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

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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.

Flow cytometric evaluation of peripheral blood and bone marrow and fine-needle aspirate samples from multiple sites in dogs with multicentric lymphoma.

Science.gov (United States)

Joetzke, Alexa E; Eberle, Nina; Nolte, Ingo; Mischke, Reinhard; Simon, Daniela

2012-06-01

To determine whether the extent of disease in dogs with lymphoma can be assessed via flow cytometry and to evaluate the suitability of fine-needle aspirates from the liver and spleen of dogs for flow cytometric examination. 44 dogs with multicentric B-cell (n = 35) or T-cell lymphoma (9) and 5 healthy control dogs. Procedures-Peripheral blood and bone marrow samples and fine-needle aspirates of lymph node, liver, and spleen were examined via flow cytometry. Logarithmically transformed T-cell-to-B-cell percentage ratio (log[T:B]) values were calculated. Thresholds defined by use of log(T:B) values of samples from control dogs were used to determine extranodal lymphoma involvement in lymphoma-affected dogs; results were compared with cytologic findings. 12 of 245 (5%) samples (9 liver, 1 spleen, and 2 bone marrow) had insufficient cellularity for flow cytometric evaluation. Mean log(T:B) values of samples from dogs with B-cell lymphoma were significantly lower than those of samples from the same site in dogs with T-cell lymphoma and in control dogs. In dogs with T-cell lymphoma, the log(T:B) of lymph node, bone marrow, and spleen samples was significantly higher than in control dogs. Of 165 samples assessed for extranodal lymphoma involvement, 116 (70%) tested positive via flow cytometric analysis; results agreed with cytologic findings in 133 of 161 (83%) samples evaluated via both methods. Results suggested that flow cytometry may aid in detection of extranodal lymphoma involvement in dogs, but further research is needed. Most fine-needle aspirates of liver and spleen were suitable for flow cytometric evaluation.

Determination of chitin content in fungal cell wall: an alternative flow cytometric method.

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Costa-de-Oliveira, Sofia; Silva, Ana P; Miranda, Isabel M; Salvador, Alexandre; Azevedo, Maria M; Munro, Carol A; Rodrigues, Acácio G; Pina-Vaz, Cidália

2013-03-01

The conventional methods used to evaluate chitin content in fungi, such as biochemical assessment of glucosamine release after acid hydrolysis or epifluorescence microscopy, are low throughput, laborious, time-consuming, and cannot evaluate a large number of cells. We developed a flow cytometric assay, efficient, and fast, based on Calcofluor White staining to measure chitin content in yeast cells. A staining index was defined, its value was directly related to chitin amount and taking into consideration the different levels of autofluorecence. Twenty-two Candida spp. and four Cryptococcus neoformans clinical isolates with distinct susceptibility profiles to caspofungin were evaluated. Candida albicans clinical isolate SC5314, and isogenic strains with deletions in chitin synthase 3 (chs3Δ/chs3Δ) and genes encoding predicted GlycosylPhosphatidylInositol (GPI)-anchored proteins (pga31Δ/Δ and pga62Δ/Δ), were used as controls. As expected, the wild-type strain displayed a significant higher chitin content (P relationship between chitin content and antifungal drug susceptibility phenotype was found, an association was established between the paradoxical growth effect in the presence of high caspofungin concentrations and the chitin content. This novel flow cytometry protocol revealed to be a simple and reliable assay to estimate cell wall chitin content of fungi. Copyright © 2013 International Society for Advancement of Cytometry.

Clonal heterogeneity of small-cell anaplastic carcinoma of the lung demonstrated by flow-cytometric DNA analysis

DEFF Research Database (Denmark)

Vindeløv, L L; Hansen, H H; Christensen, I J

1980-01-01

Flow-cytometric DNA analysis yields information on ploidy and proliferative characteristics of a cell population. The analysis was implemented on small-cell anaplastic carcinoma of the lung using a rapid detergent technique for the preparation of fine-needle aspirates for DNA determination and a ...

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.

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

Flow cytometric techniques for detection of candidate cancer stem cell subpopulations in canine tumour models.

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Blacking, T M; Waterfall, M; Samuel, K; Argyle, D J

2012-12-01

The cancer stem cell (CSC) hypothesis proposes that tumour growth is maintained by a distinct subpopulation of 'CSC'. This study applied flow cytometric methods, reported to detect CSC in both primary and cultured cancer cells of other species, to identify candidate canine subpopulations. Cell lines representing diverse canine malignancies, and cells derived from spontaneous canine tumours, were evaluated for expression of stem cell-associated surface markers (CD34, CD44, CD117 and CD133) and functional properties [Hoecsht 33342 efflux, aldehyde dehydrogenase (ALDH) activity]. No discrete marker-defined subsets were identified within established cell lines; cells derived directly from spontaneous tumours demonstrated more heterogeneity, although this diminished upon in vitro culture. Functional assays produced variable results, suggesting context-dependency. Flow cytometric methods may be adopted to identify putative canine CSC. Whilst cell lines are valuable in assay development, primary cells may provide a more rewarding model for studying tumour heterogeneity in the context of CSC. However, it will be essential to fully characterize any candidate subpopulations to ensure that they meet CSC criteria. © 2011 Blackwell Publishing Ltd.

Vortex-dislodged cells from bone marrow trephine biopsy yield satisfactory results for flow cytometric immunophenotyping.

Science.gov (United States)

Bommannan, K; Sachdeva, M U S; Gupta, M; Bose, P; Kumar, N; Sharma, P; Naseem, S; Ahluwalia, J; Das, R; Varma, N

2016-10-01

A good bone marrow (BM) sample is essential in evaluating many hematologic disorders. An unsuccessful BM aspiration (BMA) procedure precludes a successful flow cytometric immunophenotyping (FCI) in most hematologic malignancies. Apart from FCI, most ancillary diagnostic techniques in hematology are less informative. We describe the feasibility of FCI in vortex-dislodged cell preparation obtained from unfixed trephine biopsy (TB) specimens. In pancytopenic patients and dry tap cases, routine diagnostic BMA and TB samples were complemented by additional trephine biopsies. These supplementary cores were immediately transferred into sterile tubes filled with phosphate-buffered saline, vortexed, and centrifuged. The cell pellet obtained was used for flow cytometric immunophenotyping. Of 7955 BMAs performed in 42 months, 34 dry tap cases were eligible for the study. Vortexing rendered a cell pellet in 94% of the cases (32 of 34), and FCI rendered a rapid diagnosis in 100% of the cases (32 of 32) where cell pellets were available. We describe an efficient procedure which could be effectively utilized in resource-limited centers and reduce the frequency of repeat BMA procedures. © 2016 John Wiley & Sons Ltd.

Flow cytometric and radioisotopic determinations of platelet survival time in normal cats and feline leukemia virus-infected cats

International Nuclear Information System (INIS)

Jacobs, R.M.; Boyce, J.T.; Kociba, G.J.

1986-01-01

This study demonstrates the potential usefulness of a flow cytometric technique to measure platelet survival time in cats utilizing autologous platelets labeled in vitro with fluorescein isothiocyanate (FITC). When compared with a 51Cr method, no significant differences in estimated survival times were found. Both the 51Cr and FITC-labeling procedures induced similar changes in platelet shape and collagen-induced aggregation. Platelets labeled with FITC had significantly greater volumes compared with those of glutaraldehyde-fixed platelets. These changes were primarily related to the platelet centrifugation and washing procedures rather than the labels themselves. This novel technique potentially has wide applicability to cell circulation time studies as flow cytometry equipment becomes more readily available. Problems with the technique are discussed. In a preliminary study of the platelet survival time in feline leukemia virus (FeLV)-infected cats, two of three cats had significantly reduced survival times using both flow cytometric and radioisotopic methods. These data suggest increased platelet turnover in FeLV-infected cats

Flow cytometric and radioisotopic determinations of platelet survival time in normal cats and feline leukemia virus-infected cats

Energy Technology Data Exchange (ETDEWEB)

Jacobs, R.M.; Boyce, J.T.; Kociba, G.J.

