional properties of proteins, as well as their structure. So, this study focused on the effect of different drying methods on amino acid content, thermo and functional properties, and protein structure of fenugreek protein isolate. Freeze and spray drying methods resulted in comparable protein solubility, dynamic surface and interfacial tensions, foaming and emulsifying properties except for emulsion stability. Vacuum oven drying promoted emulsion stability, surface hydrophobicity and viscosity of fenugreek protein isolate at the expanse of its protein solubility. Vacuum oven process caused a higher level of Maillard reaction followed by the spray drying process, which was confirmed by the lower amount of lysine content and less lightness, also more browning intensity. ΔH of fenugreek protein isolates was higher than soy protein isolate, which confirmed the presence of more ordered structures. Also, the bands which are attributed to the α-helix structures in the FTIR spectrum were in the shorter wave number region for freeze and spray dried fenugreek protein isolates that show more possibility of such structures. This research suggests that any drying method must be conducted in its gentle state in order to sustain native structure of proteins and promote their functionalities. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  3. A Method for Isolation of Pasteuria penetrans Endospores for Bioassay and Genomic Studies.

    Science.gov (United States)

    Waterman, Jenora T; Bird, David McK; Opperman, Charles H

    2006-03-01

    A rapid method for collection of Pasteuria penetrans endospores was developed. Roots containing P. penetrans-infected root-knot nematode females were softened by pectinase digestion, mechanically processed, and filtered to collect large numbers of viable endospores. This method obviates laborious handpicking of Pasteuria-infected females and yields endospores competent to attach to and infect nematodes. Endospores are suitable for morphology studies and DNA preparations.

  4. Development of a Radial Deconsolidation Method

    Energy Technology Data Exchange (ETDEWEB)

    Helmreich, Grant W. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Montgomery, Fred C. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Hunn, John D. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)

    2015-12-01

    A series of experiments have been initiated to determine the retention or mobility of fission products* in AGR fuel compacts [Petti, et al. 2010]. This information is needed to refine fission product transport models. The AGR-3/4 irradiation test involved half-inch-long compacts that each contained twenty designed-to-fail (DTF) particles, with 20-μm thick carbon-coated kernels whose coatings were deliberately fabricated such that they would crack under irradiation, providing a known source of post-irradiation isotopes. The DTF particles in these compacts were axially distributed along the compact centerline so that the diffusion of fission products released from the DTF kernels would be radially symmetric [Hunn, et al. 2012; Hunn et al. 2011; Kercher, et al. 2011; Hunn, et al. 2007]. Compacts containing DTF particles were irradiated at Idaho National Laboratory (INL) at the Advanced Test Reactor (ATR) [Collin, 2015]. Analysis of the diffusion of these various post-irradiation isotopes through the compact requires a method to radially deconsolidate the compacts so that nested-annular volumes may be analyzed for post-irradiation isotope inventory in the compact matrix, TRISO outer pyrolytic carbon (OPyC), and DTF kernels. An effective radial deconsolidation method and apparatus appropriate to this application has been developed and parametrically characterized.

  5. Method Development to Increase Protein Enrichment During Dry Fractionation of Starch-Rich Legumes

    NARCIS (Netherlands)

    Pelgrom, P.J.M.; Boom, R.M.; Schutyser, M.A.I.

    2015-01-01

    A facile method was developed to establish milling settings that optimally separate starch granules from protein bodies and cell wall fibres for starch-rich legumes. Optimal separation was obtained for pea, bean, lentil and chickpea when the particle size distribution curve of flour and isolated

  6. DEVELOPMENT OF A MOLECULAR METHOD TO IDENTIFY HEPATITIS E VIRUS IN WATER

    Science.gov (United States)

    Hepatitis E virus (HEV) causes an infectious form of hepatitis associated with contaminated water. By analyzing the sequence of several HEV isolates, a reverse transciption-polymerase chain reaction method was developed and optimized that should be able to identify all of the kn...

  7. A simple electroelution method for rapid protein purification: isolation and antibody production of alpha toxin from Clostridium septicum

    Directory of Open Access Journals (Sweden)

    Lorena Vázquez-Iglesias

    2017-06-01

    Full Text Available Clostridium septicum produces a number of diseases in human and farm animals which, in most of the cases, are fatal without clinical intervention. Alpha toxin is an important agent and the unique lethal virulent factor produced by Clostridium septicum. This toxin is haemolytic, highly lethal and necrotizing activities but is being used as an antigen to develop animal vaccines. The aim of this study was to isolate the alpha toxin of Clostridium septicum and produce highly specific antibodies against it. In this work, we have developed a simple and efficient method for alpha toxin purification, based on electroelution that can be used as a time-saving method for purifying proteins. This technique avoids contamination by other proteins that could appear during other protein purification techniques such chromatography. The highly purified toxin was used to produce polyclonal antibodies. The specificity of the antibodies was tested by western blot and these antibodies can be applied to the quantitative determination of alpha toxin by slot blot.

  8. Development of methods for evaluating active faults

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    2013-08-15

    The report for long-term evaluation of active faults was published by the Headquarters for Earthquake Research Promotion on Nov. 2010. After occurrence of the 2011 Tohoku-oki earthquake, the safety review guide with regard to geology and ground of site was revised by the Nuclear Safety Commission on Mar. 2012 with scientific knowledges of the earthquake. The Nuclear Regulation Authority established on Sep. 2012 is newly planning the New Safety Design Standard related to Earthquakes and Tsunamis of Light Water Nuclear Power Reactor Facilities. With respect to those guides and standards, our investigations for developing the methods of evaluating active faults are as follows; (1) For better evaluation on activities of offshore fault, we proposed a work flow to date marine terrace (indicator for offshore fault activity) during the last 400,000 years. We also developed the analysis of fault-related fold for evaluating of blind fault. (2) To clarify the activities of active faults without superstratum, we carried out the color analysis of fault gouge and divided the activities into thousand of years and tens of thousands. (3) To reduce uncertainties of fault activities and frequency of earthquakes, we compiled the survey data and possible errors. (4) For improving seismic hazard analysis, we compiled the fault activities of the Yunotake and Itozawa faults, induced by the 2011 Tohoku-oki earthquake. (author)

  9. The development of interdepartmental audit methods

    International Nuclear Information System (INIS)

    Thwaites, D.I.; Allahverdi, M.

    1995-01-01

    The UK Radiotherapy Physics Audit Network is now well-established, with seven network groups and co-ordinated by the IPSM. It is based on visits, using ion chambers as the measurement method, and auditing at least machine calibration, single field parameters and simple multi-field planned irradiations. In addition procedural audit of dosimetry and quality control procedures, and records is incorporated. The general approach has been to use interdepartmental audit involving mutual co-operation with peer professionals from other centres. The different groups have evolved at different paces and in rather different directions. However the IPSM coordinating role ensures a basic common minimum to the system. The Scottish+ group has developed a semi-anatomical phantom to use in audit stages following on from the basic single field and geometric phantom dosimetry audit levels. This has been evaluated experimentally in one department before wider use. The Scottish+ audit system is briefly described. Results from levels 1 and 2 are summarised and the design and testing of the semi-anatomical phantom are discussed. The current and future development of the audit system is presented

  10. Method of developing thick sloping coal formations

    Energy Technology Data Exchange (ETDEWEB)

    Bragintsev, V F; Mashkovtsev, I L; Semenov, V S; Zykov, V N

    1980-04-25

    A method is patented for developing thick sloping coal formations in 3 inclined layers. It includes conducting developmental operations for each of the layers until one begins the last one and extraction of the layers. In order to improve effectivess and extraction operation safety one first carried out preliminary development of a formation in thin strips beneath protected objects when extracting formation which contain alot of gas. Then one removes the gas of a formation through boreholes that have been drilled into the formation from the indicated workings. Then one works the upper layer in thin strips in a sequence from the roof of the formation to the floor of the upper layer. The one strengthens roof rock of the formation by pumping in a quickly hardening solution into the boreholes which have been drilled into the roof of the formation after processing the upper layer. The middle layer is worked in thin strips in the sequence from the roof to the ground of the middle layer, then the lower layer of the formation is strengthened by pumping in quickly hardening solution into the formation along degasified boreholes and it is worked in thin strips in sequence from the ground of the lower to its roof. Workings are shaped respectively for the middle and lower layers by deepening workings of the upper and middle layers. The layers are worked respectively after finishing displacement of the roof in front of the extraction face of each subsequent extraction of a layer in alternating fashion.

  11. Development of a forward genetic screen to isolate oil mutants in the green microalga Chlamydomonas reinhardtii.

    Science.gov (United States)

    Cagnon, Caroline; Mirabella, Boris; Nguyen, Hoa Mai; Beyly-Adriano, Audrey; Bouvet, Séverine; Cuiné, Stéphan; Beisson, Fred; Peltier, Gilles; Li-Beisson, Yonghua

    2013-12-02

    Oils produced by microalgae are precursors to biodiesel. To achieve a profitable production of biodiesel from microalgae, identification of factors governing oil synthesis and turnover is desirable. The green microalga Chlamydomonas reinhardtii is amenable to genetic analyses and has recently emerged as a model to study oil metabolism. However, a detailed method to isolate various types of oil mutants that is adapted to Chlamydomonas has not been reported. We describe here a forward genetic approach to isolate mutants altered in oil synthesis and turnover from C. reinhardtii. It consists of a three-step screening procedure: a primary screen by flow cytometry of Nile red stained transformants grown in 96-deep-well plates under three sequential conditions (presence of nitrogen, then absence of nitrogen, followed by oil remobilization); a confirmation step using Nile red stained biological triplicates; and a validation step consisting of the quantification by thin layer chromatography of oil content of selected strains. Thirty-one mutants were isolated by screening 1,800 transformants generated by random insertional mutagenesis (1.7%). Five showed increased oil accumulation under the nitrogen-replete condition and 13 had altered oil content under nitrogen-depletion. All mutants were affected in oil remobilization. This study demonstrates that various types of oil mutants can be isolated in Chlamydomonas based on the method set-up here, including mutants accumulating oil under optimal biomass growth. The strategy conceived and the protocol set-up should be applicable to other microalgal species such as Nannochloropsis and Chlorella, thus serving as a useful tool in Chlamydomonas oil research and algal biotechnology.

  12. [Comparison of manual and automated (MagNA Pure) nucleic acid isolation methods in molecular diagnosis of HIV infections].

    Science.gov (United States)

    Alp, Alpaslan; Us, Dürdal; Hasçelik, Gülşen

    2004-01-01

    Rapid quantitative molecular methods are very important for the diagnosis of human immunodeficiency virus (HIV) infections, assessment of prognosis and follow up. The purpose of this study was to compare and evaluate the performances of conventional manual extraction method and automated MagNA Pure system, for the nucleic acid isolation step which is the first and most important step in molecular diagnosis of HIV infections. Plasma samples of 35 patients in which anti-HIV antibodies were found as positive by microparticule enzyme immunoassay and confirmed by immunoblotting method, were included in the study. The nucleic acids obtained simultaneously by manual isolation kit (Cobas Amplicor, HIV-1 Monitor Test, version 1.5, Roche Diagnostics) and automated system (MagNA Pure LC Total Nucleic Acid Isolation Kit, Roche Diagnostics), were amplified and detected in Cobas Amplicor (Roche Diagnostics) instrument. Twenty three of 35 samples (65.7%) were found to be positive, and 9 (25.7%) were negative by both of the methods. The agreement between the methods were detected as 91.4%, for qualitative results. Viral RNA copies detected by manual and MagNA Pure isolation methods were found between 76.0-7.590.000 (mean: 487.143) and 113.0-20.300.0000 (mean: 2.174.097) copies/ml, respectively. When both of the overall and individual results were evaluated, the number of RNA copies obtained with automatized system, were found higher than the manual method (p<0.05). Three samples which had low numbers of nucleic acids (113, 773, 857, respectively) with MagNA Pure, yielded negative results with manual method. In conclusion, the automatized MagNA Pure system was found to be a reliable, rapid and practical method for the isolation of HIV-RNA.

  13. Mapping isolated wetlands in a Karst landscape: GIS and remote sensing methods

    Science.gov (United States)

    Isolated wetlands occur in many areas of the United States, and although they are relatively common, they are a resource not yet thoroughly understood by the scientific community. Isolated wetlands have received increased attention recently, due to the 2001 Solid Waste Agency of ...

  14. Scope and status of Russian contribution for analysis methods for seismically isolated nuclear structure

    International Nuclear Information System (INIS)

    Beliayev, V.S.; Vinogradov, V.V.; Guskov, V.D.

    1993-01-01

    In the last few years, we can see in Russia the amplification of interest to problems of seismic isolation for potentially dangerous objects as the most effective way to alleviate the possible damage. This material comprises the data which characterize the level of theoretical design and experimental studying of seismic isolation systems of NPP components and structures. (author)

  15. Comparison of different methods of RNA isolation for plum pox virus detection by reverse transcription-polymerase chain reaction.

    Science.gov (United States)

    Faggioli, F; Pasquini, G; Barba, M

    1998-09-01

    The diagnosis of plum pox virus (PPV) is still considered one of the most important aspects of the "sharka" problem. In fact, different studies demonstrated an uneven distribution of the virus in infected trees due to a high variability in virus concentration. These aspects complicate the PPV diagnosis. To date, biological, serological and molecular assays have been successively developed in order to obtain sensitive and efficient PPV detection techniques. In particular, the polymerase chain reaction (PCR) technique seems to be promising and can be considered the most sensitive and reliable one. Preparation of viral RNA is still a fundamental step in reverse transcription-PCR (RT-PCR) technique, especially when applied to large scale testing, i.e., for certification purposes. In order to find the most rapid and efficient procedure, we have compared three different procedures of extraction of viral RNA to be processed RT-PCR. Their common characteristics is their capacity to extract the RNA from a small amount of plant tissue without organic solvents in the extraction fluid. The procedures were as follows: an immuno-capture (IC) method using a specific antiserum, a silica-capture (SC) method using a non-specific matrix, and a simple and rapid RNA extraction (RE) method. They all were followed by one-tube RT-PCR. The obtained results show that all the three techniques allowed a successful amplification and detection of PPV in tested samples except the SC-PCR method which proved less effective. In fact, the IC-PCR and RE-PCR methods amplified and detected PPV in all isolates tested, while the SC-PCR method was able to reveal the presence of the virus in apricot and infected control samples only.

  16. A high-throughput splinkerette-PCR method for the isolation and sequencing of retroviral insertion sites

    DEFF Research Database (Denmark)

    Uren, Anthony G; Mikkers, Harald; Kool, Jaap

    2009-01-01

    sites has been a major limitation to performing screens on this scale. Here we present a method for the high-throughput isolation of insertion sites using a highly efficient splinkerette-PCR method coupled with capillary or 454 sequencing. This protocol includes a description of the procedure for DNA......Insertional mutagens such as viruses and transposons are a useful tool for performing forward genetic screens in mice to discover cancer genes. These screens are most effective when performed using hundreds of mice; however, until recently, the cost-effective isolation and sequencing of insertion...

  17. Characterization of ß-Glucans Isolated from Brewer’s Yeast and Dried by Different Methods

    Directory of Open Access Journals (Sweden)

    Vesna Zechner-Krpan

    2010-01-01

    Full Text Available Two different procedures have been used for isolation of water-insoluble ß-glucans from brewer’s yeast: alkaline-acidic isolation (AA and alkaline-acidic isolation with mannoprotein removal (AAM. The obtained ß-glucans were then dried by air-drying, lyophilization and combination of sonication and spray-drying. ß-Glucan preparations obtained by AA and AAM isolations had similar values of dry mass, total polysaccharides, proteins and organic elemental microanalysis. The mass fractions of ß-glucan in total polysaccharides were significantly affected by different isolation procedures. Fourier transform infrared (FTIR spectra of all preparations had the appearance typical for (1→3-ß-D-glucan. Lyophilization and especially air-drying caused a higher degree of agglomeration and changes in ß-glucan microstructure. Sonication followed by spray-drying resulted in minimal structural changes and negligible formation of agglomerates.

  18. Chlorophyll biosynthesis and assembly into chlorophyll-protein complexes in isolated developing chloroplasts

    International Nuclear Information System (INIS)

    Bhaya, D.; Castelfranco, P.A.

    1985-01-01

    Isolated developing plastids from greening cucumber cotyledons or from photoperiodically grown pea seedlings incorporated 14 C-labeled 5-aminolevulinic acid (ALA) into chlorophyll (Chl). Incorporation was light dependent, enhanced by S-adenosylmethionine, and linear for 1 hr. The in vitro rate of Chl synthesis from ALA was comparable to the in vivo rate of Chl accumulation. Levulinic acid and dioxoheptanoic acid strongly inhibited Chl synthesis but not plastid protein synthesis. Neither chloramphenicol nor spectinomycin affected Chl synthesis, although protein synthesis was strongly inhibited. Components of thylakoid membranes from plastids incubated with [ 14 C]ALA were resolved by electrophoresis and then subjected to autoradiography. This work showed that (i) newly synthesized Chl was assembled into Chl-protein complexes and (ii) the inhibition of protein synthesis during the incubation did not alter the labeling pattern. Thus, there was no observable short-term coregulation between Chl synthesis (from ALA) and the synthesis of membrane proteins in isolated plastids

  19. Simulant Development for Hanford Tank Farms Double Valve Isolation (DVI) Valves Testing

    Energy Technology Data Exchange (ETDEWEB)

    Wells, Beric E.

    2012-12-21

    Leakage testing of a representative sample of the safety-significant isolation valves for Double Valve Isolation (DVI) in an environment that simulates the abrasive characteristics of the Hanford Tank Farms Waste Transfer System during waste feed delivery to the Waste Treatment and Immobilization Plant (WTP) is to be conducted. The testing will consist of periodic leak performed on the DVI valves after prescribed numbers of valve cycles (open and close) in a simulated environment representative of the abrasive properties of the waste and the Waste Transfer System. The valve operations include exposure to cycling conditions that include gravity drain and flush operation following slurry transfer. The simulant test will establish the performance characteristics and verify compliance with the Documented Safety Analysis. Proper simulant development is essential to ensure that the critical process streams characteristics are represented, National Research Council report “Advice on the Department of Energy's Cleanup Technology Roadmap: Gaps and Bridges”

  20. Recent developments of target and ion sources to produce ISOL beams

    CERN Document Server

    Stora, Thierry

    2013-01-01

    In this review on target and ion sources for ISOL (Isotope Separation OnLine) beams, important develop- ments from the past five years are highlighted. While at precedent EMIS conferences, a particular focus was given to a single topics, for instance specifically on ion sources or on chemical purification tech- niques, here each of the important elements present in an ISOL production unit is discussed. Fast diffus- ing nanomaterials, uranium-based targets, high power targets for next generation facilities, purification by selective adsorption, new ion sources are all part of this review. For each of these selected topics, the reported results lead to significant gains in intensity, purity, or quality of the delivered beam, or in the production of new isotope beams. Often the outcome resulted from the combination of original ideas with state-of-the-art investigations; this was carried out using very different scientific disciplines, lead- ing to understanding of the underlying chemical or physical mechanisms a...

  1. Whole Genome Sequencing of Enterovirus species C Isolates by High-throughput Sequencing: Development of Generic Primers

    Directory of Open Access Journals (Sweden)

    Maël Bessaud

    2016-08-01

    Full Text Available Enteroviruses are among the most common viruses infecting humans and can cause diverse clinical syndromes ranging from minor febrile illness to severe and potentially fatal diseases. Enterovirus species C (EV-C consists of more than 20 types, among which the 3 serotypes of polioviruses, the etiological agents of poliomyelitis, are included. Biodiversity and evolution of EV-C genomes are shaped by frequent recombination events. Therefore, identification and characterization of circulating EV-C strains require the sequencing of different genomic regions.A simple method was developed to sequence quickly the entire genome of EV-C isolates. Four overlapping fragments were produced separately by RT-PCR performed with generic primers. The four amplicons were then pooled and purified prior to be sequenced by high-throughput technique.The method was assessed on a panel of EV-Cs belonging to a wide-range of types. It can be used to determine full-length genome sequences through de novo assembly of thousands of reads. It was also able to discriminate reads from closely related viruses in mixtures.By decreasing the workload compared to classical Sanger-based techniques, this method will serve as a precious tool for sequencing large panels of EV-Cs isolated in cell cultures during environmental surveillance or from patients, including vaccine-derived polioviruses.

  2. Properties of nanocellulose isolated from corncob residue using sulfuric acid, formic acid, oxidative and mechanical methods.

    Science.gov (United States)

    Liu, Chao; Li, Bin; Du, Haishun; Lv, Dong; Zhang, Yuedong; Yu, Guang; Mu, Xindong; Peng, Hui

    2016-10-20

    In this work, nanocellulose was extracted from bleached corncob residue (CCR), an underutilized lignocellulose waste from furfural industry, using four different methods (i.e. sulfuric acid hydrolysis, formic acid (FA) hydrolysis, 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO)-mediated oxidation, and pulp refining, respectively). The self-assembled structure, morphology, dimension, crystallinity, chemical structure and thermal stability of prepared nanocellulose were investigated. FA hydrolysis produced longer cellulose nanocrystals (CNCs) than the one obtained by sulfuric acid hydrolysis, and resulted in high crystallinity and thermal stability due to its preferential degradation of amorphous cellulose and lignin. The cellulose nanofibrils (CNFs) with fine and individualized structure could be isolated by TEMPO-mediated oxidation. In comparison with other nanocellulose products, the intensive pulp refining led to the CNFs with the longest length and the thickest diameter. This comparative study can help to provide an insight into the utilization of CCR as a potential source for nanocellulose production. Copyright © 2016 Elsevier Ltd. All rights reserved.

  3. Various methods for isolation of multipotent human periodontal ligament cells for regenerative medicine.

    Science.gov (United States)

    Tran, Ha Le Bao; Doan, Vu Nguyen; Le, Huong Thi Ngoc; Ngo, Lan Thi Quynh

    2014-08-01

    Periodontal ligament (PDL) is a specialized connective tissue that connects cementum and alveolar bone to maintain and support the teeth in situ and preserve tissue homeostasis. Recent studies have revealed the existence of stem cells in human dental tissues including periodontal ligament that play an important role, not only in the maintenance of the periodontium but also in promoting periodontal regeneration. In this study, human periodontal ligament cells (hPDLCs) were isolated by outgrowth and enzymatic dissociation methods. Expression of surface markers on PDLCs as human mesenchymal stem cells (MSCs) was identified by flow cytometry. In addition, proliferation and differentiation capacity of cultured cells to osteoblasts, adipocytes were evaluated. As a result, we successfully cultured cells from the human periodontal ligament tissues. PDLCs express mesenchymal stem cell (MSC) markers such as CD44, CD73, and CD90 and do not express CD34, CD45, and HLA-DR. PDLCs also possess the multipotential to differentiate into various types of cells, such as osteoblast and adipocytes, in vitro. Therefore, these cells have high potential to serve as materials for tissue engineering, especially dental tissue engineering.

  4. Comparison of Methods for Isolating High Quality DNA and RNA from an Oleaginous Fungus Cunninghamella bainieri Strain 2a1

    Directory of Open Access Journals (Sweden)

    Noor Adila, A. K.

    2007-01-01

    Full Text Available A number of protocols have been reported for efficient fungal DNA and RNA isolation. However, many of these methods are often designed for certain groups or morphological forms of fungi and, in some cases, are species dependent. In this report, we compared four published protocols for DNA isolation from a locally isolated oleaginous fungus, Cunninghamella bainieri strain 2a1. These protocols either involved the use of polyvinyl pyrrolidone (PVP, hexacetyltrimethylammonium bromide (CTAB or without using PVB or CTAB. For RNA isolation, we tested two published protocols, one of which is based on TRI REAGENT (Molecular Research Center, USA and another is simple method employing phenol for RNA extraction and LiCl for precipitation. We found that the protocol involving the use of CTAB produced the highest genomic DNA yield with the best quality compared to other protocols. In the presence of CTAB, unwanted polysaccharides were removed and this method yielded an average amount of 816 ± 12.2 µg DNA/g mycelia with UV absorbance ratios A260/280 and A260/230 of 1.67 ± 0.64 and 1.97 ± 0.23, respectively. The genomic DNA isolated via this protocol is also suitable for PCR amplification and restriction enzyme digestion. As for RNA isolation, the method involving phenol extraction and LiCl precipitation produced the highest yield of RNA with an average amount of 372 ± 6.0 µg RNA/g mycelia. The RNA appears to be relatively pure since it has UV absorbance ratios A260/280 and A260/230 of 1.89 ± 2.00 and 1.99 ± 0.03, respectively. Finally, we have demonstrated that this method could produce RNA of sufficient quality for RT-PCR that amplified a 600 bp fragment of ∆12-fatty acid desaturase gene in C. bainieri.

  5. Social isolation and loneliness in later life: A parallel convergent mixed-methods case study of older adults and their residential contexts in the Minneapolis metropolitan area, USA.

    Science.gov (United States)

    Finlay, Jessica M; Kobayashi, Lindsay C

    2018-07-01

    Social isolation and loneliness are increasingly prevalent among older adults in the United States, with implications for morbidity and mortality risk. Little research to date has examined the complex person-place transactions that contribute to social well-being in later life. This study aimed to characterize personal and neighborhood contextual influences on social isolation and loneliness among older adults. Interviews were conducted with independent-dwelling men and women (n = 124; mean age 71 years) in the Minneapolis metropolitan area (USA) from June to October, 2015. A convergent mixed-methods design was applied, whereby quantitative and qualitative approaches were used in parallel to gain simultaneous insights into statistical associations and in-depth individual perspectives. Logistic regression models predicted self-reported social isolation and loneliness, adjusted for age, gender, past occupation, race/ethnicity, living alone, street type, residential location, and residential density. Qualitative thematic analyses of interview transcripts probed individual experiences with social isolation and loneliness. The quantitative results suggested that African American adults, those with a higher socioeconomic status, those who did not live alone, and those who lived closer to the city center were less likely to report feeling socially isolated or lonely. The qualitative results identified and explained variation in outcomes within each of these factors. They provided insight on those who lived alone but did not report feeling lonely, finding that solitude was sought after and enjoyed by a portion of participants. Poor physical and mental health often resulted in reporting social isolation, particularly when coupled with poor weather or low-density neighborhoods. At the same time, poor health sometimes provided opportunities for valued social engagement with caregivers, family, and friends. The combination of group-level risk factors and in-depth personal

  6. Development of methods for evaluating active faults

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    2012-08-15

    The HERP report for long-term evaluation of active faults and the NSC safety review guide with regard to geology and ground of site were published on Nov. 2010 and on Dec. 2010, respectively. With respect to those reports, our investigation is as follows; (1) For assessment of seismic hazard, we estimated seismic sources around NPPs based on information of tectonic geomorphology, earthquake distribution and subsurface geology. (2) For evaluation on the activity of blind fault, we calculated the slip rate on the 2008 Iwate-Miyagi Nairiku Earthquake fault, using information on late Quaternary fluvial terraces. (3) To evaluate the magnitude of earthquakes whose sources are difficult to identify, we proposed a new method for calculation of the seismogenic layer thickness. (4) To clarify the activities of active faults without superstratum, we carried out the color analysis of fault gouge and divided the activities into thousand of years and tens of thousands. (5) For improving chronology of sediments, we detected new widespread cryptotephras using mineral chemistry and developed late Quaternary cryptotephrostratigraphy around NPPs. (author)

  7. Developments of an Interactive Sail Design Method

    Directory of Open Access Journals (Sweden)

    S. M. Malpede

    2000-01-01

    Full Text Available This paper presents a new tool for performing the integrated design and analysis of a sail. The features of the system are the geometrical definition of a sail shape, using the Bezier surface method, the creation of a finite element model for the non-linear structural analysis and a fluid-dynamic model for the aerodynamic analysis. The system has been developed using MATLAB(r. Recent sail design efforts have been focused on solving the aeroelastic behavior of the sail. The pressure distribution on a sail changes continuously, by virtue of cloth stretch and flexing. The sail shape determines the pressure distribution and, at the same time, the pressure distribution on the sail stretches and flexes the sail material determining its shape. This characteristic non-linear behavior requires iterative solution strategies to obtain the equilibrium configuration and evaluate the forces involved. The aeroelastic problem is tackled by combining structural with aerodynamic analysis. Firstly, pressure loads for a known sail-shape are computed (aerodynamic analysis. Secondly, the sail-shape is analyzed for the obtained external loads (structural analysis. The final solution is obtained by using an iterative analysis process, which involves both aerodynamic and the structural analysis. When the solution converges, it is possible to make design modifications.

