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Sample records for flow cytometric nuclear

  1. Comparison of Five Nuclear Isolation Buffers for Flow Cytometric ...

    African Journals Online (AJOL)

    Jocky

    2012-02-23

    Feb 23, 2012 ... factor as a measure of sample quality (%DF) and nuclear yield factor (YF) in order to compare the quantity of nuclei in suspension. The following equations were used: Estimation of DNA contents for cultivars classification. Young leaves of Deli Dura, Pisifera and hybrid Tenera trees were prepared following ...

  2. Simultaneous flow cytometric quantification of plant nuclear DNA contents over the full range of described angiosperm 2C values.

    Science.gov (United States)

    Galbraith, David W

    2009-08-01

    Flow cytometry provides a rapid, accurate, and simple means to determine nuclear DNA contents (C-value) within plant homogenates. This parameter is extremely useful in a number of applications in basic and applied plant biology; for example, it provides an important starting point for projects involving whole genome sequencing, it facilitates characterization of plant species within natural and agricultural settings, it allows facile identification of engineered plants that are euploid or that represent desired ploidy classes, it points toward studies concerning the role of C-value in plant growth and development and in response to the environment and in terms of evolutionary fitness, and, in uncovering new and unexpected phenomena (for example endoreduplication), it uncovers new avenues of scientific enquiry. Despite the ease of the method, C-values have been determined for only around 2% of the described angiosperm (flowering plant) species. Within this small subset, one of the most remarkable observations is the range of 2C values, which spans at least two orders of magnitude. In determining C-values for new species, technical issues are encountered which relate both to requirement for a method that can provide accurate measurements across this extended dynamic range, and that can accommodate the large amounts of debris which accompanies flow measurements of plant homogenates. In this study, the use of the Accuri C6 flow cytometer for the analysis of plant C-values is described. This work indicates that the unusually large dynamic range of the C6, a design feature, coupled to the linearity of fluorescence emission conferred by staining of nuclei using propidium iodide, allows simultaneous analysis of species whose C-values span that of almost the entire described angiosperms. Copyright 2009 International Society for Advancement of Cytometry.

  3. Nuclear chromatin variations in human spermatozoa undergoing swim-up and cryopreservation evaluated by the flow cytometric sperm chromatin structure assay.

    Science.gov (United States)

    Spanò, M; Cordelli, E; Leter, G; Lombardo, F; Lenzi, A; Gandini, L

    1999-01-01

    The sperm chromatin structure assay (SCSA) is a flow cytometric (FCM) technique which exploits the metachromatic properties of Acridine Orange to monitor the susceptibility of sperm chromatin DNA to in-situ acid denaturation. SCSA was used to study the chromatin structure variations of human spermatozoa in semen, both before and after swim-up and after cryopreservation. Semen samples were provided by 19 healthy normozoospermic subjects attending pre-marriage checks. Each sample was divided into three aliquots: the first aliquot was evaluated without further treatment, the second underwent swim-up, and the third was stored according to standard cryopreservation techniques in liquid nitrogen at -196 degrees C. Samples were also analysed by light and fluorescence microscopy (after Acridine Orange staining to evaluate the number of green fluorescent sperm heads), and by computer-assisted semen analysis. The results showed that post-rise spermatozoa represent a subpopulation characterized by a general improvement of the morphological (reduction of the percentage of abnormal forms and heads, increase of the green head sperm percentage) and kinetic parameters. This subpopulation also exhibited improved chromatin structure properties, confirming that these cells have the best structural and functional characteristics, indicative of optimal fertilizing ability. On the other hand, overall sperm quality deteriorates after cryopreservation. When thawed spermatozoa underwent an additional swim-up round, a general improvement of nuclear maturity was seen in the post-rise spermatozoa.

  4. Challenges of flow-cytometric estimation of nuclear genome size in orchids, a plant group with both whole-genome and progressively partial endoreplication.

    Science.gov (United States)

    Trávníček, Pavel; Ponert, Jan; Urfus, Tomáš; Jersáková, Jana; Vrána, Jan; Hřibová, Eva; Doležel, Jaroslav; Suda, Jan

    2015-10-01

    Nuclear genome size is an inherited quantitative trait of eukaryotic organisms with both practical and biological consequences. A detailed analysis of major families is a promising approach to fully understand the biological meaning of the extensive variation in genome size in plants. Although Orchidaceae accounts for ∼10% of the angiosperm diversity, the knowledge of patterns and dynamics of their genome size is limited, in part due to difficulties in flow cytometric analyses. Cells in various somatic tissues of orchids undergo extensive endoreplication, either whole-genome or partial, and the G1-phase nuclei with 2C DNA amounts may be lacking, resulting in overestimated genome size values. Interpretation of DNA content histograms is particularly challenging in species with progressively partial endoreplication, in which the ratios between the positions of two neighboring DNA peaks are lower than two. In order to assess distributions of nuclear DNA amounts and identify tissue suitable for reliable estimation of nuclear DNA content, we analyzed six different tissue types in 48 orchid species belonging to all recognized subfamilies. Although traditionally used leaves may provide incorrect C-values, particularly in species with progressively partial endoreplication, young ovaries and pollinaria consistently yield 2C and 1C peaks of their G1-phase nuclei, respectively, and are, therefore, the most suitable parts for genome size studies in orchids. We also provide new DNA C-values for 22 orchid genera and 42 species. Adhering to the proposed methodology would allow for reliable genome size estimates in this largest plant family. Although our research was limited to orchids, the need to find a suitable tissue with dominant 2C peak of G1-phase nuclei applies to all endopolyploid species. © 2015 International Society for Advancement of Cytometry.

  5. Flow cytometric analysis of variation in the level of nuclear DNA endoreduplication in the cotyledons amongst Vigna radiata cultivars

    Czech Academy of Sciences Publication Activity Database

    Pal, A.; Vrána, Jan; Doležel, Jaroslav

    2004-01-01

    Roč. 57, č. 3 (2004), s. 262-266 ISSN 0008-7114 R&D Projects: GA AV ČR IBS5038104 Grant - others:Indian National Science Academy(IN) INSA Institutional research plan: CEZ:AV0Z5038910 Keywords : Cotyledon * endoreduplication * flow cytometry Subject RIV: GE - Plant Breeding Impact factor: 0.366, year: 2004

  6. A flow cytometric assay for simultaneously measuring the ...

    African Journals Online (AJOL)

    Jane

    2011-10-24

    Oct 24, 2011 ... This research objective was to exploit a novel method for measuring the proliferation, cytotoxicity of cytokine-induced killer (CIK) cells using carboxyfluorescein succinimidyl ester/proliferation index. (CFSE/PI) and flow cytometric assay. As cells divide, CFSE is apportioned equally between the two daughter ...

  7. Flow cytometric immunoassay for sulfonamides in raw milk

    NARCIS (Netherlands)

    Keizer, de W.; Bienenmann-Ploum, M.; Bergwerff, A.A.; Haasnoot, W.

    2008-01-01

    Sulfonamide antibiotics are applied in veterinary medicine for the treatment of microbial infections. For the detection of residues of sulfonamides in milk, a multi-sulfonamide flow cytometric immunoassay (FCI) was developed using the Luminex MultiAnalyte Profiling (xMAP) technology. In this

  8. A flow cytometric assay for simultaneously measuring the ...

    African Journals Online (AJOL)

    This research objective was to exploit a novel method for measuring the proliferation, cytotoxicity of cytokine-induced killer (CIK) cells using carboxyfluorescein succinimidyl ester/proliferation index (CFSE/PI) and flow cytometric assay. As cells divide, CFSE is apportioned equally between the two daughter cells, leading to a ...

  9. Technical discussions II - Flow cytometric analysis

    NARCIS (Netherlands)

    Cunningham, A; Cid, A; Buma, AGJ

    In this paper the potencial of flow cytometry as applied to the aquatic life sciences is discussed. The use of flow cytometry for studying the ecotoxicology of phytoplankton was introduced. On the other hand, the new flow cytometer EUROPA was presented. This is a multilaser machine which has been

  10. Flow cytometric fingerprinting for microbial strain discrimination and physiological characterization.

    Science.gov (United States)

    Buysschaert, Benjamin; Kerckhof, Frederiek-Maarten; Vandamme, Peter; De Baets, Bernard; Boon, Nico

    2018-02-01

    The analysis of microbial populations is fundamental, not only for developing a deeper understanding of microbial communities but also for their engineering in biotechnological applications. Many methods have been developed to study their characteristics and over the last few decades, molecular analysis tools, such as DNA sequencing, have been used with considerable success to identify the composition of microbial populations. Recently, flow cytometric fingerprinting is emerging as a promising and powerful method to analyze bacterial populations. So far, these methods have primarily been used to observe shifts in the composition of microbial communities of natural samples. In this article, we apply a flow cytometric fingerprinting method to discriminate among 29 Lactobacillus strains. Our results indicate that it is possible to discriminate among 27 Lactobacillus strains by staining with SYBR green I and that the discriminatory power can be increased by combined SYBR green I and propidium iodide staining. Furthermore, we illustrate the impact of physiological changes on the fingerprinting method by demonstrating how flow cytometric fingerprinting is able to discriminate the different growth phases of a microbial culture. The sensitivity of the method is assessed by its ability to detect changes in the relative abundance of a mix of polystyrene beads down to 1.2%. When a mix of bacteria was used, the sensitivity was as between 1.2% and 5%. The presented data demonstrate that flow cytometric fingerprinting is a sensitive and reproducible technique with the potential to be applied as a method for the dereplication of bacterial isolates. © 2017 International Society for Advancement of Cytometry. © 2017 International Society for Advancement of Cytometry.

  11. Uncovering Aberrant Mutant PKA Function with Flow Cytometric FRET

    Directory of Open Access Journals (Sweden)

    Shin-Rong Lee

    2016-03-01

    Full Text Available Biology has been revolutionized by tools that allow the detection and characterization of protein-protein interactions (PPIs. Förster resonance energy transfer (FRET-based methods have become particularly attractive as they allow quantitative studies of PPIs within the convenient and relevant context of living cells. We describe here an approach that allows the rapid construction of live-cell FRET-based binding curves using a commercially available flow cytometer. We illustrate a simple method for absolutely calibrating the cytometer, validating our binding assay against the gold standard isothermal calorimetry (ITC, and using flow cytometric FRET to uncover the structural and functional effects of the Cushing-syndrome-causing mutation (L206R on PKA’s catalytic subunit. We discover that this mutation not only differentially affects PKAcat’s binding to its multiple partners but also impacts its rate of catalysis. These findings improve our mechanistic understanding of this disease-causing mutation, while illustrating the simplicity, general applicability, and power of flow cytometric FRET.

  12. Automated High-Dimensional Flow Cytometric Data Analysis

    Science.gov (United States)

    Pyne, Saumyadipta; Hu, Xinli; Wang, Kui; Rossin, Elizabeth; Lin, Tsung-I.; Maier, Lisa; Baecher-Allan, Clare; McLachlan, Geoffrey; Tamayo, Pablo; Hafler, David; de Jager, Philip; Mesirov, Jill

    Flow cytometry is widely used for single cell interrogation of surface and intracellular protein expression by measuring fluorescence intensity of fluorophore-conjugated reagents. We focus on the recently developed procedure of Pyne et al. (2009, Proceedings of the National Academy of Sciences USA 106, 8519-8524) for automated high- dimensional flow cytometric analysis called FLAME (FLow analysis with Automated Multivariate Estimation). It introduced novel finite mixture models of heavy-tailed and asymmetric distributions to identify and model cell populations in a flow cytometric sample. This approach robustly addresses the complexities of flow data without the need for transformation or projection to lower dimensions. It also addresses the critical task of matching cell populations across samples that enables downstream analysis. It thus facilitates application of flow cytometry to new biological and clinical problems. To facilitate pipelining with standard bioinformatic applications such as high-dimensional visualization, subject classification or outcome prediction, FLAME has been incorporated with the GenePattern package of the Broad Institute. Thereby analysis of flow data can be approached similarly as other genomic platforms. We also consider some new work that proposes a rigorous and robust solution to the registration problem by a multi-level approach that allows us to model and register cell populations simultaneously across a cohort of high-dimensional flow samples. This new approach is called JCM (Joint Clustering and Matching). It enables direct and rigorous comparisons across different time points or phenotypes in a complex biological study as well as for classification of new patient samples in a more clinical setting.

  13. A flow cytometric technique for quantification and differentiation of bacteria in bulk tank milk

    DEFF Research Database (Denmark)

    Holm, C.; Mathiasen, T.; Jespersen, Lene

    2004-01-01

    AIMS: The present study describes a flow cytometric technique for quantification and differentiation of bacteria in bulk tank milk according to the main cause of elevated counts. METHODS AND RESULTS: A total of 75 Danish bulk tank milk samples exceeding the grading level of 3.0 x 10(4) CFU ml(-1...... parameters were as follows: staining with Oregon Green conjugated wheat germ agglutinin that binds to the cell wall of bacteria, staining with hexidium iodide that binds to all bacterial DNA, the flow cytometric forward scatter and the flow cytometric side scatter. Three regions in the flow cytometric plot...

  14. Flow Cytometric Bead Sandwich Assay Based on a Split Aptamer.

    Science.gov (United States)

    Shen, Luyao; Bing, Tao; Liu, Xiangjun; Wang, Junyan; Wang, Linlin; Zhang, Nan; Shangguan, Dihua

    2018-01-24

    A few aptamers still bind their targets after being split into two moieties. Split aptamers have shown great potential in the development of aptameric sensors. However, only a few split aptamers have been generated because of lack of knowledge on the binding structure of their parent aptamers. Here, we report the design of a new split aptamer and a flow cytometric bead sandwich assay using a split aptamer instead of double antibodies. Through DMS footprinting and mutation assay, we figured out the target-binding moiety and the structure-stabilizing moiety of the l-selectin aptamer, Sgc-3b. By separating the duplex strand in the structure-stabilizing moiety, we obtained a split aptamer that bound l-selectin. After optimization of one part of the split sequence to eliminate the nonspecific binding of the split sequence pair, we developed a split-aptamer-based cytometric bead assay (SACBA) for the detection of soluble l-selectin. SACBA showed good sensitivity and selectivity to l-selectin and was successfully applied for the detection of spiked l-selectin in the human serum. The strategies for generating split aptamers and designing the split-aptamer-based sandwich assay are simple and efficient and show good practicability in aptamer engineering.

  15. Multivariate analysis of flow cytometric data using decision trees.

    Science.gov (United States)

    Simon, Svenja; Guthke, Reinhard; Kamradt, Thomas; Frey, Oliver

    2012-01-01

    Characterization of the response of the host immune system is important in understanding the bidirectional interactions between the host and microbial pathogens. For research on the host site, flow cytometry has become one of the major tools in immunology. Advances in technology and reagents allow now the simultaneous assessment of multiple markers on a single cell level generating multidimensional data sets that require multivariate statistical analysis. We explored the explanatory power of the supervised machine learning method called "induction of decision trees" in flow cytometric data. In order to examine whether the production of a certain cytokine is depended on other cytokines, datasets from intracellular staining for six cytokines with complex patterns of co-expression were analyzed by induction of decision trees. After weighting the data according to their class probabilities, we created a total of 13,392 different decision trees for each given cytokine with different parameter settings. For a more realistic estimation of the decision trees' quality, we used stratified fivefold cross validation and chose the "best" tree according to a combination of different quality criteria. While some of the decision trees reflected previously known co-expression patterns, we found that the expression of some cytokines was not only dependent on the co-expression of others per se, but was also dependent on the intensity of expression. Thus, for the first time we successfully used induction of decision trees for the analysis of high dimensional flow cytometric data and demonstrated the feasibility of this method to reveal structural patterns in such data sets.

  16. Oxidative product formation in irradiated neutrophils. A flow cytometric analysis

    International Nuclear Information System (INIS)

    Wolber, R.A.; Duque, R.E.; Robinson, J.P.; Oberman, H.A.

    1987-01-01

    The effect of irradiation on neutrophil oxidative function was evaluated using a flow cytometric assay of intracellular hydrogen peroxide (H 2 O 2 ) production. This assay quantitates the H 2 O 2 -dependent conversion of the nonfluorescent compound, 2'-7'-dichlorofluorescein (DCFH), into fluorescent 2'-7'-dichlorofluorescein (DCF) on a single-cell basis. Intracellular H 2 O 2 production in response to stimulation with phorbol myristate acetate was not affected by neutrophil irradiation at doses up to 2500 rad. In addition, irradiation of intracellular DCFH and aqueous 2'-7'-dichlorofluorescein diacetate (DCFH-DA) resulted in DCF production, which suggested that oxidative molecules produced by aqueous radiolysis were detected by this assay. This study indicates that radiation doses of 1500 to 2500 rad, which are sufficient to prevent induction of graft-versus-host disease by transfused blood components, are not deleterious to neutrophil oxidative metabolism

  17. Discrimination of bromodeoxyuridine labelled and unlabelled mitotic cells in flow cytometric bromodeoxyuridine/DNA analysis

    DEFF Research Database (Denmark)

    Jensen, P O; Larsen, J K; Christensen, I J

    1994-01-01

    Bromodeoxyuridine (BrdUrd) labelled and unlabelled mitotic cells, respectively, can be discriminated from interphase cells using a new method, based on immunocytochemical staining of BrdUrd and flow cytometric four-parameter analysis of DNA content, BrdUrd incorporation, and forward and orthogonal...... light scatter. The method was optimized using the human leukemia cell lines HL-60 and K-562. Samples of 10(5) ethanol-fixed cells were treated with pepsin/HCl and stained as a nuclear suspension with anti-BrdUrd antibody, FITC-conjugated secondary antibody, and propidium iodide. Labelled mitoses could...

  18. The cytometric future: it ain't necessarily flow!

    Science.gov (United States)

    Shapiro, Howard M

    2011-01-01

    Initial approaches to cytometry for classifying and characterizing cells were based on microscopy; it was necessary to collect relatively high-resolution images of cells because only a few specific reagents usable for cell identification were available. Although flow cytometry, now the dominant cytometric technology, typically utilizes lenses similar to microscope lenses for light collection, improved, more quantitative reagents allow the necessary information to be acquired in the form of whole-cell measurements of the intensities of light transmission, scattering, and/or fluorescence.Much of the cost and complexity of both automated microscopes and flow cytometers arises from the necessity for them to measure one cell at a time. Recent developments in digital camera technology now offer an alternative in which one or more low-magnification, low-resolution images are made of a wide field containing many cells, using inexpensive light-emitting diodes (LEDs) for illumination. Minimalist widefield imaging cytometers can provide a smaller, less complex, and substantially less expensive alternative to flow cytometry, critical in systems intended for in resource-poor areas. Minimalism is, likewise, a good philosophy in developing instrumentation and methodology for both clinical and large-scale research use; it simplifies quality assurance and compliance with regulatory requirements, as well as reduces capital outlays, material costs, and personnel training requirements. Also, importantly, it yields "greener" technology.

  19. Detection of circulating immune complexes by Raji cell assay: comparison of flow cytometric and radiometric methods

    International Nuclear Information System (INIS)

    Kingsmore, S.F.; Crockard, A.D.; Fay, A.C.; McNeill, T.A.; Roberts, S.D.; Thompson, J.M.

    1988-01-01

    Several flow cytometric methods for the measurement of circulating immune complexes (CIC) have recently become available. We report a Raji cell flow cytometric assay (FCMA) that uses aggregated human globulin (AHG) as primary calibrator. Technical advantages of the Raji cell flow cytometric assay are discussed, and its clinical usefulness is evaluated in a method comparison study with the widely used Raji cell immunoradiometric assay. FCMA is more precise and has greater analytic sensitivity for AHG. Diagnostic sensitivity by the flow cytometric method is superior in systemic lupus erythematosus (SLE), rheumatoid arthritis, and vasculitis patients: however, diagnostic specificity is similar for both assays, but the reference interval of FCMA is narrower. Significant correlations were found between CIC levels obtained with both methods in SLE, rheumatoid arthritis, and vasculitis patients and in longitudinal studies of two patients with cerebral SLE. The Raji cell FCMA is recommended for measurement of CIC levels to clinical laboratories with access to a flow cytometer

  20. Detection of circulating immune complexes by Raji cell assay: comparison of flow cytometric and radiometric methods

    Energy Technology Data Exchange (ETDEWEB)

    Kingsmore, S.F.; Crockard, A.D.; Fay, A.C.; McNeill, T.A.; Roberts, S.D.; Thompson, J.M.

    1988-01-01

    Several flow cytometric methods for the measurement of circulating immune complexes (CIC) have recently become available. We report a Raji cell flow cytometric assay (FCMA) that uses aggregated human globulin (AHG) as primary calibrator. Technical advantages of the Raji cell flow cytometric assay are discussed, and its clinical usefulness is evaluated in a method comparison study with the widely used Raji cell immunoradiometric assay. FCMA is more precise and has greater analytic sensitivity for AHG. Diagnostic sensitivity by the flow cytometric method is superior in systemic lupus erythematosus (SLE), rheumatoid arthritis, and vasculitis patients: however, diagnostic specificity is similar for both assays, but the reference interval of FCMA is narrower. Significant correlations were found between CIC levels obtained with both methods in SLE, rheumatoid arthritis, and vasculitis patients and in longitudinal studies of two patients with cerebral SLE. The Raji cell FCMA is recommended for measurement of CIC levels to clinical laboratories with access to a flow cytometer.

  1. Challenges of flow-cytometric estimation of nuclear genome size in orchids, a plant group with both whole-genome and progressively partial endoreplication

    Czech Academy of Sciences Publication Activity Database

    Trávníček, Pavel; Ponert, J.; Urfus, Tomáš; Jersáková, Jana; Vrána, Jan; Hřibová, Eva; Doležel, Jaroslav; Suda, Jan

    2015-01-01

    Roč. 87, č. 10 (2015), s. 958-966 ISSN 1552-4922 R&D Projects: GA ČR GAP506/12/1320 Institutional support: RVO:67985939 ; RVO:67179843 ; RVO:61389030 Keywords : flow cytometry * genome size * hyporeduplication Subject RIV: EF - Botanics; EF - Botanics (UEB-Q); EH - Ecology, Behaviour (UEK-B) Impact factor: 3.181, year: 2015

  2. Flow cytometric analysis using SYBR Green I for genome size estimation in coffee.

    Science.gov (United States)

    Ronildo Clarindo, Wellington; Roberto Carvalho, Carlos

    2011-02-01

    Plant genome size has been measured by flow cytometry using propidium iodide as a dye for nuclear DNA staining. However, some authors have reported the occurrence of genome size estimation errors, especially in plants rich in secondary metabolites, such as the coffee tree. In this context, we tested an alternative cytometric protocol using the SYBR Green I as a fluorochrome for stoichiometrically staining nuclear double-stranded DNA in Coffea canephora (2x) and Coffea arabica (4x). The results showed that the respective mean genome size measured from nuclei stained with SYBR Green I and propidium iodide was statistically identical. However, the G(0)/G(1) peaks of nuclei stained with SYBR Green I exhibited lower coefficient variations (1.57-2.85%) compared to those stained with propidium iodide (2.75-4.80%). Coefficient variation statistical data suggest that SYBR Green I is adequate for stoichiometric nuclei staining using this methodology. Our results provide evidence that SYBR Green I can be used in flow cytometry measurements of plants, with the advantages of minimizing errors in nuclear DNA content quantification, staining relatively quicker, with high affinity, and being less mutagenic than propidium iodide. Copyright © 2009 Elsevier GmbH. All rights reserved.

  3. Flow cytometric DNA analysis of ducks accumulating 137Cs on a reactor reservoir

    International Nuclear Information System (INIS)

    George, L.S.; Dallas, C.E.; Brisbin, I.L. Jr.; Evans, D.L.

    1991-01-01

    The objective of this study was to detect red blood cell (rbc) DNA abnormalities in male, game-farm mallard ducks as they ranged freely and accumulated 137Cs (radiocesium) from an abandoned nuclear reactor cooling reservoir. Prior to release, the ducks were tamed to enable recapture at will. Flow cytometric measurements conducted at intervals during the first year of exposure yielded cell cycle percentages of DNA (G0/G1, S, G2 + M phases) of rbc, as well as coefficients of variation (CV) in the G0/G1 phase. DNA histograms of exposed ducks were compared with two sets of controls which were maintained 30 and 150 miles from the study site. 137Cs live wholebody burdens were also measured in these animals in a parallel kinetics study, and an approximate steady-state equilibrium was attained after about 8 months. DNA histograms from 2 of the 14 contaminated ducks revealed DNA aneuploid-like patterns after 9 months exposure. These two ducks were removed from the experiment at this time, and when sampled again 1 month later, one continued to exhibit DNA aneuploidy. None of the control DNA histograms demonstrated DNA aneuploid-like patterns. There were no significant differences in cell cycle percentages at any time point between control and exposed animals. A significant increase in CV was observed at 9 months exposure, but after removal of the two ducks with DNA aneuploidy, no significant difference was detected in the group monitored after 12 months exposure. An increased variation in the DNA and DNA aneuploidy could, therefore, be detected in duck rbc using flow cytometric analysis, with the onset of these effects being related to the attainment of maximal levels of 137Cs body burdens in the exposed animals

  4. Activity of ethanol-stressed Oenococcus oeni cells: a flow cytometric approach

    NARCIS (Netherlands)

    Silveira, da M.G.; Abee, T.

    2009-01-01

    Aims: To study the effect of ethanol on Oenococcus oeni activity at the single cell level. Methods and Results: The active extrusion of the fluorescent probe carboxy fluorescein (cF) was used to assess the metabolic activity of ethanol-stressed O. oeni cells. Subsequent flow cytometric analysis

  5. Double fluorescent flow cytometric assessment of bacterial internalization and binding by epithelial cells

    NARCIS (Netherlands)

    de Boer, E. C.; Bevers, R. F.; Kurth, K. H.; Schamhart, D. H.

    1996-01-01

    This study describes a new flow cytometric method for assessment of phagocytosis of specific bacteria (bacillus Calmette-Guérin (BCG) and Escherichia coli) by bladder epithelial cells. The internalization assay consisted of labeling bacteria chemically with fluorescein isothiocyanate (FITC).

  6. Cluster Analysis of Flow Cytometric List Mode Data on a Personal Computer

    NARCIS (Netherlands)

    Bakker Schut, Tom C.; Bakker schut, T.C.; de Grooth, B.G.; Greve, Jan

    1993-01-01

    A cluster analysis algorithm, dedicated to analysis of flow cytometric data is described. The algorithm is written in Pascal and implemented on an MS-DOS personal computer. It uses k-means, initialized with a large number of seed points, followed by a modified nearest neighbor technique to reduce

  7. Intrinsic properties of so-called dormant probiotic bacteria, determined by flow cytometric viability assays.

    Science.gov (United States)

    Lahtinen, Sampo J; Ouwehand, Arthur C; Reinikainen, Johanna P; Korpela, Jaakko M; Sandholm, Jouko; Salminen, Seppo J

    2006-07-01

    Plate counting and four culture-independent flow cytometric assays were used to determine the viability and intrinsic properties of three probiotic strains during storage. The strains showed reduction in plate counts but were able to maintain esterase activity, intact cytoplasmic membrane, and pH gradient. The apparently uncultivable probiotic cells were active and stress resistant.

  8. Intrinsic Properties of So-Called Dormant Probiotic Bacteria, Determined by Flow Cytometric Viability Assays

    OpenAIRE

    Lahtinen, Sampo J.; Ouwehand, Arthur C.; Reinikainen, Johanna P.; Korpela, Jaakko M.; Sandholm, Jouko; Salminen, Seppo J.

    2006-01-01

    Plate counting and four culture-independent flow cytometric assays were used to determine the viability and intrinsic properties of three probiotic strains during storage. The strains showed reduction in plate counts but were able to maintain esterase activity, intact cytoplasmic membrane, and pH gradient. The apparently uncultivable probiotic cells were active and stress resistant.

  9. Flow cytometric method for measuring chromatin fragmentation in fixed sperm from yellow perch (Perca flavescens)

    Science.gov (United States)

    Jenkins, Jill A.; Draugelis-Dale, Rassa O.; Pinkney, Alfred E.; Iwanowicz, Luke R.; Blazer, Vicki

    2015-01-01

    Declining harvests of yellow perch, Perca flavescens, in urbanized watersheds of Chesapeake Bay have prompted investigations of their reproductive fitness. The purpose of this study was to establish a flow cytometric technique for DNA analysis of fixed samples sent from the field to provide reliable gamete quality measurements. Similar to the sperm chromatin structure assay, measures were made on the susceptibility of nuclear DNA to acid-induced denaturation, but used fixed rather than live or thawed cells. Nuclei were best exposed to the acid treatment for 1 minute at 37 °C followed by the addition of cold (4 °C) propidium iodide staining solution before flow cytometry. The rationale for protocol development is presented graphically through cytograms. Field results collected in 2008 and 2009 revealed DNA fragmentation up to 14.5%. In 2008, DNA fragmentation from the more urbanized watersheds was significantly greater than from reference sites (P = 0.026) and in 2009, higher percentages of haploid testicular cells were noted from the less urbanized watersheds (P = 0.032) indicating better reproductive condition at sites with less urbanization. For both years, total and progressive live sperm motilities by computer-assisted sperm motion analysis ranged from 19.1% to 76.5%, being significantly higher at the less urbanized sites (P chromatin compaction levels from samples translocated over distance and time. The protocol provides an approach that can be modified for other species across taxa.

  10. DNA flow cytometric analysis in variable types of hydropic placentas

    Directory of Open Access Journals (Sweden)

    Fatemeh Atabaki pasdar

    2015-05-01

    Full Text Available Background: Differential diagnosis between complete hydatidiform mole, partial hydatidiform mole and hydropic abortion, known as hydropic placentas is still a challenge for pathologists but it is very important for patient management. Objective: We analyzed the nuclear DNA content of various types of hydropic placentas by flowcytometry. Materials and Methods: DNA ploidy analysis was performed in 20 non-molar (hydropic and non-hydropic spontaneous abortions and 20 molar (complete and partial moles, formalin-fixed, paraffin-embedded tissue samples by flow cytometry. The criteria for selection were based on the histopathologic diagnosis. Results: Of 10 cases histologically diagnosed as complete hydatiform mole, 9 cases yielded diploid histograms, and 1 case was tetraploid. Of 10 partial hydatidiform moles, 8 were triploid and 2 were diploid. All of 20 cases diagnosed as spontaneous abortions (hydropic and non-hydropic yielded diploid histograms. Conclusion: These findings signify the importance of the combined use of conventional histology and ploidy analysis in the differential diagnosis of complete hydatidiform mole, partial hydatidiform mole and hydropic abortion.

  11. Image cytometric nuclear texture features in inoperable head and neck cancer: a pilot study

    International Nuclear Information System (INIS)

    Strojan-Flezar, Margareta; Lavrencak, Jaka; Zganec, Mario; Strojan, Primoz

    2011-01-01

    Image cytometry can measure numerous nuclear features which could be considered a surrogate end-point marker of molecular genetic changes in a nucleus. The aim of the study was to analyze image cytometric nuclear features in paired samples of primary tumor and neck metastasis in patients with inoperable carcinoma of the head and neck. Image cytometric analysis of cell suspensions prepared from primary tumor tissue and fine needle aspiration biopsy cell samples of neck metastases from 21 patients treated with concomitant radiochemotherapy was performed. Nuclear features were correlated with clinical characteristics and response to therapy. Manifestation of distant metastases and new primaries was associated (p<0.05) with several chromatin characteristics from primary tumor cells, whereas the origin of index cancer and disease response in the neck was related to those in the cells from metastases. Many nuclear features of primary tumors and metastases correlated with the TNM stage. A specific pattern of correlation between well-established prognostic indicators and nuclear features of samples from primary tumors and those from neck metastases was observed. Image cytometric nuclear features represent a promising candidate marker for recognition of biologically different tumor subgroups

  12. Validation of flow cytometric analysis of platelet function in patients with a suspected platelet function defect.

    Science.gov (United States)

    van Asten, I; Schutgens, R E G; Baaij, M; Zandstra, J; Roest, M; Pasterkamp, G; Huisman, A; Korporaal, S J A; Urbanus, R T

    2018-01-16

    Essentials The diagnosis of mild platelet function disorders (PFDs) is challenging. Validation of flow cytometric testing in patients with suspected PFDs is required. Flow cytometry has added value to light transmission aggregometry (LTA) in diagnosis of PFDs. There is fair agreement in diagnosing PFDs between LTA and flow cytometry. Background Light transmission aggregometry (LTA) is the most commonly used test for the diagnosis of platelet function disorders (PFDs), but has moderate sensitivity for mild PFDs. Flow cytometry has been recommended for additional diagnostics of PFDs but is not yet standardized as a diagnostic test. We developed a standardized protocol for flow cytometric analysis of platelet function that measures fibrinogen binding and P-selectin expression as platelet activation markers in response to agonist stimulation. Objectives To determine the additional value of flow cytometric platelet function testing to standard LTA screening in a cross-sectional cohort of patients with a suspected PFD. Methods Platelet function was assessed with flow cytometry and LTA in 107 patients suspected of a PFD in whom von Willebrand disease and coagulation factor deficiencies were excluded. Both tests were compared in terms of agreement and discriminative ability for diagnosing patients with PFDs. Results Out of 107 patients, 51 patients had an elevated bleeding score; 62.7% of the patients had abnormal platelet function measured with flow cytometry and 54.2% of the patients were abnormal based on LTA. There was fair agreement between LTA and flow cytometry (κ = 0.32). The discriminative ability of flow cytometric analysis in patients with an elevated bleeding score was good (AUC 0.82, 0.74-0.90), but moderate for LTA (AUC 0.70, 0.60-0.80). Both tests combined had a better discriminative ability (AUC 0.87, 0.80-0.94). Conclusion Flow cytometric analysis of platelet function has added value in diagnostics of PFDs in patients with unexplained bleeding tendency.

  13. Flow cytometric immunophenotyping of adult T-cell leukemia/lymphoma using CD3 gating.

    Science.gov (United States)

    Yokote, Taiji; Akioka, Toshikazu; Oka, Satoko; Hara, Satoshi; Kobayashi, Kichinosuke; Nakajima, Hideto; Yamano, Takeshi; Ikemoto, Toshiyuki; Shimizu, Akira; Tsuji, Motomu; Hanafusa, Toshiaki

    2005-08-01

    Adult T-cell leukemia/lymphoma (ATLL) is a lymphoproliferative neoplasm of helper T lymphocytes caused by human T-cell leukemia virus type-1 (HTLV-1). The disease was first described in Kyushu, in southwestern Japan, and most frequently occurs in endemic areas, such as Japan, the Caribbean basin, West Africa, Brazil, and northern Iran. ATLL is essentially a disease of adults, characterized clinically by generalized lymphadenopathy, hepatosplenomegaly, skin lesions, and hypercalcemia. The prognosis of most patients is quite poor, with a median survival time of only 13 months, even if multiagent combination chemotherapy is given. In the present study, flow cytometric immunophenotyping with CD3 gating was performed on 30 samples from 26 patients who had been given a diagnosis of ATLL. The records of these patients also were reviewed retrospectively. In 14 of the 30 samples, an abnormal CD3(low) T-cell population was distinguishable from the normal T-cell populations by flow cytometric analysis. Herein we report a novel strategy for flow cytometric immunophenotyping of ATLL facilitated by CD3(low) gating.

  14. Single-Cell Phosphospecific Flow Cytometric Analysis of Canine and Murine Adipose-Derived Stem Cells

    Directory of Open Access Journals (Sweden)

    Harumichi Itoh

    2017-01-01

    Full Text Available This study aimed to demonstrate single-cell phosphospecific flow cytometric analysis of canine and murine adipose-derived stem/stromal cells (ADSCs. ADSCs were obtained from clinically healthy laboratory beagles and C57BL/6 mice. Cell differentiation into adipocytes, osteocytes, and chondrocytes was observed for the cultured canine ADSCs (cADSCs and murine ADSCs (mADSCs to determine their multipotency. We also performed single-cell phosphospecific flow cytometric analysis related to cell differentiation and stemness. Cultured cADSCs and mADSCs exhibited the potential to differentiate into adipocytes, osteocytes, and chondrocytes. In addition, single-cell phosphospecific flow cytometric analysis revealed similar β-catenin and Akt phosphorylation between mADSCs and cADSCs. On the other hand, it showed the phosphorylation of different Stat proteins. It was determined that cADSCs and mADSCs show the potential to differentiate into adipocytes, osteocytes, and chondrocytes. Furthermore, a difference in protein phosphorylation between undifferentiated cADSCs and mADSCs was identified.

  15. Multiparametric flow cytometric analysis of estrogen receptor: a ...

    African Journals Online (AJOL)

    Precise prognostication of breast cancer based on immunohistochemical features is a challenging assay. Thus, there is a need for more sophisticated prognostic determinants. This work aims to investigate the sensitivity of flow cytometry for the accurate evaluation of steroid receptor positive, tumor cells in formalin-fixed ...

  16. Flow cytometric chromosome sorting in plants: The next generation

    Czech Academy of Sciences Publication Activity Database

    Vrána, Jan; Šimková, Hana; Kubaláková, Marie; Čihalíková, Jarmila; Doležel, Jaroslav

    2012-01-01

    Roč. 57, č. 3 (2012), s. 331-337 ISSN 1046-2023 R&D Projects: GA ČR GAP501/10/1740 Grant - others:GA MŠk(CZ) ED0007/01/01 Program:ED Institutional research plan: CEZ:AV0Z50380511 Keywords : Chromosome sorting * Flow cytometry * Fluorescence in situ hybridization Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.641, year: 2012

  17. Ultrastructural and flow cytometric analyses of lipid accumulation in microalgae

    Energy Technology Data Exchange (ETDEWEB)

    Solomon, J.A.; Hand, R.E. Jr.; Mann, R.C.

    1986-12-01

    Lipid accumulation in three species of microalgae was investigated with flow cytometry (FCM) and transmission electron microscopy (TEM). Previous studies using batch cultures of a algae have led to the assumption that lipid accumulation in microalgae is a gradual process requiring at least several days for completion. However, FCM reveals, through changes in the chlorophyll:lipid ratio, that the time span required for individual cells to change metabolic state is short. Simultaneous FCM measurements of chlorophyll and nile red (neutral lipid) fluorescence in individual cells of nitrogen-deficient Isochrysis populations revealed a bimodal population distribution as one stage in the lipid accumulation process. The fact that two discrete populations exist, with few cells in an intermediate stage, suggests rapid response to a liqid trigger. Interpretations of light and electron microscopic observations are consistent with this hypothesis. The time required for an entire population to achieve maximum lipid content is considerably longer than that required for a single cell, due to the variation in response time among cells. In this study high lipid cultures were sometimes obtained by using FCM to separate high lipid cells from the remainder of the population. FCM holds much promise for strain enhancement but considerable developmental work, directed at providing more consistent results, remains to be done. 8 refs., 35 figs.

  18. Flow cytometric analysis of microbial contamination in food industry technological lines--initial study.

    Science.gov (United States)

    Józwa, Wojciech; Czaczyk, Katarzyna

    2012-04-02

    Flow cytometry constitutes an alternative for traditional methods of microorganisms identification and analysis, including methods requiring cultivation step. It enables the detection of pathogens and other microorganisms contaminants without the need to culture microbial cells meaning that the sample (water, waste or food e.g. milk, wine, beer) may be analysed directly. This leads to a significant reduction of time required for analysis allowing monitoring of production processes and immediate reaction in case of contamination or any disruption occurs. Apart from the analysis of raw materials or products on different stages of manufacturing process, the flow cytometry seems to constitute an ideal tool for the assessment of microbial contamination on the surface of technological lines. In the present work samples comprising smears from 3 different surfaces of technological lines from fruit and vegetable processing company from Greater Poland were analysed directly with flow cytometer. The measured parameters were forward and side scatter of laser light signals allowing the estimation of microbial cell contents in each sample. Flow cytometric analysis of the surface of food industry production lines enable the preliminary evaluation of microbial contamination within few minutes from the moment of sample arrival without the need of sample pretreatment. The presented method of fl ow cytometric initial evaluation of microbial state of food industry technological lines demonstrated its potential for developing a robust, routine method for the rapid and labor-saving detection of microbial contamination in food industry.

  19. Flow cytometric of reticulocytes quantification: radio-induction medullary aplasia application

    International Nuclear Information System (INIS)

    Dubner, D.; Perez, M.; Gisone, P.

    1996-01-01

    Flow cytometric reticulocyte quantification was assayed in ten patients undergoing bone marrow transplantation (BMT) with previous conditioning by chemotherapy and total body irradiation. A reticulocyte maturity index (RMI) was determined taking into account the RNA content. With the aim of testing the utility of RMI as an early predictor of functional recovery in marrow aplasia, other hematological indicators as neutrophils count were comparatively evaluated. Mean time elapsed between BMT and engraftment evidence by RMI was 17,6 days. In six patients the RMI was the earliest indicator of functional recovery. The applicability of this assay in the pursuit of radioinduced bone marrow aplasia is discussed. (authors). 4 refs., 4 figs., 2 tabs

  20. Karyological and flow cytometric evidence of triploid specimens in Bufo viridis (Amphibia Anura

    Directory of Open Access Journals (Sweden)

    D Cavallo

    2010-01-01

    Full Text Available Karyological and flow cytometric (FCM analyses were performed on a group of 14 green toads of the Bufo viridis species from seven Eurasian populations. Both approaches gave concordant results concerning the DNA ploidy level. All the populations examined were represented exclusively by diploid or tetraploid specimens, except one, where triploids were found. Results evidenced an interpopulation variability in DNA content against the same ploidy level, as well as an unusually high number of triploids in a particular reproductive place. The origin of polyploidy and the presence and persistence of a high number of triploids in a particular population are discussed.

  1. A flow cytometric assay technology based on quantum dots-encoded beads

    International Nuclear Information System (INIS)

    Wang Haiqiao; Liu Tiancai; Cao Yuancheng; Huang Zhenli; Wang Jianhao; Li Xiuqing; Zhao Yuandi

    2006-01-01

    A flow cytometric detecting technology based on quantum dots (QDs)-encoded beads has been described. Using this technology, several QDs-encoded beads with different code were identified effectively, and the target molecule (DNA sequence) in solution was also detected accurately by coupling to its complementary sequence probed on QDs-encoded beads through DNA hybridization assay. The resolution of this technology for encoded beads is resulted from two longer wavelength fluorescence identification signals (yellow and red fluorescent signals of QDs), and the third shorter wavelength fluorescence signal (green reporting signal of fluorescein isothiocyanate (FITC)) for the determination of reaction between probe and target. In experiment, because of QDs' unique optical character, only one excitation light source was needed to excite the QDs and probe dye FITC synchronously comparing with other flow cytometric assay technology. The results show that this technology has present excellent repeatability and good accuracy. It will become a promising multiple assay platform in various application fields after further improvement

  2. Detection of nickel and palladium contact hypersensitivity by a flow cytometric lymphocyte proliferation test.

    Science.gov (United States)

    Spoerri, I; Scherer, K; Michel, S; Link, S; Bircher, A J; Heijnen, I A F M

    2015-03-01

    We established a flow cytometric lymphocyte proliferation test (LPT) for the detection of nickel (Ni) and palladium (Pd) sensitization. Eighty-one consecutive patients with an indication for patch test (PT) were tested by LPT with Ni (NiSO4 ) and Pd (Na2 PdCl4 and PdCl2 ) salts. The imprecision of the LPT was low (coefficient of variation 7.2%). Using PT as a diagnostic reference, the sensitivity and specificity of LPT were 74.4% and 80% for NiSO4 , 74.4% and 78.3% for Na2 PdCl4 , and 57.2% and 85.4% for PdCl2 , respectively. For both Ni and Pd, the likelihood ratio for a positive PT markedly increased with increasing LPT value. With medical history as a reference, the sensitivity and specificity were 40.6% and 82.1% for LPT and 59.4% and 89.7% for PT, respectively. Combination of LPT and PT resulted in a higher specificity of 95%, albeit lower sensitivity of 34.4%. In conclusion, flow cytometric LPT represents a reliable and useful method for the detection of Ni and Pd sensitization. LPT values correlate with PT results and, when used in combination with PT, increase test specificity. © 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  3. A flow cytometric protocol for titering recombinant adenoviral vectors containing the green fluorescent protein.

    Science.gov (United States)

    Hitt, D C; Booth, J L; Dandapani, V; Pennington, L R; Gimble, J M; Metcalf, J

    2000-03-01

    As the use of adenoviral vectors in gene therapy protocols increases, there is a corresponding need for rapid, accurate, and reproducible titer methods. Multiple methods currently exist for determining titers of recombinant adenoviral vector, including optical absorbance, electron microscopy, fluorescent focus assay, and the "gold standard" plaque assay. This paper introduces a novel flow cytometric method for direct titer determination that relies on the expression of the green fluorescent protein (GFP), a tracking marker incorporated into several adenoviral vectors. This approach was compared to the plaque assay using 10(-4)- to 10(-6)-fold dilutions of a cesium-chloride-purified, GFP expressing adenovirus (AdEasy + GFP + GAL). The two approaches yielded similar titers: 3.25 +/- 1.85 x 10(9) PFU/mL versus 3.46 +/- 0.76 x 10(9) green fluorescent units/(gfu/mL). The flow cytometric method is complete within 24 h in contrast to the 7 x 10 days required by the plaque assay. These results indicate that the GFU/mL is an alternative functional titer method for fluorescent-tagged adenoviral vectors.

  4. Multiplex competitive microbead-based flow cytometric immunoassay using quantum dot fluorescent labels

    International Nuclear Information System (INIS)

    Yu, Hye-Weon; Kim, In S.; Niessner, Reinhard; Knopp, Dietmar

    2012-01-01

    Highlights: ► First time, duplex competitive bead-based flow cytometric immunoassay was developed using ODs. ► Antibody-coated QD detection probes and antigen-immobilized microspheres were synthesized. ► The two model target analytes were low molecular weight compounds of microbial and chemical origin. ► The determination of different water types was possible after simple filtration of samples. - Abstract: In answer to the ever-increasing need to perform the simultaneous analysis of environmental hazards, microcarrier-based multiplex technologies show great promise. Further integration with biofunctionalized quantum dots (QDs) creates new opportunities to extend the capabilities of multicolor flow cytometry with their unique fluorescence properties. Here, we have developed a competitive microbead-based flow cytometric immunoassay using QDs fluorescent labels for simultaneous detection of two analytes, bringing the benefits of sensitive, rapid and easy-of-manipulation analytical tool for environmental contaminants. As model target compounds, the cyanobacterial toxin microcystin-LR and the polycyclic aromatic hydrocarbon compound benzo[a]pyrene were selected. The assay was carried out in two steps: the competitive immunological reaction of multiple targets using their exclusive sensing elements of QD/antibody detection probes and antigen-coated microsphere, and the subsequent flow cytometric analysis. The fluorescence of the QD-encoded microsphere was thus found to be inversely proportional to target analyte concentration. Under optimized conditions, the proposed assay performed well within 30 min for the identification and quantitative analysis of the two environmental contaminants. For microcystin-LR and benzo[a]pyrene, dose–response curves with IC 50 values of 5 μg L −1 and 1.1 μg L −1 and dynamic ranges of 0.52–30 μg L −1 and 0.13–10 μg L −1 were obtained, respectively. Recovery was 92.6–106.5% for 5 types of water samples like bottled

  5. Flow Cytometric Analysis of T, B, and NK Cells Antigens in Patients with Mycosis Fungoides

    Directory of Open Access Journals (Sweden)

    Serkan Yazıcı

    2015-01-01

    Full Text Available We retrospectively analyzed the clinicopathological correlation and prognostic value of cell surface antigens expressed by peripheral blood mononuclear cells in patients with mycosis fungoides (MF. 121 consecutive MF patients were included in this study. All patients had peripheral blood flow cytometry as part of their first visit. TNMB and histopathological staging of the cases were retrospectively performed in accordance with International Society for Cutaneous Lymphomas/European Organization of Research and Treatment of Cancer (ISCL/EORTC criteria at the time of flow cytometry sampling. To determine prognostic value of cell surface antigens, cases were divided into two groups as stable and progressive disease. 17 flow cytometric analyses of 17 parapsoriasis (PP and 11 analyses of 11 benign erythrodermic patients were included as control groups. Fluorescent labeled monoclonal antibodies were used to detect cell surface antigens: T cells (CD3+, CD4+, CD8+, TCRαβ+, TCRγδ+, CD7+, CD4+CD7+, CD4+CD7−, and CD71+, B cells (HLA-DR+, CD19+, and HLA-DR+CD19+, NKT cells (CD3+CD16+CD56+, and NK cells (CD3−CD16+CD56+. The mean value of all cell surface antigens was not statistically significant between parapsoriasis and MF groups. Along with an increase in cases of MF stage statistically significant difference was found between the mean values of cell surface antigens. Flow cytometric analysis of peripheral blood cell surface antigens in patients with mycosis fungoides may contribute to predicting disease stage and progression.

  6. Bivariate flow cytometric analysis and sorting of different types of maize starch grains.

    Science.gov (United States)

    Zhang, Xudong; Feng, Jiaojiao; Wang, Heng; Zhu, Jianchu; Zhong, Yuyue; Liu, Linsan; Xu, Shutu; Zhang, Renhe; Zhang, Xinghua; Xue, Jiquan; Guo, Dongwei

    2018-02-01

    Particle-size distribution, granular structure, and composition significantly affect the physicochemical properties, rheological properties, and nutritional function of starch. Flow cytometry and flow sorting are widely considered convenient and efficient ways of classifying and separating natural biological particles or other substances into subpopulations, respectively, based on the differential response of each component to stimulation by a light beam; the results allow for the correlation analysis of parameters. In this study, different types of starches isolated from waxy maize, sweet maize, high-amylose maize, pop maize, and normal maize were initially classified into various subgroups by flow cytometer and then collected through flow sorting to observe their morphology and particle-size distribution. The results showed that a 0.25% Gelzan solution served as an optimal reagent for keeping individual starch particles homogeneously dispersed in suspension for a relatively long time. The bivariate flow cytometric population distributions indicated that the starches of normal maize, sweet maize, and pop maize were divided into two subgroups, whereas high-amylose maize starch had only one subgroup. Waxy maize starch, conversely, showed three subpopulations. The subgroups sorted by flow cytometer were determined and verified in terms of morphology and granule size by scanning electron microscopy and laser particle distribution analyzer. Results showed that flow cytometry can be regarded as a novel method for classifying and sorting starch granules. © 2017 International Society for Advancement of Cytometry. © 2017 International Society for Advancement of Cytometry.

  7. Improved flow cytometric assessment reveals distinct microvesicle (cell-derived microparticle signatures in joint diseases.

    Directory of Open Access Journals (Sweden)

    Bence György

    Full Text Available INTRODUCTION: Microvesicles (MVs, earlier referred to as microparticles, represent a major type of extracellular vesicles currently considered as novel biomarkers in various clinical settings such as autoimmune disorders. However, the analysis of MVs in body fluids has not been fully standardized yet, and there are numerous pitfalls that hinder the correct assessment of these structures. METHODS: In this study, we analyzed synovial fluid (SF samples of patients with osteoarthritis (OA, rheumatoid arthritis (RA and juvenile idiopathic arthritis (JIA. To assess factors that may confound MV detection in joint diseases, we used electron microscopy (EM, Nanoparticle Tracking Analysis (NTA and mass spectrometry (MS. For flow cytometry, a method commonly used for phenotyping and enumeration of MVs, we combined recent advances in the field, and used a novel approach of differential detergent lysis for the exclusion of MV-mimicking non-vesicular signals. RESULTS: EM and NTA showed that substantial amounts of particles other than MVs were present in SF samples. Beyond known MV-associated proteins, MS analysis also revealed abundant plasma- and immune complex-related proteins in MV preparations. Applying improved flow cytometric analysis, we demonstrate for the first time that CD3(+ and CD8(+ T-cell derived SF MVs are highly elevated in patients with RA compared to OA patients (p=0.027 and p=0.009, respectively, after Bonferroni corrections. In JIA, we identified reduced numbers of B cell-derived MVs (p=0.009, after Bonferroni correction. CONCLUSIONS: Our results suggest that improved flow cytometric assessment of MVs facilitates the detection of previously unrecognized disease-associated vesicular signatures.

  8. Cell cycle flow cytometric analysis in the diagnosis and management of colorectal carcinoma.

    Science.gov (United States)

    Sampedro, A; Salas-Bustamante, A; López-Artimez, M; García-Muñíz, J L; Urdiales, G

    1999-08-01

    To establish prognostic models and protocols for individualized management in colorectal carcinoma patients based on both clinical and DNA flow cytometric parameters. Prospective study of 88 colon carcinoma patients with a minimum follow-up of 12 months, operated on with the intent to cure and not treated with radiotherapy or chemotherapy. All the cases were subjected to a clinical evaluation: age, sex, tumor localization and size, histologic grade, tumor stage, disease-free interval, survival and flow cytometric study (ploidy, DNA index and S-phase fraction [SPF]). From the total of 88 neoplasms studied, 56 (63.6%) were from males and 32 (36.4%) from females; 30 (34%) were located in the right side of the colon, 7 (8%) in the transverse colon and 51 (58%) in the left side of the colon. Eleven (12.5%) were stage I, 52 (59.1%) stage II and 25 (28%) stage III. Forty-two (47.7%) were diploid and 46 (52.3%) aneuploid. The S-phase mean was 14.6% (12% for diploids and 16.9% for aneuploids). During the follow-up period, 26.1% of diploid tumors recurred, whereas aneuploid tumors recurred in 36.9% (P < .05). SPF from diploid and aneuploid tumors was analyzed separately. Regarding relapse-free interval, the behavior of diploid tumors with a high SPF was similar to that of aneuploid ones. Two kinetic profiles were established, favorable (diploid tumors with low S phase) and unfavorable (diploid with high S phase and all aneuploid tumors), that had significant prognostic value for progression and survival and that allowed identification of patients at high risk of recurrence. We formulated a prognostic index according to SPF and tumor stage that has discriminatory capacity for biologic behavior in colorectal tumors.

  9. A new one-platform flow cytometric method for residual cell counting in platelet concentrates.

    Science.gov (United States)

    Schmidt, Michael; Spengler, Hans-Peter; Lambrecht, Bernd; Hourfar, Michael K; Seifried, Erhard; Tonn, Torsten

    2009-12-01

    According to German regulations and guidelines, residual red blood cells (rRBCs) and residual white blood cells (rWBCs) must number fewer than 3 x 10(9) cells/unit and 1 x 10(6) cells/unit in platelet concentrates (PCs), respectively. Due to low levels of residual cells in final products, there is still a need for fast, reliable, and sensitive methods of automated detection of these cell types. In Part A, 21 PCs were spiked with predetermined numbers of red blood cells (RBCs) and white blood cells (WBCs). The linearity, precision, and accuracy of the BD Thrombo Count assay (BD Biosciences Europe) were tested and validated according to international guidelines. Finally in Part B, 100 PCs prepared from pooled buffy coats were tested by the BD Thrombo Count assay and compared with other methods, including Nageotte (rWBCs) and Neubauer (rRBCs) counting chambers and the flow cytometric BD LeucoCOUNT (Becton Dickinson) assay (rWBCs). The unspecific background of blank PC samples was fewer than 0.02 cells/microL for WBCs and fewer than 34 cells/microL for RBCs (mean, 21). Linear regression and precision analyses of spiked PC samples were determined for both WBCs (r(2) = 0.992; range, 0.6-6.0 WBCs/microL) and RBCs (r(2) = 0.999; 800-8000 RBCs/microL). No carryover of cells or drift in results was detected in the automated sample acquisition mode. Analysis according to statistical methods of Bland and Altman demonstrated a high correlation between BD Thrombo Count and the Neubauer manual counting chamber. This novel flow cytometric test is a quick and reliable single-tube assay that has been demonstrated as a potential alternative for the existing manual microscopic counting procedures that are both time-consuming and laborious.

  10. Biotinylation of interleukin-2 (IL-2) for flow cytometric analysis of IL-2 receptor expression. Comparison of different methods

    NARCIS (Netherlands)

    M.O. de Jong (Marg); H. Rozemuller (Henk); J.G.J. Bauman (J. G J); J.W.M. Visser (Jan)

    1995-01-01

    textabstractThe main prerequisites for the use of biotinylated ligands to study the expression of growth factor receptors on heterogeneous cell populations, such as peripheral blood or bone marrow, by flow cytometric methods, are that the biotinylated ligand retains its binding ability and that

  11. Clonal evolution demonstrated by flow cytometric DNA analysis of a human colonic carcinoma grown in nude mice

    DEFF Research Database (Denmark)

    Vindeløv, L L; Spang-Thomsen, M; Visfeldt, J

    1982-01-01

    A spontaneous change in DNA content of a human colonic carcinoma grown in nude mice was observed fortuitously. The tumor initially had a G1 cell DNA content of 1.3 times that of normal cells. Flow cytometric DNA analysis showed in transplant generation 56 the appearance of a new subpopulation which...

  12. A novel flow cytometric assay for measurement of In Vivo pulmonary neutrophil phagocytosis

    Directory of Open Access Journals (Sweden)

    Gentry-Nielsen Martha J

    2006-07-01

    Full Text Available Abstract Background Phagocytosis assays are traditionally performed in vitro using polymorphonuclear leukocytes (PMNs isolated from peripheral blood or the peritoneum and heat-killed, pre-opsonized organisms. These assays may not adequately mimic the environment within the infected lung. Our laboratory therefore has developed a flow cytometric in vivo phagocytosis assay that enables quantification of PMN phagocytosis of viable bacteria within the lungs of rats. In these studies, rats are injected transtracheally with lipopolysaccharide (LPS to recruit PMNs to their lungs. They are then infected with live 5(-and 6 carboxyfluorescein diacetate succinimidyl ester (CFDA/SE labeled type 3 Streptococcus pneumoniae. Bronchoalveolar lavage is performed and resident alveolar macrophages and recruited PMNs are labeled with monoclonal antibodies specific for surface epitopes on each cell type. Three color flow cytometry is utilized to identify the cell types, quantify recruitment, and determine uptake of the labeled bacteria. Results The viability of the alveolar macrophages and PMNs isolated from the lavage fluid was >95%. The values of the percentage of PMNs in the lavage fluid as well as the percentage of PMNs associated with CFSE-labeled S. pneumoniae as measured through flow cytometry showed a high degree of correlation with the results from manual counting of cytospin slides. Conclusion This assay is suitable for measuring bacterial uptake within the infected lung. It can be adapted for use with other organisms and/or animal model systems.

  13. Flow cytometric analysis of the granulocyte respiratory burst: a comparison study of fluorescent probes.

    Science.gov (United States)

    Vowells, S J; Sekhsaria, S; Malech, H L; Shalit, M; Fleisher, T A

    1995-01-13

    Chronic granulomatous disease (CGD) is a rare recessive disorder caused by defects in the NADPH oxidase enzyme complex of phagocytes (neutrophils, eosinophils and monocytes). CGD phagocytes fail to produce superoxide and other reactive oxygen species following cell activation (Malech, 1993). The products of oxidase activation can be measured in individual cells by flow cytometry using specific fluorescent probes that increase fluorescence upon oxidation (Trinkle et al., 1987). This approach can be used to confirm a diagnosis of CGD, and to detect the normal/abnormal phagocyte mixture that characterizes the X-linked CGD carrier state. Three fluorescent probes have been described as useful for this purpose: 2'7'-dichlorofluorescin diacetate (DCF) (Bass et al., 1983), 5,6-carboxy-2'7'-dichlorofluorescein diacetate, bis(acetoxymethyl) ester (C-DCF) (Hockenbery et al., 1993) and dihydrorhodamine 123 (DHR) (Rothe et al., 1988; Kinsey et al., 1987). A direct comparison between these three probes has not been reported. In this study we performed a direct comparison between these three probes, evaluating their ability in flow cytometric analysis to maximize fluorescent separation between activated CGD patient and normal granulocytes. Using a whole blood technique with phorbol myristate acetate (PMA) as an activator, it was found that DHR loaded normal granulocytes had a fluorescence intensity which, upon activation, was 48-fold higher than that of C-DCF loaded granulocytes and seven-fold higher than DCF loaded granulocytes (P < 0.001). Use of sodium azide to decrease the catabolism of H2O2 enhanced the fluorescence of DCF by 140%, C-DCF by 45% and DHR by 25%, suggesting that DCF is primarily sensitive to H2O2. DCF and DHR were then evaluated for sensitivity in the detection of small percentages of normal cells in a CGD/normal granulocyte mixture. Normal sub-populations as small as 0.1% could clearly be distinguished using DHR, while DCF was insensitive at this level. Based

  14. Novel calibration method for flow cytometric fluorescence resonance energy transfer measurements between visible fluorescent proteins.

    Science.gov (United States)

    Nagy, Peter; Bene, László; Hyun, William C; Vereb, György; Braun, Manuela; Antz, Christof; Paysan, Jacques; Damjanovich, Sándor; Park, John W; Szöllősi, János

    2005-10-01

    The combination of fluorescence resonance energy transfer (FRET) and flow cytometry offers a statistically firm approach to study protein associations. Fusing green fluorescent protein (GFP) to a studied protein usually does not disturb the normal function of a protein, but quantitation of FRET efficiency calculated between GFP derivatives poses a problem in flow cytometry. We generated chimeras in which cyan fluorescent protein (CFP) was separated by amino acid linkers of different sizes from yellow fluorescent protein (YFP) and used them to calibrate the cell-by-cell flow cytometric FRET measurements carried out on two different dual-laser flow cytometers. Then, CFP-Kip1 was coexpressed in yeast cells with YFP and cyclin-dependent kinase-2 (Cdk2) and served as a positive control for FRET measurements, and CFP-Kip1 coexpressed with a random peptide fused to YFP was the negative control. We measured donor, direct, and sensitized acceptor fluorescence intensities and developed a novel way to calculate a factor (alpha) that characterized the fluorescence intensity of acceptor molecules relative to the same number of excited donor molecules, which is essential for quantifying FRET efficiency. This was achieved by calculating FRET efficiency in two different ways and minimizing the squared difference between the two results by changing alpha. Our method reliably detected the association of Cdk2 with its inhibitor, Kip1, whereas the nonspecific FRET efficiency between Cdk2 and a random peptide was negligible. We identified and sorted subpopulations of yeast cells showing interaction between the studied proteins. We have described a straightforward novel calibration method to accurately quantitate FRET efficiency between GFP derivatives in flow cytometry.

  15. A flow cytometric method for characterization of circulating cell-derived microparticles in plasma

    Directory of Open Access Journals (Sweden)

    Morten Hjuler Nielsen

    2014-02-01

    Full Text Available Background and aim: Previous studies on circulating microparticles (MPs indicate that the majority of MPs are of a size below the detection limit of most standard flow cytometers. The objective of the present study was to establish a method to analyze MP subpopulations above the threshold of detection of a new generation BD FACSAria™ III digital flow cytometer. Methods: We analyzed MP subpopulations in plasma from 24 healthy individuals (9 males and 15 females. MPs were identified according to their size (<1.0-µm, by Lactadherin-FITC labelling, and by exposure of cell-specific markers. The sensitivity of the flow cytometer was tested against that of a previous-generation instrument FC500. Reproducibility of the FACSAria and our set-up was investigated, and the percentage of phosphatidylserine (PS exposing MPs binding Lactadherin was determined. Results: By using a flow cytometric approach we identified and quantitated MPs derived from platelets, monocytes, erythrocytes and endothelial cells. In addition, levels of tissue factor-positive MPs were determined. The FACSAria demonstrated improved sensitivity and increased MP detection range compared to the FC500 instrument. The reproducibility of PS+PMP and PS+MP measurements was 11.7 and 23.2%, respectively. When expressed as a percentage of total MPs, the PS-positive MP population represented 15.1±5.5%, and PS-positive MPs were significantly increased in men. Conclusion: We have established a method to measure MPs above the detection limit of a new generation flow cytometer and derived from a number of cell-types in a healthy population of men and women.

  16. Laser-based flow cytometric analysis of genotoxicity of humans exposed to ionizing radiation during the Chernobyl accident

    Science.gov (United States)

    Jensen, Ronald H.; Bigbee, William L.; Langlois, Richard G.; Grant, Stephen G.; Pleshanov, Pavel G.; Chirkov, Andre A.; Pilinskaya, Maria A.

    1991-05-01

    An analytical technique has been developed that allows laser-based flow cytometric measurement of the frequency of red blood cells that have lost allele-specific expression of a cell surface antigen due to genetic toxicity in bone marrow precursor cells. Previous studies demonstrated a correlation of such effects with the exposure of each individual to mutagenic phenomena, such as ionizing radiation, and the effects can persist for the lifetime of each individual. During the emergency response to the nuclear power plant accidert at Chemobyl, Ukraine, USSR, a number of people were exposed to whole body doses of ioniing radiation. Some of these individuals were tested with this laser-based assay and found to express a dose-dependent increase in the frequency of variant red blood cells that appears to be a persistent biological effect. This effect is similar to that which was previously observed in individuals who were exposed to ionizing radiation at Hiroshima in 1945 because of the A-bomb explosion. All data indicate that this assay might well be used as a biodosimeter to estimate radiation dose and also as an element to be used for estimating the risk of each individual to develop cancer due to radiation exposure.

  17. Dissociation of mono- and co-culture spheroids into single cells for subsequent flow cytometric analysis.

    Science.gov (United States)

    Grässer, Ute; Bubel, Monika; Sossong, Daniela; Oberringer, Martin; Pohlemann, Tim; Metzger, Wolfgang

    2018-03-01

    Spheroids are considered to reflect the natural organization of cells better than 2D cell cultures, but their analysis by flow cytometry requires dissociation into single cells. We established protocols for dissociation of mono- and co-culture spheroids consisting of human fibroblasts and human endothelial cells. Cell recovery rate and viability after dissociation were evaluated with hemocytometer and by flow cytometry. The diameter of cells and the amount of cell aggregates were quantified by Casy ® -technology and the cellular composition was analyzed by flow cytometry. Optimal dissociation conditions with low cell aggregation were determined by size, cultivation time and cellular composition of the spheroids. Smaller spheroids (10,000 cells) could be dissociated with Accutase ® , whereas larger spheroids (50,000 cells) required more stringent dissociation conditions. The size of the cells decreased with increasing cultivation time. Cell recovery rate was dependent upon cellular composition and spheroid size. The highest cell recovery rate was found for co-culture spheroids. The highest cell viability was detected for dissociated fibroblast spheroids. A quantitative analysis of the cellular composition of dissociated co-culture spheroids was possible. Spheroids can be successfully dissociated into singular cells for subsequent flow cytometric analysis. Dissociation conditions as well as cell recovery rate and cell viability depend on size, cultivation time and cellular composition of the spheroids. The observed decrease in cell size in spheroids over time might be responsible for the well-known time-dependent decrease in spheroid size. Copyright © 2017 Elsevier GmbH. All rights reserved.

  18. Early changes in flow cytometric DNA profiles induced by californium-252 neutron brachytherapy in squamocellular carcinomas of the uterine cervix.

    Science.gov (United States)

    Tacev, T; Zaloudík, J; Janáková, L; Vagunda, V

    1998-01-01

    Ninety-five squamocellular carcinomas of the uterine cervix, clinical Stages II and III, were treated by either four schedules combining 252-californium neutron-gamma-radiotherapy with different proportions of a neutron component (9, 6 and 3 Gy) or gamma-irradiation alone. Flow cytometric DNA profiles were obtainable in 72 cases before treatment and 56 cases were monitored for DNA content by flow cytometry (FCM) in weekly intervals by analysis of sequential microbiopsies for one month during and after radiotherapy. DNA aneuploidy was reduced from 40% (25/63) to 19% (9/47) one week within therapy in neutron-treated groups, but not after initial gamma-radiotherapy alone. Extinction of DNA aneuploid subpopulations occurred after neutron therapy in all remaining aneuploid tumors (9/9) during further monitoring, but only in 40% (2/5) of tumors after sole gamma-irradiation. In contrast, proliferation index by more than 50% was more often achieved in groups with a higher gamma-radiation component than after neutrons only. When all therapy-induced DNA flow cytometric events are taken together for evaluation of the effects of various radiotherapy schedules, it appears that the regimen with the maximal neutron dose may not be optimal for all tumors. It is hypothesized that the differences in the early flow cytometric DNA profiles may select the DNA aneuploid squamous cell uterine cervical carcinomas as candidates for combined neutron-brachytherapy, while highly proliferating DNA near-diploid tumors may profit more from treatment with a higher gamma-radiotherapy component. However, these early DNA flow cytometric findings need to be correlated with clinical course of the disease to validate this hypothesis, a process which will be completed at the end of the expected five-year clinical outcome in 2000.

  19. A novel rapid and reproducible flow cytometric method for optimization of transfection efficiency in cells.

    Science.gov (United States)

    Homann, Stefanie; Hofmann, Christian; Gorin, Aleksandr M; Nguyen, Huy Cong Xuan; Huynh, Diana; Hamid, Phillip; Maithel, Neil; Yacoubian, Vahe; Mu, Wenli; Kossyvakis, Athanasios; Sen Roy, Shubhendu; Yang, Otto Orlean; Kelesidis, Theodoros

    2017-01-01

    Transfection is one of the most frequently used techniques in molecular biology that is also applicable for gene therapy studies in humans. One of the biggest challenges to investigate the protein function and interaction in gene therapy studies is to have reliable monospecific detection reagents, particularly antibodies, for all human gene products. Thus, a reliable method that can optimize transfection efficiency based on not only expression of the target protein of interest but also the uptake of the nucleic acid plasmid, can be an important tool in molecular biology. Here, we present a simple, rapid and robust flow cytometric method that can be used as a tool to optimize transfection efficiency at the single cell level while overcoming limitations of prior established methods that quantify transfection efficiency. By using optimized ratios of transfection reagent and a nucleic acid (DNA or RNA) vector directly labeled with a fluorochrome, this method can be used as a tool to simultaneously quantify cellular toxicity of different transfection reagents, the amount of nucleic acid plasmid that cells have taken up during transfection as well as the amount of the encoded expressed protein. Finally, we demonstrate that this method is reproducible, can be standardized and can reliably and rapidly quantify transfection efficiency, reducing assay costs and increasing throughput while increasing data robustness.

  20. Fine needle aspirate flow cytometric phenotyping characterizes immunosuppressive nature of the mesothelioma microenvironment.

    Science.gov (United States)

    Lizotte, Patrick H; Jones, Robert E; Keogh, Lauren; Ivanova, Elena; Liu, Hongye; Awad, Mark M; Hammerman, Peter S; Gill, Ritu R; Richards, William G; Barbie, David A; Bass, Adam J; Bueno, Raphael; English, Jessie M; Bittinger, Mark; Wong, Kwok-Kin

    2016-08-19

    With the emergence of checkpoint blockade and other immunotherapeutic drugs, and the growing adoption of smaller, more flexible adaptive clinical trial designs, there is an unmet need to develop diagnostics that can rapidly immunophenotype patient tumors. The ability to longitudinally profile the tumor immune infiltrate in response to immunotherapy also presents a window of opportunity to illuminate mechanisms of resistance. We have developed a fine needle aspirate biopsy (FNA) platform to perform immune profiling on thoracic malignancies. Matching peripheral blood, bulk resected tumor, and FNA were analyzed from 13 mesothelioma patients. FNA samples yielded greater numbers of viable cells when compared to core needle biopsies. Cell numbers were adequate to perform flow cytometric analyses on T cell lineage, T cell activation and inhibitory receptor expression, and myeloid immunosuppressive checkpoint markers. FNA samples were representative of the tumor as a whole as assessed by head-to-head comparison to single cell suspensions of dissociated whole tumor. Parallel analysis of matched patient blood enabled us to establish quality assurance criteria to determine the accuracy of FNA procedures to sample tumor tissue. FNA biopsies provide a diagnostic to rapidly phenotype the tumor immune microenvironment that may be of great relevance to clinical trials.

  1. Flow cytometric analysis of lymphocyte proliferative responses to food allergens in dogs with food allergy.

    Science.gov (United States)

    Fujimura, Masato; Masuda, Kenichi; Hayashiya, Makio; Okayama, Taro

    2011-10-01

    Two different allergy tests, antigen-specific immunoglobulin E quantification (IgE test) and flow cytometric analysis of antigen-specific proliferation of peripheral lymphocytes (lymphocyte proliferation test), were performed to examine differences in allergic reactions to food allergens in dogs with food allergy (FA). Thirteen dogs were diagnosed as FA based on clinical findings and elimination diet trials. Seven dogs clinically diagnosed with canine atopic dermatitis (CAD) were used as a disease control group, and 5 healthy dogs were used as a negative control group. In the FA group, 19 and 33 allergen reactions were identified using the serum IgE test and the lymphocyte proliferation test, respectively. Likewise, in the CAD group, 12 and 6 allergen reactions and in the healthy dogs 3 and 0 allergen reactions were identified by each test, respectively. A significant difference was found between FA and healthy dogs in terms of positive allergen detection by the lymphocyte proliferation test, suggesting that the test can be useful to differentiate FA from healthy dogs but not from CAD. Both tests were repeated in 6 of the dogs with FA after a 1.5- to 5-month elimination diet trial. The IgE concentrations in 9 of 11 of the positive reactions decreased by 20-80%, whereas all the positive reactions in the lymphocyte proliferation test decreased to nearly zero (Pfood allergens may be involved in the pathogenesis of canine FA.

  2. A Flow Cytometric Analysis of Vitreous Inflammatory Cells in Patients with Proliferative Diabetic Retinopathy

    Directory of Open Access Journals (Sweden)

    Mojca Urbančič

    2013-01-01

    Full Text Available The purpose of this study was to investigate inflammatory cells in vitreous from patients with proliferative diabetic retinopathy (PDR using flow cytometric analysis. Twenty-eight patients with PDR requiring vitrectomy because of macular traction or tractional retinal detachment were enrolled in the study (n=28, and 6 patients with macular hole (MH formed the control group. Samples of vitreous and peripheral venous blood were obtained at the beginning of vitrectomy. T lymphocytes were found in vitreous from patients with PDR, and CD4/CD8 ratio was higher in vitreous (median 4.3 compared to blood (median 1.9; P=0.003. No B lymphocytes were detected in vitreous. The percentage of histiocytes/macrophages was significantly higher in vitreous (median 62.1 in comparison with blood (median 5.5; P<0.0001. No lymphocytes were detected in vitreous of the control group. There were more T lymphocytes in vitreous from patients with active PDR. No association between cells in the vitreous and visual acuity improvement after surgery was found. In conclusion, T lymphocytes are found in vitreous from patients with PDR and reflect the activity of PDR but do not seem to predict visual prognosis. Higher CD4/CD8 ratio in vitreous compared to blood from patients with PDR is consistent with local inflammatory response in PDR.

  3. Flow cytometric probing of mitochondrial function in equine peripheral blood mononuclear cells

    Directory of Open Access Journals (Sweden)

    Coignoul Freddy

    2007-09-01

    Full Text Available Abstract Background The morphopathological picture of a subset of equine myopathies is compatible with a primary mitochondrial disease, but functional confirmation in vivo is still pending. The cationic dye JC-1 exhibits potential-dependent accumulation in mitochondria that is detectable by a fluorescence shift from green to orange. As a consequence, mitochondrial membrane potential can be optically measured by the orange/green fluorescence intensity ratio. A flow cytometric standardized analytic procedure of the mitochondrial function of equine peripheral blood mononuclear cells is proposed along with a critical appraisal of the crucial questions of technical aspects, reproducibility, effect of time elapsed between blood sampling and laboratory processing and reference values. Results The JC-1-associated fluorescence orange and green values and their ratio were proved to be stable over time, independent of age and sex and hypersensitive to intoxication with a mitochondrial potential dissipator. Unless time elapsed between blood sampling and laboratory processing does not exceed 5 hours, the values retrieved remain stable. Reference values for clinically normal horses are given. Conclusion Whenever a quantitative measurement of mitochondrial function in a horse is desired, blood samples should be taken in sodium citrate tubes and kept at room temperature for a maximum of 5 hours before the laboratory procedure detailed here is started. The hope is that this new test may help in confirming, studying and preventing equine myopathies that are currently imputed to mitochondrial dysfunction.