1986-01-01

This study demonstrates the potential usefulness of a flow cytometric technique to measure platelet survival time in cats utilizing autologous platelets labeled in vitro with fluorescein isothiocyanate (FITC). When compared with a 51Cr method, no significant differences in estimated survival times were found. Both the 51Cr and FITC-labeling procedures induced similar changes in platelet shape and collagen-induced aggregation. Platelets labeled with FITC had significantly greater volumes compared with those of glutaraldehyde-fixed platelets. These changes were primarily related to the platelet centrifugation and washing procedures rather than the labels themselves. This novel technique potentially has wide applicability to cell circulation time studies as flow cytometry equipment becomes more readily available. Problems with the technique are discussed. In a preliminary study of the platelet survival time in feline leukemia virus (FeLV)-infected cats, two of three cats had significantly reduced survival times using both flow cytometric and radioisotopic methods. These data suggest increased platelet turnover in FeLV-infected cats.

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

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

Immunologic status of children with thyroid cancer living near Chernobyl (flow cytometric and electron microscopic study)

International Nuclear Information System (INIS)

Zak, K.P.; Gruzov, M.A.; Bolshova, B.V.; Afanasyeva, V.V.; Shlyakhovenko, V.S.; Vishnevskaya, O.A.; Tronko, N.D.

1996-01-01

It hag been carded out a light, election microscopic and flow cytometric study of blood leukocyte of children with malignant tumors (papillary carcinoma) of thyroid gland who were living at the moment of the accident near Chernobyl. The results obtained point out the presence of some disturbances of immune status of these children

Quality control in the application of flow cytometric assays of genetic damage due to environmental contaminants

International Nuclear Information System (INIS)

McCreedy, C.D.; Jagoe, C.H.; Brisbin, I.L. Jr.; Wentworth, R.W.; Dallas, C.E.

1995-01-01

Clinical technologies, such as flow cytometry, are increasingly adopted by environmental toxicologists to identify resource damage associated with exposure to xenobiotics. One application of flow cytometry allows the rapid determination of the DNA content of large numbers of individual cells, and can be used to detect aneuploidy or other genetic abnormalities. The laboratory has used this methodology in studies of genetic toxicology of fish, birds, arid mammals exposed to organic pollutants, metals and radionuclides, However, without appropriate quality controls, false positive results and other artifacts can arise from sample handling and preparations, inter and intra-individual variations, instrument noise and other sources. The authors describe the routine measures this laboratory employs to maintain quality control of genomic DNA analysis, including the control of staining conditions, machine standardization, pulse-width doublet discrimination, and, in particular, the use of internal controls and the use of time as a cytometric parameter. Neglect of these controls can produce erroneous results, leading to conclusions of genetic abnormalities when none are present. Conversely, attention to these controls, routinely used in clinical settings, facilitates the interpretation of flow cytometric data and allows the application of this sensitive indicator of genotoxic effects to a variety of environmental problems

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

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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.

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

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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.

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

Science.gov (United States)

Rose, Jonathan A; Wanner, Nicholas; Cheong, Hoi I; Queisser, Kimberly; Barrett, Patrick; Park, Margaret; Hite, Corrine; Naga Prasad, Sathyamangla V; Erzurum, Serpil; Asosingh, Kewal

2016-01-01

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.

Novel flow cytometric analysis of the progress and route of internalization of a monoclonal anti-carcinoembryonic antigen (CEA) antibody.

Science.gov (United States)

Ford, C H; Tsaltas, G C; Osborne, P A; Addetia, K

1996-03-01

A flow cytometric method of studying the internalization of a monoclonal antibody (Mab) directed against carcinoembryonic antigen (CEA) has been compared with Western blotting, using three human colonic cancer cell lines which express varying amounts of the target antigen. Cell samples incubated for increasing time intervals with fluoresceinated or unlabelled Mab were analyzed using flow cytometry or polyacrylamide gel electrophoresis and Western blotting. SDS/PAGE analysis of cytosolic and membrane components of solubilized cells from the cell lines provided evidence of non-degraded internalized anti-CEA Mab throughout seven half hour intervals, starting at 5 min. Internalized anti-CEA was detected in the case of high CEA expressing cell lines (LS174T, SKCO1). Very similar results were obtained with an anti-fluorescein flow cytometric assay. Given that these two methods consistently provided comparable results, use of flow cytometry for the detection of internalized antibody is suggested as a rapid alternative to most currently used methods for assessing antibody internalization. The question of the endocytic route followed by CEA-anti-CEA complexes was addressed by using hypertonic medium to block clathrin mediated endocytosis.

Novel nuclei isolation buffer for flow cytometric genome size estimation of Zingiberaceae: a comparison with common isolation buffers.

Science.gov (United States)

Sadhu, Abhishek; Bhadra, Sreetama; Bandyopadhyay, Maumita

2016-11-01

Cytological parameters such as chromosome numbers and genome sizes of plants are used routinely for studying evolutionary aspects of polyploid plants. Members of Zingiberaceae show a wide range of inter- and intrageneric variation in their reproductive habits and ploidy levels. Conventional cytological study in this group of plants is severely hampered by the presence of diverse secondary metabolites, which also affect their genome size estimation using flow cytometry. None of the several nuclei isolation buffers used in flow cytometry could be used very successfully for members of Zingiberaceae to isolate good quality nuclei from both shoot and root tissues. The competency of eight nuclei isolation buffers was compared with a newly formulated buffer, MB01, in six different genera of Zingiberaceae based on the fluorescence intensity of propidium iodide-stained nuclei using flow cytometric parameters, namely coefficient of variation of the G 0 /G 1 peak, debris factor and nuclei yield factor. Isolated nuclei were studied using fluorescence microscopy and bio-scanning electron microscopy to analyse stain-nuclei interaction and nuclei topology, respectively. Genome contents of 21 species belonging to these six genera were determined using MB01. Flow cytometric parameters showed significant differences among the analysed buffers. MB01 exhibited the best combination of analysed parameters; photomicrographs obtained from fluorescence and electron microscopy supported the superiority of MB01 buffer over other buffers. Among the 21 species studied, nuclear DNA contents of 14 species are reported for the first time. Results of the present study substantiate the enhanced efficacy of MB01, compared to other buffers tested, in the generation of acceptable cytograms from all species of Zingiberaceae studied. Our study facilitates new ways of sample preparation for further flow cytometric analysis of genome size of other members belonging to this highly complex polyploid family

Flow cytometric life cycle analysis in cellular radiation biology

International Nuclear Information System (INIS)

Wood, J.C.S.

1982-01-01

Three approaches to flow cytometric histogram analysis were developed: (1) differential histogram analysis, (2) DNA histogram analysis, and (3) multiparameter data analysis. These techniques were applied to an important unresolved problem in radiation biology. The initial responses to irradiation of a mammalian cell which occur during the first two cell cycles following the irradiation are of considerable interest to the radiation biologist. During the first two post-irradiation cell cycles, cells which ultimately will survive repair radiation-induced damage, while some cells begin to express some of the radiation-induced nuclear and chomatin damage. Caffeine- and thymidine-treated, and untreated gamma-irradiated cell populations were studied with respect to the radiation-induced G2 delay, deficient DNA synthesis, and the appearance of cells with abnormal DNA contents. It is hypothesized that the measured deficiency in DNA synthesis observed in the first post-irradiation cell cycle may be a result of daughter cells from abnormal first post-irradiation mitoses

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

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

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

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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.

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.

Flow cytometric and morphological analyses of Pinus pinaster somatic embryogenesis.

Science.gov (United States)

Marum, Liliana; Loureiro, João; Rodriguez, Eleazar; Santos, Conceição; Oliveira, M Margarida; Miguel, Célia

2009-09-25

An approach combining morphological profiling and flow cytometric analysis was used to assess genetic stability during the several steps of somatic embryogenesis in Pinus pinaster. Embryogenic cell lines of P. pinaster were established from immature zygotic embryos excised from seeds obtained from open-pollinated trees. During the maturation stage, phenotype of somatic embryos was characterized as being either normal or abnormal. Based upon the prevalent morphological traits, different types of abnormal embryos underwent further classification and quantification. Nuclear DNA content of maritime pine using the zygotic embryos was estimated to be 57.04 pg/2C, using propidium iodide flow cytometry. According to the same methodology, no significant differences (P< or =0.01) in DNA ploidy were detected among the most frequently observed abnormal phenotypes, embryogenic cell lines, zygotic and normal somatic embryos, and somatic embryogenesis-derived plantlets. Although the differences in DNA ploidy level do not exclude the occurrence of a low level of aneuploidy, the results obtained point to the absence of major changes in ploidy level during the somatic embryogenesis process of this economically important species. Therefore, our primary goal of true-to-typeness was assured at this level.