  8. A comparison of three methods for the isolation of Arcobacter spp. from retail raw poultry in Northern Ireland

    DEFF Research Database (Denmark)

    Scullion, R.; Harrington, C. S.; Madden, R. H.

    2004-01-01

    Recent evidence suggests that arcobacters, especially Arcobacter butzleri, are potential foodborne pathogens, but standardized detection methods have yet to be established. A study was undertaken to determine which of three isolation methods was the most effective for the isolation of Arcobacter...... spp. from fresh raw poultry. Method I was microaerobic and involved a membrane filtration step followed by plating onto blood agar. Method 2 was also microaerobic and involved enrichment and plating media containing a five-antibiotic cocktail. Method 3 was aerobic and was based on enrichment...... in a charcoal-based broth containing two antibiotics. Retail poultry samples (n = 50) were obtained from supermarkets in Northern Ireland; the European Community license number was recorded to ensure sample diversity. Presumptive arcobacters were identified using genus-specific and species-specific primers...

  9. Development of a direct PCR assay to detect Taenia multiceps eggs isolated from dog feces.

    Science.gov (United States)

    Wang, Ning; Wang, Yu; Ye, Qinghua; Yang, Yingdong; Wan, Jie; Guo, Cheng; Zhan, Jiafei; Gu, Xiaobin; Lai, Weimin; Xie, Yue; Peng, Xuerong; Yang, Guangyou

    2018-02-15

    Taenia multiceps is a tapeworm that leads to the death of livestock, resulting in major economic losses worldwide. The adult stage of this parasite invades the small intestine of dogs and other canids. In the present study, we developed a direct PCR assay to detect T. multiceps eggs isolated from dog feces to help curb further outbreaks. The genomic DNA was rapidly released using a lysis buffer and the PCR reaction was developed to amplify a 433-bp fragment of the T. multiceps mitochondrial gene encoding NADH dehydrogenase subunit 5 (nad5) from eggs isolated from dog feces. The procedure could be completed within 3 h, including flotation. The sensitivity of the assay was determined by detecting DNA from defined numbers of eggs, and the specificity was determined by detecting DNA from other intestinal tapeworm and roundworm species that commonly infect dogs. In addition, 14 taeniid-positive fecal samples determined by the flotation technique were collected and further evaluated by the regular PCR and our direct PCR. The results showed that the direct PCR developed herein was sensitive enough to detect the DNA from as few as 10 T. multiceps eggs and that no cross-reactions with other tapeworm and roundworm were observed, suggesting its high sensitivity and specificity for T. multiceps detection. Moreover, 14 taeniid-positive samples were screened by the regular PCR and direct PCR, with detection rates of 78.6% and 85.7%, respectively. In conclusion, the direct PCR assay developed in the present study has high sensitivity and specificity to identify T. multiceps eggs isolated from dog feces and therefore could represent an invaluable tool to identify T. multiceps outbreaks and would contribute to future clinical applications. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. Development of a novel vaccine against canine parvovirus infection with a clinical isolate of the type 2b strain.

    Science.gov (United States)

    Park, Seon Ah; Park, Seung-Yong; Song, Chang-Seon; Choi, In-Soo; Kim, Hwi Yool; Lee, Joong-Bok; Lee, Nak-Hyung

    2012-07-01

    In spite of an extensive vaccination program, parvoviral infections still pose a major threat to the health of dogs. We isolated a novel canine parvovirus (CPV) strain from a dog with enteritis. Nucleotide and amino acid sequence analysis of the isolate showed that it is a novel type 2b CPV with asparagine at the 426th position and valine at the 555th position in VP2. To develop a vaccine against CPV infection, we passaged the isolate 4 times in A72 cells. The attenuated isolate conferred complete protection against lethal homologous CPV infection in dogs such that they did not develop any clinical symptoms, and their antibody titers against CPV were significantly high at 7-11 days post infection. These results suggest that the virus isolate obtained after passaging can be developed as a novel vaccine against paroviral infection.

  11. Evaluation of Novel Wet Chemistry Separation and Purification Methods to Facilitate Automation of Astatine-211 Isolation

    International Nuclear Information System (INIS)

    Wilbur, Daniel Scott

    2016-01-01

    % extracted; There was some indication that the PEG-Merrifield resins could be saturated (perhaps with Bi) resulting in lower capture percentages, but more studies need to be done to confirm that; A target dissolution chamber, designed and built at PNNL, works well with syringe pumps so it can be used in an automated system; Preliminary semi-automated 211 At isolation studies have been conducted with full-scale target dissolution and 211 At isolation using a PEG column on the Hamilton automated system gave low overall recoveries, but HNO 3 was used (rather than HCl) for loading the 211 At and flow rates were not optimized; Results obtained using PEG columns are high enough to warrant further development on a fully automated system; Results obtained also indicate that additional studies are warranted to evaluate other types of columns for 211 At separation from bismuth, which allow use of HNO 3 /HCl mixtures for loading and NaOH for eluting 211 At. Such a column could greatly simplify the overall isolation process and make it easier to automate.

  12. Applicability of aseismic base isolation design to FBRs and state of its development

    International Nuclear Information System (INIS)

    Morishita, Masaki

    1987-01-01

    In the reactor vessels and other main equipment of FBRs, thermal stress and the stress due to earthquake load are the main dominant factors in the structural design. As to thermal stress, it is desirable to make vessels and pipings into the thin walled flexible structures, while in the aspect of aseismatic safety, it is necessary to give the required rigidity to the system. Accordingly, it is the key subject to find out the point of compromise between thermal stress-withstanding design and aseismatic design for the optimization. In the case of FBRs, particularly the rationalization of aseismatic design is necessary, and as one of the promising measures, the application of aseismic base isolation is conceivable. It is to suppress the transmission of earthquake motion to the system by designing the natural period of a structural system longer than the dominant period component of earthquake motion input. The basic concept of aseismatic base isolation design, the examples of its application in foreign countries, its research and development in Japan, the structural features of FBRs and their aseismatic base isolation design, the examples of the examination of its application to FBRs, and the problems of hereafter are described. (Kako, I.)

  13. Isolated Ficus trees deliver dual conservation and development benefits in a rural landscape.

    Science.gov (United States)

    Cottee-Jones, H Eden W; Bajpai, Omesh; Chaudhary, Lal B; Whittaker, Robert J

    2015-11-01

    Many of the world's rural populations are dependent on the local provision of economically and medicinally important plant resources. However, increasing land-use intensity is depleting these resources, reducing human welfare, and thereby constraining development. Here we investigate a low cost strategy to manage the availability of valuable plant resources, facilitated by the use of isolated Ficus trees as restoration nuclei. We surveyed the plants growing under 207 isolated trees in Assam, India, and categorized them according to their local human-uses. We found that Ficus trees were associated with double the density of important high-grade timber, firewood, human food, livestock fodder, and medicinal plants compared to non-Ficus trees. Management practices were also important in determining the density of valuable plants, with grazing pressure and land-use intensity significantly affecting densities in most categories. Community management practices that conserve isolated Ficus trees, and restrict livestock grazing and high-intensity land-use in their vicinity, can promote plant growth and the provision of important local resources.

  14. Reactor kinetics methods development. Final report

    International Nuclear Information System (INIS)

    Hansen, K.F.; Henry, A.F.

    1978-01-01

    This report is a qualitative summary of research conducted at MIT from 1967 to 1977 in the area of reactor kinetics methods. The objectives of the research were to find methods of integration of various mathematical models of nuclear reactor transients. From the beginning the work was aimed at numerical integration methods. Specific areas of research, discussed in more detail following, included: integration of multigroup diffusion theory models by finite difference and finite element methods; response matrix and nodal methods; coarse-mesh homogenization; and special treatment of boundary conditions

  15. [Isolation methods and diversity of culturable fecal actinobacteria associated with Panthera tigris tigris in Yunnan Safari Park].

    Science.gov (United States)

    Cao, Yanru; Jiang, Yi; Li, Youlong; Chen, Xiu; Jin, Rongxian; He, Wenxiang

    2012-07-04

    We studied the isolation methods and diversity of culturable fecal actinobacteria associated with Panthera tigris tigris by using culture-dependent approaches. Fresh fecal samples of healthy Panthera tigris tigris were collected from Yunnan Safari Park. Pretreatment of the samples, isolation media and inhibitors were tested for actinobacteria isolation. 16S rRNA genes of actinobacteria were sequenced and subjected to phylogenetic analysis. The abundance of culturable actinobacteria was 1.10 x 10(8) cfu/g colony forming units (CFU) per gram of feces (wet weight). We obtained 110 purified cultural actinobacterium strains. The analysis based on 16S rRNA gene sequences showed that these strains were distributed in 10 different families and 12 genera of actinobacteria at least, and most of them were non-filamentous, such as Arthrobacter, Dietzia, Kocuria, Corynebacterium and Microbacterium. Streptomyces was the mainly classical filamentous actinobacteria, and up to 64% of total. Drying and heating up the fecal samples can greatly increase the rate of the actinobacteria. Many kinds of inhibitors and chemical defined media are suitable for isolation of fecal actinobacteria. The culturable actinobacteria are abundant in Panthera tigris tigris feces. Our study found an effective method to isolate animals' fecal actinobacteria and it's useful for studying and exploiting animals' fecal actinobacteria.

  16. Isolation of αL I domain mutants mediating firm cell adhesion using a novel flow-based sorting method.

    Science.gov (United States)

    Pepper, Lauren R; Parthasarathy, Ranganath; Robbins, Gregory P; Dang, Nicholas N; Hammer, Daniel A; Boder, Eric T

    2013-08-01

    The inserted (I) domain of αLβ2 integrin (LFA-1) contains the entire binding site of the molecule. It mediates both rolling and firm adhesion of leukocytes at sites of inflammation depending on the activation state of the integrin. The affinity change of the entire integrin can be mimicked by the I domain alone through mutations that affect the conformation of the molecule. High-affinity mutants of the I domain have been discovered previously using both rational design and directed evolution. We have found that binding affinity fails to dictate the behavior of I domain adhesion under shear flow. In order to better understand I domain adhesion, we have developed a novel panning method to separate yeast expressing a library of I domain variants on the surface by adhesion under flow. Using conditions analogous to those experienced by cells interacting with the post-capillary vascular endothelium, we have identified mutations supporting firm adhesion that are not found using typical directed evolution techniques that select for tight binding to soluble ligands. Mutants isolated using this method do not cluster with those found by sorting with soluble ligand. Furthermore, these mutants mediate shear-driven cell rolling dynamics decorrelated from binding affinity, as previously observed for I domains bearing engineered disulfide bridges to stabilize activated conformational states. Characterization of these mutants supports a greater understanding of the structure-function relationship of the αL I domain, and of the relationship between applied force and bioadhesion in a broader context.

  17. Early social isolation impairs development, mate choice and grouping behaviour of predatory mites.

    Science.gov (United States)

    Schausberger, Peter; Gratzer, Marian; Strodl, Markus A

    2017-05-01

    The social environment early in life is a key determinant of developmental, physiological and behavioural trajectories across vertebrate and invertebrate animals. One crucial variable is the presence/absence of conspecifics. For animals usually reared in groups, social isolation after birth or hatching can be a highly stressful circumstance, with potentially long-lasting consequences. Here, we assessed the effects of social deprivation (isolation) early in life, that is, absence of conspecifics, versus social enrichment, that is, presence of conspecifics, on developmental time, body size at maturity, mating behaviour and group-living in the plant-inhabiting predatory mite Phytoseiulus persimilis . Socially deprived protonymphs developed more slowly and were less socially competent in grouping behaviour than socially enriched protonymphs. Compromised social competence in grouping behaviour was evident in decreased activity, fewer mutual encounters and larger interindividual distances, all of which may entail severe fitness costs. In female choice/male competition, socially deprived males mated earlier than socially enriched males; in male choice/female competition, socially deprived females were more likely to mate than socially enriched females. In neither mate choice situation did mating duration or body size at maturity differ between socially deprived and enriched mating opponents. Social isolation-induced shifts in mating behaviour may be interpreted as increased attractiveness or competitiveness or, more likely, as hastiness and reduced ability to assess mate quality. Overall, many of the social isolation-induced behavioural changes in P. persimilis are analogous to those observed in other animals such as cockroaches, fruit flies, fishes or rodents. We argue that, due to their profound and persistent effects, early social deprivation or enrichment may be important determinants in shaping animal personalities.

  18. Comparison of some storage and isolation methods to recover bluetongue virus from bovine blood.

    OpenAIRE

    Thomas, F C

    1984-01-01

    Bovine blood containing bluetongue virus was stored as whole blood with EDTA (4 degrees C), as washed cellular components (4 degrees C), and as washed cellular components with 10% DMSO (-70 degrees C). Periodic isolation attempts were made over a period of 330 days in four cell lines and embryonating chicken eggs (intravenous inoculation). Bluetongue virus was successfully isolated in all systems from most samples throughout the test period. There appeared to be more variation amongst days of...

  19. Characterization of single spore isolates of Agaricus bisporus (Lange) Imbach using conventional and molecular methods.

    Science.gov (United States)

    Sharma, Manju; Suman, B C; Gupta, Dharmesh

    2014-10-01

    Strains A-15, S11, S-140, and U3 of Agaricus bisporus (Lange) Imbach, were used as parent strains for raising single spore homokaryotic isolates. Out of total 1,642 single spore isolates, only 36 single spore isolates were homokaryons and exhibited slow mycelial growth rate (≤2.0 mm/day) and appressed colony morphology. All these SSIs failed to produce pinheads in Petri plates even after 65 days of incubation, whereas the strandy slow growing SSIs along with parent strains were able to form the fructification in petriplates after 30 days. Out of 24, six ISSR primers, exhibited scorable bands. In the ISSR fingerprints, single spore isolates, homokaryons, lacked amplification products at multiple loci; they grow slowly and all of them had appressed types of colony morphology. The study revealed losses of ISSR polymorphic patterns in non-fertile homokaryotic single spore isolates compared to the parental control or fertile heterokaryotic single spore isolates.

  20. Development of radiolabeling method for natural juvenile hormones

    International Nuclear Information System (INIS)

    Okuda, Takashi; Shiotsuki, Takahiro; Kotaki, Toyomi

    2000-01-01

    This study aimed to develop a new assay method for accurate determination of juvenile hormone (JH) using radioisotope. A new measuring method for JH was designed based on the principle of molecular competition. Namely, JH-binding protein in the insect was used to form a selective binding to JH. At first, corpus allatum was cultured in a medium containing 3 H or 14 C epoxyfarnesyldiazoacetate (EFDA), which is a photoaffinity labeling reagent and JH binding protein (JHBP) was purified using 3 H or 14 C labeled EFDA. Since several kinds of JH homologues different in the molecular structure have been known, it was needed to establish each labeling method appropriate to those homologues. Then, an assay method for micro-quantitative measurement of JH was established utilizing the competition between labeled natural JH and JHBP. Thus, the authors succeeded in the overexpression of the blood JHBA of kind of tabaco moth, H.virescens and also in cloning and overexpression of JHBP in the silk worm. It was confirmed that these JHBPs specifically bind to 3 H labeled JH3, leading to stable supply of blood JHBP. Thus, accurate assay for JH concentration became possible by the use of the labeled JH with natural configuration and the blood JHBP. In the course of this study, several findings were obtained as follows: The JHBP of a sting bug was different from the previously reported ones. The regulation of JH synthesis in the corpus allatum was closely related to the control of diapause in several insects. The JHBP isolated from the cytosol of silk gland was a new protein in respect of amino acid sequence. (M.N.)

  1. Development of radiolabeling method for natural juvenile hormones

    Energy Technology Data Exchange (ETDEWEB)

    Okuda, Takashi; Shiotsuki, Takahiro; Kotaki, Toyomi [National Inst. of Sericultural and Entomological Science, Tsukuba, Ibaraki (Japan)

    2000-02-01

    This study aimed to develop a new assay method for accurate determination of juvenile hormone (JH) using radioisotope. A new measuring method for JH was designed based on the principle of molecular competition. Namely, JH-binding protein in the insect was used to form a selective binding to JH. At first, corpus allatum was cultured in a medium containing {sup 3}H or {sup 14}C epoxyfarnesyldiazoacetate (EFDA), which is a photoaffinity labeling reagent and JH binding protein (JHBP) was purified using {sup 3}H or {sup 14}C labeled EFDA. Since several kinds of JH homologues different in the molecular structure have been known, it was needed to establish each labeling method appropriate to those homologues. Then, an assay method for micro-quantitative measurement of JH was established utilizing the competition between labeled natural JH and JHBP. Thus, the authors succeeded in the overexpression of the blood JHBA of kind of tabaco moth, H.virescens and also in cloning and overexpression of JHBP in the silk worm. It was confirmed that these JHBPs specifically bind to {sup 3}H labeled JH3, leading to stable supply of blood JHBP. Thus, accurate assay for JH concentration became possible by the use of the labeled JH with natural configuration and the blood JHBP. In the course of this study, several findings were obtained as follows: The JHBP of a sting bug was different from the previously reported ones. The regulation of JH synthesis in the corpus allatum was closely related to the control of diapause in several insects. The JHBP isolated from the cytosol of silk gland was a new protein in respect of amino acid sequence. (M.N.)

  2. Comparison of biochemical and molecular methods for the identification of bacterial isolates associated with failed loggerhead sea turtle eggs.

    Science.gov (United States)

    Awong-Taylor, J; Craven, K S; Griffiths, L; Bass, C; Muscarella, M

    2008-05-01

    Comparison of biochemical vs molecular methods for identification of microbial populations associated with failed loggerhead turtle eggs. Two biochemical (API and Microgen) and one molecular methods (16s rRNA analysis) were compared in the areas of cost, identification, corroboration of data with other methods, ease of use, resources and software. The molecular method was costly and identified only 66% of the isolates tested compared with 74% for API. A 74% discrepancy in identifications occurred between API and 16s rRNA analysis. The two biochemical methods were comparable in cost, but Microgen was easier to use and yielded the lowest discrepancy among identifications (29%) when compared with both API 20 enteric (API 20E) and API 20 nonenteric (API 20NE) combined. A comparison of API 20E and API 20NE indicated an 83% discrepancy between the two methods. The Microgen identification system appears to be better suited than API or 16s rRNA analysis for identification of environmental isolates associated with failed loggerhead eggs. Most identification methods are not intended for use with environmental isolates. A comparison of identification systems would provide better options for identifying environmental bacteria for ecological studies.

  3. An efficient method for isolating antibody fragments against small peptides by antibody phage display

    DEFF Research Database (Denmark)

    Duan, Zhi; Siegumfeldt, Henrik

    2010-01-01

    We generated monoclonal scFv (single chain variable fragment) antibodies from an antibody phage display library towards three small synthetic peptides derived from the sequence of s1-casein. Key difficulties for selection of scFv-phages against small peptides were addressed. Small peptides do....... The scFvs were sequenced and characterized, and specificity was characterized by ELISA. The methods developed in this study are universally applicable for antibody phage display to efficiently produce antibody fragments against small peptides....

  4. A peer review of the Hanford Site Permanent Isolation Surface Barrier Development Program

    International Nuclear Information System (INIS)

    Wing, N.R.

    1992-09-01

    A panel of technical experts was organized to peer review the Hanford Site Permanent Isolation Surface Barrier Development Program (BDP) and to provide a specific review of a preconceptual prototype barrier design initiated during fiscal year (FY) 1990. The technical peer review of the BDP and the prototype is being conducted in three phases, two of which have been completed. This document presents the peer review panel's findings on the first two phases of the peer review process. Biointrusion and water intrusion control are discussed, along with design life, vegetation, and climate impact

  5. Microsatellite marker isolation and development for the giant Pacific Octopus (Enteroctopus dofleini)

    Science.gov (United States)

    Toussaint, Rebecca K.; Sage, G. Kevin; Talbot, Sandra L.; Scheel, David

    2012-01-01

    We isolated and developed 18 novel microsatellite markers for the giant Pacific octopus (Enteroctopus dofleini) and examined them for 31 individuals from Prince William Sound (PWS), Alaska. These loci displayed moderate levels of allelic diversity (averaging 11 alleles per locus) and heterozygosity (averaging 65%). Seven loci deviated from Hardy–Weinberg Equilibrium (HWE) due to heterozygote deficiency for the PWS population, although deviations were not observed for all these loci in other populations, suggesting the PWS population is not in mutation-drift equilibrium. These novel microsatellite loci yielded sufficient genetic diversity for potential use in population genetics, individual identification, and parentage studies.

  6. Minutes of 'Technical Committee for Geological Isolation Research and Development (1st time)'

    International Nuclear Information System (INIS)

    Sasaki, Yasuo; Torata, Shinichiro

    2007-03-01

    In order to plan and conduct a program of research and development (R and D) for geological disposal of high-level radioactive waste in JAEA-Tokai, JAEA decided to receive technical comments and advices from the Technical Committee for geological isolation R and D. This committee, which is constituted of specialists who do not belong to JAEA, has evaluated the program, taking into account of the status of geological disposal technology in Japan as well as foreign countries. This report compiles the minutes of the first meeting held in July 2006. (author)

  7. An adhesion-based method for plasma membrane isolation: evaluating cholesterol extraction from cells and their membranes.

    Science.gov (United States)

    Bezrukov, Ludmila; Blank, Paul S; Polozov, Ivan V; Zimmerberg, Joshua

    2009-11-15

    A method to isolate large quantities of directly accessible plasma membrane from attached cells is presented. The method is based on the adhesion of cells to an adsorbed layer of polylysine on glass plates, followed by hypotonic lysis with ice-cold distilled water and subsequent washing steps. Optimal conditions for coating glass plates and time for cell attachment were established. No additional chemical or mechanical treatments were used. Contamination of the isolated plasma membrane by cell organelles was less than 5%. The method uses inexpensive, commercially available polylysine and reusable glass plates. Plasma membrane preparations can be made in 15 min. Using this method, we determined that methyl-beta-cyclodextrin differentially extracts cholesterol from fibroblast cells and their plasma membranes and that these differences are temperature dependent. Determination of the cholesterol/phospholipid ratio from intact cells does not reflect methyl-beta-cyclodextrin plasma membrane extraction properties.

  8. Toxigenic potentiality of Aspergillus flavus and Aspergillus parasiticus strains isolated from black pepper assessed by an LC-MS/MS based multi-mycotoxin method.

    Science.gov (United States)

    Yogendrarajah, Pratheeba; Devlieghere, Frank; Njumbe Ediage, Emmanuel; Jacxsens, Liesbeth; De Meulenaer, Bruno; De Saeger, Sarah

    2015-12-01

    A liquid chromatography triple quadrupole tandem mass spectrometry method was developed and validated to determine mycotoxins, produced by fungal isolates grown on malt extract agar (MEA). All twenty metabolites produced by different fungal species were extracted using acetonitrile/1% formic acid. The developed method was applied to assess the toxigenic potentiality of Aspergillus flavus (n = 11) and Aspergillus parasiticus (n = 6) strains isolated from black peppers (Piper nigrum L.) following their growth at 22, 30 and 37 °C. Highest mean radial colony growth rates were observed at 30 °C for A. flavus (5.21 ± 0.68 mm/day) and A. parasiticus (4.97 ± 0.33 mm/day). All of the A. flavus isolates produced aflatoxin B1 and O-methyl sterigmatocystin (OMST) while 91% produced aflatoxin B2 (AFB2) and 82% of them produced sterigmatocystin (STERIG) at 30 °C. Except one, all the A. parasiticus isolates produced all the four aflatoxins, STERIG and OMST at 30 °C. Remarkably high AFB1 was produced by some A. flavus isolates at 22 °C (max 16-40 mg/kg). Production of mycotoxins followed a different trend than that of growth rate of both species. Notable correlations were found between different secondary metabolites of both species; R(2) 0.87 between AFB1 and AFB2 production. Occurrence of OMST could be used as a predictor for AFB1 production. Copyright © 2015 Elsevier Ltd. All rights reserved.

  9. Intercomparison of analysis methods for seismically isolated nuclear structures. Papers and working materials presented at the 3. research coordination meeting

    International Nuclear Information System (INIS)

    1998-01-01

    The Coordinated research program on Intercomparison of analysis methods for seismically isolated nuclear structures involved participants from Italy, Japan, Republic of Korea, Russia, United Kingdom, USA, EC. The purpose of the meeting was to review the progress on the finite element prediction of the force-deformation behaviour of seismic isolators and to discuss the first set of analytical results for the prediction of the response of base-oscillated structures to earthquake inputs. The intercomparison of predictions of bearing behaviour has identified important unexpected issues requiring deeper investigation

  10. HPTLC and reverse phase HPLC methods for the simultaneous quantification and in vitro screening of antioxidant potential of isolated sesquiterpenoids from the rhizomes of Cyperus rotundus.

    Science.gov (United States)

    Priya Rani, M; Padmakumari, K P

    2012-09-01

    Three sesquiterpenoids solavetivone, aristolone and nootkatone were isolated from the acetone extract of Cyperus rotundus by silica gel column chromatography and identified by spectral studies. Solavetivone has been isolated for the first time from the species. Simple, sensitive and selective HPTLC and HPLC methods with ultraviolet detection (245 nm) were developed and validated for the simultaneous quantification. HPTLC method was validated in terms of their linearity, LOD, LOQ, precision, accuracy and compared with RP-HPLC-UV method. Among the three sesquiterpenoids isolated, nootkatone possessed the highest radical scavenging potential (IC(50) 4.81 μg/ml) followed by aristolone (IC(50) 5.28 μg/ml) and solavetivone (IC(50) 6.82 μg/ml) by DPPH radical scavenging assay. Total antioxidant activity against phosphomolybdenum reagent was also studied. The methods described in this paper were able to identify and quantify sesquiterpenoids from the complex mixtures of phytochemicals and could be extended to the marker based standardization of polyherbal formulations containing C. rotundus. Copyright © 2012 Elsevier B.V. All rights reserved.