  4. A Flow-Cytometric Gram-Staining Technique for Milk-Associated Bacteria

    Science.gov (United States)

    Holm, Claus; Jespersen, Lene

    2003-01-01

    A Gram-staining technique combining staining with two fluorescent stains, Oregon Green-conjugated wheat germ agglutinin (WGA) and hexidium iodide (HI) followed by flow-cytometric detection is described. WGA stains gram-positive bacteria while HI binds to the DNA of all bacteria after permeabilization by EDTA and incubation at 50°C for 15 min. For WGA to bind to gram-positive bacteria, a 3 M potassium chloride solution was found to give the highest fluorescence intensity. A total of 12 strains representing some of the predominant bacterial species in bulk tank milk and mixtures of these were stained and analyzed by flow cytometry. Overall, the staining method showed a clear differentiation between gram-positive and gram-negative bacterial populations. For stationary-stage cultures of seven gram-positive bacteria and five gram-negative bacteria, an average of 99% of the cells were correctly interpreted. The method was only slightly influenced by the growth phase of the bacteria or conditions such as freezing at −18°C for 24 h. For any of these conditions, an average of at least 95% of the cells were correctly interpreted. When stationary-stage cultures were stored at 5°C for 14 days, an average of 86% of the cells were correctly interpreted. The Gram-staining technique was applied to the flow cytometry analysis of bulk tank milk inoculated with Staphylococcus aureus and Escherichia coli. These results demonstrate that the technique is suitable for analyzing milk samples without precultivation. PMID:12732558

  5. Cholesterol sensitivity of detergent resistance: A rapid flow cytometric test for detecting constitutive or induced raft association of membrane proteins

    OpenAIRE

    Imre Gombos, Zsolt Bacsó, Cynthia Detre, Henrietta Nagy, Katalin Goda, Márton Andrásfalvy, Gábor Szabó, János Matkó; Bacsó Zsolt (1963-) (biofizikus); Goda Katalin (1969-) (biofizikus); Szabó Gábor (1953-) (biofizikus)

    2004-01-01

    Lipid rafts are cholesterol- and glycosphingolipid-rich microdomains in the cellular plasma membranes that play critical roles in compartmentalization (concentration, coupling, and isolation) of receptors and signal molecules. Therefore, detecting constitutive or induced raft associations of such proteins is of central interest in cell biology. This has mostly been done with time- and cell-consuming immunobiochemical techniques affected by several sources of artifacts. A flow cytometric analy...

  6. THE EFFECT OF LABELING INTENSITY, ESTIMATED BY REAL-TIME CONFOCAL LASER SCANNING MICROSCOPY, ON FLOW CYTOMETRIC APPEARANCE AND IDENTIFICATION OF IMMUNOCHEMICALLY LABELED MARINE DINOFLAGELLATES

    NARCIS (Netherlands)

    VRIELING, EG; DRAAIJER, A; VANZEIJL, WJM; PEPERZAK, L; GIESKES, WWC; VEENHUIS, M; Zeijl, Wilhelmus J.M. van

    Two different fluorescein isothiocyanate (FITC) conjugates were used to analyze the effect of labeling intensity on the flow cytometric appearance of marine dinoflagellates labeled with antibodies that specifically recognized the outer cell wall. Location of the labeling was revealed by

  7. Differentiation of HL-60 promyelocytic leukemia cells monitored by flow cytometric measurement of nitro blue tetrazolium (NBT) reduction.

    Science.gov (United States)

    Blair, O C; Carbone, R; Sartorelli, A C

    1985-01-01

    Reduction of nitro blue tetrazolium (NBT) to insoluble blue formazan granules occurs during the stimulus-induced respiratory burst of mature granulocytes and is routinely used as an indicator of the extent of granulocytic differentiation of HL-60 acute promyelocytic leukemia cells. In the present study, the differentiation of HL-60 leukemia cells induced by dimethylsulfoxide (DMSO) or retinoic acid was monitored by flow cytometric (FCM) measurement of forward and 90 degree light scatter of NBT treated cells. Two-parameter correlated analysis permitted a distinction between cells with increased forward and decreased 90 degree light scatter (NBT-), and cells with decreased forward and increased 90 degree light scatter (NBT+). Fixation of NBT treated cells with 1% paraformaldehyde facilitated flow cytometric analysis, and allowed differences in NBT reduction to be quantitated. DMSO-induced cells expressed an all-or-none reduction of NBT to formazan, compared with retinoic acid treated cells that exhibited a graded response. Three parameter flow cytometric analysis of HL-60 leukemia cells stained with propidium iodide in combination with NBT allowed the determination of the cell cycle distribution of NBT-treated cells.

  8. Flow cytometric analysis of viable bacteria in urine samples of febrile patients at the emergency department.

    Science.gov (United States)

    Tavenier, Anne H; de Boer, Foppie J; Moshaver, Bijan; van der Leur, Sjef J C M; Stegeman, Coen A; Groeneveld, Paul H P

    2017-08-16

    Fast and reliable diagnostics are important in febrile patients admitted to the emergency department. Current urine diagnostics are fast but moderately reliable or reliable but time consuming. Flow cytometry (FC) is a new promising technique in the diagnostics of complicated urinary tract infections by counting bacteria in urine samples. The aim of this study is to improve the FC method by counting only viable bacteria. Urine was obtained from 135 consecutive febrile patients at the emergency department. According to protocol regular diagnostic urine tests were performed. In addition, FC counting of viable and non-viable bacteria was executed after staining with thiazole orange and propidium iodide. All test results were compared to the results of urine culture (≥ 10 5 colony forming units/mL). At a cut-off value of 2.01 × 10 5 viable bacteria/mL the sensitivity was 100% and specificity was 78.4% (AUC-value 0.955 on ROC-curve). Spearman correlation test exhibited a higher correlation for flow cytometric counting of only viable bacteria than counting of all bacteria (0.59 vs. 0.37). Using ROC-curves, the AUC-values for FC counting of all bacteria, only viable bacteria and Gram staining were respectively 0.935, 0.955, and 0.968 (P > 0.05). FC counting of only viable bacteria can predict quickly and reliably positive and negative urine cultures in febrile patients admitted to the emergency department. It can help to improve the speed and accuracy of the diagnostic procedure at the emergency department. © 2017 Clinical Cytometry Society. © 2017 International Clinical Cytometry Society.

  9. Flow cytometric analysis of circulating platelet-monocyte aggregates in whole blood: methodological considerations.

    Science.gov (United States)

    Harding, Scott A; Din, Jehangir N; Sarma, Jaydeep; Jessop, Alasdair; Weatherall, Mark; Fox, Keith A A; Newby, David E

    2007-08-01

    Platelet-monocyte aggregates are increasingly being used to quantify platelet activation. The variables that influence platelet-monocyte aggregates have not been well defined. We sought to determine the effect of blood collection, handling and processing techniques on detected levels of platelet-monocyte aggregates using a flow cytometric assay. Whole blood was labelled with anti-CD14-PE and anti-CD42a-FITC. Thereafter, samples were fixed and red cells lysed. Analysis was performed with the flow cytometer initially triggering on light scatter and then on FL-2 to identify CD14-PE positive monocytes. Platelet-monocyte aggregates were defined as monocytes positive for CD42a. The effect of collection, handling and processing techniques on this assay were assessed. Anticoagulation with heparin (20.1 +/- 2.0%), PPACK (16.8 +/- 1.9%), sodium citrate (12.3 +/- 1.6%) and EDTA (9.5 +/- 1.0%) resulted in markedly different levels of platelet-monocyte aggregation (P venepuncture (20.9 +/- 3.9% vs.13.8 +/- 2.4%, P = 0.03). For every 10 minutes of delay prior to processing platelet-monocyte aggregates increased by 2.8% (P = 0.0001) in PPACK anticoagulated blood and 1.7% (P = 0.01) in citrate anticoagulated blood. Erythrocyte lysis together with fixation does not affect platelet-monocyte aggregation. Platelet-monocyte aggregates remained stable over 24 hours when fixed and stored at 4 degrees C. Multiple handling and processing factors may affect platelet-monocyte aggregation. We recommend the measurement of platelet-monocyte aggregates on samples collected by direct venepuncture, using a direct thrombin inhibitor as the anticoagulant and minimising the time delay before sample fixation.

  10. Flow cytometric immunobead assay for quantitative detection of platelet autoantibodies in immune thrombocytopenia patients.

    Science.gov (United States)

    Zhai, Juping; Ding, Mengyuan; Yang, Tianjie; Zuo, Bin; Weng, Zhen; Zhao, Yunxiao; He, Jun; Wu, Qingyu; Ruan, Changgeng; He, Yang

    2017-10-23

    Platelet autoantibody detection is critical for immune thrombocytopenia (ITP) diagnosis and prognosis. Therefore, we aimed to establish a quantitative flow cytometric immunobead assay (FCIA) for ITP platelet autoantibodies evaluation. Capture microbeads coupled with anti-GPIX, -GPIb, -GPIIb, -GPIIIa and P-selectin antibodies were used to bind the platelet-bound autoantibodies complex generated from plasma samples of 250 ITP patients, 163 non-ITP patients and 243 healthy controls, a fluorescein isothiocyanate (FITC)-conjugated secondary antibody was the detector reagent and mean fluorescence intensity (MFI) signals were recorded by flow cytometry. Intra- and inter-assay variations of the quantitative FCIA assay were assessed. Comparisons of the specificity, sensitivity and accuracy between quantitative and qualitative FCIA or monoclonal antibody immobilization of platelet antigen (MAIPA) assay were performed. Finally, treatment process was monitored by our quantitative FCIA in 8 newly diagnosed ITPs. The coefficient of variations (CV) of the quantitative FCIA assay were respectively 9.4, 3.8, 5.4, 5.1 and 5.8% for anti-GPIX, -GPIb, -GPIIIa, -GPIIb and -P-selectin autoantibodies. Elevated levels of autoantibodies against platelet glycoproteins GPIX, GPIb, GPIIIa, GPIIb and P-selectin were detected by our quantitative FCIA in ITP patients compared to non-ITP patients or healthy controls. The sensitivity, specificity and accuracy of our quantitative assay were respectively 73.13, 81.98 and 78.65% when combining all 5 autoantibodies, while the sensitivity, specificity and accuracy of MAIPA assay were respectively 41.46, 90.41 and 72.81%. A quantitative FCIA assay was established. Reduced levels of platelet autoantibodies could be confirmed by our quantitative FCIA in ITP patients after corticosteroid treatment. Our quantitative assay is not only good for ITP diagnosis but also for ITP treatment monitoring.

  11. FlowFP: A Bioconductor Package for Fingerprinting Flow Cytometric Data

    Directory of Open Access Journals (Sweden)

    Wade T. Rogers

    2009-01-01

    Full Text Available A new software package called flowFP for the analysis of flow cytometry data is introduced. The package, which is tightly integrated with other Bioconductor software for analysis of flow cytometry, provides tools to transform raw flow cytometry data into a form suitable for direct input into conventional statistical analysis and empirical modeling software tools. The approach of flowFP is to generate a description of the multivariate probability distribution function of flow cytometry data in the form of a “fingerprint.” As such, it is independent of a presumptive functional form for the distribution, in contrast with model-based methods such as Gaussian Mixture Modeling. FlowFP is computationally efficient and able to handle extremely large flow cytometry data sets of arbitrary dimensionality. Algorithms and software implementation of the package are described. Use of the software is exemplified with applications to data quality control and to the automated classification of Acute Myeloid Leukemia.

  12. A novel flow cytometric HTS assay reveals functional modulators of ATP binding cassette transporter ABCB6.

    Directory of Open Access Journals (Sweden)

    Kishore Polireddy

    Full Text Available ABCB6 is a member of the adenosine triphosphate (ATP-binding cassette family of transporter proteins that is increasingly recognized as a relevant physiological and therapeutic target. Evaluation of modulators of ABCB6 activity would pave the way toward a more complete understanding of the significance of this transport process in tumor cell growth, proliferation and therapy-related drug resistance. In addition, this effort would improve our understanding of the function of ABCB6 in normal physiology with respect to heme biosynthesis, and cellular adaptation to metabolic demand and stress responses. To search for modulators of ABCB6, we developed a novel cell-based approach that, in combination with flow cytometric high-throughput screening (HTS, can be used to identify functional modulators of ABCB6. Accumulation of protoporphyrin, a fluorescent molecule, in wild-type ABCB6 expressing K562 cells, forms the basis of the HTS assay. Screening the Prestwick Chemical Library employing the HTS assay identified four compounds, benzethonium chloride, verteporfin, tomatine hydrochloride and piperlongumine, that reduced ABCB6 mediated cellular porphyrin levels. Validation of the identified compounds employing the hemin-agarose affinity chromatography and mitochondrial transport assays demonstrated that three out of the four compounds were capable of inhibiting ABCB6 mediated hemin transport into isolated mitochondria. However, only verteporfin and tomatine hydrochloride inhibited ABCB6's ability to compete with hemin as an ABCB6 substrate. This assay is therefore sensitive, robust, and suitable for automation in a high-throughput environment as demonstrated by our identification of selective functional modulators of ABCB6. Application of this assay to other libraries of synthetic compounds and natural products is expected to identify novel modulators of ABCB6 activity.

  13. Air sampling to assess potential generation of aerosolized viable bacteria during flow cytometric analysis of unfixed bacterial suspensions

    Science.gov (United States)

    Carson, Christine F; Inglis, Timothy JJ

    2018-01-01

    This study investigated aerosolized viable bacteria in a university research laboratory during operation of an acoustic-assisted flow cytometer for antimicrobial susceptibility testing by sampling room air before, during and after flow cytometer use. The aim was to assess the risk associated with use of an acoustic-assisted flow cytometer analyzing unfixed bacterial suspensions. Air sampling in a nearby clinical laboratory was conducted during the same period to provide context for the existing background of microorganisms that would be detected in the air. The three species of bacteria undergoing analysis by flow cytometer in the research laboratory were Klebsiella pneumoniae, Burkholderia thailandensis and Streptococcus pneumoniae. None of these was detected from multiple 1000 L air samples acquired in the research laboratory environment. The main cultured bacteria in both locations were skin commensal and environmental bacteria, presumed to have been disturbed or dispersed in laboratory air by personnel movements during routine laboratory activities. The concentrations of bacteria detected in research laboratory air samples were reduced after interventional cleaning measures were introduced and were lower than those in the diagnostic clinical microbiology laboratory. We conclude that our flow cytometric analyses of unfixed suspensions of K. pneumoniae, B. thailandensis and S. pneumoniae do not pose a risk to cytometer operators or other personnel in the laboratory but caution against extrapolation of our results to other bacteria and/or different flow cytometric experimental procedures. PMID:29608197

  14. Fluorescent particles in the antibody solution result in false TF- and CD14-positive microparticles in flow cytometric analysis.

    Science.gov (United States)

    Aass, Hans Christian D; Øvstebø, Reidun; Trøseid, Anne-Marie S; Kierulf, Peter; Berg, Jens Petter; Henriksson, Carola Elisabeth

    2011-12-01

    Tissue factor (TF)-positive microparticles (MPs) are highly procoagulant, and linked to thrombosis in sepsis and cancer. MP-associated TF may be assayed by immunological or functional methods. Several reports have demonstrated discrepancies between TF-protein and TF-activity, which have been explained by antibody binding to "encrypted" or degraded forms of inactive TF-protein. Our goal was to evaluate the possible interference of fluorescent antibody aggregates in solutions containing antibodies against TF and CD14 in flow cytometric analysis. Using monocyte-derived microparticles (MPs) released from human monocytes, incubated with or without lipopolysaccharides (LPS) in vitro, we measured MP-associated TF-protein (flow cytometry) and TF-activity (clot formation assay). MPs released from monocytes exposed to LPS (1 ng mL(-1) ) had ∼14 times higher TF-activity than MPs originated from monocytes exposed to only culture medium. However, using untreated anti-TF antibodies (American Diagnostica and BD) in the flow cytometric analysis, MPs released from unstimulated monocytes had a similar number of TF-positive events as MPs secernated from LPS-stimulated monocytes [∼45,000 events mL(-1) (American Diagnostica); ∼15,000 events mL(-1) (BD)]. These TF-positive events did not exert any TF-activity, and centrifugation (17,000g, 30 min, 4°C) of the antibody solutions prior to use effectively removed the interfering fluorescent events. Removal of fluorescent interference, probably in the form of fluorescent antibody aggregates, from the antibody solutions by centrifugation is essential to prevent the occurrence of false positive flow cytometric events. The events can be mistaken as MP-associated TF-protein, and interpreted as a discrepancy between TF-protein and TF-activity. Copyright © 2011 International Society for Advancement of Cytometry.

  15. A short-term in vitro test for tumour sensitivity to adriamycin based on flow cytometric DNA analysis

    DEFF Research Database (Denmark)

    Engelholm, S A; Spang-Thomsen, M; Vindeløv, L L

    1983-01-01

    A new method to test the sensitivity of tumour cells to chemotherapy is presented. Tumour cells were incubated in vitro on agar, and drug-induced cell cycle perturbation was monitored by flow cytometric DNA analysis. In the present study the method was applied to monitor the effect of adriamycin...... tumours. The dose level causing maximum accumulation in the G2 + M phase is suggested as a parameter for quantifying the sensitivity. The results indicate that the method can be extended to sensitivity testing of human tumours....

  16. Development of a flow cytometric assay to quantify lymphocyte adhesion to cytokine-stimulated human endothelial and biliary epithelial cells.

    Science.gov (United States)

    Korlipara, L V; Leon, M P; Rix, D A; Douglas, M S; Gibbs, P; Bassendine, M F; Kirby, J A

    1996-05-27

    The adhesive interaction between T lymphocytes and parenchymal cells is of importance for many processes of the cellular immune response. This adhesion is regulated by the activation status of the T cell and by cytokines in the microenvironment which can alter adhesion molecule expression by endothelial and epithelial cells. In this study results from an isotopic adhesion assay were compared with those from a flow cytometric assay in order to determine which was most appropriate for the investigation of lymphocyte adhesion to human umbilical vein endothelial cells (HUVEC) and intrahepatic biliary epithelial cells (HIBEC). Treatment of both these cell types with the proinflammatory cytokines interferon-gamma (IFN-gamma) or tumour necrosis factor-alpha (TNF-alpha) significantly upregulated expression of intercellular adhesion molecule-1 (ICAM-1). Treatment with TNF-alpha also induced endothelial cells to express vascular cell adhesion molecule-1 (VCAM-1). The isotopic assay demonstrated increased adhesion of lymphoblasts to HUVEC which had been stimulated with cytokines for 15 h but failed to detect major changes in adhesion following 72 h of cytokine treatment of HUVEC or HIBEC. However, the flow cytometric assay reproducibly demonstrated increased adhesion following cytokine treatment for both these time periods; these increases corresponded with the changes in adhesion molecule expression by cytokine-stimulated HUVEC and HIBEC targets. The differences in apparent adhesion measured by the two assays after cytokine stimulation for 72 h may be explained by cytokine-induced changes in the morphology and confluency of cultured cells. Results of the isotopic assay are proportional to the total number of lymphoid cells bound by the cultured target cells and will be distorted by changes in effective target cell area. The flow cytometric assay measures the mean number of lymphoid cells bound by each target cell and is independent of the total binding area. It is concluded

  17. Flow cytometric analysis of expression of interleukin-2 receptor beta chain (p70-75) on various leukemic cells

    International Nuclear Information System (INIS)

    Hoshino, S.; Oshimi, K.; Tsudo, M.; Miyasaka, M.; Teramura, M.; Masuda, M.; Motoji, T.; Mizoguchi, H.

    1990-01-01

    We analyzed the expression of the interleukin-2 receptor (IL-2R) beta chain (p70-75) on various leukemic cells from 44 patients by flow cytometric analysis using the IL-2R beta chain-specific monoclonal antibody, designated Mik-beta 1. Flow cytometric analysis demonstrated the expression of the IL-2R beta chain on granular lymphocytes (GLs) from all eight patients with granular lymphocyte proliferative disorders (GLPDs), on adult T-cell leukemia (ATL) cells from all three patients with ATL, and on T-cell acute lymphoblastic leukemia (T-ALL) cells from one of three patients with T-ALL. Although GLs from all the GLPD patients expressed the IL-2R beta chain alone and not the IL-2R alpha chain (Tac-antigen: p55), ATL and T-ALL cells expressing the beta chain coexpressed the alpha chain. In two of seven patients with common ALL (cALL) and in both patients with B-cell chronic lymphocytic leukemia, the leukemic cells expressed the alpha chain alone. Neither the alpha chain nor the beta chain was expressed on leukemic cells from the remaining 28 patients, including all 18 patients with acute nonlymphocytic leukemia, five of seven patients with cALL, all three patients with multiple myeloma, and two of three patients with T-ALL. These results indicate that three different forms of IL-2R chain expression exist on leukemic cells: the alpha chain alone; the beta chain alone; and both the alpha and beta chains. To examine whether the results obtained by flow cytometric analysis actually reflect functional aspects of the expressed IL-2Rs, we studied the specific binding of 125I-labeled IL-2 (125I-IL-2) to leukemic cells in 18 of the 44 patients. In addition, we performed 125I-IL-2 crosslinking studies in seven patients. The results of IL-2R expression of both 125I-IL-2 binding assay and crosslinking studies were in agreement with those obtained by flow cytometric analysis

  18. Micronuclei frequency in circulating erythrocytes from rainbow trout (Oncorhynchus mykiss) subjected to radiation, an image analysis and flow cytometric study

    International Nuclear Information System (INIS)

    Schultz, N.; Norrgren, L.; Grawe, J.; Johannisson, A.; Medhage, O.

    1993-01-01

    Rainbow trout (oncorhynchus mykiss) were exposed to a single X-ray dose of 4 Gy. The frequency of micronuclei in the peripheral erythrocytes was investigated at regular intervals up to 58 days after the exposure. A flow cytometric method and a semi-automatic image analysis method were used to estimate the micronuclei frequency. The results show that both methods can detect an increased frequency of micronuclei in peripheral erythrocytes from exposed fish. However, the semi-automatic image analysis method was the most stable and sensitive. (Author)

  19. Image cytometric evaluation of nuclear texture features and DNA content of the reticular form of oral lichen planus.

    Science.gov (United States)

    Rode, Matjaz; Flezar, Margareta Strojan; Kogoj-Rode, Mirela; Us-Krasovec, Marija

    2006-10-01

    To analyze image cytometric chromatin changes reflected in nuclear texture features and DNA ploidy of oral lichen planus in relation to the normal buccal mucosa and buccal mucosa expressing malignancy-associated changes in cancer patients. Twenty-eight patients with the reticular form of oral lichen planus, with a follow-up period of 25 years, 50 healthy controls and 50 lung cancer patients were included in the study. Scrapings of buccal mucosa were suspended in transport medium. Monolayer filter preparations were Feulgen-thionin stained. Image cytometric analysis was performed by Cyto-Savant. All oral lichen planus specimens in our study were diploid. In univariate analysis, differences between the normal buccal mucosa and oral lichen planus were found in several nuclear texture features, which gave an 80% correct classification rate in multivariate analysis. In the second part of the study, the classifier that recognizes malignancy-associated changes on the buccal mucosa of patients with lung cancer correctly recognized > 80% of oral lichen planus samples as normal buccal mucosa. Our results indicate that chromatin changes in oral lichen planus exist compared to normal cells; however, the chromatin structure of the reticular form of oral lichen planus does not express malignancy-associated changes and is more similar to normal squamous cells.

  20. Clonal evolution demonstrated by flow cytometric DNA analysis of a human colonic carcinoma grown in nude mice

    DEFF Research Database (Denmark)

    Vindeløv, L L; Spang-Thomsen, M; Visfeldt, J

    1982-01-01

    A spontaneous change in DNA content of a human colonic carcinoma grown in nude mice was observed fortuitously. The tumor initially had a G1 cell DNA content of 1.3 times that of normal cells. Flow cytometric DNA analysis showed in transplant generation 56 the appearance of a new subpopulation whi...... evolution of a tumor would be less pronounced if old subpopulations often become extinct as new ones emerge. Heterogeneity of human tumors is of clinical importance because the individual subpopulations may have different sensitivity patterns to antineoplastic drugs.......A spontaneous change in DNA content of a human colonic carcinoma grown in nude mice was observed fortuitously. The tumor initially had a G1 cell DNA content of 1.3 times that of normal cells. Flow cytometric DNA analysis showed in transplant generation 56 the appearance of a new subpopulation which...... in three passages completely overgrew the original population. The DNA content of the new subpopulation was twice that of the original population. The observation supports the hypothesis of clonal evolution of tumor cell populations. The growth rates of the tumor before and after the change showed...

  1. Rapid Detection and Enumeration of Giardia lamblia Cysts in Water Samples by Immunomagnetic Separation and Flow Cytometric Analysis ▿ †

    Science.gov (United States)

    Keserue, Hans-Anton; Füchslin, Hans Peter; Egli, Thomas

    2011-01-01

    Giardia lamblia is an important waterborne pathogen and is among the most common intestinal parasites of humans worldwide. Its fecal-oral transmission leads to the presence of cysts of this pathogen in the environment, and so far, quantitative rapid screening methods are not available for various matrices, such as surface waters, wastewater, or food. Thus, it is necessary to establish methods that enable reliable rapid detection of a single cyst in 10 to 100 liters of drinking water. Conventional detection relies on cyst concentration, isolation, and confirmation by immunofluorescence microscopy (IFM), resulting in low recoveries and high detection limits. Many different immunomagnetic separation (IMS) procedures have been developed for separation and cyst purification, so far with variable but high losses of cysts. A method was developed that requires less than 100 min and consists of filtration, resuspension, IMS, and flow cytometric (FCM) detection. MACS MicroBeads were used for IMS, and a reliable flow cytometric detection approach was established employing 3 different parameters for discrimination from background signals, i.e., green and red fluorescence (resulting from the distinct pattern emitted by the fluorescein dye) and sideward scatter for size discrimination. With spiked samples, recoveries exceeding 90% were obtained, and false-positive results were never encountered for negative samples. Additionally, the method was applicable to naturally occurring cysts in wastewater and has the potential to be automated. PMID:21685159

  2. Flow cytometric measurement of RNA synthesis using bromouridine labelling and bromodeoxyuridine antibodies

    DEFF Research Database (Denmark)

    Jensen, P O; Larsen, J; Christiansen, J

    1993-01-01

    Nuclear RNA synthesis can be analysed by flow cytometry of cells labelled with 5-bromouridine (BrUrd) and stained with anti-bromodeoxyuridine (BrdUrd) antibody and FITC-conjugated secondary antibody. A panel of 5 different commercially available anti-BrdUrd antibodies was tested on cells of a HL-60...... human leukemia cell line, stained as a methanol-fixed nuclear suspension. The BrUrd-induced fluorescence signals were highest with the antibody ABDM (Partec), moderate but reproducible with B-44 (Becton Dickinson), variable or low with BR-3 and IU-4 (Caltag), and not detectable with Bu20a (DAKO...... the variation of RNA synthesis during the cell cycle. The BrUrd incorporation was high in the S and G2 phase, variable in G1, and negligible in mitosis. Similar results were obtained using other cell types....

  3. A Protocol for the Comprehensive Flow Cytometric Analysis of Immune Cells in Normal and Inflamed Murine Non-Lymphoid Tissues

    Science.gov (United States)

    Yu, Yen-Rei A.; O’Koren, Emily G.; Hotten, Danielle F.; Kan, Matthew J.; Kopin, David; Nelson, Erik R.; Que, Loretta; Gunn, Michael D.

    2016-01-01

    Flow cytometry is used extensively to examine immune cells in non-lymphoid tissues. However, a method of flow cytometric analysis that is both comprehensive and widely applicable has not been described. We developed a protocol for the flow cytometric analysis of non-lymphoid tissues, including methods of tissue preparation, a 10-fluorochrome panel for cell staining, and a standardized gating strategy, that allows the simultaneous identification and quantification of all major immune cell types in a variety of normal and inflamed non-lymphoid tissues. We demonstrate that our basic protocol minimizes cell loss, reliably distinguishes macrophages from dendritic cells (DC), and identifies all major granulocytic and mononuclear phagocytic cell types. This protocol is able to accurately quantify 11 distinct immune cell types, including T cells, B cells, NK cells, neutrophils, eosinophils, inflammatory monocytes, resident monocytes, alveolar macrophages, resident/interstitial macrophages, CD11b- DC, and CD11b+ DC, in normal lung, heart, liver, kidney, intestine, skin, eyes, and mammary gland. We also characterized the expression patterns of several commonly used myeloid and macrophage markers. This basic protocol can be expanded to identify additional cell types such as mast cells, basophils, and plasmacytoid DC, or perform detailed phenotyping of specific cell types. In examining models of primary and metastatic mammary tumors, this protocol allowed the identification of several distinct tumor associated macrophage phenotypes, the appearance of which was highly specific to individual tumor cell lines. This protocol provides a valuable tool to examine immune cell repertoires and follow immune responses in a wide variety of tissues and experimental conditions. PMID:26938654

  4. A flow-cytometric NK-cytotoxicity assay adapted for use in rat repeated dose toxicity studies

    International Nuclear Information System (INIS)

    Marcusson-Staahl, Maritha; Cederbrant, Karin

    2003-01-01

    A recent regulatory document for immunotoxicity testing of new pharmaceutical drugs includes cytotoxic natural killer (NK)-cell function as a required parameter in repeated dose toxicity studies. The classical 51 Cr-release assay is the conventional test for cytotoxicity testing but several drawbacks with this assay has increased the demand for new reliable test systems. Here, we describe the optimisation of a flow-cytometric cytotoxicity assay especially adapted for regulatory rat studies in drug development. The test principle is based on target cell labelling with 5-(6)-carboxy-fluorescein succinimidyl ester (CFSE) and subsequent DNA-labelling with propidium iodide (PI) for identification of target cells with compromised cell membranes. The results are expressed as percentage of dead targets on a cell-to-cell basis. The final format of the assay includes 0.5 ml peripheral blood, 1.25x10 5 effector cells per sample, and collection of 500 target events by flow-cytometry. When NKR-P1+ cells were removed from the effector cell population by magnetic depletion the relative proportion decreased from 6 to 0.08%. The corresponding cytotoxic activity decreased from 68 to 8%. Also, the cytotoxic activity showed a significant and positive correlation with the proportion of NK-cells present in the effector cell suspension. Thus, the cytotoxicity measured is almost exclusively exerted by NK-cells. The current flow-cytometric test benefits from using peripheral blood as a source for effector cells since it will not conflict with the use of spleen for histopathological investigations in repeated dose toxicity studies. Additionally, since only a minimal number of effector cells are required per sample repeated testing of the same animal is enabled

  5. A Protocol for the Comprehensive Flow Cytometric Analysis of Immune Cells in Normal and Inflamed Murine Non-Lymphoid Tissues.

    Directory of Open Access Journals (Sweden)

    Yen-Rei A Yu

    Full Text Available Flow cytometry is used extensively to examine immune cells in non-lymphoid tissues. However, a method of flow cytometric analysis that is both comprehensive and widely applicable has not been described. We developed a protocol for the flow cytometric analysis of non-lymphoid tissues, including methods of tissue preparation, a 10-fluorochrome panel for cell staining, and a standardized gating strategy, that allows the simultaneous identification and quantification of all major immune cell types in a variety of normal and inflamed non-lymphoid tissues. We demonstrate that our basic protocol minimizes cell loss, reliably distinguishes macrophages from dendritic cells (DC, and identifies all major granulocytic and mononuclear phagocytic cell types. This protocol is able to accurately quantify 11 distinct immune cell types, including T cells, B cells, NK cells, neutrophils, eosinophils, inflammatory monocytes, resident monocytes, alveolar macrophages, resident/interstitial macrophages, CD11b- DC, and CD11b+ DC, in normal lung, heart, liver, kidney, intestine, skin, eyes, and mammary gland. We also characterized the expression patterns of several commonly used myeloid and macrophage markers. This basic protocol can be expanded to identify additional cell types such as mast cells, basophils, and plasmacytoid DC, or perform detailed phenotyping of specific cell types. In examining models of primary and metastatic mammary tumors, this protocol allowed the identification of several distinct tumor associated macrophage phenotypes, the appearance of which was highly specific to individual tumor cell lines. This protocol provides a valuable tool to examine immune cell repertoires and follow immune responses in a wide variety of tissues and experimental conditions.

  6. Flow-cytometric analysis of changes in lymphocyte subsets in the blood of cancer patients during radiation therapy

    International Nuclear Information System (INIS)

    Ogawa, Yasuhiro; Maeda, Tomoho; Ogawa, Yukiko

    1983-01-01

    In this study, we report a clinical application of a rapid flow-cytometric immunofluorescence method for studying lymphocyte subsets in whole blood by using monoclonal antibodies OKT3, OKT4, OKT8, OKIa1, OKT6, OKT10, OKT9, OKT11 and OKM1. Changes in the relative percentages and absolute counts of various lymphocyte subsets during radiotherapy of cancer patients were evaluated by this method. In patients with good response, the elevation of OKT 4 + -lymphocyte level and/or the depression of OKT8 + -lymphocyte level was observed. In contrast, the depression of OKT4 + -lymphocyte level and/or the elevation of OKT8 + -lymphocyte level was observed on patients with poor response. Our data indicate that these monoclonal antibodies can be regarded as invaluable tools for evaluation of immune status of cancer patients during radiotherapy. (author)

  7. Flow cytometric evaluation of physico-chemical impact on Gram-positive and Gram-negative bacteria

    Directory of Open Access Journals (Sweden)

    Antje eFröhling

    2015-09-01

    Full Text Available Since heat sensitivity of fruits and vegetables limits the application of thermal inactivation processes, new emerging inactivation technologies have to be established to fulfil the requirements of food safety without affecting the produce quality. The efficiency of inactivation treatments has to be ensured and monitored. Monitoring of inactivation effects is commonly performed using traditional cultivation methods which have the disadvantage of the time span needed to obtain results.The aim of this study was to compare the inactivation effects of peracetic acid (PAA, ozonated water (O3 and cold atmospheric pressure plasma (CAPP on Gram-positive and Gram-negative bacteria using flow cytometric methods. E. coli cells were completely depolarized after treatment (15 s with 0.25 % PAA at 10 °C, and after treatment (10 s with 3.8 mg l-1 O3 at 12°C. The membrane potential of CAPP treated cells remained almost constant at an operating power of 20 W over a time period of 3 min, and subsequently decreased within 30 s of further treatment. Complete membrane permeabilization was observed after 10 s O3 treatment, but treatment with PAA and CAPP did not completely permeabilize the cells within 2 min and 4 min, respectively. Similar results were obtained for esterase activity. O3 inactivates cellular esterase but esterase activity was detected after 4 min CAPP treatment and 2 min PAA treatment. L. innocua cells and P. carotovorum cells were also permeabilized instantaneously by O3 treatment at concentrations of 3.8 ± 1 mg l-1. However, higher membrane permeabilization of L. innocua and P. carotovorum than of E. coli was observed at CAPP treatment of 20 W. The degree of bacterial damage due to the inactivation processes is highly dependent on treatment parameters as well as on treated bacteria. Important information regarding the inactivation mechanisms can be obtained by flow cytometric measurements and this enables the definition of critical process

  8. Effect of 17 beta-oestradiol on growth curves and flow cytometric DNA distribution of two human breast carcinomas grown in nude mice

    DEFF Research Database (Denmark)

    Brünner, N; Spang-Thomsen, M; Vindeløv, L

    1983-01-01

    The effect of 17 beta-oestradiol on a "receptor positive" and on a "receptor negative" human breast carcinoma grown in nude mice was studied. Experimental growth data were used to determine the effect on tumour growth. Flow cytometric DNA analysis (FCM) performed on tumour tissue obtained...

  9. A heparin-based method for flow cytometric analysis of microparticles directly from platelet-poor plasma in calcium containing buffer

    DEFF Research Database (Denmark)

    Iversen, Line V; Ostergaard, Ole; Nielsen, Christoffer

    2013-01-01

    V negative MPs to clinical parameters in systemic diseases, which emphasize the importance of including characterization of AnxV-binding. An obstacle in flow cytometric analysis of AnxV-binding to MPs is that plasma may clot when adding the Ca(2+)-containing buffers. We here devise a simple method...

  10. Clinical flow cytometric screening of SAP and XIAP expression accurately identifies patients with SH2D1A and XIAP/BIRC4 mutations.

    Science.gov (United States)

    Gifford, Carrie E; Weingartner, Elizabeth; Villanueva, Joyce; Johnson, Judith; Zhang, Kejian; Filipovich, Alexandra H; Bleesing, Jack J; Marsh, Rebecca A

    2014-07-01

    X-linked lymphoproliferative disease is caused by mutations in two genes, SH2D1A and XIAP/BIRC4. Flow cytometric methods have been developed to detect the gene products, SAP and XIAP. However, there is no literature describing the accuracy of flow cytometric screening performed in a clinical lab setting. We reviewed the clinical flow cytometric testing results for 656 SAP and 586 XIAP samples tested during a 3-year period. Genetic testing was clinically performed as directed by the managing physician in 137 SAP (21%) and 115 XIAP (20%) samples. We included these samples for analyses of flow cytometric test accuracy. SH2D1A mutations were detected in 15/137 samples. SAP expression was low in 13/15 (sensitivity 87%, CI 61-97%). Of the 122 samples with normal sequencing, SAP was normal in 109 (specificity 89%, CI 82-94%). The positive predictive values (PPVs) and the negative predictive values (NPVs) were 50% and 98%, respectively. XIAP/BIRC4 mutations were detected in 19/115 samples. XIAP expression was low in 18/19 (sensitivity 95%, CI 73-100%). Of the 96 samples with normal sequencing, 59 had normal XIAP expression (specificity 61%, CI 51-71%). The PPVs and NPVs were 33% and 98%, respectively. Receiver-operating characteristic analysis was able to improve the specificity to 75%. Clinical flow cytometric screening tests for SAP and XIAP deficiencies offer good sensitivity and specificity for detecting genetic mutations, and are characterized by high NPVs. We recommend these tests for patients suspected of having X-linked lymphoproliferative disease type 1 (XLP1) or XLP2. © 2014 Clinical Cytometry Society.

  11. Development of a novel flow cytometric approach to evaluate fish sperm chromatin using fixed samples

    Science.gov (United States)

    Jenkins, Jill A.

    2013-01-01

    The integrity of the paternal DNA is essential for the accurate transmission of genetic information, yet fertilization is not inhibited by chromatin breakage. Some methods are available for the sensitive detection of DNA damage and can be applied in studies of environmental toxicology, carcinogenesis, aging, and assisted reproduction techniques in both clinical and experimental settings. Because semen samples obtained from remote locations undergo chromatin damage prior to laboratory assessment, the present study was undertaken to evaluate treatments for effective chromatin staining in the development of a DNA fragmentation assay using fixed milt from yellow perch (Perca flavescens). Similar to the sperm chromatin structure assay (SCSA), susceptibility of nuclear DNA to acid-induced denaturation was measured by flow cytometry (FCM). Use of 10% buffered formalin for milt fixation allowed easier peak discrimination than 4% paraformaldehyde. The effects of time and temperature of incubation in 0.08 N HCl were evaluated in order to determine the ideal conditions for promoting DNA decondensation and making strand breaks more available for staining and detection by FCM. The best results were obtained with incubation at 37°C for 1 minute, followed by cold propidium iodide staining for 30 minutes.

  12. Flow-cytometric assessment of cellular poly(ADP-ribosylation capacity in peripheral blood lymphocytes

    Directory of Open Access Journals (Sweden)

    Hyland Paul

    2006-07-01

    Full Text Available Abstract Background Poly(ADP-ribosylation is a posttranslational modification of nuclear proteins catalysed by poly(ADP-ribose polymerases (PARPs, using NAD+ as a substrate. Activation of PARP-1 is in immediate response to DNA damage generated by endogenous and exogenous damaging agents. It has been implicated in several crucial cellular processes including DNA repair and maintenance of genomic stability, which are both intimately linked with the ageing process. The measurement of cellular poly(ADP-ribosylation capacity, defined as the amount of poly(ADP-ribose produced under maximal stimulation, is therefore relevant for research on ageing, as well as for a variety of other scientific questions. Results This paper reports a new, robust protocol for the measurement of cellular poly(ADP-ribosylation capacity in PBMC or Jurkat T-cells using flow cytometry, based on a previously established immuno-dot-blot assay. In order to validate the new assay, we determined the dose-response curve of 3-aminobenzamide, a well-known competitive PARP inhibitor, and we derived an IC50 that is very close to the published value. When testing a set of PBMC samples taken from fifteen healthy young human donors, we could confirm the presence of a substantial interindividual variation, as previously observed using a radiometric assay. Conclusion The methodology described in this paper should be generally useful for the determination of cellular poly(ADP-ribosylation capacity in a wide variety of settings, especially for the comparison of large sets of samples, such as population studies. In contrast to previously published radiometric or immuno-dot-blot assays, the new FACS-based method allows (i selective analysis of mononuclear cells by gating and (ii detection of a possible heterogeneity in poly(ADP-ribosylation capacity between cells of the same type.

  13. CD33 monoclonal antibody conjugated Au cluster nano-bioprobe for targeted flow-cytometric detection of acute myeloid leukaemia

    Energy Technology Data Exchange (ETDEWEB)

    Retnakumari, Archana; Jayasimhan, Jasusri; Chandran, Parwathy; Menon, Deepthy; Nair, Shantikumar; Mony, Ullas; Koyakutty, Manzoor, E-mail: manzoork@aims.amrita.edu, E-mail: ullasmony@aims.amrita.edu [Amrita Centre for Nanoscience and Molecular Medicine, Amrita Institute of Medical Science, Cochin 682 041 (India)

    2011-07-15

    Protein stabilized gold nanoclusters (Au-NCs) are biocompatible, near-infrared (NIR) emitting nanosystems having a wide range of biomedical applications. Here, we report the development of a Au-NC based targeted fluorescent nano-bioprobe for the flow-cytometric detection of acute myeloid leukaemia (AML) cells. Au-NCs with {approx} 25-28 atoms showing bright red-NIR fluorescence (600-750 nm) and average size of {approx} 0.8 nm were prepared by bovine serum albumin assisted reduction-cum-stabilization in aqueous phase. The protein protected clusters were conjugated with monoclonal antibody against CD33 myeloid antigen, which is overexpressed in {approx} 99.2% of the primitive population of AML cells, as confirmed by immunophenotyping using flow cytometry. Au-NC-CD33 conjugates having average size of {approx} 12 nm retained bright fluorescence over an extended duration of {approx} a year, as the albumin protein protects Au-NCs against degradation. Nanotoxicity studies revealed excellent biocompatibility of Au-NC conjugates, as they showed no adverse effect on the cell viability and inflammatory response. Target specificity of the conjugates for detecting CD33 expressing AML cells (KG1a) in flow cytometry showed specific staining of {approx} 95.4% of leukaemia cells within 1-2 h compared to a non-specific uptake of {approx} 8.2% in human peripheral blood cells (PBMCs) which are CD33{sup low}. The confocal imaging also demonstrated the targeted uptake of CD33 conjugated Au-NCs by leukaemia cells, thus confirming the flow cytometry results. This study demonstrates that novel nano-bioprobes can be developed using protein protected fluorescent nanoclusters of Au for the molecular receptor targeted flow cytometry based detection and imaging of cancer cells.

  14. CD33 monoclonal antibody conjugated Au cluster nano-bioprobe for targeted flow-cytometric detection of acute myeloid leukaemia

    Science.gov (United States)

    Retnakumari, Archana; Jayasimhan, Jasusri; Chandran, Parwathy; Menon, Deepthy; Nair, Shantikumar; Mony, Ullas; Koyakutty, Manzoor

    2011-07-01

    Protein stabilized gold nanoclusters (Au-NCs) are biocompatible, near-infrared (NIR) emitting nanosystems having a wide range of biomedical applications. Here, we report the development of a Au-NC based targeted fluorescent nano-bioprobe for the flow-cytometric detection of acute myeloid leukaemia (AML) cells. Au-NCs with ~ 25-28 atoms showing bright red-NIR fluorescence (600-750 nm) and average size of ~ 0.8 nm were prepared by bovine serum albumin assisted reduction-cum-stabilization in aqueous phase. The protein protected clusters were conjugated with monoclonal antibody against CD33 myeloid antigen, which is overexpressed in ~ 99.2% of the primitive population of AML cells, as confirmed by immunophenotyping using flow cytometry. Au-NC-CD33 conjugates having average size of ~ 12 nm retained bright fluorescence over an extended duration of ~ a year, as the albumin protein protects Au-NCs against degradation. Nanotoxicity studies revealed excellent biocompatibility of Au-NC conjugates, as they showed no adverse effect on the cell viability and inflammatory response. Target specificity of the conjugates for detecting CD33 expressing AML cells (KG1a) in flow cytometry showed specific staining of ~ 95.4% of leukaemia cells within 1-2 h compared to a non-specific uptake of ~ 8.2% in human peripheral blood cells (PBMCs) which are CD33low. The confocal imaging also demonstrated the targeted uptake of CD33 conjugated Au-NCs by leukaemia cells, thus confirming the flow cytometry results. This study demonstrates that novel nano-bioprobes can be developed using protein protected fluorescent nanoclusters of Au for the molecular receptor targeted flow cytometry based detection and imaging of cancer cells.

  15. CD33 monoclonal antibody conjugated Au cluster nano-bioprobe for targeted flow-cytometric detection of acute myeloid leukaemia

    International Nuclear Information System (INIS)

    Retnakumari, Archana; Jayasimhan, Jasusri; Chandran, Parwathy; Menon, Deepthy; Nair, Shantikumar; Mony, Ullas; Koyakutty, Manzoor

    2011-01-01

    Protein stabilized gold nanoclusters (Au-NCs) are biocompatible, near-infrared (NIR) emitting nanosystems having a wide range of biomedical applications. Here, we report the development of a Au-NC based targeted fluorescent nano-bioprobe for the flow-cytometric detection of acute myeloid leukaemia (AML) cells. Au-NCs with ∼ 25-28 atoms showing bright red-NIR fluorescence (600-750 nm) and average size of ∼ 0.8 nm were prepared by bovine serum albumin assisted reduction-cum-stabilization in aqueous phase. The protein protected clusters were conjugated with monoclonal antibody against CD33 myeloid antigen, which is overexpressed in ∼ 99.2% of the primitive population of AML cells, as confirmed by immunophenotyping using flow cytometry. Au-NC-CD33 conjugates having average size of ∼ 12 nm retained bright fluorescence over an extended duration of ∼ a year, as the albumin protein protects Au-NCs against degradation. Nanotoxicity studies revealed excellent biocompatibility of Au-NC conjugates, as they showed no adverse effect on the cell viability and inflammatory response. Target specificity of the conjugates for detecting CD33 expressing AML cells (KG1a) in flow cytometry showed specific staining of ∼ 95.4% of leukaemia cells within 1-2 h compared to a non-specific uptake of ∼ 8.2% in human peripheral blood cells (PBMCs) which are CD33 low . The confocal imaging also demonstrated the targeted uptake of CD33 conjugated Au-NCs by leukaemia cells, thus confirming the flow cytometry results. This study demonstrates that novel nano-bioprobes can be developed using protein protected fluorescent nanoclusters of Au for the molecular receptor targeted flow cytometry based detection and imaging of cancer cells.

  16. Immuno-flow cytometric detection of the ichthyotoxic dinoflagellates Gyrodinium aureolum and Gymnodinium nagasakiense: independence of physiological state

    Science.gov (United States)

    Vrieling, Engel G.; van de Poll, Willem H.; Vriezekolk, Gertie; Gieskes, Winfried W. C.

    1997-05-01

    The ichthyotoxic dinoflagellates Gyrodinium aureolum and Gymnodinium nagasakiense were cultured under different environmental conditions to test possible variability in immunochemical labelling intensity of cell-surface antigens using species-specific monoclonal antibodies. Variation of antigen abundance (which is directly related to labelling intensity) at the cell surface, determined by immuno-flow cytometry of cells labelled with FITC, appeared to be small but significant compared to control cultures. In general, a minor decrease in FIX fluorescence was recorded during exponential growth, followed by an increase during stationary growth. FITC fluorescence was correlated with cell size, shape and structure. This suggests a constant number of antigens per unit of cell surface. In all cultures, immunochemically labelled cells were distinguished clearly from unlabelled cells; immuno-flow cytometric identification is apparently not affected by growth conditions. Only at the end of the stationary growth phase in batch cultures did the FITC fluorescence values drop, which suggests that unhealthy, dying or lysing cells may either alter the composition of the cell surface or just fail to express the antigen.

  17. An improved flow cytometric immunobead array to detect autoantibodies in plasma from patients with immune thrombocytopenic purpura.

    Science.gov (United States)

    Zhao, Yunxiao; Zhu, Mingqing; Jiang, Miao; Zuo, Bin; Wu, Qingyu; Ruan, Changgeng; He, Yang

    2015-01-01

    Autoantibodies against platelet glycoproteins (GPs) play an important role in immune thrombocytopenic purpura (ITP). This study was to develop an improved flow cytometric immunobead array (FCIA) assay to detect platelet autoantibodies in ITP patient plasma. Plasma samples were isolated from 71 ITP patients and 136 non-ITP controls and incubated with platelets from normal individuals. After washing, platelets were lysed and the platelet lysates were incubated with polystyrene microbeads coupled with monoclonal antibodies against human GPs IX (SZ1), Ib (SZ2), IIIa (SZ21), IIb (SZ22), and P-selectin (SZ51). Platelet GP-autoantibody complexes were detected by flow cytometry using a FITC-labeled secondary antibody. Autoantibodies against platelet GPIb, GPIIb, GPIIIa, GPIX and P-selectin were detected in plasma from ITP patients, as indicated by high mean fluorescent intensity values when microbeads with antibodies SZ1, SZ2, SZ21, SZ22, and SZ51 were used. In ROC analysis, values of the area under the curve were 0.74, 0.83, 0.80, 0.79 and 0.87, respectively. Compared with the previously reported assays, this new FCIA eliminated the need of isolating platelets from ITP patients without compromising assay sensitivity and accuracy in predicting ITP. This simplified FICA assay may be more suitable for ITP diagnosis in clinical laboratory settings. Copyright © 2014 Elsevier B.V. All rights reserved.

  18. Flow cytometric-membrane potential detection of sodium channel active marine toxins: application to ciguatoxins in fish muscle and feasibility of automating saxitoxin detection.

    Science.gov (United States)

    Manger, Ronald; Woodle, Doug; Berger, Andrew; Dickey, Robert W; Jester, Edward; Yasumoto, Takeshi; Lewis, Richard; Hawryluk, Timothy; Hungerford, James

    2014-01-01

    Ciguatoxins are potent neurotoxins with a significant public health impact. Cytotoxicity assays have allowed the most sensitive means of detection of ciguatoxin-like activity without reliance on mouse bioassays and have been invaluable in studying outbreaks. An improvement of these cell-based assays is presented here in which rapid flow cytometric detection of ciguatoxins and saxitoxins is demonstrated using fluorescent voltage sensitive dyes. A depolarization response can be detected directly due to ciguatoxin alone; however, an approximate 1000-fold increase in sensitivity is observed in the presence of veratridine. These results demonstrate that flow cytometric assessment of ciguatoxins is possible at levels approaching the trace detection limits of our earlier cytotoxicity assays, however, with a significant reduction in analysis time. Preliminary results are also presented for detection of brevetoxins and for automation and throughput improvements to a previously described method for detecting saxitoxins in shellfish extracts.

  19. A flow cytometric method for characterization of circulating cell-derived microparticles in plasma

    DEFF Research Database (Denmark)

    Nielsen, Morten Hjuler; Beck-Nielsen, Henning; Andersen, Morten Nørgaard

    2014-01-01

    BACKGROUND AND AIM: Previous studies on circulating microparticles (MPs) indicate that the majority of MPs are of a size below the detection limit of most standard flow cytometers. The objective of the present study was to establish a method to analyze MP subpopulations above the threshold...... of detection of a new generation BD FACSAria™ III digital flow cytometer. METHODS: We analyzed MP subpopulations in plasma from 24 healthy individuals (9 males and 15 females). MPs were identified according to their size (

  20. Simple and easy method to evaluate uptake potential of nanoparticles in mammalian cells using a flow cytometric light scatter analysis.

    Science.gov (United States)

    Suzuki, Hiroshi; Toyooka, Tatsushi; Ibuki, Yuko

    2007-04-15

    Many classes of nanoparticles have been synthesized and widely applied, however, there is a serious lack of information concerning their effects on human health and the environment. Considering that their use will increase, accurate and cost-effective measurement techniques for characterizing "nanotoxicity" are required. One major toxicological concern is that nanoparticles are easily taken up in the human body. In this study, we developed a method of evaluating the uptake potential of nanosized particles using flow cytometric light scatter. Suspended titanium dioxide (TiO2) particles (5, 23, or 5000 nm) were added to Chinese hamster ovary cells. Observation by confocal laser scanning microscopy showed that the TiO2 particles easily moved to the cytoplasm of the cultured mammalian cells, not to the nucleus. The intensity of the side-scattered light revealed that the particles were taken up in the cells dose-, time-, and size-dependently. In addition, surface-coating of TiO2 particles changed the uptake into the cells, which was accurately reflected in the intensity of the side-scattered light. The uptake of other nanoparticles such as silver (Ag) and iron oxide (Fe3O4) also could be detected. This method could be used for the initial screening of the uptake potential of nanoparticles as an index of "nanotoxicity".

  1. Flow Cytometric Evaluation of Human Neutrophil Apoptosis During Nitric Oxide Generation In Vitro: The Role of Exogenous Antioxidants

    Directory of Open Access Journals (Sweden)

    Zofia Sulowska

    2005-01-01

    in vitro. The effect of exogenous supply of NO donors such as SNP, SIN-1, and GEA-3162 on the course of human neutrophil apoptosis and the role of extracellular antioxidants in this process was investigated. Isolated from peripheral blood, neutrophils were cultured in the presence or absence of NO donor compounds and antioxidants for 8, 12, and 20 hours. Apoptosis of neutrophils was determined in vitro by flow cytometric analysis of cellular DNA content and Annexin V protein binding to the cell surface. Exposure of human neutrophils to GEA-3162 and SIN-1 significantly accelerates and enhances their apoptosis in vitro in a time-dependent fashion. In the presence of SNP, intensification of apoptosis has not been revealed until 12 hours after the culture. The inhibition of GEA-3162- and SIN-1-mediated neutrophil apoptosis by superoxide dismutase (SOD but not by catalase (CAT was observed. Our results show that SOD and CAT can protect neutrophils against NO-donors-induced apoptosis and suggest that the interaction of NO and oxygen metabolites signals may determine the destructive or protective role of NO donor compounds during apoptotic neutrophil death.

  2. Flow cytometric analysis of platelet cyclooxygenase-1 and -2 and surface glycoproteins in patients with immune thrombocytopenia and healthy individuals.

    Science.gov (United States)

    Rubak, Peter; Kristensen, Steen D; Hvas, Anne-Mette

    2017-06-01

    Immature platelets may contain more platelet enzymes such as cyclooxygenase (COX)-1 and COX-2 than mature platelets. Patients with immune thrombocytopenia (ITP) have a higher fraction of immature platelets and can therefore be utilized as a biological model for investigating COX-1 and COX-2 platelet expression. The aims were to develop flow cytometric assays for platelet COX-1 and COX-2 and to investigate the COX-1 and COX-2 platelet expression, platelet turnover, and platelet glycoproteins in ITP patients (n = 10) compared with healthy individuals (n = 30). Platelet count and platelet turnover parameters (mean platelet volume (MPV), immature platelet fraction (IPF), and immature platelet count (IPC)) were measured by flow cytometry (Sysmex XE-5000). Platelet COX-1, COX-2, and the glycoproteins (GP)IIb, IX, Ib, Ia, and IIIa were all analyzed by flow cytometry (Navios) and expressed as median fluorescence intensity. COX analyses were performed in both whole blood and platelet rich plasma (PRP), whereas platelet glycoproteins were analyzed in whole blood only. ITP patients had significantly lower platelet count (55 × 10 9 /L) than healthy individuals (240 × 10 9 /L, p platelet count and IPC (both p-values Platelet COX-1 expression was higher in ITP patients than healthy individuals using whole blood (p COX-1 platelet turnover and COX-1 expression (all p-values platelet turnover and COX-1 and COX-2 expressions (all p-values platelet turnover in ITP patients (all p-values 0.14, rho = 0.11-0.28). In conclusion, ITP patients expressed higher COX-1 and platelet glycoprotein levels than healthy individuals. COX-1 and platelet glycoproteins demonstrated positive correlations with platelet turnover in ITP patients. In healthy individuals, COX-1 and COX-2 expression correlated positively with platelet turnover. PRP was more sensitive compared with whole blood as regards determination of COX. Therefore, PRP is the recommended matrix for investigating COX-1 and COX-2 in

  3. A flow cytometric assay to quantify invasion of red blood cells by rodent Plasmodium parasites in vivo.

    Science.gov (United States)

    Lelliott, Patrick M; Lampkin, Shelley; McMorran, Brendan J; Foote, Simon J; Burgio, Gaetan

    2014-03-17

    Malaria treatments are becoming less effective due to the rapid spread of drug resistant parasites. Increased understanding of the host/parasite interaction is crucial in order to develop treatments that will be less prone to resistance. Parasite invasion of the red blood cell (RBC) is a critical aspect of the parasite life cycle and is, therefore, a promising target for the development of malaria treatments. Assays for analysing parasite invasion in vitro have been developed, but no equivalent assays exist for in vivo studies. This article describes a novel flow cytometric in vivo parasite invasion assay. Experiments were conducted with mice infected with erythrocytic stages of Plasmodium chabaudi adami strain DS. Exogenously labelled blood cells were transfused into infected mice at schizogony, and collected blood samples stained and analysed using flow cytometry to specifically detect and measure proportions of labelled RBC containing newly invaded parasites. A combination of antibodies (CD45 and CD71) and fluorescent dyes, Hoechst (DNA) and JC-1 (mitochondrial membrane potential), were used to differentiate parasitized RBCs from uninfected cells, RBCs containing Howell-Jolly bodies, leukocytes and RBC progenitors. Blood cells were treated ex vivo with proteases to examine the effects on in vivo parasite invasion. The staining and flow cytometry analysis method was accurate in determining the parasitaemia down to 0.013% with the limit of detection at 0.007%. Transfused labelled blood supported normal rates of parasite invasion. Protease-treated red cells resulted in 35% decrease in the rate of parasite invasion within 30 minutes of introduction into the bloodstream of infected mice. The invasion assay presented here is a versatile method for the study of in vivo red cell invasion efficiency of Plasmodium parasites in mice, and allows direct comparison of invasion in red cells derived from two different populations. The method also serves as an accurate

  4. Microbial Eco-Physiology of the human intestinal tract: a flow cytometric approach

    NARCIS (Netherlands)

    Amor, Ben K.

    2004-01-01

    This thesis describes a multifaceted approach to further enhance our view of the complex human intestinal microbial ecosystem. This approach combines me advantages of flow cyrometry (FCM), a single cell and high-throughput technology, and molecular techniques that have proven themselves to be

  5. Flow cytometric characterization of peripheral blood leukocyte populations of 3 neotropical snake species: Boa constrictor, Bothrops jararaca, and Crotalus durissus.

    Science.gov (United States)

    Carvalho, Marcelo P N; Queiroz-Hazarbassanov, Nicolle G T; Massoco, Cristina O; Rossi, Silmara; Sant'Anna, Sávio S; Catão-Dias, José L; Grego, Kathleen F

    2016-06-01

    The reptilian immune system is represented by innate, humoral, and cell-mediated mechanisms, involving different types of blood leukocytes. The development of optimized methods for the advanced study of origin and function of reptilian blood leukocytes is needed. The purpose of the study was to optimize leukocyte density gradient isolation protocols from snake peripheral blood samples, and characterize recovered cells by flow cytometry based on size and internal complexity for a qualitative and semi-quantitative assessment of leukocyte populations in one boa (Boa constrictor), and 2 viper species (Bothrops jararaca, Crotalus durissus). Blood samples from 30 snakes (10 from each species, 5 males and 5 females) were collected in tubes with sodium heparin. Fresh blood was centrifuged with either ficoll-paque PLUS or percoll density gradients for leukocyte isolation. Flow cytometric leukocyte gates were defined based on size (forward scatter [FSC]) and internal complexity (side scatter [SSC]). Relative leukocyte differential counts after sorting the cells in these gates in one snake for each species were compared to conventional light microscopic differential counts on unsorted isolated leukocytes. There was no statistical difference in the relative leukocyte populations, including heterophils, azurophils, and small and large lymphocytes between samples isolated by ficoll or percoll. Four leukocyte gates were identified based on their location in FSC/SSC cytograms. The relative leukocyte differential counts after sorting in single animals showed some agreement with the light microscopy differential count on unsorted cells. Based on FSC and SSC, 4 distinct leukocyte populations were found in ficoll or percoll density gradient isolated leukocytes from peripheral blood from boa and viper species. Further optimization of the technique should allow the performance of functional assays. © 2016 American Society for Veterinary Clinical Pathology.

  6. Flow cytometric gating for spleen monocyte and DC subsets: differences in autoimmune NOD mice and with acute inflammation.

    Science.gov (United States)

    Dong, Matthew B; Rahman, M Jubayer; Tarbell, Kristin V

    2016-05-01

    The role of antigen presenting cells (APCs) in the pathogenesis of autoimmune and other inflammatory diseases is now better understood due to advances in multicolor flow cytometry, gene expression analysis of APC populations, and functional correlation of mouse to human APC populations. A simple but informative nomenclature of conventional and plasmacytoid dendritic cell subsets (cDC1, cDC2, pDC) and monocyte-derived populations incorporates these advances, but accurate subset identification is critical. Ambiguous gating schemes and alterations of cell surface markers in inflammatory condition can make comparing results between studies difficult. Both acute inflammation, such as TLR-ligand stimulation, and chronic inflammation as found in mouse models of autoimmunity can alter DC subset gating. Here, we address these issues using in vivo CpG stimulation as an example of acute inflammation and the non-obese diabetic (NOD) mouse as a model of chronic inflammation.We provide a flow cytometric antibody panel and gating scheme that differentiate 2 monocytic and 3DC subsets in the spleen both at steady state and after CpG stimulation. Using this method, we observed differences in the composition of NOD DCs that have been previously reported, and newly identified increases in the number of NOD monocyte-derived DCs. Finally, we established a protocol for DC phosphoflow to measure the phosphorylation state of intracellular proteins, and use it to confirm functional differences in the identified subsets. Therefore, we present optimized methods for distinguishing monocytic and DC populations with and without inflammation and/or autoimmunity associated with NOD mice. Published by Elsevier B.V.

  7. Single-laboratory validation of a multiplex flow cytometric immunoassay for the simultaneous detection of coccidiostats in eggs and feed.

    Science.gov (United States)

    Bienenmann-Ploum, Monique E; Vincent, Ursula; Campbell, Katrina; Huet, Anne-Catherine; Haasnoot, Willem; Delahaut, Philippe; Stolker, Linda A A M; Elliott, Christopher T; Nielen, Michel W F

    2013-11-01

    Coccidiostats are authorized in the European Union (EU) to be used as poultry feed additives. Maximum (residue) levels (M(R)Ls) have been set within the EU for consumer and animal protection against unintended carry-over, and monitoring is compulsory. This paper describes the single-laboratory validation of a previously developed multiplex flow cytometric immunoassay (FCIA) as screening method for coccidiostats in eggs and feed and provides and compares different approaches for the calculation of the cut-off levels which are not described in detail within Commission Decision 2002/657/EC. Comparable results were obtained between the statistical (reference) approach and the rapid approaches. With the most rapid approach, the cut-off levels for narasin/salinomycin, lasalocid, diclazuril, nicarbazin (DNC) and monensin in egg, calculated as percentages of inhibition (%B/B0), were 60, 32, 76, 80 and 84, respectively. In feed, the cut-off levels for narasin/salinomycin, lasalocid, nicarbazin (DNC) and monensin were 70, 64, 72 and 78, respectively, and could not be determined for diclazuril. For all analytes, except for diclazuril in feed, the rate of false positives (false non-compliant) in blank samples was lower than 1 %, and the rate of false negatives (false compliant) at the M(R)Ls was below 5 %. Additionally, very good correlations (r ranging from 0.994 to 0.9994) were observed between two different analysers, a sophisticated flow cytometer (FlexMAP 3D(®)) and a more cost-efficient and transportable planar imaging detector (MAGPIX(®)), hence demonstrating adequate transferability.

  8. Flow cytometric evaluation of sperm parameters in relation to fertility potential.

    Science.gov (United States)

    Gillan, Lindsay; Evans, Gareth; Maxwell, W M C

    2005-01-15

    Most laboratory methods used to evaluate semen quality have not correlated highly with fertilizing capacity. The discovery of a variety of fluorochromes and compounds conjugated to fluorescent probes has enabled a more widespread analysis of sperm attributes, and in conjunction with the flow cytometer, permit the evaluation of a large number of spermatozoa. A number of characteristics of sperm integrity, viability and function can be assessed by flow cytometry. The DNA status of spermatozoa has been determined using the metachromatic properties of acridine orange (AO). AO staining, when used in the sperm chromatin structure assay (SCSA), correlates with fertility in a number of species. DNA fragmentation can also be assessed using the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay, which identifies DNA strand breaks by labeling free 3'-OH termini with modified nucleotides. The status of the sperm acrosome can be determined using fluorescently labeled lectins and LysoTracker Green DND-26, a fluorescent acidotropic probe. Capacitation status has been observed through calcium-mediated changes using chlortetracycline (CTC) or by changes in membrane fluidity monitored by the binding of the fluorescent amphiphilic probe, Merocyanine 540. Fluorescently labeled annexin-V, C6NBD and Ro-09-0198 can also be used to detect changes in membrane phospholipid distribution. Cell viability can be determined using the propensity of propidium iodide (PI), ethidium homodimer-1 (EthD-1) or Yo-Pro-1 to permeate damaged membranes. These are generally more adaptable to clinical flow cytometry than the bisbenzimide membrane impermeable stain, Hoechst 33258, which excites in the ultraviolet range and requires UV laser equipment. Mitochondrial function can be determined using rhodamine 123 (R123) and MitoTracker Green FM (MITO) and 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolyl-carbocyanine iodide (JC-1). Flow cytometry is a tool that may be used

  9. Flow cytometric determination of osmotic behaviour of animal erythrocytes toward their engineering for drug delivery

    Directory of Open Access Journals (Sweden)

    Kostić Ivana T.

    2015-01-01

    Full Text Available Despite the fact that the methods based on the osmotic properties of the cells are the most widely used for loading of drugs in human and animal erythrocytes, data related to the osmotic properties of erythrocytes derived from animal blood are scarce. This work was performed with an aim to investigate the possibility of use the flow cytometry as a tool for determination the osmotic behaviour of porcine and bovine erythrocytes, and thus facilitate the engineering of erythrocytes from animal blood to be drug carriers. The method of flow cytometry successfully provided the information about bovine and porcine erythrocyte osmotic fragility, and made the initial steps in assessment of erythrocyte shape in a large number of erythrocytes. Although this method is not able to confirm the swelling of pig erythrocytes, it indicated to the differences in pig erythrocytes that had basic hematological parameters inside and outside the reference values. In order to apply/use the porcine and bovine erythrocytes as drug carriers, the method of flow cytometry, confirming the presence of osmotically different fractions of red blood cells, indicated that various amounts of the encapsulated drug in porcine and bovine erythrocytes can be expected.

  10. Multiparameter Flow Cytometric Assays to Quantify Effector and Regulatory T-Cell Function in Multiple Sclerosis.

    Science.gov (United States)

    Sinha, Sushmita; Crawford, Michael P; Ortega, Sterling B; Karandikar, Nitin J

    2015-01-01

    The immune system plays a major pathological and regulatory role in multiple sclerosis (MS) and, therefore, is a focus of extensive research. Animal models of MS have been crucial in understanding the pathological processes in MS and developing certain treatments, however, all crucial aspects of the human disease may not be appropriately modeled. With the exception of detecting oligoclonal bands and IgG synthesis in cerebrospinal fluids of MS patients, there has not been major progress in the development of immunologic tests that can be used for diagnosis of MS. Further, due to the lack of validated immune assays, routine monitoring of the immune system following therapy initiation is not a part of standard patient care in MS. This is critical since immunomodulatory therapies used for MS treatment are not benign and, more importantly, there is a considerable variation in clinical responses in MS patients initiating such therapies. Flow cytometry is a powerful tool that can be used for studying both the phenotype and function of immune cells. The studies described here will demonstrate how flow cytometry can be used to apply current knowledge about the MS immune system to develop a diagnostic laboratory test for the immunologic monitoring of this disease. Importantly, we will also show that the multiparameter flow cytometry based assay developed by us can also be implemented for the immunologic evaluation of therapeutic success in MS patients.

  11. A flow cytometric method for characterization of circulating cell-derived microparticles in plasma.

    Science.gov (United States)

    Nielsen, Morten Hjuler; Beck-Nielsen, Henning; Andersen, Morten Nørgaard; Handberg, Aase

    2014-01-01

    Previous studies on circulating microparticles (MPs) indicate that the majority of MPs are of a size below the detection limit of most standard flow cytometers. The objective of the present study was to establish a method to analyze MP subpopulations above the threshold of detection of a new generation BD FACSAria™ III digital flow cytometer. We analyzed MP subpopulations in plasma from 24 healthy individuals (9 males and 15 females). MPs were identified according to their size (derived from platelets, monocytes, erythrocytes and endothelial cells. In addition, levels of tissue factor-positive MPs were determined. The FACSAria demonstrated improved sensitivity and increased MP detection range compared to the FC500 instrument. The reproducibility of PS+PMP and PS+MP measurements was 11.7 and 23.2%, respectively. When expressed as a percentage of total MPs, the PS-positive MP population represented 15.1±5.5%, and PS-positive MPs were significantly increased in men. We have established a method to measure MPs above the detection limit of a new generation flow cytometer and derived from a number of cell-types in a healthy population of men and women.

  12. Simple flow cytometric detection of haemozoin containing leukocytes and erythrocytes for research on diagnosis, immunology and drug sensitivity testing

    Directory of Open Access Journals (Sweden)

    Grobusch Martin P

    2011-03-01

    Full Text Available Abstract Background Malaria pigment (haemozoin, Hz has been the focus of diverse research efforts. However, identification of Hz-containing leukocytes or parasitized erythrocytes is usually based on microscopy, with inherent limitations. Flow cytometric detection of depolarized Side-Scatter is more accurate and its adaptation to common bench top flow cytometers might allow several applications. These can range from the ex-vivo and in-vitro detection and functional analysis of Hz-containing leukocytes to the detection of parasitized Red-Blood-Cells (pRBCs to assess antimalarial activity. Methods A standard benchtop flow cytometer was adapted to detect depolarized Side-Scatter. Synthetic and Plasmodium falciparum Hz were incubated with whole blood and PBMCs to detect Hz-containing leukocytes and CD16 expression on monocytes. C5BL/6 mice were infected with Plasmodium berghei ANKA or P. berghei NK65 and Hz-containing leukocytes were analysed using CD11b and Gr1 expression. Parasitized RBC from infected mice were identified using anti-Ter119 and SYBR green I and were analysed for depolarized Side Scatter. A highly depolarizing RBC population was monitored in an in-vitro culture incubated with chloroquine or quinine. Results A flow cytometer can be easily adapted to detect depolarized Side-Scatter and thus, intracellular Hz. The detection and counting of Hz containing leukocytes in fresh human or mouse blood, as well as in leukocytes from in-vitro experiments was rapid and easy. Analysis of CD14/CD16 and CD11b/Gr1 monocyte expression in human or mouse blood, in a mixed populations of Hz-containing and non-containing monocytes, appears to show distinct patterns in both types of cells. Hz-containing pRBC and different maturation stages could be detected in blood from infected mice. The analysis of a highly depolarizing population that contained mature pRBC allowed to assess the effect of chloroquine and quinine after only 2 and 4 hours, respectively

  13. Flow Cytometric Detection of PrPScin Neurons and Glial Cells from Prion-Infected Mouse Brains.

    Science.gov (United States)

    Yamasaki, Takeshi; Suzuki, Akio; Hasebe, Rie; Horiuchi, Motohiro

    2018-01-01

    In prion diseases, an abnormal isoform of prion protein (PrP Sc ) accumulates in neurons, astrocytes, and microglia in the brains of animals affected by prions. Detailed analyses of PrP Sc -positive neurons and glial cells are required to clarify their pathophysiological roles in the disease. Here, we report a novel method for the detection of PrP Sc in neurons and glial cells from the brains of prion-infected mice by flow cytometry using PrP Sc -specific staining with monoclonal antibody (MAb) 132. The combination of PrP Sc staining and immunolabeling of neural cell markers clearly distinguished neurons, astrocytes, and microglia that were positive for PrP Sc from those that were PrP Sc negative. The flow cytometric analysis of PrP Sc revealed the appearance of PrP Sc -positive neurons, astrocytes, and microglia at 60 days after intracerebral prion inoculation, suggesting the presence of PrP Sc in the glial cells, as well as in neurons, from an early stage of infection. Moreover, the kinetic analysis of PrP Sc revealed a continuous increase in the proportion of PrP Sc -positive cells for all cell types with disease progression. Finally, we applied this method to isolate neurons, astrocytes, and microglia positive for PrP Sc from a prion-infected mouse brain by florescence-activated cell sorting. The method described here enables comprehensive analyses specific to PrP Sc -positive neurons, astrocytes, and microglia that will contribute to the understanding of the pathophysiological roles of neurons and glial cells in PrP Sc -associated pathogenesis. IMPORTANCE Although formation of PrP Sc in neurons is associated closely with neurodegeneration in prion diseases, the mechanism of neurodegeneration is not understood completely. On the other hand, recent studies proposed the important roles of glial cells in PrP Sc -associated pathogenesis, such as the intracerebral spread of PrP Sc and clearance of PrP Sc from the brain. Despite the great need for detailed analyses

  14. Flow cytometric analysis of apoptosis in cryoconserved chicken primordial germ cells.

    Science.gov (United States)

    Sawicka, Dorota; Chojnacka-Puchta, Luiza; Zielinski, Marcin; Plucienniczak, Grazyna; Plucienniczak, Andrzej; Bednarczyk, Marek

    2015-03-01

    Our research aimed to compare the effects of four cryoprotectants and four slow freezing programs on the viability and apoptosis of primordial germ cells (PGCs) in vitro. PGCs were collected from chicken embryonic blood at Hamburger and Hamilton (HH) stages 14-16 and purified by Percoll density gradient centrifugation and then subjected to cryopreservation. We applied microscopy to determine the survival of PGCs after trypan blue staining and flow cytometry to examine apoptosis and viability after annexin V kit staining. We also examined the functionality of cryopreserved PGCs in vivo. Significant differences in viability of PGCs determined via microscopy and flow cytometry were observed. The most unfavorable combination for slow freezing PGCs was program 3 and MIX H (10% DMSO and 5% glycerol in Hank's solution supplemented with 10% FBS) as the cryoprotectant (48.43 and 15.37% live and early apoptotic PGCs, respectively). The highest average percentage of live PGCs (93.1%) and the lowest percentage of early apoptotic PGCs (6.5%) were achieved by slow freezing PGCs in the presence of DMSO F (10% DMSO in FBS) via program 1. Therefore, this method was chosen for the in vivo test. Cryopreserved (group 1) and freshly isolated (group 2) PGCs were transfectedwith a pEGFP-N1 plasmid, cultured under antibiotic selection, and then injected into 3-day-old embryos. After 5 days of incubation, we identified the EGFP marker gene in the gonads of 40 and 45% of recipients in groups 1 and 2, respectively. This is the first study to apply flow cytometry to examine the apoptosis and viability of cryopreserved PGCs. The in vitro and in vivo findings showed that the developed PGC cryoconservation method, depending on slow freezing at the rate of 2°C/min (program 1) in the presence of 10% DMSO F, is an improvement over previous cryoconservation methods and may be a useful tool for the ex situ strategy of poultry biodiversity preservation.