Flow cytometric analysis of cell-surface and intracellular antigens in leukemia diagnosis.

Science.gov (United States)

Knapp, W; Strobl, H; Majdic, O

1994-12-15

New technology allows highly sensitive flow cytometric detection and quantitative analysis of intracellular antigens in normal and malignant hemopoietic cells. With this technology, the earliest stages of myeloid and lymphoid differentiation can easily and reliably be identified using antibodies directed against (pro-)myeloperoxidase/MPO, CD22 and CD3 antigens, respectively. Particularly for the analysis of undifferentiated acute myeloblastic leukemia (AML) cells, the immunological demonstration of intracellular MPO or its enzymatically inactive proforms is highly relevant, since other myeloid marker molecules such as CD33, CD13, or CDw65 are either not restricted to the granulomonocytic lineage or appear later in differentiation. By combining MPO staining with staining for lactoferrin (LF), undifferentiated cells can be distinguished from the granulomonocytic maturation compartment in bone marrow, since LF is selectively expressed from the myelocyte stage of differentiation onward. The list of informative intracellular antigens to be used in leukemia cell analysis will certainly expand in the near future. One candidate, intracellular CD68, has already been tested by us, and results are presented. Also dealt within this article are surface marker molecules not (as yet) widely used in leukemia cell analysis but with the potential to provide important additional information. Among them are the surface structures CD15, CD15s, CDw65, CD79a (MB-1), CD79b (B29), CD87 (uPA-R), and CD117 (c-kit).

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...

Flow cytometric osmotic fragility test and eosin-5'-maleimide dye-binding tests are better than conventional osmotic fragility tests for the diagnosis of hereditary spherocytosis.

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Arora, R D; Dass, J; Maydeo, S; Arya, V; Radhakrishnan, N; Sachdeva, A; Kotwal, J; Bhargava, M

2018-03-24

Hereditary spherocytosis (HS) is the most common inherited hemolytic anemia with heterogeneous clinico-laboratory manifestations. We evaluated the flow-cytometric tests: eosin-5'-maleimide (EMA) and flow-cytometric osmotic fragility test (FOFT) and the conventional osmotic fragility tests (OFT) for the diagnosis of hereditary spherocytosis (HS). One hundred two suspected HS patients underwent EMA, FOFT, incubated OFT (IOFT), and room temperature OFT (RT-OFT). In addition, 10 cases of immune hemolytic anemia (IHA) were included, and performance of the above 4 tests was evaluated. For EMA and FOFT, 5 normal controls were assessed together with the patients and cutoffs were calculated using receiver-operator-characteristics curve (ROC) analysis. The best cutoff for %EMA decrease was 12.5%, and for FOFT, %residual red cells (%RRC) was 25.6%. The sensitivity and specificity of RT-OFT was 62.06% and 86.3%, respectively, while that of IOFT was 79.31% and 87.67%, respectively. Both flow cytometric tests performed better. Sensitivity and specificity of EMA was 86.2% and 93.9% respectively, and that of FOFT was 96.6% and 98.63%, respectively. The combination of the FOFT with IOFT or EMA dye-binding test yields a sensitivity of 100%, but with EMA, it had a higher specificity. Hb/MCHC was a predictor of the severity of the disease while %EMA decrease and %RRC did not correlate with severity of the disease. Flow-cytometric osmotic fragility test is the best possible single test followed by EMA for diagnosis of HS. A combination of FOFT and EMA can correctly diagnose 100% patients. These tests are likely to replace conventional OFTs in future. © 2018 John Wiley & Sons Ltd.

A flow-cytometric gram-staining technique for milk-associated bacteria.

Science.gov (United States)

Holm, Claus; Jespersen, Lene

2003-05-01

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 degrees 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 degrees 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 degrees 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.

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

Cytometric analysis of irradiation damaged chromosomes

International Nuclear Information System (INIS)

Wilder, M.E.; Raju, M.R.

1982-01-01

Irradiation of cells in interphase results in dose-dependent damage to DNA which is discernable by flow-cytometric analysis of chromosomes. The quantity (and possibly the quality) of chromosomal changes is different in survival-matched doses of x and α irradiation. It may, therefore, be possible to use these methods for analysis of dose and type of exposure in unknown cases

Flow cytometric chemosensitivity assay using JC‑1, a sensor of mitochondrial transmembrane potential, in acute leukemia.

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Yokosuka, Tomoko; Goto, Hiroaki; Fujii, Hisaki; Naruto, Takuya; Takeuchi, Masanobu; Tanoshima, Reo; Kato, Hiromi; Yanagimachi, Masakatsu; Kajiwara, Ryosuke; Yokota, Shumpei

2013-12-01

The purpose of the study is to establish a simple and relatively inexpensive flow cytometric chemosensitivity assay (FCCA) for leukemia to distinguish leukemic blasts from normal leukocytes in clinical samples. We first examined whether the FCCA with the mitochondrial membrane depolarization sensor, 5, 50, 6, 60-tetrachloro-1, 10, 3, 30 tetraethyl benzimidazolo carbocyanine iodide (JC-1), could detect drug-induced apoptosis as the conventional FCCA by annexin V/7-AAD detection did and whether it was applicable in the clinical samples. Second, we compared the results of the FCCA for prednisolone (PSL) with clinical PSL response in 18 acute lymphoblastic leukemia (ALL) patients to evaluate the reliability of the JC-1 FCCA. Finally, we performed the JC-1 FCCA for bortezomib (Bor) in 25 ALL or 11 acute myeloid leukemia (AML) samples as the example of the clinical application of the FCCA. In ALL cells, the results of the JC-1 FCCA for nine anticancer drugs were well correlated with those of the conventional FCCA using anti-annexin V antibody (P < 0.001). In the clinical samples from 18 children with ALL, the results of the JC-1 FCCA for PSL were significantly correlated with the clinical PSL response (P = 0.005). In ALL samples, the sensitivity for Bor was found to be significantly correlated with the sensitivity for PSL (P = 0.005). In AML samples, the Bor sensitivity was strongly correlated with the cytarabine sensitivity (P = 0.0003). This study showed the reliability of a relatively simple and the FCCA using JC-1, and the possibility for the further clinical application.

Automated analysis of flow cytometric data for measuring neutrophil CD64 expression using a multi-instrument compatible probability state model.

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Wong, Linda; Hill, Beth L; Hunsberger, Benjamin C; Bagwell, C Bruce; Curtis, Adam D; Davis, Bruce H

2015-01-01

Leuko64™ (Trillium Diagnostics) is a flow cytometric assay that measures neutrophil CD64 expression and serves as an in vitro indicator of infection/sepsis or the presence of a systemic acute inflammatory response. Leuko64 assay currently utilizes QuantiCALC, a semiautomated software that employs cluster algorithms to define cell populations. The software reduces subjective gating decisions, resulting in interanalyst variability of state modeling (PSM). Four hundred and fifty-seven human blood samples were processed using the Leuko64 assay. Samples were analyzed on four different flow cytometer models: BD FACSCanto II, BD FACScan, BC Gallios/Navios, and BC FC500. A probability state model was designed to identify calibration beads and three leukocyte subpopulations based on differences in intensity levels of several parameters. PSM automatically calculates CD64 index values for each cell population using equations programmed into the model. GemStone software uses PSM that requires no operator intervention, thus totally automating data analysis and internal quality control flagging. Expert analysis with the predicate method (QuantiCALC) was performed. Interanalyst precision was evaluated for both methods of data analysis. PSM with GemStone correlates well with the expert manual analysis, r(2) = 0.99675 for the neutrophil CD64 index values with no intermethod bias detected. The average interanalyst imprecision for the QuantiCALC method was 1.06% (range 0.00-7.94%), which was reduced to 0.00% with the GemStone PSM. The operator-to-operator agreement in GemStone was a perfect correlation, r(2) = 1.000. Automated quantification of CD64 index values produced results that strongly correlate with expert analysis using a standard gate-based data analysis method. PSM successfully evaluated flow cytometric data generated by multiple instruments across multiple lots of the Leuko64 kit in all 457 cases. The probability-based method provides greater objectivity, higher

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.

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

Quantification of circulating mature endothelial cells using a whole blood four-color flow cytometric assay.