  11. Development on Vulnerability Assessment Methods of PPS

    Institute of Scientific and Technical Information of China (English)

    MIAO; Qiang; ZHANG; Wen-liang; BU; Li-xin; YIN; Hong-he; LI; Xin-jun; FANG; Xin

    2013-01-01

    Through investigating information from domestic and abroad,joint the domestic assessment experience,we present a set of physical protection system(PPS)vulnerability assessment methods for on-operating nuclear power plants and for on-designing nuclear facilities.The methods will help to strengthen and upgrade the security measures of the nuclear facilities,improve the effectiveness and

  12. Development and Application of Kinetic Spectrophotometric Method ...

    African Journals Online (AJOL)

    ISSN: 1596-5996 (print); 1596-9827 (electronic) ... Methods: The method is based on oxidation reaction of MNZ by hydrogen peroxide ... optimum operating conditions for reagent concentrations and temperature were ... 1-yl) ethanol] is an amebicide, antiprotozoal and .... The dependence of reaction rate on concentration of.

  13. Scenario development for the Waste Isolation Pilot Plant: Building confidence in the assessment

    International Nuclear Information System (INIS)

    Galson, D.A.; Swift, P.N.

    1994-01-01

    Scenario developments is part of the iterative performance assessment (PA) process for the Waste Isolation Pilot Plant (WIPP). Scenario development for the WIPP has been the subject of intense external review, and is certain to be the subject of continued scrutiny as the project proceeds toward regulatory compliance. The principal means of increasing confidence is this aspect of the PA will be through the use of a systematic and thorough procedure toward developing the scenarios and conceptual models on which the assessment is to be based. Early and ongoing interaction with project reviewers can assist with confidence building. Quality of argument and clarity of presentation in PA will be of key concern. Appropriate tools are required for documenting and tracking assumptions, through a single assessment phase, and between iterative assessment phases. Risks associated with future human actions are of particular concern to the WIPP project, and international consensus on the principles for incorporation of future human actions in assessments would be valuable

  14. Scenario development for the Waste Isolation Pilot Plant: Building confidence in the assessment

    International Nuclear Information System (INIS)

    Galson, D.A.; Swift, P.N.

    1994-07-01

    Scenario development is part of the iterative performance assessment (PA) process for the Waste Isolation Pilot Plant (WIPP). Scenario development for the WIPP has been the subject of intense external review and is certain to be the subject of continued scrutiny as the project proceeds toward regulatory compliance. The principal means of increasing confidence in this aspect of the PA will be through the use of the systematic and thorough procedure toward developing the scenarios and conceptual models on which the assessment is to be based. Early and ongoing interaction with project reviewers can assist with confidence building. Quality of argument and clarity of presentation in PA will be of key concern. Appropriate tools are required for documenting and tracking assumptions, through a single assessment phase, and between iterative assessment phases. Risks associated with future human actions are of particular concern to the WIPP project, and international consensus on the principles for incorporation of future human actions in assessments would be valuable

  15. DEVELOPMENT OF A MOLECULAR METHOD TO IDENTIFY HEPATITIS E VIRUS

    Science.gov (United States)

    Hepatitis E virus (HEV) is a waterborne emerging pathogen that causes significant illness in the developing world. Thus far, an HEV outbreak has not been reported in the U.S., although a swine variant of the virus is common in Midwestern hogs. Because viruses isolated from two ...

  16. Comparison of blood RNA isolation methods from samples stabilized in Tempus tubes and stored at a large human biobank.

    Science.gov (United States)

    Aarem, Jeanette; Brunborg, Gunnar; Aas, Kaja K; Harbak, Kari; Taipale, Miia M; Magnus, Per; Knudsen, Gun Peggy; Duale, Nur

    2016-09-01

    More than 50,000 adult and cord blood samples were collected in Tempus tubes and stored at the Norwegian Institute of Public Health Biobank for future use. In this study, we systematically evaluated and compared five blood-RNA isolation protocols: three blood-RNA isolation protocols optimized for simultaneous isolation of all blood-RNA species (MagMAX RNA Isolation Kit, both manual and semi-automated protocols; and Norgen Preserved Blood RNA kit I); and two protocols optimized for large RNAs only (Tempus Spin RNA, and Tempus 6-port isolation kit). We estimated the following parameters: RNA quality, RNA yield, processing time, cost per sample, and RNA transcript stability of six selected mRNAs and 13 miRNAs using real-time qPCR. Whole blood samples from adults (n = 59 tubes) and umbilical cord blood (n = 18 tubes) samples collected in Tempus tubes were analyzed. High-quality blood-RNAs with average RIN-values above seven were extracted using all five RNA isolation protocols. The transcript levels of the six selected genes showed minimal variation between the five protocols. Unexplained differences within the transcript levels of the 13 miRNA were observed; however, the 13 miRNAs had similar expression direction and they were within the same order of magnitude. Some differences in the RNA processing time and cost were noted. Sufficient amounts of high-quality RNA were obtained using all five protocols, and the Tempus blood RNA system therefore seems not to be dependent on one specific RNA isolation method.

  17. Shigella isolates from the global enteric multicenter study inform vaccine development.

    Science.gov (United States)

    Livio, Sofie; Strockbine, Nancy A; Panchalingam, Sandra; Tennant, Sharon M; Barry, Eileen M; Marohn, Mark E; Antonio, Martin; Hossain, Anowar; Mandomando, Inacio; Ochieng, John B; Oundo, Joseph O; Qureshi, Shahida; Ramamurthy, Thandavarayan; Tamboura, Boubou; Adegbola, Richard A; Hossain, Mohammed Jahangir; Saha, Debasish; Sen, Sunil; Faruque, Abu Syed Golam; Alonso, Pedro L; Breiman, Robert F; Zaidi, Anita K M; Sur, Dipika; Sow, Samba O; Berkeley, Lynette Y; O'Reilly, Ciara E; Mintz, Eric D; Biswas, Kousick; Cohen, Dani; Farag, Tamer H; Nasrin, Dilruba; Wu, Yukun; Blackwelder, William C; Kotloff, Karen L; Nataro, James P; Levine, Myron M

    2014-10-01

    Shigella, a major diarrheal disease pathogen worldwide, is the target of vaccine development. The Global Enteric Multicenter Study (GEMS) investigated burden and etiology of moderate-to-severe diarrheal disease in children aged Shigella was 1 of the 4 most common pathogens across sites and age strata. GEMS Shigella serotypes are reviewed to guide vaccine development. Subjects' stool specimens/rectal swabs were transported to site laboratories in transport media and plated onto xylose lysine desoxycholate and MacConkey agar. Suspect Shigella colonies were identified by biochemical tests and agglutination with antisera. Shigella isolates were shipped to the GEMS Reference Laboratory (Baltimore, MD) for confirmation and serotyping of S. flexneri; one-third of isolates were sent to the Centers for Disease Control and Prevention for quality control. Shigella dysenteriae and S. boydii accounted for 5.0% and 5.4%, respectively, of 1130 Shigella case isolates; S. flexneri comprised 65.9% and S. sonnei 23.7%. Five serotypes/subserotypes comprised 89.4% of S. flexneri, including S. flexneri 2a, S. flexneri 6, S. flexneri 3a, S. flexneri 2b, and S. flexneri 1b. A broad-spectrum Shigella vaccine must protect against S. sonnei and 15 S. flexneri serotypes/subserotypes. A quadrivalent vaccine with O antigens from S. sonnei, S. flexneri 2a, S. flexneri 3a, and S. flexneri 6 can provide broad direct coverage against these most common serotypes and indirect coverage against all but 1 (rare) remaining subserotype through shared S. flexneri group antigens. © The Author 2014. Published by Oxford University Press on behalf of the Infectious Diseases Society of America.

  18. Production, Isolation and Radiolabeling Methods for 211AT- Labeling of Biomolecules

    International Nuclear Information System (INIS)

    Wilbur, D.S.; Hamlin, D.K.; Chyan, M.

    2009-01-01

    Targeted alpha therapy with 211 At-labeled compounds holds great promise for treatment of cancer, particularly compartmentalized cancer (e.g. ovarian cancer), minimal residual cancer after surgery and metastatic disease. Unfortunately, 211 At has limited availability and, due to its unique nature, has the potential to be readily dissociated from the cancer-targeting agents used in vivo. Finding methods to circumvent these two problems has occupied a large amount of our efforts over the past few years. 211 At is produced at the University of Washington on a Scanditronix MC-50 using a 28 MeV alpha beam. Our initial preclinical studies were conducted using a small target assembly with irradiations of a 10 □ A alpha beam, but our desire to ultimately conduct clinical studies led to the design and installation of a new target assembly that had much larger irradiation surface and would withstand beam energies of 50 □A or more. Prior to this upgrade, 211 At was efficiently isolated (60-80%) from the irradiated aluminum-backed bismuth targets by dry distillation at 650 o C. However, the dry distillation method gave low recovery yields (e.g. 10-40%) when the much larger new targets were used. After some attempts to improve the distillation yields, we have more recently conducted a wet chemistry approach to the 211 At isolation. While this method still needs to be optimized, it has provided good recovery (60-90%) of the 211 At. Our radiolabeling methods have undergone a similar transition in the past few years. Until recently our 211 At studies were limited to the use of intact monoclonal antibodies (MAb) labeled using conjugates containing aryl-astatine derivatives due to the deastatination of more rapidly metabolized targeting biomolecules. This limitation made it all but impossible to label important biomolecules such as MAb fragments, engineered proteins, peptides and small molecules. This critical shortcoming of labeling methods for 211 At led to our investigating

  19. Detection of Chloramphenicol Resistance Genes (cat in Clinical Isolates of Pseudomonas aeruginosa with Polymerase Chain Reaction Method

    Directory of Open Access Journals (Sweden)

    Tiana Milanda

    2014-12-01

    Full Text Available Pseudomonas aeruginosa is an opportunistic Gram negative bacteria, which may cause infection in eyes, ears, skin, bones, central nervous system, gastrointestinal tract, circulatory system, heart, respiratory system, and urinary tract. Recently, chloramphenicol is no longer used as the main option of the therapy due of its resistance case. The aim of this research was to detect the presence of gene which is responsible to chloramphenicol resistance in clinical isolates of P.aeruginosa. These bacteria isolated from pus of external otitis patients in Hasan Sadikin Hospital in Bandung City. Polymerase Chain Reaction (PCR method (colony-PCR and DNA-PCR were performed to detect this resistance gene. Electropherogram from PCR products showed that the chloramphenicol resistance in clinical isolates of P. aeruginosa was caused by cat gene (317 bp. Based on this research, cat gene may be used to detect the chloramphenicol resistance in patients with external ostitis.

  20. A simple and rapid method for isolation of high quality genomic DNA from fruit trees and conifers using PVP.

    Science.gov (United States)

    Kim, C S; Lee, C H; Shin, J S; Chung, Y S; Hyung, N I

    1997-03-01

    Because DNA degradation is mediated by secondary plant products such as phenolic terpenoids, the isolation of high quality DNA from plants containing a high content of polyphenolics has been a difficult problem. We demonstrate an easy extraction process by modifying several existing ones. Using this process we have found it possible to isolate DNAs from four fruit trees, grape (Vitis spp.), apple (Malus spp.), pear (Pyrus spp.) and persimmon (Diospyros spp.) and four species of conifer, Pinus densiflora, Pinus koraiensis,Taxus cuspidata and Juniperus chinensis within a few hours. Compared with the existing method, we have isolated high quality intact DNAs (260/280 = 1.8-2.0) routinely yielding 250-500 ng/microl (total 7.5-15 microg DNA from four to five tissue discs).

  1. Molecular profiles of Venezuelan isolates of Trypanosoma sp. by random amplified polymorphic DNA method.

    Science.gov (United States)

    Perrone, T M; Gonzatti, M I; Villamizar, G; Escalante, A; Aso, P M

    2009-05-12

    Nine Trypanosoma sp. Venezuelan isolates, initially presumed to be T. evansi, were collected from three different hosts, capybara (Apure state), horse (Apure state) and donkey (Guarico state) and compared by the random amplification polymorphic DNA technique (RAPD). Thirty-one to 46 reproducible fragments were obtained with 12 of the 40 primers that were used. Most of the primers detected molecular profiles with few polymorphisms between the seven horse, capybara and donkey isolates. Quantitative analyses of the RAPD profiles of these isolates revealed a high degree of genetic conservation with similarity coefficients between 85.7% and 98.5%. Ten of the primers generated polymorphic RAPD profiles with two of the three Trypanosoma sp. horse isolates, namely TeAp-N/D1 and TeGu-N/D1. The similarity coefficient between these two isolates and the rest, ranged from 57.9% to 68.4% and the corresponding dendrogram clustered TeAp-N/D1 and Te Gu-N/D1 in a genetically distinct group.

  2. The isolation of nucleic acids from fixed, paraffin-embedded tissues-which methods are useful when?

    DEFF Research Database (Denmark)

    Gilbert, M Thomas P; Haselkorn, Tamara; Bunce, Michael

    2007-01-01

    . Cross-linking not only complicates isolation of nucleic acid but also introduces polymerase "blocks" during PCR. A wide variety of methods exists for the recovery of DNA and RNA from archival tissues, and although a number of previous studies have qualitatively compared the relative merits....... These include methods of pre-treating the samples prior to extraction, extraction and nucleic acid purification methods themselves, and a post-extraction enzymatic repair technique. We find that although many of the published methods have distinct positive effects on some characteristics of the nucleic acids...

  3. Development of Ultraviolet Spectrophotometric Method for Analysis ...

    African Journals Online (AJOL)

    HP

    Method for Analysis of Lornoxicam in Solid Dosage. Forms. Sunit Kumar Sahoo ... testing. Mean recovery was 100.82 % for tablets. Low values of % RSD indicate .... Saharty E, Refaat YS, Khateeb ME. Stability-. Indicating. Spectrophotometric.

  4. Developing methods of controlling quality costs

    OpenAIRE

    Gorbunova A. V.; Maximova O. N.; Ekova V. A.

    2017-01-01

    The article examines issues of managing quality costs, problems of applying economic methods of quality control, implementation of progressive methods of quality costs management in enterprises with the view of improving the efficiency of their evaluation and analysis. With the aim of increasing the effectiveness of the cost management mechanism, authors introduce controlling as a tool of deviation analysis from the standpoint of the process approach. A list of processes and corresponding eva...

  5. Bayesian methods for proteomic biomarker development

    Directory of Open Access Journals (Sweden)

    Belinda Hernández

    2015-12-01

    In this review we provide an introduction to Bayesian inference and demonstrate some of the advantages of using a Bayesian framework. We summarize how Bayesian methods have been used previously in proteomics and other areas of bioinformatics. Finally, we describe some popular and emerging Bayesian models from the statistical literature and provide a worked tutorial including code snippets to show how these methods may be applied for the evaluation of proteomic biomarkers.

  6. The Isolation of Pure Populations of Neurons by Laser Capture Microdissection: Methods and Application in Neuroscience.

    Science.gov (United States)

    Morris, Renée; Mehta, Prachi

    2018-01-01

    In mammals, the central nervous system (CNS) is constituted of various cellular elements, posing a challenge to isolating specific cell types to investigate their expression profile. As a result, tissue homogenization is not amenable to analyses of motor neurons profiling as these represent less than 10% of the total spinal cord cell population. One way to tackle the problem of tissue heterogeneity and obtain meaningful genomic, proteomic, and transcriptomic profiling is to use laser capture microdissection technology (LCM). In this chapter, we describe protocols for the capture of isolated populations of motor neurons from spinal cord tissue sections and for downstream transcriptomic analysis of motor neurons with RT-PCR. We have also included a protocol for the immunological confirmation that the captured neurons are indeed motor neurons. Although focused on spinal cord motor neurons, these protocols can be easily optimized for the isolation of any CNS neurons.

  7. Phenotypic methods for detection of various β-lactamases in Gram-negative clinical isolates: Need of the hour

    Directory of Open Access Journals (Sweden)

    Neena V Nagdeo

    2012-01-01

    Full Text Available Background: Many clinical laboratories have problems detecting various β-lactamases. Confusion exists about the importance of these resistance mechanisms, optimal test methods, and appropriate reporting conventions. It is more imperative to use various phenotypic methods for detection of various β-lactamases in routine microbiology laboratory on day-to-day basis to prevent antimicrobial resistance by evidence-based judicious use of antimicrobials. Aims: In view of the multidrug-resistant organisms being reported world over, we planned a cross-sectional prospective analytical study to determine resistance mechanism by various β-lactamases in Gram-negative clinical isolates using various phenotypic methods. Materials and Methods: All nonrepeat, nonenteric clinical isolates of Gram-negative bacilli, resistant to at least two third-generation cephalosporins, were first screened by Novel disc placement method, and isolates showing multiple mechanisms of resistance and reduced zone of inhibition for imipenem were further confirmed for AmpC and metallo β-lactamases. Statistical Analysis: All the data was managed and analyzed in Microsoft Excel. Results: Out of 807 isolates tested, as many as 795 (98.51% revealed the presence of extended-spectrum β-lactamases (ESBLs. Only 10 isolates of Escherichia coli and 2 of Klebsiella pneumoniae did not show production of ESBL. A total of 450 (55.76% isolates produced single enzyme,while 345 (42.75% strains revealed multiple enzyme production simultaneously. Only ESBL production was seen in 315 (39.03% strains, only AmpC in 75 (9.29% and only MBL in 60 (7.44% strains, while ESBL and AmpC together were seen in 219 (27.14% and AmpC plus MBL in 92 (11.40% strains. However, ESBL plus MBL were never observed together. All three enzymes were simultaneously detected in 34 (4.21% strains. Conclusion: This innovative method of disc placement makes it easy, affordable, and reliable method for routine use by basic

  8. Comparative study of different methods used to isolate Trypanosoma cruz i for defibrinated blood of irradiated mice

    International Nuclear Information System (INIS)

    Fontes, G.; Romanha, A.J.; Brener, Z.

    1991-01-01

    Different methods are being used for the isolation and purification of Trypanosoma cruzi blood forms from infected vertebrate hosts. In this study, we compare four of these methods (differential centrifugation, Ficoll-Hypaque, Histopaque 1077 and metrizamide) in terms of parasite recovery rates, contamination with cells, duration of the process and role of host irradiation. Male albino Swiss mice irradiated in a Gamma Cell 220(500 rads) were inoculated with C L and V-10 T.cruzi strains and bled at the peak of parasitemia. Infected defibrinated blood was then used for the isolation. Although all methods permitted the recovery of viable trypomastigotes, the best results were obtained with Ficoll-Hypaque and Histopaque 1077. Recovery rates ranged between 71% to 88% and parasite-enriched preparations were obtained in approximately 75 min. Irradiation and blood defibrination drastically reduced platelet and leukocyte contamination of the preparations. (author)

  9. TX-RX isolation method based on polarization diversity, spatial diversity and TX beamforming

    DEFF Research Database (Denmark)

    Foroozanfard, Ehsan; Carvalho, Elisabeth De; Pedersen, Gert F.

    2016-01-01

    In this paper, the feasibility of an antenna isolation technique based on null-steer beamforming, polarization diversity and spatial diversity is investigated. The proposed system consists of six patch antennas which are fed by a feeding network to obtain a null-steer beamformer. To achieve spatial...... diversity, antenna elements are located on two layers, facing in a different direction. Moreover, the antenna elements in two layers use different polarization. The measured results of the antenna system present a high TX-RX isolation in the order of 70 dB which shows the feasibility of such a system...

  10. Thermally-isolated silicon-based integrated circuits and related methods

    Science.gov (United States)

    Wojciechowski, Kenneth; Olsson, Roy H.; Clews, Peggy J.; Bauer, Todd

    2017-05-09

    Thermally isolated devices may be formed by performing a series of etches on a silicon-based substrate. As a result of the series of etches, silicon material may be removed from underneath a region of an integrated circuit (IC). The removal of the silicon material from underneath the IC forms a gap between remaining substrate and the integrated circuit, though the integrated circuit remains connected to the substrate via a support bar arrangement that suspends the integrated circuit over the substrate. The creation of this gap functions to release the device from the substrate and create a thermally-isolated integrated circuit.

  11. Method of making thermally-isolated silicon-based integrated circuits

    Science.gov (United States)

    Wojciechowski, Kenneth; Olsson, Roy; Clews, Peggy J.; Bauer, Todd

    2017-11-21

    Thermally isolated devices may be formed by performing a series of etches on a silicon-based substrate. As a result of the series of etches, silicon material may be removed from underneath a region of an integrated circuit (IC). The removal of the silicon material from underneath the IC forms a gap between remaining substrate and the integrated circuit, though the integrated circuit remains connected to the substrate via a support bar arrangement that suspends the integrated circuit over the substrate. The creation of this gap functions to release the device from the substrate and create a thermally-isolated integrated circuit.

  12. Hanford Site Permanent Isolation Surface Barrier Development Program: Fiscal year 1992 and 1993 highlights

    International Nuclear Information System (INIS)

    Cadwell, L.L.; Link, S.O.; Gee, G.W.

    1993-09-01

    The Hanford Site Permanent Isolation Surface Barrier Development Program was jointly developed by the Pacific Northwest Laboratory and Westinghouse Hanford Company to design and test an earthen cover system that can be used to inhibit water infiltration; plant, animal, and human intrusion; and wind and water erosion. Kaiser Engineers Hanford Company provided engineering design support for the program. Work on barrier design has been under way at Hanford for nearly 10 years. The comprehensive development of a long-term barrier, formerly the Hanford Site Protective Barrier Development Program, was initiated in FY 1986, and a general field-tested design is expected to be completed by FY 1998. Highlights of efforts in FY 1992 and FY 1993 included the resumption of field testing, the completion of the prototype barrier design, and the convening of an external peer review panel, which met twice with the barrier development team. The review panel provided helpful guidance on current and future barrier development activities, while commending the program for its significant technical contributions to innovative barrier technology development

  13. Identification of Candida albicans and Candida dubliniensis Species Isolated from Bronchoalveolar Lavage Samples Using Genotypic and Phenotypic Methods

    Directory of Open Access Journals (Sweden)

    Sahar Kianipour

    2018-01-01

    Full Text Available Background: Candida dubliniensis is a newly diagnosed species very similar to Candida albicans phenotypically and first discovered in the mouth of people with AIDS in 1995. Among the different phenotypic and genotypic methods, a cost-effective method should be selected which makes it possible to differentiate these similar species. Materials and Methods: Polymerase chain reaction (PCR-restriction fragment length polymorphism with MspI enzyme and the Duplex-PCR method were done by DNA extraction using boiling. The sequencing of the amplified ribosomal region was used to confirm the C. dubliniensis species. Direct examination and colony count of the yeasts were applied for bronchoalveolar lavage (BAL samples and the growth rate of the yeasts were studied at 45°C. To understand the ability formation of chlamydoconidia in yeast isolates, they were separately cultured on the sunflower seed agar, wheat flour agar, and corn meal agar media. Results: Fifty-nine (49.2% yeast colonies were identified from the total of 120 BAL specimens. Twenty-nine isolated yeasts; including 17 (58.6% of C. albicans/dubliniensis complex and 12 (41.4% of nonalbicans isolates produced pseudohypha or blastoconidia in direct smear with a mean colony count of 42000 CFU/mL. C. albicans with the frequency of 15 (42.9% were the most common isolated yeasts, whereas C. dubliniensis was identified in two nonHIV patients. Conclusion: Sequencing of the replicated gene fragment is the best method for identifying the yeasts, but the determination of the species by phenotypic methods such as the creation of chlamydoconidia in sunflower seeds agar and wheat flour agar media can be cost-effective, have sensitivity and acceptable quality.

  14. Developing methods for detecting radioactive scrap

    International Nuclear Information System (INIS)

    Bellian, J.G.; Johnston, J.G.

    1995-01-01

    During the last 10 years, there have been major developments in radiation detection systems used for catching shielded radioactive sources in scrap metal. The original testing required to determine the extent of the problem and the preliminary designs of the first instruments will be discussed. Present systems available today will be described listing their advantages and disadvantages. In conclusion, the newest developments and state of the art equipment will also be included describing the limits and most appropriate locations for the systems

  15. The Role of Isolation Methods on a Nanoscale Surface Structure and Its Effect on the Size of Exosomes

    Directory of Open Access Journals (Sweden)

    JungReem Woo

    2016-06-01

    Full Text Available Exosomes are ~100 nanometre diameter vesicles secreted by mammalian cells. These emerging disease biomarkers carry nucleic acids, proteins and lipids specific to the parental cells that secrete them. Exosomes are typically isolated in bulk by ultracentrifugation, filtration or immu‐ noaffinity precipitation for downstream proteomic, genomic, or lipidomic analysis. However, the structural properties and heterogeneity of isolated exosomes at the single vesicle level are not well characterized due to their small size. In this paper, by using high-resolution atomic force microscope imaging, we show the nanoscale mor‐ phology and structural heterogeneity in exosomes derived from U87 cells. Quantitative assessment of single exosomes reveals nanoscale variations in morphology, surface roughness and counts isolated by ultracentrifugation (UC and immunoaffinity (IA purification. Both methods produce intact globular, 30-120 nm sized vesicles when imaged under fluid and in air. However, IA exosomes had higher surface roughness and bimodal size population compared to UC exosomes. The study highlights the differences in size and surface topography of exosomes purified from a single cell type using different isolation methods.

  16. A method for the isolation and characterization of functional murine monoclonal antibodies by single B cell cloning.

    Science.gov (United States)

    Carbonetti, Sara; Oliver, Brian G; Vigdorovich, Vladimir; Dambrauskas, Nicholas; Sack, Brandon; Bergl, Emilee; Kappe, Stefan H I; Sather, D Noah

    2017-09-01

    Monoclonal antibody technologies have enabled dramatic advances in immunology, the study of infectious disease, and modern medicine over the past 40years. However, many monoclonal antibody discovery procedures are labor- and time-intensive, low efficiency, and expensive. Here we describe an optimized mAb discovery platform for the rapid and efficient isolation, cloning and characterization of monoclonal antibodies in murine systems. In this platform, antigen-binding splenic B cells from immunized mice are isolated by FACS and cocultured with CD40L positive cells to induce proliferation and mAb production. After 12days of coculture, cell culture supernatants are screened for antigen, and IgG positivity and RNA is isolated for reverse-transcription. Positive-well cDNA is then amplified by PCR and the resulting amplicons can be cloned into ligation-independent expression vectors, which are then used directly to transfect HEK293 cells for recombinant antibody production. After 4days of growth, conditioned medium can be screened using biolayer interferometry for antigen binding and affinity measurements. Using this method, we were able to isolate six unique, functional monoclonal antibodies against an antigen of the human malaria parasite Plasmodium falciparum. Importantly, this method incorporates several important advances that circumvent the need for single-cell PCR, restriction cloning, and large scale protein production, and can be applied to a wide array of protein antigens. Copyright © 2017 Elsevier B.V. All rights reserved.