  15. Quick cytogenetic screening of breeding bulls using flow cytometric sperm DNA histogram analysis.

    Science.gov (United States)

    Nagy, Szabolcs; Polgár, Péter J; Andersson, Magnus; Kovács, András

    2016-09-01

    The aim of the present study was to test the FXCycle PI/RNase kit for routine DNA analyses in order to detect breeding bulls and/or insemination doses carrying cytogenetic aberrations. In a series of experiments we first established basic DNA histogram parameters of cytogenetically healthy breeding bulls by measuring the intraspecific genome size variation of three animals, then we compared the histogram profiles of bulls carrying cytogenetic defects to the baseline values. With the exception of one case the test was able to identify bulls with cytogenetic defects. Therefore, we conclude that the assay could be incorporated into the laboratory routine where flow cytometry is applied for semen quality control.

  16. APPLICATION OF SIX-COLOR FLOW CYTOMETRIC ANALYSIS FOR IMMUNE PROFILE MONITORING

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    I. V. Kudryavtsev

    2015-01-01

    Full Text Available The article deals with applications of six-color flow analysis for studying the immune state parameters by means of flow cytometry. Whole blood from healthy donors was stained with combination of monoclonal antibodies, i.e., HLA-DR-FITC, CD16+56-PE, CD4-ECD, CD19-ECD, CD8-PC5.5, CD3-PC7, CD45-APC (Beckman Coulter, USA using a “no-wash” technology. To adjust the photomultiplier (PMT voltage, we used the tubes with each of the tested monoclonal antibodies, PMT voltage was considered optimal when the negative population was located in the middle of the first decade at the logarithmic scale. The compensatory adjustment was performed in automatic mode, using Navios software (Beckman Coulter, USA. We discuss an optimal gating strategy in order to assess the populations of interest: total T cells (CD3+CD19-, T helper cells (CD3+CD4+, cytotoxic T lymphocytes (CD3+CD8+, B cells (CD3-CD19+, NK cells (CD3-CD16+CD56+, NKT cells (CD3+CD16+CD56+, activated T lymphocytes (CD3+HLA-DR+. 

  17. Optimization of flow cytometric detection and cell sorting of transgenic Plasmodium parasites using interchangeable optical filters.

    Science.gov (United States)

    Vorobjev, Ivan A; Buchholz, Kathrin; Prabhat, Prashant; Ketman, Kenneth; Egan, Elizabeth S; Marti, Matthias; Duraisingh, Manoj T; Barteneva, Natasha S

    2012-09-05

    Malaria remains a major cause of morbidity and mortality worldwide. Flow cytometry-based assays that take advantage of fluorescent protein (FP)-expressing malaria parasites have proven to be valuable tools for quantification and sorting of specific subpopulations of parasite-infected red blood cells. However, identification of rare subpopulations of parasites using green fluorescent protein (GFP) labelling is complicated by autofluorescence (AF) of red blood cells and low signal from transgenic parasites. It has been suggested that cell sorting yield could be improved by using filters that precisely match the emission spectrum of GFP. Detection of transgenic Plasmodium falciparum parasites expressing either tdTomato or GFP was performed using a flow cytometer with interchangeable optical filters. Parasitaemia was evaluated using different optical filters and, after optimization of optics, the GFP-expressing parasites were sorted and analysed by microscopy after cytospin preparation and by imaging cytometry. A new approach to evaluate filter performance in flow cytometry using two-dimensional dot blot was developed. By selecting optical filters with narrow bandpass (BP) and maximum position of filter emission close to GFP maximum emission in the FL1 channel (510/20, 512/20 and 517/20; dichroics 502LP and 466LP), AF was markedly decreased and signal-background improve dramatically. Sorting of GFP-expressing parasite populations in infected red blood cells at 90 or 95% purity with these filters resulted in 50-150% increased yield when compared to the standard filter set-up. The purity of the sorted population was confirmed using imaging cytometry and microscopy of cytospin preparations of sorted red blood cells infected with transgenic malaria parasites. Filter optimization is particularly important for applications where the FP signal and percentage of positive events are relatively low, such as analysis of parasite-infected samples with in the intention of gene

  18. Optimization of flow cytometric detection and cell sorting of transgenic Plasmodium parasites using interchangeable optical filters

    Directory of Open Access Journals (Sweden)

    Vorobjev Ivan A

    2012-09-01

    Full Text Available Abstract Background Malaria remains a major cause of morbidity and mortality worldwide. Flow cytometry-based assays that take advantage of fluorescent protein (FP-expressing malaria parasites have proven to be valuable tools for quantification and sorting of specific subpopulations of parasite-infected red blood cells. However, identification of rare subpopulations of parasites using green fluorescent protein (GFP labelling is complicated by autofluorescence (AF of red blood cells and low signal from transgenic parasites. It has been suggested that cell sorting yield could be improved by using filters that precisely match the emission spectrum of GFP. Methods Detection of transgenic Plasmodium falciparum parasites expressing either tdTomato or GFP was performed using a flow cytometer with interchangeable optical filters. Parasitaemia was evaluated using different optical filters and, after optimization of optics, the GFP-expressing parasites were sorted and analysed by microscopy after cytospin preparation and by imaging cytometry. Results A new approach to evaluate filter performance in flow cytometry using two-dimensional dot blot was developed. By selecting optical filters with narrow bandpass (BP and maximum position of filter emission close to GFP maximum emission in the FL1 channel (510/20, 512/20 and 517/20; dichroics 502LP and 466LP, AF was markedly decreased and signal-background improve dramatically. Sorting of GFP-expressing parasite populations in infected red blood cells at 90 or 95% purity with these filters resulted in 50-150% increased yield when compared to the standard filter set-up. The purity of the sorted population was confirmed using imaging cytometry and microscopy of cytospin preparations of sorted red blood cells infected with transgenic malaria parasites. Discussion Filter optimization is particularly important for applications where the FP signal and percentage of positive events are relatively low, such as analysis

  19. Flow cytometric measurement of the metabolism of benzo [a] pyrene by mouse liver cells in culture

    International Nuclear Information System (INIS)

    Bartholomew, J.C.; Wade, C.G.; Dougherty, K.

    1984-01-01

    The metabolism of benzo[a]pyrene in individual cells was monitored by flow cytometry. The measurements are based on the alterations that occur in the fluorescence emission spectrum of benzo[a]pyrene when it is converted to various metabolities. Using present instrumentation the technique could easily detect 1 x 10/sup 6/ molecules per cells of benzo [a]pyrene and 1 x 10/sup 7/ molecules per cell of the diol epoxide. The analysis of C3H IOT 1/2 mouse fibroblasts growing in culture indicated that there was heterogeneity in the conversion of the parent compound into diol epoxide derivative suggesting that some variation in sensitivity to transformation by benzo[a]pyrene may be due to differences in cellular metabolism

  20. DEA 1 expression on dog erythrocytes analyzed by immunochromatographic and flow cytometric techniques.

    Science.gov (United States)

    Acierno, M M; Raj, K; Giger, U

    2014-01-01

    The Dog erythrocyte antigen (DEA) 1 blood group system was thought to contain types DEA 1.1 and 1.2 (and possibly 1.3 [A3]). However, DEA 1.2+ dogs are very rare and newer typing methods reveal varying degrees of DEA 1 positivity. To assess if variation in DEA 1 positivity is because of quantitative differences in surface antigen expression. To determine expression patterns in dogs over time and effects of blood storage (4°C). To evaluate DEA 1.2+ samples by DEA 1 typing methods. Anticoagulated blood samples from 66 dogs in a research colony and from a hospital, and 9 previously typed DEA 1.2+ dogs from an animal blood bank. Research study: Samples were analyzed by flow cytometry and immunochromatographic strip using a monoclonal anti-DEA 1 antibody. Twenty dogs were DEA 1-, whereas 46 dogs were weakly to strongly DEA 1+. Antigen quantification revealed excellent correlation between strip and flow cytometry (r = 0.929). Both methods reclassified DEA 1.2+ samples as weakly to moderately DEA 1+, but they were not retyped with the polyclonal anti-DEA 1.1/1.X antibodies. Dogs and blood samples retained their relative DEA 1 antigen densities over time. The blood group system DEA 1 is a continuum from negative to strongly positive antigen expression. Previously typed DEA 1.2+ appears to be DEA 1+. These findings further the understanding of the DEA 1 system and suggest that all alleles within the DEA 1 system have a similarly based epitope recognized by the monoclonal antibody. Copyright © 2014 by the American College of Veterinary Internal Medicine.

  1. Flow cytometric analysis of lymphocytes and lymphocyte subpopulations in induced sputum from patients with asthma

    Directory of Open Access Journals (Sweden)

    Yutaro Shiota

    2000-01-01

    Full Text Available Study objectives were to compare the numbers of lymphocytes and lymphocyte subpopulations in induced sputum from asthmatic patients and from healthy subjects, and to determine the effect of inhaled anti-asthmatic steroid therapy on these cell numbers. Hypertonic saline inhalation was used to non-invasively induce sputum samples in 34 patients with bronchial asthma and 21 healthy subjects. The sputum samples were reduced with dithioerythritol and absolute numbers of lymphocytes and lymphocyte subpopulations were assessed by direct immunofluorescence and flow cytometry. To assess the effect of beclomethasone dipropionate (BDP on induced sputum, numbers of lymphocytes and lymphocyte subpopulations in sputum also were evaluated after 4 weeks of BDP inhalation treatment in seven asthmatic patients. An adequate sample was obtained in 85.3% of patients with asthma and in 79.2% of the healthy subjects. Induced sputum from patients with asthma had increased numbers of lymphocytes (P = 0.009; CD4+ cells (P = 0.044; CD4+ cells-bearing interleukin-2 receptor (CD25; P = 0.016; and CD4+ cells bearing human histocompatibility leukocyte antigen (HLA-DR (P = 0.033. CD8+ cells were not increased in asthmatic patients. In patients treated with inhaled steroids, numbers of lymphocytes, CD4+ cells, CD25-bearing CD4+ cells and HLA-DR-bearing CD4+ cells in sputum decreased from pretreatment numbers (P = 0.016, 0.002, 0.003 and 0.002, respectively. Analysis of lymphocytes in induced sputum by flow cytometry is useful in assessing bronchial inflammation, and activated CD4+ lymphocytes may play a key role in the pathogenesis of airway inflammation in bronchial asthma.

  2. Flow cytometric assessment of microbial abundance in the near-field area of seawater reverse osmosis concentrate discharge

    KAUST Repository

    Van Der Merwe, Riaan

    2014-06-01

    The discharge of concentrate and other process waters from seawater reverse osmosis (SWRO) plant operations into the marine environment may adversely affect water quality in the near-field area surrounding the outfall. The main concerns are the increase in salt concentration in receiving waters, which results in a density increase and potential water stratification near the outfall, and possible increases in turbidity, e.g., due to the discharge of filter backwash waters. Changes in ambient water quality may affect microbial abundance in the area, for example by hindering the photosynthesis process or disrupting biogenesis. It is widely accepted that marine biodiversity is lower in more extreme conditions, such as high salinity environments. As aquatic microbial communities respond very rapidly to changes in their environment, they can be used as indicators for monitoring ambient water quality. The objective of this study was to assess possible changes in microbial abundance as a result of concentrate discharge into the near-field area (<. 25. m) surrounding the outfall of the King Abdullah University of Science and Technology (KAUST) SWRO plant. Flow cytometric (FCM) analysis was conducted in order to rapidly determine microbial abundance on a single-cell level in 107 samples, taken by diving, from the discharge area, the intake area and two control sites. FCM analysis combined the measurement of distinct scatter of cells and particles, autofluorescence of cyanobacteria and algae, and fluorescence after staining of nucleic acids with SYBR® Green for a total bacterial count. The results indicate that changes in microbial abundance in the near-field area of the KAUST SWRO outfall are minor and appear to be the result of a dilution effect rather than a direct impact of the concentrate discharge. © 2014 Elsevier B.V.

  3. Flow cytometric bacterial cell counts challenge conventional heterotrophic plate counts for routine microbiological drinking water monitoring.

    Science.gov (United States)

    Van Nevel, S; Koetzsch, S; Proctor, C R; Besmer, M D; Prest, E I; Vrouwenvelder, J S; Knezev, A; Boon, N; Hammes, F

    2017-04-15

    Drinking water utilities and researchers continue to rely on the century-old heterotrophic plate counts (HPC) method for routine assessment of general microbiological water quality. Bacterial cell counting with flow cytometry (FCM) is one of a number of alternative methods that challenge this status quo and provide an opportunity for improved water quality monitoring. After more than a decade of application in drinking water research, FCM methodology is optimised and established for routine application, supported by a considerable amount of data from multiple full-scale studies. Bacterial cell concentrations obtained by FCM enable quantification of the entire bacterial community instead of the minute fraction of cultivable bacteria detected with HPC (typically water samples per day, depending on the laboratory and selected staining procedure(s). Moreover, many studies have shown FCM total (TCC) and intact (ICC) cell concentrations to be reliable and robust process variables, responsive to changes in the bacterial abundance and relevant for characterising and monitoring drinking water treatment and distribution systems. The purpose of this critical review is to initiate a constructive discussion on whether FCM could replace HPC in routine water quality monitoring. We argue that FCM provides a faster, more descriptive and more representative quantification of bacterial abundance in drinking water. Copyright © 2017 Elsevier Ltd. All rights reserved.

  4. Evaluation of chromatin condensation in human spermatozoa: a flow cytometric assay using acridine orange staining.

    Science.gov (United States)

    Golan, R; Shochat, L; Weissenberg, R; Soffer, Y; Marcus, Z; Oschry, Y; Lewin, L M

    1997-01-01

    The quality of sperm chromatin is an important factor in fertilization and is especially critical where one spermatozoon is artificially selected for fertilizing an egg (as in intracytoplasmic sperm injection). In this study, flow cytometry after staining of human spermatozoa with Acridine Orange was used to study chromatin structure. A method is described for estimating the percentage of cells in a human sperm sample that have completed epididymal maturation in regard to chromatin condensation. Of the 121 samples of the semen that were examined, nine contained a higher percentage of hypocondensed spermatozoa and six samples contained elevated amounts of hypercondensed spermatozoa. In addition to aberrancies in chromatin condensation other defects showed up as satellite populations of spermatozoa with higher than normal ratios of red/green fluorescence after Acridine Orange staining. Such defects were found in 15 semen samples. The use of swim-up and Percoll gradient centrifugation methods was shown to improve the percentage of spermatozoa with normal chromatin structure in some samples with poor initial quality.

  5. Comparison of quantitative flow cytometric data provided by panels with lower and increased color number

    Science.gov (United States)

    Bocsi, József; Mittag, Anja; Pierzchalski, Arkadiusz; Baumgartner, Adolf; Dähnert, Ingo; Tárnok, Attila

    2012-03-01

    To date the flow cytometry (FCM) industry is booming with new generations of commercial clinical instruments. Long-term clinical studies have the dilemma that moving to new instruments being capable of more complex cell-analysis makes it difficult to compare new data with those obtained on older instruments with less complex analysis panels. Since 15 years we conduct follow-up studies on children with congenital heart diseases. In this period we moved from 2- to 3- and now to 10-color FCM immunophenotyping panels. Questions arise how to compare and transfer data from lower to higher level of complexity. Two comparable antibody panels for leukocyte immunophenotyping (12-tube 2-colors, and 9-tube 4-colors) were measured on a BD FACScalibur FCM (calibration: Spherotech beads) in 19 blood samples from children with congenital heart disease. This increase of colors was accompanied by moving antibodies that were in the 2-color panel either FITC or PE labeled to red dyes such as PerCP or APC. Algorithms were developed for bridging data for quantitative characterization of antigen expression (mean fluorescence intensity) and frequency of different cell subpopulations in combination with rainbow bead standard data. This approach worked for the most relevant antibodies (CD3, CD4, CD8 etc.) well, but rendered substantial uncertainty for activation markers (CD69 etc.). Our techniques are particularly well suited to the analysis in long-term studies and have the potential to compare older and recent results in a standardized way.

  6. Flow cytometric analysis of X-ray sensitivity in ataxia telangiectasia

    International Nuclear Information System (INIS)

    Rudolph, N.S.; Latt, S.A; Harvard Medical School, Boston

    1989-01-01

    Flow cytrometric analysis of 5-bromodeoxyuridine (BrdU) incorporation during DNA synthesis was used to characterize the effects of X-rays on cell-cycle kinetics in the DNA-repair deficiency disease ataxia telangiectasia (AT). Cultured fibroblasts from homozygotes (at/at), heterozygotes (at/+) and normal controls (+/+) were either: (1) irradiated, cultured, then pulsed with BrdU and harvested, or (2) pulsed with BrdU, irradiated, cultured and then harvested. Cells were then fixed and stained with both a fluoresceinated monoclonal antibody against BrdU to identify S-phase cells and with propidium diiodide to measure total DNA content. Irradiation of +/+ and at/+ cells induced a similar, transient G 2 /M arrest detectable within 8 h, which subsequently delayed by 6-8 h the passage of cells into G 1 and depleted early S phase. In contrast, at/at cells failed to arrest in G 2 /M phase and entered the next cell cycle withou pausing to repair radiation-induced damage. X-Rays also blocked entry of +/+ G 1 cells into S phase, subsequently reducing the total S-phase population. This effect was not observed in at/at cells. These cell-cycle responses to radiation may be of diagnostic use and ultimately may help explain the basic defect in AT. (author). 38 refs.; 7 figs.; 2 tabs

  7. Flow cytometric investigation of immune-response-related surface molecules on human colorectal cancers

    DEFF Research Database (Denmark)

    Diederichsen, Axel Cosmus Pyndt; Stenholm, A C; Kronborg, O

    1998-01-01

    different colorectal cancers were monitored by multiparameter flow cytometry. Gating on EP4+ cells, the expression of the surface molecules HLA class I, HLA class II, CD80 (B7-1), CD54 (ICAM-I) and CD58 (LFA-3) was evaluated. In 60 of 70 tumours, all tumour cells expressed HLA class I, in 10 tumours 15......Our purpose was to clarify whether human colorectal cancer cells are equipped to present tumour-associated-antigens to the immune system, and whether this ability correlates with lymphoid infiltration, the Dukes' stage and Jass classification. Enzymatically dissociated tumour cells from 70......-96% of the tumour cells expressed HLA class I. In 1 tumour, all tumour cells expressed HLA class II, in 67 tumours some expressed HLA class II, in 2 tumours none expressed HLA class II. Expression of CD58 was heterogeneous, and there was no or only sparse expression of CD80 and CD54. Expression of the HLA class I...

  8. Flow cytometric analysis of microparticle phenotype and their role in thrombin generation.

    Science.gov (United States)

    Macey, M G; Enniks, N; Bevan, S

    2011-01-01

    Microparticles may be generated from a number of cell types and are known to play a role in haemostasis by a variety of mechanisms. We investigated the role of platelet, red cell, and leucocyte-derived microparticles in the measurement of thrombin generation. Four parameters of thrombin generation (the endogenous thrombin potential (ETP), lag time, time to peak, peak height) and microparticle content was determined in 35 plasma samples from normal individuals pre and post filtration to remove microparticles. Immunofluorescent flow cytometry was used to identify and enumerate platelet, leucocyte, monocyte and red cell derived microparticles in plasma samples based on the expression of CD42b, CD45, CD15, and Glycophorin A respectively. Expression of phosphatidylserine and tissue factor by microparticles was determined by Annexin V and anti CD142 binding. The pre and post filtration results were compared. There was a significant decrease in ETP and Peak Height, and an increase in the time to peak post filtration (P microparticles was also observed. The change in CD42b+ microparticles correlated highly with the change in Annexin V+ microparticles (r = 0.68). Whilst the change in ETP correlated best with the change in CD15+ microparticles (r = 0.45) and the change in time to peak correlated with the change in Annexin V binding (r = 0.52) (P < 0.01). The presence of micropartcles in plasma significantly affects thrombin generation. Copyright © 2010 International Clinical Cytometry Society.

  9. Flow cytometric studies of the survival of cytochalasin-induced polyploidy in Chinese hamster ovary cells.

    Science.gov (United States)

    Court, J B; Burn, C; Moore, J L

    1990-05-01

    A method has been developed to estimate the post-irradiation survival of cytochalasin B-induced polyploidization of adherent Chinese hamster ovary cell using the flow cytometer. After exposure to radiation, surviving cells are allowed to become polyploid in the presence of cytochalasin, and are detached using trypsin, fixed by the addition of glutaraldehyde and stained using mithramycin. DNA content distributions are polymodal, and the absolute number of cells per culture in any given ploidy class is estimated by reference to a non-fluorescent bead internal standard, detected using forward scatter. Post-irradiation survival is defined as the ability to reach a given DNA content, and is reduced exponentially with dose. A bioassay to determine optimum cytochalasin concentrations can be derived from the relative size of the 2C (G0/G1) peak in the DNA content distribution. At culture densities greater than about 8 x 10(4) cell/cm2 the relative number of cells reaching at least 16C is reduced, but this inhibition is partially reversible by an increase in the medium glucose concentration, but not by the use of cytochalasin D or dihydro B.

  10. Proposed criterion for distinguishing ABO mosaics from ABO chimeras using flow cytometric analysis.

    Science.gov (United States)

    Oda, Akira; Matsuyama, Nobuki; Hirashima, Mizuko; Ishii, Hiroyuki; Kimura, Keiko; Matsukura, Harumichi; Hirayama, Fumiya; Kawa, Keisei; Fukumori, Yasuo

    2015-01-01

    Differentiation of ABO mosaics from chimeras is performed using flow cytometry (FCM) analysis. Although mosaics and chimeras have been distinguished by presence or absence of clear resolution using FCM analysis, the lack of quantitative metrics and definitive criteria for this differentiation has made some cases difficult to differentiate. In this study, therefore, we attempted to establish a definitive and quantitative criterion for this differentiation. When FCM histogram gates for group "A" or "B" antigen-negative and -positive red blood cells (RBCs) were set such that group O RBCs were classified as 99 percent negative and group A or B RBCs as 99 percent positive, the percentages of RBCs in the middle region of six chimeras and 23 mosaics (12 A mosaics and 11 B mosaics) were 0.1-0.6 percent and 7.0-19.0 percent, respectively. This results suggested that ABO mosaics and chimeras can be unambiguously differentiated when the cutoff point of the intermediate region is set to 1 percent.

  11. Chemosensitivity of human small cell carcinoma of the lung detected by flow cytometric DNA analysis of drug-induced cell cycle perturbations in vitro

    DEFF Research Database (Denmark)

    Engelholm, S A; Spang-Thomsen, M; Vindeløv, L L

    1986-01-01

    A method based on detection of drug-induced cell cycle perturbation by flow cytometric DNA analysis has previously been described in Ehrlich ascites tumors as a way to estimate chemosensitivity. The method is extended to test human small-cell carcinoma of the lung. Three tumors with different...... sensitivities to melphalan in nude mice were used. Tumors were disaggregated by a combined mechanical and enzymatic method and thereafter have incubated with different doses of melphalan. After incubation the cells were plated in vitro on agar, and drug induced cell cycle changes were monitored by flow...

  12. Monitoring microbiological changes in drinking water systems using a fast and reproducible flow cytometric method

    KAUST Repository

    Prest, Emmanuelle I E C

    2013-12-01

    Flow cytometry (FCM) is a rapid, cultivation-independent tool to assess and evaluate bacteriological quality and biological stability of water. Here we demonstrate that a stringent, reproducible staining protocol combined with fixed FCM operational and gating settings is essential for reliable quantification of bacteria and detection of changes in aquatic bacterial communities. Triplicate measurements of diverse water samples with this protocol typically showed relative standard deviation values and 95% confidence interval values below 2.5% on all the main FCM parameters. We propose a straightforward and instrument-independent method for the characterization of water samples based on the combination of bacterial cell concentration and fluorescence distribution. Analysis of the fluorescence distribution (or so-called fluorescence fingerprint) was accomplished firstly through a direct comparison of the raw FCM data and subsequently simplified by quantifying the percentage of large and brightly fluorescent high nucleic acid (HNA) content bacteria in each sample. Our approach enables fast differentiation of dissimilar bacterial communities (less than 15min from sampling to final result), and allows accurate detection of even small changes in aquatic environments (detection above 3% change). Demonstrative studies on (a) indigenous bacterial growth in water, (b) contamination of drinking water with wastewater, (c) household drinking water stagnation and (d) mixing of two drinking water types, univocally showed that this FCM approach enables detection and quantification of relevant bacterial water quality changes with high sensitivity. This approach has the potential to be used as a new tool for application in the drinking water field, e.g. for rapid screening of the microbial water quality and stability during water treatment and distribution in networks and premise plumbing. © 2013 Elsevier Ltd.

  13. Flow-cytometric study of vital cellular functions in Escherichia coli during solar disinfection (SODIS).

    Science.gov (United States)

    Berney, Michael; Weilenmann, Hans-Ulrich; Egli, Thomas

    2006-06-01

    The effectiveness of solar disinfection (SODIS), a low-cost household water treatment method for developing countries, was investigated with flow cytometry and viability stains for the enteric bacterium Escherichia coli. A better understanding of the process of injury or death of E. coli during SODIS could be gained by investigating six different cellular functions, namely: efflux pump activity (Syto 9 plus ethidium bromide), membrane potential [bis-(1,3-dibutylbarbituric acid)trimethine oxonol; DiBAC4(3)], membrane integrity (LIVE/DEAD BacLight), glucose uptake activity (2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-d-glucose; 2-NBDG), total ATP concentration (BacTiter-Glo) and culturability (pour-plate method). These variables were measured in E. coli K-12 MG1655 cells that were exposed to either sunlight or artificial UVA light. The inactivation pattern of cellular functions was very similar for both light sources. A UVA light dose (fluence) of 80 % of the cells was observed at a fluence of approximately 1500 kJ m(-2), and the cytoplasmic membrane of bacterial cells became permeable at a fluence of >2500 kJ m(-2). Culturable counts of stressed bacteria after anaerobic incubation on sodium pyruvate-supplemented tryptic soy agar closely correlated with the loss of membrane potential. The results strongly suggest that cells exposed to >1500 kJ m(-2) solar UVA (corresponding to 530 W m(-2) global sunlight intensity for 6 h) were no longer able to repair the damage and recover. Our study confirms the lethal effect of SODIS with cultivation-independent methods and gives a detailed picture of the 'agony' of E. coli when it is stressed with sunlight.

  14. Flow cytometric assay for analysis of cytotoxic effects of potential drugs on human peripheral blood leukocytes

    Science.gov (United States)

    Nieschke, Kathleen; Mittag, Anja; Golab, Karolina; Bocsi, Jozsef; Pierzchalski, Arkadiusz; Kamysz, Wojciech; Tarnok, Attila

    2014-03-01

    Toxicity test of new chemicals belongs to the first steps in the drug screening, using different cultured cell lines. However, primary human cells represent the human organism better than cultured tumor derived cell lines. We developed a very gentle toxicity assay for isolation and incubation of human peripheral blood leukocytes (PBL) and tested it using different bioactive oligopeptides (OP). Effects of different PBL isolation methods (red blood cell lysis; Histopaque isolation among others), different incubation tubes (e.g. FACS tubes), anticoagulants and blood sources on PBL viability were tested using propidium iodide-exclusion as viability measure (incubation time: 60 min, 36°C) and flow cytometry. Toxicity concentration and time-depended effects (10-60 min, 36 °C, 0-100 μg /ml of OP) on human PBL were analyzed. Erythrocyte lysis by hypotonic shock (dH2O) was the fastest PBL isolation method with highest viability (>85%) compared to NH4Cl-Lysis (49%). Density gradient centrifugation led to neutrophil granulocyte cell loss. Heparin anticoagulation resulted in higher viability than EDTA. Conical 1.5 mL and 2 mL micro-reaction tubes (both polypropylene (PP)) had the highest viability (99% and 97%) compared to other tubes, i.e. three types of 5.0 mL round-bottom tubes PP (opaque-60%), PP (blue-62%), Polystyrene (PS-64%). Viability of PBL did not differ between venous and capillary blood. A gentle reproducible preparation and analytical toxicity-assay for human PBL was developed and evaluated. Using our assay toxicity, time-course, dose-dependence and aggregate formation by OP could be clearly differentiated and quantified. This novel assay enables for rapid and cost effective multiparametric toxicological screening and pharmacological testing on primary human PBL and can be adapted to high-throughput-screening.°z

  15. Flow cytometric comparison of the effects of trialkyltins on the murine erythroleukemic cell.

    Science.gov (United States)

    Zucker, R M; Elstein, K H; Easterling, R E; Massaro, E J

    1989-10-02

    Cellular effects of exposure to tributyltin (TBT), triethyltin (TET), or trimethyltin (TMT) were investigated by flow cytometry employing the murine erythroleukemic cell (MELC) as a model cellular system. Cell viability was investigated by the carboxyfluorescein diacetate (CFDA) uptake/propidium iodide (PI) exclusion method: above a critical concentration (exposure for 4 h), which was specific for each of the trialkyltin compounds, the cell becomes permeable to PI, indicating loss of viability. Cellular CF fluorescence (derived from intracellular hydrolysis of CFDA) increased as a function of alkyltin concentration below the critical concentration and decreased as viability decreased above the critical concentration. Relative membrane potential, monitored with a cyanine dye (DiOC6), correlated with viability (PI exclusion), remaining essentially unaltered below the critical concentration and decreasing above it. At/above 1 microM TBT, 5 microM TET, or 100 microM TMT, the cell cycle was blocked in the G2/M phase. The 90 degrees light scatter (a measure of refractive index), axial light loss (a measure of volume), and fluorescein isothiocyanate (FITC) fluorescence (a measure of protein content) of nuclei isolated from trialkyltin-treated MELC by detergent treatment, increased as a function of organotin dose. Fluorescence and interference microscopy revealed increased quantities of residual cytoplasmic tags adherent to the nuclei as a function of organotin dose, apparently resulting from increased cytoplasmic resistance to detergent-mediated solubilization. The effects of the trialkyltins correlated with their lipophilicity (octanol/water coefficient). These data support the hypothesis that fixation (protein denaturation, cross-linking, etc.) is an important mode of organotin cytotoxicity.

  16. Flow Cytometric Analysis of Leishmania Reactive CD4+/CD8+ Lymphocyte Proliferation in Cutaneous Leishmaniasis

    Directory of Open Access Journals (Sweden)

    H Keshavarz

    2008-12-01

    Full Text Available Background: Determination of the division history of T cells in vitro is helpful in the study of effector mechanisms against infections. Technique described here uses the intracellular fluorescent label carboxyfluorescein diacetate succinimidyl ester (CFSE to monitor the proliferation. Methods: In a cross sectional study, blood samples were collected from 7 volunteers with history of cutaneous leishmania­sis (CL and one healthy control from endemic areas in Isfahan province who referred to the Center for Research and Training in Skin Diseases and Leprosy (CRTSDL, then CD4+/CD8+ lymphocytes and CD14+ monocytes were isolated from peri­pheral blood mononuclear cells (PBMC using mAbs and magnetic nanoparticles. CFSE labeled CD4+ or CD8+ lympho­cytes cultured with autologous monocytes in the presence of PHA, SLA, live Leishmania major or as control with­out sti­mulation. Cells were harvested after 7 days and were analyzed using flow cytometry. Results: Five consecutive divisions were monitored separately. Stimulation of CD4+ or CD8+ lymphocytes from CL sub­jects with SLA showed a significant difference in proliferation comparing with unstimulated cells (P< 0.05. The signifi­cant difference in the percentages of CD4+ cells stimulated with SLA was revealed at different divisions for each subject. In CD8+ lymphocyte, significant stronger stimulation of SLA was evident later in the proliferation process. The mean number of divisions in both CD4+/CD8+ lymphocytes stimulated with SLA was significantly greater than when stimulated with live L. major (P=0.007 / P=0.012, respectively Conclusion: The percentage of divided cells might be calculated separately in each division. The cells remained active following CFSE staining and there is possibility of functional analysis simultaneously.

  17. Dysregulated angiogenesis in B-chronic lymphocytic leukemia: Morphologic, immunohistochemical, and flow cytometric evidence

    Directory of Open Access Journals (Sweden)

    Crawford Susan E

    2008-04-01

    Full Text Available Abstract Background The extent of enhanced bone marrow angiogenesis in chronic lymphocytic leukemia (CLL and relationship to proangiogenic factors and prognostic indicators is largely unexplored. Methods To further investigate the role of angiogenesis in CLL by evaluating the topography and extent of angiogenesis in a group of CLL bone marrow biopsies, to study the expression of pro and antiangiogenic vascular factors in CLL cells to more precisely document the cell types producing these factors, and to evaluate the role, if any, of localized hypoxia in upregulation of angiogenesis in CLL We used immunohistochemistry (IHC (n = 21 pts with antibodies to CD3 and CD20, proangiogenic (VEGF, HIF-1a and antiangiogenic (TSP-1 factors, and VEGF receptors -1 and -2 to examine pattern/extent of CLL marrow involvement, microvessel density (MVD, and angiogenic characteristics; flow cytometry (FC was performed on 21 additional cases for VEGF and TSP-1. Results CLL patients had higher MVD (23.8 vs 14.6, p~0.0002 compared to controls (n = 10. MVD was highest at the periphery of focal infiltrates, was not enhanced in proliferation centers, and was increased irrespective of the presence or absence of cytogenetic/immunophenotypic markers of aggressivity. By IHC, CLL cells were VEGF(+, HIF-1a (+, TSP-1(-, VEGFR-1(+, and VEGFR-2(+. By FC, CLL cells were 1.4–2.0-fold brighter for VEGF than T cells and were TSP-1(-. Conclusion CLL demonstrates enhanced angiogenesis, with increased MVD, upregulated VEGF and downregulated TSP-1. Upregulation of HIF-1a in all CLL cases suggests localized tissue hypoxia as an important stimulant of microvessel proliferation. The presence of VEGF receptors on CLL cells implies an autocrine effect for VEGF. Differences in MVD did not correlate with traditional genetic/immunophenotypic markers of aggressiveness.

  18. Flow cytometric bacterial cell counts challenge conventional heterotrophic plate counts for routine microbiological drinking water monitoring

    KAUST Repository

    Van Nevel, S.

    2017-02-08

    Drinking water utilities and researchers continue to rely on the century-old heterotrophic plate counts (HPC) method for routine assessment of general microbiological water quality. Bacterial cell counting with flow cytometry (FCM) is one of a number of alternative methods that challenge this status quo and provide an opportunity for improved water quality monitoring. After more than a decade of application in drinking water research, FCM methodology is optimised and established for routine application, supported by a considerable amount of data from multiple full-scale studies. Bacterial cell concentrations obtained by FCM enable quantification of the entire bacterial community instead of the minute fraction of cultivable bacteria detected with HPC (typically < 1% of all bacteria). FCM measurements are reproducible with relative standard deviations below 3% and can be available within 15 min of samples arriving in the laboratory. High throughput sample processing and complete automation are feasible and FCM analysis is arguably less expensive than HPC when measuring more than 15 water samples per day, depending on the laboratory and selected staining procedure(s). Moreover, many studies have shown FCM total (TCC) and intact (ICC) cell concentrations to be reliable and robust process variables, responsive to changes in the bacterial abundance and relevant for characterising and monitoring drinking water treatment and distribution systems. The purpose of this critical review is to initiate a constructive discussion on whether FCM could replace HPC in routine water quality monitoring. We argue that FCM provides a faster, more descriptive and more representative quantification of bacterial abundance in drinking water.

  19. Color encoded microbeads-based flow cytometric immunoassay for polycyclic aromatic hydrocarbons in food

    International Nuclear Information System (INIS)

    Meimaridou, Anastasia; Haasnoot, Willem; Noteboom, Linda; Mintzas, Dimitrios; Pulkrabova, Jana; Hajslova, Jana; Nielen, Michel W.F.

    2010-01-01

    Food contamination caused by chemical hazards such as persistent organic pollutants (POPs) is a worldwide public health concern and requires continuous monitoring. The chromatography-based analysis methods for POPs are accurate and quite sensitive but they are time-consuming, laborious and expensive. Thus, there is a need for validated simplified screening tools, which are inexpensive, rapid, have automation potential and can detect multiple POPs simultaneously. In this study we developed a flow cytometry-based immunoassay (FCIA) using a color-encoded microbeads technology to detect benzo[a]pyrene (BaP) and other polycyclic aromatic hydrocarbons (PAHs) in buffer and food extracts as a starting point for the future development of rapid multiplex assays including other POPs in food, such as polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs). A highly sensitive assay for BaP was obtained with an IC 50 of 0.3 μg L -1 using a monoclonal antibody (Mab22F12) against BaP, similar to the IC 50 of a previously described enzyme-linked immunosorbent assay (ELISA) using the same Mab. Moreover, the FCIA was 8 times more sensitive for BaP compared to a surface plasmon resonance (SPR)-based biosensor immunoassay (BIA) using the same reagents. The selectivity of the FCIAs was tested, with two Mabs against BaP for 25 other PAHs, including two hydroxyl PAH metabolites. Apart from BaP, the FCIAs can detect PAHs such as indenol[1,2,3-cd]pyrene (IP), benz[a]anthracene (BaA), and chrysene (CHR) which are also appointed by the European Food Safety Authority (EFSA) as suitable indicators of PAH contamination in food. The FCIAs results were in agreement with those obtained with gas chromatography-mass spectrometry (GC-MS) for the detection of PAHs in real food samples of smoked carp and wheat flour and has great potential for the future routine application of this assay in a simplex or multiplex format in combination with simplified extraction procedure which are

  20. Flow cytometric and microscopic analysis of the effect of tannic acid on plant nuclei and estimation of DNA content

    Czech Academy of Sciences Publication Activity Database

    Loureiro, J.; Rodriguez, E.; Doležel, Jaroslav; Santos, C.

    2006-01-01

    Roč. 98, - (2006), s. 515-527 ISSN 0305-7364 R&D Projects: GA MŠk(CZ) LC06004 Institutional research plan: CEZ:AV0Z50380511 Keywords : genome size * flow cytometry * nuclear DNA content Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.448, year: 2006

  1. Development of gammabeta thymocyte subsets during prenatal and postnatal ontogeny: a flow cytometric study in miniature pigs

    Czech Academy of Sciences Publication Activity Database

    Šinkora, Marek; Šinkorová, J.

    2003-01-01

    Roč. 87, č. 1 (2003), s. 101 ISSN 0165-2478 R&D Projects: GA ČR GA524/01/0919 Institutional research plan: CEZ:AV0Z5020903 Keywords : thymocyte subsets * cytometric Subject RIV: EE - Microbiology, Virology Impact factor: 1.710, year: 2003

  2. Flow Cytometric Quantification of Peripheral Blood Cell β-Adrenergic Receptor Density and Urinary Endothelial Cell-Derived Microparticles in Pulmonary Arterial Hypertension.

    Directory of Open Access Journals (Sweden)

    Jonathan A Rose

    Full Text Available Pulmonary arterial hypertension (PAH is a heterogeneous disease characterized by severe angiogenic remodeling of the pulmonary artery wall and right ventricular hypertrophy. Thus, there is an increasing need for novel biomarkers to dissect disease heterogeneity, and predict treatment response. Although β-adrenergic receptor (βAR dysfunction is well documented in left heart disease while endothelial cell-derived microparticles (Ec-MPs are established biomarkers of angiogenic remodeling, methods for easy large clinical cohort analysis of these biomarkers are currently absent. Here we describe flow cytometric methods for quantification of βAR density on circulating white blood cells (WBC and Ec-MPs in urine samples that can be used as potential biomarkers of right heart failure in PAH. Biotinylated β-blocker alprenolol was synthesized and validated as a βAR specific probe that was combined with immunophenotyping to quantify βAR density in circulating WBC subsets. Ec-MPs obtained from urine samples were stained for annexin-V and CD144, and analyzed by a micro flow cytometer. Flow cytometric detection of alprenolol showed that βAR density was decreased in most WBC subsets in PAH samples compared to healthy controls. Ec-MPs in urine was increased in PAH compared to controls. Furthermore, there was a direct correlation between Ec-MPs and Tricuspid annular plane systolic excursion (TAPSE in PAH patients. Therefore, flow cytometric quantification of peripheral blood cell βAR density and urinary Ec-MPs may be useful as potential biomarkers of right ventricular function in PAH.

  3. Flow cytometric cell sorting and in vitro pre-osteoinduction are not requirements for in vivo bone formation by human adipose-derived stromal cells.

    Science.gov (United States)

    Liu, Yunsong; Zhao, Yan; Zhang, Xiao; Chen, Tong; Zhao, Xianghui; Ma, Gui-e; Zhou, Yongsheng

    2013-01-01

    Human adipose-derived stromal cells (hASCs) are a promising cell source for bone tissue engineering. However, before the clinical application of hASCs for the treatment of bone defects, key questions require answers, including whether pre-osteoinduction (OI) and flow cytometric cell purification are indispensible steps for in vivo bone formation by hASCs. In this study, hASCs were purified by flow cytometric cell sorting (FCCS). The osteogenic capabilities of hASCs and purified hASCs with or without pre-osteoinduction were examined through in vitro and in vivo experiments. We found that pre-OI enhanced the in vitro osteogenic capacity of hASCs. However, 8 weeks after in vivo implantation, there were no significant differences between hASCs and hASCs that had undergone OI (hASCs+OI) or between purified hASCs and purified hASCs+OI (P>0.05). Interestingly, we also found that purified hASCs had an osteogenic potential similar to that of unpurified hASCs in vitro and in vivo. These results suggest that FCCS and in vitro pre-OI are not requirements for in vivo bone formation by hASCs.

  4. Flow cytometric cell sorting and in vitro pre-osteoinduction are not requirements for in vivo bone formation by human adipose-derived stromal cells.

    Directory of Open Access Journals (Sweden)

    Yunsong Liu

    Full Text Available Human adipose-derived stromal cells (hASCs are a promising cell source for bone tissue engineering. However, before the clinical application of hASCs for the treatment of bone defects, key questions require answers, including whether pre-osteoinduction (OI and flow cytometric cell purification are indispensible steps for in vivo bone formation by hASCs. In this study, hASCs were purified by flow cytometric cell sorting (FCCS. The osteogenic capabilities of hASCs and purified hASCs with or without pre-osteoinduction were examined through in vitro and in vivo experiments. We found that pre-OI enhanced the in vitro osteogenic capacity of hASCs. However, 8 weeks after in vivo implantation, there were no significant differences between hASCs and hASCs that had undergone OI (hASCs+OI or between purified hASCs and purified hASCs+OI (P>0.05. Interestingly, we also found that purified hASCs had an osteogenic potential similar to that of unpurified hASCs in vitro and in vivo. These results suggest that FCCS and in vitro pre-OI are not requirements for in vivo bone formation by hASCs.

  5. The evaluation of histo-blood group ABO typing by flow cytometric and PCR-amplification of specific alleles analyses and their application in clinical laboratories.

    Science.gov (United States)

    Aki, Kensaku; Izumi, Azusa; Hosoi, Eiji

    2012-01-01

    ABO antigens are oligosaccharide antigens, and are widely distributed on red blood and tissue cells as well as in saliva and body fluid. Therefore, these antigens are important not only for blood transfusion, but also for tissue cell and organ transplantations. Also, blood, hair, and seminal fluid are important sources of evidence at crime scenes, and these antigens are some of the most important markers for personal identification in forensic investigations. Here, we describe the development and use of quantitative analysis of A, B, and H antigens on red blood cells by employing flow cytometric analysis and the ABO genotyping method based on PCR-amplification of specific alleles (PASA) within DNA, especially from blood and saliva. In this study, flow cytometric analysis could be used to compare the differences between the expression of A and/or B and H antigens on red blood cells with various phenotypes, and the PASA method was able to determine the genotype of the type cisA(2)B(3) pedigree using only DNA extracted from saliva. These analysis methods are simple and useful for judging the ABO blood group system and genotyping, and are used widely throughout research and clinical laboratories and forensic fields.

  6. Development of a flow cytometric bead immunoassay and its assessment as a possible aid to potency evaluation of enterotoxaemia vaccines

    Directory of Open Access Journals (Sweden)

    Angela Buys

    2014-03-01

    Full Text Available Enterotoxaemia, an economically important disease of sheep, goats and calves, is caused by systemic effects of the epsilon toxin produced by the anaerobic bacterium Clostridium perfringens type D. The only practical means of controlling the occurrence of enterotoxaemia is to immunise animals by vaccination. The vaccine is prepared by deriving a toxoid from the bacterial culture filtrate and the potency of the vaccine is tested with the in vivo mouse neutralisation test (MNT. Due to ethical, economic and technical reasons, alternative in vitro assays are needed. In this study an indirect cytometric bead immunoassay (I-CBA was developed for use in vaccine potency testing and the results were compared with those obtained using an indirect enzyme-linked immunosorbent assay (I-ELISA and the MNT. Sera were collected from guinea pigs immunised with three different production batches of enterotoxaemia vaccine and the levels of anti-epsilon toxin antibodies were determined. Although the intra- and inter-assay variability was satisfactory, epsilon antitoxin levels determined by both the I-ELISA and indirect cytometric bead immunoassay (I-CBA tests were higher than those of the MNT assay. In contrast to the MNT, all of the serum samples were identified as having antitoxin levels above the required minimum (not less than 5 U/mL. These results indicate that the respective in vitro tests in their current formats are not yet suitable alternatives to the in vivo MNT. The growing demand for a more humane, cost-effective and efficient method for testing the potency of enterotoxaemia vaccines, however, provides a strong impetus for further optimisation and standardisation of the I-CBA assay but further analytical research is required.

  7. Effect of 17 beta-oestradiol on growth curves and flow cytometric DNA distribution of two human breast carcinomas grown in nude mice

    DEFF Research Database (Denmark)

    Brünner, N; Spang-Thomsen, M; Vindeløv, L

    1983-01-01

    The effect of 17 beta-oestradiol on a "receptor positive" and on a "receptor negative" human breast carcinoma grown in nude mice was studied. Experimental growth data were used to determine the effect on tumour growth. Flow cytometric DNA analysis (FCM) performed on tumour tissue obtained...... by sequential fine-needle aspirations was used to estimate the effect on the cell cycle. In the receptor-positive breast carcinoma, oestradiol induced complete tumour regression and characteristic cell cycle changes. In the receptor-negative breast carcinoma, no changes in tumour growth and cell cycle...... prior to any reduction in the tumour size, the results suggest that FCM may prove a valuable method in the early detection of tumour response to hormone treatment in human breast cancer....

  8. Japanese Society for Laboratory Hematology flow cytometric reference method of determining the differential leukocyte count: external quality assurance using fresh blood samples.

    Science.gov (United States)

    Kawai, Y; Nagai, Y; Ogawa, E; Kondo, H

    2017-04-01

    To provide target values for the manufacturers' survey of the Japanese Society for Laboratory Hematology (JSLH), accurate standard data from healthy volunteers were needed for the five-part differential leukocyte count. To obtain such data, JSLH required an antibody panel that achieved high specificity (particularly for mononuclear cells) using simple gating procedures. We developed a flow cytometric method for determining the differential leukocyte count (JSLH-Diff) and validated it by comparison with the flow cytometric differential leukocyte count of the International Council for Standardization in Haematology (ICSH-Diff) and the manual differential count obtained by microscopy (Manual-Diff). First, the reference laboratory performed an imprecision study of JSLH-Diff and ICSH-Diff, as well as performing comparison among JSLH-Diff, Manual-Diff, and ICSH-Diff. Then two reference laboratories and seven participating laboratories performed imprecision and accuracy studies of JSLH-Diff, Manual-Diff, and ICSH-Diff. Simultaneously, six manufacturers' laboratories provided their own representative values by using automated hematology analyzers. The precision of both JSLH-Diff and ICSH-Diff methods was adequate. Comparison by the reference laboratory showed that all correlation coefficients, slopes and intercepts obtained by the JSLH-Diff, ICSH-Diff, and Manual-Diff methods conformed to the criteria. When the imprecision and accuracy of JSLH-Diff were assessed at seven laboratories, the CV% for lymphocytes, neutrophils, monocytes, eosinophils, and basophils was 0.5~0.9%, 0.3~0.7%, 1.7~2.6%, 3.0~7.9%, and 3.8~10.4%, respectively. More than 99% of CD45 positive leukocytes were identified as normal leukocytes by JSLH-Diff. When JSLH-Diff method were validated by comparison with Manual-Diff and ICSH-Diff, JSLH-Diff showed good performance as a reference method. © 2016 John Wiley & Sons Ltd.

  9. Flow cytometric method for enumeration and characterization of newly released polymorphonuclear leukocytes from the bone marrow using 5'-bromo-2'-deoxyuridine.

    Science.gov (United States)

    Shih, Chih-Horng; Whalen, Beth A; Goto, Yukinobu; Hogg, James C; van Eeden, Stephan F

    2005-09-01

    Inflammation accelerates polymorphonuclear leukocyte (PMN) release from the bone marrow, and these PMNs are implicated in inappropriate tissue injury. We have previously developed a method using 5'-bromo-2'-deoxyuridine (BrdU) to study PMN kinetics using an immunocytochemical grading system of PMN on cytospin slides. The aim of this study was to develop a flow cytometric method to quantify the number of positively stained PMN and grade the intensity of staining for the transit time calculation of PMN through the marrow. Dividing myeloid progenitors in the marrow of rabbits were labeled with a pulse dosage of intravenous BrdU. BrdU-labeled PMN (PMN(BrdU)) were detected in the circulation using a FITC-conjugated anti-BrdU monoclonal antibody. The PMN(BrdU) were assigned to five groups according to their FITC intensity, and the transit times of PMN at different stages of development in the marrow were calculated. Results were compared using parallel immunocytochemical analysis of the same samples. In control animals, PMN(BrdU) in the circulation peaked at 72 h after BrdU labeling with 36.0% of PMN labeled. In normal rabbits, the transit times of PMN through the mitotic pool (49.5 +/- 4.2 h) and maturation pool (65.5 +/- 3.1 h) correlated well with immunocytochemical analysis and previously published values. Using this method, we demonstrated that exposure to air pollution particles accelerates the release of PMN(BrdU) from the marrow. We conclude that a flow cytometric approach for identifying BrdU-labeled leukocytes provides an objective and accurate method for studying leukocyte kinetics and behavior.

  10. Fluorescence in situ hybridization and sequential catalysed reporter deposition (2C-FISH for the flow cytometric sorting of freshwater ultramicrobacteria

    Directory of Open Access Journals (Sweden)

    Stefan M Neuenschwander

    2015-03-01

    Full Text Available Flow cytometric sorting is a powerful tool to physically separate cells within mixed microbial communities. If combined with phylogenetic staining (fluorescence in situ hybridization, FISH it allows to specifically sort defined genotypic microbial populations from complex natural samples. However, the targeted enrichment of freshwater ultramicrobacteria, such as members of the LD12 clade of Alphaproteobacteria (SAR11-IIIb, is still challenging. Current FISH protocols, even in combination with signal amplification by catalysed reporter deposition (CARD, are not sufficiently sensitive for the distinction of these bacteria from background noise by flow cytometry, presumably due to their low ribosome content and small cell sizes. We, therefore, modified a CARD based flow sorting protocol with the aim of increasing its sensitivity to a level sufficient for ultramicrobacteria. This was achieved by a second signal amplification step mediated by horseradish peroxidase labelled antibodies targeted to the fluorophores that were previously deposited by CARD-FISH staining. The protocol was tested on samples from an oligo-mesotrophic lake. Ultramicrobacteria affiliated with LD12 Alphaproteobacteria could be successfully sorted to high purity by flow cytometry. The ratios of median fluorescence signal to background ranged around 20, and hybridization rates determined by flow cytometry were comparable to those obtained by fluorescence microscopy. Potential downstream applications of our modified cell staining approach range from the analysis of microdiversity within 16S rRNA-defined populations to that of functional properties, such as the taxon-specific incorporation rates of organic substrates.

  11. Nuclear reactor core flow baffling

    International Nuclear Information System (INIS)

    Berringer, R.T.

    1979-01-01

    A flow baffling arrangement is disclosed for the core of a nuclear reactor. A plurality of core formers are aligned with the grids of the core fuel assemblies such that the high pressure drop areas in the core are at the same elevations as the high pressure drop areas about the core periphery. The arrangement minimizes core bypass flow, maintains cooling of the structure surrounding the core, and allows the utilization of alternative beneficial components such as neutron reflectors positioned near the core

  12. Effect of acidic pH on flow cytometric detection of bacteria stained with SYBR Green I and their distinction from background

    International Nuclear Information System (INIS)

    Baldock, Daniel; Nocker, Andreas; Nebe-von-Caron, Gerhard; Bongaerts, Roy

    2013-01-01

    Unspecific background caused by biotic or abiotic particles, cellular debris, or autofluorescence is a well-known interfering parameter when applying flow cytometry to the detection of microorganisms in combination with fluorescent dyes. We present here an attempt to suppress the background signal intensity and thus to improve the detection of microorganisms using the nucleic acid stain SYBR ® Green I. It has been observed that the fluorescent signals from SYBR Green I are greatly reduced at acidic pH. When lowering the pH of pre-stained samples directly prior to flow cytometric analysis, we hypothesized that the signals from particles and cells with membrane damage might therefore be reduced. Signals from intact cells, temporarily maintaining a neutral cytosolic pH, should not be affected. We show here that this principle holds true for lowering background interference, whereas the signals of membrane-compromised dead cells are only affected weakly. Signals from intact live cells at low pH were mostly comparable to signals without acidification. Although this study was solely performed with SYBR ® Green I, the principle of low pH flow cytometry (low pH-FCM) might hold promise when analyzing complex matrices with an abundance of non-cellular matter, especially when expanded to non-DNA binding dyes with a stronger pH dependence of fluorescence than SYBR Green I and a higher pK a value. (paper)

  13. Flow cytometric analysis of p21 protein expression on irradiated human lymphocytes; Analise por citometria de fluxo da expressao da proteina p21 em linfocitos humanos irradiados

    Energy Technology Data Exchange (ETDEWEB)

    Santos, N.F.G.; Amaral, A., E-mail: neyliane@gmail.com [Universidade Federal de Pernambuco (UFPE), Recife, PE (Brazil). Departamento de Energia Nuclear. Laboratorio de Modelagem e Biodosimetria Aplicada; Freitas-Silva, R. [Universidade Federal de Pernambuco (UFPE), Garanhuns, PE (Brazil). Departamento de Ciencias Naturais e Exatas; Pereira, V.R.A. [Fundacao Oswaldo Cruz (FIOCRUZ), Recife, PE (Brazil). Centro de Pesquisas Aggeu Magalhaes. Departamento de Imunologia. Lab. de Imunoparasitologia; Tasat, D.R. [Universidad Nacional de General San Martin, Buenos Aires (Argentina). Escuela de Ciencia y Tecnologia. Laboratorio de Biologia Celular del Pulmon

    2013-08-15

    Cell cycle blockage in G1 is a mechanism p21 protein-regulated and coupled to DNA damage response to permit genetic content analysis, damage repair and cell death. Analysis of proteins that participates of this response has progressed with new analytic tools, and data contributes to comprehension of radioinduced molecular events as well as to new approaches on practices that employ ionizing radiation. On this perspective, the aim of this research was to evaluate, by flow cytometry, p21 expression on irradiated human lymphocytes, maintained under different experimental conditions. Peripheral blood samples from 10 healthy subjects were irradiated with doses of 0 (non-irradiated), 1, 2 and 4 Gy. Lymphocytes were processed to analysis on ex vivo (no cultured) condition and after 24; 48 and 72 hours culture, with and without phytohemagglutinin stimulation. p21 protein expression levels were measured by flow cytometry, as percentage values. Results indicate that flow cytometric assay allows detection of changes on p21 expression, since it was detected significant increase on phytohemagglutinin-stimulated samples, for all times, against basal expression (ex vivo). However, it was not observed significant alterations on p21 protein radioinduced levels, for all doses, times and culture conditions analyzed. These results not indicate so p21 protein as bioindicator of ionizing radiation exposure. Nevertheless, data confirmation may to require analysis of a more numerous population. (author)

  14. Stereologic, histopathologic, flow cytometric, and clinical parameters in the prognostic evaluation of 74 patients with intraoral squamous cell carcinomas

    DEFF Research Database (Denmark)

    Bundgaard, T; Sørensen, Flemming Brandt; Gaihede, M

    1992-01-01

    , tumor size, and the TNM classification. RESULTS: The investigation showed a significant difference between the volume-weighted mean nuclear volume (nuclear vv) of oral leukoplakia (n = 29) and oral squamous cell carcinomas (P = 0.001). The value of the parameters as prognostic indicators of survival...

  15. Glycerol-treated nuclear suspensions - an efficient preservation method for flow cytometric analysis of plant samples

    Czech Academy of Sciences Publication Activity Database

    Kolář, Filip; Lučanová, Magdalena; Těšitel, J.; Loureiro, J.; Suda, Jan

    2012-01-01

    Roč. 20, č. 2 (2012), s. 303-315 ISSN 0967-3849 R&D Projects: GA ČR GD206/08/H049 Institutional research plan: CEZ:AV0Z60050516 Institutional support: RVO:67985939 Keywords : cytometry * ploidy * genome size Subject RIV: EF - Botanics Impact factor: 2.847, year: 2012

  16. Simple flow cytometric protocol of CD4+/CD8+ lymphocyte ratio assessment in bronchoalveolar lavage fluids from patients with interstitial lung diseases.

    Science.gov (United States)

    Szpechcinski, Adam; Kopinski, Piotr; Giedronowicz, Dorota; Rozy, Adriana; Jagus, Paulina; Szolkowska, Malgorzata; Chorostowska-Wynimko, Joanna

    2011-10-01

    To validate the fast and accurate flow cytometric (FCM) protocol using blood-standardized antibodies for alveolar lymphocyte subtyping with respect to standard immunocytochemistry (IC). FCM and IC were applied to immunophenotype T cell subsets in bronchoalveolar lavage (BAL) fluids from patients with interstitial lung diseases. Diagnostic BAL specimens from 50 patients with suspected sarcoidosis, idiopathic pulmonary fibrosis, and hypersensitivity pneumonitis were evaluated by both IC and FCM. In FCM, CD4+ and CD8+ T cells were identified by light scatter gating with CD3 selection using basic tricolor cytometer. Relative amounts of CD4+, CD8+ T cells, and CD4+/CD8+ ratios demonstrated by the FCM showed excellent, significant correlations with IC results. FCM values did not differ significantly from IC results. However, the sensitivity and specificity of conventional IC staining were not sufficient to assess CD4+/ CD8+ ratio in most idiopathic pulmonary fibrosis cases. Additionally, performing IC immunophenotyping in BAL samples with low lymphocyte content introduced a remarkable error into CD4+/CD8+ ratio assessment. FCM allowed reliable, precise, and fast T-cell subset measurement in all BAL samples, overcoming the IC disadvantages. Our validated FCM protocol provides diagnostically relevant CD4+/CD8+ ratio determination by simple light scatter gating strategy with CD3 selection.

  17. Molecular pathways of early CD105-positive erythroid cells as compared with CD34-positive common precursor cells by flow cytometric cell-sorting and gene expression profiling

    International Nuclear Information System (INIS)

    Machherndl-Spandl, S; Suessner, S; Danzer, M; Proell, J; Gabriel, C; Lauf, J; Sylie, R; Klein, H-U; Béné, M C; Weltermann, A; Bettelheim, P

    2013-01-01

    Special attention has recently been drawn to the molecular network of different genes that are responsible for the development of erythroid cells. The aim of the present study was to establish in detail the immunophenotype of early erythroid cells and to compare the gene expression profile of freshly isolated early erythroid precursors with that of the CD34-positive (CD34 + ) compartment. Multiparameter flow cytometric analyses of human bone marrow mononuclear cell fractions (n=20) defined three distinct early erythroid stages. The gene expression profile of sorted early erythroid cells was analyzed by Affymetrix array technology. For 4524 genes, a differential regulation was found in CD105-positive erythroid cells as compared with the CD34 + progenitor compartment (2362 upregulated genes). A highly significant difference was observed in the expression level of genes involved in transcription, heme synthesis, iron and mitochondrial metabolism and transforming growth factor-β signaling. A comparison with recently published data showed over 1000 genes that as yet have not been reported to be upregulated in the early erythroid lineage. The gene expression level within distinct pathways could be illustrated directly by applying the Ingenuity software program. The results of gene expression analyses can be seen at the Gene Expression Omnibus repository

  18. Commercially Available Antibodies to Human Tumour Necrosis Factor-α Tested for Cross-Reactivity with Ovine and Bovine Tumour Necrosis Factor-α using Flow Cytometric Assays

    Directory of Open Access Journals (Sweden)

    Waller K Persson

    2004-06-01

    Full Text Available A thorough understanding of the immune system, including the role of different cytokines, during inflammatory diseases in ruminants could lead to the development of new diagnostic methods and treatments. Tumour necrosis factor-α (TNF-α is an important cytokine in the onset of the inflammatory responses. Unfortunately, the number of studies on cytokines, like TNF-α, in ruminants is limited due to a lack of species-specific reagents. As cytokines have remained rather conserved during evolution, cross-reactivity between animal species may occur. Therefore, the aim of the present study was to investigate 5 commercially available antibodies against human TNF-α for their ability to cross-react with ovine and/or bovine TNF-α, using a bead-based flow cytometric method. Two of the antibody clones (Mab 11 and 6401.1111 showed cross reactivity with ovine recombinant TNF-α in concentrations above 2.5 ng/ml. However, none of the antibodies detected TNF-α in bovine milk, or serum containing known concentrations of bovine TNF-α, as earlier determined with ELISA. The results could be due to inability of the antibodies to cross-react between species, but quenching of the signal by matrix proteins might also have lowered the response.

  19. Response of Syngonium podophyllum L. ‘White Butterfly’ shoot cultures to alternative media additives and gelling agents, and flow cytometric analysis of regenerants

    Directory of Open Access Journals (Sweden)

    JAIME A. TEIXEIRA DA SILVA

    2015-05-01

    Full Text Available Abstract. Teixeira da Silva JA. 2015. Response of Syngonium podophyllum L. ‘White Butterfly’ shoot cultures to alternative media additives and gelling agents, and flow cytometric analysis of regenerants. Nusantara Bioscience 7: 26-32. Syngonium podophyllum L. (arrowhead vine is a popular leafy indoor pot plant whose tissue culture has been established, primarily through in vitro shoot culture, but several interesting aspects have not yet been explored. In this study, cv. ‘White Butterfly’ was used to investigate the response of shoot formation to alternative gelling agents and media additives. Gellan gum (Gelrite® at 2 g/L resulted in greater leaf production, plantlet fresh weight and higher chlorophyll content (SPAD value than all other gelling agents tested, including agar, Bacto agar, phytagel, oatmeal agar, potato dextrose agar, barley starch and corn starch, when on a basal Hyponex® (NPK = 6.5: 6: 19; 3 g/L medium. Several alternative liquid medium additives tested (low and full fat milk, Coca-Cola®, coffee, Japanese green, Oolong and Darjeeling teas negatively impacted plant growth, stunted roots and decreased chlorophyll content (SPAD value of leaves. Plant growth on medium with refined sucrose or table sugar responded similarly. Poor growth was observed when crude extract from a high rebaudioside-containing stevia (Stevia rebaudiana Bertoni line - an artificial sweetener - was used. Leaf tissue from the control did not show any endopolyploidy but low levels of endopolyploidy (8C were detected in some treatments.

  20. Effect of chronic pesticide exposure on murine cornea: a histopathological, cytological and flow cytometric approach to study ocular damage by xenobiotics.

    Science.gov (United States)

    Sanyal, Shalini; Das, Prosun; Law, Sujata

    2016-02-01

    Pesticide exposure can occur directly or indirectly in an occupational setting or otherwise. The health hazards of pesticides have long been studied; however, little is known about the ocular insult of these potent chemicals. In this study, we examined the consequences of long-term pesticide exposure on the ocular tissue in animal model with special focus on the cornea. Swiss Albino mice were sacrificed to obtain the eye globes and various cytological, cytotoxic and histological evaluations, in vitro growth kinetic studies and flow cytometric analyses of select cytokeratins were performed to determine the structural and functional damage due to pesticide exposure. Our study revealed the detrimental impact of this xenobiotic insult by cataloguing the damage to each layer of the cornea wherein it was discovered that all the functional layers as well as the membranes were compromised. We hope that our investigation will pave the way for future studies in this oft overlooked area of affront caused by pesticide exposure to the ocular surface.

  1. Lymphocyte subset enumeration in HIV seronegative and HIV-1 seropositive adults in Dar es Salaam, Tanzania: determination of reference values in males and females and comparison of two flow cytometric methods.

    Science.gov (United States)

    Urassa, W K; Mbena, E M; Swai, A B; Gaines, H; Mhalu, F S; Biberfeld, G

    2003-06-01

    The level of CD4(+) T-lymphocytes represents a useful marker with which to monitor the progression of HIV infection. Sex and geographical differences in the reference values of lymphocyte subsets have been reported. We have compared two flow cytometric methods (MultiSET and SimulSET) for the quantification of lymphocyte subsets using whole blood from 92 HIV seropositive and 241 seronegative adults, and determined the reference values of lymphocyte subsets in HIV seronegative Tanzanian subjects. In seronegative Tanzanian subjects, the percentages of CD3(+) and CD4(+) T-lymphocytes and the CD4(+):CD8(+) T-lymphocyte ratios were lower while the percentage of natural killer cells was higher compared to the levels of the corresponding parameters reported for Europeans. Seronegative Tanzanian females had significantly higher levels of CD3(+) and CD4(+) T-lymphocytes and CD4(+):CD8(+) T-lymphocyte ratios compared to seronegative males. The correlation coefficients of CD3(+), CD4(+) and CD8(+) T lymphocyte counts and percentages obtained by the two flow cytometric methods were high. The median values of the number of CD4(+) T-lymphocytes obtained by the two methods were not significantly different. In conclusion, determination of the reference values of lymphocyte subsets in HIV seronegative Tanzanian adults showed significant sex differences and differences in percentage values compared to those reported in certain other geographical areas. There was acceptable agreement in the levels of CD4(+) T-lymphocyte values obtained by the two flow cytometric methods.

  2. Stereologic, histopathologic, flow cytometric, and clinical parameters in the prognostic evaluation of 74 patients with intraoral squamous cell carcinomas

    DEFF Research Database (Denmark)

    Bundgaard, T; Sørensen, Flemming Brandt; Gaihede, M

    1992-01-01

    BACKGROUND AND METHODS: A consecutive series of all 78 incident cases of intraoral squamous cell carcinoma occurring during a 2-year period in a population of 1.4 million inhabitants were evaluated by histologic score (the modified classification of Jacobsson et al.), flow cytometry, stereology, ...

  3. Development of a flow cytometric method to analyze subpopulations of bacteria in probiotic products and dairy starters

    NARCIS (Netherlands)

    Bunthof, C.J.; Abee, T.

    2002-01-01

    Flow cytometry (FCM) is a rapid and sensitive technique that can determine cell numbers and measure various physiological characteristics of individual cells by using appropriate fluorescent probes. Previously, we developed an FCM assay with the viability probes carboxyfluorescein diacetate (cFDA)

  4. [Standardization of the quantitative flow cytometric test with anti-D antibodies for fetomaternal hemorrhage in RhD negative women].

    Science.gov (United States)

    Spychalska, Justyna; Uhrynowska, Małgorzata; Pyl, Hanna; Klimczak-Jajor, Edyta; Kopeć, Izabella; Peciakowska, Małgorzata; Gutowska, Renata; Gawlak, Maciej; Słomska, Sylwia; Dąbkowska, Syiwia; Szczecina, Roman; Dębska, Marzena; Brojer, Ewa

    2015-07-01

    In order to determine the appropriate dose of anti-D immunoglobulin to be administered as a preventive measure against hemolytic disease of the fetus/newborn in the subsequent pregnancy it is necessary to assess the number of fetal red blood cells that infiltrate/penetrate into the maternal circulation as a result of fetomaternal hemorrhage (FMH). One of the quantitative methods of FMH analysis is based on flow cytometry (FACS) which makes use of monoclonal antibodies to RhD antigen (anti-D test). The aim of the study was to further develop the method, evaluate its sensitivity and reproducibility and to compare it with the test based on the detection of fetal hemoglobin (HbF). The FACS study involved 20 RhD negative pregnant women and 80 RhD negative women after delivery. The following monoclonal antibodies were used: BRAD 3 FITC (anti-RhD antigen), CD45 PerCP (anti leukocyte antigen CD45), and anti-HbF PE. The fluorescence intensity of cells incubated with BRAD 3 FITC was demonstrated to depend on the RhD antigen expression, though the anti-D test also detects the weak D variant. The CD45 PerCP antibodies increased the sensitivity of anti-D test since they eliminated the leukocytes which non-specifically bind anti-D from the analysis. The presence of anti-D antibodies in maternal plasma does not affect the quantitative assessment of the fetal RhD positive fetal cells with BRAD 3 FITC. In case of FMH, the results of the anti-D test were similar to those with anti-HbF antibodies. The flow cytometric test with anti-D and anti-CD45 is useful in the assessment of the fetomaternal hemorrhage in RhD negative women. The sensitivity of the test is estimated at 0.05%.

  5. Flow cytometric assessment of chicken T cell-mediated immune responses after Newcastle disease virus vaccination and challenge

    DEFF Research Database (Denmark)

    Dalgaard, T. S.; Norup, L. R.; Pedersen, A.R.

    2010-01-01

    . Despite a delayed NDV-specific antibody response to vaccination, L133 appeared to be better protected than L130 in the subsequent infection challenge as determined by the presence of viral genomes. Peripheral blood was analyzed by flow cytometry and responses in vaccinated/challenged birds were studied...... by 5-color immunophenotyping as well as by measuring the proliferative capacity of NDV-specific T cells after recall stimulation. Immunophenotyping identified L133 as having a significantly lower CD4/CD8 ratio and a lower frequency of γδ T cells than L130 in the peripheral T cell compartment...

  6. Flow cytometric sexing of spider sperm reveals an equal sperm production ratio in a female-biased species

    DEFF Research Database (Denmark)

    Vanthournout, Bram; Deswarte, K; Hammad, H

    2014-01-01

    that other factors influence sex ratio variation. In this paper, we investigate whether this additional variation can be explained by the unequal production of male- and female-determining sperm cells during sperm production. Using flow cytometry, we show that males produce equal amounts of male- and female......-determining sperm cells; thus bias in sperm production does not contribute to the sex ratio bias observed in this species. This demonstrates that other factors such as parental genes suppressing endosymbiont effects and cryptic female choice might play a role in sex allocation in this species....

  7. Improved flow cytometric identification of myelopoiesis by the simultaneous labelling with CD13, CD14 and CD66 monoclonal antibodies

    DEFF Research Database (Denmark)

    Bonde, J; Meyer, K; Broe, M K

    1996-01-01

    in the fast determination of remission state. In MDS, the immature myeloid component could be distinguished in patients defined according to the FAB classification with the possibility of identifying aberrant phenotypes, the assay should also be of interest in other myeloproliferative disorders. Moreover......The aim of the present study was to increase our knowledge of myelopoiesis evaluated by flow cytometry. We therefore designed a triple-marker assay employing monoclonal antibodies against the CD13 (immature), the CD14 (monocytic), and the CD66 (mature myeloid) antigens using three...

  8. Enumeration of extracellular vesicles by a new improved flow cytometric method is comparable to fluorescence mode nanoparticle tracking analysis.

    Science.gov (United States)

    Pasalic, Leonardo; Williams, Rebekka; Siupa, Agnieszka; Campbell, Heather; Henderson, Michelle J; Chen, Vivien M Y

    2016-05-01

    Extracellular vesicles (EVs) play a role in a variety of physiological and pathological processes. However, use of EVs as biomarkers has been hampered by limitations of current detection and enumeration methods. We compared fluorescence-threshold flow cytometry (FT-FC) to nanoparticle tracking analysis (NTA) for enumeration of cell culture-derived EVs. FT-FC and NTA utilising fluorescence mode (F-NTA) enumerated similar numbers of EVs stained with a membrane dye PKH67. Both methods were sufficiently sensitive to detect cell-derived EVs above the background of culture medium. Light scatter NTA (LS-NTA) detected 10-100× more particles than either fluorescence-based method but demonstrated poor specificity. F-NTA appeared to have better sensitivity for vesicles, however, the FT-FC method combined direct enumeration of EVs with high sensitivity and specificity in the >100nm range. Due to wider availability and higher degree of automation and standardisation, FT-FC is a reasonable surrogate to F-NTA for quantification of EVs. Extracellular vesicles are small particles, which can act as tools for intercellular communication. One recent area of interest in EVs is their potentials as biomarkers. In this article, the authors investigated and compared fluorescence-threshold flow cytometry (FT-FC) to nanoparticle tracking analysis (NTA) for the detection of EVs and showed that FT- FC method could be more advantageous. This technique should provide a new alternative for the future. Crown Copyright © 2016. Published by Elsevier Inc. All rights reserved.

  9. Flow cytometric minimal residual disease monitoring in children with acute lymphoblastic leukemia treated by regimens with reduced intensity

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    A. M. Popov

    2015-01-01

    Full Text Available 191 consecutive unselected children with acute lymphoblastic leukemia aged from 1 to 16 years were enrolled in the study. Bone marrow samples were obtained at the time of initial diagnostics as well as at days 15 (n = 188, 36 (n = 191, and 85 (n = 187 of remission induction. Minimal residual disease (MRD was assessed by 6–10-color flow cytometry. Flow cytometry data at day 15 allowed distinguishing three patients groups with significantly different outcome (p ˂ 0.0001: 35.64 % patients with MRD < 0.1 % represented 5-year event-free survival (EFS of 100 %; 48.40 % cases with 0.1 % ≤ MRD< 10 % had EFS 84.6 ± 4.2 %; 15.96 % patients with very high MRD (≥ 10 % belonged to group with poor outcome (EFS 56.7 ± 9.0 %. At the end of remission induction (day 36 36 children (18.85 % with MRD higher than 0.1 % had significantly worse outcome compared to remaining ones (EFS 49.4 ± 9.0 and 93.5 ± 2.1 % respectively; p ˂ 0.0001. From a clinical standpoint it is relevant to evaluate both low-risk and high-risk criteria. Multivariate analysis showed that day 15 MRD data is better for low-risk patients definition while end-induction MRD is the strongest unfavorable prognostic factor.

  10. Influence of a radioprotector WR-638 on the lymphoid compartment of the irradiated rat thymus: a flow cytometric analysis

    International Nuclear Information System (INIS)

    Dragojevic-Simic, V.; Colic, M.; Gasic, S.

    1994-01-01

    The T cell composition of the thymus of X-ray irradiated (3.5 Gy) Wistar rat protected with WR-638 was analyzed by flow cytometry using monoclonal antibodies directed to the Thy 1.1, CD43, CD2, CD5, CD4, CD8 and class I and II MHC antigens. It was shown that this dose of X-rays caused cyclic changes in thymic cellularity manifested as: primary involution (until day 2), primary regeneration (from days 2 to 14), secondary involution (from days 14 to 21) and secondary regeneration (from days 21 to 30). WR-638 reduced the magnitude of thymocyte depletion in the primary involutive phase of the irradiated thymi. (author)

  11. Genetic stock assessment of spawning arctic cisco (Coregonus autumnalis) populations by flow cytometric determination of DNA content.