Science.gov (United States)

Jacques, Nathalie; Vimond, Nadege; Conforti, Rosa; Griscelli, Franck; Lecluse, Yann; Laplanche, Agnes; Malka, David; Vielh, Philippe; Farace, Françoise

2008-09-15

Circulating endothelial cells (CEC) are currently proposed as a potential biomarker for measuring the impact of anti-angiogenic treatments in cancer. However, the lack of consensus on the appropriate method of CEC measurement has led to conflicting data in cancer patients. A validated assay adapted for evaluating the clinical utility of CEC in large cohorts of patients undergoing anti-angiogenic treatments is needed. We developed a four-color flow cytometric assay to measure CEC as CD31(+), CD146(+), CD45(-), 7-amino-actinomycin-D (7AAD)(-) events in whole blood. The distinctive features of the assay are: (1) staining of 1 ml whole blood, (2) use of a whole blood IgPE control to measure accurately background noise, (3) accumulation of a large number of events (almost 5 10(6)) to ensure statistical analysis, and (4) use of 10 microm fluorescent microbeads to evaluate the event size. Assay reproducibility was determined in duplicate aliquots of samples drawn from 20 metastatic cancer patients. Assay linearity was tested by spiking whole blood with low numbers of HUVEC. Five-color flow cytometric experiments with CD144 were performed to confirm the endothelial origin of the cells. CEC were measured in 20 healthy individuals and 125 patients with metastatic cancer. Reproducibility was good between duplicate aliquots (r(2)=0.948, mean difference between duplicates of 0.86 CEC/ml). Detected HUVEC correlated with spiked HUVEC (r(2)=0.916, mean recovery of 100.3%). Co-staining of CD31, CD146 and CD144 confirmed the endothelial nature of cells identified as CEC. Median CEC levels were 6.5/ml (range, 0-15) in healthy individuals and 15.0/ml (range, 0-179) in patients with metastatic carcinoma (p<0.001). The assay proposed here allows reproducible and sensitive measurement of CEC by flow cytometry and could help evaluate CEC as biomarkers of anti-angiogenic therapies in large cohorts of patients.

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

Science.gov (United States)

Fröhling, Antje; Schlüter, Oliver

2015-01-01

Since heat sensitivity of fruits and vegetables limits the application of thermal inactivation processes, new emerging inactivation technologies have to be established to fulfill 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 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 parameters. PMID

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

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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

Assessment of Equine Autoimmune Thrombocytopenia (EAT by flow cytometry

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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.

Flow cytometric determination of radiation-induced chromosome damage and its correlation with cell survival

International Nuclear Information System (INIS)

Welleweerd, J.; Wilder, M.E.; Carpenter, S.G.; Raju, M.R.

1984-01-01

Chinese hamster M3-1 cells were irradiated with several doses of x rays or α particles from 238 Pu. Propidium iodide-stained chromosome suspensions were prepared at different times after irradiation; cells were also assayed for survival. The DNA histograms of these chromosomes showed increased background counts with increased doses of radiation. This increase in background was cell-cycle dependent and was correlated with cell survival. The correlation between radiation-induced chromosome damage and cell survival was the same for X rays and α particles. Data are presented which indicate that flow cytometric analysis of chromosomes of irradiated cell populations can be a useful adjunct to classical cytogenic analysis of irradiation-induced chromosomal damage by virtue of its ability to express and measure chromosomal damage not seen by classical cytogenic methods

Analytical validation of a flow cytometric protocol for quantification of platelet microparticles in dogs.

Science.gov (United States)

Cremer, Signe E; Krogh, Anne K H; Hedström, Matilda E K; Christiansen, Liselotte B; Tarnow, Inge; Kristensen, Annemarie T

2018-06-01

Platelet microparticles (PMPs) are subcellular procoagulant vesicles released upon platelet activation. In people with clinical diseases, alterations in PMP concentrations have been extensively investigated, but few canine studies exist. This study aims to validate a canine flow cytometric protocol for PMP quantification and to assess the influence of calcium on PMP concentrations. Microparticles (MP) were quantified in citrated whole blood (WB) and platelet-poor plasma (PPP) using flow cytometry. Anti-CD61 antibody and Annexin V (AnV) were used to detect platelets and phosphatidylserine, respectively. In 13 healthy dogs, CD61 + /AnV - concentrations were analyzed with/without a calcium buffer. CD61 + /AnV - , CD61 + /AnV + , and CD61 - /AnV + MP quantification were validated in 10 healthy dogs. The coefficient of variation (CV) for duplicate (intra-assay) and parallel (inter-assay) analyses and detection limits (DLs) were calculated. CD61 + /AnV - concentrations were higher in calcium buffer; 841,800 MP/μL (526,000-1,666,200) vs without; 474,200 MP/μL (278,800-997,500), P < .05. In WB, PMP were above DLs and demonstrated acceptable (<20%) intra-assay and inter-assay CVs in 9/10 dogs: 1.7% (0.5-8.9) and 9.0% (0.9-11.9), respectively, for CD61 + /AnV - and 2.4% (0.2-8.7) and 7.8% (0.0-12.8), respectively, for CD61 + /AnV + . Acceptable CVs were not seen for the CD61 - /AnV + MP. In PPP, quantifications were challenged by high inter-assay CV, overlapping DLs and hemolysis and lipemia interfered with quantification in 5/10 dogs. Calcium induced higher in vitro PMP concentrations, likely due to platelet activation. PMP concentrations were reliably quantified in WB, indicating the potential for clinical applications. PPP analyses were unreliable due to high inter-CV and DL overlap, and not obtainable due to hemolysis and lipemia interference. © 2018 American Society for Veterinary Clinical Pathology.

Automatic analysis of flow cytometric DNA histograms from irradiated mouse male germ cells

International Nuclear Information System (INIS)

Lampariello, F.; Mauro, F.; Uccelli, R.; Spano, M.

1989-01-01

An automatic procedure for recovering the DNA content distribution of mouse irradiated testis cells from flow cytometric histograms is presented. First, a suitable mathematical model is developed, to represent the pattern of DNA content and fluorescence distribution in the sample. Then a parameter estimation procedure, based on the maximum likelihood approach, is constructed by means of an optimization technique. This procedure has been applied to a set of DNA histograms relative to different doses of 0.4-MeV neutrons and to different time intervals after irradiation. In each case, a good agreement between the measured histograms and the corresponding fits has been obtained. The results indicate that the proposed method for the quantitative analysis of germ cell DNA histograms can be usefully applied to the study of the cytotoxic and mutagenic action of agents of toxicological interest such as ionizing radiations.18 references

Microscopic and flow cytometric study of micronuclei in iododeoxyuridine labelled cells irradiated with soft X-rays

International Nuclear Information System (INIS)

Ludwikow, G.; Staalnacke, C.G.; Johanson, K.J.; Sundell-Bergman, S.; Richter, S.; Swedish Univ. of Agricultural Sciences, Uppsala; Uppsala Univ.

1990-01-01

Iododeoxyuridine labelled (IUdR(+)) and unlabelled (IUdR(-)) CHO cells irradiated with 2 Gy of soft X-rays showed only minor differences in the kinetics of micronuclei formation during the first 20 hours postirradiation period. Between 20 to 40 hours, the IUdR(-) cells showed approximately a constant number of micronuclei while the number of micronuclei in IUdR(+) cells was still increasing. The frequency of micronuclei was higher in IUdR(+) cells compared to IUdR(-) cells at 24 hours after irradiation with various doses up to 4.0 Gy. Dose modifying factors were found to be 1.3 (microscopic evaluation) and 1.8 (flow cytometric evaluation). Flow cytometry with use of two parameters, fluorescence from propidium iodide and light scattering, seems to be a good tool to estimate the frequency of micronuclei in CHO cells in the dose range up to about 4 Gy. At higher doses perturbation of the cell cycle and the appearance of dying cells will influence the results. (orig.)

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

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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.

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...

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)

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...

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.

Evaluation of Prognostic Factors Following Flow-Cytometric DNA Analysis after Cytokeratin Labelling: I. Breast Cancer

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Pauline Wimberger

2002-01-01

Full Text Available In gynecologic oncology valid prognostic factors are necessary to estimate the course of disease and to define biologically similar subgroups for analysis of therapeutic efficacy. The presented study is a prospective study concerning prognostic significance of DNA ploidy and S‐phase fraction in breast cancer following enrichment of tumor cells by cytokeratin labelling. Epithelial cells were labeled by FITC‐conjugated cytokeratin antibody (CK 5, 6, 8, and CK 17 prior to flow cytometric cell cycle analysis in 327 fresh specimens of primary breast cancer. Univariate analysis in breast cancer detected the prognostic significance of DNA‐ploidy, S‐phase fraction and CV (coefficient of variation of G0G1‐peak of tumor cells for clinical outcome, especially for nodal‐negative patients. Multivariate analysis could not confirm prognostic evidence of DNA‐ploidy and S‐phase fraction. In conclusion, in breast cancer no clinical significance for determination of DNA‐parameters was found.