  17. Agile Service Development: A Rule-Based Method Engineering Approach

    NARCIS (Netherlands)

    dr. Martijn Zoet; Stijn Hoppenbrouwers; Inge van de Weerd; Johan Versendaal

    2011-01-01

    Agile software development has evolved into an increasingly mature software development approach and has been applied successfully in many software vendors’ development departments. In this position paper, we address the broader agile service development. Based on method engineering principles we

  18. A rapid cleanup method for the isolation and concentration of pyrrolizidine alkaloids in comfrey root.

    Science.gov (United States)

    Gray, Dean E; Porter, Andrew; O'Neill, Terry; Harris, Roger K; Rottinghaus, George E

    2004-01-01

    Preparations from comfrey (Symphytum officinale and S. x uplandicum) root and leaf contain varying levels of the hepatotoxic pyrrolizidine alkaloids (PAs). Reference compounds for comfrey are not commercially available, and there is currently no rapid extraction or analytical method capable of determining low levels in raw materials or as adulterants in commercially available extracts. A solid-phase extraction (SPE) method was developed using an Ergosil cleanup column that specifically binds the PAs. With this method, powdered comfrey root was extracted by sonication and shaking with basic chloroform. The extract was applied to the cleanup column under vacuum, washed with 2 mL acetone-chloroform (8 + 2, v/v) followed by 2 mL petroleum ether to remove excess chloroform. The column was dried under vacuum, and the PAs were eluted with 2 successive 1 mL aliquots methanol. Percent recoveries of the PAs following Ergosil SPE had an overall average of 96.8%, with RSD of 3.8% over a range of 1.0 to 25.0 g extracted in 100 mL. Average precision of the method (n = 3 over 4 extraction concentrations) gave an overall RSD of 6.0% for the 5 alkaloids, with a range of 0.8% (5 g in 100 mL) to 11.2% (25 g in 100 mL). Recovery optimization testing showed that 1.0 g comfrey root extracted in 100 mL yielded the greatest recovery (% dry weight) of the PAs, with an extraction efficiency and accuracy of 94.2%, and RSD of 1.7% (n = 9). The unique properties of the Ergosil cleanup column provide rapid sample cleanup, volume reduction, and concentration of PAs from comfrey extracts, and allow the eluant to be analyzed directly by traditional chromatographic methods.

  19. Development of three dimensional seismic isolation device with laminated rubber bearing and rolling seal type air spring

    International Nuclear Information System (INIS)

    Okada, Yasuo; Suhara, Junji; Tamura, Tadashi; Ohta, Kazuya; Moro, Satoshi

    2003-01-01

    Three dimensional (3D) seismic isolation device has been developed to use for the base isolation system of the heavy building like a nuclear reactor building. The developed device is the 3D seismic isolation device that consists of the laminated rubber baring as a horizontal isolation device and the rolling seal type air spring as the vertical isolation device in series. In this research, the 3D seismic isolation device reduction model whose scale is 1/10 is made and the workability of the device by the horizontal and vertical dynamic load is examined. Two experiment parameters are considered. One is the case that the structure of the part that the horizontal load and the vertical load contact is pin condition and the other is the case of the roller condition. As a result of the examination, the workability of the vertical direction is confirmed when the horizontal load acts. The pressure resistant ability test for the air spring is performed by the monotonic pressurization. As the result, it is confirmed that pressure resistant ability improved by restricting the side deformation of the air spring and that the material of the existing air spring can withstand high pressure use sufficiently. As the result, it is confirmed that the developed 3D seismic isolation device is applicable to the actual plant. This study was performed under the sponsorship of the Ministry of Economy, Trade and Industry of Japan. (author)

  20. Development of New Bituminous Pavement Design Method

    DEFF Research Database (Denmark)

    Ullidtz, Per

    The report and work of COST Action 333 sets in place the foundation for a coherent, cost-effective and harmonised European pavement design method. In order to do this, the work programme focused on information gathering, identification of requirements and the selection of the necessary design...

  1. Development of an indirect spectrophotometric method for ...

    African Journals Online (AJOL)

    A simple and rapid indirect spectrophotometric method for determination of ... of the colored product was measured at 405 nm and pH 3 against a reagent blank. ... The limit of detection and quantification were found to be 0.20±0.03 and ...

  2. Development of Quantification Method for Bioluminescence Imaging

    International Nuclear Information System (INIS)

    Kim, Hyeon Sik; Min, Jung Joon; Lee, Byeong Il; Choi, Eun Seo; Tak, Yoon O; Choi, Heung Kook; Lee, Ju Young

    2009-01-01

    Optical molecular luminescence imaging is widely used for detection and imaging of bio-photons emitted by luminescent luciferase activation. The measured photons in this method provide the degree of molecular alteration or cell numbers with the advantage of high signal-to-noise ratio. To extract useful information from the measured results, the analysis based on a proper quantification method is necessary. In this research, we propose a quantification method presenting linear response of measured light signal to measurement time. We detected the luminescence signal by using lab-made optical imaging equipment of animal light imaging system (ALIS) and different two kinds of light sources. One is three bacterial light-emitting sources containing different number of bacteria. The other is three different non-bacterial light sources emitting very weak light. By using the concept of the candela and the flux, we could derive simplified linear quantification formula. After experimentally measuring light intensity, the data was processed with the proposed quantification function. We could obtain linear response of photon counts to measurement time by applying the pre-determined quantification function. The ratio of the re-calculated photon counts and measurement time present a constant value although different light source was applied. The quantification function for linear response could be applicable to the standard quantification process. The proposed method could be used for the exact quantitative analysis in various light imaging equipment with presenting linear response behavior of constant light emitting sources to measurement time

  3. Blue ghosts: a new method for isolating amber mutants defective in essential genes of Escherichia coli

    DEFF Research Database (Denmark)

    Brown, S; Brickman, E R; Beckwith, J

    1981-01-01

    We describe a technique which permits an easy screening for amber mutants defective in essential genes of Escherichia coli. Using this approach, we have isolated three amber mutants defective in the rho gene. An extension of the technique allows the detection of ochre mutants and transposon inser...

  4. Hydrologic connectivity between geographically isolated wetlands and surface water systems: A review of select modeling methods

    Science.gov (United States)

    Heather E. Golden; Charles R. Lane; Devendra M. Amatya; Karl W. Bandilla; Hadas Raanan Kiperwas Kiperwas; Christopher D. Knightes; Herbert. Ssegane

    2014-01-01

    Geographically isolated wetlands (GIW), depressional landscape features entirely surrounded by upland areas, provide a wide range of ecological functions and ecosystem services for human well-being. Current and future ecosystem management and decision-making rely on a solid scientific understanding of how hydrologic processes affect these important GIW services and...

  5. A Novel Method for Isolation of Pluripotent Stem Cells from Human Umbilical Cord Blood.

    Science.gov (United States)

    Monti, Manuela; Imberti, Barbara; Bianchi, Niccolò; Pezzotta, Anna; Morigi, Marina; Del Fante, Claudia; Redi, Carlo Alberto; Perotti, Cesare

    2017-09-01

    Very small embryonic-like cells (VSELs) are a population of very rare pluripotent stem cells isolated in adult murine bone marrow and many other tissues and organs, including umbilical cord blood (UCB). VSEL existence is still not universally accepted by the scientific community, so for this purpose, we sought to investigate whether presumptive VSELs (pVSELs) could be isolated from human UCB with an improved protocol based on the isolation of enriched progenitor cells by depletion of nonprogenitor cells with magnetic separation. Progenitor cells, likely including VSELs, cultured with retinoic acid were able to form dense colonies and cystic embryoid bodies and to differentiate toward the ecto-meso-endoderm lineages as shown by the positivity to specific markers. VSEL differentiative potential toward mesodermal lineage was further demonstrated in vitro upon exposure to an established inductive protocol, which induced the acquisition of renal progenitor cell phenotype. VSEL-derived renal progenitors showed regenerative potential in a cisplatin model of acute kidney injury by restoring renal function and tubular structure through induction of proliferation of endogenous renal cells. The data presented here foster the great debate that surrounds VSELs and, more in general, the existence of cells endowed with pluripotent features in adult tissues. In fact, the possibility to find and isolate subpopulations of cells that fully fit all the criteria utilized to define pluripotency remains, nowadays, almost unproven. Thus, efforts to better characterize the phenotype of these intriguing cells are crucial to understand their possible applications for regenerative and precision medicine purposes.

  6. New method of plant mitochondria isolation and sub-fractionation for proteomic analyses

    Czech Academy of Sciences Publication Activity Database

    Hájek, Tomáš; Honys, David; Čapková, Věra

    2004-01-01

    Roč. 167, č. 3 (2004), s. 389-395 ISSN 0168-9452 R&D Projects: GA MŠk LZ1K03018 Institutional research plan: CEZ:AV0Z5038910 Keywords : plant mitochondria isolation * sub-fractionation * protein analysis Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 1.389, year: 2004

  7. Optimization of HPLC method for the isolation of Hypericum perforatum L. methanol extract

    Directory of Open Access Journals (Sweden)

    Stamenković, J.

    2013-12-01

    Full Text Available St. John's Wort (Hypericum perforatum L. is one of the most studied plant species in the family Hypericaceae. The aim of this study was the identification of the constituents of methanol extract of H. perforatum and optimization of conditions for their isolation. The main components of the methanol extract were isolated on preparative ZORBAX Eclipse XDB C18 column with solvent system consisting of methanol and 1x10-2 M ammonium acetate in water. Constituents of the extract were identified by comparing their retention times with the retention times of the standards, with the literature data and the UV spectra. By varying the conditions of chromatography, the optimal conditions for isolation of the methanol extract constituents were determined: mobile phase consisting of methanol and 1x10-2 M ammonium acetate in water in ratio 1 : 1, sample concentration 100 mg/mL, sample volume 30 µL, flow 2 mL/min. Under these conditions 7 components of the methanol extract were isolated.

  8. Identification of Candida albicans and Candida dubliniensis Species Isolated from Bronchoalveolar Lavage Samples Using Genotypic and Phenotypic Methods.

    Science.gov (United States)

    Kianipour, Sahar; Ardestani, Mohammad Emami; Dehghan, Parvin

    2018-01-01

    Candida dubliniensis is a newly diagnosed species very similar to Candida albicans phenotypically and first discovered in the mouth of people with AIDS in 1995. Among the different phenotypic and genotypic methods, a cost-effective method should be selected which makes it possible to differentiate these similar species. Polymerase chain reaction (PCR)-restriction fragment length polymorphism with MspI enzyme and the Duplex-PCR method were done by DNA extraction using boiling. The sequencing of the amplified ribosomal region was used to confirm the C. dubliniensis species. Direct examination and colony count of the yeasts were applied for bronchoalveolar lavage (BAL) samples and the growth rate of the yeasts were studied at 45°C. To understand the ability formation of chlamydoconidia in yeast isolates, they were separately cultured on the sunflower seed agar, wheat flour agar, and corn meal agar media. Fifty-nine (49.2%) yeast colonies were identified from the total of 120 BAL specimens. Twenty-nine isolated yeasts; including 17 (58.6%) of C. albicans / dubliniensis complex and 12 (41.4%) of nonalbicans isolates produced pseudohypha or blastoconidia in direct smear with a mean colony count of 42000 CFU/mL. C. albicans with the frequency of 15 (42.9%) were the most common isolated yeasts, whereas C. dubliniensis was identified in two nonHIV patients. Sequencing of the replicated gene fragment is the best method for identifying the yeasts, but the determination of the species by phenotypic methods such as the creation of chlamydoconidia in sunflower seeds agar and wheat flour agar media can be cost-effective, have sensitivity and acceptable quality.

  9. Technology development of p-type microstrip detectors with radiation hard p-spray isolation

    International Nuclear Information System (INIS)

    Pellegrini, G.; Fleta, C.; Campabadal, F.; Diez, S.; Lozano, M.; Rafi, J.M.; Ullan, M.

    2006-01-01

    A technology for the fabrication of p-type microstrip silicon radiation detectors using p-spray implant isolation has been developed at CNM-IMB. The p-spray isolation has been optimized in order to withstand a gamma irradiation dose up to 50 Mrad (Si), which represents the ionization radiation dose expected in the middle region of the SCT-Atlas detector of the future Super-LHC during 10 years of operation. The best technological options for the p-spray implant were found by using a simulation software package and dedicated calibration runs. Using the optimized technology, detectors have been fabricated in the Clean Room facility of CNM-IMB, and characterized by reverse current and capacitance measurements before and after irradiation. The average full depletion voltage measured on the non-irradiated detectors was V FD =41±3 V, while the leakage current density for the microstrip devices at V FD +20 V was 400 nA/cm 2

  10. DEVELOPMENT OF PIGEON PEA INOCULATED WITH RHIZOBIUM ISOLATED FROM COWPEA TRAP HOST PLANTS

    Directory of Open Access Journals (Sweden)

    SALOMÃO LIMA GUIMARÃES

    2016-01-01

    Full Text Available Pigeon pea is an important protein source grown in several tropical and sub - tropical countries, and is considered a multi - purpose plant that is resistant to the conditions of the Brazilian Cerrado. Among the possible uses for cowpea, its use as a green manure, increasing soil nitrogen content through the association with diazotrophic bacteria, generically known as rhizobia, is noteworthy. The present work aimed to evaluate the efficiency of Rhizobium strains isolated from cowpea plants in the development of pigeon peas cultured in Red Latosol. The experiment was conducted in a greenhouse, using a completely randomized design with seven treatments and four replications. Treatments consisted of inoculation with four Rhizobium strains (MT8, MT15, MT16, and MT23 and one commercial inoculant comprising Bradyrhizobium spp. strains BR 2801 and BR 2003. There were two controls, one absolute (without inoculation or nitrogen fertilization and the other with nitrogen fertilization. Each experimental plot consisted of an 8 - dm 3 vase containing three plants. Analyzed variables included plant height, SPAD index, number and dry weight of nodules, and shoot and root dry masses. Pigeon peas responded significantly to inoculation treatment, since all the plants inoculated with Rhizobium strains isolated from cowpea strains showed results similar to plants in the nitrogen control and commercial inoculant treatments. This demonstrates a favorable plant – bacteria interaction, which can be utilized as an alternative nitrogen source for pigeon peas.

  11. Current status of fluoride volatility method development

    Energy Technology Data Exchange (ETDEWEB)

    Uhlir, J.; Marecek, M.; Skarohlid, J. [UJV - Nuclear Research Institute, Research Centre Rez, CZ-250 68 Husinec - Rez 130 (Czech Republic)

    2013-07-01

    The Fluoride Volatility Method is based on a separation process, which comes out from the specific property of uranium, neptunium and plutonium to form volatile hexafluorides whereas most of fission products (mainly lanthanides) and higher transplutonium elements (americium, curium) present in irradiated fuel form nonvolatile tri-fluorides. Fluoride Volatility Method itself is based on direct fluorination of the spent fuel, but before the fluorination step, the removal of cladding material and subsequent transformation of the fuel into a powdered form with a suitable grain size have to be done. The fluorination is made with fluorine gas in a flame fluorination reactor, where the volatile fluorides (mostly UF{sub 6}) are separated from the non-volatile ones (trivalent minor actinides and majority of fission products). The subsequent operations necessary for partitioning of volatile fluorides are the condensation and evaporation of volatile fluorides, the thermal decomposition of PuF{sub 6} and the finally distillation and sorption used for the purification of uranium product. The Fluoride Volatility Method is considered to be a promising advanced pyrochemical reprocessing technology, which can mainly be used for the reprocessing of oxide spent fuels coming from future GEN IV fast reactors.

  12. Developing methods of controlling quality costs

    Directory of Open Access Journals (Sweden)

    Gorbunova A. V.

    2017-01-01

    Full Text Available The article examines issues of managing quality costs, problems of applying economic methods of quality control, implementation of progressive methods of quality costs management in enterprises with the view of improving the efficiency of their evaluation and analysis. With the aim of increasing the effectiveness of the cost management mechanism, authors introduce controlling as a tool of deviation analysis from the standpoint of the process approach. A list of processes and corresponding evaluation criteria in the quality management system of enterprises is introduced. Authors also introduce the method of controlling quality costs and propose it for the practical application, which allows them to determine useful and unnecessary costs at the existing operating plant. Implementing the proposed recommendations in the system of cost management at an enterprise will allow to improve productivity of processes operating and reduce wasted expense on the quality of the process on the basis of determining values of useful and useless costs of quality according to criteria of processes functioning in the system of quality management.

  13. Child Development in Developing Countries: Introduction and Methods

    Science.gov (United States)

    Bornstein, Marc H.; Britto, Pia Rebello; Nonoyama-Tarumi, Yuko; Ota, Yumiko; Petrovic, Oliver; Putnick, Diane L.

    2012-01-01

    The Multiple Indicator Cluster Survey (MICS) is a nationally representative, internationally comparable household survey implemented to examine protective and risk factors of child development in developing countries around the world. This introduction describes the conceptual framework, nature of the MICS3, and general analytic plan of articles…

  14. Development of standard testing methods for nuclear-waste forms

    International Nuclear Information System (INIS)

    Mendel, J.E.; Nelson, R.D.

    1981-11-01

    Standard test methods for waste package component development and design, safety analyses, and licensing are being developed for the Nuclear Waste Materials Handbook. This paper describes mainly the testing methods for obtaining waste form materials data

  15. Development of a high performance liquid chromatography method ...

    African Journals Online (AJOL)

    Development of a high performance liquid chromatography method for simultaneous ... Purpose: To develop and validate a new low-cost high performance liquid chromatography (HPLC) method for ..... Several papers have reported the use of ...

  16. DEVELOPMENT OF A MULTIPLE-LOCUS VARIABLE NUMBER OF TANDEM REPEAT ANALYSIS (MLVA FOR HELICOBACTER PYLORI AND ITS APPLICATION TO HELICOBACTER PYLORI ISOLATES FROM ROSTOV REGION,RUSSIA

    Directory of Open Access Journals (Sweden)

    Sorokin VM

    2012-09-01

    Full Text Available Stomach infection with Helicobacter pylori (H. pylori is the second most common infectious disease of humans. The severe pathological consequences of this infection include gastric and duodenal ulcer disease, the development of gastric mucosal atrophy, gastric carcinoma, and, more rarely, malignant tumors of the lymphoma. H. pylori infections cause very high morbidity and mortality and are of particular concern in developing countries, where H. pylori prevalences as high as 90% have been reported. The population of H. pylori shows a high genomic variability among isolates. And the polymorphism of repeat-units of genomics had participated the important process of evolution. A variety of molecular typing tools have been developed to access genetic relatedness in H. pylori isolates. However, there is still no standard genotyping system of this bacterium. The MLVA (Multi-Locus of Variable number of tandem repeat Analysis method is useful for performing phylogenetic analysis and is widely used in bacteria genotyping; however, there's little application in H. pylori analysis. This article is the first application of the MLVA method to investigate H. pylori isolates in Russia. MLVA of 4 VNTR loci with high discrimination power based on 10 candidates were performed on a collection of 22 strains of H. pylori which originated from Rostov region of Russia. This method provides a starting point on which improvements to the method and comparisons to other techniques can be made.

  17. Evaluation of different continuous cell lines in the isolation of mumps virus by the shell vial method from clinical samples

    Science.gov (United States)

    Reina, J; Ballesteros, F; Mari, M; Munar, M

    2001-01-01

    Aims—To compare prospectively the efficacy of the Vero, LLC-MK2, MDCK, Hep-2, and MRC-5 cell lines in the isolation of the mumps virus from clinical samples by means of the shell vial method. Methods—During an epidemic outbreak of parotiditis 48 clinical samples (saliva swabs and CSF) were studied. Two vials of the Vero, LLC-MK2, MDCK, MRC-5, and Hep-2 cell lines were inoculated with 0.2 ml of the samples by the shell vial assay. The vials were incubated at 36°C for two and five days. The vials were then fixed with acetone at -20°C for 10 minutes and stained by a monoclonal antibody against mumps virus by means of an indirect immunofluorescence assay. Results—The mumps virus was isolated from 36 samples. The Vero and LLC-MK2 cell lines showed a 100% isolation capacity, MDCK showed 77.7%, MRC-5 showed 44.4%, and Hep-2 showed 22.2%. The Vero and LLC-MK2 lines were significantly different to the other cell lines (p 5 infectious foci) were 94.4% for Vero, 97.2% for LLC-MK2, 5.5% for MDCK, 5.5% for Hep-2, and 0% for MRC-5. Conclusions—The Vero and LLC-MK2 cell lines are equally efficient at two and five days incubation for the isolation of the mumps virus from clinical samples, and the use of the shell vial method considerably shortens the time of aetiological diagnosis with higher specificity. Key Words: mumps virus • Vero cell line • LLC-MK2 cell line • MDCK cell line • Hep-2 cell line • MRC-5 cell line • isolation • shell vial PMID:11729211

  18. Comparison of Total RNA Isolation Methods for Analysis of Immune-Related microRNAs in Market Milks.

    Science.gov (United States)

    Oh, Sangnam; Park, Mi Ri; Son, Seok Jun; Kim, Younghoon

    2015-01-01

    Bovine milk provides essential nutrients, including immunologically important molecules, as the primary source of nutrition to newborns. Recent studies showed that RNAs from bovine milk contain immune-related microRNAs (miRNA) that regulate various immune systems. To evaluate the biological and immunological activity of miRNAs from milk products, isolation methods need to be established. Six methods for extracting total RNAs from bovine colostrums were adopted to evaluate the isolating efficiency and expression of miRNAs. Total RNA from milk was presented in formulation of small RNAs, rather than ribosomal RNAs. Column-combined phenol isolating methods showed high recovery of total RNAs, especially the commercial columns for biofluid samples, which demonstrated outstanding efficiency for recovering miRNAs. We also evaluated the quantity of five immune-related miRNAs (miR-93, miR-106a, miR-155, miR-181a, miR-451) in milk processed by temperature treatments including low temperature for long time (LTLT, 63℃ for 30 min)-, high temperature for short time (HTST, 75℃ for 15 s)-, and ultra heat treatment (UHT, 120-130℃ for 0.5-4 s). All targeted miRNAs had significantly reduced levels in processed milks compared to colostrum and raw mature milk. Interestingly, the amount of immune-related miRNAs from HTST milk was more resistant than those of LTLT and UHT milks. Our present study examined defined methods of RNA isolation and quantification of immune-specific miRNAs from small volumes of milk for use in further analysis.

  19. Prevalence of ESBL-producing Pseudomonas aeruginosa isolates in Warsaw, Poland, detected by various phenotypic and genotypic methods.

    Directory of Open Access Journals (Sweden)

    Agnieszka E Laudy

    Full Text Available Knowledge of the prevalence of ESBL enzymes among P. aeruginosa strains compared to the Enterobacteraiceae family is limited. The phenotypic tests recommended by EUCAST for the detection of ESBL-producing Enterobacteriaceae are not always suited for P. aeruginosa strains. This is mainly due to the presence of other families of ESBLs in P. aeruginosa isolates more often than in Enterobacteriaceae, production of natural AmpC cephalosporinase and its overexpression, and co-production of metallo-β-lactamases. The aim of this study was to determine the occurrence of ESBLs in P. aeruginosa isolated from patients from hospitals in Warsaw, to evaluate the ESBL production of these isolates using currently available phenotypic tests, their modifications, multiplex PCR and molecular typing of ESBL-positive isolates by PFGE. Clinical isolates of P. aeruginosa were collected in 2000-2014 from four Warsaw hospitals. Based on the data obtained in this study, we suggest using three DDST methods with inhibitors, such as clavulanic acid, sulbactam and imipenem, to detect ESBL-producing P. aeruginosa strains. Depending on the appearance of the plates, we suggest a reduction in the distance between discs with antibiotics to 15 mm and the addition of boronic acid at 0.4 mg per disc. The analysed isolates carried genes encoding ESBL from the families VEB (69 isolates with VEB-9, GES (6 with GES-1, 1 GES-5, 5 GES-13 and 2 with GES-15, OXA-2 (12 with OXA-15, 1 OXA-141, 1 OXA-210, 1 OXA-543 and 1 with OXA-544 and OXA-10 (5 isolates with OXA-74 and one with OXA-142. The most important result of this study was the discovery of three new genes, blaGES-15, blaOXA-141 and blaOXA-142; their nucleotide sequences have been submitted to the NCBI GenBank. It is also very important to note that this is the first report on the epidemiological problem of VEB-9-producing bacterial strains, not only in Poland but also worldwide.

  20. Development and Validation of a Dissolution Test Method for ...

    African Journals Online (AJOL)

    Purpose: To develop and validate a dissolution test method for dissolution release of artemether and lumefantrine from tablets. Methods: A single dissolution method for evaluating the in vitro release of artemether and lumefantrine from tablets was developed and validated. The method comprised of a dissolution medium of ...

  1. Feedback of reactor operating data to nuclear methods development

    International Nuclear Information System (INIS)

    Crowther, R.L.; Kang, C.M.; Parkos, G.R.; Wolters, R.A.

    1978-01-01

    The problems in obtaining power reactor data for reliable nuclear methods development and the major sources of power reactor data for this purpose are reviewed. Specific examples of the use of power reactor data in nuclear methods development are discussed. The paper concludes with recommendations on the key elements of an effective program to use power reactor data in nuclear methods development

  2. Energetic strategies for the sustainable development of the isolated areas of Amazon

    International Nuclear Information System (INIS)

    Di Lascio, M.A.

    1999-01-01

    This article analyzes the economic and energetic situation of the 35 % Amazonian inhabitants that live in remote and isolated sites, inside the Amazon forest. Generally, deforestation begins in those places and rapidly occurs the destruction of local ecosystems. The poverty is everywhere and the energy supply is insufficient or nonexistent. The electric energy is generated with diesel, but its acquisition consumes all available income.The man is not adapted to the environment, and destroys the forest to increase profit and comfort, but without protection the soil fragility causes poverty. To conclude some case studies are presented. Finally, it is proposed an energy strategy to protect the environment, through an an economic development, based on the use of renewable energy. (author)

  3. Data on isolating mesenchymal stromal cells from human adipose tissue using a collagenase-free method

    Directory of Open Access Journals (Sweden)

    Wassim Shebaby

    2016-03-01

    Full Text Available The present dataset describes a detailed protocol to isolate mesenchymal cells from human fat without the use of collagenase. Human fat specimen, surgically cleaned from non-fat tissues (e.g., blood vessels and reduced into smaller fat pieces of around 1–3 mm size, is incubated in complete culture media for five to seven days. Then, cells started to spread out from the fat explants and to grow in cultures according to an exponential pattern. Our data showed that primary mesenchymal cells presenting heterogeneous morphology start to acquire more homogenous fibroblastic-like shape when cultured for longer duration or when subcultured into new flasks. Cell isolation efficiency as well as cell doubling time were also calculated throughout the culturing experimentations and illustrated in a separate figure thereafter. This paper contains data previously considered as an alternative protocol to isolate adipose-derived mesenchymal stem cell published in “Proliferation and differentiation of human adipose-derived mesenchymal stem cells (ASCs into osteoblastic lineage are passage dependent” [1]. Keywords: Adipose tissue, mesenchymal stromal cell, cell culture, doubling time

  4. A Simple Response Evaluation Method for Base-Isolation Building-Connection Hybrid Structural System under Long-Period and Long-Duration Ground Motion

    Directory of Open Access Journals (Sweden)

    Kohei Hayashi

    2018-02-01

    Full Text Available An innovative hybrid control building system of base-isolation and building-connection has been proposed in the previous study. This system has two advantages, (i to resist an impulsive earthquake input through the base-isolation system and (ii to withstand a long-duration earthquake input through the building-connection system. A simple response evaluation method without the need of non-linear time–history response analysis is proposed here for this hybrid building system under a long-period and long-duration ground motion. An analytical expression is derived in the plastic deformation of an elastic–perfectly plastic single-degree-of-freedom (SDOF model with viscous damping under the multi-impulse, which is the representative of long-period and long-duration ground motions. A transformation procedure of a base-isolation building-connection hybrid structural system into an SDOF model is proposed by introducing two steps, one is the reduction of the main base-isolated building to an SDOF system, and the other is the reduction of the connecting oil dampers supported on a free-wall to an oil damper with a newly introduced compensation factor on a rigid wall. Application of the analytical expression of the plastic deformation to the reduced SDOF model including the compensation factor on the connecting oil dampers enables the development of a simplified, but rather accurate response evaluation method. The time–history response analysis of the multi-degree-of-freedom model and the comparison with the proposed simplified formula make clear the accuracy and reliability of the proposed simplified response evaluation method.