    Science.gov (United States)

    Lockwood, S F; Bickham, J W

    1991-01-01

    Intraspecific variation in cellular DNA content was measured in five Coregonus autumnalis spawning populations from the Mackenzie River drainage, Canada, using flow cytometry. The rivers assayed were the Peel, Arctic Red, Mountain, Carcajou, and Liard rivers. DNA content was determined from whole blood preparations of fish from all rivers except the Carcajou, for which kidney tissue was used. DNA content measurements of kidney and blood preparations of the same fish from the Mountain River revealed statistically indistinguishable results. Mosaicism was found in blood preparations from the Peel, Arctic Red, Mountain, and Liard rivers, but was not observed in kidney tissue preparations from the Mountain or Carcajou rivers. The Liard River sample had significantly elevated mean DNA content relative to the other four samples; all other samples were statistically indistinguishable. Significant differences in mean DNA content among spawning stocks of a single species reinforces the need for adequate sample sizes of both individuals and populations when reporting "C" values for a particular species.

  12. Flow cytometric measurement of the metabolism of benzo[a]pyrene by mouse liver cells in culture

    International Nuclear Information System (INIS)

    Bartholomew, J.C.; Wade, C.G.; Dougherty, K.K.

    1984-01-01

    The metabolism of benzo[a]pyrene in individual cells was monitored by flow cytometry. The measurements are based on the alterations that occur in the fluorescence emission spectrum of benzo[a]pyrene when it is converted to various metabolites. Using present instrumentation the technique could easily detect 1x10 6 molecules per cells of benzo[a]pyrene and 1x10 7 molecules per cell of the diol epoxide. The analysis of C3H IOT 1/2 mouse fibroblasts growing in culture indicated that there was heterogeneity in the conversion of the parent compound into diol epoxide derivatives suggesting that some variation in sensitivity to transformation by benzo[a]pyrene may be due to differences in cellular metabolism. The technique allows sensitive detection of metabolites in viable cells, and provides a new approach to the study of factors that influence both metabolism and transformation. (orig.)

  13. Flow cytometric quantification of all phases of the cell cycle and apoptosis in a two-color fluorescence plot.

    Directory of Open Access Journals (Sweden)

    Christine Vignon

    Full Text Available An optimal technology for cell cycle analysis would allow the concomitant measurement of apoptosis, G0, G1, S, G2 and M phases in combination with cell surface phenotyping. We have developed an easy method in flow cytometry allowing this discrimination in an only two-color fluorescent plot. It is based on the concomitant use of 7-amino-actinomycin D and the antibodies anti-Ki67 and anti-phospho(Ser10-histone H3, both conjugated to Alexa Fluor®488 to discriminate G0 and M phases, respectively. The method is particularly valuable in a clinical setting as verified in our laboratory by analyzing human leukemic cells from marrow samples or after exposure to cell cycle modifiers.

  14. Evaluation of flow cytometric immunophenotyping and DNA analysis for detection of malignant cells in serosal cavity fluids.

    Science.gov (United States)

    Sayed, Douaa M; el-Attar, Madiha M; Hussein, Aliaa A R Mohamed

    2009-07-01

    The serosal cavities are frequent sites of tumor metastasis. The distinction between carcinoma cells, inflammatory cells, and reactive or malignant mesothelial cells can be difficult in cytology. Multicolor flow cytometry (FCM) provides the opportunity to evaluate multiple antigens simultaneously, making it possible to characterize various cell populations. In this study, we aimed to assess the diagnostic accuracy of FCM immunophenotyping and DNA in comparison with serum tumor markers and classic cytology for detection of malignant cells in pleural and ascitic fluids. One hundred and nineteen samples of body cavity fluids were analyzed. Immunophenotyping was performed by four-color immunofluorescent staining using monoclonal antibodies against Ber-EP4, cytokeratin, CD3, and CD45. The DNA analysis by FCM was also performed. In addition, serum CA19-9, CEA, AFP, and CA125 were analyzed. Ber-EP4 marker had the highest sensitivity (73%) and specificity (95.5%) in the detection of carcinoma cells in serous fluid and correlated with cytology in most of cases (73%). The mean of DI differed statistically in patients with malignant effusions than in benign one. DI showed no difference in fluids due to infiltration of malignant epithelial cells or hematopoietic malignancy or due to hepatocellular carcinoma developing in cirrhotic liver. Thus, flow cytometry appears to aid not only in the detection of malignant cells but also in the characterization of cell type. On the other hand, although DNA ploidy examination had better sensitivity; it had no advantage over conventional cytopathological examination in identification of malignant cells. 2009 Wiley-Liss, Inc.

  15. Optimized multiparametric flow cytometric analysis of circulating endothelial cells and their subpopulations in peripheral blood of patients with solid tumors: a technical analysis.

    Science.gov (United States)

    Zhou, Fangbin; Zhou, Yaying; Yang, Ming; Wen, Jinli; Dong, Jun; Tan, Wenyong

    2018-01-01

    Circulating endothelial cells (CECs) and their subpopulations could be potential novel biomarkers for various malignancies. However, reliable enumerable methods are warranted to further improve their clinical utility. This study aimed to optimize a flow cytometric method (FCM) assay for CECs and subpopulations in peripheral blood for patients with solid cancers. An FCM assay was used to detect and identify CECs. A panel of 60 blood samples, including 44 metastatic cancer patients and 16 healthy controls, were used in this study. Some key issues of CEC enumeration, including sample material and anticoagulant selection, optimal titration of antibodies, lysis/wash procedures of blood sample preparation, conditions of sample storage, sufficient cell events to enhance the signal, fluorescence-minus-one controls instead of isotype controls to reduce background noise, optimal selection of cell surface markers, and evaluating the reproducibility of our method, were integrated and investigated. Wilcoxon and Mann-Whitney U tests were used to determine statistically significant differences. In this validation study, we refined a five-color FCM method to detect CECs and their subpopulations in peripheral blood of patients with solid tumors. Several key technical issues regarding preanalytical elements, FCM data acquisition, and analysis were addressed. Furthermore, we clinically validated the utility of our method. The baseline levels of mature CECs, endothelial progenitor cells, and activated CECs were higher in cancer patients than healthy subjects ( P technical issues found in previously published assays and validated the reproducibility and sensitivity of our proposed method. Future work is required to explore the potential of our optimized method in clinical oncologic applications.

  16. Flow cytometric monitoring of bacterioplankton phenotypic diversity predicts high population-specific feeding rates by invasive dreissenid mussels.

    Science.gov (United States)

    Props, Ruben; Schmidt, Marian L; Heyse, Jasmine; Vanderploeg, Henry A; Boon, Nico; Denef, Vincent J

    2018-02-01

    Species invasion is an important disturbance to ecosystems worldwide, yet knowledge about the impacts of invasive species on bacterial communities remains sparse. Using a novel approach, we simultaneously detected phenotypic and derived taxonomic change in a natural bacterioplankton community when subjected to feeding pressure by quagga mussels, a widespread aquatic invasive species. We detected a significant decrease in diversity within 1 h of feeding and a total diversity loss of 11.6 ± 4.1% after 3 h. This loss of microbial diversity was caused by the selective removal of high nucleic acid populations (29 ± 5% after 3 h). We were able to track the community diversity at high temporal resolution by calculating phenotypic diversity estimates from flow cytometry (FCM) data of minute amounts of sample. Through parallel FCM and 16S rRNA gene amplicon sequencing analysis of environments spanning a broad diversity range, we showed that the two approaches resulted in highly correlated diversity measures and captured the same seasonal and lake-specific patterns in community composition. Based on our results, we predict that selective feeding by invasive dreissenid mussels directly impacts the microbial component of the carbon cycle, as it may drive bacterioplankton communities toward less diverse and potentially less productive states. © 2017 Society for Applied Microbiology and John Wiley & Sons Ltd.

  17. Transmission electron microscopic morphological study and flow cytometric viability assessment of Acinetobacter baumannii susceptible to Musca domestica cecropin.

    Science.gov (United States)

    Gui, Shuiqing; Li, Rongjiang; Feng, Yongwen; Wang, Sanming

    2014-01-01

    Multidrug-resistant (MDR) Acinetobacter baumannii infections are difficult to treat owing to the extremely limited armamentarium. Expectations about antimicrobial peptides' use as new powerful antibacterial agents have been raised on the basis of their unique mechanism of action. Musca domestica cecropin (Mdc), a novel antimicrobial peptide from the larvae of Housefly (Musca domestica), has potently active against Gram-positive and Gram-negative bacteria standard strain. Here we evaluated the antibacterial activity of Mdc against clinical isolates of MDR-A. baumannii and elucidate the related antibacterial mechanisms. The minimal inhibitory concentration (MIC) of Mdc was 4 μg/mL. Bactericidal kinetics of Mdc revealed rapid killing of A. baumannii (30 min). Flow cytometry using viability stain demonstrated that Mdc causes A. baumannii membrane permeabilization in a concentration- and time-dependent process, which correlates with the bactericidal action. Moreover, transmission electron microscopic (TEM) examination showed that Mdc is capable of disrupting the membrane of bacterial cells, resulting in efflux of essential cytoplasmic components. Overall, Mdc could be a promising antibacterial agent for MDR-A. baumannii infections.

  18. Transmission Electron Microscopic Morphological Study and Flow Cytometric Viability Assessment of Acinetobacter baumannii Susceptible to Musca domestica cecropin

    Directory of Open Access Journals (Sweden)

    Shuiqing Gui

    2014-01-01

    Full Text Available Multidrug-resistant (MDR Acinetobacter baumannii infections are difficult to treat owing to the extremely limited armamentarium. Expectations about antimicrobial peptides' use as new powerful antibacterial agents have been raised on the basis of their unique mechanism of action. Musca domestica cecropin (Mdc, a novel antimicrobial peptide from the larvae of Housefly (Musca domestica, has potently active against Gram-positive and Gram-negative bacteria standard strain. Here we evaluated the antibacterial activity of Mdc against clinical isolates of MDR-A. baumannii and elucidate the related antibacterial mechanisms. The minimal inhibitory concentration (MIC of Mdc was 4 μg/mL. Bactericidal kinetics of Mdc revealed rapid killing of A. baumannii (30 min. Flow cytometry using viability stain demonstrated that Mdc causes A. baumannii membrane permeabilization in a concentration- and time-dependent process, which correlates with the bactericidal action. Moreover, transmission electron microscopic (TEM examination showed that Mdc is capable of disrupting the membrane of bacterial cells, resulting in efflux of essential cytoplasmic components. Overall, Mdc could be a promising antibacterial agent for MDR-A. baumannii infections.

  19. The combination of kinetic and flow cytometric semen parameters as a tool to predict fertility in cryopreserved bull semen.

    Science.gov (United States)

    Gliozzi, T M; Turri, F; Manes, S; Cassinelli, C; Pizzi, F

    2017-11-01

    Within recent years, there has been growing interest in the prediction of bull fertility through in vitro assessment of semen quality. A model for fertility prediction based on early evaluation of semen quality parameters, to exclude sires with potentially low fertility from breeding programs, would therefore be useful. The aim of the present study was to identify the most suitable parameters that would provide reliable prediction of fertility. Frozen semen from 18 Italian Holstein-Friesian proven bulls was analyzed using computer-assisted semen analysis (CASA) (motility and kinetic parameters) and flow cytometry (FCM) (viability, acrosomal integrity, mitochondrial function, lipid peroxidation, plasma membrane stability and DNA integrity). Bulls were divided into two groups (low and high fertility) based on the estimated relative conception rate (ERCR). Significant differences were found between fertility groups for total motility, active cells, straightness, linearity, viability and percentage of DNA fragmented sperm. Correlations were observed between ERCR and some kinetic parameters, and membrane instability and some DNA integrity indicators. In order to define a model with high relation between semen quality parameters and ERCR, backward stepwise multiple regression analysis was applied. Thus, we obtained a prediction model that explained almost half (R 2=0.47, Pfertility. Once the accuracy of fertility prediction has been confirmed, the model developed in the present study could be used by artificial insemination centers for bull selection or for elimination of poor fertility ejaculates.

  20. Flow cytometric analysis of FSHR, BMRR1B, LHR and apoptosis in granulosa cells and ovulation rate in merino sheep.

    Science.gov (United States)

    Regan, Sheena L P; McFarlane, James R; O'Shea, Tim; Andronicos, Nicholas; Arfuso, Frank; Dharmarajan, Arun; Almahbobi, Ghanim

    2015-08-01

    The aim of the present study was to determine the direct cause of the mutation-induced, increased ovulation rate in Booroola Merino (BB) sheep. Granulosa cells were removed from antral follicles before ovulation and post-ovulation from BB (n=5) and WT (n=12) Merino ewes. Direct immunofluorescence measurement of mature cell surface receptors using flow cytometry demonstrated a significant up-regulation of FSH receptor (FSHR), transforming growth factor beta type 1, bone morphogenetic protein receptor (BMPR1B), and LH receptor (LHR) in BB sheep. The increased density of FSHR and LHR provide novel evidence of a mechanism for increasing the number of follicles that are recruited during dominant follicle selection. The compounding increase in receptors with increasing follicle size maintained the multiple follicles and reduced the apoptosis, which contributed to a high ovulation rate in BB sheep. In addition, we report a mutation-independent mechanism of down-regulation to reduce receptor density of the leading dominant follicle in sheep. The suppression of receptor density coincides with the cessation of mitogenic growth and steroidogenic differentiation as part of the luteinization of the follicle. The BB mutation-induced attenuation of BMPR1B signaling led to an increased density of the FSHR and LHR and a concurrent reduction in apoptosis to increase the ovulation rate. The role of BMPs in receptor modulation is implicated in the development of multiple ovulations. © 2015 Society for Reproduction and Fertility.

  1. Multiparameter flow cytometric analysis of CD4 and CD8 T cell subsets in young and old people

    Directory of Open Access Journals (Sweden)

    Özcelik Dennis

    2008-07-01

    Full Text Available Abstract Background T cell-mediated immunity in elderly people is compromised in ways reflected in the composition of the peripheral T cell pool. The advent of polychromatic flow cytometry has made analysis of cell subsets feasible in unprecedented detail. Results Here we document shifts in subset distribution within naïve (N, central memory (CM and effector memory (EM cells defined by CD45RA and CCR7 expression in the elderly, additionally using the costimulatory receptors CD27 and CD28, as well as the coinhibitory receptors CD57 and KLRG-1, to further dissect these. Although differences between young and old were more marked in CD8 than in CD4 cells, a similar overall pattern prevailed in both. Thus, the use of all these markers together, and inclusion of assays of proliferation and cytokine secretion, may enable the construction of a differentiation scheme applicable to CD4 as well as CD8 cells, with the model (based on Romero et al. suggesting the progression N→CM→EM1→EM2→pE1→pE2→EM4→EM3→E end-stage non-proliferative effector cells. Conclusion Overall, the results suggest that both differences in subset distribution and differences between subsets are responsible for age-related changes in CD8 cells but that differences within rather than between subsets are more prominent for CD4 cells.

  2. Flow cytometric investigations of diploid and tetraploid plants and in vitro cultures of Datura stramonium and Hyoscyamus niger.

    Science.gov (United States)

    Weber, Jost; Georgiev, Vasil; Pavlov, Atanas; Bley, Thomas

    2008-10-01

    Plant in vitro systems are valuable sources for the production of biological active substances. However, changed profiles of secondary metabolites, and low, variable yields possibly caused by genetic instabilities complicate their industrial implementation. DNA profiling of plant in vitro cultures may provide data for the selection of highly producing in vitro cultures. Diploid and tetraploid Datura stramonium and Hyoscyamus niger plant as well as calli, and hairy root lines derived from them were analyzed by flow cytometry. Plant in vitro cultures undergo several cycles of endoreduplication more than the explants from which they were obtained. The highest cycle values were observed in calli (e.g. 1.19 for diploid H. niger) possibly induced by the growth factors. However, hairy roots cultivated without growth factor exhibited significant degrees of endoreduplication (cycle value 0.88 for diploid H. niger). Sets of five hairy root lines from each plant and ploidy level showed consistent within-set ploidy patterns. The ploidy profiles of investigated plant in vitro and in vivo differ. For the first time we report that hairy roots of two Solanaceae species undergo endoreduplication. Theploidy profiles of in vitro cultures (hairy roots and calli) seem to be influenced by the genome size, the growth factors applied, and the type of in vitro culture. The transformation of several hairy root lines showed no differences in the ploidy patterns. Copyright 2008 International Society for Advancement of Cytometry.

  3. Flow Cytometric Clinical Immunomonitoring Using Peptide–MHC Class II Tetramers: Optimization of Methods and Protocol Development

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    Diahann T. S. L. Jansen

    2018-01-01

    Full Text Available With the advent of novel strategies to induce tolerance in autoimmune and autoimmune-like conditions, clinical trials of antigen-specific tolerizing immunotherapy have become a reality. Besides safety, it will be essential to gather mechanistic data on responding CD4+ T cells to assess the effects of various immunomodulatory approaches in early-phase trials. Peptide–MHC class II (pMHCII multimers are an ideal tool for monitoring antigen-specific CD4+ T cell responses in unmanipulated cells directly ex vivo. Various protocols have been published but there are reagent and assay limitations across laboratories that could hinder their global application to immune monitoring. In this methodological analysis, we compare protocols and test available reagents to identify sources of variability and to determine the limitations of the tetramer binding assay. We describe a robust pMHCII flow cytometry-based assay to quantify and phenotype antigen-specific CD4+ T cells directly ex vivo from frozen peripheral blood mononuclear cell samples, which we suggest should be tested across various laboratories to standardize immune-monitoring results.

  4. Flow cytometric analysis of peripheral blood and tumor-infiltrating regulatory T cells in dogs with oral malignant melanoma.

    Science.gov (United States)

    Tominaga, Makiko; Horiuchi, Yutaka; Ichikawa, Mika; Yamashita, Masao; Okano, Kumiko; Jikumaru, Yuri; Nariai, Yoko; Kadosawa, Tsuyoshi

    2010-05-01

    It is well known that tumor-infiltrating lymphocytes (TILs) and peripheral blood lymphocytes (PBLs) from patients with advanced-stage cancer have a poor immune response. Regulatory T cells (Tregs), characterized by the expression of a cluster of differentiation 4 and intracellular FoxP3 markers, can inhibit antitumor immunoresponse. In the present study, the prevalence of Tregs in peripheral blood and tumor tissue from dogs with oral malignant melanoma was evaluated by triple-color flow cytometry. The percentage of Tregs in the peripheral blood of the dogs with malignancy was significantly increased compared with healthy control dogs, and the percentage of Tregs within tumors was significantly increased compared with Tregs in peripheral blood of dogs with oral malignant melanoma. This finding suggests that the presence of tumor cells induced either local proliferation or selective migration of Tregs to tumor-infiltrated sites. A better understanding of the underlying mechanisms of Treg regulation in patients with cancer may lead to an effective anticancer immunotherapy against canine malignant melanoma and possibly other tumors.

  5. Immunophenotypic Characterization of Human Bone Marrow Mast Cells. A Flow Cytometric Study of Normal and Pathological Bone Marrow Samples

    Directory of Open Access Journals (Sweden)

    Luis Escribano

    1998-01-01

    Full Text Available The goal of the present paper was to define the immunophenotype of bone marrow mast cells (BMMC from healthy controls and patients with hematologic malignancies (HM based on the use of multiple stainings with monoclonal antibodies analyzed by flow cytometry. Our results show that BMMC from both groups of individuals display a similar but heterogenous immunophenotype. The overall numbers of BMMC are higher in the HM group of individuals (p = 0.08. Three patterns of antigen expression were detected: (1 markers constantly positive in all cases analyzed (CD9, CD29, CD33, CD43, CD44, CD49d, CD49e, CD51, CD71, CD117, and FcεRI, (2 antigens that were constantly negative (CD1a, CD2, CD3, CD5, CD6, CD11a, CD14, CD15, CD16, CD19, CD20, CD21, CD23, CD25, CD30, CD34, CD38, CD41a, CD42b, CD65, CD66b, HLA-DR, and CD138, and (3 markers that were positive in a variable proportion of cases – CD11b (50%, CD11c (77%, CD13 (40%, CD18 (20%, CD22 (68%, CD35 (27%, CD40 (67%, CD54 (88% and CD61 (40%. In addition, BMMC from all cases explored were CD45+, and this antigen was expressed at an intensity similar to that of mature granulocytes.

  6. Flow cytometric analysis of pollen grains collected from individual bees provides information about pollen load composition and foraging behaviour.

    Science.gov (United States)

    Kron, Paul; Kwok, Allison; Husband, Brian C

    2014-01-01

    Understanding the species composition of pollen on pollinators has applications in agriculture, conservation and evolutionary biology. Current identification methods, including morphological analysis, cannot always discriminate taxa at the species level. Recent advances in flow cytometry techniques for pollen grains allow rapid testing of large numbers of pollen grains for DNA content, potentially providing improved species resolution. A test was made as to whether pollen loads from single bees (honey-bees and bumble-bees) could be classified into types based on DNA content, and whether good estimates of proportions of different types could be made. An examination was also made of how readily DNA content can be used to identify specific pollen species. The method allowed DNA contents to be quickly found for between 250 and 9391 pollen grains (750-28 173 nuclei) from individual honey-bees and between 81 and 11 512 pollen grains (243-34 537 nuclei) for bumble-bees. It was possible to identify a minimum number of pollen species on each bee and to assign proportions of each pollen type (based on DNA content) present. The information provided by this technique is promising but is affected by the complexity of the pollination environment (i.e. number of flowering species present and extent of overlap in DNA content). Nevertheless, it provides a new tool for examining pollinator behaviour and between-species or cytotype pollen transfer, particularly when used in combination with other morphological, chemical or genetic techniques.

  7. Optimized multiparametric flow cytometric analysis of circulating endothelial cells and their subpopulations in peripheral blood of patients with solid tumors: a technical analysis

    Directory of Open Access Journals (Sweden)

    Zhou F

    2018-03-01

    Full Text Available Fangbin Zhou,1,2 Yaying Zhou,3 Ming Yang,1 Jinli Wen,3 Jun Dong,4 Wenyong Tan1 1Department of Oncology, The Second Clinical Medical College, Shenzhen People’s Hospital, Jinan University, Shenzhen, People’s Republic of China; 2Integrated Chinese and Western Medicine Postdoctoral Research Station, Jinan University, Guangzhou, People’s Republic of China; 3Clinical Medical Research Center, The Second Clinical Medical College, Shenzhen People’s Hospital, Jinan University, Shenzhen, People’s Republic of China; 4Department of Pathophysiology, Key Laboratory of the State Administration of Traditional Chinese Medicine, Medical College of Jinan University, Guangzhou, People’s Republic of China Background: Circulating endothelial cells (CECs and their subpopulations could be potential novel biomarkers for various malignancies. However, reliable enumerable methods are warranted to further improve their clinical utility. This study aimed to optimize a flow cytometric method (FCM assay for CECs and subpopulations in peripheral blood for patients with solid cancers.Patients and methods: An FCM assay was used to detect and identify CECs. A panel of 60 blood samples, including 44 metastatic cancer patients and 16 healthy controls, were used in this study. Some key issues of CEC enumeration, including sample material and anticoagulant selection, optimal titration of antibodies, lysis/wash procedures of blood sample preparation, conditions of sample storage, sufficient cell events to enhance the signal, fluorescence-minus-one controls instead of isotype controls to reduce background noise, optimal selection of cell surface markers, and evaluating the reproducibility of our method, were integrated and investigated. Wilcoxon and Mann–Whitney U tests were used to determine statistically significant differences.Results: In this validation study, we refined a five-color FCM method to detect CECs and their subpopulations in peripheral blood of patients

  8. Flow cytometric evaluation of the contribution of ionic silver to genotoxic potential of nanosilver in human liver HepG2 and colon Caco2 cells.

    Science.gov (United States)

    Sahu, Saura C; Njoroge, Joyce; Bryce, Steven M; Zheng, Jiwen; Ihrie, John

    2016-04-01

    Exposure to nanosilver found in food- and cosmetics-related consumer products is of public concern because of the lack of information about its safety. In this study, two widely used in vitro cell culture models, human liver HepG2 and colon Caco2 cells, and the flow cytometric micronucleus (FCMN) assay were evaluated as tools for rapid predictive screening of the potential genotoxicity of nanosilver. Recently, we reported the genotoxicity of 20 nm nanosilver using these systems. In the current study presented here, we tested the hypothesis that the nanoparticle size and cell types were critical determinants of its genotoxicity. To test this hypothesis, we used the FCMN assay to evaluate the genotoxic potential of 50 nm nanosilver of the same shape, composition, surface charge and obtained from the same commercial source using the same experimental conditions and in vitro models (HepG2 and Caco2) as previously tested for the 20 nm silver. Results of our study show that up to the concentrations tested in these cultured cell test systems, the smaller (20 nm) nanoparticle is genotoxic to both the cell types by inducing micronucleus (MN). However, the larger (50 nm) nanosilver induces MN only in HepG2 cells, but not in Caco2 cells. Also in this study, we evaluated the contribution of ionic silver to the genotoxic potential of nanosilver using silver acetate as the representative ionic silver. The MN frequencies in HepG2 and Caco2 cells exposed to the ionic silver in the concentration range tested are not statistically significant from the control values except at the top concentrations for both the cell types. Therefore, our results indicate that the ionic silver may not contribute to the MN-forming ability of nanosilver in HepG2 and Caco2 cells. Also our results suggest that the HepG2 and Caco2 cell cultures and the FCMN assay are useful tools for rapid predictive screening of a genotoxic potential of food- and cosmetics-related chemicals including nanosilver

  9. A novel and easy FxCycle™ violet based flow cytometric method for simultaneous assessment of DNA ploidy and six-color immunophenotyping.

    Science.gov (United States)

    Tembhare, Prashant; Badrinath, Yajamanam; Ghogale, Sitaram; Patkar, Nikhil; Dhole, Nilesh; Dalavi, Pooja; Kunder, Nikesh; Kumar, Ashok; Gujral, Sumeet; Subramanian, P G

    2016-03-01

    Abnormal DNA ploidy is a valuable prognostic factor in many neoplasms, especially in hematological neoplasms like B-cell acute lymphoblastic leukemia (B-ALL) and multiple myeloma (MM). Current methods of flow-cytometric (FC) DNA-ploidy evaluation are either technically difficult or limited to three- to four-color immunophenotyping and hence, challenging to evaluate DNA-ploidy in minute tumor population with background rich of its normal counterpart cells and other hematopoietic cells. We standardized a novel sensitive and easy method of simultaneous evaluation of six- to seven-color immunophenotyping and DNA-ploidy using a dye-FxCycle Violet (FCV). Linearity, resolution, and coefficient of variation (CV) for FCV were studied using chicken erythrocyte nuclei. Ploidy results of FCV were compared with Propidium iodide (PI) in 20 samples and intra-assay variation for FCV was studied. Using this six-color immunophenotyping & FCV-protocol DNA-ploidy was determined in bone-marrow samples from 124 B-ALL & 50 MM patients. Dilution experiment was also conducted to determine the sensitivity in detection of aneuploidy in minute tumor population. FCV revealed high linearity and resolution in 450/50 channel. On comparison with PI, CV of Go/G1-peak with FCV (mean-CV 4.1%) was slightly higher than PI (mean-CV 2.9%) but had complete agreement in ploidy results. Dilution experiment showed that aneuploidy could be accurately detected up to the limit of 0.01% tumor cells. Intra-assay variation was very low with CV of 0.005%. In B-ALL, hypodiploidy was noted in 4%, hyperdiploidy in 24%, near-hyperdiploidy in 13% and remaining 59% were diploid. In MM, hypodiploidy was in 2%, hyperdiploidy in 58%, near-hyperdiploidy in 8% and remaining 30% were diploid. FCV-based DNA-ploidy method is a sensitive and easy method for simultaneous evaluation of six-color immunophenotyping and DNA analysis. It is useful in DNA-ploidy evaluation of minute tumor population in cases like minimal residual

  10. Flow cytometric immunophenotyping of regulatory T cells in chronic lymphocytic leukemia: comparative assessment of various markers and use of novel antibody panel with CD127 as alternative to transcription factor FoxP3.

    Science.gov (United States)

    Dasgupta, Alakananda; Mahapatra, Manoranjan; Saxena, Renu

    2013-04-01

    This study analyzed the frequency of regulatory T cells (Tregs) in chronic lymphocytic leukemia (CLL) by multiparameter flow cytometric immunophenotyping. Patients showed significantly increased frequencies of Tregs as compared to controls, a significantly higher percentage than that identified by previous studies, possibly indicating a different prognosis of CLL in different parts of the world and, more precisely, a worse prognosis of CLL in the Indian population. A higher frequency of Tregs was also seen in advanced stage of disease with significantly reduced frequencies of Tregs in patients with CLL after chemotherapy. A significant proportion of CD127low/-FoxP3+ Tregs expressed only low levels of CD25. Thus, CD127 appears to be a better marker than CD25 for the identification of CD4+FoxP3+ T cells as potential Tregs. Our results suggest that the specificity and sensitivity of CD4+CD127low/- cells are comparable to those of CD4+FoxP3+, which is the gold standard, and can be used as an alternative. This novel flow cytometric antibody panel with fewer number of antibodies is cost-effective and can be used to enumerate Tregs in resource-limited settings.

  11. Cytometric analysis of mammalian sperm for induced morphologic and DNA content errors

    Energy Technology Data Exchange (ETDEWEB)

    Pinkel, D.

    1983-06-27

    Some flow-cytometric and image analysis procedures under development for quantitative analysis of sperm morphology are reviewed. The results of flow-cytometric DNA-content measurements on sperm from radiation exposed mice are also summarized, the results related to the available cytological information, and their potential dosimetric sensitivity discussed. (ACR)

  12. Cytometric analysis of mammalian sperm for induced morphologic and DNA content errors

    International Nuclear Information System (INIS)

    Pinkel, D.

    1983-01-01

    Some flow-cytometric and image analysis procedures under development for quantitative analysis of sperm morphology are reviewed. The results of flow-cytometric DNA-content measurements on sperm from radiation exposed mice are also summarized, the results related to the available cytological information, and their potential dosimetric sensitivity discussed

  13. Multiphase flow dynamics 5 nuclear thermal hydraulics

    CERN Document Server

    Kolev, Nikolay Ivanov

    2015-01-01

    This Volume 5 of the successful book package "Multiphase Flow Dynamics" is devoted to nuclear thermal hydraulics which is a substantial part of nuclear reactor safety. It provides knowledge and mathematical tools for adequate description of the process of transferring the fission heat released in materials due to nuclear reactions into its environment. It step by step introduces into the heat release inside the fuel, temperature fields in the fuels, the "simple" boiling flow in a pipe described using ideas of different complexity like equilibrium, non equilibrium, homogeneity, non homogeneity. Then the "simple" three-fluid boiling flow in a pipe is described by gradually involving the mechanisms like entrainment and deposition, dynamic fragmentation, collisions, coalescence, turbulence. All heat transfer mechanisms are introduced gradually discussing their uncertainty. Different techniques are introduced like boundary layer treatments or integral methods. Comparisons with experimental data at each step demons...

  14. Multiphase Flow Dynamics 5 Nuclear Thermal Hydraulics

    CERN Document Server

    Kolev, Nikolay Ivanov

    2012-01-01

    The present Volume 5 of the successful book package "Multiphase Flow Dynamics" is devoted to nuclear thermal hydraulics which is a substantial part of nuclear reactor safety. It provides knowledge and mathematical tools for adequate description of the process of transferring the fission heat released in materials due to nuclear reactions into its environment. It step by step introduces into the heat release inside the fuel, temperature fields in the fuels, the "simple" boiling flow in a pipe described using ideas of different complexity like equilibrium, non equilibrium, homogeneity, non homogeneity. Then the "simple" three-fluid boiling flow in a pipe is described by gradually involving the mechanisms like entrainment and deposition, dynamic fragmentation, collisions, coalescence, turbulence. All heat transfer mechanisms are introduced gradually discussing their uncertainty. Different techniques are introduced like boundary layer treatments or integral methods. Comparisons with experimental data at each step...

  15. An extended leukocyte differential count (16 types of circulating leukocytes) using the CytoDiff flow cytometric system can provide information for the discrimination of sepsis severity and prediction of outcome in sepsis patients.

    Science.gov (United States)

    Park, Sang Hyuk; Park, Borae G; Park, Chan-Jeoung; Kim, Sue; Kim, Duck-Hee; Jang, Seongsoo; Hong, Suk-Kyung; Chi, Hyun-Sook

    2014-07-01

    The Beckman Coulter CytoDiff flow cytometric system (Beckman Coulter, Miami, FL) was recently developed for performing leukocyte differential counts in up to 16 leukocyte subpopulations. We compared these leukocyte subpopulation levels among patients with three stages of sepsis (uncomplicated sepsis, severe sepsis, septic shock), especially focused on the discrimination of complicated sepsis from uncomplicated sepsis. We examined a total of 181 samples with sepsis who were admitted to the surgical intensive care unit. In addition, we examined samples obtained from 60 normal healthy volunteers. Both the proportions and absolute numbers of each cell type in the four groups were obtained using the CytoDiff flow cytometric system and compared. Mature neutrophils and immature granulocytes failed to discriminate patients with complicated sepsis from those with uncomplicated sepsis although their absolute numbers were increased compared with normal controls. In contrast, almost all lymphocyte subpopulations and CD16(-) monocytes decreased significantly in patients with complicated sepsis compared with uncomplicated sepsis. Among them, only B lymphocytes showed independent ability to discriminate two groups. Both B lymphocytes and CD16(-) monocytes possessed a significant adverse prognostic impact on overall survival when their absolute numbers decreased. Almost all lymphocyte subpopulations and CD16(-) monocytes decrease in size with increasing sepsis severity. Among them, only B lymphocytes showed independent ability to discriminate patients with complicated sepsis from those with uncomplicated sepsis. Both B lymphocytes and CD16 (-) monocytes show a significant adverse prognostic impact on overall survival outcomes in sepsis patients when their absolute numbers are decreased. Copyright © 2013 Clinical Cytometry Society.

  16. An extended leukocyte differential count (16 types of circulating leukocytes) using the cytodiff flow cytometric system can provide informations for the discrimination of sepsis severity and prediction of outcome in sepsis patients.

    Science.gov (United States)

    Park, Sang Hyuk; Park, Borae G; Park, Chan-Jeoung; Kim, Sue; Kim, Duck-Hee; Jang, Seongsoo; Hong, Suk-Kyung; Chi, Hyun-Sook

    2013-08-20

    Background: The Beckman Coulter CytoDiff flow cytometric system (Beckman Coulter, Miami, FL, USA) was recently developed for performing leukocyte differential counts in up to 16 leukocyte subpopulations. We compared these leukocyte subpopulation levels among patients with three stages of sepsis (uncomplicated sepsis, severe sepsis, septic shock), especially focused on the discrimination of complicated sepsis from uncomplicated sepsis. Methods: We examined a total of 181 samples with sepsis who were admitted to the surgical intensive care unit. In addition, we examined samples obtained from 60 normal healthy volunteers. Both the proportions and absolute numbers of each cell type in the four groups were obtained using the CytoDiff flow cytometric system and compared. Results: Mature neutrophils and immature granulocytes failed to discriminate patients with complicated sepsis from those with uncomplicated sepsis although their absolute numbers were increased compared with normal controls. In contrast, almost all lymphocyte subpopulations and CD16(-) monocytes decreased significantly in patients with complicated sepsis compared with uncomplicated sepsis. Among them, only B lymphocytes showed independent ability to discriminate two groups. Both B lymphocytes and CD16(-) monocytes possessed a significant adverse prognostic impact on overall survival when their absolute numbers decreased. Conclusions: Almost all lymphocyte subpopulations and CD16(-) monocytes decrease in size with increasing sepsis severity. Among them, only B lymphocytes showed independent ability to discriminate patients with complicated sepsis from those with uncomplicated sepsis. Both B lymphocytes and CD16(-) monocytes show a significant adverse prognostic impact on overall survival outcomes in sepsis patients when their absolute numbers are decreased. © 2013 Clinical Cytometry Society. Copyright © 2013 Clinical Cytometry Society.

  17. The nuclear safeguards data flow for the item facilities

    International Nuclear Information System (INIS)

    Wang Hongjun; Chen Desheng

    1994-04-01

    The constitution of nuclear safeguards data flow for the item facilities is introduced and the main contents are the data flow of nuclear safeguards. If the data flow moves positively, i.e. from source data →supporting documents→accounting records→accounting reports, the systems of records and reports will be constituted. If the data flow moves negatively, the way to trace inspection of nuclear material accounting quality will be constituted

  18. Nuclear engine flow reactivity shim control

    Science.gov (United States)

    Walsh, J.M.

    1973-12-11

    A nuclear engine control system is provided which automatically compensates for reactor reactivity uncertainties at the start of life and reactivity losses due to core corrosion during the reactor life in gas-cooled reactors. The coolant gas flow is varied automatically by means of specially provided control apparatus so that the reactor control drums maintain a predetermined steady state position throughout the reactor life. This permits the reactor to be designed for a constant drum position and results in a desirable, relatively flat temperature profile across the core. (Official Gazette)

  19. Flow cytometric sensitivity and characteristics of plasma cells in patients with multiple myeloma or its precursor disease: influence of biopsy site and anticoagulation method.

    Science.gov (United States)

    Manasanch, Elisabet E; Salem, Dalia A; Yuan, Constance M; Tageja, Nishant; Bhutani, Manisha; Kwok, Mary; Kazandjian, Dickran; Carter, George; Steinberg, Seth M; Zuchlinski, Diamond; Mulquin, Marcia; Calvo, Katherine; Maric, Irina; Roschewski, Mark; Korde, Neha; Braylan, Raul; Landgren, Ola; Stetler-Stevenson, Maryalice

    2015-05-01

    Flow cytometry has increasing relevance for prognosis in myeloma and precursor disease (monoclonal gammopathy of unknown significance/smoldering myeloma), yet it has been reported that plasma cell enumeration by flow varies depending on the quality of marrow aspirate and field biopsied in patchy disease. We demonstrated increased sensitivity of flow over immunohistochemistry in abnormal-plasma cell detection in monoclonal gammopathy (n = 59)/smoldering myeloma (n = 87). We prospectively evaluated treatment-na ve smoldering myeloma (n = 9)/myeloma (n = 11) patients for the percentage of abnormal plasma cells/total plasma cell compartment, plasma cell viability/infiltration and flow immunophenotype depending on anticoagulant use, biopsy site and pull sequence in uni-and-bilateral bone marrow biopsies and aspirates. We found no statistical difference regarding the percentage of abnormal plasma cells, their immunophenotype or number/distribution in marrow samples even when obtained by different sequence in aspirates, or anticoagulants (p > 0.05). Our results show that plasma cell enumeration and immunophenotyping by flow cytometry is consistent under different conditions in these populations.

  20. Flow cytometric readout based on Mitotracker Red CMXRos staining of live asexual blood stage malarial parasites reliably assesses antibody dependent cellular inhibition

    DEFF Research Database (Denmark)

    Jogdand, Prajakta S; Singh, Susheel K; Christiansen, Michael

    2012-01-01

    asynchronous and tightly synchronized asexual blood stage cultures of Plasmodium falciparum were stained with CMXRos and subjected to detection by flow cytometry and fluorescence microscopy. The parasite counts obtained by flow cytometry were compared to standard microscopic counts obtained through examination......ABSTRACT: BACKGROUND: Functional in vitro assays could provide insights into the efficacy of malaria vaccine candidates. For estimating the anti-parasite effect induced by a vaccine candidate, an accurate determination of live parasite count is an essential component of most in vitro bioassays....... Although traditionally parasites are counted microscopically, a faster, more accurate and less subjective method for counting parasites is desirable. In this study mitochondrial dye (Mitotracker Red CMXRos) was used for obtaining reliable live parasite counts through flow cytometry. METHODS: Both...

  1. Flow Cytometric Analysis of the Expression Pattern of Peroxisomal Proteins, Abcd1, Abcd2, and Abcd3 in BV-2 Murine Microglial Cells.

    Science.gov (United States)

    Debbabi, Meryam; Nury, Thomas; Helali, Imen; Karym, El Mostafa; Geillon, Flore; Gondcaille, Catherine; Trompier, Doriane; Najid, Amina; Terreau, Sébastien; Bezine, Maryem; Zarrouk, Amira; Vejux, Anne; Andreoletti, Pierre; Cherkaoui-Malki, Mustapha; Savary, Stéphane; Lizard, Gérard

    2017-01-01

    Microglial cells play important roles in neurodegenerative diseases including peroxisomal leukodystrophies. The BV-2 murine immortalized cells are widely used in the context of neurodegenerative researches. It is therefore important to establish the expression pattern of peroxisomal proteins by flow cytometry in these cells. So, the expression pattern of various peroxisomal transporters (Abcd1, Abcd2, Abcd3) contributing to peroxisomal β-oxidation was evaluated on BV-2 cells by flow cytometry and complementary methods (fluorescence microscopy, and RT-qPCR). By flow cytometry a strong expression of peroxisomal proteins (Abcd1, Abcd2, Abcd3) was observed. These data were in agreement with those obtained by fluorescence microscopy (presence of numerous fluorescent dots in the cytoplasm characteristic of a peroxisomal staining pattern) and RT-qPCR (high levels of Abcd1, Abcd2, and Abcd3 mRNAs). Thus, the peroxisomal proteins (Abcd1, Abcd2, Abcd3) are expressed in BV-2 cells, and can be analyzed by flow cytometry.

  2. Improving the resolution of cryopreserved X- and Y-sperm during DNA flow cytometric analysis with the addition of Percoll to quench the fluorescence of dead sperm

    NARCIS (Netherlands)

    Stap, J.; Hoebe, R. A.; Merton, J. S.; Haring, R. M.; Bakker, P. J.; Aten, J. A.

    1998-01-01

    The most effective method to control the sex of offspring is by separating X- from Y-bearing sperm on the basis of their DNA content. Sperm can be stained with Hoechst 33342 and efficiently sexed using a flow cytometer/cell sorter. However, applying this established assay to cryopreserved bovine

  3. Colour-encoded paramagnetic microbead-based direct inhibition triplex flow cytometric immunoassay for ochratoxin A, fumonisins and zearalenone in cereals and cereal-based feed

    NARCIS (Netherlands)

    Peters, J.; Thomas, D.; Boers, E.A.M.; Rijk, de T.C.; Berthiller, F.; Haasnoot, W.; Nielen, M.W.F.

    2013-01-01

    A combined (triplex) immunoassay for the simultaneous detection of three mycotoxins in grains was developed with superparamagnetic colour-encoded microbeads, in combination with two bead-dedicated flow cytometers. Monoclonal antibodies were coupled to the beads, and the amounts of bound mycotoxins

  4. High-Throughput Flow Cytometric Method for the Simultaneous Measurement of CAR-T Cell Characterization and Cytotoxicity against Solid Tumor Cell Lines.

    Science.gov (United States)

    Martinez, Emily M; Klebanoff, Samuel D; Secrest, Stephanie; Romain, Gabrielle; Haile, Samuel T; Emtage, Peter C R; Gilbert, Amy E

    2018-04-01

    High-throughput flow cytometry is an attractive platform for the analysis of adoptive cellular therapies such as chimeric antigen receptor T cell therapy (CAR-T) because it allows for the concurrent measurement of T cell-dependent cellular cytotoxicity (TDCC) and the functional characterization of engineered T cells with respect to percentage of CAR transduction, T cell phenotype, and measurement of T cell function such as activation in a single assay. The use of adherent tumor cell lines can be challenging in these flow-based assays. Here, we present the development of a high-throughput flow-based assay to measure TDCC for a CAR-T construct co-cultured with multiple adherent tumor cell lines. We describe optimal assay conditions (such as adherent cell dissociation techniques to minimize impact on cell viability) that result in robust cytotoxicity assays. In addition, we report on the concurrent use of T cell transduction and activation antibody panels (CD25) that provide further dissection of engineered T cell function. In conclusion, we present the development of a high-throughput flow cytometry method allowing for in vitro interrogation of solid tumor, targeting CAR-T cell-mediated cytotoxicity, CAR transduction, and engineered T cell characterization in a single assay.

  5. Flow cytometric readout based on Mitotracker Red CMXRos staining of live asexual blood stage malarial parasites reliably assesses antibody dependent cellular inhibition

    Directory of Open Access Journals (Sweden)

    Jogdand Prajakta S

    2012-07-01

    Full Text Available Abstract Background Functional in vitro assays could provide insights into the efficacy of malaria vaccine candidates. For estimating the anti-parasite effect induced by a vaccine candidate, an accurate determination of live parasite count is an essential component of most in vitro bioassays. Although traditionally parasites are counted microscopically, a faster, more accurate and less subjective method for counting parasites is desirable. In this study mitochondrial dye (Mitotracker Red CMXRos was used for obtaining reliable live parasite counts through flow cytometry. Methods Both asynchronous and tightly synchronized asexual blood stage cultures of Plasmodium falciparum were stained with CMXRos and subjected to detection by flow cytometry and fluorescence microscopy. The parasite counts obtained by flow cytometry were compared to standard microscopic counts obtained through examination of Giemsa-stained thin smears. A comparison of the ability of CMXRos to stain live and compromised parasites (induced by either medium starvation or by anti-malarial drug treatment was carried out. Finally, parasite counts obtained by CMXRos staining through flow cytometry were used to determine specific growth inhibition index (SGI in an antibody-dependent cellular inhibition (ADCI assay. Results Mitotracker Red CMXRos can reliably detect live intra-erythrocytic stages of P. falciparum. Comparison between staining of live with compromised parasites shows that CMXRos predominantly stains live parasites with functional mitochondria. Parasite counts obtained by CMXRos staining and flow cytometry were highly reproducible and can reliably determine the ability of IgG from hyper-immune individuals to inhibit parasite growth in presence of monocytes in ADCI assay. Further, a dose-dependent parasite growth inhibitory effect could be detected for both total IgG purified from hyper-immune sera and affinity purified IgGs against the N-terminal non-repeat region of GLURP

  6. Flow cytometric readout based on Mitotracker Red CMXRos staining of live asexual blood stage malarial parasites reliably assesses antibody dependent cellular inhibition.

    Science.gov (United States)

    Jogdand, Prajakta S; Singh, Susheel K; Christiansen, Michael; Dziegiel, Morten H; Singh, Subhash; Theisen, Michael

    2012-07-20

    Functional in vitro assays could provide insights into the efficacy of malaria vaccine candidates. For estimating the anti-parasite effect induced by a vaccine candidate, an accurate determination of live parasite count is an essential component of most in vitro bioassays. Although traditionally parasites are counted microscopically, a faster, more accurate and less subjective method for counting parasites is desirable. In this study mitochondrial dye (Mitotracker Red CMXRos) was used for obtaining reliable live parasite counts through flow cytometry. Both asynchronous and tightly synchronized asexual blood stage cultures of Plasmodium falciparum were stained with CMXRos and subjected to detection by flow cytometry and fluorescence microscopy. The parasite counts obtained by flow cytometry were compared to standard microscopic counts obtained through examination of Giemsa-stained thin smears. A comparison of the ability of CMXRos to stain live and compromised parasites (induced by either medium starvation or by anti-malarial drug treatment) was carried out. Finally, parasite counts obtained by CMXRos staining through flow cytometry were used to determine specific growth inhibition index (SGI) in an antibody-dependent cellular inhibition (ADCI) assay. Mitotracker Red CMXRos can reliably detect live intra-erythrocytic stages of P. falciparum. Comparison between staining of live with compromised parasites shows that CMXRos predominantly stains live parasites with functional mitochondria. Parasite counts obtained by CMXRos staining and flow cytometry were highly reproducible and can reliably determine the ability of IgG from hyper-immune individuals to inhibit parasite growth in presence of monocytes in ADCI assay. Further, a dose-dependent parasite growth inhibitory effect could be detected for both total IgG purified from hyper-immune sera and affinity purified IgGs against the N-terminal non-repeat region of GLURP in ADCI assays coupled with determination of

  7. Exploring the feasibility of multi-site flow cytometric processing of gut associated lymphoid tissue with centralized data analysis for multi-site clinical trials.

    Science.gov (United States)

    McGowan, Ian; Anton, Peter A; Elliott, Julie; Cranston, Ross D; Duffill, Kathryn; Althouse, Andrew D; Hawkins, Kevin L; De Rosa, Stephen C

    2015-01-01

    The purpose of this study was to determine whether the development of a standardized approach to the collection of intestinal tissue from healthy volunteers, isolation of gut associated lymphoid tissue mucosal mononuclear cells (MMC), and characterization of mucosal T cell phenotypes by flow cytometry was sufficient to minimize differences in the normative ranges of flow parameters generated at two trial sites. Forty healthy male study participants were enrolled in Pittsburgh and Los Angeles. MMC were isolated from rectal biopsies using the same biopsy acquisition and enzymatic digestion protocols. As an additional comparator, peripheral blood mononuclear cells (PBMC) were collected from the study participants. For quality control, cryopreserved PBMC from a single donor were supplied to both sites from a central repository (qPBMC). Using a jointly optimized standard operating procedure, cells were isolated from tissue and blood and stained with monoclonal antibodies targeted to T cell phenotypic markers. Site-specific flow data were analyzed by an independent center which analyzed all data from both sites. Ranges for frequencies for overall CD4+ and CD8+ T cells, derived from the qPBMC samples, were equivalent at both UCLA and MWRI. However, there were significant differences across sites for the majority of T cell activation and memory subsets in qPBMC as well as PBMC and MMC. Standardized protocols to collect, stain, and analyze MMC and PBMC, including centralized analysis, can reduce but not exclude variability in reporting flow data within multi-site studies. Based on these data, centralized processing, flow cytometry, and analysis of samples may provide more robust data across multi-site studies. Centralized processing requires either shipping of fresh samples or cryopreservation and the decision to perform centralized versus site processing needs to take into account the drawbacks and restrictions associated with each method.

  8. Exploring the feasibility of multi-site flow cytometric processing of gut associated lymphoid tissue with centralized data analysis for multi-site clinical trials.

    Directory of Open Access Journals (Sweden)

    Ian McGowan

    Full Text Available The purpose of this study was to determine whether the development of a standardized approach to the collection of intestinal tissue from healthy volunteers, isolation of gut associated lymphoid tissue mucosal mononuclear cells (MMC, and characterization of mucosal T cell phenotypes by flow cytometry was sufficient to minimize differences in the normative ranges of flow parameters generated at two trial sites. Forty healthy male study participants were enrolled in Pittsburgh and Los Angeles. MMC were isolated from rectal biopsies using the same biopsy acquisition and enzymatic digestion protocols. As an additional comparator, peripheral blood mononuclear cells (PBMC were collected from the study participants. For quality control, cryopreserved PBMC from a single donor were supplied to both sites from a central repository (qPBMC. Using a jointly optimized standard operating procedure, cells were isolated from tissue and blood and stained with monoclonal antibodies targeted to T cell phenotypic markers. Site-specific flow data were analyzed by an independent center which analyzed all data from both sites. Ranges for frequencies for overall CD4+ and CD8+ T cells, derived from the qPBMC samples, were equivalent at both UCLA and MWRI. However, there were significant differences across sites for the majority of T cell activation and memory subsets in qPBMC as well as PBMC and MMC. Standardized protocols to collect, stain, and analyze MMC and PBMC, including centralized analysis, can reduce but not exclude variability in reporting flow data within multi-site studies. Based on these data, centralized processing, flow cytometry, and analysis of samples may provide more robust data across multi-site studies. Centralized processing requires either shipping of fresh samples or cryopreservation and the decision to perform centralized versus site processing needs to take into account the drawbacks and restrictions associated with each method.

  9. Corrigendum to "Flow cytometric quantitation of platelet phagocytosis by monocytes using a pH-sensitive dye, pHrodo-SE" [Journal of Immunological Methods 447 (2017) 57-64].

    Science.gov (United States)

    Takahashi, Daisuke; Fujihara, Mitsuhiro; Miyazaki, Toru; Matsubayashi, Keiji; Sato, Shinichiro; Azuma, Hiroshi; Kato, Toshiaki; Kino, Shuichi; Ikeda, Hisami; Takamoto, Shigeru; Sato, Noriyuki; Torigoe, Toshihiko

    2018-03-01

    Antibody-mediated phagocytosis of platelets using a flow cytometric monocyte-based phagocytosis assay (FMPA) has been shown to predict the outcome of platelet transfusion. The easy adherence between platelets and monocytes even in the absence of an antibody is regarded as one of limitations of the FMPA. To improve the FMPA for prediction of transfusion outcome, we used the pH-sensitive dye pHrodo succinimidyl ester (pHrodo-SE), which has weak fluorescence at neutral pH and has increased fluorescence intensity in low pH conditions such as in lysomes. Platelets stained with pHrodo-SE were sensitized with an HLA class I monoclonal antibody (w6/32 clone) or anti-HLA class I containing antisera. The platelets were incubated with monocyte-enriched mononuclear cells. Phagocytic activity was assessed by the percentage of monocytes that phagocytosed platelets. Sensitization of platelets with w6/32 significantly increased platelet phagocytosis by monocytes in dose- and time-dependent manners. Anti-HLA class I antibody-containing sera caused platelet phagocytosis in a cognate antigen-antibody-dependent manner. There was a significant correlation (r=0.69, p<0.01) between phagocytic index and titer of HLA class I antibody measured by lymphocyte immunofluorescence test-flow cytometry. In addition, the phagocytic index obtained by FMPA with pHrodo-SE was significantly higher than that obtained by FMPA with the previously used dye, carboxyfluorescein diacetate succinimidyl ester, when platelets were sensitized by w6/32 and anti-HLA class I antibody-containing sera. Because of the higher resolution and higher sensitivity than those of the previous method, the pHrodo-SE-based FMPA may be suitable for more precise quantitation of phagocytosis activity, which would enable qualitative evaluation of transfusion effectiveness. Copyright © 2018.

  10. Flow Cytometric DNA index, G-band Karyotyping, and Comparative Genomic Hybridization in Detection of High Hyperdiploidy in Childhood Acute Lymphoblastic Leukemia

    DEFF Research Database (Denmark)

    Nygaard, Ulrikka; Larsen, Jacob; Kristensen, Tim D

    2006-01-01

    High hyperdiploid acute lymphoblastic leukemia in children is related to a good outcome. Because these patients may be stratified to a low-intensity treatment, we have investigated the sensitivity of flow cytometry (FCM), G-band karyotyping (GBK), and high-resolution comparative genomic hybridiza......High hyperdiploid acute lymphoblastic leukemia in children is related to a good outcome. Because these patients may be stratified to a low-intensity treatment, we have investigated the sensitivity of flow cytometry (FCM), G-band karyotyping (GBK), and high-resolution comparative genomic...... hybridization (HR-CGH) in detecting high hyperdiploid leukemic clones. Twenty-six girls and 34 boys with acute lymphoblastic leukemia diagnosed in 1998 to 1999 were analyzed by FCM, GBK, and HR-CGH. The correlations between DNA indices obtained by FCM, GBK, and HR-CGH were significant (rs=0.61 to 0.77; P

  11. Flow cytometric applications of tumor biology: prospects and pitfalls. [Applications in study of spontaneous dog tumors and in drug and radiation effects on cultured V79 cells

    Energy Technology Data Exchange (ETDEWEB)

    Raju, M.R.; Johnson, T.S.; Tokita, N.; Gillette, E.L.

    1979-01-01

    A brief review of cytometry instrumentation and its potential applications in tumor biology is presented using our recent data. Age-distribution measurements of cells from spontaneous dog tumors and cultured cells after exposure to x rays, alpha particles, or adriamycin are shown. The data show that DNA fluorescence measurements have application in the study of cell kinetics after either radiation or drug treatment. Extensive and careful experimentation is needed to utilize the sophisticated developments in flow cytometry instrumentation.

  12. In-depth validation of acridine orange staining for flow cytometric parasite and reticulocyte enumeration in an experimental model using Plasmodium berghei

    DEFF Research Database (Denmark)

    Hein-Kristensen, L; Wiese, L; Kurtzhals, J A L

    2009-01-01

    Flow cytometry is potentially an effective method for counting malaria parasites, but inconsistent results have hampered its routine use in rodent models. A published two-channel method using acridine orange offers clear discrimination between the infected and uninfected erythrocytes. However......, preliminary studies showed concerns when dealing with Plasmodium berghei-infected blood samples with high numbers of reticulocytes. In hyperparasitemic or chronic P. berghei infection, enhanced erythropoietic activity results in high numbers of circulating immature reticulocytes. We show that even though...

  13. Super-resolved calibration-free flow cytometric characterization of platelets and cell-derived microparticles in platelet-rich plasma.

    Science.gov (United States)

    Konokhova, Anastasiya I; Chernova, Darya N; Moskalensky, Alexander E; Strokotov, Dmitry I; Yurkin, Maxim A; Chernyshev, Andrei V; Maltsev, Valeri P

    2016-02-01

    Importance of microparticles (MPs), also regarded as extracellular vesicles, in many physiological processes and clinical conditions motivates one to use the most informative and precise methods for their characterization. Methods based on individual particle analysis provide statistically reliable distributions of MP population over characteristics. Although flow cytometry is one of the most powerful technologies of this type, the standard forward-versus-side-scattering plots of MPs and platelets (PLTs) overlap considerably because of similarity of their morphological characteristics. Moreover, ordinary flow cytometry is not capable of measurement of size and refractive index (RI) of MPs. In this study, we 1) employed the potential of the scanning flow cytometer (SFC) for identification and characterization of MPs from light scattering; 2) suggested the reference method to characterize MP morphology (size and RI) with high precision; and 3) determined the lowest size of a MP that can be characterized from light scattering with the SFC. We equipped the SFC with 405 and 488 nm lasers to measure the light-scattering profiles and side scattering from MPs, respectively. The developed two-stage method allowed accurate separation of PLTs and MPs in platelet-rich plasma. We used two optical models for MPs, a sphere and a bisphere, in the solution of the inverse light-scattering problem. This solution provides unprecedented precision in determination of size and RI of individual spherical MPs-median uncertainties (standard deviations) were 6 nm and 0.003, respectively. The developed method provides instrument-independent quantitative information on MPs, which can be used in studies of various factors affecting MP population. © 2015 International Society for Advancement of Cytometry.

  14. Flow Cytometric DNA index, G-band Karyotyping, and Comparative Genomic Hybridization in Detection of High Hyperdiploidy in Childhood Acute Lymphoblastic Leukemia

    DEFF Research Database (Denmark)

    Nygaard, Ulrikka; Larsen, Jacob; Kristensen, Tim D

    2006-01-01

    High hyperdiploid acute lymphoblastic leukemia in children is related to a good outcome. Because these patients may be stratified to a low-intensity treatment, we have investigated the sensitivity of flow cytometry (FCM), G-band karyotyping (GBK), and high-resolution comparative genomic hybridiza......High hyperdiploid acute lymphoblastic leukemia in children is related to a good outcome. Because these patients may be stratified to a low-intensity treatment, we have investigated the sensitivity of flow cytometry (FCM), G-band karyotyping (GBK), and high-resolution comparative genomic.......001 for all comparisons). However, in 4 of 18 patients, high hyperdiploidy was overlooked by GBK or HR-CGH, and even when FCM was applied, 2 of 18 patients with high hyperdiploidy by GBK and/or HR-CGH were classified as nonhigh hyperdiploid. If high hyperdiploid subclones were included, FCM could detect all...... high hyperdiploid patients found by either GBK or HR-CGH, but would then in addition classify 15% to 20% of the remaining patients as high hyperdiploid. Thus, both GBK and HR-CGH overlook patients with high hyperdiploidy, and FCM only detects all high hyperdiploid patients if small high hyperdiploid...

  15. Identification of microbes from the surfaces of food-processing lines based on the flow cytometric evaluation of cellular metabolic activity combined with cell sorting.

    Science.gov (United States)

    Juzwa, W; Duber, A; Myszka, K; Białas, W; Czaczyk, K

    2016-09-01

    In this study the design of a flow cytometry-based procedure to facilitate the detection of adherent bacteria from food-processing surfaces was evaluated. The measurement of the cellular redox potential (CRP) of microbial cells was combined with cell sorting for the identification of microorganisms. The procedure enhanced live/dead cell discrimination owing to the measurement of the cell physiology. The microbial contamination of the surface of a stainless steel conveyor used to process button mushrooms was evaluated in three independent experiments. The flow cytometry procedure provided a step towards monitoring of contamination and enabled the assessment of microbial food safety hazards by the discrimination of active, mid-active and non-active bacterial sub-populations based on determination of their cellular vitality and subsequently single cell sorting to isolate microbial strains from discriminated sub-populations. There was a significant correlation (r = 0.97; p vitality and the identification of species from defined sub-populations, although the identified microbes were limited to culturable cells.

  16. Assessment of the viability and fertilizing potential of cryopreserved bovine spermatozoa using dual fluorescent staining and two-flow cytometric systems.

    Science.gov (United States)

    Ericsson, S A; Garner, D L; Redelman, D; Ahmad, K

    1989-04-01

    A dual fluorescent staining system utilizing 5 (and-6)-carboxy-4',5'-dimethyl fluorescein diacetate (CDMFDA) and Hydroethidine (HED) was developed to provide quantifiable information reflective of spermatozoal viability and fertilizing potential. Cryopreserved spermatozoa from ten bulls on which there was fertilizing capacity information were incubated for 1.5, and 3 hr at 39 degrees C prior to fluorogenic staining. Spermatozoa were analyzed using both a FACS Analyzer and an EPICS V flow cytometer to determine if a particular fluorescence pattern was due to an instrumental artifact or cellular processes. Five fluorescent cellular populations were identified by the FACS Analyzer and three populations by the EPICS V. Spermatozoa were quantified after each incubation time for red (HED) and green (CDMFDA) fluorescence. Viable spermatozoa retained the greatest amount of both green and red fluorescence. Dead or moribund spermatozoa had a decrease in over-all fluorescence. The number of viable cells at 0 hr plus the number of dead or morbid cells at any time period were identified by the FACS Analyzer as important in estimating the potential fertility of a bull. The EPICS V identified the number of dead or moribund cells as being related to nonreturn rates. Incubation of samples decreased cellular viability, which resulted in reduced levels of both green and red fluorescence. Similarities between data obtained with both flow cytometers illustrated that cellular processes, not instrumental artifacts, were responsible for the decrease in over-all fluorescence when viability declined, the relationship between the number of cells with specific fluorescence levels and nonreturn rates, and the incubative-induced changes in fluorescence patterns.

  17. Flow-cytometric measurement of CD4-8- T cells bearing T-cell receptor αβ chains, 1

    International Nuclear Information System (INIS)

    Kusunoki, Yoichiro; Hirai, Yuko; Kyoizumi, Seishi; Akiyama, Mitoshi.

    1992-09-01

    In this study we detected rare, possibly abnormal, T cells bearing CD3 surface antigen and T-cell receptor (TCR) αβ chains but lacking both CD4 and CD8 antigens (viz., TCRαβ + CD4 - 8 - cells, as determined by flow cytometry). The TCRαβ + CD4 - 8 - T cells were detected at a mean frequency of 0.63 ± 0.35 % (mean ± standard deviation) in peripheral blood TCRαβ + cells of 119 normal persons. Two unusual cases besides the 119 normal persons showed extremely elevated frequencies of TCRαβ + CD4 - 8 - T cells, viz., approximately 5 % to 10 % and 14 % to 19 % in whole TCRαβ + cells. Both individuals were males who were otherwise physiologically quite normal with no history of severe illness, and these high frequencies were also observed in blood samples collected 2 or 8 years prior to the current measurements. The TCRαβ + CD4 - 8 - T cells of the two individuals were found to express mature T-cell markers such as CD2,3, and 5 antigens, as well as natural killer (NK) cell markers, viz., CD11b, 16, 56, and 57 antigens, when peripheral blood lymphocytes were subjected to three-color flow cytometry. Lectin-dependent or redirected antibody-dependent cell-mediated cytotoxicities were observed for both freshly sorted TCRαβ + CD4 - 8 - cells and in vitro established clones. Nevertheless, NK-like activity was not detected. Further, Southern blot analysis of TCRβ and γ genes revealed identical rearrangement patterns for all the TCRαβ + CD4 - 8 - clones established in vitro. These results suggest that the TCRαβ + CD4 - 8 - T cells from these two mean exhibit unique characteristics and proliferate clonally in vivo. (author)

  18. Lymphocyte subsets show different response patterns to in vivo bound natalizumab--a flow cytometric study on patients with multiple sclerosis.

    Directory of Open Access Journals (Sweden)

    Andrea Harrer

    Full Text Available Natalizumab is an effective monoclonal antibody therapy for the treatment of relapsing-remitting multiple sclerosis (RRMS and interferes with immune cell migration into the central nervous system by blocking the α(4 subunit of very-late activation antigen-4 (VLA-4. Although well tolerated and very effective, some patients still suffer from relapses in spite of natalizumab therapy or from unwanted side effects like progressive multifocal leukoencephalopathy (PML. In search of a routine-qualified biomarker on the effectiveness of natalizumab therapy we applied flow cytometry and analyzed natalizumab binding to α(4 and α(4 integrin surface levels on T-cells, B-cells, natural killer (NK cells, and NKT cells from 26 RRMS patients under up to 72 weeks of therapy. Four-weekly infusions of natalizumab resulted in a significant and sustained increase of lymphocyte-bound natalizumab (p<0.001 which was paralleled by a significant decrease in detectability of the α(4 integrin subunit on all lymphocyte subsets (p<0.001. We observed pronounced natalizumab accumulations on T and B cells at single measurements in all patients who reported clinical disease activity (n = 4. The natalizumab binding capacity of in vitro saturated lymphocytes collected during therapy was strongly diminished compared to treatment-naive cells indicating a therapy-induced reduction of α(4. Summing up, this pilot study shows that flow cytometry is a useful method to monitor natalizumab binding to lymphocytes from RRMS patients under therapy. Investigating natalizumab binding provides an opportunity to evaluate the molecular level of effectiveness of natalizumab therapy in individual patients. In combination with natalizumab saturation experiments, it possibly even provides a means of studying the feasability of patient-tailored infusion intervals. A routine-qualified biomarker on the basis of individual natalizumab saturation on lymphocyte subsets might be an effective tool to

  19. Normal adult ramified microglia separated from other central nervous system macrophages by flow cytometric sorting: Phenotypic differences defined and direct ex vivo antigen presentation to myelin basic protein-reactive CD4{sup +} T cells compared

    Energy Technology Data Exchange (ETDEWEB)

    Ford, A.L.; Goodsall, A.L.; Sedgwick, J.D. [Centenary Institute of Cancer Medicine and Cell Biology, Sydney (Australia)] [and others

    1995-05-01

    Ramified microglia in the adult central nervous system (CNS) are the principal glial element up-regulating MHC class I and II expression in response to inflammatory events or neuronal damage. A proportion of these cells also express MHC class II constitutively in the normal CNS. The role of microglia as APCs for CD4{sup +} cells extravasating into the CNS remains undefined. In this study, using irradiation bone marrow chimeras in CD45-congenic rats, the phenotype CD45{sup low}CD11b/c{sup +} is shown to identify microglial cells specifically within the CNS. Highly purified populations of microglia and nonmicroglial but CNS-associated macrophages (CD45{sup high}CD11b/c{sup +}) have been obtained directly from the adult CNS, by using flow cytometric sorting. Morphologically, freshly isolated microglia vs other CNS macrophages are quite distinct. Of the two populations recovered from the normal CNS, it is the minority CD45{sup high}CD11 b/c{sup +} transitional macrophage population, and not microglia, that is the effective APC for experimental autoimmune encephalomyelitis-inducing CD4{sup +} myelin basic protein (MBP)-reactive T cells. CD45{sup high}CD11b/c{sup +} CNS macrophages also stimulate MBP-reactive T cells without addition of MBP to culture suggesting presentation of endogenous Ag. This is the first study in which microglia vs other CNS macrophages have been analyzed for APC ability directly from the CNS, with substantial cross-contamination between the two populations eliminated. The heterogeneity of these populations in terms of APC function is clearly demonstrated. Evidence is still lacking that adult CNS microglia have the capacity to interact with and stimulate CD4{sup +} T cells to proliferate or secrete IL-2. 60 refs., 6 figs., 1 tab.

  20. Innovations in diagnosis and post-therapeutic monitoring of Chagas disease: Simultaneous flow cytometric detection of IgG1 antibodies anti-live amastigote, anti-live trypomastigote, and anti-fixed epimastigote forms of Trypanosoma cruzi.

    Science.gov (United States)

    Alessio, Glaucia Diniz; Côrtes, Denise Fonseca; Machado de Assis, Girley Francisco; Júnior, Policarpo Ademar Sales; Ferro, Eloisa Amália Vieira; Antonelli, Lis Ribeiro do Valle; Teixeira-Carvalho, Andréa; Martins-Filho, Olindo Assis; de Lana, Marta

    2014-11-01

    This study developed a remarkable methodological innovation (FC-ATE) which enables simultaneous detection of antibodies specific to the three evolutive forms of Trypanosoma cruzi: live amastigote (AMA), live trypomastigote (TRYPO), and fixed epimastigote (EPI) using a differential fluorescence staining as low (AMA), intermediate (TRYPO), and high (EPI). An outstanding performance (100%) was observed in the discrimination of the chagasic (CH) and non-chagasic (NCH) patients. In the applicability of FC-ATE in the diagnosis of Chagas disease, 100% of the CH samples presented positivity in the percentage of positive fluorescent parasites (PPFP) for all the three forms of T. cruzi. Moreover, 94% of the samples of NCH presented negative values of PPFP with AMA and TRYPO, and 88% with EPI. Samples from the NCH group with false-positive results were those belonging to the leishmaniasis patients. Considering the applicability of this technique in post-therapeutic monitoring of Chagas disease, 100% of non-treated (NT) and treated non-cured (TNC) samples were positive with the three T. cruzi evolutive forms, while a percentage of 100% from samples of the treated cured (TC) patients were negative with AMA, 93% with TRYPO and 96% with EPI. The comparison between FC-ATE and two other flow cytometric tests using the same samples of patients NT, TNC and TC showed that the three techniques presented different reactivities, although categorical correlation between the methodologies was observed. Taken together, the results obtained with the novel FC-ATE method have shown an outstanding performance in the diagnosis and post-therapeutic monitoring of Chagas disease. Copyright © 2014 Elsevier B.V. All rights reserved.

  1. Further evaluation of the prognostic value of morphometric and flow cytometric parameters in breast-cancer patients with long follow-up.

    Science.gov (United States)

    Uyterlinde, A M; Baak, J P; Schipper, N W; Peterse, H; Matze, E; Meijer, C J

    1990-01-15

    The added prognostic value of cellular DNA content compared with single and combined morphometric factors and classical parameters such as tumor size, nodal status, histologic grade and estrogen receptor (ER) content was investigated in 225 consecutive breast-cancer patients with long follow-up. Of all features investigated, the MPI (multivariate prognostic index) had the strongest prognostic value [Mantel-Cox (MC) = 48.2, p less than 0.00005]. The results further showed that neither age nor ER content had significant prognostic value, but the DNA index (DI) as a single parameter had (though weak) prognostic significance (MC = 5.9, p = 0.015); a similar result was obtained with the percentage of S-phase cells (MC = 6.1, p = 0.013). The DI had (restricted) additional prognostic value to the morphometric features (MPI plus DI Mantel-Cox 53.0, p less than 0.0001). The percentage of S-phase cells had no additional prognostic value over the MPI. On the other hand, the additional value of the DI over tumor size and nodal status was much more impressive (MC = 41.0 and 40.7), although it did not reach the prognostic significance of the MPI. Prediction of disease outcome with a linear combination of quantitative microscopical parameters of the primary tumor alone [MAI (mitotic activity index), DI and mean nuclear area] was very accurate, even without considering lymph-node status (MC 30.8, p less than 0.0005). Grade had no additional value to the MPI at all (p = 0.76). This could be especially important for lymph-node-negative patients in whom the prognostic value of the MPI and the MAI are confirmed.

  2. Corrected Lymphocyte Percentages Reduce the Differences in Absolute CD4+ T Lymphocyte Counts between Dual-Platform and Single-Platform Flow Cytometric Approaches.

    Science.gov (United States)

    Noulsri, Egarit; Abudaya, Dinar; Lerdwana, Surada; Pattanapanyasat, Kovit

    2018-03-13

    To determine whether a corrected lymphocyte percentage could reduce bias in the absolute cluster of differentiation (CD)4+ T lymphocyte counts obtained via dual-platform (DP) vs standard single-platform (SP) flow cytometry. The correction factor (CF) for the lymphocyte percentages was calculated at 6 laboratories. The absolute CD4+ T lymphocyte counts in 300 blood specimens infected with human immunodeficiency virus (HIV) were determined using the DP and SP methods. Applying the CFs revealed that 4 sites showed a decrease in the mean bias of absolute CD4+ T lymphocyte counts determined via DP vs standard SP (-109 vs -84 cells/μL, -80 vs -58 cells/μL, -52 vs -45 cells/μL, and -32 vs 1 cells/μL). However, 2 participating laboratories revealed an increase in the difference of the mean bias (-42 vs -49 cells/μL and -20 vs -69 cells/μL). Use of the corrected lymphocyte percentage shows potential for decreasing the difference in CD4 counts between DP and the standard SP method.

  3. In vitro effects of zirconia and alumina particles on human blood monocyte-derived macrophages: X-ray microanalysis and flow cytometric studies.

    Science.gov (United States)

    Nkamgueu, E M; Adnet, J J; Bernard, J; Zierold, K; Kilian, L; Jallot, E; Benhayoune, H; Bonhomme, P

    2000-12-15

    The cytocompatibility of two particulate bioceramics, zirconia and alumina, was studied using human blood monocytes driven to differentiate into mature macrophages with granulocyte macrophage-colony-stimulating factor. Changes in individual cell elemental composition, particularly sodium and potassium content, were assessed by X-ray microanalysis of ultrathin freeze-dried sections. Phagocytosis and respiratory burst of macrophages exposed to biomaterial for 7 days were analyzed under flow cytometry using uptake of fluorescent latex beads and 2'7'-dichlorofluorescien diacetate oxidation, respectively. Zirconia and alumina particles were found to decrease the intracellular potassium/sodium ratio (an index of cell vitality) significantly (p2 times reduced by zirconia and >5 times reduced by alumina). The present study clearly demonstrates that reduction of the phagocytic capacity of macrophages associated with altered oxidative metabolism caused by biomaterial particles is characterized by changes in intracellular elemental content. Thus, investigation of cellular homeostasis by electron probe microanalysis together with analysis of functional changes may improve estimation of biomaterial cytocompatibility. Copyright 2000 John Wiley & Sons, Inc.

  4. Lymphocyte subsets show different response patterns to in vivo bound natalizumab--a flow cytometric study on patients with multiple sclerosis.

    Science.gov (United States)

    Harrer, Andrea; Pilz, Georg; Einhaeupl, Max; Oppermann, Katrin; Hitzl, Wolfgang; Wipfler, Peter; Sellner, Johann; Golaszewski, Stefan; Afazel, Shahrzad; Haschke-Becher, Elisabeth; Trinka, Eugen; Kraus, Joerg

    2012-01-01

    Natalizumab is an effective monoclonal antibody therapy for the treatment of relapsing-remitting multiple sclerosis (RRMS) and interferes with immune cell migration into the central nervous system by blocking the α(4) subunit of very-late activation antigen-4 (VLA-4). Although well tolerated and very effective, some patients still suffer from relapses in spite of natalizumab therapy or from unwanted side effects like progressive multifocal leukoencephalopathy (PML). In search of a routine-qualified biomarker on the effectiveness of natalizumab therapy we applied flow cytometry and analyzed natalizumab binding to α(4) and α(4) integrin surface levels on T-cells, B-cells, natural killer (NK) cells, and NKT cells from 26 RRMS patients under up to 72 weeks of therapy. Four-weekly infusions of natalizumab resulted in a significant and sustained increase of lymphocyte-bound natalizumab (pqualified biomarker on the basis of individual natalizumab saturation on lymphocyte subsets might be an effective tool to improve treatment safety.