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.

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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.

Genome size variation among and within Camellia species by using flow cytometric analysis.

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Hui Huang

Full Text Available BACKGROUND: The genus Camellia, belonging to the family Theaceae, is economically important group in flowering plants. Frequent interspecific hybridization together with polyploidization has made them become taxonomically "difficult taxa". The DNA content is often used to measure genome size variation and has largely advanced our understanding of plant evolution and genome variation. The goals of this study were to investigate patterns of interspecific and intraspecific variation of DNA contents and further explore genome size evolution in a phylogenetic context of the genus. METHODOLOGY/PRINCIPAL FINDINGS: The DNA amount in the genus was determined by using propidium iodide flow cytometry analysis for a total of 139 individual plants representing almost all sections of the two subgenera, Camellia and Thea. An improved WPB buffer was proven to be suitable for the Camellia species, which was able to counteract the negative effects of secondary metabolite and generated high-quality results with low coefficient of variation values (CV <5%. Our results showed trivial effects on different tissues of flowers, leaves and buds as well as cytosolic compounds on the estimation of DNA amount. The DNA content of C. sinensis var. assamica was estimated to be 1C = 3.01 pg by flow cytometric analysis, which is equal to a genome size of about 2940 Mb. CONCLUSION: Intraspecific and interspecific variations were observed in the genus Camellia, and as expected, the latter was larger than the former. Our study suggests a directional trend of increasing genome size in the genus Camellia probably owing to the frequent polyploidization events.

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.

A Novel Tool for High-Throughput Screening of Granulocyte-Specific Antibodies Using the Automated Flow Cytometric Granulocyte Immunofluorescence Test (Flow-GIFT

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Xuan Duc Nguyen

2011-01-01

Full Text Available Transfusion-related acute lung injury (TRALI is a severe complication related with blood transfusion. TRALI has usually been associated with antibodies against leukocytes. The flow cytometric granulocyte immunofluorescence test (Flow-GIFT has been introduced for routine use when investigating patients and healthy blood donors. Here we describe a novel tool in the automation of the Flow-GIFT that enables a rapid screening of blood donations. We analyzed 440 sera from healthy female blood donors for the presence of granulocyte antibodies. As positive controls, 12 sera with known antibodies against anti-HNA-1a, -b, -2a; and -3a were additionally investigated. Whole-blood samples from HNA-typed donors were collected and the test cells isolated using cell sedimentation in a Ficoll density gradient. Subsequently, leukocytes were incubated with the respective serum and binding of antibodies was detected using FITC-conjugated antihuman antibody. 7-AAD was used to exclude dead cells. Pipetting steps were automated using the Biomek NXp Multichannel Automation Workstation. All samples were prepared in the 96-deep well plates and analyzed by flow cytometry. The standard granulocyte immunofluorescence test (GIFT and granulocyte agglutination test (GAT were also performed as reference methods. Sixteen sera were positive in the automated Flow-GIFT, while five of these sera were negative in the standard GIFT (anti—HNA 3a, n = 3; anti—HNA-1b, n = 1 and GAT (anti—HNA-2a, n = 1. The automated Flow-GIFT was able to detect all granulocyte antibodies, which could be only detected in GIFT in combination with GAT. In serial dilution tests, the automated Flow-GIFT detected the antibodies at higher dilutions than the reference methods GIFT and GAT. The Flow-GIFT proved to be feasible for automation. This novel high-throughput system allows an effective antigranulocyte antibody detection in a large donor population in order to prevent TRALI due to transfusion of

A novel tool for high-throughput screening of granulocyte-specific antibodies using the automated flow cytometric granulocyte immunofluorescence test (Flow-GIFT).

Science.gov (United States)

Nguyen, Xuan Duc; Dengler, Thomas; Schulz-Linkholt, Monika; Klüter, Harald

2011-02-03

Transfusion-related acute lung injury (TRALI) is a severe complication related with blood transfusion. TRALI has usually been associated with antibodies against leukocytes. The flow cytometric granulocyte immunofluorescence test (Flow-GIFT) has been introduced for routine use when investigating patients and healthy blood donors. Here we describe a novel tool in the automation of the Flow-GIFT that enables a rapid screening of blood donations. We analyzed 440 sera from healthy female blood donors for the presence of granulocyte antibodies. As positive controls, 12 sera with known antibodies against anti-HNA-1a, -b, -2a; and -3a were additionally investigated. Whole-blood samples from HNA-typed donors were collected and the test cells isolated using cell sedimentation in a Ficoll density gradient. Subsequently, leukocytes were incubated with the respective serum and binding of antibodies was detected using FITC-conjugated antihuman antibody. 7-AAD was used to exclude dead cells. Pipetting steps were automated using the Biomek NXp Multichannel Automation Workstation. All samples were prepared in the 96-deep well plates and analyzed by flow cytometry. The standard granulocyte immunofluorescence test (GIFT) and granulocyte agglutination test (GAT) were also performed as reference methods. Sixteen sera were positive in the automated Flow-GIFT, while five of these sera were negative in the standard GIFT (anti-HNA 3a, n = 3; anti-HNA-1b, n = 1) and GAT (anti-HNA-2a, n = 1). The automated Flow-GIFT was able to detect all granulocyte antibodies, which could be only detected in GIFT in combination with GAT. In serial dilution tests, the automated Flow-GIFT detected the antibodies at higher dilutions than the reference methods GIFT and GAT. The Flow-GIFT proved to be feasible for automation. This novel high-throughput system allows an effective antigranulocyte antibody detection in a large donor population in order to prevent TRALI due to transfusion of blood products.

Flow Cytometric DNA Analysis Using Cytokeratin Labeling for Identification of Tumor Cells in Carcinomas of the Breast and the Female Genital Tract

Directory of Open Access Journals (Sweden)

Rainer Kimmig

2001-01-01

Full Text Available Flow cytometric assessment of DNA‐ploidy and S‐phase fraction in malignant tumors is compromised by the heterogeneity of cell subpopulations derived from the malignant and surrounding connective tissue, e.g., tumor, stromal and inflammatory cells. To evaluate the effect on quality of DNA cell cycle analysis and determination of DNA ploidy, cytokeratin labeling of epithelial cells was used for tumor cell enrichment in breast, ovarian, cervical and endometrial cancer prior to DNA analysis. In a prospective study, tumor cell subpopulations of 620 malignant tumors were labeled by a FITC‐conjugated cytokeratin antibody (CK 5, 6, CK18 and CK 5, 6, 8 and CK 17, respectively prior to flow cytometric cell cycle analysis. Compared to total cell analysis, detection rate of DNA‐aneuploid tumors following cytokeratin labeling was increased from 62% to 76.5% in breast cancer, from 68% to 77% in ovarian cancer, from 60% to 80% in cervical cancer and from 30% to 53% in endometrial cancer. Predominantly in DNA‐diploid tumors, a significantly improved detection of S‐phase fraction of the tumor cells was shown due to the elimination of contaminating nonproliferating “normal cells”. S‐phase fraction following tumor cell enrichment was increased by 10% (mean following cytokeratin staining in ovarian and endometrial cancer, by 30% in breast cancer and even by 70% in cervical cancer compared to total cell analysis. Thus, diagnostic accuracy of DNA‐analysis was enhanced by cytokeratin labeling of tumor cells for all tumor entities investigated.

Multivariate analysis of flow cytometric data using decision trees

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Svenja eSimon

2012-04-01

Full Text Available 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's quality, we used stratified 5-fold 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.

Flow cytometric analysis of lectin binding to in vitro-cultured Perkinsus marinus surface carbohydrates

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Gauthier, J.D.; Jenkins, J.A.; La Peyre, Jerome F.