  5. Status and development of uranium prospection methods

    International Nuclear Information System (INIS)

    Barthel, F.

    1978-01-01

    In radiometric prospection, gamma measuring equipment is widely used. Simple instruments, so-called scintillometers, can only measure total radiation while spectrometers permit separate measurements of uranium, thorium, or calcium via daughter products of their decay chains. Depending on the target investigated, airborne, carborne, or footborne methods are employed. In radon prospection the gaseous decay product radon is measured as a sign of hidden uranium enrichment in ground air or water from springs. Due to its high solubility, uranium is well suited for geochemical prospection where uranium concentrations in bodies of water, river sediments, soil and rock types are determined. There is a trend in uranium prospection towards the discovery of hidden orifications. Novel techniques, e.g. airborne geochemistry, isotope chemistry, tracer element measurement, etc. are being tested with a view to their suitability for uranium prospection. (orig./HP) 891 HP/orig.- 892 MKO [de

  6. The association between WNT10A variants and dental development in patients with isolated oligodontia

    NARCIS (Netherlands)

    Dhamo, Brunilda; Fennis, Willem; Creton, Marijn; Vucic, Strahinja; Cune, Marco; van Amstel, Hans Kristian Ploos; Wolvius, Eppo B; van den Boogaard, Marie-Jose; Ongkosuwito, Edwin M.

    2017-01-01

    In this study we aimed to determine the effect of WNT10A variants on dental development in patients with oligodontia. Forty-three (25 boys and 18 girls) individuals were eligible for this study. Stage of development for each present tooth was assessed using the Demirjian method. In case no

  7. Effects of certain therapeutic factors on facial development in isolated cleft palate.

    Science.gov (United States)

    Smahel, Z

    1989-01-01

    Roentgencephalometry was used during the investigation of the effects of some therapeutic factors on the growth and development of the jaws in 64 adult males with an isolated cleft palate repaired by pushback. The anterior growth of the maxilla was not related to the age at the time of surgery or to orthodontic therapy with removable appliances. A small number of individuals operated during adolescence had also a shorter depth of the maxilla similarly as patients operated upon during early childhood. Anterior crossbite developed mostly in patients with reduced proclination of the upper alveolar process, while, on the contrary, a retrusion of the maxilla played no essential part. This observation proves useful for the prediction of the development of this malocclusion. The angle of sagittal jaw relations does not represent necessarily a valid criterion of the development of the jaws. In the presence of an overbite retrusion of the maxilla is associated with a retroposition of the mandible and thus the angle of sagittal jaw relations remains unchanged. Thus overbite represents an effective mechanism acting on the position of the mandible. A differentiated approach for the determination of the age of choice at the time of palate surgery according to the type and extent of the cleft is proposed.

  8. Hanford Site protective isolation surface barrier: Taking research and development to engineered application

    International Nuclear Information System (INIS)

    Myers, D.R.; Wing, N.R.

    1994-01-01

    The development of the Protective Isolation Surface Barrier has been an ongoing program since 1985. This development effort has focused on several technical areas. These technical areas include water infiltration, biointrusion, human intrusion, erosion/deposition, physical stability, barrier materials, computer modeling, long-term climate effects, natural analogs, and barrier design. This paper briefly reviews the results of the research and development in the technical areas and then explains how the results of this work have influenced the design features of the prototype barrier. A good example of this is to explain how the type and depth of the soil layer used in the barrier is related to water infiltration, biointrusion, modeling, climate, analogs, and barrier materials. Another good example is to explain the relationship of the barrier sideslopes (basalt riprap and native soil) with human intrusion, biointrusion, barrier materials, and barrier design. In general, the design features of the prototype barrier will be explained in terms of the results of the testing and development program. After the basis for prototype barrier design has been established, the paper will close by reviewing the construction of the prototype barrier, sharing the lessons learned during construction, and explaining the ongoing testing and monitoring program which will determine the success or failure of this barrier concept and the need for additional design modifications

  9. The long-term climate change task of the Hanford permanent isolation barrier development program

    International Nuclear Information System (INIS)

    1994-01-01

    The Hanford Site Permanent Isolation Barrier Development Program is developing an in-place disposal capability for low-level nuclear waste for the US Department of Energy at the Hanford Site in southeastern Washington State. Layered earthen and engineered barriers are being developed that will function in what is currently a semiarid environment (mean annual precipitation and temperature of 16 cm and 11.8 degrees C, respectively) for at least 1,000 yr by limiting the infiltration of water through the waste. The Long-Term Climate Change Task has specific goals of (1) obtaining defensible probabilistic projections of the long-term climate variability in the Hanford Site region at many different time scales into the future; (2) developing several test-case climate scenarios that bracket the range of potential future climate, including both greenhouse warming and cycling into another ice age; and (3) using the climate scenarios both to test and to model protective barrier performance. Results from the Carp Lake Pollen Coring Project indicate that for the last approximately 100,000 yr the Columbia River Basin's long-term range of mean annual precipitation ranged from 25%--50% below to 28% above modern levels, while temperature has ranged from 7 degrees C--10 degrees C below to 2 degrees C above modern levels. This long record provides confidence that such a range should bracket potential natural climate change even if the earth cycles back into another Ice Age in the next few millennia

  10. Detection of Antimicrobial Compounds Isolated from Several Tropical Lentinus by Bioautographic Method

    Directory of Open Access Journals (Sweden)

    LISDAR I. SUDIRMAN

    2005-06-01

    Full Text Available The antimicrobial compounds extracted either from culture filtrates or mycelia of several tropical Lentinus species could be detected their existences and locations by bioautographic method. For this purpose, the crude extracts were deposited as spots on silica gel plates and developed in a n-butanol-acetic acid-water mixture (3:1:1. The dry silica gel plates were then seeded with Bacillus subtilis and incubated at 35 oC for one night. On these plates, the extracts were separated into several bioautographic spots or growth inhibition zones. In parallel, the spots were detected by viewing with chemical revelations or under ultraviolet radiations at 254 nm or 366 nm. On silica gel thin-layer chromatograms, the crude extracts of Lentinus were separated into several bioautographic spots; for the filtrate extracts of L. squarrosulus 55A into three spots (Rfs 0.75, 0.50, 0.17, the mycelial extracts of L. sajor-caju LSC8 into two spots (Rfs 0.77, 0.54, the mycelial extract of L. torulosus LU3 into two spots (Rfs 0.77, 0.48, the filtrate extracts of L. cladopus LC6 into one spot (Rf 0.76 but the mycelial extracts of this mushroom separated into two spots (Rfs 0.79, 0.54, the filtrate and mycelial extracts of L. cladopus LC4 into three spots respectively (Rfs 0.75, 0.61, 0.45 for the filtrate extract and Rfs 0.83, 0.73, 0.60 for mycelial extract. By this method, the active compounds were detected directly and it is a usual method for further work on the purification of the target compounds.

  11. Transplantation of an alginate-matrigel matrix containing isolated ovarian stromal cells: first step in developing an artificial ovary

    OpenAIRE

    Vanacker, Julie; Dolmans, Marie-Madeleine; des Rieux, Anne; Jaeger, Jonathan; Luyckx, Valérie; Van Langendonckt, Anne; Donnez, Jacques; Andrade Amorim, Christiani; 3rd International Conference “Strategies in Tissue Engineering”

    2012-01-01

    Introduction: For women diagnosed with leukemia, transplantation of cryopreserved ovarian tissue after disease remission is not advisable due to the risk of reintroduction of malignant cells. To restore fertility in these patients, a biodegradable artificial ovary that offers an environment allowing isolated follicles and stromal cells (SCs) to survive and grow needs to be developed. The aim of this study is therefore to test the survival and proliferation of isolated SCs encapsulated in an a...

  12. Development of analytical methods for the gas chromatographic determination of 1,2-epoxy-3-butene, 1,2:3,4-diepoxybutane, 3-butene-1,2-diol, 3,4-epoxybutane-1,2-diol and crotonaldehyde from perfusate samples of 1,3-butadiene exposed isolated mouse and rat livers

    Energy Technology Data Exchange (ETDEWEB)

    Bhowmik, S.; Schuster, A.; Filser, J.G.

    2003-07-01

    Mutagenicity and carcinogenicity of 1,3-butadiene (BD) highly probably results from epoxide metabolites as 1,2-epoxy-3-butene (EB), 1,2:3,4-diepoxybutane (DEB) and 3,4-epoxybutane-1,2-diol (EBD). A further metabolite crotonaldehyde (CA) has also been discussed to be relevant. So far, in BD exposed rodents only EB and DEB concentrations had been quantified. However, the methods used were either not very sensitive or instrumentally expensive. Therefore, the goal of the present work was to establish simple analytical methods selective and sensitive enough to determine all of these compounds and a further secondary BD intermediate, 3-butene-1,2-diol (B-diol), in BD exposed rodent livers. The once-through perfused liver system was chosen for testing the applicability of the methods to be developed, since it enables BD exposures of this quantitatively most relevant metabolising organ near to the in-vivo situation. All the metabolites were extracted from the aqueous perfusion medium and analysed using a gas chromatograph equipped with a mass selective detector (GC/MS) in the PCI mode. (orig.)

  13. Development methods for VLSI-processors

    International Nuclear Information System (INIS)

    Horninger, K.; Sandweg, G.

    1982-01-01

    The aim of this project, which was originally planed for 3 years, was the development of modern system and circuit concepts, for VLSI-processors having a 32 bit wide data path. The result of this first years work is the concept of a general purpose processor. This processor is not only logically but also physically (on the chip) divided into four functional units: a microprogrammable instruction unit, an execution unit in slice technique, a fully associative cache memory and an I/O unit. For the ALU of the execution unit circuits in PLA and slice techniques have been realized. On the basis of regularity, area consumption and achievable performance the slice technique has been prefered. The designs utilize selftesting circuitry. (orig.) [de

  14. Methods and Protocols for Developing Prion Vaccines.

    Science.gov (United States)

    Marciniuk, Kristen; Taschuk, Ryan; Napper, Scott

    2016-01-01

    Prion diseases denote a distinct form of infectivity that is based in the misfolding of a self-protein (PrP(C)) into a pathological, infectious conformation (PrP(Sc)). Efforts to develop vaccines for prion diseases have been complicated by the potential dangers that are associated with induction of immune responses against a self-protein. As a consequence, there is considerable appeal for vaccines that specifically target the misfolded prion conformation. Such conformation-specific immunotherapy is made possible through the identification of vaccine targets (epitopes) that are exclusively presented as a consequence of misfolding. An immune response directed against these targets, termed disease-specific epitopes (DSEs), has the potential to spare the function of the native form of the protein while clearing, or neutralizing, the infectious isomer. Although identification of DSEs represents a critical first step in the induction of conformation-specific immune responses, substantial efforts are required to translate these targets into functional vaccines. Due to the poor immunogenicity that is inherent to self-proteins, and that is often associated with short peptides, substantial efforts are required to overcome tolerance-to-self and maximize the resultant immune response following DSE-based immunization. This often includes optimization of target sequences in terms of immunogenicity and development of effective formulation and delivery strategies for the associated peptides. Further, these vaccines must satisfy additional criteria from perspectives of specificity (PrP(C) vs. PrP(Sc)) and safety (antibody-induced template-driven misfolding of PrP(C)). The emphasis of this report is on the steps required to translate DSEs into prion vaccines and subsequent evaluation of the resulting immune responses.

  15. Developing methods for establishing improved plant reliability

    International Nuclear Information System (INIS)

    Zebroski, E.L.; Lapides, M.E.; Johnson, L.

    1976-01-01

    The current average capacity factor for nuclear electric generating facilities is approximately 60 percent, virtually the same as that for large, modern fossil-fired units. There is a clear cost advantage for nuclear electrical generation in comparison to coal-fired units at a common performance level. Because of the accelerated increase in capital and fuel costs of coal-fired units, the nuclear units will usually exhibit a cost advantage even when operating at lower capacity factor values. The incentives for improvement follow from the utility objectives to provide electricity at the most economical rates possible and are reflected in reduced system fuel costs and capital requirements. These objectives require maximizing the productivity of existing or planned investments in nuclear generation. The requirements are to: (1) achieve and sustain the highest capacity factor consistent with each utility's electric transmission system configuration and demand; (2) accelerate the rate of attainment of operational ''maturity'' of new generation; and (3) avoid low-probability but large-consequence forced-outage events. Nuclear unit productivity improvement involves utilities, regulatory agencies and industrial and government research and development organizations; the efforts appropriate to each organization were identified in a previous study. Since no foreseeable set of warranties begins to cover the consequential costs of low performance, the greatest incentive for improvement rests with each individual utility. The paper addresses the methodology and information sources for productivity actions by utilities during unit procurement and design phases

  16. Scenario development for the Waste Isolation Pilot Plant compliance certification application

    International Nuclear Information System (INIS)

    GALSON, D.A.; SWIFT, PETER N.; ANDERSON, D. RICHARD; BENNETT, D.G.

    1998-01-01

    Demonstrating compliance with the applicable regulations for the Waste Isolation Pilot Plant (WIPP) requires an assessment of the long-term performance of the disposal system. Scenario development is one starting point of this assessment, and generates inquiry about the present state and future evolution of the disposal system. Scenario development consists of four tasks: (1) identifying and classifying features, events and processes (FEPs), (2) screening FEPs according to well-defined criteria, (3) forming scenarios (combinations of FEPs) in the context of regulatory performance criteria and (4) specifying of scenarios for consequence analysis. The development and screening of a comprehensive FEP list provides assurance that the identification of significant processes and events is complete, that potential interactions between FEPs are not overlooked, and that responses to possible questions are available and well documented. Two basic scenarios have been identified for the WIPP: undisturbed performance (UP) and disturbed performance (DP). The UP scenario is used to evaluate compliance with the Environmental Protection Agency's (EPA's) Individual Dose (40 CFR Section 191-15) and Groundwater Protection (40 CFR Section 191-24) standards and accounts for all natural-, waste- and repository-induced FEPs that survive the screening process. The DP scenario is required for assessment calculations for the EPA's cumulative release standard (Containment Requirements, 40 CFR Section 191-13) and accounts for disruptive future human events, which have an uncertain probability of occurrence, in addition to the UP FEPs

  17. Characterization of competence and biofilm development of a Streptococcus sanguinis endocarditis isolate.

    Science.gov (United States)

    Zhu, L; Zhang, Y; Fan, J; Herzberg, M C; Kreth, J

    2011-04-01

    Streptococcus sanguinis is an oral commensal bacterium and endogenous pathogen in the blood, which is generally naturally competent to take up extracellular DNA. Regarded as a stress response, competence development enables S. sanguinis to acquire new genetic material. The sequenced reference strain SK36 encodes and expresses the genes required for competence (com) and uptake of DNA. Isolated from blood cultures of a confirmed case of infective endocarditis, strain 133-79 encodes all necessary com genes but is not transformable under conditions permissive for competence development in SK36. Using synthetic competence-stimulating peptides (sCSP) based on sequences of SK36 and 133-79 comC, both strains developed competence at similar frequencies in cross-transformation experiments. Furthermore, downstream response pathways are similar in strains SK36 and 133-79 because platelet aggregation and biofilm formation appeared unaffected by CSP. Collectively, the data indicate that strains SK36 and 133-79 respond to CSP similarly, strongly suggesting that endogenous production or release of CSP from 133-79 is impaired. © 2011 John Wiley & Sons A/S.

  18. Characterization of competence and biofilm development of a Streptocccus sanguinis endocarditis isolate

    Science.gov (United States)

    Zhu, Lin; Zhang, Yongshu; Fan, Jingyuan; Herzberg, Mark C.; Kreth, Jens

    2010-01-01

    Streptococcus sanguinis is an oral commensal bacterium and endogenous pathogen in the blood, which generally is naturally competent to take up extracellular DNA. Regarded as a stress response, competence development enables S. sanguinis to acquire new genetic material. The sequenced reference strain SK36 encodes and expresses the genes required for competence (com) and uptake of DNA. Isolated from blood cultures of a confirmed case of infective endocarditis, strain 133–79 encodes all necessary com genes but is not transformable under conditions permissive for competence development in SK36. Using synthetic competence-stimulating peptides (sCSP) based on sequences of SK36 and 133–79 comC, both strains developed competence at similar frequencies in cross-transformation experiments. Furthermore, downstream response pathways are similar in strains SK36 and 133–79 since platelet aggregation and biofilm formation appeared unaffected by CSP. Collectively, the data indicate that strains SK36 and 133–79 respond to CSP similarly, strongly suggesting that endogenous production or release of CSP from 133–79 is impaired. PMID:21375702

  19. Transplantation of an alginate-matrigel matrix containing isolated ovarian cells: first step in developing a biodegradable scaffold to transplant isolated preantral follicles and ovarian cells.

    Science.gov (United States)

    Vanacker, Julie; Luyckx, Valérie; Dolmans, Marie-Madeleine; Des Rieux, Anne; Jaeger, Jonathan; Van Langendonckt, Anne; Donnez, Jacques; Amorim, Christiani A

    2012-09-01

    For women diagnosed with leukemia, transplantation of cryopreserved ovarian tissue after disease remission is not advisable. Therefore, to restore fertility in these patients, we aim to develop a biodegradable artificial ovary that offers an environment where isolated follicles and ovarian cells (OCs) can survive and grow. Four NMRI mice were ovariectomized and their ovaries used to isolate OCs. Groups of 50,000 OCs were embedded in an alginate-matrigel matrix for further fixation (fresh controls), one week of in vitro culture (IVC) or heterotopic autografting. OC proliferation (Ki67), apoptosis (TUNEL), scaffold degradation, vessel formation (CD34) and inflammation (CD45) were analyzed. Ki67-positive OCs were found in 2.3%, 9.0% and 15.5% cells of cases in fresh, IVC and grafted beads respectively, while cells were TUNEL-positive in 0%, 1.5% and 6.9% of cases. After IVC or grafting, the beads degraded, losing their original round aspect, and infiltrating blood capillaries could be observed in the grafted beads. CD34-positive cells and 22% CD45-positive cells were found around and inside the matrix. In conclusion, our results demonstrate that an alginate-based matrix is a promising proposition to graft isolated OCs. After transplantation, this matrix was able to degrade, allowed vascularization and elicited a low inflammatory response. Copyright © 2012 Elsevier Ltd. All rights reserved.

  20. New method for laser driven ion acceleration with isolated, mass-limited targets

    International Nuclear Information System (INIS)

    Paasch-Colberg, T.; Sokollik, T.; Gorling, K.; Eichmann, U.; Steinke, S.; Schnuerer, M.; Nickles, P.V.; Andreev, A.; Sandner, W.

    2011-01-01

    A new technique to investigate laser driven ion acceleration with fully isolated, mass-limited glass spheres with a diameter down to 8μm is presented. A Paul trap was used to prepare a levitating glass sphere for the interaction with a laser pulse of relativistic intensity. Narrow-bandwidth energy spectra of protons and oxygen ions have been observed and were attributed to specific acceleration field dynamics in case of the spherical target geometry. A general limiting mechanism has been found that explains the experimentally observed ion energies for the mass-limited target.

  1. Development of safety evaluation guidelines for base-isolated buildings in Japan

    International Nuclear Information System (INIS)

    Aoyama, Hiroyuki

    1989-01-01

    This paper describes the safety evaluation guidelines and the review process for non-nuclear base-isolated buildings proposed for construction in Japan. The paper discusses the guidelines application for two types of soil: hard soil and intermediate soil (soft soil was excluded.); safety evaluation items included in the level C design review; and safety margin of base isolation. Lessons learned through these design review efforts have potential applicability to design of seismic base isolation for nuclear power plants

  2. The development of permanent isolation barriers for buried wastes in cool deserts: Hanford, Washington

    International Nuclear Information System (INIS)

    Link, S.O.; Gee, G.W.; Wing, N.R.

    1993-12-01

    The purpose of this report is to present the results of research on surface hydrology and the role of plants and animals on permanent isolation barrier effectiveness at Hanford. These topics are a subset of a larger set of studies on permanent isolation barriers. A complete review of these tasks has been documented. We also discuss current work that tests our integrated scientific and engineering concepts on a large prototype barrier to determine if it can isolate buried wastes from environmental dispersion

  3. Milestones in the Development of Iterative Solution Methods

    Directory of Open Access Journals (Sweden)

    Owe Axelsson

    2010-01-01

    Full Text Available Iterative solution methods to solve linear systems of equations were originally formulated as basic iteration methods of defect-correction type, commonly referred to as Richardson's iteration method. These methods developed further into various versions of splitting methods, including the successive overrelaxation (SOR method. Later, immensely important developments included convergence acceleration methods, such as the Chebyshev and conjugate gradient iteration methods and preconditioning methods of various forms. A major strive has been to find methods with a total computational complexity of optimal order, that is, proportional to the degrees of freedom involved in the equation. Methods that have turned out to have been particularly important for the further developments of linear equation solvers are surveyed. Some of them are presented in greater detail.

  4. A reliable combination method to identification and typing of epidemic and endemic clones among clinical isolates of Acinetobacter baumannii.

    Science.gov (United States)

    Piran, Arezoo; Shahcheraghi, Fereshteh; Solgi, Hamid; Rohani, Mahdi; Badmasti, Farzad

    2017-10-01

    The multi-drug resistant (MDR) Acinetobacter baumannii as an important nosocomial pathogen has emerged a global health concern in recent years. In this study, we applied three easier, faster, and cost-effective methods including PCR-based open reading frames (ORFs) typing, sequence typing of bla OXA-51-like and RAPD-PCR method to rapid typing of A. baumannii strains. Taken together in the present study the results of ORFs typing, PCR-sequencing of bla OXA-51-like genes and MLST sequence typing revealed there was a high prevalence (62%, 35/57) of ST2 as international and successful clone which detected among clinical isolates of multi-drug resistant A. baumannii with ORF pattern B and bla OXA-66 gene. Only 7% (4/57) of MDR isolates belonged to ST1 with ORF pattern A and bla OXA-69 gene. Interestingly, we detected singleton ST513 (32%, 18/57) that encoded bla OXA-90 and showed the ORF pattern H as previously isolated in Middle East. Moreover, our data showed RAPD-PCR method can detect divergent strains of the STs. The Cl-1, Cl-2, Cl-3, Cl-4, Cl-10, Cl-11, Cl-12, Cl-13 and Cl-14 belonged to ST2. While the Cl-6, Cl-7, Cl-8 and Cl-9 belonged to ST513. Only Cl-5 belonged to ST1. It seems that the combination of these methods have more discriminatory than any method separately and could be effectively applied to rapid detection of the clonal complex (CC) of A. baumannii strains without performing of MLST or PFGE. Copyright © 2017 Elsevier B.V. All rights reserved.

  5. Isolation of hydroxytyrosol from olive leaves extract, radioiodination and investigation of bioaffinity using in vivo/in vitro methods

    Energy Technology Data Exchange (ETDEWEB)

    Ozkan, M.; Biber Muftuler, F.Z.; Kilcar, A. Yurt; Medine, E.I.; Unak, P. [Ege Univ., Izmir (Turkey). Dept. of Nuclear Applications

    2013-11-01

    It is known that medicinal plants like olive have biological activities due to their flavonoid content such as olueropein, tyrosol, hydroxytyrosol etc. In current study, hydroxytrosol (HT) which is one of the major phenolic compounds in olive, olive leaves and olive oil, was isolated after methanol extraction and purification of olive leaves which are grown in the northern Anatolia region of Turkey. The isolated HT was radiolabeled with {sup 131}I ({sup 131}I-HT) and the bioaffinity of this radiolabeled component of olive leaves extract was investigated by using in vivo/in vitro methods. It was found that HT could be radiolabeled with {sup 131}I in yields of 95.6 {+-} 4.4% (n = 8), and in vivo studies showed that {sup 131}I-HT is taken up by urinary bladder, stomach, small intestine, large intestine, breast and prostate. Significant incorporation of activity was observed in cell lines via in vitro studies. (orig.)

  6. Isolation of Candida dubliniensis for the first time in Cali, Colombia, and its identification with phenotyping methods.

    Science.gov (United States)

    Alvarez, María Inés; Suárez, Blanca Lynne; Caicedo, Luz Dary

    2009-01-01

    Candida dubliniensis is an emerging pathogenic yeast isolated mainly from the oral cavity of HIV-infected patients. The close phenotypic and genotypic relationship between C. albicans and C. dubliniensis has led to incorrectly identifying isolates of C. dubliniensis as C. albicans. The oral cavities of 107 diabetic patients were studied in Cali, Colombia, and 72 colonies of Candida, with shades of green on CHROMagar Candida culture media, were obtained. Various phenotypic tests were carried out, which included germ tube formation and production of chlamydospores on corn meal Agar. Additionally, growth studies were carried out at 42 degrees C and 45 degrees C and on Sabouraud agar with 6.5%, sodium chloride. Identification of C. dubliniensis with these tests was confirmed with API 20C Aux. We identified 65 and 7 colonies of C. albicans and C. dubliniensis, respectively. This is the first time that C. dubliniensis is identified with phenotypic methods in Colombia.

  7. [Explantation method of isolating a persistent tick-borne encephalitis virus from the organs of infected monkeys].

    Science.gov (United States)

    Levina, L S; Pogodina, V V

    1981-01-01

    The method of explantation was used to examine 63 organs from M. rhesus monkeys 92-783 days after intracerebral and subcutaneous inoculation with the Vasilchenko, Aina/1448 and 41/65 strains of tick-borne encephalitis virus. The optimal time for examination of the explants by tests of the hemagglutinating, cytopathogenic activity of the virus and its pathogenicity for mice was found to be the 15th day of cultivation. A comparative study of the properties of 3 isolates obtained from explants of the spleen, liver and subcortical cerebral ganglia 202 and 307 days after inoculation of monkeys was carried out. The isolates differed from the parental TBE virus strains by their capacity to form small plaques in PEKV cell cultures (pig embryo kidney cells in versen medium).