  5. Flow Cytometric Determination of Cellular Sources and Frequencies of Key Cytokine-Producing Lymphocytes Directed against Recombinant LACK and Soluble Leishmania Antigen in Human Cutaneous Leishmaniasis

    Science.gov (United States)

    Bottrel, R. L. A.; Dutra, W. O.; Martins, F. A.; Gontijo, B.; Carvalho, E.; Barral-Netto, M.; Barral, A.; Almeida, R. P.; Mayrink, W.; Locksley, R.; Gollob, K. J.

    2001-01-01

    Leishmaniasis, caused by infection with the protozoan parasite Leishmania, affects millions of individuals worldwide, causing serious morbidity and mortality. This study directly determined the frequency of cells producing key immunoregulatory cytokines in response to the recombinant antigen Leishmania homolog of receptors for activated kinase C (LACK) and soluble leishmania antigen (SLA), and it determined relative contributions of these antigens to the overall cytokine profile in individuals infected for the first time with Leishmania braziliensis. All individuals presented with the cutaneous clinical form of leishmaniasis and were analyzed for proliferative responses to LACK antigen and SLA, frequency of lymphocyte subpopulations (analyzed ex vivo), and antigen-induced (LACK and SLA) cytokine production at the single-cell level (determined by flow cytometry). The following were determined. (i) The Th1-type response previously seen in patients with cutaneous leishmaniasis is due to gamma interferon (IFN-γ) production by several different sources, listed in order of contribution: CD4+ T lymphocytes, CD4−, CD8− lymphocytes, and CD8+ T lymphocytes. (ii) SLA induced a higher frequency of lymphocytes producing IFN-γ and tumor necrosis factor alpha (TNF-α) than did LACK. (iii) LACK induced an activation of monocyte populations as reflected by an increased percentage of CD14-positive cells. (iv) Neither SLA nor LACK induced detectable frequencies of cells producing interleukin-4 (IL-4) or IL-5. These data demonstrated a multifaceted immune response to SLA in human leishmaniasis involving Th1 CD4+ T lymphocytes (IFN-γ+ and IL-10−/IL-4−), Tc1 CD8+ T cells (IFN-γ+, and IL-10−/IL-4−), and a high frequency of TNF-α-producing lymphocytes. Moreover, it was determined that the recombinant antigen LACK acts as a weak inducer of Th1-type lymphocyte responses compared to SLA. PMID:11292745

  6. EPR-Spin Trapping and Flow Cytometric Studies of Free Radicals Generated Using Cold Atmospheric Argon Plasma and X-Ray Irradiation in Aqueous Solutions and Intracellular Milieu.

    Science.gov (United States)

    Uchiyama, Hidefumi; Zhao, Qing-Li; Hassan, Mariame Ali; Andocs, Gabor; Nojima, Nobuyuki; Takeda, Keigo; Ishikawa, Kenji; Hori, Masaru; Kondo, Takashi

    2015-01-01

    Electron paramagnetic resonance (EPR)-spin trapping and flow cytometry were used to identify free radicals generated using argon-cold atmospheric plasma (Ar-CAP) in aqueous solutions and intracellularly in comparison with those generated by X-irradiation. Ar-CAP was generated using a high-voltage power supply unit with low-frequency excitation. The characteristics of Ar-CAP were estimated by vacuum UV absorption and emission spectra measurements. Hydroxyl (·OH) radicals and hydrogen (H) atoms in aqueous solutions were identified with the spin traps 5,5-dimethyl-1-pyrroline N-oxide (DMPO), 3,3,5,5-tetramethyl-1-pyrroline-N-oxide (M4PO), and phenyl N-t-butylnitrone (PBN). The occurrence of Ar-CAP-induced pyrolysis was evaluated using the spin trap 3,5-dibromo-4-nitrosobenzene sulfonate (DBNBS) in aqueous solutions of DNA constituents, sodium acetate, and L-alanine. Human lymphoma U937 cells were used to study intracellular oxidative stress using five fluorescent probes with different affinities to a number of reactive species. The analysis and quantification of EPR spectra revealed the formation of enormous amounts of ·OH radicals using Ar-CAP compared with that by X-irradiation. Very small amounts of H atoms were detected whereas nitric oxide was not found. The formation of ·OH radicals depended on the type of rare gas used and the yield correlated inversely with ionization energy in the order of krypton > argon = neon > helium. No pyrolysis radicals were detected in aqueous solutions exposed to Ar-CAP. Intracellularly, ·OH, H2O2, which is the recombination product of ·OH, and OCl- were the most likely formed reactive oxygen species after exposure to Ar-CAP. Intracellularly, there was no practical evidence for the formation of NO whereas very small amounts of superoxides were formed. Despite the superiority of Ar-CAP in forming ·OH radicals, the exposure to X-rays proved more lethal. The mechanism of free radical formation in aqueous solutions and an

  7. EPR-Spin Trapping and Flow Cytometric Studies of Free Radicals Generated Using Cold Atmospheric Argon Plasma and X-Ray Irradiation in Aqueous Solutions and Intracellular Milieu.

    Directory of Open Access Journals (Sweden)

    Hidefumi Uchiyama

    Full Text Available Electron paramagnetic resonance (EPR-spin trapping and flow cytometry were used to identify free radicals generated using argon-cold atmospheric plasma (Ar-CAP in aqueous solutions and intracellularly in comparison with those generated by X-irradiation. Ar-CAP was generated using a high-voltage power supply unit with low-frequency excitation. The characteristics of Ar-CAP were estimated by vacuum UV absorption and emission spectra measurements. Hydroxyl (·OH radicals and hydrogen (H atoms in aqueous solutions were identified with the spin traps 5,5-dimethyl-1-pyrroline N-oxide (DMPO, 3,3,5,5-tetramethyl-1-pyrroline-N-oxide (M4PO, and phenyl N-t-butylnitrone (PBN. The occurrence of Ar-CAP-induced pyrolysis was evaluated using the spin trap 3,5-dibromo-4-nitrosobenzene sulfonate (DBNBS in aqueous solutions of DNA constituents, sodium acetate, and L-alanine. Human lymphoma U937 cells were used to study intracellular oxidative stress using five fluorescent probes with different affinities to a number of reactive species. The analysis and quantification of EPR spectra revealed the formation of enormous amounts of ·OH radicals using Ar-CAP compared with that by X-irradiation. Very small amounts of H atoms were detected whereas nitric oxide was not found. The formation of ·OH radicals depended on the type of rare gas used and the yield correlated inversely with ionization energy in the order of krypton > argon = neon > helium. No pyrolysis radicals were detected in aqueous solutions exposed to Ar-CAP. Intracellularly, ·OH, H2O2, which is the recombination product of ·OH, and OCl- were the most likely formed reactive oxygen species after exposure to Ar-CAP. Intracellularly, there was no practical evidence for the formation of NO whereas very small amounts of superoxides were formed. Despite the superiority of Ar-CAP in forming ·OH radicals, the exposure to X-rays proved more lethal. The mechanism of free radical formation in aqueous solutions and

  8. Global nuclear material flow/control model

    International Nuclear Information System (INIS)

    Dreicer, J.S.; Rutherford, D.S.; Fasel, P.K.; Riese, J.M.

    1997-01-01

    This is the final report of a two-year, Laboratory Directed Research and Development (LDRD) project at the Los Alamos National Laboratory (LANL). The nuclear danger can be reduced by a system for global management, protection, control, and accounting as part of an international regime for nuclear materials. The development of an international fissile material management and control regime requires conceptual research supported by an analytical and modeling tool which treats the nuclear fuel cycle as a complete system. The prototype model developed visually represents the fundamental data, information, and capabilities related to the nuclear fuel cycle in a framework supportive of national or an international perspective. This includes an assessment of the global distribution of military and civilian fissile material inventories, a representation of the proliferation pertinent physical processes, facility specific geographic identification, and the capability to estimate resource requirements for the management and control of nuclear material. The model establishes the foundation for evaluating the global production, disposition, and safeguards and security requirements for fissile nuclear material and supports the development of other pertinent algorithmic capabilities necessary to undertake further global nuclear material related studies

  9. Automatic coolant flow control device for a nuclear reactor assembly

    Science.gov (United States)

    Hutter, E.

    1984-01-27

    A device which controls coolant flow through a nuclear reactor assembly comprises a baffle means at the exit end of said assembly having a plurality of orifices, and a bimetallic member in operative relation to the baffle means such that at increased temperatures said bimetallic member deforms to unblock some of said orifices and allow increased coolant flow therethrough.

  10. Cell and nuclear enlargement of SW480 cells induced by a plant lignan, arctigenin: evaluation of cellular DNA content using fluorescence microscopy and flow cytometry.

    Science.gov (United States)

    Kang, Kyungsu; Lee, Hee Ju; Yoo, Ji-Hye; Jho, Eun Hye; Kim, Chul Young; Kim, Minkyun; Nho, Chu Won

    2011-08-01

    Arctigenin is a natural plant lignan previously shown to induce G(2)/M arrest in SW480 human colon cancer cells as well as AGS human gastric cancer cells, suggesting its use as a possible cancer chemopreventive agent. Changes in cell and nuclear size often correlate with the functionality of cancer-treating agents. Here, we report that arctigenin induces cell and nuclear enlargement of SW480 cells. Arctigenin clearly induced the formation of giant nuclear shapes in SW480, as demonstrated by fluorescence microscopic observation and quantitative determination of nuclear size. Cell and nuclear size were further assessed by flow cytometric analysis of light scattering and fluorescence pulse width after propidium iodide staining. FSC-H and FL2-W values (parameters referring to cell and nuclear size, respectively) significantly increased after arctigenin treatment; the mean values of FSC-H and FL2-W in arctigenin-treated SW480 cells were 572.6 and 275.1, respectively, whereas those of control cells were 482.0 and 220.7, respectively. Our approach may provide insights into the mechanism behind phytochemical-induced cell and nuclear enlargement as well as functional studies on cancer-treating agents.

  11. Heat transfer and fluid flow in nuclear systems

    CERN Document Server

    Fenech, Henri

    1982-01-01

    Heat Transfer and Fluid in Flow Nuclear Systems discusses topics that bridge the gap between the fundamental principles and the designed practices. The book is comprised of six chapters that cover analysis of the predicting thermal-hydraulics performance of large nuclear reactors and associated heat-exchangers or steam generators of various nuclear systems. Chapter 1 tackles the general considerations on thermal design and performance requirements of nuclear reactor cores. The second chapter deals with pressurized subcooled light water systems, and the third chapter covers boiling water reacto

  12. Nuclear-Coupled Flow Instabilities and Their Effects on Dryout

    Energy Technology Data Exchange (ETDEWEB)

    M. Ishii; X. Sunn; S. Kuran

    2004-09-27

    Nuclear-coupled flow/power oscillations in boiling water reactors (BWRs) are investigated experimentally and analytically. A detailed literature survey is performed to identify and classify instabilities in two-phase flow systems. The classification and the identification of the leading physical mechanisms of the two-phase flow instabilities are important to propose appropriate analytical models and scaling criteria for simulation. For the purpose of scaling and the analysis of the nonlinear aspects of the coupled flow/power oscillations, an extensive analytical modeling strategy is developed and used to derive both frequency and time domain analysis tools.

  13. On the elliptic flow for nearly symmetric collisions and nuclear ...

    Indian Academy of Sciences (India)

    The elliptic flow [10] has been proven to be one of the most fruitful probes for extracting the equation of state (EOS) and to ... at a very early stage of the nuclear collision, it is an excellent tool to probe the nuclear equation of state (EOS) under the ... The Plastic Ball group at the Bevalac in. Berkley were the first to quantify the ...

  14. Numerical simulation of interior flow field of nuclear model pump

    International Nuclear Information System (INIS)

    Wang Chunlin; Peng Na; Kang Can; Zhao Baitong; Zhang Hao

    2009-01-01

    Reynolds time-averaged N-S equations and the standard k-ε turbulent model were adopted, and three-dimensional non-structural of tetrahedral mesh division was used for modeling. Multiple reference frame model of rotating fluid mechanical model was used, under the design condition, the three-dimensional incompressible turbulent flow of nuclear model pump was simulated, and the results preferably post the characteristics of the interior flow field. This paper first analyzes the total pressure and velocity distribution in the flow field, and then describes the interior flow field characteristics of each part such as the impeller, diffuser and spherical shell, and also discusses the reasons that cause these characteristics. The study results can be used to estimate the performance of nuclear model pump, and will provide some useful references for its hydraulic optimized design. (authors)

  15. Cash flow forecasting model for nuclear power projects

    International Nuclear Information System (INIS)

    Liu Wei; Guo Jilin

    2002-01-01

    Cash flow forecasting is very important for owners and contractors of nuclear power projects to arrange the capital and to decrease the capital cost. The factors related to contractor cash flow forecasting are analyzed and a cash flow forecasting model is presented which is suitable for both contractors and owners. The model is efficiently solved using a cost-schedule data integration scheme described. A program is developed based on the model and verified with real project data. The result indicates that the model is efficient and effective

  16. A nuclear data acquisition system flow control model

    International Nuclear Information System (INIS)

    Hack, S.N.

    1988-01-01

    A general Petri Net representation of a nuclear data acquisition system model is presented. This model provides for the unique requirements of a nuclear data acquisition system including the capabilities of concurrently acquiring asynchronous and synchronous data, of providing multiple priority levels of flow control arbitration, and of permitting multiple input sources to reside at the same priority without the problem of channel lockout caused by a high rate data source. Finally, a previously implemented gamma camera/physiological signal data acquisition system is described using the models presented

  17. Initial angular momentum and flow in high energy nuclear collisions

    Science.gov (United States)

    Fries, Rainer J.; Chen, Guangyao; Somanathan, Sidharth

    2018-03-01

    We study the transfer of angular momentum in high energy nuclear collisions from the colliding nuclei to the region around midrapidity, using the classical approximation of the color glass condensate (CGC) picture. We find that the angular momentum shortly after the collision (up to times ˜1 /Qs , where Qs is the saturation scale) is carried by the "β -type" flow of the initial classical gluon field, introduced by some of us earlier. βi˜μ1∇iμ2-μ2∇iμ1 (i =1 ,2 ) describes the rapidity-odd transverse energy flow and emerges from Gauss's law for gluon fields. Here μ1 and μ2 are the averaged color charge fluctuation densities in the two nuclei, respectively. Interestingly, strong coupling calculations using anti-de Sitter/conformal field theory (AdS/CFT) techniques also find an energy flow term featuring this particular combination of nuclear densities. In classical CGC the order of magnitude of the initial angular momentum per rapidity in the reaction plane, at a time 1 /Qs , is |d L2/d η |≈ RAQs-3ɛ¯0/2 at midrapidity, where RA is the nuclear radius, and ɛ¯0 is the average initial energy density. This result emerges as a cancellation between a vortex of energy flow in the reaction plane aligned with the total angular momentum, and energy shear flow opposed to it. We discuss in detail the process of matching classical Yang-Mills results to fluid dynamics. We will argue that dissipative corrections should not be discarded to ensure that macroscopic conservation laws, e.g., for angular momentum, hold. Viscous fluid dynamics tends to dissipate the shear flow contribution that carries angular momentum in boost-invariant fluid systems. This leads to small residual angular momentum around midrapidity at late times for collisions at high energies.

  18. Effects of Gogny type interactions on the nuclear flow

    International Nuclear Information System (INIS)

    Sebille, F.; De La Mota, V.; Jouault, B.; Schuck, P.

    1995-01-01

    A flow analysis on symmetric and asymmetric reactions from 100 to 400 MeV/n is performed in the framework of the semi-classical Landau-Vlasov approach. In this energy range our results present two different trends. At lower energies it is governed by the momentum dependence of the nuclear optical potential, whereas at higher energies its density dependence plays a crucial role leading to a rather pronounced sensitivity of the incompressibility modulus. The non-locality of the nuclear interaction is relevant for asymmetric colliding systems. With an incompressibility modulus in the vicinity of 200 MeV, an excellent quantitative description of the flow behavior with incident energy and impact parameter or the system mass is provided. (authors)., 26 refs., 6 figs., 3 tabs

  19. Plant DNA flow cytometry and estimation of nuclear genome size

    Czech Academy of Sciences Publication Activity Database

    Doležel, Jaroslav; Bartoš, Jan

    2005-01-01

    Roč. 95, - (2005), s. 99-110 ISSN 0305-7364 R&D Projects: GA ČR GA522/03/0354; GA ČR GA204/04/1207 Institutional research plan: CEZ:AV0Z50380511 Keywords : flow cytometry * nuclear genome size * DNA C-value Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.665, year: 2005

  20. Modeling of Flow in Nuclear Reactor Fuel Cell Outlet

    Directory of Open Access Journals (Sweden)

    František URBAN

    2010-12-01

    Full Text Available Safe and effective load of nuclear reactor fuel cells demands qualitative and quantitative analysis of relations between coolant temperature in fuel cell outlet temperature measured by thermocouple and middle temperature of coolant in thermocouple plane position. In laboratory at Insitute of thermal power engineering of the Slovak University of Technology in Bratislava was installed an experimental physical fuel cell model of VVER 440 nuclear power plant with V 213 nuclear reactors. Objective of measurements on physical model was temperature and velocity profiles analysis in the fuel cell outlet. In this paper the measured temperature and velocity profiles are compared with the results of CFD simulation of fuel cell physical model coolant flow.

  1. Effective flow-accelerated corrosion programs in nuclear facilities

    International Nuclear Information System (INIS)

    Esselman, Thomas C.; McBrine, William J.

    2004-01-01

    Piping Flow-Accelerated Corrosion Programs in nuclear power generation facilities are classically comprised of the selection of inspection locations with the assistance of a predictive methodology such as the Electric Power Research Institute computer codes CHECMATE or CHECWORKS, performing inspections, conducting structural evaluations on the inspected components, and implementing the appropriate sample expansion and corrective actions. Performing such a sequence of steps can be effective in identifying thinned components and implementing appropriate short term and long term actions necessary to resolve flow-accelerated corrosion related problems. A maximally effective flow-accelerated corrosion (FAC) program requires an understanding of many programmatic details. These include the procedural control of the program, effective use of historical information, managing the activities performed during a limited duration outage, allocating resources based on risk allocation, having an acute awareness of how the plant is operated, investigating components removed from the plant, and several others. This paper will describe such details and methods that will lead to a flow-accelerated corrosion program that effectively minimizes the risk of failure due to flow-accelerated corrosion and provide full and complete documentation of the program. (author)

  2. Review of coaxial flow gas core nuclear rocket fluid mechanics

    International Nuclear Information System (INIS)

    Weinstein, H.

    1976-01-01

    In a prematurely aborted attempt to demonstrate the feasibility of using a gas core nuclear reactor as a rocket engine, NASA initiated a number of studies on the relevant fluid mechanics problems. These studies were carried out at NASA laboratories, universities and industrial research laboratories. Because of the relatively sudden termination of most of this work, a unified overview was never presented which demonstrated the accomplishments of the program and pointed out the areas where additional work was required for a full understanding of the cavity flow. This review attempts to fulfill a part of this need in two important areas

  3. Multilevel Flow Modeling for Nuclear Power Plant Diagnosis

    DEFF Research Database (Denmark)

    Gola, G; Lind, Morten; Thunem, Harald P-J

    2012-01-01

    As complexity and safety requirements of current and future nuclear power plants increase, innovative methods are being investigated to perform accurate and reliable system diagnoses. Detecting malfunctions, identifying their causes and possibly predicting their consequences are major challenges......-scale monitoring systems is hard to handle manually. In this paper, the use of an innovative function-oriented modeling approach, called Multilevel Flow Modeling, is proposed for performing an automatic analysis of the outcomes of the monitoring systems with the aim of identifying the root causes of the possibly...

  4. Estimation of nuclear DNA content in plants using flow cytometry

    Czech Academy of Sciences Publication Activity Database

    Doležel, Jaroslav; Greilhuber, J.; Suda, Jan

    2007-01-01

    Roč. 2, č. 9 (2007), s. 2233-2244 ISSN 1754-2189 R&D Projects: GA MŠk ME 884; GA MŠk(CZ) LC06004 Institutional research plan: CEZ:AV0Z50380511; CEZ:AV0Z60050516 Source of funding: V - iné verejné zdroje ; V - iné verejné zdroje Keywords : Flow cytometry * nuclear DNA content * suspensions of intact nuclei Subject RIV: EB - Genetics ; Molecular Biology www.nature.com/nprot/journal/v2/n9/abs/nprot.2007.310.html

  5. Multilevel flow modeling of Monju Nuclear Power Plant

    DEFF Research Database (Denmark)

    Lind, Morten; Yoshikawa, Hidekazu; Jørgensen, Sten Bay

    2011-01-01

    functions and structure. The paper will describe how MFM can be used to represent the goals and functions of the Japanese Monju Nuclear Power Plant. A detailed explanation will be given of the model describing the relations between levels of goal, function and structural. Furthermore, it will be explained......Multilevel Flow Modeling is a method for modeling complex processes on multiple levels of means-end and part-whole abstraction. The modeling method has been applied on a wide range of processes including power plants, chemical engineering plants and power systems. The modeling method is supported...

  6. Progress in the Development of Compressible, Multiphase Flow Modeling Capability for Nuclear Reactor Flow Applications

    Energy Technology Data Exchange (ETDEWEB)

    R. A. Berry; R. Saurel; F. Petitpas; E. Daniel; O. Le Metayer; S. Gavrilyuk; N. Dovetta

    2008-10-01

    In nuclear reactor safety and optimization there are key issues that rely on in-depth understanding of basic two-phase flow phenomena with heat and mass transfer. Within the context of multiphase flows, two bubble-dynamic phenomena – boiling (heterogeneous) and flashing or cavitation (homogeneous boiling), with bubble collapse, are technologically very important to nuclear reactor systems. The main difference between boiling and flashing is that bubble growth (and collapse) in boiling is inhibited by limitations on the heat transfer at the interface, whereas bubble growth (and collapse) in flashing is limited primarily by inertial effects in the surrounding liquid. The flashing process tends to be far more explosive (and implosive), and is more violent and damaging (at least in the near term) than the bubble dynamics of boiling. However, other problematic phenomena, such as crud deposition, appear to be intimately connecting with the boiling process. In reality, these two processes share many details.

  7. Flow accelerated corrosion in Indian nuclear power plants

    International Nuclear Information System (INIS)

    Roychowdhury, S.; Ahmedabadi, P.; Kain, V.; Barua, D.K.

    2008-01-01

    Flow accelerated corrosion (FAC) related failures have occurred in a large number of pipelines in the secondary as well as primary section of light water reactors worldwide. This is primarily a corrosion process enhanced by (electro) chemical dissolution and mass transfer, rather than a mechanical process. FAC is an extension of the generalized carbon steel corrosion process in stagnant water. The mechanism of FAC has been described in this paper. It has caused rupture of large, medium and small diameter pipelines carrying either single phase water or two phase (wet steam) flow. It is reported to be the only mechanism that has significant potential for large leaks in the secondary circuit. This paper describes the various FAC related incidents that have occurred in the secondary as well as primary side of Indian Pressurized Heavy Water Reactors. Failure analysis done on the 10% feed water line in KAPS, NRV in auxiliary feed water system of KAPS, and affected pipeline components from the secondary water system of KGS has been described in this paper. The surface features observed (the signature of FAC) on the ID of the pipelines affected by FAC in the primary and the secondary circuits of nuclear reactors have been described. Signatures of FAC, both single phase FAC and dual phase FAC related failures and the signature pattern formed on the surfaces in both the primary and the secondary circuit of the Indian nuclear power plants have been described in this paper. (author)

  8. Computer simulation of two-phase flow in nuclear reactors

    International Nuclear Information System (INIS)

    Wulff, W.

    1993-01-01

    Two-phase flow models dominate the requirements of economic resources for the development and use of computer codes which serve to analyze thermohydraulic transients in nuclear power plants. An attempt is made to reduce the effort of analyzing reactor transients by combining purpose-oriented modelling with advanced computing techniques. Six principles are presented on mathematical modeling and the selection of numerical methods, along with suggestions on programming and machine selection, all aimed at reducing the cost of analysis. Computer simulation is contrasted with traditional computer calculation. The advantages of run-time interactive access operation in a simulation environment are demonstrated. It is explained that the drift-flux model is better suited than the two-fluid model for the analysis of two-phase flow in nuclear reactors, because of the latter's closure problems. The advantage of analytical over numerical integration is demonstrated. Modeling and programming techniques are presented which minimize the number of needed arithmetical and logical operations and thereby increase the simulation speed, while decreasing the cost. (orig.)

  9. Turbulence coefficients and stability studies for the coaxial flow or dissimiliar fluids. [gaseous core nuclear reactors

    Science.gov (United States)

    Weinstein, H.; Lavan, Z.

    1975-01-01

    Analytical investigations of fluid dynamics problems of relevance to the gaseous core nuclear reactor program are presented. The vortex type flow which appears in the nuclear light bulb concept is analyzed along with the fluid flow in the fuel inlet region for the coaxial flow gaseous core nuclear reactor concept. The development of numerical methods for the solution of the Navier-Stokes equations for appropriate geometries is extended to the case of rotating flows and almost completes the gas core program requirements in this area. The investigations demonstrate that the conceptual design of the coaxial flow reactor needs further development.

  10. Modelling the work flow of a nuclear waste management program

    International Nuclear Information System (INIS)

    Hoeyer Mortensen, K.; Pinci, V.

    1997-03-01

    In this paper we describe a modelling project to improve a nuclear waste management program in charge of the creation of a new system for the permanent disposal of nuclear waste. SADT (Structural Analysis and Design Technique) is used in order to provide a work-flow description of the functions to be performed by the waste management program. This description is then translated into a number of Coloured Petri Nets (CPN or CP-nets) corresponding to different program functions where additional behavioural inscriptions provide basis for simulation. Each of these CP-nets is simulated to produce timed event charts that are useful for understanding the behaviour of the program functions under different scenarios. Then all the CPN models are linked together to form a single stand-alone application that is useful for validating the interaction and cooperation between the different program functions. A technique for linking executable CPN models is developed for supporting large modelling projects and parallel development of independent CPN models. (au) 11 refs

  11. Neural network modeling of chaotic dynamics in nuclear reactor flows

    International Nuclear Information System (INIS)

    Welstead, S.T.

    1992-01-01

    Neural networks have many scientific applications in areas such as pattern classification and time series prediction. The universal approximation property of these networks, however, can also be exploited to provide researchers with tool for modeling observed nonlinear phenomena. It has been shown that multilayer feed forward networks can capture important global nonlinear properties, such as chaotic dynamics, merely by training the network on a finite set of observed data. The network itself then provides a model of the process that generated the data. Characterizations such as the existence and general shape of a strange attractor and the sign of the largest Lyapunov exponent can then be extracted from the neural network model. In this paper, the author applies this idea to data generated from a nonlinear process that is representative of convective flows that can arise in nuclear reactor applications. Such flows play a role in forced convection heat removal from pressurized water reactors and boiling water reactors, and decay heat removal from liquid-metal-cooled reactors, either by natural convection or by thermosyphons

  12. A Triple-Stain Flow Cytometric Method to Assess Plasma- and Acrosome-Membrane Integrity of Cryopreserved Bovine Sperm Immediately after Thawing in Presence of Egg-Yolk Particles

    NARCIS (Netherlands)

    Nagy, S.; Jansen, J.; Topper, E.K.; Gadella, B.M.

    2003-01-01

    Simultaneously evaluating postthaw viability and acrosome integrity of spermatozoa by flow cytometry would provide a valuable testing tool in both research and routine work. In the present study, a new triple-stain combination was developed for the simultaneous evaluation of viability and acrosome

  13. Nuclear Technology Series. Course 4: Heat Transfer and Fluid Flow.

    Science.gov (United States)

    Center for Occupational Research and Development, Inc., Waco, TX.

    This technical specialty course is one of thirty-five courses designed for use by two-year postsecondary institutions in five nuclear technician curriculum areas: (1) radiation protection technician, (2) nuclear instrumentation and control technician, (3) nuclear materials processing technician, (4) nuclear quality-assurance/quality-control…

  14. Two-phase flow heat transfer in nuclear reactor systems

    International Nuclear Information System (INIS)

    Koncar, Bostjan; Krepper, Eckhard; Bestion, Dominique; Song, Chul-Hwa; Hassan, Yassin A.

    2013-01-01

    Complete text of publication follows: Heat transfer and phase change phenomena in two-phase flows are often encountered in nuclear reactor systems and are therefore of paramount importance for their optimal design and safe operation.The complex phenomena observed especially during transient operation of nuclear reactor systems necessitate extensive theoretical and experimental investigations. This special issue brings seven research articles of high quality. Though small in number, they cover a wide range of topics, presenting high complexity and diversity of heat transfer phenomena in two-phase flow. In the last decades a vast amount of research has been devoted to theoretical work and computational simulations, yet the experimental work remains indispensable for understanding of two-phase flow phenomena and for model validation purposes. This is reflected also in this issue, where only one article is purely experimental, while three of them deal with theoretical modelling and the remaining three with numerical simulations. The experimental investigation of the critical heat flux (CHF) phenomena by means of photographic study is presented in the paper of J. Park et al. They have used a high-speed camera system to observe the transient boiling characteristics on a thin horizontal cylinder submerged in a pool of water or highly wetting liquid. Experiments show that the initial boiling process is strongly affected by the properties and wettability of the liquid. The authors have stressed the importance of the local scale observation leading to better understanding of the transient CHF phenomena. In the article of G. Espinosa-Paredes et al. a theoretical work concerning the derivation of transport equations for two-phase flow is presented. The author proposes a novel approach based on derivation of nonlocal volume averaged equations which contain new terms related to nonlocal transport effects. These non-local terms act as coupling elements between the phenomena

  15. Measurement of two phase flow properties using the nuclear reactor instruments

    International Nuclear Information System (INIS)

    Albrecht, R.W.; Washington Univ., Seattle; Crowe, R.D.; Dailey, D.J.; Kosaly, G.; Damborg, M.J.

    1982-01-01

    A procedure is introduced for characterizing one dimensional, two phase flow in terms of three properties; propagation, structure, and dynamics. It is shown that all of these properties can be measured by analyzing the response of the reactor neutron field to a two phase flow perturbation. Therefore, a nuclear reactor can be regarded as a two phase flow instrument. (author)

  16. Intestinal intraepithelial lymphocyte cytometric pattern is more accurate than subepithelial deposits of anti-tissue transglutaminase IgA for the diagnosis of celiac disease in lymphocytic enteritis.

    Directory of Open Access Journals (Sweden)

    Fernando Fernández-Bañares

    Full Text Available BACKGROUND & AIMS: An increase in CD3+TCRγδ+ and a decrease in CD3- intraepithelial lymphocytes (IEL is a characteristic flow cytometric pattern of celiac disease (CD with atrophy. The aim was to evaluate the usefulness of both CD IEL cytometric pattern and anti-TG2 IgA subepithelial deposit analysis (CD IF pattern for diagnosing lymphocytic enteritis due to CD. METHODS: Two-hundred and five patients (144 females who underwent duodenal biopsy for clinical suspicion of CD and positive celiac genetics were prospectively included. Fifty had villous atrophy, 70 lymphocytic enteritis, and 85 normal histology. Eight patients with non-celiac atrophy and 15 with lymphocytic enteritis secondary to Helicobacter pylori acted as control group. Duodenal biopsies were obtained to assess both CD IEL flow cytometric (complete or incomplete and IF patterns. RESULTS: Sensitivity of IF, and complete and incomplete cytometric patterns for CD diagnosis in patients with positive serology (Marsh 1+3 was 92%, 85 and 97% respectively, but only the complete cytometric pattern had 100% specificity. Twelve seropositive and 8 seronegative Marsh 1 patients had a CD diagnosis at inclusion or after gluten free-diet, respectively. CD cytometric pattern showed a better diagnostic performance than both IF pattern and serology for CD diagnosis in lymphocytic enteritis at baseline (95% vs 60% vs 60%, p = 0.039. CONCLUSIONS: Analysis of the IEL flow cytometric pattern is a fast, accurate method for identifying CD in the initial diagnostic biopsy of patients presenting with lymphocytic enteritis, even in seronegative patients, and seems to be better than anti-TG2 intestinal deposits.

  17. Flowing gas, non-nuclear experiments on the gas core reactor

    Science.gov (United States)

    Kunze, J. F.; Cooper, C. G.; Macbeth, P. J.

    1973-01-01

    Variations in cavity wall and injection configurations of the gas core reactor were aimed at establishing flow patterns that give a maximum of the nuclear criticality eigenvalue. Correlation with the nuclear effect was made using multigroup diffusion theory normalized by previous benchmark critical experiments. Air was used to simulate the hydrogen propellant in the flow tests, and smoked air, argon, or Freon to simulate the central nuclear fuel gas. Tests were run both in the down-firing and upfiring directions. Results showed that acceptable flow patterns with volume fraction for the simulated nuclear fuel gas and high flow rate ratios of propellant to fuel can be obtained. Using a point injector for the fuel, good flow patterns are obtained by directing the outer gas at high velocity long the cavity wall, using louvered injection schemes. Recirculation patterns were needed to stabilize the heavy central gas when different gases are used.

  18. Flow cytometric analysis of mitotic cycle perturbation by chemical carcinogens in cultured epithelial cells. [Effects of benzo(a)pyrene-diol-epoxide on mitotic cycle of cultural mouse liver epithelial cells

    Energy Technology Data Exchange (ETDEWEB)

    Pearlman, Andrew Leonard [Univ. of California, Berkeley, CA (United States)

    1978-08-01

    A system for kinetic analysis of mitotic cycle perturbation by various agents was developed and applied to the study of the mitotic cycle effects and dependency of the chemical carcinogen benzo(a)pyrene-diolepoxide, DE, upon a mouse lever epithelial cell line, NMuLi. The study suggests that the targets of DE action are not confined to DNA alone but may include cytoplasmic structures as well. DE was found to affect cells located in virtually every phase of the mitotic cycle, with cells that were actively synthesizing DNA showing the strongest response. However, the resulting perturbations were not confined to S-phase alone. DE slowed traversal through S-phase by about 40% regardless of the cycle phase of the cells exposed to it, and slowed traversal through G2M by about 50%. When added to G1 cells, DE delayed recruitment of apparently quiescent (G0) cells by 2 hours, and reduced the synchrony of the cohort of cells recruited into active proliferation. The kinetic analysis system consists of four elements: tissue culture methods for propagating and harvesting cell populations; an elutriation centrifugation system for bulk synchronization of cells in various phases of the mitotic cycle; a flow cytometer (FCM), coupled with appropriate staining protocols, to enable rapid analysis of the DNA distribution of any given cell population; and data reduction and analysis methods for extracting information from the DNA histograms produced by the FCM. The elements of the system are discussed. A mathematical analysis of DNA histograms obtained by FCM is presented. The analysis leads to the detailed implementation of a new modeling approach. The new modeling approach is applied to the estimation of cell cycle kinetic parameters from time series of DNA histograms, and methods for the reduction and interpretation of such series are suggested.

  19. A review of flow analysis methods for determination of radionuclides in nuclear wastes and nuclear reactor coolants.

    Science.gov (United States)

    Trojanowicz, Marek; Kołacińska, Kamila; Grate, Jay W

    2018-06-01

    The safety and security of nuclear power plant operations depend on the application of the most appropriate techniques and methods of chemical analysis, where modern flow analysis methods prevail. Nevertheless, the current status of the development of these methods is more limited than it might be expected based on their genuine advantages. The main aim of this paper is to review the automated flow analysis procedures developed with various detection methods for the nuclear energy industry. The flow analysis methods for the determination of radionuclides, that have been reported to date, are primarily focused on their environmental applications. The benefits of the application of flow methods in both monitoring of the nuclear wastes and process analysis of the primary circuit coolants of light water nuclear reactors will also be discussed. The application of either continuous flow methods (CFA) or injection methods (FIA, SIA) of the flow analysis with the β-radiometric detection shortens the analysis time and improves the precision of determination due to mechanization of certain time-consuming operations of the sample processing. Compared to the radiometric detection, the mass spectrometry (MS) detection enables one to perform multicomponent analyses as well as the determination of transuranic isotopes with much better limits of detection. Copyright © 2018 Elsevier B.V. All rights reserved.

  20. Seasonality in molecular and cytometric diversity of marine bacterioplankton: the reshuffling of bacterial taxa by vertical mixing

    KAUST Repository

    García, Francisca C.

    2015-07-17

    The ’cytometric diversity’ of phytoplankton communities has been studied based on single-cell properties, but the applicability of this method to characterize bacterioplankton has been unexplored. Here, we analysed seasonal changes in cytometric diversity of marine bacterioplankton along a decadal time-series at three coastal stations in the Southern Bay of Biscay. Shannon-Weaver diversity estimates and Bray-Curtis similarities obtained by cytometric and molecular (16S rRNA tag sequencing) methods were significantly correlated in samples from a 3.5-year monthly time-series. Both methods showed a consistent cyclical pattern in the diversity of surface bacterial communities with maximal values in winter. The analysis of the highly resolved flow cytometry time-series across the vertical profile showed that water column mixing was a key factor explaining the seasonal changes in bacterial composition and the winter increase in bacterial diversity in coastal surface waters. Due to its low cost and short processing time as compared to genetic methods, the cytometric diversity approach represents a useful complementary tool in the macroecology of aquatic microbes.

  1. Global flow of glasma in high energy nuclear collisions

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Guangyao; Fries, Rainer J., E-mail: rjfries@comp.tamu.edu

    2013-06-25

    We discuss the energy flow of the classical gluon fields created in collisions of heavy nuclei at collider energies. We show how the Yang–Mills analog of Faraday's Law and Gauss' Law predicts the initial gluon flux tubes to expand or bend. The resulting transverse and longitudinal structure of the Poynting vector field has a rich phenomenology. Besides the well-known radial and elliptic flow in transverse direction, classical quantum chromodynamics predicts a rapidity-odd transverse flow that tilts the fireball for non-central collisions, and it implies a characteristic flow pattern for collisions of non-symmetric systems A+B. The rapidity-odd transverse flow translates into a directed particle flow v{sub 1} which has been observed at RHIC and LHC. The global flow fields in heavy ion collisions could be a powerful check for the validity of classical Yang–Mills dynamics in high energy collisions.

  2. Study of visualized simulation and analysis of nuclear fuel cycle system based on multilevel flow model

    International Nuclear Information System (INIS)

    Liu Jingquan; Yoshikawa, H.; Zhou Yangping

    2005-01-01

    Complex energy and environment system, especially nuclear fuel cycle system recently raised social concerns about the issues of economic competitiveness, environmental effect and nuclear proliferation. Only under the condition that those conflicting issues are gotten a consensus between stakeholders with different knowledge background, can nuclear power industry be continuingly developed. In this paper, a new analysis platform has been developed to help stakeholders to recognize and analyze various socio-technical issues in the nuclear fuel cycle sys- tem based on the functional modeling method named Multilevel Flow Models (MFM) according to the cognition theory of human being, Its character is that MFM models define a set of mass, energy and information flow structures on multiple levels of abstraction to describe the functional structure of a process system and its graphical symbol representation and the means-end and part-whole hierarchical flow structure to make the represented process easy to be understood. Based upon this methodology, a micro-process and a macro-process of nuclear fuel cycle system were selected to be simulated and some analysis processes such as economics analysis, environmental analysis and energy balance analysis related to those flows were also integrated to help stakeholders to understand the process of decision-making with the introduction of some new functions for the improved Multilevel Flow Models Studio, and finally the simple simulation such as spent fuel management process simulation and money flow of nuclear fuel cycle and its levelised cost analysis will be represented as feasible examples. (authors)

  3. Flow Cytometric Applicability of Fluorescent Vitality Probes on Phytoplankton

    NARCIS (Netherlands)

    Peperzak, L.; Brussaard, C.P.D.

    2011-01-01

    The applicability of six fluorescent probes (four esterase probes: acetoxymethyl ester of Calcein [Calcein-AM], 5-chloromethylfluorescein diacetate [CMFDA], fluorescein diacetate [FDA], and 2',7'-dichlorofluorescein diacetate [H(2)DCFDA]; and two membrane probes: bis-(1,3-dibutylbarbituric acid)

  4. FLOW CYTOMETRIC APPLICABILITY OF FLUORESCENT VITALITY PROBES ON PHYTOPLANKTON1.

    Science.gov (United States)

    Peperzak, Louis; Brussaard, Corina P D

    2011-06-01

    The applicability of six fluorescent probes (four esterase probes: acetoxymethyl ester of Calcein [Calcein-AM], 5-chloromethylfluorescein diacetate [CMFDA], fluorescein diacetate [FDA], and 2',7'-dichlorofluorescein diacetate [H 2 DCFDA]; and two membrane probes: bis-(1,3-dibutylbarbituric acid) trimethine oxonol [DiBAC 4 (3)] and SYTOX-Green) as vitality stains was tested on live and killed cells of 40 phytoplankton strains in exponential and stationary growth phases, belonging to 12 classes and consisting of four cold-water, 26 temperate, and four warm-water species. The combined live/dead ratios of all six probes indicated significant differences between the 12 plankton classes (P live/dead ratios of FDA and CMFDA were not significantly different from each other, and both performed better than Calcein-AM and H 2 DCFDA (P live/dead ratios) among all six probes belonged to nine genera from six classes of phytoplankton. In conclusion, FDA, CMFDA, DIBAC 4 (3), and SYTOX-Green represent a wide choice of vitality probes in the study of phytoplankton ecology, applicable in many species from different algal classes, originating from different regions and at different stages of growth. © 2011 Phycological Society of America.

  5. Flow Cytometric Ploidy Determination of Oral Premalignant and Malignant Lesions

    Science.gov (United States)

    1990-01-01

    identifiable lesions, some were associated with oral white lesions, either concomitant or precedent. These white lesions were referred to as leukoplakia , a...widely variable results. These authors undertook a clinical-pathologic correlation of oral leukoplakia encountered in the biopsy services of two...with oral leukoplakia for an average period of 7.2 years. All lesions were more than one centimeter in size and had been present and observed for a

  6. Flow cytometric quantification of radiation responses of murine peritoneal cells

    International Nuclear Information System (INIS)

    Tokita, N.; Raju, M.R.

    1982-01-01

    Methods have been developed to distinguish subpopulations of murine peritoneal cells, and these were applied to the measurement of early changes in peritoneal cells after irradiation. The ratio of the two major subpopulations in the peritoneal fluid, lymphocytes and macrophages, was measured rapidly by means of cell volume distribution analysis as well as by hypotonic propidium iodide (PI) staining. After irradiation, dose and time dependent changes were noted in the cell volume distributions: a rapid loss of peritoneal lymphocytes, and an increase in the mean cell volume of macrophages. The hypotonic PI staining characteristics of the peritoneal cells showed two or three distinctive G 1 peaks. The ratio of the areas of these peaks was also found to be dependent of the radiation dose and the time after irradiation. These results demonstrate that these two parameters may be used to monitor changes induced by irradiation (biological dosimetry), and to sort different peritoneal subpopulations

  7. Flow cytometric analysis of bone marrow leukocytes in neonatal dogs

    Czech Academy of Sciences Publication Activity Database

    Faldyna, M.; Šinkora, Jiří; Knotigová, P.; Řeháková, Zuzana; Morávková, Alena; Toman, M.

    2003-01-01

    Roč. 95, - (2003), s. 165-176 ISSN 0165-2427 R&D Projects: GA ČR GA524/00/0474; GA ČR GP524/02/P010 Institutional research plan: CEZ:AV0Z5020903 Keywords : cd34 * sirp * b cell Subject RIV: EC - Immunology Impact factor: 1.652, year: 2003

  8. Howard University Flow Cytometric Sorter For Research and Education

    Science.gov (United States)

    2015-08-04

    based on beta -D- galactosidase activity after transduction of Escherichia coli lacZ. Proceedings of the National Academy of Sciences of the United... beta -D- galactosidase activity after transduction of Escherichia coli lacZ. Proceedings of the National Academy of Sciences of the United States of

  9. Rapid Flow cytometric prenatal diagnosis of primary immunodeficiency (PID) disorders.

    Science.gov (United States)

    Mishra, Anju; Gupta, Maya; Dalvi, Aparna; Ghosh, Kanjaksha; Madkaikar, Manisha

    2014-04-01

    Primary Immunodeficiency diseases (PID) are a heterogeneous group of inherited disorders of immune system. Immunophenotypic evaluation of PIDs using flowcytometry provides important clues for diagnosis of these disorders, though confirmation requires identification of underlying molecular defects. Prenatal diagnosis (PND) forms an important component of management in families affected with severe PID. However, molecular diagnostic facilities for each of these diseases are not available and may not be possible to perform in all cases. In such scenario we opted for phenotypic prenatal diagnosis by cordocentesis for families with index case having immunophenotypically well characterized PID. Normal reference ranges of lymphocyte subsets, CD 18/CD11 integrins on leukocytes, MHC class II expression and oxidative burst activity of fetal neutrophils at 18 weeks of gestation were previously established on 30 cord blood samples. PND was performed in 13 families with PIDs. Maternal contamination was ruled out by VNTR analysis. Out of 13 fetuses, nine were found to be unaffected (three cases with leukocyte adhesion deficiency (LAD-I), four cases with severe combined immunodeficiency diseases (SCID), one with X-linked agammaglobulinemia (XLA), and one with chronic granulomatous disease (CGD)] and three were found to be affected (one with T-B+NK-SCID, one with MHC class II deficiency and one with LAD-I). Diagnosis was confirmed by testing the cord blood samples after delivery and further follow-up of the children. In one family diagnosis could not be offered due to maternal contamination. No procedure related complications were observed. Flowcytometry offers rapid and sensitive method for prenatal diagnosis and genetic counseling for selected phenotypically well characterized PID in cases where molecular diagnostic facilities are not available.

  10. Flow cytometric detection of viruses in the Zuari estuary, Goa

    Digital Repository Service at National Institute of Oceanography (India)

    Mitbavkar, S.; Rajaneesh, K.M.; SathishKumar, P.

    abundance 3 . Viruses are the smallest known organisms (20–200 nm) with simple biological structures consist- ing of nucleic acid, either DNA or RNA (single- or double-stranded), with a pro- tein coat (capsid). Viruses are known to infect prokaryo- tes... in the microbial loop. Because prokaryotes and autotrophic and heterotrophic protists play pivotal roles in the biogeochemical cycles and global ocean functioning, viral infections of these groups of organ- ism have important ecological conse- quences 2...

  11. Recycling flow control device for a nuclear reactor

    International Nuclear Information System (INIS)

    Shida, Toichi; Nakamura, Hideo; Hirose, Masao; Akiyama, Takao.

    1975-01-01

    Object: To permit a valve operation test to be periodically made during plant operation without causing variations in plant power by detecting flow control valve defect on the basis of a valve aperture alteration instruction. Structure: Step signals which are equal in absolute value and opposite in sign are coupled to the input side of flow controllers provided on the recycling loops of two or more recycling flow control systems. With these inputs the aperture of the flow control valve on one side is increased (or reduced) while the aperture of the valve on the other side is reduced (or increased). As a result, the recycling flow rate in the loop on one side is increased (or reduced) while that on the other side is reduced (or increased). Whether the valve is normally operating or not is confirmed by checking the recycling flow rate and valve aperture. (Nakamura, S.)

  12. On-line validation of feedwater flow rate in nuclear power plants using neural networks

    International Nuclear Information System (INIS)

    Khadem, M.; Ipakchi, A.; Alexandro, F.J.; Colley, R.W.

    1994-01-01

    On-line calibration of feedwater flow rate measurement in nuclear power plants provides a continuous realistic value of feedwater flow rate. It also reduces the manpower required for periodic calibration needed due to the fouling and defouling of the venturi meter surface condition. This paper presents a method for on-line validation of feedwater flow rate in nuclear power plants. The method is an improvement of the previously developed method which is based on the use of a set of process variables dynamically related to the feedwater flow rate. The online measurements of this set of variables are used as inputs to a neural network to obtain an estimate of the feedwater flow rate reading. The difference between the on-line feedwater flow rate reading, and the neural network estimate establishes whether there is a need to apply a correction factor to the feedwater flow rate measurement for calculation of the actual reactor power. The method was applied to the feedwater flow meters in the two feedwater flow loops of the TMI-1 nuclear power plant. The venturi meters used for flow measurements are susceptible to frequent fouling that degrades their measurement accuracy. The fouling effects can cause an inaccuracy of up to 3% relative error in feedwater flow rate reading. A neural network, whose inputs were the readings of a set of reference instruments, was designed to predict both feedwater flow rates simultaneously. A multi-layer feedforward neural network employing the backpropagation algorithm was used. A number of neural network training tests were performed to obtain an optimum filtering technique of the input/output data of the neural networks. The result of the selection of the filtering technique was confirmed by numerous Fast Fourier Transform (FFT) tests. Training and testing were done on data from TMI-1 nuclear power plant. The results show that the neural network can predict the correct flow rates with an absolute relative error of less than 2%

  13. Cytometric Approach for Detection of Encephalitozoon intestinalis, an Emergent Agent▿

    Science.gov (United States)

    Barbosa, Joana; Rodrigues, Acácio Gonçalves; Pina-Vaz, Cidália

    2009-01-01

    Encephalitozoon intestinalis is responsible for intestinal disease in patients with AIDS and immunocompetent patients. The infectious form is a small spore that is resistant to water treatment procedures. Its detection is very important, but detection is very cumbersome and time-consuming. Our main objective was to develop and optimize a specific flow cytometric (FC) protocol for the detection of E. intestinalis in hospital tap water and human feces. To determine the optimal specific antibody (Microspor-FA) concentration, a known concentration of E. intestinalis spores (Waterborne, Inc.) was suspended in hospital tap water and stool specimens with different concentrations of Microspor-FA, and the tap water and stool specimens were incubated under different conditions. The sensitivity limit and specificity were also evaluated. To study spore infectivity, double staining with propidium iodide (PI) and Microspor-FA was undertaken. Distinct approaches for filtration and centrifugation of the stool specimens were used. E. intestinalis spores stained with 10 μg/ml of Microspor-FA at 25°C overnight provided the best results. The detection limit was 5 × 104 spores/ml, and good specificity was demonstrated. Simultaneous staining with Microspor-FA and PI ensured that the E. intestinalis spores were dead and therefore noninfectious. With the stool specimens, better spore recovery was observed with a saturated solution of NaCl and centrifugation at 1,500 × g for 15 min. A new approach for the detection of E. intestinalis from tap water or human feces that ensures that the spores are not viable is now available and represents an important step for the prevention of this threat to public health. PMID:19439525

  14. Survey of near-field flow calculations for nuclear waste repositories NMA L21

    International Nuclear Information System (INIS)

    Wilson, R.N.; Holland, D.H.

    1979-11-01

    A survey of methods and codes which describe the flow of groundwater and the migration of radioactive waste in and about nuclear repositories was performed. A number of laboratories engaged in studies of waste migration and groundwater flow were visited in order to discuss the general problem and obtain reports of work being performed. The results of this survey are discussed

  15. Experimental study of water flow in nuclear fuel elements; Estudo experimental do escoamento de agua em elementos combustiveis nucleares

    Energy Technology Data Exchange (ETDEWEB)

    Rodrigues, Lorena Escriche, E-mail: ler@cdtn.br [Centro Federal de Educacao Tecnologica de Minas Gerais (CEFET), Belo Horizonte, MG (Brazil); Rezende, Hugo Cesar; Mattos, Joao Roberto Loureiro de; Barros Filho, Jose Afonso; Santos, Andre Augusto Campagnole dos, E-mail: hcr@cdtn.br, E-mail: jrmattos@cdtn.br, E-mail: jabf@cdtn.br, E-mail: aacs@cdtn.br [Centro de Desenvolvimento da Tecnologia Nuclear (CDTN/CNEN-MG), Belo Horizonte, MG (Brazil)

    2013-07-01

    This work aims to develop an experimental methodology for investigating the water flow through rod bundles after spacer grids of nuclear fuel elements of PWR type reactors. Speed profiles, with the device LDV (Laser Doppler Velocimetry), and the pressure drop between two sockets located before and after the spacer grid, using pressure transducers were measured.

  16. On the elliptic flow for nearly symmetric collisions and nuclear ...

    Indian Academy of Sciences (India)

    In figure 1, the final-state elliptic flow is displayed for the free particles (upper panel), light mass fragments (LMFs) (2 ≤ A ≤ 4) (middle panel) and intermediate mass fragments. Figure 1. The elliptic flow as a function of the transverse momentum at incident energy. 95 MeV/nucleon for free particles, LMFs and IMFs in top, ...

  17. CFD modelling of subcooled flow boiling for nuclear engineering applications

    International Nuclear Information System (INIS)

    Koncar, B.; Krepper, E.; Egorov, Y.

    2005-01-01

    In this work a general-purpose CFD code CFX-5 was used for simulations of subcooled flow boiling. The subcooled boiling model, available in a custom version of CFX-5, uses a special treatment of the wall boiling boundary, which assures the grid invariant solution. The simulation results have been validated against the published experimental data [1] of high-pressure flow boiling in a vertical pipe covering a wide range of conditions (relevant to the pressurized water reactor). In general, a good agreement with the experimental data has been achieved. To adequately predict the lateral distribution of two-phase flow parameters, the modelling of two-phase flow turbulence and non-drag forces under wall boiling conditions have been also investigated in the paper. (author)

  18. At the heart of the global nuclear news flow

    International Nuclear Information System (INIS)

    Feuz, P.

    1997-01-01

    The dual role of NUCNET is discussed: - exchange of news and other information within the world's nuclear community for use by top managers and executives and public communicators, and - distribution of news of interest to the public to wire agency journalists and other sections of the media

  19. Gas flow in and out of a nuclear waste container

    International Nuclear Information System (INIS)

    Zwahlen, E.D.; Pigford, T.H.; Chambre, P.L.; Lee, W.W.L.

    1989-05-01

    We analyze the flow of gases out of and into a high-level-waste container in the unsaturated tuff of Yucca Mountain. Containers are expected to fail eventually by localized cracks and penetrations. Even though the penetrations may be small, argon gas initially in the hot container can leak out. As the waste package cools, the pressure inside the container can become less than atmospheric, and air can leak in. 14 C released from the hot fuel-cladding surface can leak out of penetrations, and air inleakage can mobilize additional 14 C and other volatile radioactive species as it oxidizes the fuel cladding and the spent fuel. In an earlier paper we studied the gas flow through container penetrations occurring at the time of emplacement. Here we analyze the flow of gas for various penetration sizes occurring at 300 years. 3 refs., 2 figs

  20. Low-flow operation and testing of pumps in nuclear plants

    International Nuclear Information System (INIS)

    Greenstreet, W.L.

    1989-01-01

    Low-flow operation of centrifugal pumps introduces hydraulic instability and other factors that can cause damage to these machines. The resulting degradation has been studied and recorded for pumps in electric power plants. The objectives of this paper are to (1) describe the damage-producing phenomena, including their sources and consequences; (2) relate these observations to expectations for damage caused by low-flow operation of pumps in nuclear power plants; and (3) assess the utility of low-flow testing. Hydraulic behavior during low-flow operation is reviewed for a typical centrifugal pump stage, and the damage-producing mechanisms are described. Pump monitoring practices, in conjunction with pump performance characteristics, are considered; experience data are reviewed; and the effectiveness of low-flow surveillance monitoring is examined. Degradation caused by low-flow operation is shown to be an important factor, and low-flow surveillance testing is shown to be inadequate. 18 refs., 5 figs., 4 tabs

  1. 3-D flow analyses for design of nuclear fuel spacer

    Energy Technology Data Exchange (ETDEWEB)

    Karouta, Z. [ABB Combustion Engineering, Windsor, CT (United States); GU, Chun-Yuan [ABB Corporate Research, Vaesteras (Sweden); Schoelin, B. [ABB Atom AB, Vaesteras (Sweden)

    1995-09-01

    The Computational Fluid Dynamics (CFD) code, CFDS-FLOW3D, was used to develop improved fuel designs for PWR cores. It was used primarily to understand the fluid dynamics of grid spacers, the mass transfer between subchannels caused by spacers and in the long term to develop two-phase models which enable prediction of critical heat flux in PWR fuel. A single subchannel of one grid span was modeled. In this model different spacer designs with mixing devices were analyzed. A special treatment of the boundary condition was developed making use of flow symmetry to model the mass transfer between different subchannels and minimize the size of the computational model. This reduced the computational model to a fraction of a subchannel using traditional periodic boundary conditions. The Navier-Stokes equation was solved for the liquid and the flow turbulence was modeled by k-{xi} turbulence model. The spacer and mixing device were treated as infinite thin surfaces in the model and a zero velocity condition and turbulent wall function were applied on each side of the thin surfaces. This approach simulated the swirl from the mixing devices well, but had the drawback of not predicting pressure drop accurately since the wake behind the plates and the acceleration effect of the spacers were ignored. CFDS-FLOW3D models with mixing devices were applied in the single-phase flow regime. Velocity profiles from the CFDS-FLOW3D models were compared to Laser Doppler Velocimeter measurements taken from the flow field downstream of spaces in a full scale, cold water test loop. The predicted axial and lateral velocity profiles were in good agreement with the measurements. The evaluation of the performance of different spacer devices was made by comparing the swirl ratio downstream of the grid spacers. It is planned to evaluate heat transfer coefficient downstream of the spaces, to implement two-phase flow models, and to model the superheated boundary layer on the surface of the fuel rod.

  2. Nuclear reactor core supports, and flow control system

    International Nuclear Information System (INIS)

    1976-01-01

    This reactor core supporting structure consists of a single plate provided with canals situated under each fuel element for the crossing of the coolant. Each canal has a constricted intake part and a zone of larger diameter downstream this inlet port, in order for the coolant flow to be laminar at the canal outlets, and to be uniformly distributed inside the fuel elements [fr

  3. A tool for visualization of two-phase flow simulations related to nuclear safety

    International Nuclear Information System (INIS)

    Hyvaerinen, J.

    1991-01-01

    This paper describes a new tool, BOXER, that has been developed to produce animated visualization of data from computer simulations of two-phase (multi-component) flow phenomena in nuclear reactor systems. In the first part of the paper, background information regarding the type and the scope of the simulations is presented. The second part describes the tool, giving an example of its usage. BOXER has been developed at the Finnish Centre for Radiation and Nuclear Safety. (author)

  4. Triton-3He relative and differential flows and the high density behavior of nuclear symmetry

    International Nuclear Information System (INIS)

    Yong, Gaochan; Li, Baoan; Chen, Liewen

    2010-01-01

    Using a transport model coupled with a phase-space coalescence after-burner we study the triton- 3 He relative and differential transverse flows in semi-central 132 Sn + 124 Sn reactions at a beam energy of 400 MeV/nucleon. We find that the triton- 3 He pairs carry interesting information about the density dependence of the nuclear symmetry energy. The t- 3 He relative flow can be used as a particularly powerful probe of the high-density behavior of the nuclear symmetry energy. (author)

  5. Multiparameter cytometric analysis of complex cellular response

    Czech Academy of Sciences Publication Activity Database

    Šimečková, Šárka; Fedr, Radek; Remšík, Jan; Kahounová, Z.; Slabáková, Eva; Souček, Karel

    2018-01-01

    Roč. 93A, č. 2 (2018), s. 239-248 ISSN 1552-4922 R&D Projects: GA MZd(CZ) NV15-28628A; GA MZd(CZ) NV15-33999A; GA MZd(CZ) NV17-28518A Institutional support: RVO:68081707 Keywords : flow-cytometry * permeabilization * apoptosis * fixation Subject RIV: EB - Genetics ; Molecular Biology OBOR OECD: Cell biology Impact factor: 3.222, year: 2016

  6. Personal view of educating two-phase flow and human resource development as a nuclear engineer

    International Nuclear Information System (INIS)

    Hotta, Akitoshi

    2010-01-01

    As an engineer who has devoted himself in the nuclear industry for almost three decades, the author gave a personal view on educating two-phase flow and developing human resources. An expected role of universities in on-going discussions of collaboration among industry-government-academia is introduced. Reformation of two-phase flow education is discussed from two extreme viewpoints, the basic structure of physics and the practical system analysis. (author)

  7. Thermodynamic coupling of heat and matter flows in near-field regions of nuclear waste repositories

    International Nuclear Information System (INIS)

    Carnahan, C.L.

    1983-11-01

    In near-field regions of nuclear waste repositories, thermodynamically coupled flows of heat and matter can occur in addition to the independent flows in the presence of gradients of temperature, hydraulic potential, and composition. The following coupled effects can occur: thermal osmosis, thermal diffusion, chemical osmosis, thermal filtration, diffusion thermal effect, ultrafiltration, and coupled diffusion. Flows of heat and matter associated with these effects can modify the flows predictable from the direct effects, which are expressed by Fourier's law, Darcy's law, and Fick's law. The coupled effects can be treated quantitatively together with the direct effects by the methods of the thermodynamics of irreversible processes. The extent of departure of fully coupled flows from predictions based only on consideration of direct effects depends on the strengths of the gradients driving flows, and may be significant at early times in backfills and in near-field geologic environments of repositories. Approximate calculations using data from the literature and reasonable assumptions of repository conditions indicate that thermal-osmotic and chemical-osmotic flows of water in semipermeable backfills may exceed Darcian flows by two to three orders of magnitude, while flows of solutes may be reduced greatly by ultrafiltration and chemical osmosis, relative to the flows predicted by advection and diffusion alone. In permeable materials, thermal diffusion may contribute to solute flows to a smaller, but still significant, extent

  8. Experiments on one-phase thermally stratified flows in nuclear reactor pipe lines

    International Nuclear Information System (INIS)

    Rezende, Hugo Cesar; Navarro, Moyses Alberto; Jordao, Elizabete; Santos, Andre Augusto Campagnole dos

    2009-01-01

    The phenomenon of thermal stratified flows occurs when two different layers of the same liquid at different temperatures flow separately in horizontal pipes without appreciable mixing. This phenomenon was not considered in the design stage of most of the operating nuclear power plants, but in last two decades it has become apparent due to the temperature monitoring of piping systems. The occurrence of temperature differences of about 200 deg C have been found in a narrow band around the hot and cold water interface in components under stratified flows. Loadings due to thermal stratification affected the integrity of safety related piping systems. This paper presents the results of a range of experiments performed to simulate one phase thermally stratified flows in geometry and flow condition representing a nuclear reactor steam generator injection nozzle. They have the objective of studying the flow configurations and understanding the evolution of the thermal stratification process. The driving parameter considered to characterize flow under stratified regime due to difference in specific masses is the Froude number. Different Froude numbers, from 0.018 to 0.22, were obtained in different testes by setting injection cold water flow rates and hot water initial temperatures as planned in the test matrix. Results are presented showing the influence of Froude number on the hot and cold water interface position, temperature gradients and striping phenomenon. (author)

  9. Application of neural networks to validation of feedwater flow rate in a nuclear power plant

    International Nuclear Information System (INIS)

    Khadem, M.; Ipakchi, A.; Alexandro, F.J.; Colley, R.W.

    1993-01-01

    Feedwater flow rate measurement in nuclear power plants requires periodic calibration. This is due to the fact that the venturi surface condition of the feedwater flow rate sensor changes because of a chemical reaction between the surface coating material and the feedwater. Fouling of the venturi surface, due to this chemical reaction and the deposits of foreign materials, has been observed shortly after a clean venturi is put in operation. A fouled venturi causes an incorrect measurement of feedwater flow rate, which in turn results in an inaccurate calculation of the generated power. This paper presents two methods for verifying incipient and continuing fouling of the venturi of the feedwater flow rate sensors. Both methods are based on the use of a set of dissimilar process variables dynamically related to the feedwater flow rate variable. The first method uses a neural network to generate estimates of the feedwater flow rate readings. Agreement, within a given tolerance, of the feedwater flow rate instrument reading, and the corresponding neural network output establishes that the feedwater flow rate instrument is operating properly. The second method is similar to the first method except that the neural network predicts the core power which is calculated from measurements on the primary loop, rather than the feedwater flow rates. This core power is referred to the primary core power in this paper. A comparison of the power calculated from the feedwater flow measurements in the secondary loop, with the calculated and neural network predicted primary core power provides information from which it can be determined whether fouling is beginning to occur. The two methods were tested using data from the feedwater flow meters in the two feedwater flow loops of the TMI-1 nuclear power plant

  10. Comparison of four nuclear isolation buffers for plant DNA flow cytometry

    Czech Academy of Sciences Publication Activity Database

    Loureiro, J.; Rodriguez, E.; Doležel, Jaroslav; Santos, C.

    2006-01-01

    Roč. 98, - (2006), s. 679-689 ISSN 0305-7364 R&D Projects: GA MŠk(CZ) LC06004 Institutional research plan: CEZ:AV0Z50380511 Keywords : flow cytometry * genome size * nuclear DNA content Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.448, year: 2006

  11. Flow: Statistics, visualization and informatics for flow cytometry

    Directory of Open Access Journals (Sweden)

    Kepler Thomas B

    2008-06-01

    Full Text Available Abstract Flow is an open source software application for clinical and experimental researchers to perform exploratory data analysis, clustering and annotation of flow cytometric data. Flow is an extensible system that offers the ease of use commonly found in commercial flow cytometry software packages and the statistical power of academic packages like the R BioConductor project.

  12. Monitoring functions in managed microbial systems by cytometric bar coding.

    Science.gov (United States)

    Koch, Christin; Fetzer, Ingo; Schmidt, Thomas; Harms, Hauke; Müller, Susann

    2013-02-05

    Cytometric monitoring of microbial community dynamics can be used to estimate stability of technical microbial processes like biogas production by analysis of segregated cell abundance changes. In this study, structure variations of a biogas community were cytometrically recorded over 9 months and found to be of diagnostic value for process details. The reactor regime was intentionally disturbed with regard to substrate overload or H(2)S accumulation. A single-cell based approach called cytometric bar coding (CyBar) for fast identification of reactive subcommunities was used. Functionality of specific subcommunities was uncovered by processing CyBar data with abiotic reactor parameters using Spearman's correlation coefficient. Twenty subcommunities showed a discrete and divergent behavior. For example, a 4-fold substrate overload increased the cell number of two acidogenic index subcommunities to 176 and 193% within three days. Supplementary analyses were done using DNA fingerprinting, cloning, and sequencing. Bioreactor perturbations were shown to create cell abundance changes in subcommunities rather than variations in their phylogenetic composition. The used workflow and macros are ready-to-use tools and allow on-site monitoring and interpretation of variation in microbial community functions within a few hours.

  13. Phenomenological studies of two-phase flow processes for nuclear waste isolation

    International Nuclear Information System (INIS)

    Pruess, K.; Finsterle, S.; Persoff, P.; Oldenburg, C.

    1994-01-01

    The US civilian radioactive waste management program is unique in its focus on a site in the unsaturated zone, at Yucca Mountain, Nevada. Two-phase flow phenomena can also play an important role in repositories beneath the water table where gas is generated by corrosion, hydrolysis, and biological degradation of the waste packages. An integrated program has been initiated to enhance our understanding of two-phase flow behavior in fractured rock masses. The studies include two-phase (gas-liquid) flow experiments in laboratory specimens of natural rock fractures, analysis and modeling of heterogeneity and instability effects in two-phase flow, and design and interpretation of field experiments by means of numerical simulation. We present results that identify important aspects of two-phase flow behavior on different space and time scales which are relevant to nuclear waste disposal in both unsaturated and saturated formations

  14. Investigation of fluid flow in various geometries related to nuclear reactor using PIV system

    International Nuclear Information System (INIS)

    Kansal, A.K.; Maheshwari, N.K.; Singh, R.K.; Vijayan, P.K.; Saha, D.; Singh, R.K.; Joshi, V.M.

    2011-01-01

    Particle Image Velocimetry (PIV) is a non-intrusive technique for simultaneously measuring the velocities at many points in a fluid flow. The PIV system used is comprised of Nd:YAG laser source, CCD (Charged Coupled Device) camera, timing controller (to control the laser and camera) and software used for analyzing the flow velocities. Several case studies related to nuclear reactor were performed with the PIV system. Some of the cases like flow in circular tube, submerged jet, natural convection in a water pool, flow field of moderator inlet diffuser of 500 MWe Pressurised Heavy Water Reactor (PHWR) and fluidic flow control device (FFCD) used in advanced accumulator of Emergency Core Cooling System (ECCS) have been studied using PIV system. Theoretical studies have been performed and comparisons with PIV results are also given in the present studies. (author)

  15. Multiparameter cytometric analysis of complex cellular response.

    Science.gov (United States)

    Šimečková, Šárka; Fedr, Radek; Remšík, Ján; Kahounová, Zuzana; Slabáková, Eva; Souček, Karel

    2018-02-01

    Complex analysis of cellular responses after experimental treatment is important for screening, mechanistic understanding of treatment effects, and the identification of sensitive and resistant cell phenotypes. Modern multicolor flow cytometry has demonstrated its power for such analyses. Here, we introduce a multiparametric protocol for complex analysis of cytokinetics by the simultaneous detection of seven fluorescence parameters. This analysis includes the detection of two surface markers for immunophenotyping, analysis of proliferation based on the cell cycle and the measurement of incorporated nucleoside analogue 5-ethynyl-2'-deoxyuridine (EdU) in newly synthesized DNA, analysis of DNA damage using an anti-phospho-histone H2A.X (Ser139) antibody, and determination of cell death using a fixable viability probe and intracellular detection of caspase-3 activation. To demonstrate the applicability of this protocol for the analysis of heterogeneous and complex cell responses, we used different treatments and model cell lines. We demonstrated that this protocol has the potential to provide complex and simultaneous analysis of cytokinetics and analyze the heterogeneity of the response at the single-cell level. © 2017 International Society for Advancement of Cytometry. © 2017 International Society for Advancement of Cytometry.

  16. Coupled analysis of flow and heat around a high-level nuclear waste repository

    International Nuclear Information System (INIS)

    Utsugida, Y.

    1985-01-01

    This paper presents the formulation of coupled flow and heat models by F.E.M. and the results of two kinds of analysis around a high-level nuclear waste repository. A one-dimensional element is used in modeling fluid flow in fractured media, but two-dimensional elements are used in modeling fluid flow and heat transport in porous media. A 2-order Runge-Kutta scheme is employed to carry out the time integration, and the coefficients of the capacity matrix are lumped at the nodes. One of the analyses is in the near field which regards disposal tunnel scale. The effects of the fluid flow are investigated on the temperature distribution in the vicinity of a disposal tunnel. The other is in the far field which regards the regional scale of the entire repository. The effects of radio-genic heat being produced by the nuclear wastes are investigated on the ground water flow in the cases of major fractured rock and no major fractured rock. In conclusion, there is only a slight influence from fluid flow in the near field and from heat in the far field in the case of no major fractured rock. On the other hand, the influence of radio-genic heat in the far field in the case of major fractured rock can not be ignored

  17. Consideration of Typical Nuclear Power Plant Site Characteristics for Groundwater Flow

    International Nuclear Information System (INIS)

    Kim, Chan Hoi; Yee, Eric

    2014-01-01

    This is complicated by the extreme measures the plant owners have taken by pumping water into the disabled plant in order to cool the corium, making it difficult for workers to inspect the facility and mitigate radiation hazards. One of most confusing aspects of the groundwater contamination issue at the Fukushima Daiichi nuclear power plant is the flow of groundwater, with conflicting reports saying there is very little water to significant amounts of water flowing towards the plant. Initially, different media outlets, reports, and figures show anywhere from uniform flow to nearly impossible flow situations, with a general improvement in groundwater flow feasibility over time as perhaps more knowledge of the subgrade or facilities is revealed. This situation highlights the importance of groundwater models, which traditionally use averaged and macro-scale adjusted subgrade properties. These are not necessarily bad practices, but some details may be lost in the process for more local scenarios. Therefore, this paper focuses on the site conditions of a typical nuclear power plant and its influences on local groundwater flow modelling. Site-specific data from the hydrologic system investigation must be prepared and utilized to evaluate the existing groundwater conditions and to identify pathway of groundwater flow toward subsurface and plant facilities before and after nuclear power plant construction by installing monitoring wells. These investigation data, evaluation and identification provide the basis for developing an overall conceptual model of groundwater. With this conceptual model, assumption of radioactive material release, for instance, the liquid radioactive waste from a ruptured tank in the compound building through cracks in the foundation wall enter the groundwater system, can be evaluated

  18. Improved method for fluorescence cytometric immunohematology testing.

    Science.gov (United States)

    Roback, John D; Barclay, Sheilagh; Hillyer, Christopher D

    2004-02-01

    A method for accurate immunohematology testing by fluorescence cytometry (FC) was previously described. Nevertheless, the use of vacuum filtration to wash RBCs and a standard-flow cytometer for data acquisition hindered efforts to incorporate this method into an automated platform. A modified procedure was developed that used low-speed centrifugation of 96-well filter plates for RBC staining. Small-footprint benchtop capillary cytometers (PCA and PCA-96, Guava Technologies, Inc.) were used for data acquisition. Authentic clinical samples from hospitalized patients were tested for ABO group and the presence of D antigen (n = 749) as well as for the presence of RBC alloantibodies (n = 428). Challenging samples with mixed-field reactions and weak antibodies were included. Results were compared to those obtained by column agglutination technology (CAT), and discrepancies were resolved by standard tube methods. Detailed investigations of FC sensitivity and reproducibility were also performed. The modified FC method with the PCA determined the correct ABO group and D type for 98.7 percent of 520 samples, compared to 98.8 percent for CAT (p > 0.05). No-type-determined (NTD) rates were 1.2 percent for both methods. In testing for unexpected alloantibodies, FC determined the correct result for 98.6 percent of 215 samples, compared to 96.3 percent for CAT (p > 0.05). When samples were automatically acquired in the 96-well plate format with the PCA-96, 98.7 percent of 229 samples had correct ABO group and D type determined by FC, compared to 97.4 percent for CAT (p > 0.05). NTD rates were 0.9 and 2.6 percent, respectively. Antibody screens were accurate for 99.1 percent of 213 samples with the PCA-96, compared to 99.5 percent for CAT (p > 0.05). Further investigations demonstrated that FC with the PCA-96 was better than CAT at detecting weak anti-A (p < 0.0001) and alloantibodies. An improved method for FC immunohematology testing has been described. This assay was comparable

  19. Analysis of nuclear localization of interleukin-1 family cytokines by flow cytometry.

    Science.gov (United States)

    Ross, Ralf; Grimmel, Jan; Goedicke, Sybelle; Möbus, Anna M; Bulau, Ana-Maria; Bufler, Philip; Ali, Shafaqat; Martin, Michael U

    2013-01-31

    The dual function cytokines IL-1α, IL-33 and IL-37 are members of the IL-1 cytokine family. Besides of being able to bind to their cognate receptors on target cells, they can act intracellularly in the producing cell. All three are able to translocate to the nucleus and have been discussed to affect gene expression. In order to compare and quantitate nuclear translocation of these IL-1 family members we established a robust technique which enables to measure nuclear localization on a single cell level by flow cytometry. Vectors encoding fusion proteins of different IL-1 family members with enhanced green fluorescent protein were cloned and cell lines transiently transfected with these. Fluorescent fusion proteins in intact cells or in isolated nuclei were detected subsequently by fluorescence microscopy and flow cytometry, respectively. Depending on the cellular system, cells and nuclei were distinguishable by flow cytometry in forward scatter/sideward scatter. Fluorescent fusion proteins were detectable in isolated nuclei up to three days following preparation. Signal intensity of fusion proteins of IL-33 and IL-37 in isolated nuclei but not of IL-1α, was markedly increased by fixation with paraformaldehyde, directly following cell lysis, indicating that IL-1α binds stronger to nuclear structures than IL-33 and IL-37. Nuclear translocation of fluorescent IL-37 fusion proteins in a stably transfected RAW264.7 mouse macrophage cell line required stimulation with lipopolysaccharide. Applying this method we demonstrated that a prolonged lag phase of more than 15h before LPS-stimulated nuclear translocation was detected. In summary, we present a robust method to analyze and quantitate nuclear localization of IL-1 cytokine family members. Copyright © 2012 Elsevier B.V. All rights reserved.