2004-01-01

Parasite surface glycoconjugates are frequently involved in cellular recognition and colonization of the host. This study reports on the identification of Perkinsus marinus surface carbohydrates by flow cytometric analyses of fluorescein isothiocyanate-conjugated lectin binding. Lectin-binding specificity was confirmed by sugar inhibition and Kolmogorov-Smirnov statistics. Clear, measurable fluorescence peaks were discriminated, and no parasite autofluorescence was observed. Parasites (GTLA-5 and Perkinsus-1 strains) harvested during log and stationary phases of growth in a protein-free medium reacted strongly with concanavalin A and wheat germ agglutinin, which bind to glucose-mannose and N-acetyl-D-glucosamine (GlcNAc) moieties, respectively. Both P. marinus strains bound with lower intensity to Maclura pomifera agglutinin, Bauhinia purpurea agglutinin, soybean agglutinin (N-acetyl-D-galactosamine-specific lectins), peanut agglutinin (PNA) (terminal galactose specific), and Griffonia simplicifolia II (GlcNAc specific). Only background fluorescence levels were detected with Ulex europaeus agglutinin I (L-fucose specific) and Limulus polyphemus agglutinin (sialic acid specific). The lectin-binding profiles were similar for the 2 strains except for a greater relative binding intensity of PNA for Perkinsus-1 and an overall greater lectin-binding capacity of Perkinsus-1 compared with GTLA-5. Growth stage comparisons revealed increased lectin-binding intensities during stationary phase compared with log phase of growth. This is the first report of the identification of surface glycoconjugates on a Perkinsus spp. by flow cytometry and the first to demonstrate that differential surface sugar expression is growth phase and strain dependent. ?? American Society of Parasitologists 2004.

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

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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.

Flow Cytometric Detection of PrPSc in Neurons and Glial Cells from Prion-Infected Mouse Brains.

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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

2006 Bethesda International Consensus recommendations on the immunophenotypic analysis of hematolymphoid neoplasia by flow cytometry: optimal reagents and reporting for the flow cytometric diagnosis of hematopoietic neoplasia.

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Wood, Brent L; Arroz, Maria; Barnett, David; DiGiuseppe, Joseph; Greig, Bruce; Kussick, Steven J; Oldaker, Teri; Shenkin, Mark; Stone, Elizabeth; Wallace, Paul

2007-01-01

Immunophenotyping by flow cytometry has become standard practice in the evaluation and monitoring of patients with hematopoietic neoplasia. However, despite its widespread use, considerable variability continues to exist in the reagents used for evaluation and the format in which results are reported. As part of the 2006 Bethesda Consensus conference, a committee was formed to attempt to define a consensus set of reagents suitable for general use in the diagnosis and monitoring of hematopoietic neoplasms. The committee included laboratory professionals from private, public, and university hospitals as well as large reference laboratories that routinely operate clinical flow cytometry laboratories with an emphasis on lymphoma and leukemia immunophenotyping. A survey of participants successfully identified the cell lineage(s) to be evaluated for each of a variety of specific medical indications and defined a set of consensus reagents suitable for the initial evaluation of each cell lineage. Elements to be included in the reporting of clinical flow cytometric results for leukemia and lymphoma evaluation were also refined and are comprehensively listed. The 2006 Bethesda Consensus conference represents the first successful attempt to define a set of consensus reagents suitable for the initial evaluation of hematopoietic neoplasia. Copyright 2007 Clinical Cytometry Society.

Flow cytometric analysis of immunoglobulin heavy chain expression in B-cell lymphoma and reactive lymphoid hyperplasia

Science.gov (United States)

Grier, David D; Al-Quran, Samer Z; Cardona, Diana M; Li, Ying; Braylan, Raul C

2012-01-01

The diagnosis of B-cell lymphoma (BCL) is often dependent on the detection of clonal immunoglobulin (Ig) light chain expression. In some BCLs, the determination of clonality based on Ig light chain restriction may be difficult. The aim of our study was to assess the utility of flow cytometric analysis of surface Ig heavy chain (HC) expression in lymphoid tissues in distinguishing lymphoid hyperplasias from BCLs, and also differentiating various BCL subtypes. HC expression on B-cells varied among different types of hyperplasias. In follicular hyperplasia, IgM and IgD expression was high in mantle cells while germinal center cells showed poor HC expression. In other hyperplasias, B cell compartments were blurred but generally showed high IgD and IgM expression. Compared to hyperplasias, BCLs varied in IgM expression. Small lymphocytic lymphomas had lower IgM expression than mantle cell lymphomas. Of importance, IgD expression was significantly lower in BCLs than in hyperplasias, a finding that can be useful in differentiating lymphoma from reactive processes. PMID:22400070

Flow cytometric kinetic assay of the activity of Na+/H+ antiporter in mammalian cells.

Science.gov (United States)

Dolz, María; O'Connor, José-Enrique; Lequerica, Juan L

2004-10-01

The Na(+)/H(+) exchanger (NHE) of mammalian cells is an integral membrane protein that extrudes H(+) ion in exchange for extracellular Na(+) and plays a crucial role in the regulation of intracellular pH (pHi). Thus, when pHi is lowered, NHE extrudes protons at a rate depending of pHi that can be expressed as pH units/s. To abolish the activity of other cellular pH-restoring systems, cells were incubated in bicarbonate-free Dulbecco's modified Eagle's medium buffered with HEPES. Flow cytometry was used to determine pHi with 2',7'-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein acetoxymethyl ester or 5-(and-6)-carboxy SNARF-1 acetoxymethyl ester acetate, and the appropriate fluorescence ratios were measured. The calibration of fluorescence ratios versus pHi was established by using ionophore nigericin. The activity of NHE was calculated by a kinetic flow cytometric assay as the slope at time 0 of the best-fit curve of pHi recovery versus time after intracellular acidification with a pulse of exogenous sodium propionate. The kinetic method allowed determination of the pHi-dependent activity of NHE in cell lines and primary cell cultures. NHE activity values were demonstrated to be up to 0.016 pH units/s within the pHi range of 7.3 to 6.3. The inhibition of NHE activity by the specific inhibitor ethyl isopropyl amiloride was easily detected by this method. The assay conditions can be used to relate variations in pHi with the activity of NHE and provide a standardized method to compare between different cells, inhibitors, models of ischemia by acidification, and other relevant experimental or clinical situations.

Flow cytometric detection of micronuclei by combined staining of DNA and membranes

International Nuclear Information System (INIS)

Wessels, J.M.; Nuesse, M.

1995-01-01

A new staining method is presented for flow cytometric measurement of micronuclei (MN) in cell cultures and human lymphocytes using membrane-specific fluorescent dyes in addition to DNA staining. Several combinations of fluorescent membrane and DNA dyes were studied for a better discrimination of MN from debris in a suspension of nuclei and micronuclei. For staining of membranes, the lipophilic dyes 2-hydroxyethyl-7,12,17-tris(methoxyethyl)porphycene (HEPn) and 1,6-diphenyl-1,3,5-hexatriene (DPH) were used in combination with ethidium bromide (EB), proflavine (PF), and Hoechst 33258 (HO). Due to their spectral properties, HO or EB combined with HEPn were not as suitable for the discrimination of MN from debris as was HEPn in combination with PF. With HEPn in combination with PF, however, additional noise was found at low fluorescence intensities, probably due to free fluorescent dye molecules in the solution. The optimal simultaneous staining of membranes and DNA was obtained using a combination of DPH and EB. The induction of MN in Chinese hamster and mouse NIH-3T3 cells by UV-B illumination was studied with this new staining technique. UV-B illumination (280-360 nm) induced MN in both cell lines. Chinese hamster cells were found to be more sensitive to these wavelengths. Illumination with wavelengths above 360 nm did not induce MN in either cell line. The results obtained from human lymphocytes using the combination of EB or DPH were comparable to the results obtained with the combination of EB and HO. 23 refs., 7 figs

Automation in high-content flow cytometry screening.

Science.gov (United States)

Naumann, U; Wand, M P

2009-09-01

High-content flow cytometric screening (FC-HCS) is a 21st Century technology that combines robotic fluid handling, flow cytometric instrumentation, and bioinformatics software, so that relatively large numbers of flow cytometric samples can be processed and analysed in a short period of time. We revisit a recent application of FC-HCS to the problem of cellular signature definition for acute graft-versus-host-disease. Our focus is on automation of the data processing steps using recent advances in statistical methodology. We demonstrate that effective results, on par with those obtained via manual processing, can be achieved using our automatic techniques. Such automation of FC-HCS has the potential to drastically improve diagnosis and biomarker identification.