  8. Comparison of two biochemical methods for identifying Corynebacterium pseudotuberculosis isolated from sheep and goats.

    Science.gov (United States)

    Huerta, Belén; Gómez-Gascón, Lidia; Vela, Ana I; Fernández-Garayzábal, José F; Casamayor, Almudena; Tarradas, Carmen; Maldonado, Alfonso

    2013-06-01

    The biochemical pattern of Cowan and Steel (BPCS) was compared with a commercial biochemical strip for the identification of Corynebacterium pseudotuberculosis isolated from small ruminants. On 16S rRNA gene sequencing, 40/78 coryneform isolates from the lymph nodes of sheep and goats with lesions resembling caseous lymphadenitis were identified as C. pseudotuberculosis. The sensitivities of the BPCS and the commercial biochemical strip relative to 16S rRNA sequencing were 80% and 85%, and their specificities were 92.1% and 94.7%, respectively; the level of agreement between the BPCS and the commercial biochemical strip was high (κ=0.82). Likelihood ratios for positive and negative results were 10.0 and 0.22 for the BPCS, and 16.0 and 0.16 for the commercial biochemical strip, respectively. These results indicate that the BPCS and the commercial biochemical strip are both useful for identifying C. pseudotuberculosis in veterinary microbiology laboratories. Copyright © 2012 Elsevier Ltd. All rights reserved.

  9. A Survey of Formal Methods in Software Development

    DEFF Research Database (Denmark)

    Bjørner, Dines

    2012-01-01

    The use of formal methods and formal techniques in industry is steadily growing. In this survey we shall characterise what we mean by software development and by a formal method; briefly overview a history of formal specification languages - some of which are: VDM (Vienna Development Method, 1974...... need for multi-language formalisation (Petri Nets, MSC, StateChart, Temporal Logics); the sociology of university and industry acceptance of formal methods; the inevitability of the use of formal software development methods; while referring to seminal monographs and textbooks on formal methods....

  10. Development of a Community Commitment Scale with Cross-sectional Survey Validation for Preventing Social Isolation in Older Japanese People

    Directory of Open Access Journals (Sweden)

    Kono Ayumi

    2012-10-01

    Full Text Available Abstract Background Elderly social isolation could be prevented by facilitating communication or mutual helping at the neighborhood level. The helping of elderly neighbors by local volunteers may relate to their community commitment (CC, but ways to measure CC have not been identified. The aim of the present study was to develop a Community Commitment Scale (CCS to measure psychological sense of belonging and socializing in the community among local volunteers, for research in prevention of elderly social isolation. We also tested the CCS in a general population of the elderly. Methods A pilot test of 266 Japanese urban residents was conducted to examine face validity for 24 identified items, of which 12 items were selected for the CCS, based on a 4-point Likert-type scale. The CCS was developed via self-report questionnaires to 859 local volunteers in two suburban cities and to 3484 randomly sampled general residents aged 55 years or older living in one of the cities. To assess concurrent validity, data were collected using the Brief Sense of Community Scale (Peterson; 2008 and two types of single questions on self-efficacy for helping elderly neighbors. Results Item analysis and factor analysis identified 8 items, which were classified between two datasets under the domains of “belonging” and “socializing” in the local volunteers and the general residents. Cronbach’s alpha (which conveyed the internal consistency of the CCS was 0.75 in local volunteers and 0.78 in general residents. The correlation coefficients between the scores of the CCS and BSCS were 0.54 for local volunteers and 0.62 for general residents. ANOVA comparing the CCS between the confidence levels of the two types of single question of self-efficacy on helping elderly neighbors showed a strong relationship in the volunteers and residents. Conclusions These results demonstrate acceptable internal consistency and concurrent validity for the CCS, with the two dimensions

  11. Developing a data acquisition system for the Waste Isolation Pilot Plant's in situ tests

    International Nuclear Information System (INIS)

    McIlmoyle, J.T.; Matalucci, R.V.; Ogden, H.C.

    1986-01-01

    The US Department of Energy is developing the Waste Isolation Pilot Plant in southeastern New Mexico as an R and D facility to demonstrate the safe disposal of radioactive wastes from US defense programs. The in situ tests associated with this facility (659 m below the surface) address technical issues of thermal/structural interaction, waste package performance, and plugging and sealing. Data from these tests are collected by a Data Acquisition System (DAS) consisting of two distinct entities that use the IEEE-488 bus for communication. Up to 5000 data channels can be recorded every 4 hr from extensometers, stressmeters, thermocouples, closure gages, and other special gages. The DAS consists of a multitasking and multiuser ModComp computer housed in a surface trailer that converts the data to engineering units, stores the data in raw and converted form, and provides quick-look plots and data listings. The other part of the DAS consists of the seven underground remote enclosed instrument stations (sheds) located near each test room and up to 1800 m from the surface trailer that contains the instruments and signal conditioning to operate and monitor the tests. The ModComp system manages data acquisition and supports concurrently 12 on-line terminals and two dial-up access lines for data access, gage diagnostics, data handling and updating, and software development. Data are currently acquired from over 4000 gages with 15,000 defined measurands; the system can be expanded to 65,534 measurands. 4 refs., 7 figs

  12. Risk-assessment methodology development for waste isolation in geologic media

    International Nuclear Information System (INIS)

    Stevens, C.A.; Fullwood, R.R.; Amirijafari, B.; Basin, S.L.; Cohen, J.

    1982-12-01

    A review of three documents prepared for the USNRC by Sandia National Laboratories (SNL) is presented. These are NUREG/CR-1634, Volume 4 concerned with the effects of variable hydrology on waste migration; NUREG/CR-2324, a user's manual for SWIFT; and NUREG/2343, a user's manual for DNET. This review completes Task 4 of the detailed technical review of the SNL program for Risk Assessment Methodology Development for Waste Isolation in Geologic Media. In general, these reports exhibit high technical quality that characterizes the SNL work. They are tersely written with little condescension to the non-expert reader for understanding the physical situation being modeled. Indeed, the emphasis is on the mathematical procedures rather than the repository physics, leaving the adequacy of the results presented in many computer plots, pretty much to the interpretation of the reader. Other general comments have been presented previously, such as the data conservatisms, need for data that cannot be measured without disturbing the geometry, and the overall plan for use of the many codes developed in the program

  13. Assessment of effectiveness of Geologic Isolation Systems. The development and application of a geologic simulation model

    International Nuclear Information System (INIS)

    Foley, M.G.; Petrie, G.M.

    1982-03-01

    The Geologic Simulation Model (GSM) developed under the Assessment of Effectiveness of Geologic Isolation Systems (AEGIS) project at the Pacific Northwest Laboratory for the Department of Energy is a quasi-deterministic process-response model which simulates the development of the geologic and hydrologic systems of a ground-water basin for a million years into the future. Effects of natural processes on the ground-water hydrologic system are modeled principally by rate equations. The combined effects and synergistic interactions of different processes are approximated by linear superposition of their effects during discrete time intervals in a stepwise-integration approach. The completed AEGIS GSM was used to generate 500 Monte Carlo simulations of the behavior of the geologic/hydrologic system affecting a hypothetical repository in the Pasco Basin over the next million years. These simulations used data which were not subjected to a review adequate to the needs of a real site performance assessment. However, the general care used in generating the data, and the overall behavior of the GSM suggest that the results are plausible at this time

  14. Comparison of Enzymatic Method Rapid Yeast Plus System with RFLP-PCR for Identification of Isolated Yeast from Vulvovaginal Candidiasis.

    Science.gov (United States)

    Hossein, Moallaei; Mirhendi, Seied Hossein; Brandão, João; Mirdashti, Reza; Rosado, Laura

    2011-09-01

    To compare two identification methods, i.e., restriction fragment length polymorphism (RFLP)-PCR analysis and enzymatic method Rapid TM Yeast Plus System to identify different species causing vulvovaginal candidiasis (VVC). Vaginal discharges of women who had attended the gynecology outpatient clinic of Mobini Hospital in Sabzevar, Iran were collected using cotton swabs and were cultured on Sabouraud dextrose agar. Isolated yeasts were identified by germ-tube testing and Rapid TM Yeast Plus System (Remel USA). For molecular identification, the isolated DNA was amplified with ITS1 and ITS4 universal primers and PCR products digested with the enzyme HpaІІ followed by agarose gel electrophoresis. Epidemiological and clinical features of women with respect to identified species were also evaluated. Out of 231 subjects enrolled, 62 VVC cases were detected. The isolated species were identified as follows: Candida albicans, 24 (38.7%), C. glabrata, 15 (24.2%), C. kefyr, 13 (21.0%) C. krusei, 9 (14.5%), and Saccharomyces cerevisiae, 1 (1.6%) by RFLP-PCR method; whereas findings by Rapid TM Yeast Plus System were C. albicans, 24 (38.7%), C. glabrata, 5 (8%), C. kefyr, 11 (17.7%) C. krusei, 2 (3.2%), S. cerevisiae, 9 (14.5%), and C. tropicalis, 6 (9.6%) as well as other nonpathogenic yeasts, 4 (6.9%). Statistical comparison showed that there is no significant difference in identification of C. albicans by the two methods; although, in this study, it was not true about other species of yeasts. A correlation between clinical and laboratory findings is important as it enables us to administer an appropriate treatment on time.

  15. Comparative method of protein expression and isolation of EBV epitope in E.coli DH5α

    Science.gov (United States)

    Anyndita, Nadya V. M.; Dluha, Nurul; Himmah, Karimatul; Rifa'i, Muhaimin; Widodo

    2017-11-01

    Epstein-Barr Virus (EBV) or human herpes virus 4 (HHV-4) is a virus that infects human B cell and leads to nasopharyngeal carcinoma (NPC). The prevention of this disease remains unsuccessful since the vaccine has not been discovered. The objective of this study is to over-produce EBV gp350/220 epitope using several methods in E.coli DH5α. EBV epitope sequences were inserted into pMAL-p5x vector, then transformed into DH5α E.coli and over-produced using 0.3, 1 and 2 mM IPTG. Plasmid transformation was validated using AflIII restriction enzyme in 0.8% agarose. Periplasmic protein was isolated using 2 comparative methods and then analyzed using SDS-PAGE. Method A produced a protein band around 50 kDa and appeared only at transformant. Method B failed to isolate the protein, indicated by no protein band appearing. In addition, any variations in IPTG concentration didn't give a different result. Thus it can be concluded that even the lowest IPTG concentration is able to induce protein expression.

  16. Evaluation of different phenotypic methods for detection of amp c beta-lactamase producing bacteria in clinical isolates

    International Nuclear Information System (INIS)

    Hassan, A.; Usman, J.; Kalim, F.; Gill, M.M.; Khalid, A.; Iqbal, M.; Ingram, P.

    2013-01-01

    To compare the sensitivity and specificity of different phenotypic methods for detection of Amp C betalactamase producing bacteria. Study Design: Analytical study. Place and Duration of Study: Department of Microbiology, Army Medical College / National University of Sciences and Technology (NUST), Islamabad, Pakistan, from June 2010 to December 2010. Methodology: A total of 150 clinical isolates were screened for presence of Amp C beta-lactamase by using the cefoxitin disc. The confirmatory methods evaluated were inhibitor based assay (boronic acid), Amp C disc test and Amp C Etest. Three dimensional enzyme extract assay was used as the reference method for determining the sensitivity and specificity. Results: Among the total isolates tested, 62.8% bacteria showed the presence of Amp C beta-lactamase by standard three dimensional enzyme extract assay. Among the three methods compared, boronic acid disk test found out to be highly sensitive (88%) and specific (92%) for the detection of Amp C beta-lactamase producing bacteria. Conclusion: Detection of Amp C production is crucial in order to establish the antibiotic therapy and to attain the favourable clinical outcomes. Implementation of simple tests like boronic acid disk tests in the laboratories will help to alleviate the spread of Amp C beta-lactamase harboring organisms. (author)

  17. ISOLATION AND CHARACTERIZATION OF COML, A TRANSCRIPTION UNIT INVOLVED IN COMPETENCE DEVELOPMENT OF BACILLUS-SUBTILIS

    NARCIS (Netherlands)

    VANSINDEREN, D; WITHOFF, S; BOELS, H; VENEMA, G

    1990-01-01

    Using the transformation-deficient mutant M465, which was previously isolated by means of insertional mutagenesis with plasmid pHV60, a transcription unit comL required for genetic competence of Bacillus subtilis was identified. A chromosomal DNA fragment flanking the inserted pHV60 was isolated and

  18. Development of biocontrol agents from food microbial isolates for controlling post-harvest peach brown rot caused by Monilinia fructicola.

    Science.gov (United States)

    Zhou, Ting; Schneider, Karin E; Li, Xiu-Zhen

    2008-08-15

    An unconventional strategy of screening food microbes for biocontrol activity was used to develop biocontrol agents for controlling post-harvest peach brown rot caused by Monilinia fructicola. Forty-four microbial isolates were first screened for their biocontrol activity on apple fruit. Compared with the pathogen-only check, seven of the 44 isolates reduced brown rot incidence by >50%, including four bacteria: Bacillus sp. C06, Lactobacillus sp. C03-b and Bacillus sp. T03-c, Lactobacillus sp. P02 and three yeasts: Saccharomyces delbrueckii A50, S. cerevisiae YE-5 and S. cerevisiae A41. Eight microbial isolates were selected for testing on peaches by wound co-inoculation with mixtures of individual microbial cultures and conidial suspension of M. fructicola. Only two of them showed significant biocontrol activity after five days of incubation at 22 degrees C. Bacillus sp. C06 suppressed brown rot incidence by 92% and reduced lesion diameter by 88% compared to the pathogen-only check. Bacillus sp.T03-c reduced incidence and lesion diameter by 40% and 62%, respectively. The two isolates were compared with Pseudomonas syringae MA-4, a biocontrol agent for post-harvest peach diseases, by immersing peaches in an aliquot containing individual microbial isolates and the pathogen conidia. Treatments with isolates MA-4, C06 and T03-c significantly controlled brown rot by 91, 100, and 100% respectively. However, only isolates MA-4 and C06 significantly reduced brown rot by 80% and 15%, respectively when bacterial cells alone were applied. On naturally infected peaches, both the bacterial culture and its cell-free filtrate of the isolate C06 significantly controlled peach decay resulting in 77 and 90% reduction, respectively, whereas the treatment using only the bacterial cells generally had no effect. Isolate C06 is a single colony isolate obtained from a mesophilic cheese starter, and has been identified belonging to Bacillus amyloliquefaciens. The results have clearly

  19. Systems and methods for enhancing isolation of high-temperature reactor containments

    Energy Technology Data Exchange (ETDEWEB)

    Peterson, Per F.

    2017-09-26

    A high-temperature containment-isolation system for transferring heat from a nuclear reactor containment to a high-pressure heat exchanger is presented. The system uses a high-temperature, low-volatility liquid coolant such as a molten salt or a liquid metal, where the coolant flow path provides liquid free surfaces a short distance from the containment penetrations for the reactor hot-leg and the cold-leg, where these liquid free surfaces have a cover gas maintained at a nearly constant pressure and thus prevent high-pressures from being transmitted into the reactor containment, and where the reactor vessel is suspended within a reactor cavity with a plurality of refractory insulator blocks disposed between an actively cooled inner cavity liner and the reactor vessel.

  20. Antimicrobial Testing Methods & Procedures Developed by EPA's Microbiology Laboratory

    Science.gov (United States)

    We develop antimicrobial testing methods and standard operating procedures to measure the effectiveness of hard surface disinfectants against a variety of microorganisms. Find methods and procedures for antimicrobial testing.

  1. Development of Reverse-Phase HPLC Method for Simultaneous ...

    African Journals Online (AJOL)

    Erah

    Purpose: To develop a simple, sensitive and rapid reverse phase HPLC method for the simultaneous analysis of metoprolol succinate and hydrochlorothiazide in a solid dosage form. Methods: The .... Extraction was carried out three times with.

  2. developed algorithm for the application of british method of concret

    African Journals Online (AJOL)

    t-iyke

    Most of the methods of concrete mix design developed over the years were geared towards manual approach. ... Key words: Concrete mix design; British method; Manual Approach; Algorithm. ..... Statistics for Science and Engineering.

  3. System Identification Methods for Aircraft Flight Control Development and Validation

    Science.gov (United States)

    1995-10-01

    System-identification methods compose a mathematical model, or series of models, : from measurements of inputs and outputs of dynamic systems. This paper : discusses the use of frequency-domain system-identification methods for the : development and ...

  4. Development and validation of a spectroscopic method for the ...

    African Journals Online (AJOL)

    Development and validation of a spectroscopic method for the simultaneous analysis of ... advanced analytical methods such as high pressure liquid ..... equipment. DECLARATIONS ... high-performance liquid chromatography. J Chromatogr.

  5. A simple, rapid method to isolate salt glands for three-dimensional visualization, fluorescence imaging and cytological studies

    Directory of Open Access Journals (Sweden)

    Lim Tit-Meng

    2010-10-01

    Full Text Available Abstract Background Some plants inhabiting saline environment remove salts via the salt glands embedded in the epidermal tissues. Cytological studies of salt glands will provide valuable information to our understanding of the secretory process. Previous studies on salt gland histology relied mainly on two-dimensional microscopic observations of microtome sections. Optical sectioning properties of confocal laser scanning microscope offer alternative approach for obtaining three-dimensional structural information of salt glands. Difficulty in light penetration through intact leaves and interference from neighbouring leaf cells, however, impede the acquiring of good optical salt gland sections and limit its applications in salt gland imaging. Freeing the glands from adjacent leaf tissues will allow better manipulations for three-dimensional imaging through confocal laser scanning microscopy. Results Here, we present a simple and fast method for the isolation of individual salt glands released from the interference of neighbouring cells. About 100-200 salt glands could be isolated from just one cm2 of Avicennia officinalis leaf within hours and microscopic visualization of isolated salt glands was made possible within a day. Using these isolated glands, confocal laser scanning microscopic techniques could be applied and better resolution salt gland images could be achieved. By making use of their intrinsic fluorescent properties, optical sections of the gland cells could be acquired without the use of fluorescent probes and the corresponding three-dimensional images constructed. Useful cytological information of the salt gland cells could also be obtained through the applications of fluorescent dyes (e.g., LysoTracker® Red, FM®4-64, Texas Red®. Conclusions The study of salt glands directly at the glandular level are made possible with the successful isolation of these specialized structures. Preparation of materials for subsequent microscopic

  6. DEVELOPMENT AND EVALUATION OF A SELECTIVE AND INDICATIVE MEDIUM FOR ISOLATION OF ACTINOBACILLUS-PLEUROPNEUMONIAE FROM TONSILS

    DEFF Research Database (Denmark)

    Jakobsen, Marianne; Nielsen, Jens

    1995-01-01

    In order to isolate ActinobacillIus pleuropneumoniae from mixed bacterial flora a selective and indicative medium was developed. The optimal concentrations of antibiotics were determined for selective chocolate agar (S-TSA) and selective blood agar (S-MBA) using a set of 25 strains of A. pleuropn......In order to isolate ActinobacillIus pleuropneumoniae from mixed bacterial flora a selective and indicative medium was developed. The optimal concentrations of antibiotics were determined for selective chocolate agar (S-TSA) and selective blood agar (S-MBA) using a set of 25 strains of A...

  7. Advanced Aqueous Phase Catalyst Development using Combinatorial Methods, Phase II

    Data.gov (United States)

    National Aeronautics and Space Administration — Combinatorial methods are proposed to develop advanced Aqueous Oxidation Catalysts (AOCs) with the capability to mineralize organic contaminants present in effluents...

  8. A method for the isolation and culture of adult rat retinal pigment epithelial (RPE cells to study retinal diseases

    Directory of Open Access Journals (Sweden)

    Janosch Peter Heller

    2015-11-01

    Full Text Available Diseases such as age-related macular degeneration (AMD affect the retinal pigment epithelium (RPE and lead to the death of the epithelial cells and ultimately blindness. RPE transplantation is currently a major focus of eye research and clinical trials using human stem cell-derived RPE cells are ongoing. However, it remains to be established to which extent the source of RPE cells for transplantation affects their therapeutic efficacy and this needs to be explored in animal models. Autotransplantation of RPE cells has attractions as a therapy, but existing protocols to isolate adult RPE cells from rodents are technically difficult, time-consuming, have a low yield and are not optimized for long-term cell culturing. Here, we report a newly devised protocol which facilitates reliable and simple isolation and culture of RPE cells from adult rats. Incubation of a whole rat eyeball in 20 U/ml papain solution for 50 minutes yielded 4 x 104 viable RPE cells. These cells were hexagonal and pigmented upon culture. Using immunostaining, we demonstrated that the cells expressed RPE cell-specific marker proteins including cytokeratin 18 and RPE65, similar to RPE cells in vivo. Additionally, the cells were able to produce and secrete Bruch’s membrane matrix components similar to in vivo situation. Similarly, the cultured RPE cells adhered to isolated Bruch’s membrane as has previously been reported. Therefore, the protocol described in this article provides an efficient method for the rapid and easy isolation of high quantities of adult rat RPE cells. This provides a reliable platform for studying the therapeutic targets, testing the effects of drugs in a preclinical setup and to perform in vitro and in vivo transplantation experiments to study retinal diseases.

  9. Physiologic capacity of well-developed collaterals in patients with isolated left anterior descending artery disease

    International Nuclear Information System (INIS)

    Sakata, Kazuyuki; Yoshida, Hiroshi; Ono, Norihisa; Ohtani, Seiji; Mori, Noriko; Yokoyama, Shoichi; Hoshino, Tsuneo; Kaburagi, Tsuneo; Kurata, Chinori.

    1992-01-01

    To assess the physiologic significance of well-developed collaterals, 34 patients, with isolated left anterior descending artery disease (LAD) and without overt prior myocardial infarction, underwent cardiac catheterization and exercise thallium-201 emission computed tomography. The patients were divided into 3 groups: 11 patients with 90% stenosis of the proximal LAD and without collaterals (group 1), 11 with 99% stenosis of the proximal LAD, and without collaterals (group 2) and 12 with a total occlusion of the proximal LAD which was completely filled by well-developed collaterals (group 3). On left ventriculography, shortening fractions of the anterior wall were significantly reduced in group 2 as compared to groups 1 and 3 (group 1 vs group 2: p<0.01, group 2 vs group 3: p<0.05), which reflected the lower ejection fraction of group 2 (p<0.01 and p<0.05, respectively). The perfusion defects of the anterior wall on both the initial and the delayed images were severer in groups 2 and 3 than in group 1 (group 1 vs group 2 and group 1 vs group 3 on the initial image: p<0.01, for both, group 1 vs group 2 and group 1 vs group 3 on the delayed image: p<0.05, for both). However, recovery of the perfusion defects from the initial image to the delayed image was better in group 3 than in groups 1 and 2 (group 1 vs group 2 and group 1 vs group 3: p<0.05, for both). Therefore, coronary blood flow through well-developed collaterals was considered to be comparable to the flow through a diseased vessel with 90% stenosis at rest. During maximal exercise, blood flow through well-developed collaterals was considered to be comparable to the flow through a diseased vessel with 99% stenosis, although the blood flow through well-developed collaterals was considered to be better than that through 99% stenosis during the recovery period. These findings suggest that patients with well-developed collaterals must be treated like those with severe stenosis. (author)

  10. Condensed summary of the systems prioritization method as a decision-aiding approach for the Waste Isolation Pilot Plant

    International Nuclear Information System (INIS)

    Boak, D.M.; Prindle, N.H.; Lincoln, R.

    1997-01-01

    In March 1994, the US Department of Energy Carlsbad Area Office (DOE/CAO) implemented a performance based decision-aiding method to assist in programmatic prioritization within the Waste Isolation Pilot Plant (WIPP) project. The prioritization was with respect to 40 CFR Part 191.13(a) and 40 CFR part 268.6. U.S. Environmental Protection Agency (EPA) requirements for long-term isolation of radioactive and hazardous wastes. The Systems Prioritization Method (SPM), was designed by Sandia National Laboratories to: (1) identify programmatic options (activities), their costs and durations; (2) analyze combinations of activities in terms of their predicted contribution to long-term performance of the WIPP disposal system; and (3) analyze cost, duration, and performance tradeoffs. SPM results were the basis for activities recommended to DOE/CAO in May 1995. SPM identified eight activities (less than 15% of the 58 proposed for consideration) predicted to be essential in addressing key regulatory issues. The SPM method proved useful for risk or performance-based prioritization in which options are interdependent and system behavior is nonlinear. 10 refs., 2 figs., 1 tab

  11. Isolation of UV-sensitive mutants of mouse L5178Y cells by a cell suspension spotting method

    International Nuclear Information System (INIS)

    Shiomi, T.; Hieda-Shiomi, N.; Sato, K.

    1982-01-01

    We have isolated 56 UV-sensitive mutant clones from a mouse L51 T/t line of L5178Y cells by a cell suspension spotting method. Five mutants have also been isolated from L51 T/t and L5178Y cells by the method reported by Thompson and coworkers. We divided the mutants into two groups, highly sensitive and moderately sensitive mutants, according to their sensitivity to UV irradiation. Fifty-eight mutants were highly sensitive and three were moderately sensitive to UV. The reconstruction experiments indicate that more than 90% of highly sensitive mutants were recovered by the cell suspension spotting method. Frequencies of recovered mutants highly sensitive to UV increased with increasing dose of mutagens. Recovered mutant frequency reached 10(-2) after treatment with 1.5 micrograms/ml of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) (survival 0.2%). Eight UV-sensitive mutants were divided into four complementation groups. These mutants were 2-6 times more sensitive to UV than parental L51 T/t cells in terms of D37 (dose required to reduce survival to 37%). Four representative UV-sensitive mutants which are classified into different complementation groups were examined for their sensitivity to killing by UV, 4-nitroquinoline-1-oxide (4NQO), mitomycin C (MMC), X-rays, and MNNG. All four classes of mutants were found to be cross-sensitive to UV, 4NQO, and MMC, but not sensitive to X-rays and MNNG

  12. Summary of the systems prioritization method (SPM) as a decision-aiding tool for the Waste Isolation Pilot Plant

    International Nuclear Information System (INIS)

    Boak, D.M.; Prindle, N.H.; Lincoln, R.; Mendenhall, F.; Weiner, R.; Bills, R.A.