  20. Nuclear DNA content of the pigeon orchid (Dendrobium crumenatum Sw.) with the analysis of flow cytometry

    OpenAIRE

    Upatham Meesawat; Theera Srisawat; Ladda Eksomtramage; Kamnoon Kanchanapoom

    2008-01-01

    Nuclear DNA content for the adult plants grown in a greenhouse and in vitro young plantlets of the pigeon orchid (Dendrobium crumenatum Sw.) was analyzed using flow cytometry. The resulting 2C DNA values ranged from 2.30±0.14 pgto 2.43±0.06 pg. However, nuclear DNA ploidy levels of long-term in vitro plantlets were found to be triploid and tetraploid.These ploidy levels were confirmed by chromosome counting. Tetraploid individuals (2n = 4x = 76) had approximately two times DNA content than di...

  1. Steady MHD flow for molten metal coolants in advanced nuclear systems

    International Nuclear Information System (INIS)

    Koreshi, Z.U.

    2004-01-01

    The flow of a molten metal coolant, incorporating viscosity and a magnetic force in a channel typical of a nuclear fusion reactor is formulated. This is based on the flow conservation equations (continuity and momentum), along with Maxwell's equations and Ohm's law. These equations are solved with assumptions of steady fully-developed flow for Newtonian fluids. Exact solutions are compared with a Direct Numerical Simulation (DNS) to compute important engineering design parameters such as velocity distribution and pressure drop in a coolant channel. The analysis can be used for several candidate coolants including, molten lithium, sodium, lead, and compounds. The difference in velocity profile, and hence pressure drop, is strongly dependent on the magnetic field. In fact, it is shown that a flat (almost-turbulent) profile is obtained for the flow. (author)

  2. An application of multilevel flow modelling method for nuclear plant state identification

    International Nuclear Information System (INIS)

    Businaro, T.; Di Lorenzo, A.; Meo, G.B.; Rabbani, M.I.; Rubino, E.

    1986-01-01

    With the advent of advanced digital techniques it has been rendered possible, necessity of which has long since been recognised, to develop a computer based man-machine interface and hance an expert system based on knowledge based decision making for operator support in the control rooms of nuclear plants. The Multilevel Flow Modelling method developed at RISO Laboratories, Denmark, has been applied in the present experiment to model Italian PEC reactor and to verify applicability of this method in plant state identification. In MFM method functional structure of a process plant is described in terms of a set of interrelated mass and energy flow structures on different levels of physical aggregation

  3. Control flow of radiopharmaceuticals in nuclear medicine by means of an E-service

    International Nuclear Information System (INIS)

    Nunez Martin, L.; Gonzalez de Mingo, M. A.; Fragua Redondo, J. A.; Martinez Ortega, J.; Gutierrez Camunas, S.; Redondo Miguel, A. B.

    2013-01-01

    The almost generalized use of single-dose Nuclear Medicine for performing diagnostic tests or treatments, and the consequent complexity that accompanies its management, has resulted in the need to control the flow of material radioisotopic tools. An e-service is designed to manage the flow of radiopharmaceuticals and control its use and spending. This control does not only affect the efficiency in the use and cost of material, but in the radioactive waste associated with the non-use and waste reduction and a more effective organization of the Department. (Author)

  4. Visual Inspection of the Flow Distribution Plate Bolts of a Nuclear Steam Generator

    International Nuclear Information System (INIS)

    Jeong, Woo Tae; Kim, Suk Tae; Sohn, Wook; Kang, Duk Won; Kang, Seok Chul

    2007-01-01

    To develop a system for visually inspecting the flow distribution plate (FDP) bolts of a nuclear steam generator, we reviewed several types of similar inspection equipment. The equipment which are currently available are mostly for inspecting lower part of a steam generator such as tube sheets and annulus except ELVS (Eggcrate Visual Inspection System). However, the design concept of ELVS could not be used for developing a device which enables the visual inspection of flow distribution plate bolts. Therefore, based on the current state of the art technology on the similar equipment, we conceptually designed a new inspection system for checking the FDP bolts

  5. Nuclear-radiation-actuated valve. [Patent application; for increasing coolant flow to blanket

    Science.gov (United States)

    Christiansen, D.W.; Schively, D.P.

    1982-01-19

    The present invention relates to a breeder reactor blanket fuel assembly coolant system valve which increases coolant flow to the blanket fuel assembly to minimize long-term temperature increases caused by fission of fissile fuel created from fertile fuel through operation of the breeder reactor. The valve has a valve first part (such as a valve rod with piston) and a valve second part (such as a valve tube surrounding the valve rod, with the valve tube having side slots surrounding the piston). Both valve parts have known nuclear radiation swelling characteristics. The valve's first part is positioned to receive nuclear radiation from the nuclear reactor's fuel region. The valve's second part is positioned so that its nuclear radiation induced swelling is different from that of the valve's first part. The valve's second part also is positioned so that the valve's first and second parts create a valve orifice which changes in size due to the different nuclear radiation caused swelling of the valve's first part compared to the valve's second part. The valve may be used in a nuclear reactor's core coolant system.

  6. New developments in two-phase flow heat transfer with emphasis on nuclear safety research

    International Nuclear Information System (INIS)

    Mayinger, F.

    1987-01-01

    The literature on two-phase flow - with and without heat transfer - shows an explosive-like growth of published papers within the last ten years. Many of these papers were published as a result of nuclear safety research. It is impossible to deal with all new developments reported in this extensive literature. So one has to ask: Are there trends of special interest, where this report could be concentrated on? Looking over the situation, there seem to be three very promising fields of research having high actuality, especially for nuclear safety, namely: fluiddynamic and thermodynamic nonequilibrium in steady state, transient conditions, and scaling. The discussion on new developments in two-phase flow heat transfer, therefore, is limited on these subjects

  7. Possibilities and Limitations of CFD Simulation for Flashing Flow Scenarios in Nuclear Applications

    OpenAIRE

    Liao, Yixiang; Lucas, Dirk

    2017-01-01

    The flashing phenomenon is relevant to nuclear safety analysis, for example by a loss of coolant accident and safety release scenarios. It has been studied intensively by means of experiments and simulations with system codes, but computational fluid dynamics (CFD) simulation is still at the embryonic stage. Rapid increasing computer speed makes it possible to apply the CFD technology in such complex flow situations. Nevertheless, a thorough evaluation on the limitations and restrictions is s...

  8. Evaluation of groundwater flow and transport at the Shoal underground nuclear test: An interim report

    Energy Technology Data Exchange (ETDEWEB)

    Pohll, G.; Chapman, J.; Hassan, A.; Papelis, C.; Andricevic, R.; Shirley, C.

    1998-07-01

    Since 1962, all United States nuclear tests have been conducted underground. A consequence of this testing has been the deposition of large amounts of radioactive materials in the subsurface, sometimes in direct contact with groundwater. The majority of this testing occurred on the Nevada Test Site, but a limited number of experiments were conducted in other locations. One of these is the subject of this report, the Project Shoal Area (PSA), located about 50 km southeast of Fallon, Nevada. The Shoal test consisted of a 12-kiloton-yield nuclear detonation which occurred on October 26, 1963. Project Shoal was part of studies to enhance seismic detection of underground nuclear tests, in particular, in active earthquake areas. Characterization of groundwater contamination at the Project Shoal Area is being conducted by the US Department of Energy (DOE) under the Federal Facility Agreement and Consent Order (FFACO) with the State of Nevada Department of Environmental Protection and the US Department of Defense (DOD). This order prescribes a Corrective Action Strategy (Appendix VI), which, as applied to underground nuclear tests, involves preparing a Corrective Action Investigation Plan (CAIP), Corrective Action Decision Document (CADD), Corrective Action Plan, and Closure Report. The scope of the CAIP is flow and transport modeling to establish contaminant boundaries that are protective of human health and the environment. This interim report describes the current status of the flow and transport modeling for the PSA.

  9. Vertical and longitudinal gradients in HNA-LNA cell abundances and cytometric characteristics in the Mediterranean Sea

    Directory of Open Access Journals (Sweden)

    F. Van Wambeke

    2011-07-01

    Full Text Available Heterotrophic bacterioplankton abundance and production were investigated with depth (down to bathypelagic layers and with longitude (from 4.9° E to 32.7° E along a cruise track across the Mediterranean Sea in early summer 2008. Abundances and flow cytometric characteristics (green fluorescence and side scatter signals of high nucleic acid (HNA and low nucleic acid (LNA bacterial cells were determined using flow cytometry. Contrary to what is generally observed, the relative importance of HNA cells, as a percent of total cells, (%HNA, range 30–69 % was inversely related to bacterial production (range 0.15–44 ng C l−1 h−1 although the negative relation was weak (log–log regression r2=0.19. The %HNA as well as the mean side scatter of HNA group increased significantly with depth in the meso and bathypelagic layers. Vertical stratification played an important role in influencing the distribution and characteristics of bacterial cells especially with regard to layers located above, within or below the deep chlorophyll maximum. Within a given layer, the relationships between the flow cytometric characteristics and environmental variables such as chlorophyll-a, nutrients or bacterial production changed. Overall, the relationships between HNA and LNA cells and environmental parameters differed vertically more than longitudinally.

  10. Accident-induced flow and material transport in nuclear facilities: a literature review

    International Nuclear Information System (INIS)

    Bolstad, J.W.; Gregory, W.S.; Martin, R.A.; Tang, P.K.; Merryman, R.G.; Novat, J.; Whitmore, H.L.

    1984-04-01

    The reported investigation is part of a program that was established for deriving radiological source terms at a nuclear facility's atmospheric boundaries under postulated accident conditions. The overall program consists of three parts: (1) accident delineation and survey, (2) internal source term characterization and release, and (3) induced flow and material transport. This report is an outline of pertinent induced-flow and material transport literature. Our objectives are to develop analytical techniques and data that will permit prediction of accident-induced transport of airborne material to a plant's atmospheric boundaries. Prediction of material transport requires investigation of the areas of flow dynamics and reentrainment/deposition. A review of material transport, fluid dynamics, and reentrainment/deposition literature is discussed. In particular, those references dealing with model development are discussed with special emphasis on application to a facility's interconnected ventilation system. 176 references

  11. Modeling studies of multiphase fluid and heat flow processes in nuclear waste isolation

    International Nuclear Information System (INIS)

    Pruess, K.

    1989-01-01

    Multiphase fluid and heat flow plays an important role in many problems relating to the disposal of nuclear wastes in geologic media. Examples include boiling and condensation processes near heat-generating wastes, flow of water and formation gas in partially saturated formations, evolution of a free gas phase from waste package corrosion in initially water-saturated environments, and redistribution (dissolution, transport and precipitation) of rock minerals in non-isothermal flow fields. Such processes may strongly impact upon waste package and repository design considerations and performance. This paper summarizes important physical phenomena occurring in multiphase and nonisothermal flows, as well as techniques for their mathematical modeling and numerical simulation. Illustrative applications are given for a number of specific fluid and heat flow problems, including: thermohydrologic conditions near heat-generating waste packages in the unsaturated zone; repositorywide convection effects in the unsaturated zone; effects of quartz dissolution and precipitation for disposal in the saturated zone; and gas pressurization and flow effects from corrosion of low-level waste packages

  12. Modeling studies for multiphase fluid and heat flow processes in nuclear waste isolation

    International Nuclear Information System (INIS)

    Pruess, K.

    1988-07-01

    Multiphase fluid and heat flow plays an important role in many problems relating to the disposal of nuclear wastes in geologic media. Examples include boiling and condensation processes near heat-generating wastes, flow of water and formation gas in partially saturated formations, evolution of a free gas phase from waste package corrosion in initially water-saturated environments, and redistribution (dissolution, transport, and precipitation) of rock minerals in non-isothermal flow fields. Such processes may strongly impact upon waste package and repository design considerations and performance. This paper summarizes important physical phenomena occurring in multiphase and nonisothermal flows, as well as techniques for their mathematical modeling and numerical simulation. Illustrative applications are given for a number of specific fluid and heat flow problems, including: thermohydrologic conditions near heat-generating waste packages in the unsaturated zone; repository-wide convection effects in the unsaturated zone; effects of quartz dissolution and precipitation for disposal in the saturated zone; and gas pressurization and flow corrosion of low-level waste packages. 34 refs; 7 figs; 2 tabs

  13. Transport of nuclear waste flows - a modelling and simulation approach - 59136

    International Nuclear Information System (INIS)

    Adams, Jonathan F.W.; Biggs, Simon R.; Fairweather, Michael; Yao, Jun; Young, James

    2012-01-01

    The task of implementing safer and more efficient processing and transport techniques in the handling of nuclear wastes made up of liquid-solid mixtures provides a challenging and interesting area of research. The radioactive nature of nuclear waste means that it is difficult to perform experimental studies of its transport. In contrast, the use of modelling and simulation techniques can help to elucidate the physics that underpin such flows and provide valuable insights into common problems associated with their transport, as well as assisting in the focusing of experimental research. Two phase solid-liquid waste-forms are commonplace within the nuclear reprocessing industry. Currently, there is waste, e.g., in the form of a solid-liquid slurry in cooling ponds and liquid flows containing suspensions of solid particles feature heavily in the treatment and disposal of this waste. With nuclear waste in the form of solid-liquid sludges it is important to understand the nature of the flow, with particular interest in the settling characteristics of the particulate waste material. Knowledge of the propensity of pipe flows to form solid beds is important in avoiding unwanted blockages in pipelines and pumping systems. In cases where the formation of a solid bed is unavoidable, it is similarly important to know how the modified cross-sectional area of the pipe, due to the presence of a bed, will affect particle behaviour through the creation of secondary flows effects that are also common to square duct flows. A greater understanding of particle deposition in square ducts and pipes of circular cross-section is also of significant and broad industrial relevance, with flows containing particulates prevalent throughout the nuclear, pharmaceutical, chemical, mining and agricultural industries. A greater understanding of particle behaviour in square ducts and circular pipes with variable bed height is the focus of this current work. The more computationally expensive but

  14. Gene flow among wild and domesticated almond species: insights from chloroplast and nuclear markers

    Science.gov (United States)

    Delplancke, Malou; Alvarez, Nadir; Espíndola, Anahí; Joly, Hélène; Benoit, Laure; Brouck, Elise; Arrigo, Nils

    2012-01-01

    Hybridization has played a central role in the evolutionary history of domesticated plants. Notably, several breeding programs relying on gene introgression from the wild compartment have been performed in fruit tree species within the genus Prunus but few studies investigated spontaneous gene flow among wild and domesticated Prunus species. Consequently, a comprehensive understanding of genetic relationships and levels of gene flow between domesticated and wild Prunus species is needed. Combining nuclear and chloroplastic microsatellites, we investigated the gene flow and hybridization among two key almond tree species, the cultivated Prunus dulcis and one of the most widespread wild relative Prunus orientalis in the Fertile Crescent. We detected high genetic diversity levels in both species along with substantial and symmetric gene flow between the domesticated P. dulcis and the wild P. orientalis. These results were discussed in light of the cultivated species diversity, by outlining the frequent spontaneous genetic contributions of wild species to the domesticated compartment. In addition, crop-to-wild gene flow suggests that ad hoc transgene containment strategies would be required if genetically modified cultivars were introduced in the northwestern Mediterranean. PMID:25568053

  15. Modelling turbulent fluid flows in nuclear and fossil-fired power plants

    International Nuclear Information System (INIS)

    Viollet, P.L.

    1995-06-01

    The turbulent flows encountered in nuclear reactor thermal hydraulic studies or fossil-fired plant thermo-aerodynamic analyses feature widely varying characteristics, frequently entailing heat transfers and two-phase flows so that modelling these phenomena tends more and more to involve coupling between several branches of engineering. Multi-scale geometries are often encountered, with complex wall shapes, such as a PWR vessel, a reactor coolant pump impeller or a circulating fluidized bed combustion chamber. When it comes to validating physical models of these flows, the analytical process highlights the main descriptive parameters of local flow conditions: tensor characterizing the turbulence anisotropy, characteristic time scales for turbulent flow particle dynamics. Cooperative procedures implemented between national or international working parties can accelerate validation by sharing and exchanging results obtained by the various organizations involved. With this principle accepted, we still have to validate the products themselves, i.e. the software used for the studies. In this context, the ESTET, ASTRID and N3S codes have been subjected to a battery of test cases covering their respective fields of application. These test cases are re-run for each new version, so that the sets of test cases systematically benefit from the gradually upgraded functionalities of the codes. (author). refs., 3 figs., 6 tabs

  16. Experimental and computational investigation of flow of pebbles in a pebble bed nuclear reactor

    Science.gov (United States)

    Khane, Vaibhav B.

    The Pebble Bed Reactor (PBR) is a 4th generation nuclear reactor which is conceptually similar to moving bed reactors used in the chemical and petrochemical industries. In a PBR core, nuclear fuel in the form of pebbles moves slowly under the influence of gravity. Due to the dynamic nature of the core, a thorough understanding about slow and dense granular flow of pebbles is required from both a reactor safety and performance evaluation point of view. In this dissertation, a new integrated experimental and computational study of granular flow in a PBR has been performed. Continuous pebble re-circulation experimental set-up, mimicking flow of pebbles in a PBR, is designed and developed. Experimental investigation of the flow of pebbles in a mimicked test reactor was carried out for the first time using non-invasive radioactive particle tracking (RPT) and residence time distribution (RTD) techniques to measure the pebble trajectory, velocity, overall/zonal residence times, flow patterns etc. The tracer trajectory length and overall/zonal residence time is found to increase with change in pebble's initial seeding position from the center towards the wall of the test reactor. Overall and zonal average velocities of pebbles are found to decrease from the center towards the wall. Discrete element method (DEM) based simulations of test reactor geometry were also carried out using commercial code EDEM(TM) and simulation results were validated using the obtained benchmark experimental data. In addition, EDEM(TM) based parametric sensitivity study of interaction properties was carried out which suggests that static friction characteristics play an important role from a packed/pebble beds structural characterization point of view. To make the RPT technique viable for practical applications and to enhance its accuracy, a novel and dynamic technique for RPT calibration was designed and developed. Preliminary feasibility results suggest that it can be implemented as a non

  17. Nuclear power plant life management: flow accelerated corrosion and chemical control. Application to Embalse Nuclear power plant

    International Nuclear Information System (INIS)

    Chocron, Mauricio; Saucedo, Ramona E.; Sainz, Ricardo A.; Ovando, Luis E.

    2006-01-01

    The chemistry of a water-steam cycle is one of the main aspects of the Plant Life Management of a Nuclear Power Plant and it is important for the preservation, efficiency and availability of the whole system. In that sense this aspect has to be prioritized in any study whose aim is the life extension of the plant. In particular, the flow-assisted -corrosion or FAC is a problem that worldwide has been considered important due to the piping wall thinning that in some occasions has led to severe accidents. The FAC phenomena is not easy to be interpreted and addressed although nowadays there are some accepted models to understand and predict sensitive areas of the cycle. The objectives of the present paper have been: a) The construction of an integrated code that involves all the aspects that have influence on FAC, i.e., materials, composition, geometry, temperature and flow rate, quality, chemistry, etc.; b) Establish or adapting current models to the circuit of Embalse PHWR NPP; c) Identify new locations for inspection and wall thickness measurement in order to predict residual life; d) Compare different chemistries and e) handle large sets of inspection data. Among the results, new lines have been incorporated to the inspection schedule of the 2005' programmed outage. Also, the evaluation is part of the PLIM-PLEX programme at Embalse-N.A.S.A. in collaboration with C.N.E.A. is being carried out. (author)

  18. Determination of velocity and flow direction of ground water by using nuclear techniques

    International Nuclear Information System (INIS)

    Santos Ferreira, L. dos.

    1976-06-01

    The dynamics of water in an aquifer with the purpose of determining the filtration velocity and the direction of groundwater flow with radioactive tracers was studied. Field equipment for the purposes of the study was built in the Laboratory of Tracers in Hydrology in collaboration with the Institute of Nuclear Engineering (IEN/NUCLEBRAS). The equipment was designed to minimize the possible vertical flows, loss and molecular diffusion of the tracer out of the studied region. The performance of the nuclear detectors and the constructional details of the field equipament were examined. The selection of the radioactive tracers was made taking into account its availibility and radiation facilities, cost of the inactive material and their physical and chemical properties. The tracers used were 82 Br and 198 Au. The results are discussed with the help of auxiliary informations such as plots of water levels in time and space, profiles and grain analysis. In order to obtain a physical explanation of the results, a qualitative model of the flow in the aquifer is also presented. (Author) [pt

  19. Development of an Efficient Meso- scale Multi-phase Flow Solver in Nuclear Applications

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Taehun [City Univ. (CUNY), NY (United States)

    2015-10-20

    The proposed research aims at formulating a predictive high-order Lattice Boltzmann Equation for multi-phase flows relevant to nuclear energy related application - namely, saturated and sub-cooled boiling in reactors, and liquid- liquid mixing and extraction for fuel cycle separation. An efficient flow solver will be developed based on the Finite Element based Lattice Boltzmann Method (FE- LBM), accounting for phase-change heat transfer and capable of treating multiple phases over length scales from the submicron to the meter. A thermal LBM will be developed in order to handle adjustable Prandtl number, arbitrary specific heat ratio, a wide range of temperature variations, better numerical stability during liquid-vapor phase change, and full thermo-hydrodynamic consistency. Two-phase FE-LBM will be extended to liquid–liquid–gas multi-phase flows for application to high-fidelity simulations building up from the meso-scale up to the equipment sub-component scale. While several relevant applications exist, the initial applications for demonstration of the efficient methods to be developed as part of this project include numerical investigations of Critical Heat Flux (CHF) phenomena in nuclear reactor fuel bundles, and liquid-liquid mixing and interfacial area generation for liquid-liquid separations. In addition, targeted experiments will be conducted for validation of this advanced multi-phase model.

  20. Nuclear DNA content of the hybrid plant pathogen Phytophthora andina determined by flow cytometry.

    Science.gov (United States)

    Wang, Jianan; Presser, Jackson W; Goss, Erica M

    2016-09-01

    Phytophthora andina is a heterothallic plant pathogen of Andean solanaceous hosts and is an interspecific hybrid of P. infestans and an unknown Phytophthora species. The objective of this study was to estimate the nuclear DNA content of isolates in three clonal lineages of P. andina relative to P. infestans Twelve isolates of P. andina and six isolates of P. infestans were measured for nuclear DNA content by propidium iodide-stained flow cytometry. We found that the DNA content of P. andina was similar but slightly smaller, on average, than that of our sample of P. infestans isolates. This is consistent with P. andina being a homoploid hybrid rather than allopolyploid hybrid. Nuclear DNA content was more variable among a smaller sample of P. infestans isolates, including a putative triploid isolate from Mexico, but small differences in nuclear DNA content were also observed among P. andina isolates. Both species appear to be able to tolerate significant variation in genome size. © 2016 by The Mycological Society of America.

  1. Nuclear DNA content of the pigeon orchid (Dendrobium crumenatum Sw. with the analysis of flow cytometry

    Directory of Open Access Journals (Sweden)

    Upatham Meesawat

    2008-05-01

    Full Text Available Nuclear DNA content for the adult plants grown in a greenhouse and in vitro young plantlets of the pigeon orchid (Dendrobium crumenatum Sw. was analyzed using flow cytometry. The resulting 2C DNA values ranged from 2.30±0.14 pgto 2.43±0.06 pg. However, nuclear DNA ploidy levels of long-term in vitro plantlets were found to be triploid and tetraploid.These ploidy levels were confirmed by chromosome counting. Tetraploid individuals (2n = 4x = 76 had approximately two times DNA content than diploid (2n = 2x = 38 individuals. This variation may be due to prolonged cultivation and thepresence of exogenous plant growth regulators.

  2. Study on quantitative reliability analysis by multilevel flow models for nuclear power plants

    International Nuclear Information System (INIS)

    Yang Ming; Zhang Zhijian

    2011-01-01

    Multilevel Flow Models (MFM) is a goal-oriented system modeling method. MFM explicitly describes how a system performs the required functions under stated conditions for a stated period of time. This paper presents a novel system reliability analysis method based on MFM (MRA). The proposed method allows describing the system knowledge at different levels of abstraction which makes the reliability model easy for understanding, establishing, modifying and extending. The success probabilities of all main goals and sub-goals can be available by only one-time quantitative analysis. The proposed method is suitable for the system analysis and scheme comparison for complex industrial systems such as nuclear power plants. (authors)

  3. Flow accelerated corrosion and life management of the secondary circuit of the Embalse nuclear power plant

    International Nuclear Information System (INIS)

    Chocron, Mauricio; La Gamma, Ana M.; Fernandez, Narciso; Moyano, Ricardo; Schiersmann, Christian; Ovando, Luis E.; Sainz, Ricardo A.; Keitelman, Alberto

    2003-01-01

    Flow accelerated corrosion is a matter of concern in secondary circuits of nuclear power plants as well as in fossil fired plants. It contributes to the piping wall thinning and to the corrosion products transport to the steam generators. Because it is a generalized corrosion phenomena, could address to extensive failures. In that sense the plants conduct extensive programs of surveillance of piping degradation. Because the problem involves many variables like alloys, water chemistry and hydrodynamics several models have been proposed in the literature. In the present paper the variables have been organized in a spreadsheet which allows the calculation of normalized risk factors. (author)

  4. In-flow technology for the determination of Sr-90 concentrations in nuclear power plants

    International Nuclear Information System (INIS)

    Dissing, E.

    1982-01-01

    Outlines for a future work concerning in-flow surveillance of Sr-90 in nuclear plant process treams have been studied. The many problems involved with the task of on-line Sr-90 determination were approached in two different ways, one applying β-counting of the 64-hour daughter Y-60 with the use of the process stream itself as a Cerenkov scintillator and the other - indirect - using simultaneous measurement of the concentrations of Sr-91 and Sr-92 for the determination of the leakage route for strontium. (Author)

  5. Initialization effects via the nuclear radius on transverse in-plane flow and its disappearance

    Directory of Open Access Journals (Sweden)

    Bansal Rajni

    2014-04-01

    Full Text Available We study the dependence of collective transverse flow and its disappearance on initialization effects via the nuclear radius within the framework of the Isospin-dependent Quantum Molecular Dynamics (IQMD model. We calculate the balance energy using different parametrizations of the radius available in the literature for the reaction of 12C+12C to explain its measured balance energy. A mass-dependent analysis of the balance energy through out the periodic table is also carried out by changing the default liquid drop IQMD radius.

  6. Groundwater flow modeling focused on the Fukushima Daiichi Nuclear Power Plant Site

    International Nuclear Information System (INIS)

    Saegusa, Hiromitsu; Onoe, Hironori; Kohashi, Akio; Watanabe, Masahisa

    2015-01-01

    Fukushima Daiichi nuclear power plant of Tokyo Electric Power Company is facing contaminated water issues in the aftermath of the Great East Japan Earthquake on March 11, 2011. The amount of contaminated water is continuously increasing due to groundwater leakage into the underground part of reactor and turbine buildings. Therefore, it is important to understand the groundwater flow conditions at the site and to predict the impact of countermeasures taken for isolating groundwater from the source of the contamination, i.e. the reactor buildings. Installations, such as of land-side and sea-side impermeable walls have been planned as countermeasures. In this study, groundwater flow modeling has been performed to estimate the response of groundwater flow conditions to the countermeasures. From the modeling, groundwater recharge and discharge areas, major groundwater flow direction, inflow rate into underground part of the buildings, and changes in response to implementation of the countermeasures could be reasonably estimated. The results indicate that the countermeasures will decrease the volume of inflow into the underground part of the buildings. This means that the countermeasures will be effective in reducing the discharge volume of contaminated groundwater to ocean. (author)

  7. Electric capacitance tomography and two-phase flow for the nuclear reactor safety analysis

    International Nuclear Information System (INIS)

    Lee, Jae Young

    2008-01-01

    Recently electric capacitance tomography has been developed to be used in the analysis of two-phase flow. Although its electric field is not focused as the hard ray tomography such as the X-ray or gamma ray, its convenience of easy access to the system and easy maintenance due to no requirement of radiation shielding benefits us in its application in the two-phase flow study, one of important area in the nuclear safety analysis. In the present paper, the practical technologies in the electric capacitance tomography are represented in both parts of hardware and software. In the software part, both forward problem and inverse problem are discussed and the method of regularization. In the hardware part, the brief discussion of the electronics circuits is made which provides femto farad resolution with a reasonable speed (150 frame/sec for 16 electrodes). Some representative ideal cases are studied to demonstrate its potential capability for the two-phase flow analysis. Also, some variations of the tomography such as axial tomography, and three dimensional tomography are discussed. It was found that the present ECT is expected to become a useful tool to understand the complicated three dimensional two-phase flow which may be an important feature to be equipped by the safety analysis codes. (author)

  8. Determination of the catalyst circulation rate in a FCC cold flow pilot unit using nuclear techniques

    International Nuclear Information System (INIS)

    Santos, Valdemir A. dos; Lima, Emerson A.O.

    2013-01-01

    Nuclear techniques of gamma transmission and radioactive tracer were used to estimate the catalyst circulation rate in a cold flow pilot plant unit of Fluid Catalytic Cracking (FCC). Catalyst circulation rate in a FCC unit, allow to determine operating conditions of the exchange catalyst and inlet data for fluid dynamic simulation computational program. The pilot unit was fabricated obeying geometrical parameters provided by the Petrobras Research Center (CENPES), based on hot pilot units to existing in that center. The cold flow pilot unit has a transfer line, two separation vessels flash type, a return column, a riser and a regenerator. The vertical sections as riser, return column, regenerator column and transfer line are made of transparent material (glass). The two separation vessels have bases with tapered cylindrical shapes and are made of steel plates. The riser is divided into four sections of different diameters (0.005 m, 0.010 m, 0.018 m and 0.025 m) and rising upwards, to simulate the increasing flow rate caused by the increase of volume with the increase of the number of moles due to molecules breakage. The radioactive tracer used was the catalyst itself (intrinsic tracer) irradiated by neutron activation, yielding the radioisotope 59 Fe. The velocity measurements were also obtained with aid of an electronic clock triggered by certain radiation levels across the two detectors. Besides estimates for the catalyst circulation rate was possible to identify the type of flow relative to the catalyst in return column. (author)

  9. Numerical simulation of water flow through the bottom end piece of a nuclear fuel assembly

    International Nuclear Information System (INIS)

    Navarro, Moyses Alberto; Santos, Andre A.C. dos

    2007-01-01

    An experimental and numerical study was conducted on the pressure loss of flows through the bottom end piece of a nuclear fuel assembly. To determine an optimized numerical methodology using the commercial CFD code, CFX 10.0, a series of preliminary simulations of water flows through perforated plates in a square ducts were performed. A perforated plate is a predominant geometry of the bottom end piece, responsible for the majority of the flow's pressure drop. The numerical pressure loss applying an optimized mesh and the k-ε turbulence model showed good agreement when compared with a conventional methodology (Idelchik). Numerical results for the standard bottom end piece were obtained applying the previously determined mesh criteria and the k-ε turbulence model with some geometric simplifications. The agreement between the numerical simulations and experimental results can be considered satisfactory but suggest further numerical investigations with the bottom piece under real conditions of the experiment, without the geometric simplifications and with a gap between the piece and the wall of the flow channel. Additionally, other turbulence models should be appraised for this complex geometry. (author)

  10. Numerical Simulation of Water Flow through the Bottom End Piece of a Nuclear Fuel Assembly

    Science.gov (United States)

    Navarro, Moysés A.; Santos, André A. C. Dos

    An experimental and numerical study was conducted on the pressure loss of flows through the bottom end piece of a nuclear fuel assembly. To determine an optimized numerical methodology using the commercial CFD code, CFX 10.0, a series of preliminary simulations of water flows through perforated plates in a square ducts were performed. A perforated plate is a predominant geometry of the bottom end piece, responsible for the majority of the flow's pressure drop. The numerical pressure loss applying an optimized mesh and the k-ɛ turbulence model showed good agreement when compared with a conventional methodology (Idelchik). Numerical results for the standard bottom end piece were obtained applying the previously determined mesh criteria and the k-ɛ turbulence model with some geometric simplifications. The agreement between the numerical simulations and experimental results can be considered satisfactory but suggests further numerical investigations with the bottom piece under real conditions of the experiment, without the geometric simplifications and with a gap between the piece and the wall of the flow channel. Additionally, other turbulence models should be appraised for this complex geometry.

  11. Implications of competitive markets for operation and funds flow at nuclear plants

    International Nuclear Information System (INIS)

    Bright, R.N.; Leigh, R.W.; Mubayi, V.

    1997-01-01

    The authors employ a detailed electric utility production costing and capacity planning model to examine the use of nuclear power plants under the current (utility-owned) arrangements and under situations which may arise in a competitive market for electric service. The modeling is carried out for a synthetic but realistic power pool based on components of actual utilities in the northeast United States. Under the current arrangements, the costs of nuclear power plant operation are subsumed under capital costs and entered into the open-quotes raw base,close quotes the totality of which determines customer charges. A future competitive environment may be characterized by the absence of a open-quotes rate base,close quotes at least for generation. In this environment, all generation units are in effect independently owned. The authors examine this possibility by estimating the revenue which would flow to nuclear plants at competitive market prices and compare that revenue, to anticipated expenses for fuel, O ampersand M and amortization of capital (where appropriate), and for potential additional expenses such as safety requirements and payments to decommissioning funds. They also incorporate financial relief for the portion of these costs which can be considered open-quotes stranded investmentsclose quotes based on FERC and prospective state regulatory policies. In this competitive environment some nuclear plants may not remain economically viable. Their replacement by newly constructed fossil-fueled units will require substantial capital and lead to increased emissions and fuel use, which are calculated. The authors calculations show that while it may be clearly economically advantageous for individual owners to shut down unviable nuclear capacity, such early retirements may result in significant economic losses to the power pool as a whole

  12. Multiphase, multicomponent flow and transport models for Nuclear Test-Ban Treaty monitoring and nuclear waste disposal applications

    Science.gov (United States)

    Jordan, Amy

    Open challenges remain in using numerical models of subsurface flow and transport systems to make useful predictions related to nuclear waste storage and nonproliferation. The work presented here addresses the sensitivity of model results to unknown parameters, states, and processes, particularly uncertainties related to incorporating previously unrepresented processes (e.g., explosion-induced fracturing, hydrous mineral dehydration) into a subsurface flow and transport numerical simulator. The Finite Element Heat and Mass (FEHM) transfer code is used for all numerical models in this research. An experimental campaign intended to validate the predictive capability of numerical models that include the strongly coupled thermal, hydrological, and chemical processes in bedded salt is also presented. Underground nuclear explosions (UNEs) produce radionuclide gases that may seep to the surface over weeks to months. The estimated timing of gas arrival at the surface may be used to deploy personnel and equipment to the site of a suspected UNE, if allowed under the terms of the Comprehensive Nuclear Test-Ban Treaty. A model was developed using FEHM that considers barometrically pumped gas transport through a simplified fractured medium and was used to quantify the impact of uncertainties in hydrologic parameters (fracture aperture, matrix permeability, porosity, and saturation) and season of detonation on the timing of gas breakthrough. Numerical sensitivity analyses were performed for the case of a 1 kt UNE at a 400 m burial depth. Gas arrival time was found to be most affected by matrix permeability and fracture aperture. Gases having higher diffusivity were more sensitive to uncertainty in the rock properties. The effect of seasonality in the barometric pressure forcing was found to be important, with detonations in March the least likely to be detectable based on barometric data for Rainier Mesa, Nevada. Monte Carlo modeling was also used to predict the window of

  13. Nuclear matter flow in the Kr+Au collisions at 43 MeV/u

    International Nuclear Information System (INIS)

    Bougault, R.; Delaunay, F.; Genoux-Lubain, A.; Lebrun, C.; Lecolley, J.F.; Lefebvres, F.; Louvel, M.; Steckmeyer, J.C.; Aloff, J.C.; Bilwes, B.; Bilwes, R.; Glaser, M.; Rudolf, G.; Scheibling, F.; Stuttge, L.

    1989-01-01

    When heavy nuclei collide at energy far above the Coulomb barrier we may study the property of nuclear matter in temperature and also density regions far away from the equilibrium. We then hope to study dynamical effects related to the compressibility and the two body collision term. At relativistic energies, some collective effects (flow, bounce off) have been established from a shape analysis done with a large number of light particles with Z ≤ 10. For incident energies lower than 100 MeV/u we may expect that the number of nuclear species formed will be smaller and that a large part of the nuclear matter involved in the collision will be shared in a limited number of heavy fragments (Z ≥ 10). If dynamical effects are still present at GANIL energies they ought to manifest themselves through the properties of the produced fragments (masses, emission angles, velocities and correlated variables). We will present an analysis of heavy nuclei collisions at 43 MeV/u based on as exclusive as possible detection of large fragments

  14. Dispersed Two-Phase Flow Modelling for Nuclear Safety in the NEPTUNE_CFD Code

    Directory of Open Access Journals (Sweden)

    Stephane Mimouni

    2017-01-01

    Full Text Available The objective of this paper is to give an overview of the capabilities of Eulerian bifluid approach to meet the needs of studies for nuclear safety regarding hydrogen risk, boiling crisis, and pipes and valves maintenance. The Eulerian bifluid approach has been implemented in a CFD code named NEPTUNE_CFD. NEPTUNE_CFD is a three-dimensional multifluid code developed especially for nuclear reactor applications by EDF, CEA, AREVA, and IRSN. The first set of models is dedicated to wall vapor condensation and spray modelling. Moreover, boiling crisis remains a major limiting phenomenon for the analysis of operation and safety of both nuclear reactors and conventional thermal power systems. The paper aims at presenting the generalization of the previous DNB model and its validation against 1500 validation cases. The modelling and the numerical simulation of cavitation phenomena are of relevant interest in many industrial applications, especially regarding pipes and valves maintenance where cavitating flows are responsible for harmful acoustics effects. In the last section, models are validated against experimental data of pressure profiles and void fraction visualisations obtained downstream of an orifice with the EPOCA facility (EDF R&D. Finally, a multifield approach is presented as an efficient tool to run all models together.

  15. Numerical Simulation of Three-Dimensional Flow Through Full Passage and Performance Prediction of Nuclear Reactor Coolant Pump

    International Nuclear Information System (INIS)

    Li Ying; Zhou Wenxia; Zhang Jige; Wang Dezhong

    2009-01-01

    In order to achieve the level of self-design and domestic manufacture of the reactor coolant pump (nuclear main pump), the software FLUENT was used to simulate the three-dimensional flow through full passage of one nuclear main pump basing on RNG κ-ε turbulence model and SIMPLE algorithm. The distribution of pressure and velocity of the flow in the impeller's surface was analyzed in different working conditions. Moreover, the performance of the pump was predicted based on the simulation results. The results show that the distributions of pressure and velocity are reasonable in both the working and back face of the blade in the steady working condition. The pressure of the flow is increased from the inlet to the outlet of the pump, and shows the maximal value in the impeller region. Comparatively satisfactory efficiency and head value were obtained in the condition of the pump design. The shaft power of the nuclear main pump is gradually increased with the increase of the flow flux. These results are helpful in understanding the change of the internal flow field in the nuclear main pump, which is of some importance for the pre-exploration and theoretical research on the domestic manufacture of the nuclear main pump. (authors)

  16. Experimental investigations on turbulent mixing of hot upward flow and cold downward flow inside a chimney model of a nuclear reactor

    Energy Technology Data Exchange (ETDEWEB)

    Sengupta, Samiran, E-mail: samiran_sengupta@yahoo.co.in [Research Reactor Design & Projects Division, Bhabha Atomic Research Centre, Mumbai 400085 (India); Ghosh, Aniruddha [Research Reactor Design & Projects Division, Bhabha Atomic Research Centre, Mumbai 400085 (India); Sengupta, C. [Research Reactor Maintenance Division, Bhabha Atomic Research Centre, Mumbai 400085 (India); Vijayan, P.K. [Reactor Design & Development Group, Bhabha Atomic Research Centre, Mumbai 400085 (India); Bhattacharya, S. [Research Reactor Design & Projects Division, Bhabha Atomic Research Centre, Mumbai 400085 (India); Sharma, R.C. [Reactor Group, Bhabha Atomic Research Centre, Mumbai 400085 (India)

    2016-02-15

    Highlights: • Simulated mixing of hot upward and cold downward flows in a chimney of a reactor. • Experiments in chimney model (2:9 scale) at Reynolds number (Re)—1.5 to 4.5 × 10{sup 5}. • Hot upward flow comes out of the chimney when bypass flow ratio (R) is zero. • Increase in ratio (R) reduces jet height, vortex spread height and temperature front height. • Effects of Re, chimney height and temperature differential are not significant. - Abstract: Experiments were conducted to study the turbulent mixing of hot upward flow and cold downward flow inside a scaled down model of chimney structure of a pool type nuclear research reactor. Open pool type nuclear reactors often use this type of chimney structures to prevent mixing of radioactive core outlet water directly into the reactor pool so that radiation field at the reactor pool top can be kept to a lower limit. The chimney structure is designed to facilitate guiding of the radioactive water towards the two outlet nozzles of the chimney and simultaneously allows drawing water from the reactor pool through the chimney top opening. The present work aims at studying flow mixing behaviour of hot and cold water inside a 2/9th scaled down model of the chimney structure experimentally. The ratio between the cold downward flow and the hot upward flow is varied between 0 and 0.15 to predict the extent of suppression of the hot upward flow within the chimney region for various bypass flow ratios. The Reynolds number of the hot upward flow considered in the experiment is about 1.5 × 10{sup 5} which corresponds to a flow rate of about 500 l min{sup −1}. The upward jet height and the temperature distribution were predicted from the experiment. It was observed that increase in bypass flow ratio reduces the upward jet height of hot water. Experiments were also carried out by increasing the flow rate to 1000 and 1500 l min{sup −1} corresponding to Reynolds numbers of 3 × 10{sup 5} and 4.5 × 10{sup 5

  17. Modular assembly for supporting, straining, and directing flow to a core in a nuclear reactor

    Science.gov (United States)

    Pennell, William E.

    1977-01-01

    A reactor core support arrangement for supporting, straining, and providing fluid flow to the core and periphery of a nuclear reactor during normal operation. A plurality of removable inlet modular units are contained within permanent liners in the lower supporting plate of the reactor vessel lower internals. During normal operation (1) each inlet modular unit directs main coolant flow to a plurality of core assemblies, the latter being removably supported in receptacles in the upper portion of the modular unit and (2) each inlet modular unit may direct bypass flow to a low pressure annular region of the reactor vessel. Each inlet modular unit may include special fluid seals interposed between mating surfaces of the inlet modular units and the core assemblies and between the inlet modular units and the liners, to minimize leakage and achieve an hydraulic balance. Utilizing the hydraulic balance, the modular units are held in the liners and the assemblies are held in the modular unit receptacles by their own respective weight. Included as part of the permanent liners below the horizontal support plate are generally hexagonal axial debris barriers. The axial debris barriers collectively form a bottom boundary of a secondary high pressure plenum, the upper boundary of which is the bottom surface of the horizontal support plate. Peripheral liners include radial debris barriers which collectively form a barrier against debris entry radially. During normal operation primary coolant inlet openings in the liner, below the axial debris barriers, pass a large amount of coolant into the inlet modular units, and secondary coolant inlet openings in the portion of the liners within the secondary plenum pass a small amount of coolant into the inlet modular units. The secondary coolant inlet openings also provide alternative coolant inlet flow paths in the unlikely event of blockage of the primary inlet openings. The primary inlet openings have characteristics which limit the

  18. CFD analysis of moderator flow and temperature fields inside a vertical calandria vessel of nuclear reactor

    Energy Technology Data Exchange (ETDEWEB)

    Kansal, Anuj Kumar, E-mail: akansal@barc.gov.in [Bhabha Atomic Research Centre, Trombay, Mumbai 400085 (India); Joshi, Jyeshtharaj B., E-mail: jbjoshi@gmail.com [Homi Bhabha National Institute, Anushaktinagar, Mumbai 400094 (India); Maheshwari, Naresh Kumar, E-mail: nmahesh@barc.gov.in [Bhabha Atomic Research Centre, Trombay, Mumbai 400085 (India); Vijayan, Pallippattu Krishnan, E-mail: vijayanp@barc.gov.in [Bhabha Atomic Research Centre, Trombay, Mumbai 400085 (India)

    2015-06-15

    Highlights: • 3D CFD of vertical calandria vessel. • Spatial distribution of volumetric heat generation. • Effect of Archimedes number. • Non-dimensional analysis. - Abstract: Three dimensional computational fluid dynamics (CFD) analysis has been performed for the moderator flow and temperature fields inside a vertical calandria vessel of nuclear reactor under normal operating condition using OpenFOAM CFD code. OpenFOAM is validated by comparing the predicted results with the experimental data available in literature. CFD model includes the calandria vessel, calandria tubes, inlet header and outlet header. Analysis has been performed for the cases of uniform and spatial distribution of volumetric heat generation. Studies show that the maximum temperature in moderator is lower in the case of spatial distribution of heat generation as compared to that in the uniform heat generation in calandria. In addition, the effect of Archimedes number on maximum and average moderator temperature was investigated.

  19. Numerical simulation of water flow through the bottom en piece of a nuclear fuel assembly

    International Nuclear Information System (INIS)

    Navarro, Moyses A.; Santos, Andre A. Campagnole dos

    2007-01-01

    The water flow through the bottom nozzle of a nuclear fuel assembly was simulated using a commercial CFD code, CFX 10.0. Previously, simulations with a perforated plate similar to the bottom nozzle plate were performed to define the appropriate mesh refinement and turbulence model (κ-ε or SST). Subsequently, the numerical simulation was performed with the optimized mesh using the turbulence model (κ-ε in a standard bottom nozzle with some geometric simplifications. The numerical results were compared with experimental results to determine the pressure drop through the bottom nozzle in the Reynolds range from ∼10500 to ∼95000. The agreement between the numerical simulations and experimental results may be considered satisfactory. The study indicated that the CFD codes can play an important role in the development of pieces with complex geometries, optimizing the planning of the experiments and aiding in the experimental analysis. (author)

  20. Polyploidy in the olive complex (Olea europaea): Evidence from flow cytometry and nuclear microsatellite analyses

    DEFF Research Database (Denmark)

    Besnard, G.; Garcia-Verdugo, C.; Rubio de Casas, R.

    2008-01-01

    Background: Phylogenetic and phylogeographic investigations have been previously performed to study the evolution of the olive tree complex (Olea europaea). A particularly high genomic diversity has been found in north-west Africa. However, to date no exhaustive study has been addressed to infer...... putative polyploidization events and their evolutionary significance in the diversification of the olive tree and its relatives. Methods: Representatives of the six olive subspecies were investigated using (a) flow cytometry to estimate genome content, and (b) six highly variable nuclear microsatellites....... Lastly, abnormalities in chromosomes inheritance leading to aneuploid formation were revealed using microsatellite analyses in the offspring from the controlled cross in subsp. maroccana. Conclusions: This study constitutes the first report for multiple polyploidy in olive tree relatives. Formation...

  1. Study of visualized analysis platform for nuclear fuel cycle system based on multilevel flow model

    International Nuclear Information System (INIS)

    Liu Jingquan

    2005-01-01

    Complex Nuclear Fuel Cycle (NFC) system faces many socio-technical issues that need to obtain the consensus between stake holders of different knowledge background. In this paper, a visualized analysis platform based on graphical functional modeling method, Multilevel Flow Model (MFM), was proposed to help those stake holders to recognize and analyze various socio-technical issues in NFC system. There are some new functions, such as 'Reaction Function', 'Switch Function' and 'Conversion Function', introduced to adapt new simulation tasks for NFC system. Based upon this methodology, a micro-process and a macro-process of NFC system were simulated and meanwhile some key analysis variables required by some analysis methods were deducted and displayed in the platform. And finally a simple simulation analysis was conducted based on the proposed MFM application. (author)

  2. Development and comparision of techniques for estimating design basis flood flows for nuclear power plants

    International Nuclear Information System (INIS)

    1980-05-01

    Estimation of the design basis flood for Nuclear Power Plants can be carried out using either deterministic or stochastic techniques. Stochastic techniques, while widely used for the solution of a variety of hydrological and other problems, have not been used to date (1980) in connection with the estimation of design basis flood for NPP siting. This study compares the two techniques against one specific river site (Galt on the Grand River, Ontario). The study concludes that both techniques lead to comparable results , but that stochastic techniques have the advantage of extracting maximum information from available data and presenting the results (flood flow) as a continuous function of probability together with estimation of confidence limits. (author)

  3. Effects of air flow rate on the oxidation of NBG-18 and NBG-25 nuclear graphite

    Science.gov (United States)

    Chi, Se-Hwan; Chan Kim, Gen

    2017-08-01

    The effects of air flow rate (FR) (FR range: 1-10 L/min) on the oxidation of NBG-18 and NBG-25 nuclear graphite grades at temperatures between 600 and 1100 °C were studied, in reference to the standard test procedure for measuring oxidation rates of nuclear graphite in air (ASTM D 7542-09). The results showed that the FR effects on oxidation rate (OR) increase with increasing temperature with negligible FR effects at 600 °C for both materials. At high temperatures (>800 °C) there appears to be a two-stage relationship between FR and OR, which corresponds to the transition between reaction rates dominated by chemical kinetics and those dominated by diffusion. The material-specific microstructure appeared strongly influences this transition. The overall OR-FR behaviours of NBG-18 were higher than NBG-25 at 600-800 °C while negligible differences in the OR-FR behaviours between the two grades were observed at 900-1100 °C. The mercury porosimetry data showed that the higher OR-FR behaviours observed in NBG-18 may partly be attributed to the differences in the pore size distribution (open porosity and cumulative pore area) between the grades, especially for the large size pores (diameter ≫ 5 × 103 nm).

  4. Improved and Reproducible Flow Cytometry Methodology for Nuclei Isolation from Single Root Meristem

    Directory of Open Access Journals (Sweden)

    Thaís Cristina Ribeiro Silva

    2010-01-01

    Full Text Available Root meristems have increasingly been target of cell cycle studies by flow cytometric DNA content quantification. Moreover, roots can be an alternative source of nuclear suspension when leaves become unfeasible and for chromosome analysis and sorting. In the present paper, a protocol for intact nuclei isolation from a single root meristem was developed. This proceeding was based on excision of the meristematic region using a prototypical slide, followed by short enzymatic digestion and mechanical isolation of nuclei during homogenization with a hand mixer. Such parameters were optimized for reaching better results. Satisfactory nuclei amounts were extracted and analyzed by flow cytometry, producing histograms with reduced background noise and CVs between 3.2 and 4.1%. This improved and reproducible technique was shown to be rapid, inexpensive, and simple for nuclear extraction from a single root tip, and can be adapted for other plants and purposes.

  5. Identification of two-phase flow patterns in a nuclear reactor by high-frequency contribution fraction

    International Nuclear Information System (INIS)

    Wang, Y.W.; Pei, B.S.

    1990-01-01

    A method based on noise analysis techniques that can be applied to the identification of two-phase flow patterns in nuclear reactors is proposed. The identifying criterion, the high-frequency contribution fraction (HFCF), offers new potential to the in-core recognition of two-phase flow patterns. By analyzing 76 sets of signals acquired from a research nuclear reactor where two-phase flow patterns are generated in an in-core air/water loop, the typical signal, autocorrelogram, and spectrum of each flow pattern are demonstrated and evaluated. The identification success rate is 87 or 93%, depending on whether churn flow is counted. A method to improve the identification rate is also presented. This study demonstrates that the fluctuation characteristics above 10 Hz are induced by two-phase flow itself and are independent of the driving source; thus, it is adequate to apply the HFCF to the identification of two-phase flow patters. This study shows that it is possible to identify two-phase flow patterns by HFCF values

  6. Peak expiratory flow rate (PEFR) among Nuclear Fuel Complex (NFC) employees

    International Nuclear Information System (INIS)

    Vijay Rao, J.; Venkaiah, K.; Mohan Rao, N.

    2010-01-01

    At Nuclear Fuel Complex (NFC), employees are exposed to ammonia, hydrofluoric acid, acetone, etc., which are respiratory toxicants and inhalation of these pollutants may produce irritation and obstruction in airways. Due to nature of their occupation, tradesman working in plants are having longer duration of exposure (LDE) and others, such as supervisors, scientific officers, helpers, etc., that occasionally visit plants are having shorter duration of exposure (SDE) to these pollutants. The peak expiratory flow rate (PEFR) is an index to diagnose obstruction in larger airways and this is metered with mini peak flow meter among 835 NFC employees. Using ANOVA test, PEFR value was compared according to age and smoking. The value was compared between LDE and SDE employees according to smoking and duration of employment. The multiple regression equation for prediction of PEFR was developed. Age, smokers and higher duration of LDE employees demonstrated significantly lower PEFR value. In comparison to 10 year duration, 30 and above year duration of employment, LDE employees showed a higher decline in PEFR, that is 95 L (17.6%) and in SDE employees, that is 41L (7.8%). This may be due to longer duration of employment of LD employees smoking prevention and follow up study is suggested. (author)

  7. Groundwater and vadose Zone Integration Project Nuclear Material Mass Flow and Accountability on the Hanford Site

    International Nuclear Information System (INIS)

    GRASHER, A.A.

    2001-01-01

    The purpose of this report is to provide a discussion of the accountable inventory of Hanford Site nuclear material (NM) over the operating period. This report does not provide judgments on impacts to the Hanford Site environs by the reported waste streams or inventory. The focus of this report is on the processes, facilities, and process streams that constituted the flow primarily of plutonium and uranium through the Hanford Site. The material balance reports (MBRS) are the basis of the NM accountable inventory maintained by each of the various contractors used by the U.S. Department of Energy (DOE) and its predecessors to operate the Hanford Site. The inventory was tracked in terms of a starting inventory, receipts, transfers, and ending inventory. The various components of the inventory are discussed as well as the uncertainty in the measurement values used to establish plant inventory and material transfers. The accountable NM inventory does not report all the NM on the Hanford Site and this difference is discussed relative to some representative nuclides. The composition and location of the current accountable inventory are provided, as well as the latest approved set (2000) of flow diagrams of the proposed disposition of the excess accountable NM inventory listed on the Idaho National Engineering and Environmental Laboratory (INEEL) web page

  8. Information flow among researchers at IPEN - Instituto de Pesquisas Energeticas e Nucleares

    International Nuclear Information System (INIS)

    Sugai, Mioka

    1986-01-01

    The information flow among a group of researchers at the Instituto de Pesquisas Energeticas e Nucleares of the Comissao Nacional de Energia Nuclear/Sao Paulo (IPEN-CNEN/SP) was analysed by means of a study of use and non-use of formal and informal information channels. The study proposed suggesting ideas for the improvement of the information network, as a means of contributing to the future planning of the information transfer structure among the I PEN technical-scientific community. A structural interview was used to collect the data. The researchers were characterized under functional, academic and professional aspects. Their information needs were identified as well as the factors which affect such needs. The researchers behaviour while searching for information was analysed by means of the critical incident technique. The informal communication networks were also identified according to a sociometric technique. The results show that in the Department included in the study, information flows equally through formal and informal channels. It is evident that there is a small correlation between degree of use and degree of importance of information sources. There is no evidence that those who make little use of formal channels supply their information needs by use of informal channels. It was patent that non-accessibility is the key factor which influences the non-use of information. The motivation of the use of formal sources is significantly inhibited by the fact that the library collection is not brought up-to-date. Relatively intense informal communication was verified both inter and intra-divisions. It is also evident that researchers with higher academic degree make more frequent use of formal channels, and stand a greater possibility of being identified as gatekeepers. However, those researchers who are considered more productive, at their present function, are not always those who make more frequent use of formal channels. The conclusions show that in order

  9. Flow accelerated corrosion programme in Angra nuclear power plants Units 1 and 2

    International Nuclear Information System (INIS)

    Costa, Tomas D.S.

    2009-01-01

    Almirante Alvaro Alberto Nuclear Power Plant, referred herein as Angra NPP, comprises two generating units operated by ELETROBRAS Termonuclear S.A. - ELETRONUCLEAR. Unit 1, a Westinghouse designed PWR, operates commercially since 1982 generating 657 MW, while unit 2, designed by Siemens, is a 1350 MW PWR in operation since 2000. Flow Accelerated Corrosion (FAC) programme integrate a set of ageing related programmes established to control ageing effects on equipment and piping in order to ensure their capability to perform their functions during the plant remaining life. The objective of this paper is to describe ELETRONUCLEAR experience in developing and implementing a programme related to Flow Accelerated Corrosion, with focus on piping of the secondary sides of Angra NPP, explaining in detail the current situation for important pipings, and presenting updated results of the FAC control. Accurate inspections are the basis for an effective FAC programme. Wall thickness measurements establish the extent of wear in a given piping, providing data to support FAC trends, and to refine the predictive model. A permanently and clearly identify grid is makes re-inspection possible, at any time, at the same points. Supported by the computational system Comsy, some of its features as FAC potentially susceptible lines identification, wall thinning rates determination, inspection data recording, wear rates analyses, lifetime prediction and inspection scheduling will be showed in detail. This programme has started in 2004, and since this time it was applied in a total of six outages in Angra 1 NPP and three outages in Angra 2 NPP, with a large number of piping elements inspected and analyzed. Some of these results will be showed and discussed in this paper, regarding the potential of Flow Accelerated Corrosion occurrence. (author)

  10. Development of improved thermal hydraulics and fuel performance technology; development of turbulence model and simulation code for flow analysis in nuclear fuel assembly

    Energy Technology Data Exchange (ETDEWEB)

    Myung, H. K.; Yang, S. Y.; Kim, B. H.; Song, J. H.; Oh, J. Z. [Kookmin University, Seoul (Korea)

    2002-03-01

    The flow through a nuclear rod bundle with mixing vanes is very complex and so required a suitable turbulence model for its accurate prediction. Subchannel flow in a nuclear bundle having vanes to mix flow appears complex turbulent flow. Objective of this study is to investigate performance of prediction about turbulence model contained in STAR-CD code and to develop suitable turbulence model which can predict complex flow in nuclear assembly. For several nonlinear {kappa}-{epsilon} turbulence models, their performance were investigated in the prediction of the flow in nuclear fuel assembly, and also their problems were discussed in detail. The results obtained from the present research would give a help for the development of turbulence model which can accurately predict the flow through the rod bundles with mixing vanes. 19 refs., 32 figs., 3 tabs. (Author)

  11. Control flow of radiopharmaceuticals in nuclear medicine by means of an E-service; Control flujo de radiofarmacos en medicine nuclear por medio de un E-servicio

    Energy Technology Data Exchange (ETDEWEB)

    Nunez Martin, L.; Gonzalez de Mingo, M. A.; Fragua Redondo, J. A.; Martinez Ortega, J.; Gutierrez Camunas, S.; Redondo Miguel, A. B.

    2013-07-01

    The almost generalized use of single-dose Nuclear Medicine for performing diagnostic tests or treatments, and the consequent complexity that accompanies its management, has resulted in the need to control the flow of material radioisotopic tools. An e-service is designed to manage the flow of radiopharmaceuticals and control its use and spending. This control does not only affect the efficiency in the use and cost of material, but in the radioactive waste associated with the non-use and waste reduction and a more effective organization of the Department. (Author)

  12. Nuclear energy in Europe: uranium flow modeling and fuel cycle scenario trade-offs from a sustainability perspective.

    Science.gov (United States)

    Tendall, Danielle M; Binder, Claudia R

    2011-03-15

    The European nuclear fuel cycle (covering the EU-27, Switzerland and Ukraine) was modeled using material flow analysis (MFA).The analysis was based on publicly available data from nuclear energy agencies and industries, national trade offices, and nongovernmental organizations. Military uranium was not considered due to lack of accessible data. Nuclear fuel cycle scenarios varying spent fuel reprocessing, depleted uranium re-enrichment, enrichment assays, and use of fast neutron reactors, were established. They were then assessed according to environmental, economic and social criteria such as resource depletion, waste production, chemical and radiation emissions, costs, and proliferation risks. The most preferable scenario in the short term is a combination of reduced tails assay and enrichment grade, allowing a 17.9% reduction of uranium demand without significantly increasing environmental, economic, or social risks. In the long term, fast reactors could theoretically achieve a 99.4% decrease in uranium demand and nuclear waste production. However, this involves important costs and proliferation risks. Increasing material efficiency is not systematically correlated with the reduction of other risks. This suggests that an overall optimization of the nuclear fuel cycle is difficult to obtain. Therefore, criteria must be weighted according to stakeholder interests in order to determine the most sustainable solution. This paper models the flows of uranium and associated materials in Europe, and provides a decision support tool for identifying the trade-offs of the alternative nuclear fuel cycles considered.

  13. Effect of water chemistry improvement on flow accelerated corrosion in light-water nuclear reactor

    International Nuclear Information System (INIS)

    Sugino, Wataru; Ohira, Taku; Nagata, Nobuaki; Abe, Ayumi; Takiguchi, Hideki

    2009-01-01

    Flow Accelerated Corrosion (FAC) of Carbon Steel (CS) piping has been one of main issues in Light-Water Nuclear Reactor (LWRs). Wall thinning of CS piping due to FAC increases potential risk of pipe rupture and cost for inspection and replacement of damaged pipes. In particular, corrosion products generated by FAC of CS piping brought steam generator (SG) tube corrosion and degradation of thermal performance, when it intruded and accumulated in secondary side of PWR. To preserve SG integrity by suppressing the corrosion of CS, High-AVT chemistry (Feedwater pH9.8±0.2) has been adopted to Tsuruga-2 (1160 MWe PWR, commercial operation in 1987) in July 2005 instead of conventional Low-AVT chemistry (Feedwater pH 9.3). By the High-AVT adoption, the accumulation rate of iron in SG was reduced to one-quarter of that under conventional Low-AVT. As a result, a tendency to degradation of the SG thermal efficiency was improved. On the other hand, it was clarified that High-AVT is ineffective against Flow Accelerated Corrosion (FAC) at the region where the flow turbulence is much larger. By contrast, wall thinning of CS feed water pipes due to FAC has been successfully controlled by oxygen treatment (OT) for long time in BWRs. Because Magnetite film formed on CS surface under AVT chemistry has higher solubility and porosity in comparison with Hematite film, which is formed under OT. In this paper, behavior of the FAC under various pH and dissolved oxygen concentration are discussed based on the actual wall thinning rate of BWR and PWR plant and experimental results by FAC test-loop. And, it is clarified that the FAC is suppressed even under extremely low DO concentration such as 2ppb under AVT condition in PWR. Based on this result, we propose the oxygenated water chemistry (OWC) for PWR secondary system which can mitigate the FAC of CS piping without any adverse effect for the SG integrity. Furthermore, the applicability and effectiveness of this concept developed for FAC

  14. Application of flow network models of SINDA/FLUINT{sup TM} to a nuclear power plant system thermal hydraulic code

    Energy Technology Data Exchange (ETDEWEB)

    Chung, Ji Bum [Institute for Advanced Engineering, Yongin (Korea, Republic of); Park, Jong Woon [Korea Electric Power Research Institute, Taejon (Korea, Republic of)

    1998-12-31

    In order to enhance the dynamic and interactive simulation capability of a system thermal hydraulic code for nuclear power plant, applicability of flow network models in SINDA/FLUINT{sup TM} has been tested by modeling feedwater system and coupling to DSNP which is one of a system thermal hydraulic simulation code for a pressurized heavy water reactor. The feedwater system is selected since it is one of the most important balance of plant systems with a potential to greatly affect the behavior of nuclear steam supply system. The flow network model of this feedwater system consists of condenser, condensate pumps, low and high pressure heaters, deaerator, feedwater pumps, and control valves. This complicated flow network is modeled and coupled to DSNP and it is tested for several normal and abnormal transient conditions such turbine load maneuvering, turbine trip, and loss of class IV power. The results show reasonable behavior of the coupled code and also gives a good dynamic and interactive simulation capabilities for the several mild transient conditions. It has been found that coupling system thermal hydraulic code with a flow network code is a proper way of upgrading simulation capability of DSNP to mature nuclear plant analyzer (NPA). 5 refs., 10 figs. (Author)

  15. Development of a mobile dynamic nuclear polarizer for continuous flow applications

    Energy Technology Data Exchange (ETDEWEB)

    Ebert, Sandro; Dollmann, Bjoern; Bauer, Christian; Koelzer, Michael; Spiess, Hans W.; Hinderberger, Dariush; Muennemann, Kerstin [Max Planck Institute for Polymer Research, Mainz (Germany); Bluemler, Peter [Johannes Gutenberg University, Institute of Physics, Mainz (Germany)

    2011-07-01

    Despite its wide applicability in natural science, NMR still suffers from its inherently low sensitivity. This could be overcome by hyperpolarization of molecules via dynamic nuclear polarization (DNP). Here, we introduce a mobile DNP polarizer, based on an inexpensive Halbach magnet operating at 0.35 T. It shows an almost vanishing magnetic flux at its outer side and is not disturbing other instruments. It can be placed directly next to a superconducting magnet, thus limiting the transport time of the hyperpolarized sample. It will be shown, that the Halbach magnet has the same DNP performance like an electromagnet. Although DNP methods have found important applications in science, two problems remain: Firstly radicals are needed, which are mostly toxic. This problem becomes crucial with regard to medical applications. Secondly, the sample must be transported from the polarization magnet to the place of detection and polarization losses due to T1 occur. We are currently implementing a flow system to the mobile DNP polarizer, which should overcome both obstacles. The radicals will be immobilized in a gel matrix and the hyperpolarized radical free fluid is pumped subsequently directly in the MRI scanner.

  16. Possibilities and Limitations of CFD Simulation for Flashing Flow Scenarios in Nuclear Applications

    Directory of Open Access Journals (Sweden)

    Yixiang Liao

    2017-01-01

    Full Text Available The flashing phenomenon is relevant to nuclear safety analysis, for example by a loss of coolant accident and safety release scenarios. It has been studied intensively by means of experiments and simulations with system codes, but computational fluid dynamics (CFD simulation is still at the embryonic stage. Rapid increasing computer speed makes it possible to apply the CFD technology in such complex flow situations. Nevertheless, a thorough evaluation on the limitations and restrictions is still missing, which is however indispensable for reliable application, as well as further development. In the present work, the commonly-used two-fluid model with different mono-disperse assumptions is used to simulate various flashing scenarios. With the help of available experimental data, the results are evaluated, and the limitations are discussed. A poly-disperse method is found necessary for a reliable prediction of mean bubble size and phase distribution. The first attempts to trace the evolution of the bubble size distribution by means of poly-disperse simulations are made.

  17. The transition criteria of circulating flow pattern of moderator in the calandria tank of CANDU nuclear power plant

    International Nuclear Information System (INIS)

    Jung, Yun Sik; Lee, Jae Young; Kim, Man Woong

    2004-01-01

    The moderator cooling system to the Calandria tank of CANDU nuclear power plant provides an alternative pass of heat sink during the hypothetical loss of coolant accident. Also, the neutron population in the CANDU plant can be affected by the moderator temperature change which strongly depends on the circulating flow pattern in the Calandria tank. It has been known that there are three distinguished flow patterns: the buoyancy dominated flow, the momentum dominated flow, and the mixed type flow. The Canadian Nuclear Safety Commission (CNSC) recommended that a series of experimental works should be performed to verify the three dimensional codes. Two existing facilities, SPEL (1982) and STERN (1990), have produced experimental data for these purposes. The present work is also motivated to build up a new scaled experimental facility named HGU for the same purposes. CANDU-6 was selected as the target plant to be scaled down. In the design for the scaled facility, the knowledge on the flow regime transitions in the circulating flow was imperative. In the present study, to pave the way for the scaling, the flow pattern maps of circulating flow were constructed based on the Reynolds number and Archimedes number. The CFX code was employed with real meshes to represent all calandria tubes in the tank. The flow pattern maps were constructed for SPEL, STERN, HGU, and CANDU6. As the key transition criterion useful for scaling law, a new Archimedes number considering the jet impingement of the feed water in the Calandria tank was found. The transition of flow patterns was made with the same Archimedes number for CANDU6, STERN and HGU. However, SPEL which has third of the modified Archimedes number showed different maps in the wider region of mixed flow pattern was observed. It was found that the Archimedes number considering the inlet nozzle velocity plays the key role in patterns classification. Also, it can be suggested that the moderator cooling system needs to be designed

  18. On the flow of groundwater in closed tunnels. Generic hydrogeological modelling of nuclear waste repository, SFL 3-5

    International Nuclear Information System (INIS)

    Holmen, J.G.

    1997-06-01

    The purpose is to study the flow of groundwater in closed tunnels by use of mathematical models. The calculations were based on three dimensional models, presuming steady state conditions. The stochastic continuum approach was used for representation of a heterogeneous rock mass. The size of the calculated flow is given as a multiple of an unknown regional groundwater flow. The size of the flow in a tunnel has been studied, as regards: Direction of the regional groundwater flow, Tunnel length, width and conductivity; Heterogeneity of the surrounding rock mass; Flow barriers and encapsulation inside a tunnel. The study includes a model of the planned repository for nuclear waste (SFL 3-5). The flow through the tunnels is estimated for different scenarios. The stochastic continuum approach has been investigated, as regards the representation of a scale dependent heterogeneous conductivity. An analytical method is proposed for the scaling of measured conductivity values, the method is consistent with the stochastic continuum approach. Some general conclusions from the work are: The larger the amount of heterogeneity, the larger the expected flow; The effects of the heterogeneity will decrease with increased tunnel length; If the conductivity of the tunnel is smaller than a threshold value, the tunnel conductivity is the most important parameter; If the tunnel conductivity is large and the tunnel is long, the most important parameter is the direction of the regional flow; Given a heterogeneous rock mass, if the tunnel length is shorter than about 500 m, the heterogeneity will be an important parameter, for lengths shorter than about 250 m, probably the most important; The flow through an encapsulation surrounded by a flow barrier is mainly dependent on the conductivity of the barrier. 70 refs, 110 figs, 10 tabs

  19. On the flow of groundwater in closed tunnels. Generic hydrogeological modelling of nuclear waste repository, SFL 3-5

    Energy Technology Data Exchange (ETDEWEB)

    Holmen, J.G. [Uppsala Univ. (Sweden). Inst. of Earth Sciences]|[Golder Associates AB (Sweden)

    1997-06-01

    The purpose is to study the flow of groundwater in closed tunnels by use of mathematical models. The calculations were based on three dimensional models, presuming steady state conditions. The stochastic continuum approach was used for representation of a heterogeneous rock mass. The size of the calculated flow is given as a multiple of an unknown regional groundwater flow. The size of the flow in a tunnel has been studied, as regards: Direction of the regional groundwater flow, Tunnel length, width and conductivity; Heterogeneity of the surrounding rock mass; Flow barriers and encapsulation inside a tunnel. The study includes a model of the planned repository for nuclear waste (SFL 3-5). The flow through the tunnels is estimated for different scenarios. The stochastic continuum approach has been investigated, as regards the representation of a scale dependent heterogeneous conductivity. An analytical method is proposed for the scaling of measured conductivity values, the method is consistent with the stochastic continuum approach. Some general conclusions from the work are: The larger the amount of heterogeneity, the larger the expected flow; The effects of the heterogeneity will decrease with increased tunnel length; If the conductivity of the tunnel is smaller than a threshold value, the tunnel conductivity is the most important parameter; If the tunnel conductivity is large and the tunnel is long, the most important parameter is the direction of the regional flow; Given a heterogeneous rock mass, if the tunnel length is shorter than about 500 m, the heterogeneity will be an important parameter, for lengths shorter than about 250 m, probably the most important; The flow through an encapsulation surrounded by a flow barrier is mainly dependent on the conductivity of the barrier. 70 refs, 110 figs, 10 tabs.

  20. Study of the distributions of flow rate and enthalpy in the sub-channels of a bundle geometry of nuclear reactors in one and two-phase flow

    International Nuclear Information System (INIS)

    Bayoumi, M.A.A.

    1976-10-01

    A bibliographic study shows that the experimental studies examined, have been developed to understand the phenomenon acting on the mixing between the sub-channels of which geometries are such these of rod bundles used in some nuclear reactors. Experimental devices and tests have been developed to study the influence of the following parameters, operating conditions, pressure, flow rate, power brought to the bundle and inlet temperature on the distribution of flow rates and vapor content among the different sub-channels. By means of non isokinetic sampling, one has determined the enthalpy of the fluid participating to the mixing between the communicating sub-channels and it has been shown that the value of this enthalpy depends strongly on the type of fluid flow and that this enthalpy cannot be either the enthalpy of one of the two sub-channels, nor (always) an average of these two enthalpies. The experimental results have been compared with calculations developed with the code FLICA, concerning the mass velocity distribution, the exchange term of linear momentum, and the variation of the transversal enthalpy with regard to the type of fluid flow. A study of local void ratio measurement, by means of optical probes, has been proposed. The present study has been carried out with a smooth geometry [fr

  1. Flow accelerated corrosion and its control measures for the secondary circuit pipelines in Indian nuclear power plants

    International Nuclear Information System (INIS)

    Kain, Vivekanand; Roychowdhury, S.; Mathew, Thomas; Bhandakkar, Atul

    2008-01-01

    A plain carbon steel feeder pipeline in the secondary circuit failed downstream of a flow measurement device (orifice meter) during operation at nuclear power plant. A detailed failure analysis done on the failed pipeline is described in this paper. The results established the fine surface pattern of 'Horseshoe pits' at the affected regions. X-ray diffraction analysis on the samples far from the failed regions showed presence of magnetite but on the sample from the failed region showed peaks due to base metal only, indicating dissolution of the oxide. Thickness profiling of the pipeline indicated reduction of thickness from the design 7.62 mm to a minimum of 0.4-1.4 mm at the location of the failure. These observations are characteristic of single phase flow accelerated corrosion. This paper details the extent of flow accelerated corrosion in various Indian power plants and the remedial measures for replacement and possible design and water chemistry changes to combat it

  2. Identification of two-phase flow patterns in a nuclear reactor by the high-frequency contribution fraction

    International Nuclear Information System (INIS)

    Wang, Y.W.; Pei, B.S.; King, C.H.; Lee, S.C.

    1989-01-01

    Recently, King et al. and Wang et al. analyzed the fluctuating characteristics of differential pressure and void fraction by the optimum modeling method and by spectral analysis, respectively. These two investigations presented some new concepts and deterministic criteria, which are based on purely empirical formulas, to identify two-phase flow patterns. These deterministic criteria on two-phase flow patterns' identification seem to show reasonable performance. In King's and Wang's studies, there are at least three problems that need further investigations for the applications to the nuclear reactor engineering field. These three problems are the following: 1. Is the response to a certain two-phase flow pattern, i.e., the fluctuating characteristics, of neutrons the same as that of differential pressure or void fraction? 2. Could those criteria developed from air/water flow be allowed to identify steam/water two-phase flow patterns? 3. Could those criteria be applied to identify two-phase flow patterns in rod bundles? In this paper, parts of the investigated results answer the first problem, and detailed comparisons with the previous work of the authors are given on a variety of items

  3. TYBO/BENHAM: Model Analysis of Groundwater Flow and Radionuclide Migration from Underground Nuclear Tests in Southwestern Pahute Mesa, Nevada

    Energy Technology Data Exchange (ETDEWEB)

    Andrew Wolfsberg; Lee Glascoe; Guoping Lu; Alyssa Olson; Peter Lichtner; Maureen McGraw; Terry Cherry; Guy Roemer

    2002-09-01

    Recent field studies have led to the discovery of trace quantities of plutonium originating from the BENHAM underground nuclear test in two groundwater observation wells on Pahute Mesa at the Nevada Test Site. These observation wells are located 1.3 km from the BENHAM underground nuclear test and approximately 300 m from the TYBO underground nuclear test. In addition to plutonium, several other conservative (e.g. tritium) and reactive (e.g. cesium) radionuclides were found in both observation wells. The highest radionuclide concentrations were found in a well sampling a welded tuff aquifer more than 500m above the BENHAM emplacement depth. These measurements have prompted additional investigations to ascertain the mechanisms, processes, and conditions affecting subsurface radionuclide transport in Pahute Mesa groundwater. This report describes an integrated modeling approach used to simulate groundwater flow, radionuclide source release, and radionuclide transport near the BENHAM and TYBO underground nuclear tests on Pahute Mesa. The components of the model include a flow model at a scale large enough to encompass many wells for calibration, a source-term model capable of predicting radionuclide releases to aquifers following complex processes associated with nonisothermal flow and glass dissolution, and site-scale transport models that consider migration of solutes and colloids in fractured volcanic rock. Although multiple modeling components contribute to the methodology presented in this report, they are coupled and yield results consistent with laboratory and field observations. Additionally, sensitivity analyses are conducted to provide insight into the relative importance of uncertainty ranges in the transport parameters.