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

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.

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.

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.

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.

Evaluation of flow cytometric HIT assays in relation to an IgG-Specific immunoassay and clinical outcome.

Science.gov (United States)

Kerényi, Adrienne; Beke Debreceni, Ildikó; Oláh, Zsolt; Ilonczai, Péter; Bereczky, Zsuzsanna; Nagy, Béla; Muszbek, László; Kappelmayer, János

2017-09-01

Heparin-induced thrombocytopenia (HIT) is a severe side effect of heparin treatment caused by platelet activating IgG antibodies generated against the platelet factor 4 (PF4)-heparin complex. Thrombocytopenia and thrombosis are the leading clinical symptoms of HIT. The clinical pretest probability of HIT was evaluated by the 4T score system. Laboratory testing of HIT was performed by immunological detection of antibodies against PF4-heparin complex (EIA) and two functional assays. Heparin-dependent activation of donor platelets by patient plasma was detected by flow cytometry. Increased binding of Annexin-V to platelets and elevated number of platelet-derived microparticles (PMP) were the indicators of platelet activation. EIA for IgG isotype HIT antibodies was performed in 405 suspected HIT patients. Based on negative EIA results, HIT was excluded in 365 (90%) of cases. In 40 patients with positive EIA test result functional tests were performed. Platelet activating antibodies were detected in 17 cases by Annexin V binding. PMP count analysis provided nearly identical results. The probability of a positive flow cytometric assay result was higher in patients with elevated antibody titer. 71% of patients with positive EIA and functional assay had thrombosis. EIA is an important first line laboratory test in the diagnosis of HIT; however, HIT must be confirmed by a functional test. Annexin V binding and PMP assays using flow cytometry are functional HIT tests convenient in a clinical diagnostic laboratory. The positive results of functional assays may predict the onset of thrombosis. © 2016 International Clinical Cytometry Society. © 2016 International Clinical Cytometry Society.

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

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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.

Cr(VI) induces DNA damage, cell cycle arrest and polyploidization: a flow cytometric and comet assay study in Pisum sativum.

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Rodriguez, Eleazar; Azevedo, Raquel; Fernandes, Pedro; Santos, Conceição

2011-07-18

Chromium(VI) is recognized as the most toxic valency of Cr, but its genotoxicity and cytostaticity in plants is still poorly studied. In order to analyze Cr(VI) cyto- and gentotoxicity, Pisum sativum L. plants were grown in soil and watered with solutions with different concentrations of Cr up to 2000 mg/L. After 28 days of exposure, leaves showed no significant variations in either cell cycle dynamics or ploidy level. As for DNA damage, flow cytometric (FCM) histograms showed significant differences in full peak coefficient of variation (FPCV) values, suggesting clastogenicity. This is paralleled by the Comet assay results, showing an increase in DNA damage for 1000 and 2000 mg/L. In roots, exposure to 2000 mg/L resulted in cell cycle arrest at the G(2)/M checkpoint. It was also verified that under the same conditions 40% of the individuals analyzed suffered polyploidization having both 2C and 4C levels. DNA damage analysis by the Comet assay and FCM revealed dose-dependent increases in DNA damage and FPCV. Through this, we have unequivocally demonstrated for the first time in plants that Cr exposure can result in DNA damage, cell cycle arrest, and polyploidization. Moreover, we critically compare the validity of the Comet assay and FCM in evaluating cytogenetic toxicity tests in plants and demonstrate that the data provided by both techniques complement each other and present high correlation levels. In conclusion, the data presented provides new insight on Cr effects in plants in general and supports the use of the parameters tested in this study as reliable endpoints for this metal toxicity in plants. © 2011 American Chemical Society

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

International Nuclear Information System (INIS)

Jensen, R.H.; Bigbee, W.L.; Langlois, R.G.; Grant, S.G.; Pleshanov, P.G.; Chirkov, A.A.; Pilinskaya, M.A.

1990-01-01

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 accident at Chernobyl, Ukraine, USSR, a number of people were exposed to whole body doses of ionizing 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. 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 refs., 5 figs

A Fast, Easy, and Customizable Eight-Color Flow Cytometric Method for Analysis of the Cellular Content of Bronchoalveolar Lavage Fluid in the Mouse.

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Daubeuf, François; Becker, Julien; Aguilar-Pimentel, Juan Antonio; Ebel, Claudine; Hrabě de Angelis, Martin; Hérault, Yann; Frossard, Nelly

2017-06-19

The cell composition of bronchoalveolar lavage fluid (BAL) is an important indicator of airway inflammation. It is commonly determined by cytocentrifuging leukocytes on slides, then staining, identifying, and counting them as eosinophils, neutrophils, macrophages, or lymphocytes according to morphological criteria under light microscopy, where it is not always easy to distinguish macrophages from lymphocytes. We describe here a one-step, easy-to-use, and easy-to-customize 8-color flow cytometric method for performing differential cell count and comparing it to morphological counts on stained cytospins. This method identifies BAL cells by a simultaneous one-step immunolabeling procedure using antibodies to identify T cells, B cells, neutrophils, eosinophils, and macrophages. Morphological analysis of flow-sorted cell subsets is used to validate this protocol. An important advantage of this basic flow cytometry protocol is the ability to customize it by the addition of antibodies to study receptor expression at leukocyte cell surfaces and identify subclasses of inflammatory cells as needed. © 2017 by John Wiley & Sons, Inc. Copyright © 2017 John Wiley & Sons, Inc.

Cytometric approaches to biological dosimetry

International Nuclear Information System (INIS)

Burger, G.

1983-01-01

Automatic cytometric techniques for detecting chromosomal aberrations are being tested but will not be used in routine examinations for some time to come. Automatic micronuclei counts are more promising but not sufficiently sensitive in the low dose range ( [de

Flow cytometric quantitation of phagocytosis in heparinized complete blood with latex particles and Candida albicans

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Jesús M. Egido

1997-12-01

Full Text Available We report a rapid method for the flow cytometric quantitation of phagocytosis in heparinized complete peripherial blood (HCPB, using commercially available phycoerythrin-conjugated latex particles of 1µm diameter. The method is faster and shows greater reproducibility than Bjerknes' (1984 standard technique using propidium iodide-stained Candida albicans, conventionally applied to the leukocytic layer of peripherial blood but here modified for HCPB. We also report a modification of Bjerknes' Intracellular Killing Test to allow its application to HCPB.Se da cuenta de un método rápido para la cuantización del flujo citométrico de la fagocitosis en sangre periférica completamente heparinizada (HCPB, mediante la utilización de partículas de látex phycoerythrin-conjugadas de 1µm de diámetro disponibles comercialmente. El método es más rápido y presenta mayor reproducibilidad que la técnica estandar de Bjerknes' (1984 utilizando propidium iodide-teñida Candida albicans, aplicada convencionalmente a la capa leucocitica de sangre periférica pero modificada por HCPB. Tambien damos cuenta de una modificación de Bjerknes' Intracellular Killing Test para permitir su aplicación a HCPB.

Simultaneous use of multiplex ligation-dependent probe amplification assay and flow cytometric DNA ploidy analysis in patients with acute leukemia.

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Reyes-Núñez, Virginia; Galo-Hooker, Evelyn; Pérez-Romano, Beatriz; Duque, Ricardo E; Ruiz-Arguelles, Alejandro; Garcés-Eisele, Javier

2018-01-01

The aim of this work was to simultaneously use multiplex ligation-dependent probe amplification (MLPA) assay and flow cytometric DNA ploidy analysis (FPA) to detect aneuploidy in patients with newly diagnosed acute leukemia. MLPA assay and propidium iodide FPA were used to test samples from 53 consecutive patients with newly diagnosed acute leukemia referred to our laboratory for immunophenotyping. Results were compared by nonparametric statistics. The combined use of both methods significantly increased the rate of detection of aneuploidy as compared to that obtained by each method alone. The limitations of one method are somehow countervailed by the other and vice versa. MPLA and FPA yield different yet complementary information concerning aneuploidy in acute leukemia. The simultaneous use of both methods might be recommended in the clinical setting. © 2017 International Clinical Cytometry Society. © 2017 International Clinical Cytometry Society.