    1996-01-01

    In March 1994, the Department of Energy Carlsbad Area Office (DOE/CAO) implemented a performance-based planning method to assist in programmatic prioritization within the Waste Isolation Pilot Plant (WIPP) project with respect to applicable Environmental Protection Agency (EPA) long-term performance requirements stated in 40 CFR 191.13(a) and 40 CFR 268.6. This method, the Systems Prioritization Method (SPM), was designed by Sandia National Laboratories (SNL) to: (1) identify programmatic options (activities) and their costs and durations; (2) analyze potential combinations of activities in terms of predicted contribution to long-term performance; and (3) analyze cost, duration, and performance tradeoffs. SPM results were the basis for recommendations to DOE/CAO in May 1995 for prioritization within the WIPP project. This paper presents a summary of the SPM implementation, key results, and lessons learned

  13. Comparative analysis of five DNA isolation protocols and three drying methods for leaves samples of Nectandra megapotamica (Spreng. Mez

    Directory of Open Access Journals (Sweden)

    Leonardo Severo da Costa

    2016-06-01

    Full Text Available The aim of the study was to establish a DNA isolation protocol Nectandra megapotamica (Spreng. Mez., able to obtain samples of high yield and quality for use in genomic analysis. A commercial kit and four classical methods of DNA extraction were tested, including three cetyltrimethylammonium bromide (CTAB-based and one sodium dodecyl sulfate (SDS-based methods. Three drying methods for leaves samples were also evaluated including drying at room temperature (RT, in an oven at 40ºC (S40, and in a microwave oven (FMO. The DNA solutions obtained from different types of leaves samples using the five protocols were assessed in terms of cost, execution time, and quality and yield of extracted DNA. The commercial kit did not extract DNA with sufficient quantity or quality for successful PCR reactions. Among the classic methods, only the protocols of Dellaporta and of Khanuja yielded DNA extractions for all three types of foliar samples that resulted in successful PCR reactions and subsequent enzyme restriction assays. Based on the evaluated variables, the most appropriate DNA extraction method for Nectandra megapotamica (Spreng. Mez. was that of Dellaporta, regardless of the method used to dry the samples. The selected method has a relatively low cost and total execution time. Moreover, the quality and quantity of DNA extracted using this method was sufficient for DNA sequence amplification using PCR reactions and to get restriction fragments.

  14. Comparison of Molecular Typing Methods Useful for Detecting Clusters of Campylobacter jejuni and C. coli Isolates through Routine Surveillance

    Science.gov (United States)

    Taboada, Eduardo; Grant, Christopher C. R.; Blakeston, Connie; Pollari, Frank; Marshall, Barbara; Rahn, Kris; MacKinnon, Joanne; Daignault, Danielle; Pillai, Dylan; Ng, Lai-King

    2012-01-01

    Campylobacter spp. may be responsible for unreported outbreaks of food-borne disease. The detection of these outbreaks is made more difficult by the fact that appropriate methods for detecting clusters of Campylobacter have not been well defined. We have compared the characteristics of five molecular typing methods on Campylobacter jejuni and C. coli isolates obtained from human and nonhuman sources during sentinel site surveillance during a 3-year period. Comparative genomic fingerprinting (CGF) appears to be one of the optimal methods for the detection of clusters of cases, and it could be supplemented by the sequencing of the flaA gene short variable region (flaA SVR sequence typing), with or without subsequent multilocus sequence typing (MLST). Different methods may be optimal for uncovering different aspects of source attribution. Finally, the use of several different molecular typing or analysis methods for comparing individuals within a population reveals much more about that population than a single method. Similarly, comparing several different typing methods reveals a great deal about differences in how the methods group individuals within the population. PMID:22162562

  15. Development of a gas-generation model for the Waste Isolation Pilot Plant

    International Nuclear Information System (INIS)

    Brush, L.H.; Storz, L.J.; Garner, J.W.

    1993-01-01

    Design-basis transuranic (TRU) waste to be emplaced in the Waste Isolation Pilot Plant (WIPP) in southeastern New Mexico may generate significant quantities of gas, which may affect the performance of the WIPP with respect to regulations for radioactive and/or chemically hazardous waste constituents. We are developing a model to predict gas generation in WIPP disposal rooms during and after filling and sealing. Currently, the model includes: (1) oxic and anoxic corrosion of steels and other Fe-base alloys, including passivation and depassivation; (2) microbial degradation of cellulosics with O 2 , NO 3 - , FeO(OH), SO 4 2- , or CO 2 as the electron acceptor; (3) α radiolysis of brine; (4) consumption of CO 2 and, perhaps, H 2 S by Ca(OH) 2 (in cementitious materials) and CaO (a potential backfill additive). The code simulates these processes and interactions among them by converting reactants (steels, cellulosics, etc.) to gases and other products at experimentally observed or estimated rates and plotting temporal reaction paths in three-dimensional phase diagrams for solids in the Fe-H 2 O-CO 2 -H 2 -H 2 S system

  16. Development and characterization of the kefiran-whey protein isolate-TiO2 nanocomposite films.

    Science.gov (United States)

    Zolfi, Mohsen; Khodaiyan, Faramarz; Mousavi, Mohammad; Hashemi, Maryam

    2014-04-01

    Biodegradable kefiran-whey protein isolate (WPI)-titanium dioxide (TiO2) blend films were developed and characterized as a function of incorporating amount of TiO2 nanoparticles (1, 3 and 5% wt.). Results showed that the water vapor permeability, moisture content, moisture absorption and water solubility decreased by increasing the nano-TiO2 content. Mechanical tests revealed the plasticizing effect of TiO2 nanoparticles on the kefiran-WPI-TiO2 film. Addition of TiO2 nanoparticles to kefiran-WPI films significantly decreased tensile strength and Young's modulus, while increased its elongation at break. Differential scanning calorimetry data indicated that the glass transition temperature significantly changed by adding nano-TiO2. X-ray diffraction analysis also demonstrated that crystal type in kefiran-WPI was not affected by incorporation of TiO2 nanoparticles. A uniform distribution at 1 and 3% wt. loading levels of TiO2 nanoparticles was observed using scanning electron microscopy (SEM) micrographs. Copyright © 2014 Elsevier B.V. All rights reserved.

  17. Development of Colletotrichum gloeosporioides isolated from green pepper in different culture media, temperatures, and light regimes

    Directory of Open Access Journals (Sweden)

    Mello Alexandre Furtado Silveira

    2004-01-01

    Full Text Available Control of anthracnose in green pepper involves the use of resistant varieties and/or fungicides. The selection of varieties and efficient products demands great amounts of conidia as inoculum. It is thus necessary to optimize the production of Colletotrichum gloeosporioides conidia in the laboratory, establishing the best conditions for fungus development. The present study aimed at determining the most favorable culture media, temperature, and light conditions for the production of fungus inoculum. The fungus was isolated from green pepper fruits (Capsicum annuum L. and transferred to four culture media (PDA, oat, filtered pepper extract, and autoclaved pepper extract, under different temperatures (15, 20, 25, 30, and 35ºC and light conditions (24h dark, and 24h light. Colony growth was evaluated after 7 and 12 days of incubation. No differences were found between the culture media. However, the greatest number of conidia was obtained from colonies grown in oat medium at 25ºC. Temperatures of 20 and 25ºC were the most favorable for colony growth and sporulation. Higher sporulation was obtained under incubation in constant light. Cultivation of C. gloeosporioides in oat medium, at 25ºC, and constant light is recommended.

  18. Development of an Immunomagnetic Separation Method for Viable Salmonella Typhimurium Detected by Flow Cytometry

    DEFF Research Database (Denmark)

    Ahmed, Shakil; Rubahn, Horst-Günter; Erdmann, Helmut

    2016-01-01

    for detection of food-related bacteria. In this study, a flow cytometry based immunomagnetic separation (IMS) method for the isolation and enrichment of Salmonella Typhimurium from liquid samples was developed and optimized. Both polyclonal and monoclonal antibodies have been used to couple with 1 micron sized...... and bacteria, immunocapture time, staining and buffering conditions for the viability assays were optimized. The capture efficiency of IMS was>98% for a range of Salmonella Typhimurium cell concentrations from 103 to 105/mL using 108/mL bead concentration. The method proved to have high (98%) specificity...

  19. Isolated Ficus trees deliver dual conservation and development benefits in a rural landscape

    DEFF Research Database (Denmark)

    Cottee-Jones, H. Eden W.; Bajpai, Omesh; Chaudhary, Lal B.

    2015-01-01

    strategy to manage the availability of valuable plant resources, facilitated by the use of isolated Ficus trees as restoration nuclei. We surveyed the plants growing under 207 isolated trees in Assam, India, and categorized them according to their local human-uses. We found that Ficus trees were associated...... densities in most categories. Community management practices that conserve isolated Ficus trees, and restrict livestock grazing and high-intensity land-use in their vicinity, can promote plant growth and the provision of important local resources....

  20. Development of Probiotic Milk Drinks Using Probiotic Strain Isolated From Local Yogurt

    Directory of Open Access Journals (Sweden)

    Md. Ahmadul Islam

    2018-08-01

    Full Text Available Obtained result suggested that, Isolate 1 and Isolate 2 during storage at 5±1oC for 15days ranged between 8.75 – 9.55 log cfu/ml those were higher than 5 log cfu/ml. That means isolated lactic acid bacteria (LAB contain probiotic characteristics & among three samples, the probiotic milk drink that was mixed with 10% mango juice was the most acceptable. [Fundam Appl Agric 2018; 3(2.000: 446-452

  1. Isolation and detection of Listeria monocytogenes in poultry meat by standard culture methods and PCR

    Science.gov (United States)

    Kureljušić, J.; Rokvić, N.; Jezdimirović, N.; Kureljušić, B.; Pisinov, B.; Karabasil, N.

    2017-09-01

    Listeria is the genus of a bacteria found in soil and water and some animals, including poultry and cattle. It can be present in raw milk and food made from raw milk. It can also live in food processing plants and contaminate a variety of processed meats. Microscopically, Listeria species appear as small, Gram-positive rods, which are sometimes arranged in short chains. In direct smears, they can be coccoid, so they can be mistaken for streptococci. Longer cells can resemble corynebacteria. Flagella are produced at room temperature but not at 37°C. Haemolytic activity on blood agar has been used as a marker to distinguish Listeria monocytogenes among other Listeria species, but it is not an absolutely definitive criterion. Further biochemical characterization is necessary to distinguish between the different Listeria species. The objective of this study was to detect, isolate and identify Listeria monocytogenes from poultry meat. Within a period of six months from January to June 2017, a total of 15 samples were collected. Three samples were positive for the presence of Listeria monocytogenes. Biochemical and microbiological tests as well as PCR technique using specific primers were used to confirm L. Monocytogenes in the samples.

  2. Control Method Based on Demand Response Needs of Isolated Bus Regulation with Series-Resonant Converters for Residential Photovoltaic Systems

    Directory of Open Access Journals (Sweden)

    Shu-Huai Zhang

    2017-05-01

    Full Text Available Considering the effects of isolation and high efficiency, a series-resonant DC-DC converter (L-L-C type, with two inductors and a capacitor has been introduced into a residential photovoltaic (PV generation and storage system in this work, and a voltage gain curve upwarp drifting problem was found. In this paper, the reason of upwarp drifting in the voltage gain curve is given, and a new changing topological control method to solve the voltage regulation problem under light load conditions is proposed. Firstly, the ideal and actual first harmonic approximation (FHA models are given, and this drifting problem is ascribed to the multiple peaks of higher-order resonance between resonant tank and parasitic capacitors. Then the paper presents the pulse-frequency-modulation (PFM driver signals control method to translate the full-bridge LLC into a half-bridge LLC converter, and with this method the voltage gain could easily be reduced by half. Based on this method, the whole voltage and resonant current sharing control methods in on-line and off-line mode are proposed. The parameters design and optimization methods are also discussed in detail. Finally, a residential PV system platform based on the proposed parallel 7-kW full-bridge LLC converter is built to verify the proposed control method and theoretical analysis.

  3. Comparison of culture based methods for the isolation of Clostridium difficile from stool samples in a research setting.

    Science.gov (United States)

    Lister, Michelle; Stevenson, Emma; Heeg, Daniela; Minton, Nigel P; Kuehne, Sarah A

    2014-08-01

    Effective isolation of Clostridium difficile from stool samples is important in the research setting, especially where low numbers of spores/vegetative cells may be present within a sample. In this study, three protocols for stool culture were investigated to find a sensitive, cost effective and timely method of C. difficile isolation. For the initial enrichment step, the effectiveness of two different rich media, cycloserine-cefoxitin fructose broth (CCFB) and cycloserine-cefoxitin mannitol broth with taurocholate and lysozyme (CCMB-TAL) were compared. For the comparison of four different, selective solid media; Cycloserine-cefoxitin fructose agar (CCFA), Cycloserine-cefoxitin egg yolk agar (CCEY), ChromID C. difficile and tryptone soy agar (TSA) with 5% sheep's blood with and without preceding broth enrichment were used. As a means to enable differentiation between C. difficile and other fecal flora, the effectiveness of the inclusion of a pH indictor (1% Neutral Red), was also evaluated. The data derived indicated that CCFB is more sensitive than CCMB-TAL, however, the latter had an improved recovery rate. A broth enrichment step had a reduced sensitivity over direct plating. ChromID C. difficile showed the best recovery rate whereas CCEY egg yolk agar was the most sensitive of the four. The addition of 1% Neutral Red did not show sufficient colour change when added to CCEY egg yolk agar to be used as a differential medium. For a low cost, timely and sensitive method of isolating C. difficile from stool samples we recommend direct plating onto CCEY egg yolk agar after heat shock. Copyright © 2014 The Authors. Published by Elsevier Ltd.. All rights reserved.

  4. Generating and testing methods for consumer-oriented product development

    International Nuclear Information System (INIS)

    2001-10-01

    In order to obtain a good insight into various design methods that can be used by product developers to enable them to develop and test useful domotics products (domotics: intelligent systems for the home), an inventory has been made of the methods used in the Netherlands. The inventory is directed at two categories of methods: (1) Methods of getting better acquainted with the user and/or the problem, and of generating novel solutions: generative methods; and (2) Methods of assessing solutions (through various phases of the designing process): testing methods. The first category of methods concentrates on the designing process. In other words: how can the designer realise as much as possible of the workability of (domotics) products during the designing process? The second category aims at testing a design (in whatever shape: drawing, prototype, functional computer animation, etc.) through its users. These are methods of assessing a design at various stages of the designing process [nl

  5. Comparison of pathogen DNA isolation methods from large volumes of whole blood to improve molecular diagnosis of bloodstream infections.

    Directory of Open Access Journals (Sweden)

    Anne J M Loonen

    Full Text Available For patients suffering from bloodstream infections (BSI molecular diagnostics from whole blood holds promise to provide fast and adequate treatment. However, this approach is hampered by the need of large blood volumes. Three methods for pathogen DNA isolation from whole blood were compared, i.e. an enzymatic method (MolYsis, 1-5 ml, the novel non-enzymatic procedure (Polaris, 1-5 ml, and a method that does not entail removal of human DNA (Triton-Tris-EDTA EasyMAG, 200 µl. These methods were evaluated by processing blood spiked with 0-1000 CFU/ml of Staphylococcus aureus, Pseudomonas aeruginosa and Candida albicans. Downstream detection was performed with real-time PCR assays. Polaris and MolYsis processing followed by real-time PCRs enabled pathogen detection at clinically relevant concentrations of 1-10 CFU/ml blood. By increasing sample volumes, concurrent lower cycle threshold (Ct values were obtained at clinically relevant pathogen concentrations, demonstrating the benefit of using larger blood volumes. A 100% detection rate at a concentration of 10 CFU/ml for all tested pathogens was obtained with the Polaris enrichment, whereas comparatively lower detection rates were measured for MolYsis (50-67% and EasyMAG (58-79%. For the samples with a concentration of 1 CFU/ml Polaris resulted in most optimal detection rates of 70-75% (MolYsis 17-50% and TTE-EasyMAG 20-36%. The Polaris method was more reproducible, less labour intensive, and faster (45 minutes (including Qiagen DNA extraction vs. 2 hours (MolYsis. In conclusion, Polaris and MolYsis enrichment followed by DNA isolation and real-time PCR enables reliable and sensitive detection of bacteria and fungi from 5 ml blood. With Polaris results are available within 3 hours, showing potential for improved BSI diagnostics.

  6. Development of Early Warning Methods for Electric Power Systems

    DEFF Research Database (Denmark)

    Jóhannsson, Hjörtur

    This thesis concerns the development of methods that can provide, in realtime, an early warning for an emerging blackout in electric power systems. The blackout in E-Denmark and S-Sweden on September 23, 2003 is the main motivation for the method development. The blackout was caused by occurrence...

  7. How Qualitative Methods Can be Used to Inform Model Development.

    Science.gov (United States)

    Husbands, Samantha; Jowett, Susan; Barton, Pelham; Coast, Joanna

    2017-06-01

    Decision-analytic models play a key role in informing healthcare resource allocation decisions. However, there are ongoing concerns with the credibility of models. Modelling methods guidance can encourage good practice within model development, but its value is dependent on its ability to address the areas that modellers find most challenging. Further, it is important that modelling methods and related guidance are continually updated in light of any new approaches that could potentially enhance model credibility. The objective of this article was to highlight the ways in which qualitative methods have been used and recommended to inform decision-analytic model development and enhance modelling practices. With reference to the literature, the article discusses two key ways in which qualitative methods can be, and have been, applied. The first approach involves using qualitative methods to understand and inform general and future processes of model development, and the second, using qualitative techniques to directly inform the development of individual models. The literature suggests that qualitative methods can improve the validity and credibility of modelling processes by providing a means to understand existing modelling approaches that identifies where problems are occurring and further guidance is needed. It can also be applied within model development to facilitate the input of experts to structural development. We recommend that current and future model development would benefit from the greater integration of qualitative methods, specifically by studying 'real' modelling processes, and by developing recommendations around how qualitative methods can be adopted within everyday modelling practice.

  8. Development and Validation of Analytical Method for Losartan ...

    African Journals Online (AJOL)

    Development and Validation of Analytical Method for Losartan-Copper Complex Using UV-Vis Spectrophotometry. ... Tropical Journal of Pharmaceutical Research ... Purpose: To develop a new spectrophotometric method for the analysis of losartan potassium in pharmaceutical formulations by making its complex with ...

  9. Developing a reliable signal wire attachment method for rail.

    Science.gov (United States)

    2014-11-01

    The goal of this project was to develop a better attachment method for rail signal wires to improve the reliability of signaling : systems. EWI conducted basic research into the failure mode of current attachment methods and developed and tested a ne...

  10. Rural Latinos' mental wellbeing: a mixed-methods pilot study of family, environment and social isolation factors.

    Science.gov (United States)

    Stacciarini, Jeanne-Marie R; Smith, Rebekah; Garvan, Cynthia Wilson; Wiens, Brenda; Cottler, Linda B

    2015-05-01

    Upon immigration to the rural areas in the US, Latino families may experience cultural, geographic, linguistic and social isolation, which can detrimentally affect their wellbeing by acting as chronic stressors. Using a community engagement approach, this is a pilot mixed-method study with an embedded design using concurrent qualitative and quantitative data. The purpose of this study is to evaluate family and social environments in terms of protective factors and modifiable risks associated with mental well-being in Latino immigrants living in rural areas of Florida. Latino immigrant mother and adolescent dyads were interviewed by using in-depth ethnographic semistructured interviews and subsequent quantitative assessments, including a demographic questionnaire and three structured instruments: the Family Environment Scale Real Form, the SF-12v2™ Health Survey and the short version (eight items) of PROMIS Health Organization Social Isolation. This mixed-method pilot study highlighted how family, rural, and social environments can protect or impair wellbeing in rural Latino immigrant mother and adolescent dyads.

  11. Method Development and Monitoring of Cyanotoxins in Water

    Science.gov (United States)

    This presentation describes method development of two ambient water LC/MS/MS methods for microcystins, cylindrospermopsin and anatoxin-a. Ruggedness of the methods will be demonstrated by evaluation of quality control samples derived from various water bodies across the country.

  12. Development of a Research Methods and Statistics Concept Inventory

    Science.gov (United States)

    Veilleux, Jennifer C.; Chapman, Kate M.

    2017-01-01

    Research methods and statistics are core courses in the undergraduate psychology major. To assess learning outcomes, it would be useful to have a measure that assesses research methods and statistical literacy beyond course grades. In two studies, we developed and provided initial validation results for a research methods and statistical knowledge…

  13. Comparison of two silica-based extraction methods for DNA isolation from bones.

    Science.gov (United States)

    Rothe, Jessica; Nagy, Marion

    2016-09-01

    One of the most demanding DNA extractions is from bones and teeth due to the robustness of the material and the relatively low DNA content. The greatest challenge is due to the manifold nature of the material, which is defined by various factors, including age, storage, environmental conditions, and contamination with inhibitors. However, most published protocols do not distinguish between different types or qualities of bone material, but are described as being generally applicable. Our laboratory works with two different extraction methods based on silica membranes or the use of silica beads. We compared the amplification success of the two methods from bone samples with different qualities and in the presence of inhibitors. We found that the DNA extraction using the silica membrane method results an in higher DNA yield but also in a higher risk of co-extracting impurities, which can act as inhibitors. In contrast the silica beads method shows decreased co-extraction of inhibitors but also less DNA yield. Related to our own experiences it has to be considered that each bone material should be reviewed independently regarding the analysis and extraction method. Therefore, the most ambitious task is determining the quality of the bone material, which requires substantial experience. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  14. Alu polymerase chain reaction: A method for rapid isolation of human-specific sequences from complex DNA sources

    International Nuclear Information System (INIS)

    Nelson, D.L.; Ledbetter, S.A.; Corbo, L.; Victoria, M.F.; Ramirez-Solis, R.; Webster, T.D.; Ledbetter, D.H.; Caskey, C.T.

    1989-01-01

    Current efforts to map the human genome are focused on individual chromosomes or smaller regions and frequently rely on the use of somatic cell hybrids. The authors report the application of the polymerase chain reaction to direct amplification of human DNA from hybrid cells containing regions of the human genome in rodent cell backgrounds using primers directed to the human Alu repeat element. They demonstrate Alu-directed amplification of a fragment of the human HPRT gene from both hybrid cell and cloned DNA and identify through sequence analysis the Alu repeats involved in this amplification. They also demonstrate the application of this technique to identify the chromosomal locations of large fragments of the human X chromosome cloned in a yeast artificial chromosome and the general applicability of the method to the preparation of DNA probes from cloned human sequences. The technique allows rapid gene mapping and provides a simple method for the isolation and analysis of specific chromosomal regions

  15. Method of isolation of traces of americium by using the +6 oxidation state properties

    International Nuclear Information System (INIS)

    Kwinta, Jean; Michel, Jean-Jacques

    1969-05-01

    The authors present a method to separate traces of americium from a solution containing fission products and actinides. This method comprises the following steps: firstly, the oxidation of americium at the +6 state by ammonium persulfate and carrying over of actinides and III and IV lanthanides by lanthanum fluoride; secondly, the reduction by hydrazine of the oxidized americium and carrying over of the reduced americium by lutetium fluoride; and thirdly, the americium-lutetium separation by selective extractions either with di 2 ethyl hexyl phosphoric acid, or by fractionated elution on an anionic resin column by a mixture of nitric acid and methanol [fr

  16. A novel method for isolation of histones from serum and its implications in therapeutics and prognosis of solid tumours.

    Science.gov (United States)

    Reddy, Divya; Khade, Bharat; Pandya, Riddhi; Gupta, Sanjay

    2017-01-01

    Dysregulation in post-translational modifications of histones and their modifiers are now well-recognized as a hallmark of cancer and can be used as biomarkers and potential therapeutic targets for disease progression and prognosis. In most solid tumours, a biopsy is challenging, costly, painful or potentially risky for the patient. Therefore, non-invasive methods like 'liquid biopsy' for analysis of histone modifications and their modifiers if possible will be helpful in the better clinical management of cancer patients. Here, we have developed a cost-effective and time-efficient protocol for isolation of circulating histones from serum of solid tumor, HCC, called Dual Acid Extraction (DAE) protocol and have confirmed by mass spectrometry. Also, we measured the activity of HDACs and HATs in serum samples. The serum purified histones were profiled for changes in histone PTMs and have shown a comparable pattern of modifications like acetylation (H4K16Ac), methylation (H4K20Me3, H3K27Me3, H3K9Me3) and phosphorylation (γ-H2AX and H3S10P) to paired cancer tissues. Profiling for the histone PTM changes in various other organs of normal and tumor bearing animal suggests that the changes in the histone PTMs observed in the tumor serum is indeed due to changes in the tumor tissue only. Further, we demonstrate that the observed hypo-acetylation of histone H4 in tissue and serum samples of tumor bearing animals corroborated with the elevated HDAC activity in both samples compared to normal. Interestingly, human normal and tumor serum samples also showed elevated HDAC activity with no significant changes in HAT activity. Our study provides the first evidence in the context of histone PTMs and modifiers that liquid biopsy is a valuable predictive tool for monitoring disease progression. Importantly, with the advent of drugs that target specific enzymes involved in the epigenetic regulation of gene expression, liquid biopsy-based 'real time' monitoring will be useful for

  17. An alternative method to isolate protease and phospholipase A2 toxins from snake venoms based on partitioning of aqueous two-phase systems

    Directory of Open Access Journals (Sweden)

    GN Gómez

    2012-01-01

    Full Text Available Snake venoms are rich sources of active proteins that have been employed in the diagnosis and treatment of health disorders and antivenom therapy. Developing countries demand fast economical downstream processes for the purification of this biomolecule type without requiring sophisticated equipment. We developed an alternative, simple and easy to scale-up method, able to purify simultaneously protease and phospholipase A2 toxins from Bothrops alternatus venom. It comprises a multiple-step partition procedure with polyethylene-glycol/phosphate aqueous two-phase systems followed by a gel filtration chromatographic step. Two single bands in SDS-polyacrylamide gel electrophoresis and increased proteolytic and phospholipase A2 specific activities evidence the homogeneity of the isolated proteins.

  18. Leaf cuticles as mediators of environmental influences: new developments in the use of isolated cuticles

    International Nuclear Information System (INIS)

    Scherbatskoy, T.

    1994-01-01

    Isolated leaf cuticles have been used in our research to characterize trans-cuticular ion diffusion rates under various environmental treatments and to measure cuticular attenuation of solar radiation. These studies have been conducted to understand better the role of the leaf cuticle as a protective barrier against potential environmental stressors including acid rain, ozone and ultraviolet radiation. These studies provide examples of a variety of current research uses for isolated leaf cuticles. Ion permeability coefficients and exchange rates in isolated cuticles of Acer, Prunus and Citrus species have been studied using several experimental approaches, including measurement of adsorption kinetics, electrical potential and conductance, and perfusion rates. Measured electrical (diffusion) potentials under KCl gradients across isolated cuticles are positive, indicating greater cation permeability. Electrical potentials (and permeability coefficients) vary with ionic strength and pH, and affect the driving force for ion diffusion through cuticles. Ion permeability in cuticles of Prunus serotina foliage was affected by experimental exposure to ozone and ultraviolet treatments. These studies indicate that cuticle permeability properties can be significantly altered by environmental factors. These and related studies on the ion exchange kinetics of Acer saccharum leaf cuticles suggest that foliar ''leaching'' is dominated by cuticle surface exchange mechanisms, with the magnitude of cuticular ion permeation being relatively small. In working with various hardwood species, we observed that the success rate for cuticle isolation varies with tree species and time of year. Scanning electron micrographs of inner cuticle surfaces indicate that the effects of enzymatic digestion could vary with exposure time, possibly affecting transport properties of isolated cuticles. Experimental work to test this in Citrus, however, showed no significant effect of isolation time on

  19. Segmentation of isolated MR images: development and comparison of neuronal networks

    International Nuclear Information System (INIS)

    Paredes, R.; Robles, M.; Marti-Bonmati, L.; Masia, L.