  4. Cytometric analysis of shape and DNA content in mammalian sperm

    Energy Technology Data Exchange (ETDEWEB)

    Gledhill, B.L.

    1983-10-10

    Male germ cells respond dramatically to a variety of insults and are important reproductive dosimeters. Semen analyses are very useful in studies on the effects of drugs, chemicals, and environmental hazards on testicular function, male fertility and heritable germinal mutations. Sperm were analyzed by flow cytometry and slit-scan flow analysis for injury following the exposure of testes to mutagens. The utility of flow cytometry in genotoxin screening and monitoring of occupational exposure was evaluated. The technique proved valuable in separation of X- and Y-chromosome bearing sperm and the potential applicability of this technique in artificial insemination and a solution, of accurately assessing the DNA content of sperm were evaluated-with reference to determination of X- and Y-chromosome bearing sperm.

  5. Cytometric analysis of shape and DNA content in mammalian sperm

    International Nuclear Information System (INIS)

    Gledhill, B.L.

    1983-01-01

    Male germ cells respond dramatically to a variety of insults and are important reproductive dosimeters. Semen analyses are very useful in studies on the effects of drugs, chemicals, and environmental hazards on testicular function, male fertility and heritable germinal mutations. Sperm were analyzed by flow cytometry and slit-scan flow analysis for injury following the exposure of testes to mutagens. The utility of flow cytometry in genotoxin screening and monitoring of occupational exposure was evaluated. The technique proved valuable in separation of X- and Y-chromosome bearing sperm and the potential applicability of this technique in artificial insemination and a solution, of accurately assessing the DNA content of sperm were evaluated-with reference to determination of X- and Y-chromosome bearing sperm

  6. Define and Quantify the Physics of Air Flow, Pressure Drop and Aerosol Collection in Nuclear Grade HEPA Filters

    Energy Technology Data Exchange (ETDEWEB)

    Moore, Murray E. [Los Alamos National Lab. (LANL), Los Alamos, NM (United States)

    2015-02-23

    Objective: Develop a set of peer-review and verified analytical methods to adjust HEPA filter performance to different flow rates, temperatures and altitudes. Experimental testing will measure HEPA filter flow rate, pressure drop and efficiency to verify the analytical approach. Nuclear facilities utilize HEPA (High Efficiency Particulate Air) filters to purify air flow for workspace ventilation. However, the ASME AG-1 technical standard (Code on Nuclear Air and Gas Treatment) does not adequately describe air flow measurement units for HEPA filter systems. Specifically, the AG-1 standard does not differentiate between volumetric air flow in ACFM (actual cubic feet per minute)compared to mass flow measured in SCFM (standard cubic feet per minute). More importantly, the AG-1 standard has an overall deficiency for using HEPA filter devices at different air flow rates, temperatures, and altitudes. Technical Approach: The collection efficiency and pressure drops of 18 different HEPA filters will be measured over a range of flow rates, temperatures and altitudes. The experimental results will be compared to analytical scoping calculations. Three manufacturers have allocated six HEPA filters each for this effort. The 18 filters will be tested at two different flow rates, two different temperatures and two different altitudes. The 36 total tests will be conducted at two different facilities: the ATI Test facilities (Baltimore MD) and the Los Alamos National Laboratory (Los Alamos NM). The Radiation Protection RP-SVS group at Los Alamos has an aerosol wind tunnel that was originally designed to evaluate small air samplers. In 2010, modifications were started to convert the wind tunnel for HEPA filter testing. (Extensive changes were necessary for the required aerosol generators, HEPA test fixtures, temperature control devices and measurement capabilities.) To this date, none of these modification activities have been funded through a specific DOE or NNSA program. This is

  7. Validation and Calibration of Nuclear Thermal Hydraulics Multiscale Multiphysics Models - Subcooled Flow Boiling Study

    Energy Technology Data Exchange (ETDEWEB)

    Anh Bui; Nam Dinh; Brian Williams

    2013-09-01

    In addition to validation data plan, development of advanced techniques for calibration and validation of complex multiscale, multiphysics nuclear reactor simulation codes are a main objective of the CASL VUQ plan. Advanced modeling of LWR systems normally involves a range of physico-chemical models describing multiple interacting phenomena, such as thermal hydraulics, reactor physics, coolant chemistry, etc., which occur over a wide range of spatial and temporal scales. To a large extent, the accuracy of (and uncertainty in) overall model predictions is determined by the correctness of various sub-models, which are not conservation-laws based, but empirically derived from measurement data. Such sub-models normally require extensive calibration before the models can be applied to analysis of real reactor problems. This work demonstrates a case study of calibration of a common model of subcooled flow boiling, which is an important multiscale, multiphysics phenomenon in LWR thermal hydraulics. The calibration process is based on a new strategy of model-data integration, in which, all sub-models are simultaneously analyzed and calibrated using multiple sets of data of different types. Specifically, both data on large-scale distributions of void fraction and fluid temperature and data on small-scale physics of wall evaporation were simultaneously used in this work’s calibration. In a departure from traditional (or common-sense) practice of tuning/calibrating complex models, a modern calibration technique based on statistical modeling and Bayesian inference was employed, which allowed simultaneous calibration of multiple sub-models (and related parameters) using different datasets. Quality of data (relevancy, scalability, and uncertainty) could be taken into consideration in the calibration process. This work presents a step forward in the development and realization of the “CIPS Validation Data Plan” at the Consortium for Advanced Simulation of LWRs to enable

  8. Simplified flow cytometric assay to detect minimal residual disease in childhood with acute lymphoblastic leukemia Detecção de doença residual mínima em crianças com leucemia linfoblástica aguda por citometria de fluxo

    Directory of Open Access Journals (Sweden)

    Elizabete Delbuono

    2008-08-01

    Full Text Available The detection of minimal residual disease (MRD is an important prognostic factor in childhood acute lymphoblastic leukemia (ALL providing crucial information on the response to treatment and risk of relapse. However, the high cost of these techniques restricts their use in countries with limited resources. Thus, we prospectively studied the use of flow cytometry (FC with a simplified 3-color assay and a limited antibody panel to detect MRD in the bone marrow (BM and peripheral blood (PB of children with ALL. BM and PB samples from 40 children with ALL were analyzed on days (d 14 and 28 during induction and in weeks 24-30 of maintenance therapy. Detectable MRD was defined as > 0.01% cells expressing the aberrant immunophenotype as characterized at diagnosis among total events in the sample. A total of 87% of the patients had an aberrant immunophenotype at diagnosis. On d14, 56% of the BM and 43% of the PB samples had detectable MRD. On d28, this decreased to 45% and 31%, respectively. The percentage of cells with the aberrant phenotype was similar in both BM and PB in T-ALL but about 10 times higher in the BM of patients with B-cell-precursor ALL. Moreover, MRD was detected in the BM of patients in complete morphological remission (44% on d14 and 39% on d28. MRD was not significantly associated to gender, age, initial white blood cell count or cell lineage. This FC assay is feasible, affordable and readily applicable to detect MRD in centers with limited resources.A detecção de doença residual mínima (DRM é um importante fator prognóstico na leucemia linfóide aguda (LLA infantil e fornece informações sobre a resposta ao tratamento e o risco de recaída. Entretanto, os altos custos das técnicas utilizadas limitam seu uso nos países em desenvolvimento. Desta forma, realizamos um estudo prospectivo para avaliar a citometria de fluxo (CF, utilizando três fluorescências e um painel limitado de anticorpos monoclonais, como método de detec

  9. Development of a model and test equipment for cold flow tests at 500 atm of small nuclear light bulb configurations

    Science.gov (United States)

    Jaminet, J. F.

    1972-01-01

    A model and test equipment were developed and cold-flow-tested at greater than 500 atm in preparation for future high-pressure rf plasma experiments and in-reactor tests with small nuclear light bulb configurations. With minor exceptions, the model chamber is similar in design and dimensions to a proposed in-reactor geometry for tests with fissioning uranium plasmas in the nuclear furnace. The model and the equipment were designed for use with the UARL 1.2-MW rf induction heater in tests with rf plasmas at pressures up to 500 atm. A series of cold-flow tests of the model was then conducted at pressures up to about 510 atm. At 504 atm, the flow rates of argon and cooling water were 3.35 liter/sec (STP) and 26 gal/min, respectively. It was demonstrated that the model is capable of being operated for extended periods at the 500-atm pressure level and is, therefore, ready for use in initial high-pressure rf plasma experiments.

  10. An integrated model for reliability estimation of digital nuclear protection system based on fault tree and software control flow methodologies

    International Nuclear Information System (INIS)

    Kim, Man Cheol; Seong, Poong Hyun

    2000-01-01

    In the nuclear industry, the difficulty of proving the reliabilities of digital systems prohibits the widespread use of digital systems in various nuclear application such as plant protection system. Even though there exist a few models which are used to estimate the reliabilities of digital systems, we develop a new integrated model which is more realistic than the existing models. We divide the process of estimating the reliability of a digital system into two phases, a high-level phase and a low-level phase, and the boundary of two phases is the reliabilities of subsystems. We apply software control flow method to the low-level phase and fault tree analysis to the high-level phase. The application of the model to Dynamic Safety System(DDS) shows that the estimated reliability of the system is quite reasonable and realistic

  11. Modeling three dimensional flows in the lower plenum of Loviisa nuclear power plant with the CFX code

    International Nuclear Information System (INIS)

    Bernard, J.P.; Haekkinen, J.; Sarkomaa, P.

    1997-01-01

    A number of numerical studies on three-dimensional flows in reactor vessels of nuclear power plants have been carried out recently. The main reason for this is the phenomenon of inherent boron dilution. These studies have been done for Loviisa nuclear power plant as well. This presentation focuses on the three dimensional modeling of Loviisa's lower plenum with the commercial CFX code. The investigations particularly deal with the modeling of the two perforated plates located at the bottom and just below the core of the vessel. The perforated plates can be modeled as porous media in the CFX code. The model has been validated against available experimental data. Velocity profiles around the plates, the pressure drop through the plates, and the simulation of mixing factors have been investigated specifically. (author)

  12. Computations of steam flow and heat transfer in nuclear power plant condensers

    International Nuclear Information System (INIS)

    Yuan, A.

    1997-01-01

    To improve performance of its PWR nuclear power plants, Electricite de France has developed a performance monitoring system that checks simultaneously the operation of the components of the secondary system. The performance monitoring system is based on a computational software CITER for steady state runs. A one-dimensional condenser model has been developed. Application of this code to a nuclear power plant condenser shows that predicted values in good agreement with the design values

  13. Development and computational simulation of thermoelectric electromagnetic pumps for controlling the fluid flow in liquid metal cooled space nuclear reactors

    International Nuclear Information System (INIS)

    Borges, E.M.

    1991-01-01

    Thermoelectric Electromagnetic (TEEM) Pumps can be used for controlling the fluid flow in the primary and secondary circuits of liquid metal cooled space nuclear reactor. In order to simulate and to evaluate the pumps performance, in steady-state, the computer program BEMTE has been developed to study the main operational parameters and to determine the system actuation point, for a given reactor operating power. The results for each stage of the program were satisfactory, compared to experimental data. The program shows to be adequate for the design and simulating of direct current electromagnetic pumps. (author)

  14. Influence of the nuclear symmetry energy on the collective flows of charged pions

    Science.gov (United States)

    Gao, Yuan; Yong, Gao-Chan; Zhang, Lei; Zuo, Wei

    2018-01-01

    Based on the isospin-dependent Boltzmann-Uehling-Uhlenbeck (IBUU) transport model, we studied charged pion transverse and elliptic flows in semicentral 197Au+197Au collisions at 600 MeV/nucleon. It is found that π+-π- differential transverse flow and the difference of π+ and π- transverse flows almost show no effects of the symmetry energy. Their corresponding elliptic flows are largely affected by the symmetry energy, especially at high transverse momenta. The isospin-dependent pion elliptic flow at high transverse momenta thus provides a promising way to probe the high-density behavior of the symmetry energy in heavy-ion collisions at the Facility for Antiproton and Ion Research (FAIR) at GSI, Darmstadt or at the Cooling Storage Ring (CSR) at HIRFL, Lanzhou.

  15. Flow

    DEFF Research Database (Denmark)

    Knoop, Hans Henrik

    2006-01-01

    FLOW. Orden i hovedet på den fede måde Oplevelsesmæssigt er flow-tilstanden kendetegnet ved at man er fuldstændig involveret, fokuseret og koncentreret; at man oplever stor indre klarhed ved at vide hvad der skal gøres, og i hvilket omfang det lykkes; at man ved at det er muligt at løse opgaven...

  16. Numerical simulation on flow field of nuclear safety grade 2 single-seat pneumatic diaphragm control valve

    International Nuclear Information System (INIS)

    Zhong Yun; Zhang Jige; Wang Dezhong; Shi Jianzhong

    2010-01-01

    The Computational Fluid Dynamics (CFD) method is employed to simulate numerically the steady flow and transient flow under variable openings of the nuclear safety grade 2 single-seat pneumatic diaphragm control valve, which is a sleeve valve. The steady simulations under rated condition tells that there is a large amount of vortex in the valve seat necking and around the valve cone, which leads to a much greater flow impact on the head of the valve cone and uneven pressure distribution on spool face. More consideration should be taken on the characteristics of the valve cone accordingly, when designing a valve of this kind. Then the transient flow under 100% and 40% openings is simulated numerically on the basis of steady simulations. The pulsation of the pressure magnitude at the points with large vorticity, in the valve seat necking and around the valve cone, is monitored. The main pulsation frequencies differ from the low natural frequencies of the model, which means that it is safe from leading to structural resonance. (authors)

  17. Flow modeling in heterogeneous media in the context of geologic nuclear waste repositories

    International Nuclear Information System (INIS)

    Sagar, B.

    1995-01-01

    Assessment of long term-performance of geologic repositories requires simulation of flow through heterogeneous geologic formations. Effect on flow field of discontinuities such as fracture zones, in such media is of interest to not only waste management but also in petroleum engineering and water resources development. In this paper, a technique of deriving mass balance equations in the presence of fractures is discussed. Compared to full representation of fractures, the proposed technique provides coarser resolution of the flow field but it is relatively computationally efficient. Two examples of its application are also provided

  18. Supercontinuum white light lasers for flow cytometry

    OpenAIRE

    Telford, William G.; Subach, Fedor V.; Verkhusha, Vladislav V.

    2009-01-01

    Excitation of fluorescent probes for flow cytometry has traditionally been limited to a few discrete laser lines, an inherent limitation in our ability to excite the vast array of fluorescent probes available for cellular analysis. In this report, we have used a supercontinuum (SC) white light laser as an excitation source for flow cytometry. By selectively filtering the wavelength of interest, almost any laser wavelength in the visible spectrum can be separated and used for flow cytometric a...

  19. Long-term storage of samples for flow cytometric DNA analysis

    DEFF Research Database (Denmark)

    Vindeløv, L L; Christensen, I J; Keiding, N

    1983-01-01

    estimation by deconvolution, there was significant intraday and interday variation. Hence the most accurate results are obtained if different aliquots of a sample are measured on different days rather than on the same day. Use of the storage method thus has the potential of increasing the accuracy...

  20. Flow-cytometric measurements of somatic cell mutations in Thorotrast patients

    International Nuclear Information System (INIS)

    Umeki, Shigeko; Kyoizumi, Seishi; Kusunoki, Yoichiro; Nakamura, Nori; Sasaki, Masao; Mori, Takesaburo; Ishikawa, Yuichi; Cologne, J.B.; Akiyama, Mitoshi.

    1992-10-01

    Exposure to ionizing radiation is a well-recognized risk factor for cancer development. Because ionizing radiation can induce mutations, an accurate way of measuring somatic mutation frequencies could be a useful tool for evaluating cancer risk. In the present study, we have examined in vivo somatic mutation frequencies at the erythrocyte glycophorin A and T-cell receptor loci in 18 Thorotrast patients. These persons have been continuously irradiated with alpha particles emitted from the internal deposition of thorium dioxide and thus have increased risks of certain malignant tumors. When compared with controls, the Thorotrast patients showed a significantly higher frequency of mutants at the lymphocyte T-cell receptor loci but not at the erythrocyte glycophorin A loci. (author)

  1. DEA 1 Expression on Dog Erythrocytes Analyzed by Immunochromatographic and Flow Cytometric Techniques

    OpenAIRE

    Acierno, M.M.; Raj, K.; Giger, U.

    2014-01-01

    Background The Dog erythrocyte antigen (DEA) 1 blood group system was thought to contain types DEA 1.1 and 1.2 (and possibly 1.3 [A3]). However, DEA 1.2+ dogs are very rare and newer typing methods reveal varying degrees of DEA 1 positivity. Objectives To assess if variation in DEA 1 positivity is because of quantitative differences in surface antigen expression. To determine expression patterns in dogs over time and effects of blood storage (4?C). To evaluate DEA 1.2+ samples by DEA 1 typing...

  2. Flow cytometric analysis of lymphocytes in aplastic anemia among atomic bomb survivors

    International Nuclear Information System (INIS)

    Imamura, Nobutaka; Inada, Tominari; Asaoku, Hideki; Abe, Kazuhiro; Oguma, Nobuo; Kuramoto, Atsushi

    1986-01-01

    In 6 patients with aplastic anemia and 3 patients with pernicious anemia, lymphocyte subpopulations in the peripheral blood were measured, before and after steroid therapy, with a fluorescence-activated cell sorder using various monoclonal antibodies. The ratio of OKT4-positive lymphocytes (T4) to OKT8-positive lymphocytes (T8) in the peripheral blood was reduced in 2 patients (20 %). The T4/T8 ratio returned to normal during remission of anemia. Hematological improvement was seen after a large amount of steroid therapy in 3 patients. The number of Tac-positive cells tended to decrease and the T4/T8 ratio tended to return to normal with hematological improvement, although there was no correlation to hydrocortisone reaction. Some patients were supposed to have abnormal number of suppressor and inducer T cells. (Namekawa, K.)

  3. Flow cytometric analysis of RNA synthesis by detection of bromouridine incorporation

    DEFF Research Database (Denmark)

    Larsen, J K; Jensen, Peter Østrup; Larsen, J

    2001-01-01

    RNA synthesis has traditionally been investigated by a laborious and time-consuming radiographic method involving incorporation of tritiated uridine. Now a faster non-radioactive alternative has emerged, based on immunocytochemical detection. This method utilizes the brominated RNA precursor...... bromouridine, which is taken into a cell, phosphorylated, and incorporated into nascent RNA. The BrU-substituted RNA is detected by permeabilizing the cells and staining with certain anti-BrdU antibodies. This dynamic approach yields information complementing that provided by cellular RNA content analysis...

  4. Flow cytometric assay detecting cytotoxicity against human endogenous retrovirus antigens expressed on cultured multiple sclerosis cells

    DEFF Research Database (Denmark)

    Møller-Larsen, A; Brudek, T; Petersen, T

    2013-01-01

    expressing increased amounts of human endogenous retrovirus antigens. MS patients also have increased antibody levels to these antigens. The target cells are spontaneously growing peripheral blood mononuclear cells (PBMCs) of B cell lineage, expressing human endogenous retrovirus HERV epitopes...... on their surface. Polyclonal antibodies against defined peptides in the Env- and Gag-regions of the HERVs were raised in rabbits and used in antibody-dependent cell-mediated cytotoxicity (ADCC) -assays. Rituximab® (Roche), a chimeric monoclonal antibody against CD20 expressed primarily on B cells, was used...

  5. Proliferation and apoptosis in infection with infectious bursal disease virus: a flow cytometric study.

    Science.gov (United States)

    Ojeda, F; Skardova, I; Guarda, M I; Ulloa, J; Folch, H

    1997-01-01

    Programmed cell death, or apoptosis, is involved in the normal physiology of many immunocompetent organs, including lymphocytes of the bursa of Fabricius in chickens. Involvement of apoptosis has also been described in some viral diseases such as AIDS. The purpose of this work was to study the potential role of apoptosis in the pathogenesis of Gumboro disease in the bursa of Fabricius. Our results show that 1-3 days after infection of young chickens with infectious bursal disease virus, the number of apoptotic cells increases and cellularity and proliferation decrease. Because of the dynamic nature of bursal lymphocyte populations and the involvement of apoptosis in lymphocyte cell physiology, the increased level of cells undergoing apoptosis may be due to an impairment in the withdrawal of apoptotic cells. A concomitant increase in macrophages in infected bursae and a dramatic decrease in cellularity suggest that an increase in apoptosis may be an important cause of cell depletion.

  6. On the use of nuclear magnetic resonance to characterize vertical two-phase bubbly flows

    International Nuclear Information System (INIS)

    Lemonnier, H.; Jullien, P.

    2011-01-01

    Research highlights: → We provide a complete theory of the PGSE measurement in single and two-phase flow. → Friction velocity can be directly determinated from measured velocity distributions. → Fast determination of moments shorten PGSE process with small loss of accuracy. → Turbulent diffusion measurements agree well with known trends and existing models. → We think NMR can be a tool to benchmark thermal anemometry in two-phase flow. - Abstract: Since the pioneering work of who showed that NMR can be used to measure accurately the mean liquid velocity and void fraction in two-phase pipe flow, it has been shown that NMR signal can also characterize the turbulent eddy diffusivity and velocity fluctuations. In this paper we provide an in depth validation of these statements together with a clarification of the nature of the mean velocity that is actually measured by NMR PFGSE sequence. The analysis shows that the velocity gradient at the wall is finely space-resolved and allows the determination of the friction velocity in single-phase flows. Next turbulent diffusion measurements in two-phase flows are presented, analyzed and compared to existing data and models. It is believed that NMR velocity measurement is sufficiently understood that it can be utilized to benchmark thermal anemometry in two-phase flows. Theoretical results presented in this paper also show how this can be undertaken.

  7. Optimization of material flow in the nuclear fuel cycle using a cyclic multi-stage production-to-inventory model

    International Nuclear Information System (INIS)

    DePorter, E.L.

    1977-01-01

    The nuclear fuel cycle is modelled as a cyclic, multi-stage production-to-inventory system. The objective is to meet a known deterministic demand for energy while minimizing acquisition, production, and inventory holding costs for all stages of the fuel cycle. The model allows for cyclic flow (feedback) of materials, material flow conversion factors at each stage, production lag times at each stage, and for escalating costs of uranium ore. It does not allow shortages to occur in inventories. The model is optimized by the application of the calculus of variations and specifically through recently developed theorems on the solution of functionals constrained by inequalities. The solution is a set of optimal cumulative production trajectories which define the stagewise production rates. Analysis of these production rates reveals the optimal nuclear fuel cycle costs and that inventories (stockpiles) occur in uranium fields, enriched uranium hexafluoride, and fabricated fuel assemblies. An analysis of the sensitivity of the model to variation in three important parameters is performed

  8. Determination of Ploidy Level and Nuclear DNA Content in the Droseraceae by Flow Cytometry

    Czech Academy of Sciences Publication Activity Database

    Hoshi, Y.; Azumatani, M.; Suyama, T.; Adamec, Lubomír

    2017-01-01

    Roč. 82, č. 3 (2017), s. 321-327 ISSN 0011-4545 Institutional support: RVO:67985939 Keywords : nuclear DNA content * genome size * Droseraceae Subject RIV: EB - Genetics ; Molecular Biology OBOR OECD: Plant sciences, botany Impact factor: 0.913, year: 2016

  9. Differential gene flow of mitochondrial and nuclear DNA markers among chromosomal races of Australian morabine grasshoppers (Vandiemenella, viatica species group).

    Science.gov (United States)

    Kawakami, T; Butlin, R K; Adams, M; Saint, K M; Paull, D J; Cooper, S J B

    2007-12-01

    Recent theoretical developments have led to a renewed interest in the potential role of chromosomal rearrangements in speciation. Australian morabine grasshoppers (genus Vandiemenella, viatica species group) provide an excellent study system to test this potential role of chromosomal rearrangements because they show extensive chromosomal variation and formed the basis of a classic chromosomal speciation model. There are three chromosomal races, viatica19, viatica17, and P24(XY), on Kangaroo Island, South Australia, forming five parapatric populations with four putative contact zones among them. We investigate the extent to which chromosomal variation among these populations may be associated with barriers to gene flow. Population genetic and phylogeographical analyses using 15 variable allozyme loci and the elongation factor-1alpha (EF-1alpha) gene indicate that the three races represent genetically distinct taxa. In contrast, analyses of the mitochondrial cytochrome c oxidase subunit I (COI) gene show the presence of three distinctive and geographically localized groups that do not correspond with the distribution of the chromosomal races. These discordant population genetic patterns are likely to result from introgressive hybridization between the chromosomal races and range expansions/contractions. Overall, these results suggest that reduction of nuclear gene flow may be associated with chromosomal variation, or underlying genetic variation linked with chromosomal variation, whereas mitochondrial gene flow appears to be independent of this variation in these morabine grasshoppers. The identification of an intact contact zone between P24(XY) and viatica17 offers considerable potential for further investigation of molecular mechanisms that maintain distinct nuclear genomes among the chromosomal races.

  10. Cytometric analysis of DNA changes induced by sulfur mustard

    Energy Technology Data Exchange (ETDEWEB)

    Smith, W.J.; Sanders, K.M.; Ruddle, S.E.; Gross, C.L.

    1993-05-13

    Sulfur mustard is an alkylating agent which causes severe, potentially debilitating blisters following cutaneous exposure. Its mechanism of pathogenesis is unknown and no antidote exists to prevent its pathology. The biochemical basis of sulfur mustard's vesicating activity has been hypothesized to be a cascade of events beginning with alkylation of DNA. Using human cells in culture, we have assessed the effects of sulfur mustard on cell cycle activity using flow cytometry with propidium iodide. Two distinct patterns emerged, a Gl/S interface block at concentrations equivalent to vesicating doses (>50-micronM) and a G2 block at 10-fold lower concentrations. In addition, noticeable increases in amount of dye uptake were observed at 4 and 24 hours after sulfur mustard exposure. These increases are believed to be related to DNA repair activities and can be prevented by treatment of the cells with niacinamide, which inhibits DNA repair. Other drugs which provide alternate alkylating sites or inhibit cell cycle progression were shown to lower the cytotoxicity of sulfur mustard and to protect against its direct DNA damaging effects.

  11. Parameters influencing flow-accelerated corrosion (FAC) in the secondary side of nuclear power plants

    International Nuclear Information System (INIS)

    Khatibi, M.; Lister, D.H.; Feicht, A.; Liu, L.; Uchida, S.; Fujiwara, K.; Ohira, T.; Takiguchi, H.; Hisanume, K.

    2009-01-01

    Thermal power plants have reported excessive pipe degradation because of Flow-Accelerated Corrosion (FAC) since the 1960s. Common features have been the use of carbon steel in regions of high flow rate and high turbulence with a water chemistry of modest alkalinity and free of oxidizing agents. It can be concluded that the main parameters that affect FAC are flow dynamics, water chemistry and composition of the materials used in pipework and components. A clear indication of FAC is the rapid wall thinning, usually in the presence of distinct flow-related markings, or scallops, on the surface. On more than one occasion, FAC has been responsible for large pipe failures that have led to serious damage and in some cases fatalities. After such a failure at the Mihama-3 PWR in 2004, a collaborative research program between Canada and Japan was initiated to improve the understanding of FAC by studying both the individual and the synergistic effects of feed-water system parameters. In an experimental water loop, three test sections were installed in series. Test section 1 contained probes made of the carbon steel of interest to measure on-line the FAC rate and electrochemical corrosion potential (ECP). Test sections 2 and 3 contained surface analysis probes for examination after removal via optical, SEM and Raman, techniques. The effects of flow and other parameters on FAC were studied using probes of different bore size, different material and several flow rates. Experiments were performed under neutral and ammoniated chemistries in de-oxygenated and oxygenated water. Threshold concentrations of oxygen to stifle FAC were determined. The individual and combined effects of system variables have now been determined in some detail and are presented here. (author)

  12. RECENT IMPROVEMENTS IN THE CUPID CODE FOR A MULTI-DIMENSIONAL TWO-PHASE FLOW ANALYSIS OF NUCLEAR REACTOR COMPONENTS

    Directory of Open Access Journals (Sweden)

    HAN YOUNG YOON

    2014-10-01

    Full Text Available The CUPID code has been developed at KAERI for a transient, three-dimensional analysis of a two-phase flow in light water nuclear reactor components. It can provide both a component-scale and a CFD-scale simulation by using a porous media or an open media model for a two-phase flow. In this paper, recent advances in the CUPID code are presented in three sections. First, the domain decomposition parallel method implemented in the CUPID code is described with the parallel efficiency test for multiple processors. Then, the coupling of CUPID-MARS via heat structure is introduced, where CUPID has been coupled with a system-scale thermal-hydraulics code, MARS, through the heat structure. The coupled code has been applied to a multi-scale thermal-hydraulic analysis of a pool mixing test. Finally, CUPID-SG is developed for analyzing two-phase flows in PWR steam generators. Physical models and validation results of CUPID-SG are discussed.

  13. Wind tunnel experimental study on effect of inland nuclear power plant cooling tower on air flow and dispersion of pollutant

    International Nuclear Information System (INIS)

    Qiao Qingdang; Yao Rentai; Guo Zhanjie; Wang Ruiying; Fan Dan; Guo Dongping; Hou Xiaofei; Wen Yunchao

    2011-01-01

    A wind tunnel experiment for the effect of the cooling tower at Taohuajiang nuclear power plant on air flow and dispersion of pollutant was introduced in paper. Measurements of air mean flow and turbulence structure in different directions of cooling tower and other buildings were made by using an X-array hot wire probe. The effects of the cooling tower and its drift on dispersion of pollutant from the stack were investigated through tracer experiments. The results show that the effect of cooling tower on flow and dispersion obviously depends on the relative position of stack to cooling towers, especially significant for the cooling tower parallel to stack along wind direction. The variation law of normalized maximum velocity deficit and perturbations in longitudinal turbulent intensity in cooling tower wake was highly in accordance with the result of isolated mountain measured by Arya and Gadiyaram. Dispersion of pollutant in near field is significantly enhanced and plume trajectory is changed due to the cooling towers and its drift. Meanwhile, the effect of cooling tower on dispersion of pollutant depends on the height of release. (authors)

  14. Measurement and modeling of flow through unsaturated heterogeneous rock in the context of geologic disposal of nuclear waste

    International Nuclear Information System (INIS)

    Sagar, B.; Bagtzoglou, A.C.; Green, R.T.; Stothoff, S.A.

    1995-01-01

    Deep geologic disposal of high-level and transuranic waste is currently being pursued vigorously. Assessing long-term performance of such repositories involves laboratory and field measurements, and numerical modeling. There exist two primary characteristics, associated with assessing repository performance, that define problems of modeling and measurement of non-isothermal flow through geologic media exposed to variable boundary conditions (e.g., climatic changes). These are: (1) the large time scale (tens of thousands of years) and highly variable space scale (from one meter to 10 5 meters); and (2) the hierarchy of heterogeneities and discontinuities characterizing the medium. This paper provides an overview of recent work, conducted at the Center for Nuclear Waste Regulatory Analyses (CNWRA), related to laboratory experiments, consideration of similitude, and numerical modeling of flow through heterogeneous media under non-homogeneous boundary conditions. As discussed, there exist neither good methods of measuring flows at these scales nor are there adequate similitude analyses that would allow reasonable scaling up of laboratory-scale experiments. Reliable assessment of long-term geologic repositories will require sophisticated geostatistical models capable of addressing variables scales of heterogeneities conditioned with observed results from adequately sized field-scale experiments conducted for sufficiently long durations

  15. Flow instability tests for a particle bed reactor nuclear thermal rocket fuel element

    Science.gov (United States)

    Lawrence, Timothy J.

    1993-05-01

    Recent analyses have focused on the flow stability characteristics of a particle bed reactor (PBR). These laminar flow instabilities may exist in reactors with parallel paths and are caused by the heating of the gas at low Reynolds numbers. This phenomena can be described as follows: several parallel channels are connected at the plenum regions and are stabilized by some inlet temperature and pressure; a perturbation in one channel causes the temperature to rise and increases the gas viscosity and reduces the gas density; the pressure drop is fixed by the plenum regions, therefore, the mass flow rate in the channel would decrease; the decrease in flow reduces the ability to remove the energy added and the temperature increases; and finally, this process could continue until the fuel element fails. Several analyses based on different methods have derived similar curves to show that these instabilities may exist at low Reynolds numbers and high phi's ((Tfinal Tinitial)/Tinitial). These analyses need to be experimentally verified.

  16. Spent Nuclear Fuel (SNF) Project Canister Storage Building (CSB) Process Flow Diagram Mass Balance Calculations

    International Nuclear Information System (INIS)

    KLEM, M.J.

    2000-01-01

    The purpose of these calculations is to develop the material balances for documentation of the Canister Storage Building (CSB) Process Flow Diagram (PFD) and future reference. The attached mass balances were prepared to support revision two of the PFD for the CSB. The calculations refer to diagram H-2-825869

  17. Spent Nuclear Fuel (SNF) Project Multi Canister Overpack (MCO) Process Flow Diagram Mass Balance Calculations

    International Nuclear Information System (INIS)

    KLEM, M.J.

    2000-01-01

    The purpose of this calculation document is to develop the bases for the material balances of the Multi-Canister Overpack (MCO) Level 1 Process Flow Diagram (PFD). The attached mass balances support revision two of the PFD for the MCO and provide future reference

  18. Equations of motion for two-phase flow in a pin bundle of a nuclear reactor

    International Nuclear Information System (INIS)

    Chawla, T.C.; Ishii, M.

    1978-01-01

    By performing Eulerian area averaging over a channel area of the local continuity, momentum, and energy equations for single phase turbulent flow and assuming each phase in two-phase flows to be continuum but coupled by the appropriate 'jump' conditions at the interface, the corresponding axial macroscopic balances for two-fluid model in a pin bundle are obtained. To determine the crossflow, a momentum equation in transverse (to the gap between the pins) direction is obtained for each phase by carrying out Eulerian segment averaging of the local momentum equation, where the segment is taken parallel to the gap. By considering the mixture as a whole, a diffusion model based on drift-flux velocity is formulated. In the axial direction it is expressed in terms of three mixture conservation equations of mass, momentum, and energy with one additional continuity equation for the vapor phase. For the determination of crossflow, transverse momentum equation for a mixture is obtained. It is considered that the previous formulation of the two-phase flow based on the 'slip' flow model and the integral subchannel balances using finite control volumes is inadequate in that the model is heuristic and, a priori, assumes the order of magnitude of the terms, also the model is incomplete and incorrect when applied to two-phase mixtures in thermal non-equilibrium such as during accidental depressurization of a water cooled reactor. The governing equations presented are shown to be a very formal and sound physical basis and are indispensable for physically correct methods of analyzing two-phase flows in a pin bundle. (author)

  19. The study of flow resistance in nuclear reactor Maria under coolant boiling condition

    International Nuclear Information System (INIS)

    Czerski, P.

    1999-01-01

    This study describes an analysis of experiments carried out in the WIW-300 installation located in the Institute of Atomic Energy (Swierk, Poland). The flow, simulated in the annular gap of test section, was similar to the flow in Maria reactor fuel channel. Experimental character of the work lead to the conclusions related to the physical nature of the hydrodynamic phenomena investigated as well as to the practical aspects of future research. A hypothesis defining a cause of pressure changes was formulated and specific problems related to the mathematical model were defined. The analysis shows that hydrodynamic phenomena studies are of basic significance for the prediction of burnout effects and that heat exchange is very often determined by local phenomena. All described observations are the base for further research on thermodynamic aspects of investigated phenomena. (author)

  20. PANTHERE, simulation software of dose flow rates for complex nuclear installations

    International Nuclear Information System (INIS)

    Longeot, M.; Dupont, B.; Coatanea, C.; Schumm, S.; Zweers, M.; Malvagi, F.; Trama, J.C.

    2010-01-01

    The authors present the PANTHERE simulation software developed by EDF-SEPTEN to determine gamma dose flow rate in any point of complex industrial installations. They present the current industrial version (PANTHEREV1) and its different applications, and more particularly an investigation in the field of qualification of hardware under irradiation in case of severe accident. They present the currently under development version (PANTHEREV2) which will be exploited in 2011

  1. Correlations between wind flow and population location at 67 light water nuclear power plant sites. [USA

    Energy Technology Data Exchange (ETDEWEB)

    Sprung, J.L.; Steck, G.P.; Frazier, A.W.

    1978-10-01

    Because wind flow and population location are both likely to be influenced by topography, it has been suggested that wind distributions and population distributions ought to be correlated and that the neglect of these correlations in the calculations of the Reactor Safety Study could have resulted in significant underestimates of accident consequences. This paper presents the results of an investigation of correlations between wind roses and population locations at 67 of the 68 power plant sites included in the Reactor Safety Study.

  2. Fluid flows due to leakproofness defects of nuclear fuel cans: application to sipping in PWR

    International Nuclear Information System (INIS)

    Parrat, D.

    1983-10-01

    This thesis establishes the theoritical laws describing fluid and gas transfers through channels representative of nuclear fuel can defects. These laws, applied to craks on fuel cans (samples) and on new or spent experimental rods, allow to know their size. A computerized modelling has been developed to analyse more particularly the examination by ''quantitative permeation testing'', giving the equivalent size of a channel from the observed signal. The utilization of this process to non-tight fuel assemblies of pressurized water reactors leads to the establishment of a classification of defects according to their importance [fr

  3. Prediction of volume fractions in three-phase flows using nuclear technique and artificial neural network

    International Nuclear Information System (INIS)

    Marques Salgado, Cesar; Brandao, Luis E.B.; Schirru, Roberto; Pereira, Claudio M.N.A.; Silva, Ademir Xavier da; Ramos, Robson

    2009-01-01

    This work presents methodology based on nuclear technique and artificial neural network for volume fraction predictions in annular, stratified and homogeneous oil-water-gas regimes. Using principles of gamma-ray absorption and scattering together with an appropriate geometry, comprised of three detectors and a dual-energy gamma-ray source, it was possible to obtain data, which could be adequately correlated to the volume fractions of each phase by means of neural network. The MCNP-X code was used in order to provide the training data for the network.

  4. The Experience at Russian Nuclear Sites of Modeling Groundwater Flow on Different Scales

    Science.gov (United States)

    Zinin, A.; Zinina, G.; Samsanova, L.; Vasilkova, N.; Alexandrova, L.; Drozhko, E.

    2001-12-01

    The experience of developing models of different scales to predict contaminant plume migration in ground waters is analyzed. The method of developing a three-dimensional transient model is demonstrated to estimate high-density solutions migrating from the surface storage of liquid radioactive waste, using a two-dimensional regional model for setting boundary conditions (Lake Karachay, PA "Mayak", Russia). The model is used to calculate three-dimensional transient distribution of pressure, density and concentrations of the dissolved admixtures in the non-confined aquifers. Interpolation is also specified to calculate boundary conditions parameters of the inserted models. The method of constructing a local filtration model is described to predict the contaminant plume spreading from the operating ground of deep burial of liquid radioactive wastes (The Siberian Chemical Plant, Seversk). The local model uses smaller grid gaps over time and space and a more detailed stratiographic division of the section as compared to the regional model intended to be used for estimating groundwater resourses. The flow distribution within the local model boundaries is described as the products of an average annual flow and periodical time function (function of monthly fluctuations) and the function of spatial variables. The parameters of the distribution function, represented on the local model grid by the values, were determined by solving the inverse problem. The sensivity analysis of the target function of the inverse problem to the small variations of the average annual flows is described.

  5. Subcadmic and epicadmic flow in the dry tube of the TRIGA Mark III reactor of the Nuclear Center of Mexico

    International Nuclear Information System (INIS)

    Delfin L, A.; Mazon R, R.; Nava R, B.

    1991-04-01

    The mensuration of the thermal and fast flows of the irradiation facilities of the core of the reactor is important, since allow us to determine the optimum time of irradiation of the samples in the reactor. The Dry Tube especially, is an irradiation installation that it was designed in the I.N.I.N. to supply the pneumatic irradiation system of capsules with durations bigger than 15 minutes and it can be used for exposures until a maximum of three hours. The main users are the Nuclear Chemistry Department and the Neutron activation analysis. In this report the neutron flux sub cadmic and epi cadmic obtained in an experimental way in the Dry Tube for the reactor operating in stationary state to powers of 100 Kw, 300 Kw and 1000 Kw are reported and with these values it is interpolated for other powers. (Author)

  6. Citizen perceptions of information flow around a nuclear facility: A study in risk communication

    International Nuclear Information System (INIS)

    Price, M.O.

    1997-01-01

    Responses of focus group members from the region around a Nuclear Facility provide the data for this qualitative study concerning citizen perceptions of available site information. Analyses of three of the focus group discussion questions and the answers they elicited showed a dominant perception among participants of insufficient easily available information about the site. These respondents also indicated that most of them obtain site information through mass media and hearsay, that many lack trust in the information they have and would trust only an independent entity to provide accurate information. A new area in communication studies, variously called environmental risk communication, risk communication and health risk communication, continues to evolve among those working in various allied disciplines, some far removed from communication. As science attempts to solve environmental problems caused by technological advances, this field acquires numerous practitioners. Some of these risk communication experts may however, be overlooking basic and necessary components of effective communication, because their expertise is in another discipline. One result of this can be communication breakdown in which those involved, assume that meaning is shared, when in fact the opposite is true. This paper seeks to clarify a necessary ingredient of effective interpersonal risk communication, using data obtained from citizens living around one of the nation's nuclear facilities as an example

  7. Flow Accelerated Erosion-Corrosion (FAC) considerations for secondary side piping in the AP1000R nuclear power plant design

    International Nuclear Information System (INIS)

    Vanderhoff, J. F.; Rao, G. V.; Stein, A.

    2012-01-01

    The issue of Flow Accelerated Erosion-Corrosion (FAC) in power plant piping is a known phenomenon that has resulted in material replacements and plant accidents in operating power plants. Therefore, it is important for FAC resistance to be considered in the design of new nuclear power plants. This paper describes the design considerations related to FAC that were used to develop a safe and robust AP1000 R plant secondary side piping design. The primary FAC influencing factors include: - Fluid Temperature - Pipe Geometry/layout - Fluid Chemistry - Fluid Velocity - Pipe Material Composition - Moisture Content (in steam lines) Due to the unknowns related to the relative impact of the influencing factors and the complexities of the interactions between these factors, it is difficult to accurately predict the expected wear rate in a given piping segment in a new plant. This paper provides: - a description of FAC and the factors that influence the FAC degradation rate, - an assessment of the level of FAC resistance of AP1000 R secondary side system piping, - an explanation of options to increase FAC resistance and associated benefits/cost, - discussion of development of a tool for predicting FAC degradation rate in new nuclear power plants. (authors)

  8. Numerical simulation study on the air/water countercurrent flow limitation in nuclear reactors

    Energy Technology Data Exchange (ETDEWEB)

    Morghi, Youssef; Mesquita, Amir Z., E-mail: ssfmorghi@gmail.com, E-mail: amir@cdtn.br [Centro de Desenvolvimento da Tecnologia Nuclear (CDTN/CNEN-MG), Belo Horizonte, MG (Brazil); Puente, Jesus, E-mail: jpuente720@gmail.com [Centro Federal de Educaçao Tecnologica Celso Suckowda Fonseca (CEFET), Angra dos Reis, RJ (Brazil); Baliza, Ana R., E-mail: baliza@eletronuclear.gov.br [Eletrobras Eletronuclear Angra dos Reis, RJ (Brazil)

    2017-07-01

    After a loss-of-coolant accident (LOCA) in a Pressurized Water Reactor (PWR), the temperature of the fuel elements cladding increases dramatically due to the heat produced by the fission products decay, which is not adequately removed by the vapor contained in the core. In order to avoid this sharp rise in temperature and consequent melting of the core, the Emergency Core Cooling System is activated. This system initially injects borated water from accumulator tanks of the reactor through the inlet pipe (cold leg) and the outlet pipe (hot leg), or through the cold leg only, depending on the plant manufacturer. Some manufacturers add to this, direct injection into the upper plenum of the reactor. The penetration of water into the reactor core is a complex thermo fluid dynamic process because it involves the mixing of water with the vapor contained in the reactor, added to that generated in the contact of the water with the still hot surfaces in various geometries. In some critical locations, the vapor flowing in the opposite direction of the water can control the penetration of this into the core. This phenomenon is known as Countercurrent Flow Limitation (CCFL) or Flooding, and it is characterized by the control that a gas exerts in the liquid flow in the opposite direction. This work presents a proposal to use a CFD to simulate the CCFL phenomenon. Numerical computing can provide important information and data that is difficult or expensive to measure or test experimentally. Given the importance of computational science today, it can be considered a third and independent branch of science on an equal footing with the theoretical and experimental sciences. (author)

  9. Continuous flow solvent extraction system for the determination of trace amounts of uranium in nuclear waste reprocessing solutions

    International Nuclear Information System (INIS)

    Atallah, R.H.; Christian, G.D.

    1988-01-01

    A system is described for the continuous flow solvent extraction of uranium using isobutyl methyl ketone (IBMK) in the presence of a salting-out solution. This is used as a pre-step to spectrophotometric determination with 2-(5-bromo-2-pyridylazo)-5-(diethylamino)phenol (Br-PADAP) in methanol. The method utilises an on-line solvent extraction unit, interfaced to a spectrophotometric flow injection analysis (FIA) detection system. The extraction unit can be operated in two ways. In the first, 0.6 ml of a sample was loaded (injected) and then the separated organic phase was mixed directly with the reagent. This permits the handling of 20 samples per hour and a limit of determination of 0.2 p.p.m. of uranium. In the second method, a continuously pumped sample was brought into contact with a fixed volume of the organic phase, which was recirculated in a closed loop. Recirculation results in the extraction of more analyte with the same volume of the organic phase present in the loop. For a sample loading volume of 3 ml, an enrichment factor of 4 was obtained. The determination of uranium at the 50 p.p.b. level can be achieved using this automated liquid - liquid extraction system. Lower concentrations of uranium can also be measured by further increasing the sample volume. This method was tested on a synthetic nuclear waste solution. (author)

  10. A practical system for the measurement of flows in a nuclear reactor

    International Nuclear Information System (INIS)

    Carrion V, F.J.

    1980-01-01

    With the purpose of making practical the neutron flux map in a nuclear reactor, the traditional foils are replaced by calibrated wires, and the activity rates through the wire are maked with a directional detector coupled to a collimator and to a servomechanism that moves the wire, to make the measurements through it. The servomechanism can be automatically controlled, and the information can be stored in a multichannel or directly pass to a plotter according to the continued or step by step measurements through the wire. For the system design it was necessary to fix the collimator dimensions according to the detector and the wire characteristics. In conclusion, this method is practical and exact, and their automation allows time economy. (author)

  11. Experimental investigation on single-phase pressure losses in nuclear debris beds: Identification of flow regimes and effective diameter

    Energy Technology Data Exchange (ETDEWEB)

    Clavier, R., E-mail: remi.clavier@irsn.fr [Institut de Radioprotection et de Sûreté Nucléaire (IRSN) – PSN-RES/SEREX/LE2M, Cadarache bât. 327, 13115 St Paul-lez-Durance (France); Chikhi, N., E-mail: nourdine.chikhi@irsn.fr [Institut de Radioprotection et de Sûreté Nucléaire (IRSN) – PSN-RES/SEREX/LE2M, Cadarache bât. 327, 13115 St Paul-lez-Durance (France); Fichot, F. [Institut de Radioprotection et de Sûreté Nucléaire (IRSN) – PSN-RES/SAG/LEPC, Cadarache bât. 700, 13115 St Paul-lez-Durance (France); Quintard, M. [Université de Toulouse – INPT – UPS – Institut de Mécanique des Fluides de Toulouse (IMFT), Allée Camille Soula, F-31400 Toulouse (France); CNRS – IMFT, F-31400 Toulouse (France)

    2015-10-15

    Highlights: • Single-phase pressure drops versus flow rates in particle beds are measured. • Conditions are representative of the reflooding of a nuclear fuel debris bed. • Darcy, weak inertial, strong inertial and weak turbulent regimes are observed. • A Darcy–Forchheimer law is found to be a good approximation in this domain. • A predictive correlation is derived from new experimental data. - Abstract: During a severe nuclear power plant accident, the degradation of the reactor core can lead to the formation of debris beds. The main accident management procedure consists in injecting water inside the reactor vessel. Nevertheless, large uncertainties remain regarding the coolability of such debris beds. Motivated by the reduction of these uncertainties, experiments have been conducted on the CALIDE facility in order to investigate single-phase pressure losses in representative debris beds. In this paper, these results are presented and analyzed in order to identify a simple single-phase flow pressure loss correlation for debris-bed-like particle beds in reflooding conditions, which cover Darcean to Weakly Turbulent flow regimes. The first part of this work is dedicated to study macro-scale pressure losses generated by debris-bed-like particle beds, i.e., high sphericity (>80%) particle beds with relatively small size dispersion (from 1 mm to 10 mm). A Darcy–Forchheimer law, involving the sum of a linear term and a quadratic deviation, with respect to filtration velocity, has been found to be relevant to describe this behavior in Darcy, Strong Inertial and Weak Turbulent regimes. It has also been observed that, in a restricted domain (Re = 15 to Re = 30) between Darcy and Weak Inertial regimes, deviation is better described by a cubic term, which corresponds to the so-called Weak Inertial regime. The second part of this work aims at identifying expressions for coefficients of linear and quadratic terms in Darcy–Forchheimer law, in order to obtain a

  12. Modelling of thermally driven groundwater flow in a facility for disposal of spent nuclear fuel in deep boreholes

    Energy Technology Data Exchange (ETDEWEB)

    Marsic, Nico; Grundfelt, Bertil [Kemakta Konsult AB, Stockholm (Sweden)

    2013-09-15

    In this report calculations are presented of buoyancy driven groundwater flow caused by the emission of residual heat from spent nuclear fuel deposited in deep boreholes from the ground surface in combination with the natural geothermal gradient. This work has been conducted within SKB's programme for evaluation of alternative methods for final disposal of spent nuclear fuel. The basic safety feature of disposal of spent nuclear fuel in deep boreholes is that the groundwater at great depth has a higher salinity, and hence a higher density, than more superficial groundwater. The result of this is that the deep groundwater becomes virtually stagnant. The study comprises analyses of the effects of different inter-borehole distances as well as the effect of different permeabilities in the backfill and sealing materials in the borehole and of different shapes of the interface between fresh and saline groundwater. The study is an update of a previous study published in 2006. In the present study, the facility design proposed by Sandia National Laboratories has been studied. In this design, steel canisters containing two BWR elements or one PWR element are stacked on top of each other between 3 and 5 kilometres depth. In order to host all spent fuel from the current Swedish nuclear programme, about 80 such holes are needed. The model used in this study comprises nine boreholes spaced 100 metres alternatively 50 metres apart in a 3{Chi}3 matrix. In one set of calculations the salinity in the groundwater was assumed to increase from zero above 700 metres depth to 10% by weight at 1500 metres depth and below. In another set, a sharper salinity gradient was applied in which the salinity increased from 0 to 10% between 1400 and 1500 metres depth. A geothermal gradient of 16 deg C/km was applied. The heat output from the spent fuel was assumed to decrease by time in manner consistent with the radioactive decay in the fuel. When the inter-borehole distance decreased from

  13. A study on nuclear heat load tolerable for NET/TF coils cooled by internal flow of helium II

    International Nuclear Information System (INIS)

    Hofmann, A.

    1988-02-01

    NbTi cables cooled by internal flow of superfluid helium are considered an option for the design of NET/TF coils with about 11 T peak fields. Starting from an available winding cross section of 0.61x0.61 m 2 for a 8 MA turns coil made of a 16 kA conductor it is shown that sufficient hydraulic cross section can be provided within such cables to remove the expected thermal load resulting from nuclear heating with exponential decay from inboard to outboard side of the winding. The concept is a pancake type coil with 1.8 K helium fed-in the high field region of each pancake. The temperature distribution within such coils is calculated, and the local safety margin is determined from temperature and field. The calculation takes account of nuclear and a.c. heating, and of thermal conductance between the individual layers and the coil casing. It is shown that operation with 1.8 K inlet and about 3 K outlet temperature is possible. The electrical insulation with about 0.5 mm thickness proves to provide sufficient thermal insulation. No additional thermal shield is required between the coil casing and the winding package. Two different types of conductors are being considered: a) POLO type cable with quadratic cross section and a central circular coolant duct, and b) an LCT type cable with two conductors wound in hand. Both concepts with about 500 m length of the cooland channels are shown to meet the requirements resulting from a peak nuclear heat load of 0.3 mW/cm 3 in the inboard turns. The hydraulic diameters are sufficient to operate each coils with self-sustained fountain effect pumps. Even appreciably higher heat loads with up to 3 mW/cm 3 of nuclear heating can be tolerated for the POLO type cable when the hydraulic diameter is enlarged to its maximum of 17 mm. (orig.) [de

  14. Analysis of flow accelerated corrosion in piping of nuclear power plants

    International Nuclear Information System (INIS)

    Naitoh, Masanori; Uchida, Shunsuke; Uehara, Yasushi; Koshizuka, Seiichi

    2008-01-01

    The FAC evaluation models and procedures were developed by coupling corrosion models and flow dynamics models. The newly developed wall thinning calculation code coupled with the static electrochemistry model and the dynamic oxide film growth model was validated by comparison with measurements. As one of countermeasures for mitigation of FAC for the secondary cooling system of PWRs, a location of hydrazine injection as a scavenger of oxygen was discussed from the viewpoint of water chemistry improvement. The FAC dangerous zone was defined as overlapping conditions of ranges of influential parameters for FAC occurrence. When hydrazine was injected at the deaerator inlet, a FAC dangerous zone did not appear at all and oxygen concentration was kept sufficiently low level to prevent corrosion of Inconel tubes of steam generators. From a view point of mitigation of FAC, a small addition of oxygen was recommended. (author)

  15. Two new nuclear isolation buffers for plant DNA flow cytometry: A test with 37 species

    Czech Academy of Sciences Publication Activity Database

    Loureiro, J.; Rodriguez, E.; Doležel, Jaroslav; Santos, C.

    2007-01-01

    Roč. 100, č. 4 (2007), s. 875-888 ISSN 0305-7364 R&D Projects: GA ČR GA521/06/1723; GA ČR(CZ) GA521/05/0257; GA MŠk(CZ) LC06004 Grant - others:Mendelova zemědělská a lesnická univerzita v Brně / Agronomická fakulta(CZ) ME 844 Institutional research plan: CEZ:AV0Z50380511 Source of funding: V - iné verejné zdroje ; V - iné verejné zdroje ; V - iné verejné zdroje ; V - iné verejné zdroje Keywords : flow cytometry * general purpose buffer * genome size Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.939, year: 2007

  16. 3D Numerical study of the external flow effect on the heat transfer in a radiometric calorimeter dedicated to nuclear heating measurements

    International Nuclear Information System (INIS)

    Muraglia, M.; Reynard-Carette, C.; Brun, J.; Carette, M.; Lyoussi, A.

    2013-06-01

    Improvement of measurements in reactor is still a challenge. Thus, this work focuses on numerical studies of one sensor dedicated to nuclear heating measurements: a radiometric complex calorimeter. More precisely, using a simplified conduction heat model, this work presents the first full 3D simulations of a simplified calorimeter reduced to the complex calorimeter head showing that the key parameter for the sensitivity control is the convective heat transfers between the calorimeter and its external surrounding. The effect of external flow velocity on the calorimeter head response is determined for different flow regimes (natural convection, forced convection) and numerical results are found to be in agreement with experimental results under non-irradiated conditions obtained for the complex calorimeter. Moreover, in order to understand and describe fully the mechanisms leading at the different calorimeter heat transfer, the flow velocity dynamics should be added in the model. In a first approach, due to low influence of the flow velocity for tested power range, a static cooling fluid around the calorimeter head is added in the model. Then, in order to get the full flow dynamics, using Boussinesq approximation, a new 2D fluid model, including both temperature field and flow velocity dynamics, is derived taking into account the nuclear heating effect on the flow. (authors)

  17. Uranium analysis in water flowing by the nuclear track detection method on solid dielectric materials

    International Nuclear Information System (INIS)

    Arambula, H.

    1981-01-01

    The objective of this experiment was threefold: to study the content of uranium in tap and spring water, to establish a technique for the quantitative analysis for the presence of uranium in liquids, and to test the qualities as detector fission fragments of three solid insulator materials using the nuclear tracks register method. The latter allows for the measurement of concentrations of fissile elements up to 10 -12 gr/gm employing (n, f.f.) reactions. The test samples were of tap water and of water from six fresh water springs located in San Luis Potosi and Guanajuato. Glass, lexan polycarbonate and muscovite mica were the detector materials used. The technique consisted in evaporating the water from the test samples, which had been previously placed upon the detector materials, and in doing the same for the standard control sample solutions having known concentrations of uranium. All the samples were then irradiated with thermal neutrons, and the 235 U, present in the samples, fissioned. The fission fragments produced permanent damage on the detectors, known as latent tracks. A specific corroding chemical was then applied to each detector which caused the latent tracks to dissolve into grooves. Known as etching tracks, these grooves were microscopically visible and could be measured for track density (tracks/mm 2 ). The concentrations of uranium present in the test samples were measured by comparing the track densities of the test samples with those of the standard control samples. The concentration of uranium found in the spring water samples ranged from 0.09 to 0.89 μqr.U/1, and those of tap water, from 0.18 to 0.19 μqr U/1. Lexan polycarbonate and muscovite mica proved to be better, as detectors, than glass. Glass for quantitative analysis, we found not recommendable as a detector material because of its alterable composition in the presence of uranium. (author)

  18. Improvement of Diagnostic Flow Chart in Severe Accident Management Guidance for Nuclear Power Plants in Korea

    International Nuclear Information System (INIS)

    Jang, So Won; Lee, Su Won; Oh, Hae Cheol

    2016-01-01

    Development of generic SAMG in the United States was undertaken by WOG, Combustion Engineering Owner’s Group (CEOG), and Babcock and Wilcox Owner’s Group (BWOG) in the 1990s. They made efforts to develop generic SAMG specific to the individual plant designs to satisfy the regulatory concerns for severe accident managements. Recently, the SAMG of the Pressurized Water Reactor Owner's Group (PWROG) which is applicable to Babcock and Wilcox (B and W), Combustion Engineering (CE), and Westinghouse Pressurizer Water Reactor (PWR) Nuclear Steam Supply System (NSSS) designs was developed in February 2016 by incorporating the best features from the previous PWR generic SAMG. However, the structure of the PWROG SAMG is also based on the WOG SAMG. The purpose of this study is to provide improvements of the Korean SAMG by comparing between the Korean SAMG and the PWROG SAMG to reflect the state of the art and trends. The improved DFC for the Korean SAMG is proposed by comparing between the Korean SAMG and the PWROG SAMG. If the improved DFC is adopted, the level to identify the severity of the plant condition is diversified, the order of priority of the strategies is changed, the entry condition into the RCS injection strategy is changed, LTMG-01and SAEG-01 come under the DFC as parts of the step, and the step whether CCI is occurred or not is added in DFC. In order to increase the effectiveness of the Korean SAMG and to reflect the state of the art and trends, it is appropriate to apply the suggestions contained in this paper to the Korean SAMG

  19. Flow cytometry in the diagnosis of myelodysplastic syndromes (MDS) and the value of myeloid nuclear differentiation antigen (MNDA).

    Science.gov (United States)

    Bellos, Frauke; Kern, Wolfgang

    2014-09-25

    Background: Confirming diagnosis of myelodysplastic syndromes (MDS) is often challenging. Standard diagnostic methods are cytomorphology (CM) and cytogenetics (CG). Multiparameter flow cytometry (MFC) is upcoming in MDS diagnostic work up, comparability and investigator experience are critical. Myeloid nuclear differentiation antigen (MNDA) in myelomonocytic cells might be expressed more weakly in patients with MDS. The analysis of MNDA may thus improve diagnostic capabilities of MFC in MDS. Methods: Staining methods and antibody combinations for MFC in MDS are outlined, giving details for interpretation of results in regard to dyspoiesis. MFC results are correlated with CM and CG and with survival data. Use of myeloid nuclear differentiation antigen (MNDA) in MDS diagnostics was evaluated in 239 patients with MDS, AML, other cytopenic conditions and in 30 negative controls. Results: Strong correlation between findings in CM and MFC was found; MFC results correlated well with those of CG. Patients with higher grades of dysplasia in MFC had shorter overall survival. Percentages of granulocytes and monocytes with diminished MNDA expression (%dimG, %dimM) were higher in patients with MDS and AML. Mean fluorescence intensity (MFI) of MNDA in monocytes was lower in MDS and AML. Cut-off values for %dimG (12%) and %dimM (22%) as well as for MFI in monocytes (72) were defined discriminating between MDS and non-MDS. Conclusion: MFC adds significant information on dyspoiesis in the diagnostic work up for MDS and provides prognostic information. MNDA expression can be assessed by MFC and may facilitate evaluation of dyspoiesis when added to MDS MFC panels. © 2014 Clinical Cytometry Society. Copyright © 2014 Clinical Cytometry Society.

  20. A study on quantification of the information flow and effectiveness of information aids for diagnosis tasks in nuclear power plants

    International Nuclear Information System (INIS)

    Kim, Jong Hyun

    2004-02-01

    Diagnosis is one of the most complex and mental resource-demanding tasks in nuclear power plants (NPPs), especially, to main control room (MCR) operators. Diagnosis is a crucial part of disturbance control in NPPs, since it is a prerequisite task for initiating operating procedures. In order to design a control room feature for NPPs, three elements need to be considered: 1) the operational tasks that must be performed, 2) a model of human performance for these tasks, and 3) a model of how control room features are intended to support performance. The operational tasks define the classes of performance that must be considered. A model of human performance makes more explicit the requirements for accurate and efficient performance and reveals potential sources of error. Finally, the model of support allows the generation of specific hypotheses about how performance is facilitated in the control room. The model of support needs to be developed based on the human performance model. This paper proposes three approaches for the system design of operator support systems to aid MCR operators' diagnosis tasks in NPPs, considering the above three elements. This paper presents 1) a quantitative approach to modeling the information flow of diagnosis tasks, 2) strategy-based evaluation of information aids for diagnosis tasks, and 3) quantitative evaluation of NPP decision support systems. As an analysis of diagnosis tasks, this paper presents a method to quantify the cognitive information flow of diagnosis tasks, integrating a stage model (a qualitative approach) with information theory (a quantitative approach). The method includes: 1) constructing the information flow model, which consists of four stages based on operating procedures of NPPs: and 2) quantifying the information flow using Conant's model, a kind of information theory. Then, three experiments were conducted to evaluate the effectiveness of the proposed approach to predicting human performances

  1. Seed gene flow and fine-scale structure in a Mediterranean pine ( Pinus pinaster Ait.) using nuclear microsatellite markers.

    Science.gov (United States)

    González-Martínez, C.; Gerber, S.; Cervera, T.; Martínez-Zapater, M.; Gil, L.; Alía, R.

    2002-06-01

    The Mediterranean populations of maritime pine ( Pinus pinaster Ait.) are typically small and have a scattered distribution, being threatened by human activities and forest fires. In the framework of the genetic-resources conservation program of this species, a native multi-age stand located in a Mediterranean area (central Spain) was studied using three highly polymorphic nuclear microsatellites (SSRs). Spatial autocorrelation analysis was conducted using Moran's index in order to detect fine-scale structure in both natural regeneration and mature trees. The spatial pattern of seed flow based on dispersed progeny was studied using a highly reliable subset of parent-offspring matches obtained by means of parentage analysis and simulation-based calculation of statistical confidence. Maritime pine showed a fine-scale structure at the seedling stage. In natural regeneration, the autocorrelograms indicated a patch size of approximately 10 m. The fine-scale structure seems to be produced by a restricted seed gene flow. In fact, there was an excess of parent-offspring matches in a radius of 15 m from the parent trees. Pines with a heavy seed, such as P. pinaster, are expected to have a short dispersal distance, thus producing a fine-scale structure. However, the fine-scale structure did not persist in the mature trees. Within-population genetic structure in Mediterranean pines may be affected by a number of post-dispersal events (e.g. mortality due to the severity of the Mediterranean climate and animal-mediated secondary dispersal during the summer period). Thus, great alteration in the pattern produced by the initial seed rain and differences in genetic structure between tree cohorts are expected.

  2. Challenges of setting up flow cytometry for diagnosis of leukemia ...

    African Journals Online (AJOL)

    The bone marrow (BM) is a complex tissue containing cells of multiple hematopoietic cell lineages in all stages of development. Flow cytometric immunophenotyping evaluates the frequencies of the various leukocyte (sub) populations in BM and blood that then helps in the diagnosis of leukemia's. The aim of this study was ...

  3. Transient core characteristics of small molten salt reactor coupling problem between heat transfer/flow and nuclear fission reaction

    International Nuclear Information System (INIS)

    Yamamoto, Takahisa; Mitachi, Koshi

    2004-01-01

    This paper performed the transient core analysis of a small Molten Salt Reactor (MSR). The emphasis is that the numerical model employed in this paper takes into account the interaction among fuel salt flow, nuclear reaction and heat transfer. The model consists of two group diffusion equations for fast and thermal neutron fluexs, balance equations for six-group delayed neutron precursors and energy conservation equations for fuel salt and graphite moderator. The results of transient analysis are that (1) fission reaction (heat generation) rate significantly increases soon after step reactivity insertion, e.g., the peak of fission reaction rate achieves about 2.7 times larger than the rated power 350 MW when the reactivity of 0.15% Δk/k 0 is inserted to the rated state, and (2) the self-control performance of the small MSR effectively works under the step reactivity insertion of 0.56% Δk/k 0 , putting the fission reaction rate back on the rated state. (author)

  4. Ground-water flow near two radioactive-waste-disposal areas at the Western New York Nuclear Service Center, Cattaraugus County, New York; results of flow simulation

    Science.gov (United States)

    Bergeron, M.P.; Bugliosi, E.F.

    1988-01-01

    Two adjacent burial areas were excavated in a clay-rich till at a radioactive waste disposal site near West Valley in Cattaraugus County, N.Y.: (1) which contains mainly low-level radioactive wastes generated onsite by a nuclear fuel reprocessing plant, has been in operation since 1966; and (2) which contains commercial low-level radioactive wastes, was operated during 1963-75. Groundwater below the upper 3 meters of till generally moves downward through a 20- to 30-meter thick sequence of tills underlain by lacustrine and kame-delta deposits of fine sand and silt. Groundwater in the weathered, upper 3 meters of till can move laterally for several meters before either moving downward into the kame-delta deposits or discharging to the land surface. A two-dimensional finite-element model that simulates two vertical sections was used to evaluate hydrologic factors that control groundwater flow in the till. Conditions observed during March 1983 were reproduced accurately in steady-state simulations that used four isotropic units of differing hydraulic conductivity to represent two fractured and weathered till units near land surfaces, an intermediate group of isolated till zones that contain significant amounts of fine sand and silt, and a sequence of till units at depths that have been consolidated by overburden pressure. Recharge rates used in the best-fit simulation ranged from 1.4 cm/yr along smooth, sloping or compacted surfaces to 3.8 cm/yr near swampy areas. Values of hydraulic conductivity and infiltration used in the calibrated best-fit model were nearly identical to values used in a previous model analysis of the nearby commercial-waste burial area. Results of the model simulations of a burial pit assumed to be filled with water indicate that water near the bottom of the burial pit would migrate laterally in the shallow, weathered till for 5 to 6 meters before moving downward into the unweathered till, and water near the top of the pit would move laterally

  5. Thermal and flow analysis of the Fluor Daniel, Inc., Nuclear Material Storage Facility renovation design (initial 30% effort of Title 1)

    International Nuclear Information System (INIS)

    Steinke, R.G.; Mueller, C.; Knight, T.D.

    1998-03-01

    The computational fluid dynamics code CFX4.2 was used to evaluate steady-state thermal-hydraulic conditions in the Fluor Daniel, Inc., Nuclear Material Storage Facility renovation design (initial 30% of Title 1). Thirteen facility cases were evaluated with varying temperature dependence, drywell-array heat-source magnitude and distribution, location of the inlet tower, and no-flow curtains in the drywell-array vault. Four cases of a detailed model of the inlet-tower top fixture were evaluated to show the effect of the canopy-cruciform fixture design on the air pressure and flow distributions

  6. Assessment of Equine Autoimmune Thrombocytopenia (EAT by flow cytometry

    Directory of Open Access Journals (Sweden)

    Schwarzwald Colin

    2001-04-01

    Full Text Available Abstract Rationale Thrombocytopenia is a platelet associated process that occurs in human and animals as result of i decreased production; ii increased utilization; iii increased destruction coupled to the presence of antibodies, within a process know as immune-mediated thrombocytopenia (IMT; or iv platelet sequestration. Thus, the differentiation of the origin of IMT and the development of reliable diagnostic approaches and methodologies are important in the clarification of IMT pathogenesis. Therefore, there is a growing need in the field for easy to perform assays for assessing platelet morphological characteristics paired with detection of platelet-bound IgG. Objectives This study is aimed to develop and characterize a single color flow cytometric assay for detection of platelet-bound IgG in horses, in combination with flow cytometric assessment of platelet morphological characteristics. Findings The FSC and SSC evaluation of the platelets obtained from the thrombocytopenic animals shows several distinctive features in comparison to the flow cytometric profile of platelets from healthy animals. The thrombocytopenic animals displayed i increased number of platelets with high FSC and high SSC, ii a significant number of those gigantic platelets had strong fluorescent signal (IgG bound, iii very small platelets or platelet derived microparticles were found significantly enhanced in one of the thrombocytopenic horses, iv significant numbers of these microplatelet/microparticles/platelet-fragments still carry very high fluorescence. Conclusions This study describes the development and characterization of an easy to perform, inexpensive, and noninvasive single color flow cytometric assay for detection of platelet-bound IgG, in combination with flow cytometric assessment of platelet morphological characteristics in horses.

  7. Non-equilibrium estimates of gene flow inferred from nuclear genealogies suggest that Iberian and North African wall lizards (Podarcis spp. are an assemblage of incipient species

    Directory of Open Access Journals (Sweden)

    Harris D James

    2008-02-01

    Full Text Available Abstract Background The study of recently-diverged species offers significant challenges both in the definition of evolutionary entities and in the estimation of gene flow among them. Iberian and North African wall lizards (Podarcis constitute a cryptic species complex for which previous assessments of mitochondrial DNA (mtDNA and allozyme variation are concordant in describing the existence of several highly differentiated evolutionary units. However, these studies report important differences suggesting the occurrence of gene flow among forms. Here we study sequence variation in two nuclear introns, β-fibint7 and 6-Pgdint7, to further investigate overall evolutionary dynamics and test hypotheses related to species delimitation within this complex. Results Both nuclear gene genealogies fail to define species as monophyletic. To discriminate between the effects of incomplete lineage sorting and gene flow in setting this pattern, we estimated migration rates among species using both FST-based estimators of gene flow, which assume migration-drift equilibrium, and a coalescent approach based on a model of divergence with gene flow. Equilibrium estimates of gene flow suggest widespread introgression between species, but coalescent estimates describe virtually zero admixture between most (but not all species pairs. This suggests that although gene flow among forms may have occurred the main cause for species polyphyly is incomplete lineage sorting, implying that most forms have been isolated since their divergence. This observation is therefore in accordance with previous reports of strong differentiation based on mtDNA and allozyme data. Conclusion These results corroborate most forms of Iberian and North African Podarcis as differentiated, although incipient, species, supporting a gradual view of speciation, according to which species may persist as distinct despite some permeability to genetic exchange and without having clearly definable genetic

  8. Genome size estimations on Ulmus minor Mill., Ulmus glabra Huds., and Celtis australis L. using flow cytometry.

    Science.gov (United States)

    Loureiro, J; Rodriguez, E; Gomes, A; Santos, C

    2007-07-01

    The Ulmaceae family is composed of nearly 2000 species widely distributed in the northern hemisphere. Despite their wide distribution area, there are only four native species in the Iberian Peninsula. In this work the genome size of three of those species (ULMUS MINOR, U. GLABRA, and CELTIS AUSTRALIS) was estimated using flow cytometry. The nuclear DNA content of C. AUSTRALIS was estimated as 2.46 +/- 0.061 pg/2C, of U. MINOR as 4.25 +/- 0.158 pg/2C, and of U. GLABRA as 4.37 +/- 0.103 pg/2C of DNA. No statistically significant differences were detected among individuals of the same species. These species revealed to be problematic for flow cytometric analyses, due to the release of mucilaginous compounds into the nuclear suspension. Despite that, the modified protocol here presented ensured high quality analyses (low coefficient of variation and background debris and nuclear fluorescence stability), opening good perspectives on its application to estimate the genome size of species with similar problems.

  9. A Validation Process for the Groundwater Flow and Transport Model of the Faultless Nuclear Test at Central Nevada Test Area

    Energy Technology Data Exchange (ETDEWEB)

    Ahmed Hassan

    2003-01-01

    Many sites of groundwater contamination rely heavily on complex numerical models of flow and transport to develop closure plans. This has created a need for tools and approaches that can be used to build confidence in model predictions and make it apparent to regulators, policy makers, and the public that these models are sufficient for decision making. This confidence building is a long-term iterative process and it is this process that should be termed ''model validation.'' Model validation is a process not an end result. That is, the process of model validation cannot always assure acceptable prediction or quality of the model. Rather, it provides safeguard against faulty models or inadequately developed and tested models. Therefore, development of a systematic approach for evaluating and validating subsurface predictive models and guiding field activities for data collection and long-term monitoring is strongly needed. This report presents a review of model validation studies that pertain to groundwater flow and transport modeling. Definitions, literature debates, previously proposed validation strategies, and conferences and symposia that focused on subsurface model validation are reviewed and discussed. The review is general in nature, but the focus of the discussion is on site-specific, predictive groundwater models that are used for making decisions regarding remediation activities and site closure. An attempt is made to compile most of the published studies on groundwater model validation and assemble what has been proposed or used for validating subsurface models. The aim is to provide a reasonable starting point to aid the development of the validation plan for the groundwater flow and transport model of the Faultless nuclear test conducted at the Central Nevada Test Area (CNTA). The review of previous studies on model validation shows that there does not exist a set of specific procedures and tests that can be easily adapted and

  10. Security of supply and line flow following the shut-down of nuclear power plants in Germany. Have shortages to be expected?

    International Nuclear Information System (INIS)

    Kunz, Friedrich; Moest, Dominik

    2011-01-01

    This paper, which examines the impacts of phasing out nuclear power in Germany, is the first to include an analysis of energy supply security and critical line flows in both the German and Central European electricity networks. The technical-economic model of the European electricity market, ELMOD, is used to simulate alternative power plant dispatch, imports, exports, and network use for a representative winter day. The results suggest that the shutdown of Germany's nuclear plants will result in higher net imports, especially from the Netherlands, Austria, and Poland, and that electricity generation from fossil fuels will increase slightly in Germany and in Central Europe. We find that no additional imports will come from nuclear plants since they are already fully utilized in the merit order, and that electricity prices will rise on average by a few Euros per MWh. We conclude that closing the seven nuclear power plants within the government's moratorium will cause no significant supply security issues or network constraints and an eventual full phase-out seem to be possible due to the completion of several new conventional power plants now under construction. Finally, we suggest that a nuclear phase-out in Germany within the next 3-7 years will not undermine security of supply and network stability in Germany and Central Europe.