    1998-01-01

    Segmentation defines the capacity to differentiate among types of tissues. In MR. it is frequently applied to volumetric determinations. Digital images can be segmented in a number of ways; neuronal networks (NN) can be employed for this purpose. Our objective was to develop algorithms for automatic segmentation using NN and apply them to central nervous system MR images. The segmentation obtained with NN was compared with that resulting from other procedures (region-growing and K means). Each NN consisted of two layers: one based on unsupervised training, which was utilized for image segmentation in sets of K, and a second layer associating each set obtained by the preceding layer with the real set corresponding to the previously segmented objective image. This NN was trained with previously segmented images with supervised regions-growing algorithms and automatic K means. Thus, 4 different segmentation were obtained: region-growing, K means, NN with region-growing and NN with K means. The tissue volumes corresponding to cerebrospinal fluid, gray matter and white matter obtained with the 4 techniques were compared and the most representative segmented image was selected qualitatively by averaging the visual perception of 3 radiologists. The segmentation that best corresponded to the visual perception of the radiologists was that consisting of trained NN with region-growing. In comparison, the other 3 algorithms presented low percentage differences (mean, 3.44%). The mean percentage error for the 3 tissues from these algorithms was lower for region-growing segmentation (2.34%) than for trained NN with K means (3.31%) and for automatic K-means segmentation (4.66%). Thus, NN are reliable in the automation of isolated MR image segmentation. (Author) 12 refs

  20. Enhanced computational methods for quantifying the effect of geographic and environmental isolation on genetic differentiation

    NARCIS (Netherlands)

    Botta, Filippo; Eriksen, Casper; Fontaine, Michael Christophe; Guillot, Gilles

    2015-01-01

    In a recent paper, Bradburd et al. (2013) proposed a model to quantify the relative effect ofgeographic and environmental distance on genetic differentiation. Here, we enhance this method in several ways. 1. We modify the covariance model so as to fit better with mainstream geostatistical models and

  1. Long-term climate change assessment study plan for the Hanford Site Permanent Isolation Barrier Development Program

    International Nuclear Information System (INIS)

    Petersen, K.L.; Chatters, J.C.; Waugh, W.J.

    1993-05-01

    The Hanford Site Permanent Isolation Barrier Development Program (Barrier Development Program) was organized to develop the technology needed to provide an in-place disposal capability for low-level nuclear waste for the US Department of Energy at the Hanford Site in south-central Washington. The goal of the Barrier Development Program is to provide defensible evidence that final barrier design(s) will adequately control water infiltration, plant and animal intrusion, and wind and water erosion for a minimum of 1,000 yr; to isolate wastes from the accessible environment; and to use markers to warn inadvertent human intruders. Evidence for barrier performance will be obtained by conducting laboratory experiments, field tests, computer modeling, and other studies that establish confidence in the barrier's ability to meet its 1,000-yr design life

  2. Social Isolation During Postweaning Development Causes Hypoactivity of Neurons in the Medial Nucleus of the Male Rat Amygdala

    Science.gov (United States)

    Adams, Thomas; Rosenkranz, J Amiel

    2016-01-01

    Children exposed to neglect or social deprivation are at heightened risk for psychiatric disorders and abnormal social patterns as adults. There is also evidence that prepubertal neglect in children causes abnormal metabolic activity in several brain regions, including the amygdala area. The medial nucleus of the amygdala (MeA) is a key region for performance of social behaviors and still undergoes maturation during the periadolescent period. As such, the normal development of this region may be disrupted by social deprivation. In rodents, postweaning social isolation causes a range of deficits in sexual and agonistic behaviors that normally rely on the posterior MeA (MeAp). However, little is known about the effects of social isolation on the function of MeA neurons. In this study, we tested whether postweaning social isolation caused abnormal activity of MeA neurons. We found that postweaning social isolation caused a decrease of in vivo firing activity of MeAp neurons, and reduced drive from excitatory afferents. In vitro electrophysiological studies found that postweaning social isolation caused a presynaptic impairment of excitatory input to the dorsal MeAp, but a progressive postsynaptic reduction of membrane excitability in the ventral MeAp. These results demonstrate discrete, subnucleus-specific effects of social deprivation on the physiology of MeAp neurons. This pathophysiology may contribute to the disruption of social behavior after developmental social deprivation, and may be a novel target to facilitate the treatment of social disorders. PMID:26677945

  3. Social Isolation During Postweaning Development Causes Hypoactivity of Neurons in the Medial Nucleus of the Male Rat Amygdala.

    Science.gov (United States)

    Adams, Thomas; Rosenkranz, J Amiel

    2016-06-01

    Children exposed to neglect or social deprivation are at heightened risk for psychiatric disorders and abnormal social patterns as adults. There is also evidence that prepubertal neglect in children causes abnormal metabolic activity in several brain regions, including the amygdala area. The medial nucleus of the amygdala (MeA) is a key region for performance of social behaviors and still undergoes maturation during the periadolescent period. As such, the normal development of this region may be disrupted by social deprivation. In rodents, postweaning social isolation causes a range of deficits in sexual and agonistic behaviors that normally rely on the posterior MeA (MeAp). However, little is known about the effects of social isolation on the function of MeA neurons. In this study, we tested whether postweaning social isolation caused abnormal activity of MeA neurons. We found that postweaning social isolation caused a decrease of in vivo firing activity of MeAp neurons, and reduced drive from excitatory afferents. In vitro electrophysiological studies found that postweaning social isolation caused a presynaptic impairment of excitatory input to the dorsal MeAp, but a progressive postsynaptic reduction of membrane excitability in the ventral MeAp. These results demonstrate discrete, subnucleus-specific effects of social deprivation on the physiology of MeAp neurons. This pathophysiology may contribute to the disruption of social behavior after developmental social deprivation, and may be a novel target to facilitate the treatment of social disorders.

  4. Verification of analysis methods for predicting the behaviour of seismically isolated nuclear structures. Final report of a co-ordinated research project 1996-1999

    International Nuclear Information System (INIS)

    2002-06-01

    This report is a summary of the work performed under a co-ordinated research project (CRP) entitled Verification of Analysis Methods for Predicting the Behaviour of Seismically isolated Nuclear Structures. The project was organized by the IAEA on the recommendation of the IAEA's Technical Working Group on Fast Reactors (TWGFR) and carried out from 1996 to 1999. One of the primary requirements for nuclear power plants and facilities is to ensure safety and the absence of damage under strong external dynamic loading from, for example, earthquakes. The designs of liquid metal cooled fast reactors (LMFRs) include systems which operate at low pressure and include components which are thin-walled and flexible. These systems and components could be considerably affected by earthquakes in seismic zones. Therefore, the IAEA through its advanced reactor technology development programme supports the activities of Member States to apply seismic isolation technology to LMFRs. The application of this technology to LMFRs and other nuclear plants and related facilities would offer the advantage that standard designs may be safely used in areas with a seismic risk. The technology may also provide a means of seismically upgrading nuclear facilities. Design analyses applied to such critical structures need to be firmly established, and the CRP provided a valuable tool in assessing their reliability. Ten organizations from India, Italy, Japan, the Republic of Korea, the Russian Federation, the United Kingdom, the United States of America and the European Commission co-operated in this CRP. This report documents the CRP activities, provides the main results and recommendations and includes the work carried out by the research groups at the participating institutes within the CRP on verification of their analysis methods for predicting the behaviour of seismically isolated nuclear structures

  5. Easy xeno-free and feeder-free method for isolating and growing limbal stromal and epithelial stem cells of the human cornea.

    Directory of Open Access Journals (Sweden)

    Djida Ghoubay-Benallaoua

    Full Text Available Epithelial and stromal stem cells are required to maintain corneal transparency. The aim of the study was to develop a new method to isolate and grow both corneal stromal (SSC and epithelial limbal (LSC stem cells from small human limbal biopsies under culture conditions in accordance with safety requirements mandatory for clinical use in humans. Superficial limbal explants were retrieved from human donor corneo-scleral rims. Human limbal cells were dissociated by digestion with collagenase A, either after epithelial scraping or with no scraping. Isolated cells were cultured with Essential 8 medium (E8, E8 supplemented with EGF (E8+ or Green's medium with 3T3 feeder-layers. Cells were characterized by immunostaining, RT-qPCR, colony forming efficiency, sphere formation, population doubling, second harmonic generation microscopy and differentiation potentials. LSC were obtained from unscraped explants in E8, E8+ and Green's media and were characterized by colony formation and expression of PAX6, ΔNP63α, Bmi1, ABCG2, SOX9, CK14, CK15 and vimentin, with a few cells positive for CK3. LSC underwent 28 population doublings still forming colonies. SSC were obtained from both scraped and unscraped explants in E8 and E8+ media and were characterized by sphere formation, expression of PAX6, SOX2, BMI1, NESTIN, ABCG2, KERATOCAN, VIMENTIN, SOX9, SOX10 and HNK1, production of collagen fibrils and differentiation into keratocytes, fibroblasts, myofibroblasts, neurons, adipocytes, chondrocytes and osteocytes. SSC underwent 48 population doublings still forming spheres, Thus, this new method allows both SSC and LSC to be isolated from small superficial limbal biopsies and to be primary cultured in feeder-free and xeno-free conditions, which will be useful for clinical purposes.

  6. Modeling of Self-Excited Isolated Permanent Magnet Induction Generator Using Iterative Numerical Method

    Directory of Open Access Journals (Sweden)

    Mohamed Mostafa R.

    2016-01-01

    Full Text Available Self-Excited Permanent Magnet Induction Generator (PMIG is commonly used in wind energy generation systems. The difficulty of Self-Excited Permanent Magnet Induction Generator (SEPMIG modeling is the circuit parameters of the generator vary at each load conditions due to the a change in the frequency and stator voltage. The paper introduces a new modeling for SEPMIG using Gauss-sidle relaxation method. The SEPMIG characteristics using the proposed method are studied at different load conditions according to the wind speed variation, load impedance changes and different shunted capacitor values. The system modeling is investigated due to the magnetizing current variation, the efficiency variation, the power variation and power factor variation. The proposed modeling system satisfies high degree of simplicity and accuracy.

  7. Method of targeted delivery of laser beam to isolated retinal rods by fiber optics.

    Science.gov (United States)

    Sim, Nigel; Bessarab, Dmitri; Jones, C Michael; Krivitsky, Leonid

    2011-11-01

    A method of controllable light delivery to retinal rod cells using an optical fiber is described. Photo-induced current of the living rod cells was measured with the suction electrode technique. The approach was tested with measurements relating the spatial distribution of the light intensity to photo-induced current. In addition, the ion current responses of rod cells to polarized light at two different orientation geometries of the cells were studied.

  8. Automated segmentation and isolation of touching cell nuclei in cytopathology smear images of pleural effusion using distance transform watershed method

    Science.gov (United States)

    Win, Khin Yadanar; Choomchuay, Somsak; Hamamoto, Kazuhiko

    2017-06-01

    The automated segmentation of cell nuclei is an essential stage in the quantitative image analysis of cell nuclei extracted from smear cytology images of pleural fluid. Cell nuclei can indicate cancer as the characteristics of cell nuclei are associated with cells proliferation and malignancy in term of size, shape and the stained color. Nevertheless, automatic nuclei segmentation has remained challenging due to the artifacts caused by slide preparation, nuclei heterogeneity such as the poor contrast, inconsistent stained color, the cells variation, and cells overlapping. In this paper, we proposed a watershed-based method that is capable to segment the nuclei of the variety of cells from cytology pleural fluid smear images. Firstly, the original image is preprocessed by converting into the grayscale image and enhancing by adjusting and equalizing the intensity using histogram equalization. Next, the cell nuclei are segmented using OTSU thresholding as the binary image. The undesirable artifacts are eliminated using morphological operations. Finally, the distance transform based watershed method is applied to isolate the touching and overlapping cell nuclei. The proposed method is tested with 25 Papanicolaou (Pap) stained pleural fluid images. The accuracy of our proposed method is 92%. The method is relatively simple, and the results are very promising.

  9. Long-term climate change assessment study plan for the Hanford Site permanent isolation Barrier Development Program

    International Nuclear Information System (INIS)

    Petersen, K.L.; Chatters, J.C.; Waugh, W.J.

    1992-07-01

    The Hanford Site Permanent Isolation Barrier Development Program (Barrier Development Program) was organized to develop the technology needed to provide an in-place disposal capability for the Hanford Site (Adams and Wing 1986; Wig and Gee 1990). The goal of the Barrier Development Program is to provide defensible evidence that final barrier design(s) will adequately control water infiltration; plant and animal intrusion; and wind and water erosion for a minimum of 1,000 yr and isolate wastes from the accessible environment and warm inadvertent human intruders using markers. This document describes the long-term climate change studies planned to support the Barrier Development Program. The plan outlines a multi-year and multi-discipline approach to assess long-term climate change issues and to help optimize the design of the permanent isolation barriers. A multi-disciplinary approach to climatic data acquisition will be responsible for obtaining needed information for concurrent barrier tasks and for developing a local climate forecast model. This model will couple past climate patterns with models of regional and global climate drivers to provide bounding conditions for barrier performance assessment analyses

  10. Development of Pflotran Code for Waste Isolation Pilot Plant Performance Assessment

    Science.gov (United States)

    Zeitler, T.; Day, B. A.; Frederick, J.; Hammond, G. E.; Kim, S.; Sarathi, R.; Stein, E.

    2017-12-01

    The Waste Isolation Pilot Plant (WIPP) has been developed by the U.S. Department of Energy (DOE) for the geologic (deep underground) disposal of transuranic (TRU) waste. Containment of TRU waste at the WIPP is regulated by the U.S. Environmental Protection Agency (EPA). The DOE demonstrates compliance with the containment requirements by means of performance assessment (PA) calculations. WIPP PA calculations estimate the probability and consequence of potential radionuclide releases from the repository to the accessible environment for a regulatory period of 10,000 years after facility closure. The long-term performance of the repository is assessed using a suite of sophisticated computational codes. There is a current effort to enhance WIPP PA capabilities through the further development of the PFLOTRAN software, a state-of-the-art massively parallel subsurface flow and reactive transport code. Benchmark testing of the individual WIPP-specific process models implemented in PFLOTRAN (e.g., gas generation, chemistry, creep closure, actinide transport, and waste form) has been performed, including results comparisons for PFLOTRAN and existing WIPP PA codes. Additionally, enhancements to the subsurface hydrologic flow mode have been made. Repository-scale testing has also been performed for the modified PFLTORAN code and detailed results will be presented. Ultimately, improvements to the current computational environment will result in greater detail and flexibility in the repository model due to a move from a two-dimensional calculation grid to a three-dimensional representation. The result of the effort will be a state-of-the-art subsurface flow and transport capability that will serve WIPP PA into the future for use in compliance recertification applications (CRAs) submitted to the EPA. Sandia National Laboratories is a multi-mission laboratory managed and operated by National Technology and Engineering Solutions of Sandia, LLC., a wholly owned subsidiary of

  11. Evaluation of the Efficiency of Different Disruption Methods on Yeast Cell Wall Preparation for β-Glucan Isolation

    Directory of Open Access Journals (Sweden)

    Anna Bzducha-Wróbel

    2014-12-01

    Full Text Available Selected methods for yeast cell disruption were evaluated to establish their suitability for cell wall preparation in the process of β-glucan isolation. The effect of different disruption methods on contents of total saccharides, β-glucans and proteins in the produced cell walls preparations was analyzed. The degree of cell wall purification from intracellular components was established on the basis of the ratio of solubilised material. The investigated methods included: cell exposure to hot water (autoclaving, thermally-induced autolysis, homogenization in a bead mill, sonication and their combinations. Experimental systems were prepared in water (pH 5.0 and pH 7.0 and Tris-HCl buffer (pH 8.0. The Saccharomyces cerevisiae yeast cell wall preparations with the highest degree of cytosol component release and purification of β-glucans were produced by 30 min of cell homogenization with zirconium-glass beads (0.5 mm in diameter. This was confirmed by the highest ratio of solubilised material (approx. 64%–67%. The thus-produced preparations contained ca. 60% of total saccharides, 13%–14% of β(1,3/(1,6-glucans, and approx. 35% of crude proteins. Similar results were obtained after autolysis coupled with bead milling as well as with sonication, but the time required for these processes was more than 24 h. Homogenization in a bead mill could be valuable for general isolation procedures because allows one to eliminate the different autolytic activity of various yeast strains.

  12. Enhanced computational methods for quantifying the effect of geographic and environmental isolation on genetic differentiation

    DEFF Research Database (Denmark)

    Botta, Filippo; Eriksen, Casper; Fontaine, Michaël C.

    2015-01-01

    1. In a recent paper, Bradburd et al. (Evolution, 67, 2013, 3258) proposed a model to quantify the relative effect of geographic and environmental distance on genetic differentiation. Here, we enhance this method in several ways. 2. We modify the covariance model so as to fit better with mainstre...... available as an R package called sunder. It takes as input georeferenced allele counts at the individual or population level for co-dominant markers. Program homepage: http://www2.imm.dtu.dk/~gigu/Sunder/....

  13. A Polymerase Chain Reaction-Based Method for Isolating Clones from a Complimentary DNA Library in Sheep

    Science.gov (United States)

    Friis, Thor Einar; Stephenson, Sally; Xiao, Yin; Whitehead, Jon

    2014-01-01

    The sheep (Ovis aries) is favored by many musculoskeletal tissue engineering groups as a large animal model because of its docile temperament and ease of husbandry. The size and weight of sheep are comparable to humans, which allows for the use of implants and fixation devices used in human clinical practice. The construction of a complimentary DNA (cDNA) library can capture the expression of genes in both a tissue- and time-specific manner. cDNA libraries have been a consistent source of gene discovery ever since the technology became commonplace more than three decades ago. Here, we describe the construction of a cDNA library using cells derived from sheep bones based on the pBluescript cDNA kit. Thirty clones were picked at random and sequenced. This led to the identification of a novel gene, C12orf29, which our initial experiments indicate is involved in skeletal biology. We also describe a polymerase chain reaction-based cDNA clone isolation method that allows the isolation of genes of interest from a cDNA library pool. The techniques outlined here can be applied in-house by smaller tissue engineering groups to generate tools for biomolecular research for large preclinical animal studies and highlights the power of standard cDNA library protocols to uncover novel genes. PMID:24447069

  14. Isolated systems with wind power. Main report

    DEFF Research Database (Denmark)

    Lundsager, P.; Bindner, Henrik W.; Clausen, Niels-Erik

    2001-01-01

    The overall objective of this research project is to study the development of methods and guidelines rather than "universal solutions" for the use of wind energy in isolated communities. The main specific objective of the project is to develop and present amore unified and generally applicable...... approach for assessing the technical and economical feasibility of isolated power supply systems with wind energy. As a part of the project the following tasks were carried out: Review of literature, fieldmeasurements in Egypt, development of an inventory of small isolated systems, overview of end...... for Isolated Systems with Wind Power, applicable for international organisations such as donoragencies and development banks....

  15. Comparison of Some Extraction Methods for Isolation of Catechins and Caffeine from Turkish Green Tea

    Directory of Open Access Journals (Sweden)

    Ezgi DEMİR

    2015-07-01

    Full Text Available Effective extraction of anticancer and antioxidant principles from Turkish green tea were main purpose of this work. The pre-optimized experimental condition for liquid extraction was employed for comparative appraisal.  Not only extraction methods also nature of the green tea samples (fresh, dried or frozen and quantitative yields related to collection periods were investigated.  After extraction of the green tea with various techniques the extract was partitioned with chloroform to remove caffeine, after that the extract was partitioned with ethyl acetate to obtain catechin mixture. Quantification of individual catechins was carried out by HPLC and analysis results proved that epigallocatechin gallate (EGCG was main catechin specie present in all extracts. The results indicate that hot water extraction (at 80 0C provides higher catechin yield when compared to other methods. The highest extract yields were obtained with dried leaves collected in second collection period. The crude catechin mixture contains high amount of EGCG and might be used as raw material for production of plant remedies at industrial scale.

  16. Isolation of a somatomedin binding protein from human preterm amniotic fluid: development of a radioimmunoassay

    International Nuclear Information System (INIS)

    Drop, S.L.S.

    1983-01-01

    This thesis investigates the nature and biological behaviour of a somatomedin binding protein, identified in preterm amniotic fluid (AF). For that purpose a double antibody radioimmunoassay was developed. Purified AF binding protein (AFBP) was iodinated by the chloramine-T method, and dilutions of partially purified AFBP were designated as the standard, with the results expressed in μg equivalent protein/ml. The sensitivity of the assay was improved by adoption of the nonequilibrium procedure. AFBP values were twice as high in preterm AF as in term AF. (Auth.)

  17. Development of Convenient Screening Method for Resistant Radish to Plasmodiophora brassicae

    Directory of Open Access Journals (Sweden)

    Su-Jung Jo

    2011-08-01

    Full Text Available To establish simple and reliable screening method for resistant radish to Plasmodiophora brassicae Woron. using soil-drenching inoculation, the development of clubroot on radish seedlings inoculated with P. brassicae GN-1 isolate according to several conditions such as inoculum concentration, plant growth stage and incubation period after inoculation was studied. To select resistant radish against clubroot, 10-day-old seedlings were inoculated with P. brassicae by drenching the roots with the spore suspension of the pathogen to give 1×10(9 spores/pot. The inoculated seedlings were incubated in a growth chamber at 20℃ for 3 days then cultivated in a greenhouse (20±5℃ for 6 weeks. Under the optimum conditions, 46 commercial cultivars of radish were tested for resistance to YC-1 (infecting 15 clubroot-resistant cultivars of Chinese cabbage and GN-1 (wild type isolates of P. brassicae. Among them, thirty-five cultivars showed resistance to both isolates and one cultivar represented susceptible response to the pathogens. On the other hand, the other cultivars showed different responses against the tested P. brassicae pathogens. The results suggest that this method is an efficient system for screening radish with resistance to clubroot.

  18. Development of a community's self-efficacy scale for preventing social isolation among community-dwelling older people (Mimamori Scale).

    Science.gov (United States)

    Tadaka, Etsuko; Kono, Ayumi; Ito, Eriko; Kanaya, Yukiko; Dai, Yuka; Imamatsu, Yuki; Itoi, Waka

    2016-11-28

    Among older people in developed countries, social isolation leading to solitary death has become a public health issue of vital importance. Such isolation could be prevented by monitoring at-risk individuals at the neighborhood level and by implementing supportive networks at the community level. However, a means of measuring community confidence in these measures has not been established. This study is aimed at developing the Community's Self-Efficacy Scale (CSES; Mimamori scale in Japanese) for community members preventing social isolation among older people. The CSES is a self-administered questionnaire developed on the basis of Bandura's self-efficacy theory. The survey was given to a general population (GEN) sample (n = 6,000) and community volunteer (CVOL) sample (n = 1,297). Construct validity was determined using confirmatory factor analysis. Internal consistency was calculated using Cronbach's alpha. The Generative Concern Scale (GCS-R) and Brief Sense of Community Scale (BSCS) were also administered to assess criterion-related validity of the CSES. In total, 3,484 and 859 valid responses were received in the GEN and CVOL groups, respectively. The confirmatory factor analysis identified eight items from two domains-community network and neighborhood watch-with goodness of fit index = 0.984, adjusted goodness of fit index = 0.970, comparative fit index = 0.988, and root mean square error of approximation = 0.047. Cronbach's alpha for the entire CSES was 0.87 and for the subscales was 0.80 and higher. The score of the entire CSES was positively correlated with the GCS-R in both the GEN (r = 0.80, p social isolation among older people. The scale is potentially useful for promoting health policies, practices, and interventions within communities. This may help prevent social isolation among older people and contribute to overall well-being in aging societies in Japan and abroad.

  19. Effects of extracellular matrices and growth factors on the development of isolated porcine blastomeres.

    Science.gov (United States)

    Saito, S; Niemann, H

    1991-05-01

    The effects of extracellular matrices and growth factors on the development of isolated blastomeres derived from intact 4-, 8-, and 16-cell porcine embryos (termed, respectively, 1/4, 1/8, and 1/16 blastomeres) were investigated in vitro and in vivo. Blastomeres were incubated in extracellular matrix components fibronectin (FIN) or swine skin gelatin (SSG)-precoated culture dishes containing either modified Krebs' Ringer Bicarbonate solution (mKRB) supplemented with 10% heat-inactivated lamb serum, or Hanks' solution supplemented with 10% heat-inactivated newborn calf serum (NBCS) or Waymouth medium supplemented with 10% NBCS or in noncoated dishes in mKRB supplemented with either insulin (10, 100, or 1,000 micrograms/ml), transferrin (10, 100, or 1,000 micrograms/ml), or cAMP (0.2 or 2.0 micrograms/ml). Cultures observed at 24-h intervals and morphological development was recorded. Blastomeres were classified into three categories according to their morphology: (1) regular blastocysts, (2) trophectodermal vesicles, or (3) no development. After 96 h, culture was determined; the overall diameter of the blastocysts was determined and the nuclei were counted. Blastomeres/blastocysts did not adhere to the bottom of the culture dishes coated with extracellular matrices. Blastocyst formation rate was highest when FIN/mKRB was used and reached 44.3%, 41.8%, and 36.5% for 1/4, 1/8, and 1/16 blastomeres, respectively. The respective blastocysts contained an average of 31.2 +/- 5.8, 58.2 +/- 8.4, and 18.5 +/- 3.5 nuclei and had an overall diameter of 250.0 +/- 10.1, 235.0 +/- 12.8, and 172.5 +/- 13.7 microns, 1/8 blastomeres displayed a better (p less than 0.05) growth rate than 1/4 and 1/16 blastomeres, and 1/8 blastomeres in FIN/mKRB grew better (p less than 0.01) when cultured in an open system than in a microdrop under oil (35.5% vs. 5.0% blastocysts). Neither cAMP nor transferrin had a significant stimulating effect on blastocyst development of 1/8 blastomeres when m

  20. A comparative study of thermal calcination and an alkaline hydrolysis method in the isolation of hydroxyapatite from Thunnus obesus bone

    International Nuclear Information System (INIS)

    Venkatesan, Jayachandran; Ryu, BoMi; Thomas, Noel Vinay; Kim, Se Kwon; Qian Zhongji

    2011-01-01

    In the present study, hydroxyapatite (HAp) was isolated from Thunnus obesus bone using alkaline hydrolysis and thermal calcination methods. The obtained ceramic has been characterized by thermal gravimetric analysis (TGA), Fourier transform infrared spectroscopy (FT-IR), powder x-ray diffraction analysis (XRD), field-emission scanning electron microscopy, energy-dispersive x-ray analysis, transmission electron microscopy (TEM), selected area diffraction analysis, cytotoxic analysis and cell proliferation analysis. The results indicate that there are significant differences between the ceramics and T. obesus bone. FT-IR and TGA results affirmed that the collagen and organic moieties have been eliminated by both the proposed methods. XRD results were in agreement with JCPDS data. TEM and selective area diffraction images have signified that the thermal calcination method produces good crystallinity with dimensions 0.3-1.0 μm, whereas the alkaline hydrolysis method produces nanostructured HAp crystals with 17-71 nm length and 5-10 nm width. Biocompatibility of HAp crystals was evaluated by cytotoxicity and cell proliferation with human osteoblast-like cell MG-63.