  11. Cytometric comparisons between circulating tumor cells from prostate cancer patients and the prostate-tumor-derived LNCaP cell line

    Science.gov (United States)

    Lazar, Daniel C.; Cho, Edward H.; Luttgen, Madelyn S.; Metzner, Thomas J.; Loressa Uson, Maria; Torrey, Melissa; Gross, Mitchell E.; Kuhn, Peter

    2012-02-01

    Many important experiments in cancer research are initiated with cell line data analysis due to the ease of accessibility and utilization. Recently, the ability to capture and characterize circulating tumor cells (CTCs) has become more prevalent in the research setting. This ability to detect, isolate and analyze CTCs allows us to directly compare specific protein expression levels found in patient CTCs to cell lines. In this study, we use immunocytochemistry to compare the protein expression levels of total cytokeratin (CK) and androgen receptor (AR) in CTCs and cell lines from patients with prostate cancer to determine what translational insights might be gained through the use of cell line data. A non-enrichment CTC detection assay enables us to compare cytometric features and relative expression levels of CK and AR by indirect immunofluorescence from prostate cancer patients against the prostate cancer cell line LNCaP. We measured physical characteristics of these two groups and observed significant differences in cell size, fluorescence intensity and nuclear to cytoplasmic ratio. We hope that these experiments will initiate a foundation to allow cell line data to be compared against characteristics of primary cells from patients.

  12. Improved graft survival in highly sensitized patients undergoing renal transplantation after the introduction of a clinically validated flow cytometry crossmatch.

    LENUS (Irish Health Repository)

    Limaye, Sandhya

    2009-04-15

    Flow cytometric techniques are increasingly used in pretransplant crossmatching, although there remains debate regarding the clinical significance and predictive value of donor-specific antibodies detected by flow cytometry. At least some of the discrepancies between published studies may arise from differences in cutoffs used and lack of standardization of the test.

  13. The post irradiation examination of fuel in support of Bruce A nuclear division fueling with flow program

    International Nuclear Information System (INIS)

    Montin, J.; Sagat, S.; Day, R.; Novak, J.; Bromfield, H.

    1995-01-01

    Bruce A Nuclear Division (BAND) units are operating at ∼ 75% of full power, because of the potential of a power pulse in the event of an inlet header break. As a result, BAND is converting to fueling with flow, to eliminate the potential of a power pulse and to allow for full-power operation. Concerns regarding the integrity of the end-of-life (EOL) bundles interacting with the latch at the downstream end of the fuel channel were raised. BAND carried out a test program in which EOL bundles in the upstream position 13 of Unit 2 were cascaded into the downstream latch position 1 of another channel. Six of twelve cascaded bundles and two typical EOL position 13 (benchmark) bundles were selected for post-irradiation examination (PIE). Incipient cracks were found in the assembly welds (endplateto-endcap welds) of all six cascaded bundles. No incipient cracks were found in the benchmark bundles. Metallographic and fractographic examination, along with crack dating, and hydrogen and deuterium analyses, indicated that the incipient cracks were the result of delayed-hydride assisted cracking at the EOL. Consequently, Ontario Hydro changed the design of the outlet shield plug to support all three rings of the fuel bundle, to minimize stress and prevent endplate cracking. Also, an ultrasonic endplate inspection tool (UT) was developed and located in the fuel bay. to inspect fuelbundle endplates for cracks. A second test was done involving a series of four bundle cascades in BAND Unit 4 channels that had new outlet shield plugs. The latch bundles were discharged after a hot shutdown. The cascaded Unit 2 and Unit 4 latch bundles were checked for cracks using the UT. The PIE found incipient cracks or less-than-ideal welds in the assembly welds of fuel elements from Unit 2 (latch-supported fuel bundles) that had been identified by the UT as having incipient cracks. No incipient cracks were found in the assembly welds of fuel elements from Unit 4 (new outlet shield

  14. The post-irradiation examination of fuel in support of Bruce A Nuclear Division fueling with flow program

    International Nuclear Information System (INIS)

    Montin, J.; Sagat, S.

    1995-10-01

    Bruce A Nuclear Division (BAND) units are operating at ∼ 75% of full power, because of the potential of a power pulse in the event of an inlet header break. As a result, BAND is converting to fueling with flow, to eliminate the potential of a power pulse and to allow for full-power operation. Concerns regarding the integrity of the end-of-life (EOL) bundles interacting with the latch at the downstream end of the fuel channel were raised. BAND carried out a test program in which EOL bundles in the upstream position of 13 of Unit 2 were cascaded into the downstream latch position 1 of another channel. Six of twelve cascaded bundles and two typical EOL position 13 (benchmark) bundles were selected for post-irradiation examination (PIE). Incipient cracks were found in the benchmark bundles. Metallographic and fractographic examination, along with crack dating, and hydrogen and deuterium analyses, indicated that the incipient cracks were the result of delayed-hydride assisted cracking at the EOL. Consequently, Ontario Hydro changed the design of the outlet shield plug to support all three rings of the fuel bundle, to minimize stress and prevent end plate cracking. Also, an ultrasonic end plate inspection tool (UT) was developed and located in the fuel bay, to inspect fuel-bundle end plates for cracks. A second test was done involving a series of four bundle cascades in BAND Unit 4 channels that had new outlet shield plugs. The latch bundles were discharged after a hot shutdown. The cascaded Unite 2 and Unit 4 latch bundles were checked for cracks using the UT. The PIE found incipient cracks or less-than-ideal welds in the assembly welds of fuel elements from Unit 2 (latch-supported fuel bundles) that had been identified by the UT as having incipient cracks. No incipient cracks were found in the assemble welds of fuel elements from Unit 4 (new outlet shield-supported fuel bundles) confirming the UT results. (author). 5 refs., 8 figs

  15. Identification and detection of murine leukemia blasts by flow cytometry

    OpenAIRE

    sprotocols

    2015-01-01

    Human leukemia has been determined and classified with the help of flow cytometry for the past two decades. Past attempts to detect leukemia blasts relied on both forward and side scatter (FSC and SSC) based on cell size and granularity. However, this technique failed to show a clean separation of blasts from normal lineage cells. In 1993, Borowitz, et al developed flow cytometric analysis to distinguish human leukemia blasts from other normal lineage cells by using fluorescence-conjugated CD...

  16. Expression of Cyclins A, E and Topoisomerase II α correlates with centrosome amplification and genomic instability and influences the reliability of cytometric S-phase determination

    Directory of Open Access Journals (Sweden)

    Laytragoon-Lewin Nongnit

    2003-07-01

    Full Text Available Abstract Background The progression of normal cells through the cell cycle is meticulously regulated by checkpoints guaranteeing the exact replication of the genome during S-phase and its equal division at mitosis. A prerequisite for this achievement is synchronized DNA-replication and centrosome duplication. In this context the expression of cyclins A and E has been shown to play a principal role. Results Our results demonstrated a correlation between centrosome amplification, cell cycle fidelity and the level of mRNA and protein expression of cyclins A and E during the part of the cell cycle defined as G1-phase by means of DNA content based histogram analysis. It is shown that the normal diploid breast cell line HTB-125, the genomically relatively stable aneuploid breast cancer cell line MCF-7, and the genomically unstable aneuploid breast cancer cell line MDA-231 differ remarkably concerning both mRNA and protein expression of the two cyclins during G1-phase. In MDA-231 cells the expression of e.g. cyclin A mRNA was found to be ten times higher than in MCF-7 cells and about 500 times higher than in HTB-125 cells. Topoisomerase II α showed high mRNA expression in MDA compared to MCF-7 cells, but the difference in protein expression was small. Furthermore, we measured centrosome aberrations in 8.4% of the MDA-231 cells, and in only 1.3% of the more stable aneuploid cell line MCF-7. MDA cells showed 27% more incorporation of BrdU than reflected by S-phase determination with flow cytometric DNA content analysis, whereas these values were found to be of the same size in both HTB-125 and MCF-7 cells. Conclusions Our data indicate that the breast cancer cell lines MCF-7 and MDA-231, although both DNA-aneuploid, differ significantly regarding the degree of cell cycle disturbance and centrosome aberrations, which partly could explain the different genomic stability of the two cell lines. The results also question the reliability of cytometric DNA

  17. Flow cytometry of duodenal intraepithelial lymphocytes improves diagnosis of celiac disease in difficult cases.

    Science.gov (United States)

    Valle, Julio; Morgado, José Mario T; Ruiz-Martín, Juan; Guardiola, Antonio; Lopes-Nogueras, Miriam; García-Vela, Almudena; Martín-Sacristán, Beatriz; Sánchez-Muñoz, Laura

    2017-10-01

    Diagnosis of celiac disease is difficult when the combined results of serology and histology are inconclusive. Studies using flow cytometry of intraepithelial lymphocytes (IELs) have found that celiac patients have increased numbers of γδ IELs, along with a decrease in CD3-CD103 + IELs. The objective of this article is to assess the role of flow cytometric analysis of IELs in the diagnosis of celiac disease in difficult cases. A total of 312 patients with suspicion of celiac disease were included in the study. Duodenal biopsy samples were used for histological assessment and for flow cytometric analysis of IELs. In 46 out of 312 cases (14.7%) the combination of serology and histology did not allow the confirmation or exclusion of celiac disease. HLA typing had been performed in 42 of these difficult cases. Taking into account HLA typing and the response to a gluten-free diet, celiac disease was excluded in 30 of these cases and confirmed in the remaining 12. Flow cytometric analysis of IELs allowed a correct diagnosis in 39 out of 42 difficult cases (92.8%) and had a sensitivity of 91.7% (95% CI: 61.5% to 99.8%) and a specificity of 93.3% (95% CI: 77.9% to 99.2%) for the diagnosis of celiac disease in this setting. Flow cytometric analysis of IELs is useful for the diagnosis of celiac disease in difficult cases.

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

    Energy Technology Data Exchange (ETDEWEB)

    Yishai, Yitzhak; Fixler, Dror; Cohen-Kashi, Meir; Zurgil, Naomi; Deutsch, Mordechai [The Biophysical Interdisciplinary Jerome Schottenstein Center for the Research and the Technology of the Cellome, Department of Physics, Bar-Ilan University, Ramat-Gan 52900 (Israel)

    2003-08-07

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

  19. Functional and cytometric examination of different human lung epithelial cell types as drug transport barriers.

    Science.gov (United States)

    Min, Kyoung Ah; Rosania, Gus R; Kim, Chong-Kook; Shin, Meong Cheol

    2016-03-01

    To develop inhaled medications, various cell culture models have been used to examine the transcellular transport or cellular uptake properties of small molecules. For the reproducible high throughput screening of the inhaled drug candidates, a further verification of cell architectures as drug transport barriers can contribute to establishing appropriate in vitro cell models. In the present study, side-by-side experiments were performed to compare the structure and transport function of three lung epithelial cells (Calu-3, normal human bronchial primary cells (NHBE), and NL-20). The cells were cultured on the nucleopore membranes in the air-liquid interface (ALI) culture conditions, with cell culture medium in the basolateral side only, starting from day 1. In transport assays, paracellular transport across all three types of cells appeared to be markedly different with the NHBE or Calu-3 cells, showing low paracellular permeability and high TEER values, while the NL-20 cells showed high paracellular permeability and low TEER. Quantitative image analysis of the confocal microscope sections further confirmed that the Calu-3 cells formed intact cell monolayers in contrast to the NHBE and NL-20 cells with multilayers. Among three lung epithelial cell types, the Calu-3 cell cultures under the ALI condition showed optimal cytometric features for mimicking the biophysical characteristics of in vivo airway epithelium. Therefore, the Calu-3 cell monolayers could be used as functional cell barriers for the lung-targeted drug transport studies.

  20. Application of TEMPPC code to the IEA-R1 nuclear reactor core hydrothermal calculations operating at 2 MW for determining the minimal coolant flow

    International Nuclear Information System (INIS)

    Frajndlich, R.; Sousa, J.A. de.

    1985-01-01

    A thermohydraulic study of the IEA-R1 nuclear reactor core on steady-state operating condition and forced convection, is presented. The objective of this calculation is to obtain the minimal flow rate of coolant necessary at the reactor core, limited by the temperature associated to the beginning of nucleate boiling over the fuel plates at a normal operating power (2MW) for a certain inlet coolant temperature. The coolant system safety level is also calculated in this paper, which is divided in three steps: thermohydraulic calculation, without using the uncertainty factors and, after that, considering these factor by two methods: the statistical and the conventional ones. Whichever the method accepted, the results obtained by the program TEMPPC show a great safety margin with respect to the termohydraulic parameters from the IEA-R1 nuclear reactor. (Author) [pt

  1. Contribution to numerical methods for all Mach flow regimes and to fluid-porous coupling for the simulation of homogeneous two-phase flows in nuclear reactors

    International Nuclear Information System (INIS)

    Zaza, Chady

    2015-01-01

    The numerical simulation of steam generators of pressurized water reactors is a complex problem, involving different flow regimes and a wide range of length and time scales. An accidental scenario may be associated with very fast variations of the flow with an important Mach number. In contrast in the nominal regime the flow may be stationary, at low Mach number. Moreover whatever the regime under consideration, the array of U-tubes is modelled by a porous medium in order to avoid taking into account the complex geometry of the steam generator, which entails the issue of the coupling conditions at the interface with the free-fluid. We propose a new pressure-correction scheme for cell-centered finite volumes for solving the compressible Navier-Stokes and Euler equations at all Mach number. The existence of a discrete solution, the consistency of the scheme in the Lax sense and the positivity of the internal energy were proved. Then the scheme was extended to the homogeneous two-phase flow models of the GENEPI code developed at CEA. Lastly a multigrid-AMR algorithm was adapted for using our pressure-correction scheme on adaptive grids. Regarding the second issue addressed in this work, the numerical simulation of a fluid flow over a porous bed involves very different length scales. Macroscopic interface models - such as Ochoa-Tapia-Whitaker or Beavers-Joseph law for a viscous flow - represent the transition region between the free-fluid and the porous region by an interface of discontinuity associated with specific transmission conditions. An extension to the Beavers-Joseph law was proposed for the convective regime. By introducing a jump in the kinetic energy at the interface, we recover an interface condition close to the Beavers-Joseph law but with a non-linear slip coefficient, which depends on the free-fluid velocity at the interface and on the Darcy velocity. The validity of this new transmission condition was assessed with direct numerical simulations at

  2. Optimization of Cooling Water Flow Rate in Nuclear and Thermal Power Plants Based on a Mathematical Model of Cooling Systems1

    International Nuclear Information System (INIS)

    Murav’ev, V. P.; Kochetkov, A. V.; Glazova, E. G.

    2016-01-01

    A mathematical model and algorithms are proposed for automatic calculation of the optimum flow rate of cooling water in nuclear and thermal power plants with cooling systems of arbitrary complexity. An unlimited number of configuration and design variants are assumed with the possibility of obtaining a result for any computational time interval, from monthly to hourly. The structural solutions corresponding to an optimum cooling water flow rate can be used for subsequent engineering-economic evaluation of the best cooling system variant. The computerized mathematical model and algorithms make it possible to determine the availability and degree of structural changes for the cooling system in all stages of the life cycle of a plant.

  3. Study of numerical schemes for two-phase flow in porous media for any meshes: application to storage of nuclear waste

    International Nuclear Information System (INIS)

    Angelini, O.

    2010-01-01

    The two-phase flow in porous media is a complex phenomenon and which relate to many industrial problems. EDF works on the feasibility and the safety of a storage in deep geologic layer of nuclear waste. In this domain the simulation of the two-phase flow in porous media is particularly important in at least three domains: first of all during the phase of ventilation of the galleries of the storage which could de-saturate the rock and so modify its properties, but also during the phase of re-saturation of the materials and finally during the arrival of the water on the metal parts contained in the storage which will then involve phenomena of corrosion and a hydrogen release. In this context, EDF wishes to obtain robust numerical methods without restrictive condition on the mesh. This work is dedicated at first to the development of the finite volume scheme SUSHI (Scheme Using Stabilization and Hybrid Interfaces) in the code of mechanics of EDF, Code Aster in order to simulate the two-phase flow in porous media. This scheme was developed in 2D and in 3D. At the same time a new formulation which allows to simulate in a uniform way the flows in saturated and unsaturated porous media for miscible and immiscible problems is proposed. Various studies simulating difficulties related to the problems of the storage of nuclear waste in deep geological layers were study. We can quote the study of a bi-material which advances the capillary re-balancing of a material by an other one possessing properties and initial very heterogeneous conditions in saturation. We will also quote the study of the injection of hydrogen in an porous media initially saturated in pure water which is proposed by the benchmark 'two-phase Flow' proposed by the GNR MOMAS. This study had for objective to bring to light the good treatment of the appearance of a phase in a saturated porous media and thus the relevance of our new formulation to study with a way unified a problem of saturated flow and a

  4. Ultrasonic meters in the feedwater flow to recover thermal power in the reactor of nuclear power plant of Laguna Verde U1 and U2

    International Nuclear Information System (INIS)

    Tijerina S, F.

    2008-01-01

    The engineers in nuclear power plants BWRs and PWRs based on the development of the ultrasonic technology for the measurement of the mass, volumetric flow, density and temperature in fluids, have applied this technology in two primary targets approved by the NRC: the use for the recovery of thermal power in the reactor and/or to be able to realize an increase of thermal power licensed in a 2% (MUR) by 1OCFR50 Appendix K. The present article mentions the current problem in the measurement of the feedwater flow with Venturi meters, which affects that the thermal balance of reactor BWRs or PWRs this underestimated. One in broad strokes describes the application of the ultrasonic technology for the ultrasonic measurement in the flow of the feedwater system of the reactor and power to recover thermal power of the reactor. One is to the methodology developed in CFE for a calibration of the temperature transmitters of RTD's and the methodology for a calibration of the venturi flow transmitters using ultrasonic measurement. Are show the measurements in the feedwater of reactor of the temperature with RTD's and ultrasonic measurement, as well as the flow with the venturi and the ultrasonic measurement operating the reactor to the 100% of nominal thermal power, before and after the calibration of the temperature transmitters and flow. Finally, is a plan to be able to realize a recovery of thermal power of the reactor, showing as carrying out their estimations. As a result of the application of ultrasonic technology in the feedwater of reactor BWR-5 in Laguna Verde, in the Unit 1 cycle 13 it was recover an equivalent energy to a thermal power of 25 MWt in the reactor and an exit electrical power of 6 M We in the turbogenerator. Also in the Unit 2 cycle 10 it was recover an equivalent energy to a thermal power of 40 MWt in the reactor and an exit electrical power of 16 M We in the turbogenerator. (Author)

  5. The utility of flow sorting to identify chromosomes carrying a single copy transgene in wheat

    Czech Academy of Sciences Publication Activity Database

    Cápal, Petr; Endo, Takashi R.; Vrána, Jan; Kubaláková, Marie; Karafiátová, Miroslava; Komínková, Eva; Mora-Ramirez, I.; Weschke, W.; Doležel, Jaroslav

    2016-01-01

    Roč. 12, APR 25 (2016), s. 24 ISSN 1746-4811 R&D Projects: GA MŠk(CZ) LO1204; GA ČR GBP501/12/G090 Institutional support: RVO:61389030 Keywords : Transgene localization * Flow cytometric sorting * Single chromosome amplification Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.510, year: 2016

  6. Genome-size variation in switchgrass (Panicum virgatum): flow cytometry and cytology reveal rampant aneuploidy

    Science.gov (United States)

    Switchgrass (Panicum virgatum L.), a native perennial dominant of the prairies of North America, has been targeted as a model herbaceous species for biofeedstock development. A flow-cytometric survey of a core set of 11 primarily upland polyploid switchgrass accessions indicated that there was con...

  7. Predicting corrosion product transport in nuclear power stations using a solubility-based model for flow-accelerated corrosion

    International Nuclear Information System (INIS)

    Burrill, K.A.; Cheluget, E.L.

    1995-01-01

    A general model of solubility-driven flow-accelerated corrosion of carbon steel was derived based on the assumption that the solubilities of ferric oxyhydroxide and magnetite control the rate of film dissolution. This process involves the dissolution of an oxide film due to fast-flowing coolant unsaturated in iron. The soluble iron is produced by (i) the corrosion of base metal under a porous oxide film and (ii) the dissolution of the oxide film at the fluid-oxide film interface. The iron released at the pipe wall is transferred into the bulk flow by turbulent mass transfer. The model is suitable for calculating concentrations of dissolved iron in feedtrain lines. These iron levels were used to calculate sludge transport rates around the feedtrain. The model was used to predict sludge transport rates due to flow accelerated corrosion of major feedtrain piping in a CANDU reactor. The predictions of the model compare well with plant measurements

  8. The Effect of Flow Swirling on the Safety and Reliability of Nuclear Power Installations of New Generation

    Science.gov (United States)

    Mitrofanova, O. V.; Ivlev, O. A.; Urtenov, D. S.

    2018-03-01

    Hydrodynamics and heat exchange in the elements of thermal hydraulic tracts of ship nuclear reactors of the new generation were numerically simulated in this work. Parts of the coolant circuit in the collector and piping systems with geometries that may lead to generation of stable large-scale vortexes, causing a wide range of acoustic oscillations of the coolant, were selected as modeling objects. The purpose of the research is to develop principles of physical and mathematical modeling for scientific substantiation of optimal layout solutions that ensure enhanced operational life of icebreaker’s nuclear power installations of new generation with reactors of integral type.

  9. Groundwater flow modeling of periods with temperate climate conditions for use in a safety assessment of a repository for spent nuclear fuel - 59154

    International Nuclear Information System (INIS)

    Joyce, Steven; Hartley, Lee; Simpson, Trevor

    2012-01-01

    Document available in abstract form only. Full text of publication follows: As a part of the license application for a final repository for spent nuclear fuel, the Swedish Nuclear Fuel and Waste Management Company (SKB) has prepared a safety report (SR-Site) that assesses the long-term radiological safety after closure of a repository located at 500 m depth in the Forsmark area, c. 120 km north of Stockholm. The movement and composition of groundwater affect both the key pathways for radionuclide migration and the performance of engineered barriers, and hence are important issues that have to be considered and modelled as part of quantitative assessment calculations. This presentation describes the groundwater flow modelling studies that have been performed to represent the post-closure hydrogeological and hydrochemical situations during temperate climate conditions, and how these are used to support safety assessment calculations and arguments. The collation and implementation of onsite hydrogeological and hydrogeochemical data from the surface based site investigations at Forsmark are used as the basis for defining a reference case for the natural hydrogeological situation at the site (hydrogeological base case). Areas of uncertainty within the current site understanding and the engineered system are examined by a series of flow model variants

  10. A Novel Multi-Scale Domain Overlapping CFD/STH Coupling Methodology for Multi-Dimensional Flows Relevant to Nuclear Applications

    Science.gov (United States)

    Grunloh, Timothy P.

    The objective of this dissertation is to develop a 3-D domain-overlapping coupling method that leverages the superior flow field resolution of the Computational Fluid Dynamics (CFD) code STAR-CCM+ and the fast execution of the System Thermal Hydraulic (STH) code TRACE to efficiently and accurately model thermal hydraulic transport properties in nuclear power plants under complex conditions of regulatory and economic importance. The primary contribution is the novel Stabilized Inertial Domain Overlapping (SIDO) coupling method, which allows for on-the-fly correction of TRACE solutions for local pressures and velocity profiles inside multi-dimensional regions based on the results of the CFD simulation. The method is found to outperform the more frequently-used domain decomposition coupling methods. An STH code such as TRACE is designed to simulate large, diverse component networks, requiring simplifications to the fluid flow equations for reasonable execution times. Empirical correlations are therefore required for many sub-grid processes. The coarse grids used by TRACE diminish sensitivity to small scale geometric details such as Reactor Pressure Vessel (RPV) internals. A CFD code such as STAR-CCM+ uses much finer computational meshes that are sensitive to the geometric details of reactor internals. In turbulent flows, it is infeasible to fully resolve the flow solution, but the correlations used to model turbulence are at a low level. The CFD code can therefore resolve smaller scale flow processes. The development of a 3-D coupling method was carried out with the intention of improving predictive capabilities of transport properties in the downcomer and lower plenum regions of an RPV in reactor safety calculations. These regions are responsible for the multi-dimensional mixing effects that determine the distribution at the core inlet of quantities with reactivity implications, such as fluid temperature and dissolved neutron absorber concentration.

  11. Diagnosis of regional cerebral blood flow abnormalities using SPECT: agreement between individualized statistical parametric maps and visual inspection by nuclear medicine physicians with different levels of expertise in nuclear neurology

    Energy Technology Data Exchange (ETDEWEB)

    Rocha, Euclides Timoteo da, E-mail: euclidestimoteo@uol.com.b [Fundacao Pio XII, Barretos, SP (Brazil). Hospital de Cancer. Dept. de Medicina Nuclear; Buchpiguel, Carlos Alberto [Hospital do Coracao, Sao Paulo, SP (Brazil). Dept. de Medicina Nuclear; Nitrini, Ricardo [Universidade de Sao Paulo (USP), SP (Brazil). Faculdade de Medicina. Dept. de Neurologia; Tazima, Sergio [Hospital Alemao Oswaldo Cruz (HAOC), Sao Paulo, SP (Brazil). Dept. de Medicina Nuclear; Peres, Stela Verzinhase [Fundacao Pio XII, Barretos, SP (Brazil). Hospital de Cancer; Busatto Filho, Geraldo [Universidade de Sao Paulo (USP), SP (Brazil). Faculdade de Medicina. Div. de Medicina Nuclear

    2009-07-01

    Introduction: visual analysis is widely used to interpret regional cerebral blood flow (rCBF) SPECT images in clinical practice despite its limitations. Automated methods are employed to investigate between-group rCBF differences in research studies but have rarely been explored in individual analyses. Objectives: to compare visual inspection by nuclear physicians with the automated statistical parametric mapping program using a SPECT dataset of patients with neurological disorders and normal control images. Methods: using statistical parametric mapping, 14 SPECT images from patients with various neurological disorders were compared individually with a databank of 32 normal images using a statistical threshold of p<0.05 (corrected for multiple comparisons at the level of individual voxels or clusters). Statistical parametric mapping results were compared with visual analyses by a nuclear physician highly experienced in neurology (A) as well as a nuclear physician with a general background of experience (B) who independently classified images as normal or altered, and determined the location of changes and the severity. Results: of the 32 images of the normal databank, 4 generated maps showing rCBF abnormalities (p<0.05, corrected). Among the 14 images from patients with neurological disorders, 13 showed rCBF alterations. Statistical parametric mapping and physician A completely agreed on 84.37% and 64.28% of cases from the normal databank and neurological disorders, respectively. The agreement between statistical parametric mapping and ratings of physician B were lower (71.18% and 35.71%, respectively). Conclusion: statistical parametric mapping replicated the findings described by the more experienced nuclear physician. This finding suggests that automated methods for individually analyzing rCBF SPECT images may be a valuable resource to complement visual inspection in clinical practice. (author)

  12. Flow cytometric assessment of activation of peripheral blood platelets in dogs with normal platelet count and asymptomatic thrombocytopenia.

    Science.gov (United States)

    Żmigrodzka, M; Guzera, M; Winnicka, A

    2016-01-01

    Platelets play a crucial role in hemostasis. Their activation has not yet been evaluated in healthy dogs with a normal and low platelet count. The aim of this study was to determine the influence of activators on platelet activation in dogs with a normal platelet count and asymptomatic thrombocytopenia. 72 clinically healthy dogs were enrolled. Patients were allocated into three groups. Group 1 consisted of 30 dogs with a normal platelet count, group 2 included 22 dogs with a platelet count between 100 and 200×109/l and group 3 consisted of 20 dogs with a platelet count lower than 100×109/l. Platelet rich-plasma (PRP) was obtained from peripheral blood samples using tripotassium ethylenediaminetetraacetic acid (K3-EDTA) as anticoagulant. Next, platelets were stimulated using phorbol-12-myristate-13-acetate or thrombin, stabilized using procaine or left unstimulated. The expression of CD51 and CD41/CD61 was evaluated. Co-expression of CD41/CD61 and Annexin V served as a marker of platelet activation. The expression of CD41/CD61 and CD51 did not differ between the 3 groups. Thrombin-stimulated platelets had a significantly higher activity in dogs with a normal platelet count than in dogs with asymptomatic thrombocytopenia. Procaine inhibited platelet activity in all groups. In conclusion, activation of platelets of healthy dogs in vitro varied depending on the platelet count and platelet activator.

  13. Development of a flow cytometric immunoassay for recombinant bovine somatotropin-induced antibodies in serum of dairy cows

    NARCIS (Netherlands)

    Smits, N.G.E.; Bremer, M.G.E.G.; Ludwig, S.K.J.; Nielen, M.W.F.

    2012-01-01

    Administration of recombinant bovine somatotropin (rbST) to enhance milk production in dairy cows is banned within the European Union. Therefore, methods for pinpointing rbST abuse are required. Due to the problematic detection of rbST itself in serum, methods are also focused on detecting changes

  14. Transmission Electron Microscopic Morphological Study and Flow Cytometric Viability Assessment of Acinetobacter baumannii Susceptible to Musca domestica cecropin

    OpenAIRE

    Gui, Shuiqing; Li, Rongjiang; Feng, Yongwen; Wang, Sanming

    2014-01-01

    Multidrug-resistant (MDR) Acinetobacter baumannii infections are difficult to treat owing to the extremely limited armamentarium. Expectations about antimicrobial peptides' use as new powerful antibacterial agents have been raised on the basis of their unique mechanism of action. Musca domestica cecropin (Mdc), a novel antimicrobial peptide from the larvae of Housefly (Musca domestica), has potently active against Gram-positive and Gram-negative bacteria standard strain. Here we evaluated the...

  15. Studies of the ABO and FORS Histo-Blood Group Systems: Focus on Flow Cytometric and Genetic Analysis

    OpenAIRE

    Hult, Annika

    2013-01-01

    ABO is the clinically most important blood group system and its antigens are carbohydrate moieties present on the surface of the red blood cell (RBC) but also on other tissues throughout the body. The ABO gene encodes an enzyme, a glycosyltransferase (GT),that adds a terminal monosaccharide to the precursor structure, H antigen, to define the A or B antigens. Blood group O is due to a non-functional GT that leaves the precursor unchanged. Weak expression of ABO antigens can be acquired or be ...

  16. Flow cytometric chromosome sorting from diploid progenitors of bread wheat, T. urartu, Ae. speltoides and Ae. tauschii

    Czech Academy of Sciences Publication Activity Database

    Molnár, I.; Kubaláková, Marie; Šimková, Hana; Farkas, A.; Cseh, A.; Megyeri, M.; Vrána, Jan; Molnár-Láng, M.; Doležel, Jaroslav

    2014-01-01

    Roč. 127, č. 5 (2014), s. 1091-1104 ISSN 0040-5752 R&D Projects: GA ČR GBP501/12/G090; GA MŠk(CZ) LO1204 Grant - others:GA MŠk(CZ) ED0007/01/01 Program:ED Institutional support: RVO:61389030 Keywords : SYNTHETIC HEXAPLOID WHEAT * AEGILOPS-TRITICUM GROUP * GENETIC-LINKAGE MAP Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.790, year: 2014

  17. Single and multigland disease in primary hyperparathyroidism: Clinical follow-up, histopathology, and flow cytometric DNA analysis

    NARCIS (Netherlands)

    H.J. Bonjer (Jaap); H.A. Bruining (Hajo); J.C. Birkenhäger (Jan); Y. Nishiyama (Yoshihiko); M.A. Jones (Michael); C. Bruce Bagwell (C.)

    1992-01-01

    textabstractTwo-hundred seventy-four patients with primary hyperparathyroidism had selective removal of enlarged parathyroid glands. Biopsies were taken from all parathyroid glands. Normal-size glands were not resected irrespective of their histological appearance. After a mean follow-up of 13.5

  18. A rapid flow cytometric method for determining the cellular composition of bronchoalveolar lavage fluid cells in mouse models of asthma

    NARCIS (Netherlands)

    van Rijt, Leonie S.; Kuipers, Harmjan; Vos, Nanda; Hijdra, Daniëlle; Hoogsteden, Henk C.; Lambrecht, Bart N.

    2004-01-01

    Mouse models of allergic asthma are increasingly used to study the immunopathology of this complex disorder. The degree and type of airway inflammation is often studied by determination of differential cell counts on cytospins of bronchoalveolar lavage fluid (BALF) cells stained with May-Grünwald

  19. Immuno-flow cytometric detection of the ichthyotoxic dinoflagellates Gyrodinium aureolum and Gymnodinium nagasakiense : Independence of physiological state

    NARCIS (Netherlands)

    Vrieling, EG; vandePoll, WH; Vriezekolk, G; Gieskes, WWC

    The ichthyotoxic dinoflagellates Gyrodinium aureolum and Gymnodinium nagasakiense were cultured under different environmental conditions to test possible variability in immunochemical labelling intensity of cell-surface antigens using species-specific monoclonal antibodies. Variation of antigen

  20. A modified method of flow cytometric seed screen simplifies the quantification of progeny classes with different ploidy levels

    Czech Academy of Sciences Publication Activity Database

    Krahulcová, Anna; Suda, Jan

    2006-01-01

    Roč. 50, č. 3 (2006), s. 457-460 ISSN 0006-3134 R&D Projects: GA AV ČR IAA6005203 Institutional research plan: CEZ:AV0Z60050516 Keywords : facultative apomixis * reproduction routes * polyhaploids Subject RIV: EF - Botanics Impact factor: 1.198, year: 2006

  1. Single-laboratory validation of a multiplex flow cytometric immunoassay for the simultaneous detection of coccidiostats in eggs and feed

    NARCIS (Netherlands)

    Bienenmann-Ploum, M.E.; Vincent, U.; Campbell, K.; Huet, A.C.; Haasnoot, W.; Delahaut, P.; Stolker, A.A.M.; Elliott, C.T.; Nielen, M.W.F.

    2013-01-01

    Coccidiostats are authorized in the European Union (EU) to be used as poultry feed additives. Maximum (residue) levels (M(R)Ls) have been set within the EU for consumer and animal protection against unintended carry-over, and monitoring is compulsory. This paper describes the single-laboratory

  2. Naturalized plants have smaller genomes than their non-invading relatives: a flow cytometric analysis of the Czech alien flora

    Czech Academy of Sciences Publication Activity Database

    Kubešová, M.; Moravcová, Lenka; Suda, Jan; Jarošík, V.; Pyšek, Petr

    2010-01-01

    Roč. 82, č. 1 (2010), s. 81-96 ISSN 0032-7786 R&D Projects: GA ČR GA206/09/0563; GA ČR GD206/08/H049; GA MŠk LC06073 Institutional research plan: CEZ:AV0Z60050516 Keywords : cytometry * ploidy * genome size Subject RIV: EF - Botanics Impact factor: 2.792, year: 2010

  3. Update report on fracture flow in saturated tuff: Dynamic transport task for the Nevada Nuclear Waste Investigations

    International Nuclear Information System (INIS)

    Janecky, D.R.; Rundberg, R.S.; Ott, M.; Mitchell, A.

    1990-11-01

    This report summarizes the results of continuing experiments on the behavior of tracers during fracture flow in saturated, welded tuff. These experiments were completed during the past year as part of the Dynamic Transport Task of geochemical investigations for the Yucca Mountain Project sponsored by the US Department of Energy. These experiments are designed to investigate the effects of fluid movement in fractures when coupled with matrix diffusion and sorption but isolated from the effects of capillary suction and two-phase flow characteristic of unsaturated conditions. The experiments reported here are continuations of experimental efforts reported previously. The behavior of three tracers [HTO (tritiated water), TcO 4 - (pertechnetate), and sulforhodamine B dye] have been investigated during flow through a saturated column of densely welded tuff from the Topopah Spring Member of the Paintbrush Tuff, Yucca Mountain, Nye County, southern Nevada. 31 refs., 26 figs., 2 tabs

  4. A system for the discharge of gas bubbles from the coolant flow of a nuclear reactor cooled by forced circulation

    International Nuclear Information System (INIS)

    Markfort, D.; Kaiser, A.; Dohmen, A.

    1975-01-01

    In a reactor cooled by forced circulation the gas bubbles carried along with the coolant flow are separated before entering the reactor core or forced away into the external zones. For this purpose the coolant is radially guided into a plenum below the core and deflected to a tangential direction by means of flow guide elements. The flow runs spirally downwards. On the bubbles, during their dwell time in this channel, the buoyant force and a force towards the axis of symmetry of the tank are exerted. The major part of the coolant is directed into a radial direction by means of a guiding apparatus in the lower section of the channel and guided through a chimney in the plenum to the center of the reactor core. This inner chimney is enclosed by an outer chimney for the core edge zones through which coolant with a small share of bubbles is taken away. (RW) [de

  5. Modeling of strongly heat-driven flow processes at a potential high-level nuclear waste repository at Yucca Mountain, Nevada

    International Nuclear Information System (INIS)

    Pruess, K.; Tsang, Y.

    1993-01-01

    Two complementary numerical models for analyzing high-level nuclear waste emplacement at Yucca Mountain have been developed. A vertical cross-sectional (X-Z) model permits a realistic representation of hydrogeologic features, such as alternating tilting layers of welded and non-welded tuffs. fault zones, and surface topography. An alternative radially symmetric (R-Z) model is more limited in its ability to describe the hydrogeology of the site, but is better suited to model heat transfer in the host rock. Our models include a comprehensive description of multiphase fluid and heat flow processes, including strong enhancements of vapor diffusion from pore-level phase change effects. The neighborhood of the repository is found to partially dry out from the waste heat. A condensation halo of large liquid saturation forms around the drying zone, from which liquid flows downward at large rates. System response to infiltration from the surface and to ventilation of mined openings is evaluated. The impact of the various flow processes on the waste isolation capabilities of the site is discussed

  6. Modeling of strongly heat-driven flow processes at a potential high-level nuclear waste repository at Yucca Mountain, Nevada

    International Nuclear Information System (INIS)

    Pruess, K.; Tsang, Y.

    1993-01-01

    Two complementary numerical models for analyzing high-level nuclear waste emplacement at Yucca Mountain have been developed. A vertical cross-sectional (X-Z) model permits a realistic representation of hydrogeologic features, such as alternating tilting layers of welded and non-welded tuffs, fault zones, and surface topography. An alternative radially symmetric (R-Z) model is more limited in its ability to describe the hydrogeology of the site, but is better suited to model heat transfer in the host rock. Our models include a comprehensive description of multiphase fluid and heat flow processes, including strong enhancements of vapor diffusion from pore-level phase change effects. The neighborhood of the repository is found to partially dry out from the waste heat. A condensation halo of large liquid saturation forms around the drying zone, from which liquid flows downward at large rates. System response to infiltration from the surface and to ventilation of mined openings is evaluated. The impact of the various flow processes on the waste isolation capabilities of the site is discussed

  7. A semi-analytical study of the vibrations induced by flow in the piping of nuclear power plants

    International Nuclear Information System (INIS)

    Maneschy, J.E.

    1981-01-01

    A semi-analytical method is presented to evaluate the piping system safety due to internal flow vibration excitation. The method is based on the application of a plane spectrum on the system, resulted by measured modal accelerations. A criteria is established to verify stress levels and compare with the allowable levels. (Author) [pt

  8. Modeling analyses of two-phase flow instabilities for straight and helical tubes in nuclear power plants

    International Nuclear Information System (INIS)

    Dong, Ruiting; Niu, Fenglei; Zhou, Yuan; Yu, Yu; Guo, Zhangpeng

    2016-01-01

    Highlights: • Two-phase flow instabilities in straight and helical tubes were studied. • The effects of system pressure, mass flux, inlet subcooling on DWO were studied. • The simulation results are consistent with the experimental results. • The RELAP5 results are consistent with frequency domain method results. - Abstract: The effects of system pressure, mass flux and inlet subcooling on two-phase flow instability for the test section consisted of two heated straight channels or two helical channels are studied by means of RELAP5/MOD3.3 and multi-variable frequency domain control theory. The experimental data in two straight channels are used to verify the RELAP5 and multi-variable frequency domain control theory results. The thermal hydraulic behaviors and parametric effects are simulated and compared with the experimental data. The RELAP5 results show that the flow stability increases with the system pressure, mass velocity, and inlet subcooling at high subcoolings. The frequency domain theory presents the same results as those given by the time domain theory (RELAP5). The effects of system pressure, mass velocity and inlet subcooling are simulated to find the difference between the straight and the helical tube flows. The RELAP5 and the multi-variable frequency domain control theory are used in modeling and simulating density wave oscillation to study their advantages and disadvantages in straight and helical tubes.

  9. Restorative proctocolectomy: histological assessment and cytometric DNA analysis of ileal pouch biopsies.

    Science.gov (United States)

    Pronio, A; Montesani, C; Vecchione, A; Giovagnoli, M G; Giarnieri, E; Nardi, F; Nigri, G; Ribotta, G

    1997-01-01

    The pathological changes and the risk of developing cancer in the ileal pouch mucosa of patients who received restorative proctocolectomy with ileal pouch-anal anastomosis (IPAA) were studied. The presence or absence of remaining rectal mucosa below the IPAA in both patients with stapled and handsewn IPAA was also examined. Endoscopy of the ileal pouch was performed on 38 patients at 4, 12, 18 and 36 months after restorative proctocolectomy with ileal pouch. Mucosal biopsy specimens were taken from the ileal reservoir in order to assess the histological incidence of inflammation. In 23 patients, biopsies were taken to perform cytometric DNA analysis. Clinical symptoms of pouchitis (over six evacuations in 24 hours, night-time evacuations, leakage of feces, bloody diarrhea, abdominal pain and fever) were recorded and correlated with the histological findings. Biopsies were also sampled below the ileo-anal anastomosis (IPAA) in order to identify residual rectal mucosa. Results of histological assessment showed various degrees of chronic inflammation increasing over time (from 42 to 60%) while the presence of both acute and chronic inflammation of the reservoir was less frequent (from 18 to 30%). Villous atrophy was present in 39-68% of patients and the grade of villous atrophy was correlated to the grade of inflammation. Clinical pouchitis was present in 3 to 8% of cases at the different controls and it was always associated with the highest grade of histological inflammation and severe villous atrophy. No significant alteration of the DNA cellular content was observed. Very low incidence of aneuploidy (0.7-1% Ex.R.) has been reported in three cases. However, we found dysplasia in only one patient who underwent surgical treatment for familial polyposis coli. IPAA evaluation showed no residual rectal mucosa in 40% of cases with stapled IPAA; in the remaining 60%, we found a small amount of rectal mucosa (maximum 1 cm). We did not find rectal mucosa after handsewn IPAA

  10. MODELING OF FLOW AND TRANSPORT INDUCED BY PRODUCTION OF HYDROFRACTURE-STIMULATED GAS WELLS NEAR THE RULISON NUCLEAR TEST

    Energy Technology Data Exchange (ETDEWEB)

    Hodges, Rex A. [Navarro Research and Engineering; Cooper, Clay [Desert Research Inst. (DRI), Reno, NV (United States); Falta, Ronald [Clemson Univ., SC (United States)

    2012-09-17

    The Piceance Basin in western Colorado contains significant reserves of natural gas in poorly connected, low-permeability (tight) sandstone lenses of the Mesaverde Group. The ability to enhance the production of natural gas in this area has long been a goal of the oil and gas industry. The U.S. Atomic Energy Commission, a predecessor agency to the U.S. Department of Energy (DOE) and the U.S. Nuclear Regulatory Commission, participated in three tests using nuclear detonations to fracture tight formations in an effort to enhance gas production. The tests were conducted under Project Plowshare, a program designed to identify peaceful, beneficial uses for nuclear devices. The first, Project Gasbuggy, was conducted in 1967 in the San Juan Basin of New Mexico. The two subsequent tests, Project Rulison in 1969 and Project Rio Blanco in 1973, were in the Piceance Basin. The ability to enhance natural gas production from tight sands has become practical through advances in hydraulic fracturing technology (hydrofracturing). This technology has led to an increase in drilling activity near the Rulison site, raising concerns that contamination currently contained in the subsurface could be released through a gas well drilled too close to the site. As wells are drilled nearer the site, the DOE Office of Legacy Management has taken the approach outlined in the June 2010 Rulison Path Forward document (DOE 2010), which recommends a conservative, staged approach to gas development. Drillers are encouraged to drill wells in areas with a low likelihood of encountering contamination (both distance and direction from the detonation zone are factors) and to collect data from these wells prior to drilling nearer the site’s 40 acre institutional control boundary (Lot 11). Previous modeling results indicate that contamination has been contained within Lot 11 (Figure 1). The Path Forward document couples the model predictions with the monitoring of gas and produced water from the gas wells

  11. Supercontinuum white light lasers for flow cytometry

    Science.gov (United States)

    Telford, William G.; Subach, Fedor V.; Verkhusha, Vladislav V.

    2009-01-01

    Excitation of fluorescent probes for flow cytometry has traditionally been limited to a few discrete laser lines, an inherent limitation in our ability to excite the vast array of fluorescent probes available for cellular analysis. In this report, we have used a supercontinuum (SC) white light laser as an excitation source for flow cytometry. By selectively filtering the wavelength of interest, almost any laser wavelength in the visible spectrum can be separated and used for flow cytometric analysis. The white light lasers used in this study were integrated into a commercial flow cytometry platform, and a series of high-transmission bandpass filters used to select wavelength ranges from the blue (~480 nm) to the long red (>700 nm). Cells labeled with a variety of fluorescent probes or expressing fluorescent proteins were then analyzed, in comparison with traditional lasers emitting at wavelengths similar to the filtered SC source. Based on a standard sensitivity metric, the white light laser bandwidths produced similar excitation levels to traditional lasers for a wide variety of fluorescent probes and expressible proteins. Sensitivity assessment using fluorescent bead arrays confirmed that the SC laser and traditional sources resulted in similar levels of detection sensitivity. Supercontinuum white light laser sources therefore have the potential to remove a significant barrier in flow cytometric analysis, namely the limitation of excitation wavelengths. Almost any visible wavelength range can be made available for excitation, allowing access to virtually any fluorescent probe, and permitting “fine-tuning” of excitation wavelength to particular probes. PMID:19072836

  12. Hybrid approach for fault diagnosis based on multilevel flow model and information fusion of nuclear power plant

    International Nuclear Information System (INIS)

    Ma Jie; Guo Lifeng; Zhang Yusheng; Peng Qiao; Ruan Minzhi

    2011-01-01

    In order to improve the ability of condition monitoring and fault diagnostic system, a hybrid intelligent diagnostic system based on multilevel flow model (MFM) and information fusion was proposed. This method utilized information fusion technique to improve the rapidness and veracity of fault diagnosis, and made use of MFM to explain the alarm propagation path, which could enhance the comprehension of diagnostic result. The emulation test proves that the hybrid intelligent diagnostic system can identify fault and propose the alarm analysis quickly. (authors)

  13. Development and Validation of Methodology to Model Flow in Ventilation Systems Commonly Found in Nuclear Facilities - Phase II

    Energy Technology Data Exchange (ETDEWEB)

    Strons, Philip [Argonne National Lab. (ANL), Argonne, IL (United States); Bailey, James L. [Argonne National Lab. (ANL), Argonne, IL (United States); Davis, John [Argonne National Lab. (ANL), Argonne, IL (United States); Grudzinski, James [Argonne National Lab. (ANL), Argonne, IL (United States); Hlotke, John [Argonne National Lab. (ANL), Argonne, IL (United States)

    2016-03-01

    In this report we present the results of the Phase II analysis and testing of the flow patterns encountered in the Alpha Gamma Hot Cell Facility (AGHCF), as well as the results from an opportunity to expand upon field test work from Phase I by the use of a Class IIIb laser. The addition to the Phase I work is covered before proceeding to the results of the Phase II work, followed by a summary of findings.

  14. Nuclear instrumentation

    International Nuclear Information System (INIS)

    Weill, Jacky; Fabre, Rene.

    1981-01-01

    This article sums up the Research and Development effort at present being carried out in the five following fields of applications: Health physics and Radioprospection, Control of nuclear reactors, Plant control (preparation and reprocessing of the fuel, testing of nuclear substances, etc.), Research laboratory instrumentation, Detectors. It also sets the place of French industrial activities by means of an estimate of the French market, production and flow of trading with other countries [fr

  15. Two-fluid model of two-phase flow in a pin bundle of a nuclear reactor

    International Nuclear Information System (INIS)

    Chawla, T.C.; Ishii, M.

    1980-01-01

    By considering two-phase flow as a field which is subdivided into two turbulent single-phase regions with moving boundaries separating the two constituent phases, such that the differential balances for three-dimensional turbulent flow hold for each subregion and for the interface, we perform the Eulerian area averaging over the cross-sectional area of each phase in a given channel and segment averaging of transverse momentum equation along the phase intercepts at the interchannel boundaries. To simplify the governing equations obtained as a result of these operations, we invoke the assumption that the motion of the fluid in each phase is dominantly in axial direction, that is the transverse components of velocity are small compared to axial components. We further assume that the variation of axial component of velocity within a channel is much stronger than the variation along the axial direction. We also assume that similar arguments can also be applied to the variation of enthalpy in a channel. As a result of these considerations, we obtain two sets of continuity, momentum, and energy equations describing motion of each phase in the axial direction. The phasic interaction terms which appear in these equations are governed by interfacial transfer conditions obtained from interface balances. The segment-averaged transverse-momentum equation for each phase provides the governing equation for cross flow. (author)

  16. Application of qualitative reasoning with functional knowledge represented by Multilevel Flow Modeling to diagnosis of accidental situation in nuclear power plant

    International Nuclear Information System (INIS)

    Yoshida, Kazuo; Tanabe, Fumiya; Kawase, Katumi.

    1996-01-01

    It has been proposed to use the Multilevel Flow Modeling (MFM) by M. Lind as a framework for functional knowledge representation for qualitative reasoning in a complex process system such as nuclear power plant. To build a knowledge base with MFM framework makes it possible to represent functional characteristics in different levels of abstraction and aggregation. A pilot inference system based on the qualitative reasoning with MFM has been developed to diagnose a cause of abnormal events in a typical PWR power plant. Some single failure events has been diagnosed with this system to verify the proposed method. In the verification study, some investigation has been also performed to clarify the effects of this knowledge representation in efficiency of reasoning and ambiguity of qualitative reasoning. (author)

  17. Study of the Flow Temperature and Ring Design Influence on the Response of a New Reduced-Size Calorimetric Cell for Nuclear Heating Quantification

    Science.gov (United States)

    Volte, A.; Reynard-Carette, C.; Brun, J.; De Vita, C.; Carette, M.; Fiorido, T.; Lyoussi, A.; Fourmentel, D.; Villard, J.-F.; Guimbal, P.

    2018-01-01

    This paper concerns experimental studies of different designs of a new compact calorimetric cell under laboratory conditions. This kind of cell is used for the measurement of the nuclear heating rate inside Material Testing Reactors thanks to differential calorimetry. The results, obtained by applying an operating protocol corresponding to a preliminary out-of-pile calibration step, are presented for three designs. The influence of the horizontal-fin design is shown on the calibration curve and the sensor sensitivity. The influence of the external fluid flow temperature is given for the quarter design. The different responses of the calorimetric cell are explained by taken into account a 1D analytical thermal model coupling thermal conductive and radiative transfers.

  18. Method and means for abruptly terminating the flow of fluid in closed circulating systems of nuclear reactor plants or the like

    International Nuclear Information System (INIS)

    Schiele, O.; Florjancic, D.

    1976-01-01

    A nuclear steam supply system is described wherein each of a plurality of centrifugal pumps begins to operate with full cavitation in response to an abrupt drop of system pressure in the event of leakage. This is achieved by influencing the net positive suction head of each pump over the entire range of fluid flow and/or by influencing the net positive suction head upstream of the pumps. The first mode of causing the pumps to operate with full cavitation includes an appropriate selection of the inlet angle and/or inlet diameter of the pump impeller, the provision of auxiliary impellers which are located upstream of the pumps and can circulate the fluid in or counter to the direction of rotation of the respective pump impellers, or the provision of suitably curved guide vanes in the pumps. The second mode include interrupting the admission of undercooled fluid into the system upstream of the pumps

  19. DIFFERENTIATION OF AURANTII FRUCTUS IMMATURUS AND FRUCTUS PONICIRI TRIFOLIATAE IMMATURUS BY FLOW-INJECTION WITH ULTRAVIOLET SPECTROSCOPIC DETECTION AND PROTON NUCLEAR MAGNETIC RESONANCE USING PARTIAL LEAST-SQUARES DISCRIMINANT ANALYSIS

    OpenAIRE

    Zhang, Mengliang; Zhao, Yang; Harrington, Peter de B.; Chen, Pei

    2015-01-01

    Two simple fingerprinting methods, flow-injection coupled to ultraviolet spectroscopy and proton nuclear magnetic resonance, were used for discriminating between Aurantii fructus immaturus and Fructus poniciri trifoliatae immaturus. Both methods were combined with partial least-squares discriminant analysis. In the flow-injection method, four data representations were evaluated: total ultraviolet absorbance chromatograms, averaged ultraviolet spectra, absorbance at 193, 205, 225, and 283 nm, ...

  20. The management of the flow assisted corrosion on the nuclear power plants of the fleet of EDF: a strategy of commitment for the control of the ageing

    International Nuclear Information System (INIS)

    Trevin, S.; Moutrille, M.P.; Caylar, P.; Delacoux, D.; Gipon, E.; Mellin, N.; Qiu, G.; Alos Ramos, O.; Tarabay, J.; Mansour, C.

    2015-01-01

    Flow accelerated corrosion (FAC) is very effective for nuclear power plants. This generalized corrosion can lead to the rupture of pipes. EDF has developed during these 20 last years the software called BRT-CICERO for the surveillance of the secondary piping system of its Pressurized Water Reactors (PWRs). This software enables the operator to calculate the FAC wear rates taking into account all the influencing parameters such as: pipe isometrics, chromium content of the steel, chemical conditioning and operating parameters of the secondary circuit (temperature, pressure, etc.). This is a major tool for the operators to organize the maintenance and to plan the inspections. In the framework of the French pressure vessel law issued on March 15. 2000, the software BRT-CICERO has been recognized by the French authority for the FAC surveillance on the secondary pressure piping lines of the EDF 58 nuclear reactors. It takes advantage of the experience feedback of the French fleet and of the improvements. BRT-CICERO is frequently updated. A guideline is associated to BRT-CICERO to define the rules and the domain of use and to take advantage of all its functionalities. (authors)

  1. Integration of genetic and physical maps of the chickpea (Cicer arietinum L.) genome using flow-sorted chromosomes

    Czech Academy of Sciences Publication Activity Database

    Zatloukalová, Pavlína; Hřibová, Eva; Kubaláková, Marie; Suchánková, Pavla; Šimková, Hana; Adoración, C.; Kahl, G.; Millán, T.; Doležel, Jaroslav

    2011-01-01

    Roč. 19, č. 6 (2011), s. 729-739 ISSN 0967-3849 R&D Projects: GA MŠk(CZ) LC06004 Institutional research plan: CEZ:AV0Z50380511 Keywords : BAC-FISH * Chromosome isolation * Flow cytometric sorting Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.087, year: 2011

  2. Detection of Xanthomonas axonopodis pv. phaseoli in bean seeds by flow cytometry, immunostaining and direct viable counting

    NARCIS (Netherlands)

    Tebaldi, N.D.; Peters, J.; Chitarra, L.G.; Souza, R.M.; Zouwen, van der P.S.; Bergervoet, J.H.W.; Wolf, van der J.M.

    2010-01-01

    Flow cytometric analysis of immuno-stained cells (immuno-FCM) was compared to immunofluorescence microscopy (IF) and dilution plating on a semi-selective medium, for quantitative detection of Xanthomonas axonopodis pv. phaseoli (Xap) in bean seed extracts. Cell concentrations of Xap between 103-107

  3. Flow cytogenetic studies in chromosomes and whole cells for the detection of clastogenic effects

    International Nuclear Information System (INIS)

    Otto, F.J.; Oldiges, H.

    1980-01-01

    Flow cytometric measurements of the chromosomal DNA content have been used to develop a screening method for the detection of chemically- or physically-induced cytogenetic damage. The reproducibility of this flow cytogenetic assay was shown in a series of subcultures of a Chinese hamster cell clone. The accuracy and sensitivity was tested in cultures treated with chemical mutagens and x-rays. The clastogenic effectiveness was quantified and the dose-effect relationship was established by the increase of the coefficient of variation of the peak of the largest chromosome type in the flow histograms. Since structural chromosome aberrations cause an unequal division of the DNA at mitosis, it is expected that clastogenic effects can be detected also in whole cells of growing populations as an increased dispersion of the cellular DNA content. In order to test this feature, high resolution flow cytometric measurements were performed in x-irradiated hamster cells in vitro and mouse bone marrow cells in vivo

  4. Measuring flow and pressure of lithium coolant under developmental testing of a high-temperature cooling system of a space nuclear power plant

    Science.gov (United States)

    Sobolev, V. Ya.; Sinyavsky, V. V.

    2014-12-01

    Sub-megawatt space NPP use lithium as a coolant and niobium alloy as a structural material. In order to refine the lithium-niobium technology of the material and design engineering, lithium-niobium loops were worked out in RSC Energia, and they were tested at a working temperature of lithium equal to 1070-1300 K. In order to measure the lithium flow and pressure, special gauges were developed, which made possible the calibration and checkout of the loops without their dismantling. The paper describes the architecture of the electromagnetic flowmeter and the electromagnetic vibrating-wire pressure transducer (gauge) for lithium coolant in the nuclear power plant cooling systems. The operating principles of these meters are presented. Flowmeters have been developed for channel diameters ranging from 10 to 100 mm, which are capable of measuring lithium flows in the range of 0.1 to 30 L/s with the error of 3% for design calibration and 1% for volume graduation. The temperature error of the pressure transducers does not exceed 0.4% per 100 K; the nonlinearity and hysteresis of the calibration curve do not exceed 0.3 and 0.4%, respectively. The transducer applications are illustrated by the examples of results obtained from tests on the NPP module mockup and heat pipes of a radiation cooler.

  5. Application of Nano-Structured Coatings for Mitigation of Flow-Accelerated Corrosion in Secondary Pipe Systems of Nuclear Power Plants

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Seung Hyun; Kim, Jong Jin; Yoo, Seung Chang; Huh, Jae Hoon; Kim, Ji Hyun [Ulsan National Institute of Science and Technology, Ulsan (Korea, Republic of)

    2014-05-15

    Flow-accelerated corrosion (FAC) is a complex corrosion process combined with mechanical reaction with fluid. There were lots of research to mitigate FAC such as controlling temperature or water chemistry but in this research, we adopt active coating techniques especially nano-particle reinforced coatings. One of the general characteristics of FAC and its mitigation is that surface friction due to surface morphology makes a significant effect on FAC. Therefore to form a uniform coating layers, nano-particles including TiO2, SiC, Fe-Cr-W and Graphene were utilized. Those materials are known as greatly improve the corrosion resistance of substrates such as carbon steels but their effects on mitigation of FAC are not revealed clearly. Therefore in this research, the FAC resistive performance of nano-structured coatings were tested by electrochemical impedance spectroscopy (EIS) in room temperature 15 wt% sulfuric acid. As the flow-accelerated corrosion inhibitors in secondary piping system of nuclear power plants, various kinds of nano-structured coatings were prepared and tested in room-temperature electrochemical cells. SHS7740 with two types of Densifiers, electroless nickel plating with TiO2 are prepared. Electropolarization curves shows the outstanding corrosion mitigation performance of SHS7740 but EIS results shows the promising potential of Ni-P and Ni-P-TiO2 electroless nickel plating. For future work, high-temperature electrochemical analysis system will be constructed and in secondary water chemistry will be simulated.

  6. Study of the nuclear matter flow with the multidetector INDRA; Etude de l`ecoulement de matiere nucleaire avec le multidetecteur INDRA

    Energy Technology Data Exchange (ETDEWEB)

    Lefort, T. [Caen Univ., 14 (France). Lab. de Physique Corpusculaire

    1997-06-12

    The work presented in this thesis relates to the study of the products which are not emitted by the statistical deexcitation of the projectile and target. The experiment on which this work is based on was performed at GANIL with the detector INDRA on two systems: {sup 36} Ar + {sup 58} Ni and {sup 129} Xe + {sup 119} Sn. A whole characterization (mass, composition, multiplicities and energy properties) of mid-rapidity emission has been done for the system {sup 36} Ar + {sup 58} Ni between 52 and 95 A.MeV. The amount of matter associated to this emission seems to be independent of the incident energy and directly linked with the centrality of the collision. The available energy per nucleon for the production of mid-rapidity products seems to be insensitive to the impact parameter. A systematic study of the nuclear matter in-plane flow has also been carried out. It has been established that the usual methods for reaction plane determination do not allow one to measure accurately the value of flow parameter at intermediate energies. Nevertheless, the inversion energy of the system {sup 36} Ar + {sup 58} Ni can be calculated for central collisions. This energy is independent of the nature of the products. Its value and the features of the mid-rapidity emission should allow one to extract in-medium nucleon-nucleon cross section by comparison with theoretical results. (author) 80 refs.

  7. Numerical and experimental investigations on flow through perforated plates similar to the ones of the bottom end piece of nuclear reactors

    International Nuclear Information System (INIS)

    Barros Filho, Jose A.; Navarro, Moyses A.; Santos, Andre A. Campagnole dos

    2008-01-01

    There is an increasing interest in the application of three dimensional computational fluid dynamics (CFD) codes in the design and safety analysis of nuclear power plants. At present, the main objective with respect to CFD codes is generally to improve confidence in the available analysis tools and to achieve a more reliable approach to safety relevant issues. Aiming towards these goals, we present here the results of the first part of a research program devised to develop and validate a CFD methodology to simulate the water flow through perforated plates with geometry similar to the ones of the bottom end piece of the fuel elements of a Pressurized Water Reactor. This component drives the coolant into the core, promotes an adequate mixing of dissolved substances and holds the fuel assembly mechanically in place. It accounts for a great part of the pressure loss through the bottom end piece. We present here the experimental pressure loss of water flow through perforated plates where geometric features like diameter of holes and inlet chamfers were evaluated. The recovering pressure profile downstream of the plates was also measured. The experimental results were compared with numerical modeling performed with the commercial code CFX 11.0. (author)

  8. Application of nano-structured coatings to mitigate flow-accelerated corrosion in secondary pipe systems of nuclear power plants

    International Nuclear Information System (INIS)

    Kim, Seung Hyun; Kim, Jong Jin; Yoo, Seung Chang; Kim, Ji Hyun

    2014-01-01

    Carbon steel is widely used as a structural material in secondary pipe systems. However, the passivity of carbon steel is not sufficient for protection in secondary water chemistry with a very fast-flowing fluid because of the dissolution of ferrous and magnetite ions and surface friction at the interface of the coolant and pipe surface. There have been many efforts to mitigate flow-accelerated corrosion through adoption of advanced water chemistries such as optimized dissolve oxygen (DO) concentration and temperature, as well as usage of new additives such as monoethanol amine (ETA) to adjust pH. However, these mitigation techniques pose certain challenges relating to the compatibility of new water chemistries with the steam generator, the thermal efficiency of the secondary side, etc. In this study, to improve the passivity of carbon steel, nanostructured coatings especially nanoparticle-enhanced surface coatings were adopted to improve resistance to corrosion and wear. Nanoparticles in the coating matrix help decrease the electrochemical potential compared coatings without nanoparticles, and thus help improve the mechanical properties, especially hardness, through precipitation. In other words, nanoparticle-enhanced surface coatings have the potential to mitigate flow-accelerated corrosion in secondary pipe systems. As candidate coatings, TiO 2 - and SiC-enhanced electrolytic and electroless nickel plating and Fe-Cr-W amorphous metallic coatings (AMC) were selected by acquiring the Pourbaix diagram with thermodynamic calculations. Both TiO 2 and SiC show a stable state in secondary water chemistry, and it is estimated that Fe-Cr-W can be applied to secondary water chemistry because it has a similar chemical composition to carbon steel. Electron microscopic analysis results with scanning electron microscopy (SEM) and tunneling electron microscopy (TEM) show the distribution of TiO 2 nanoparticles in the nickel matrix coating layer, whereas the SiC nanoparticles

  9. Local and integral ulrasonic gauges for two-phase flow instrumentation in nuclear reactor and safety technology. Final report

    International Nuclear Information System (INIS)

    Chochlov, V.N.; Duncev, A.V.; Ivanov, V.V.; Kontelev, V.V.; Melnikov, V.I.; Stoppel, L.K.; Prasser, H.M.; Zippe, W.; Zschau, J.; Zboray, R.

    2000-11-01

    The present project was executed in the frame of a co-operation agreement between FZR and the scientific group of Prof. Melnikov of the technical University of Nishny Novgorod (TUNN) in the Russian Federation. It is part of the Federal Government's programme for the provision of advice for Eastern Europe on the building up of democracy and social market economy (TRANSFORM Programme). New methods of two-phase flow instrumentation were developed: Intrusive wave-guide probes can be used for local void fraction measurements. The new ultrasonic mesh sensors allow a fast two-phase flow visualisation with about 250 frames per second. Experiments carried out at the test loop in Rossendorf, but also the tests at the DESIRE facility in Delft have shown that both local wave-guide probes and ultrasonic mesh sensors can be successfully applied under the conditions of high pressure and temperature steam-water mixture, as well as in organic liquids and refrigerants. Furthermore, non-intrusive wave-guide sensors as well as density sensors based on the measurement of the wave propagation velocity in wave-guides immersed into the measuring liquid were developed and tested. In the present stage of the development, the non-intrusive sensors can rather be used for a qualitative gas respectively level detection than for void fraction measurements. The wave-guide density sensor was successfully demonstrated that it is able to measure densities of single-phase liquids. It requires further development of the electronic circuitry. The main innovation was achieved by the development of the ultrasonic mesh sensor, the resoluting capability of which is comparable to methods like electrical wire-mesh sensors and ultra-fast X-ray tomography, while the device itself is robust and low expensive. (orig.) [de

  10. Model depiction of the atmospheric flows of radioactive cesium emitted from the Fukushima Daiichi Nuclear Power Station accident

    Science.gov (United States)

    Nakajima, Teruyuki; Misawa, Shota; Morino, Yu; Tsuruta, Haruo; Goto, Daisuke; Uchida, Junya; Takemura, Toshihiko; Ohara, Toshimasa; Oura, Yasuji; Ebihara, Mitsuru; Satoh, Masaki

    2017-12-01

    In this study, a new method is proposed for the depiction of the atmospheric transportation of the 137Cs emitted from the Fukushima Daiichi Nuclear Power Station accident. This method employs a combination of the results of two aerosol model ensembles and the hourly observed atmospheric 137Cs concentration at surface level during 14-23 March 2011 at 90 sites in the suspended particulate matter monitoring network. The new method elucidates accurate transport routes and the distribution of the surface-level atmospheric 137Cs relevant to eight plume events that were previously identified. The model ensemble simulates the main features of the observed distribution of surface-level atmospheric 137Cs. However, significant differences were found in some cases, and this suggests the need to improve the modeling of the emission scenario, plume height, wet deposition process, and plume propagation in the Abukuma Mountain region. The contributions of these error sources differ in the early and dissipating phases of each event, depending on the meteorological conditions.

  11. Modeling Groundwater Flow and Transport of Radionuclides at Amchitka Island's Underground Nuclear Tests: Milrow, Long Shot, and Cannikin

    Energy Technology Data Exchange (ETDEWEB)

    Ahmed Hassan; Karl Pohlmann; Jenny Chapman

    2002-11-19

    Since 1963, all United States nuclear tests have been conducted underground. A consequence of this testing has been the deposition of large amounts of radioactive material in the subsurface, sometimes in direct contact with groundwater. The majority of this testing occurred on the Nevada Test Site (NTS), but a limited number of experiments were conducted in other locations. One of these locations, Amchitka Island, Alaska is the subject of this report. Three underground nuclear tests were conducted on Amchitka Island. Long Shot was an 80-kiloton-yield test conducted at a depth of 700 meters (m) on October 29, 1965 (DOE, 2000). Milrow had an announced yield of about 1,000 kilotons, and was detonated at a depth of 1,220 m on October 2, 1969. Cannikin had an announced yield less than 5,000 kilotons, and was conducted at a depth of 1,790 m on November 6, 1971. The purpose of this work is to provide a portion of the information needed to conduct a human-health risk assessment of the potential hazard posed by the three underground nuclear tests on Amchitka Island. Specifically, the focus of this work is the subsurface transport portion, including the release of radionuclides from the underground cavities and their movement through the groundwater system to the point where they seep out of the ocean floor and into the marine environment. This requires a conceptual model of groundwater flow on the island using geologic, hydrologic, and chemical information, a numerical model for groundwater flow, a conceptual model of contaminant release and transport properties from the nuclear test cavities, and a numerical model for contaminant transport. Needed for the risk assessment are estimates of the quantity of radionuclides (in terms of mass flux) from the underground tests on Amchitka that could discharge to the ocean, the time of possible discharge, and the location in terms of distance from shoreline. The radionuclide data presented here are all reported in terms of normalized

  12. Nuclear reactor

    International Nuclear Information System (INIS)

    Mysels, K.J.; Shenoy, A.S.

    1976-01-01

    A nuclear reactor is described in which the core consists of a number of fuel regions through each of which regulated coolant flows. The coolant from neighbouring fuel regions is combined in a manner which results in an averaging of the coolant temperature at the outlet of the core. By this method the presence of hot streaks in the reactor is reduced. (UK)

  13. Modeling and numerical simulation of oscillatory two-phase flows, with application to boiling water nuclear reactors

    Energy Technology Data Exchange (ETDEWEB)

    Rosa, M.P. [Instituto de Estudos Avancados - CTA, Sao Paolo (Brazil); Podowski, M.Z. [Rensselaer Polytechnic Institute, Troy, NY (United States)

    1995-09-01

    This paper is concerned with the analysis of dynamics and stability of boiling channels and systems. The specific objectives are two-fold. One of them is to present the results of a study aimed at analyzing the effects of various modeling concepts and numerical approaches on the transient response and stability of parallel boiling channels. The other objective is to investigate the effect of closed-loop feedback on stability of a boiling water reactor (BWR). Various modeling and computational issues for parallel boiling channels are discussed, such as: the impact of the numerical discretization scheme for the node containing the moving boiling boundary on the convergence and accuracy of computations, and the effects of subcooled boiling and other two-phase flow phenomena on the predictions of marginal stability conditions. Furthermore, the effects are analyzed of local loss coefficients around the recirculation loop of a boiling water reactor on stability of the reactor system. An apparent paradox is explained concerning the impact of changing single-phase losses on loop stability. The calculations have been performed using the DYNOBOSS computer code. The results of DYNOBOSS validation against other computer codes and experimental data are shown.

  14. Feasibility Study on Nano-structured Coatings to Mitigate Flow-accelerated Corrosion in Secondary System of Nuclear Power Plants

    International Nuclear Information System (INIS)

    Kim, Seunghyun; Kim, Jeong Won; Kim, Ji Hyun

    2015-01-01

    There have been many efforts to mitigate FAC through the adoption of the advanced and modified water chemistries such as optimized dissolved oxygen (DO) concentration and temperature. However, these mitigation techniques pose certain challenges relating to the compatibility of new water chemistries with the steam generator, the thermal efficiency of the secondary side, etc. In this context, nano-particle reinforced electroless nickel plating (NP ENP) could help solve the FAC issues in secondary pipe systems. This does not require modification of water chemistry or structural materials, and hence, its application is reasonable and time-saving compared to previous FAC mitigation techniques. The main parameters of FAC are known as electrochemical reaction at the interface, dissolution of magnetite and ferrous ions due to concentration gradient between carbon steels and water and wear due to a fast-flowing fluid. High-temperature corrosion characteristics of the both coatings have potential as FAC barrier for carbon steel. Feasibility study will be carried out with FAC simulation experiments

  15. Feasibility Study on Nano-structured Coatings to Mitigate Flow-accelerated Corrosion in Secondary System of Nuclear Power Plants

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Seunghyun; Kim, Jeong Won; Kim, Ji Hyun [Ulsan National Institute of Science and Technology, Banyeon-ri, Eonyang-eup, Ulju-gun, Ulsan (Korea, Republic of)

    2015-05-15

    There have been many efforts to mitigate FAC through the adoption of the advanced and modified water chemistries such as optimized dissolved oxygen (DO) concentration and temperature. However, these mitigation techniques pose certain challenges relating to the compatibility of new water chemistries with the steam generator, the thermal efficiency of the secondary side, etc. In this context, nano-particle reinforced electroless nickel plating (NP ENP) could help solve the FAC issues in secondary pipe systems. This does not require modification of water chemistry or structural materials, and hence, its application is reasonable and time-saving compared to previous FAC mitigation techniques. The main parameters of FAC are known as electrochemical reaction at the interface, dissolution of magnetite and ferrous ions due to concentration gradient between carbon steels and water and wear due to a fast-flowing fluid. High-temperature corrosion characteristics of the both coatings have potential as FAC barrier for carbon steel. Feasibility study will be carried out with FAC simulation experiments.

  16. Optimal power flow in a power system with nuclear option using multi-objective function: fuel cost and flat voltage profile

    International Nuclear Information System (INIS)

    Rizki Firmansyah Setya Budi; Sarjiya; Sasongko Pramono Hadi

    2016-01-01

    The purpose of power system operation is to supply power with good quality and minimum generation cost. Quality requires cost hence to obtain such purpose, an optimization with objective functions that target on maximizing cost and, at the same time, minimizing cost needs to be carried out. The objective of the research is to obtain optimal power flow (OPF) condition in terms of generation cost and power quality on a system with nuclear option at peak load hours by incorporating two objective functions fuel cost and flat voltage profile. The fuel cost objective function is to minimize the system generation cost while flat voltage profile is to maximize power quality by minimizing voltage difference/variation. The study was conducted through literature study, determining objective functions, incorporating the objective functions, model simulation using case example and sensitivity analysis. The case example used is the IEEE 9 bus system that has been added fuel function of nuclear power plant (NPP), coal power plant (CPP), and gas turbine power plant (GTPP). ETAP12.6.0 software is used for the simulation. The sensitivity analysis was performed by changing the weighting value of each objective function. The results show that OPF will be reach 60 % weight on fuel costs and 40 % on flat voltage profile. The optimal generation cost is $ 7266/hour and the difference of maximum to minimum voltage is 2.85 %. CPP generates 33.2 MW + 22.1 MVar and GTPP 80.7 MW + 13.8 MVar. Meanwhile NPP generates 89.9 MW + 12.9 MVar and is economic when generating less than 90 MW. (author)

  17. Local and integral ulrasonic gauges for two-phase flow instrumentation in nuclear reactor and safety technology. Final report

    Energy Technology Data Exchange (ETDEWEB)

    Chochlov, V.N.; Duncev, A.V.; Ivanov, V.V.; Kontelev, V.V.; Melnikov, V.I.; Stoppel, L.K. [Technical State Univ. of Nishny Novgorod (Russian Federation); Prasser, H.M.; Zippe, W.; Zschau, J. [Forschungszentrum Rossendorf e.V. (FZR) (Germany). Inst. fuer Sicherheitsforschung; Zboray, R. [Technische Univ. Delft (Netherlands). Interfacultair Reactor Inst.

    2000-11-01

    The present project was executed in the frame of a co-operation agreement between FZR and the scientific group of Prof. Melnikov of the technical University of Nishny Novgorod (TUNN) in the Russian Federation. It is part of the Federal Government's programme for the provision of advice for Eastern Europe on the buildin