Flow cytometry and integrated imaging

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V. Kachel

2000-06-01

Full Text Available It is a serious problem to relate the results of a flow cytometric analysis of a marine sample to different species. Images of particles selectively triggered by the flow cytometric analysis and picked out from the flowing stream give a valuable additional information on the analyzed organisms. The technical principles and problems of triggered imaging in flow are discussed, as well as the positioning of the particles in the plane of focus, freezing the motion of the quickly moving objects and what kinds of light sources are suitable for pulsed illumination. The images have to be stored either by film or electronically. The features of camera targets and the memory requirements for storing the image data and the conditions for the triggering device are shown. A brief explanation of the features of three realized flow cytometric imaging (FCI systems is given: the Macro Flow Planktometer built within the EUROMAR MAROPT project, the Imaging Module of the European Plankton Analysis System, supported by the MAST II EurOPA project and the most recently developed FLUVO VI universal flow cytometer including HBO 100- and laser excitation for fluorescence and scatter, Coulter sizing as well as bright field and and phase contrast FCI.

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.

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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.

Reliability-oriented multi-resource allocation in a stochastic-flow network

International Nuclear Information System (INIS)

Hsieh, C.-C.; Lin, M.-H.

2003-01-01

A stochastic-flow network consists of a set of nodes, including source nodes which supply various resources and sink nodes at which resource demands take place, and a collection of arcs whose capacities have multiple operational states. The network reliability of such a stochastic-flow network is the probability that resources can be successfully transmitted from source nodes through multi-capacitated arcs to sink nodes. Although the evaluation schemes of network reliability in stochastic-flow networks have been extensively studied in the literature, how to allocate various resources at source nodes in a reliable means remains unanswered. In this study, a resource allocation problem in a stochastic-flow network is formulated that aims to determine the optimal resource allocation policy at source nodes subject to given resource demands at sink nodes such that the network reliability of the stochastic-flow network is maximized, and an algorithm for computing the optimal resource allocation is proposed that incorporates the principle of minimal path vectors. A numerical example is given to illustrate the proposed algorithm

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

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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.

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)

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

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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.

A flow cytometric method for assessing viability of intraerythrocytic hemoparasites.

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Wyatt, C R; Goff, W; Davis, W C

1991-06-24

We have developed a rapid, reliable method of evaluating growth and viability of intraerythrocytic protozoan hemoparasites. The assay involves the selective uptake and metabolic conversion of hydroethidine to ethidium by live parasites present in intact erythrocytes. The red fluorescence imparted by ethidium intercalated into the DNA of the parasite permits the use of flow cytometry to distinguish infected erythrocytes with viable parasites from uninfected erythrocytes and erythrocytes containing dead parasites. Comparison of the fluorochromasia technique of enumerating the number and viability of hemoparasites in cultured erythrocytes with enumeration in Giemsa-stained films and uptake of [3H]hypoxanthine demonstrated the fluorochromasia technique yields comparable results. Studies with the hemoparasite, Babesia bovis, have shown the fluorochromasia technique can also be used to monitor the effect of parasiticidal drugs on parasites in vitro. The cumulative studies with the fluorochromasia assay suggest the assay will also prove useful in investigations focused on analysis of the immune response to hemoparasites and growth in vitro.

Ligand receptor dynamics at streptavidin-coated particle surfaces: A flow cytometric and spectrofluorimetric study

Energy Technology Data Exchange (ETDEWEB)

Buranda, T. [Univ. of New Mexico School of Medicine, Albuquerque, NM (United States)]|[Univ. of New Mexico, Albuquerque, NM (United States); Jones, G.M. [Univ. of New Mexico School of Medicine, Albuquerque, NM (United States); Nolan, J.P.; Keij, J. [Los Alamos National Labs., NM (United States); Lopez, G.P. [Univ. of New Mexico, Albuquerque, NM (United States); Sklar, L.A. [Univ. of New Mexico School of Medicine, Albuquerque, NM (United States)]|[Los Alamos National Lab., NM (United States)

1999-04-29

The authors have studied the binding of 5-((N-(5-(N-(6-(biotinoyl)amino)hexanoyl)amino)pentyl)thioureidyl)fluorescein (fluorescein biotin) to 6.2 {micro}m diameter, streptavidin-coated polystyrene beads using a combination of fluorimetric and flow cytometric methods. They have determined the average number of binding sites per bead, the extent of fluorescein quenching upon binding to the bead, and the association and dissociation kinetics. The authors estimate the site number to be {approx}1 million per bead. The binding of the fluorescein biotin ligand occurs in steps where the insertion of the biotin moiety into one receptor pocket is followed immediately by the capture of the fluorescein moiety by a neighboring binding pocket; fluorescence quenching is a consequence of this secondary binding. At high surface coverage, the dominant mechanism of quenching appears to be via the formation of nonfluorescent nearest-neighbor aggregates. At early times, the binding process is characterized by biphasic association and dissociation kinetics which are remarkably dependent on the initial concentration of the ligand. The rate constant for binding to the first receptor pocket of a streptavidin molecule is {approx}(1.3 {+-} 0.3) {times} 10{sup 7} 1{sup {minus}1} S{sup {minus}1}. The rate of binding of a second biotin may be reduced due to steric interference. The early time dissociative behavior is in sharp contrast to the typical stability associated with this system. The early time dissociative behavior is in sharp contrast to the typical stability associated with this system. The dissociation rate constant is as high as 0.05 s{sup {minus}1} shortly after binding, but decreases by 3 orders of magnitude after 3 h of binding. Potential sources for the time dependence of the dissociation rate constant are discussed.

Ploidy level variability in South American fescues (festuca L., poaceae): use of flow cytometry in up to 5 1/2-year-old caryopses and herbarium specimens.

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Smarda, P; Stancík, D

2006-01-01

Ploidy levels and chromosome numbers for 24 species of Festuca L. from 29 sites in Bolivia, Colombia, Ecuador and Venezuela are given. The ploidy level of 22 species is reported for the first time. A higher proportion of tetraploids in northern South America and the high frequency of polyploids in the whole continent are documented. In combination with chromosome counts, ploidy level was determined using flow cytometry in 4- to 5 1/2 -year-old herbarium specimens and mature caryopses. Flow cytometric determination from seeds was more reliable than determination from herbarium specimens. In herbarium specimens, the youngest, fresh green leaves, still hidden in sheaths, seem to be most suitable for cytometric determination. In old, brownish leaves, or poorly preserved herbarium specimens, the degradation of DNA signal in flow histograms was documented. DNA content measured in seeds was always higher than that measured in herbarium specimens, which may be caused by the presence of different cytosolic compounds. Differences of about 15% in relative DNA content of F. sodiroana and F. vaginalis was found in simultaneous measurements in seeds.

Predictive value of the flow cytometric PCNA - assay (proliferating cell nuclear antigen) in head and neck tumors after accelerated-hyperfractionated radiochemotherapy

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Wenz, F; Lohr, F; Rudat, V; Dietz, A; Flentje, M; Wannenmacher, M

1995-07-01

Purpose/Objective: Proliferation of surviving tumor cells during fractionated radiotherapy may limit tumor control, especially in rapidly proliferating tumors. It has been widely accepted, that this may play a major role in head and neck tumors. Several methods for the assessment of tumor proliferation have been developed, however, most of them are either laborious, invasive or potentially toxic. Today, the gold standard is the flow cytometric BrdUrd assay. We present a flow cytometric method for detection of PCNA, which is an intranuclear proliferation associated protein, in solid human head and neck tumors and how these data correlate with outcome. Materials and Methods: Pretherapeutic biopsies of 20 inoperable patients with squamous cell carcinoma of the head and neck (T3-4N2M0) were examined. The tissue was disaggregated with pepsin/HCl, antibody staining was performed using the clone PC10. Biparametric flow cytometry was performed after a FITC conjugated secondary antibody and propidiumjodine staining was applied. The PCNA-index (i.e. percentage PCNA-positive cells), the DNA-index and the S-phase fraction (SPF, euploid tumors only) were determined. The therapy consisted of combined accelerated-hyperfractionated radiochemotherapy (66 Gy in 5 wks, concomittant boost of 1.6 Gy/d in wks 4+5, Carboplatin in wks 1+5). The median follow-up time was 14 mths (5 - 28), the clinical partners (V.R., A.D.) were 'blinded' towards the PCNA-values. Results: 13 patients suffered from disease progession and 11 died. The actuarial median survival and disease free survival (DFS) were 14.4 and 10.7 mths, respectively. The PCNA-values ranged from 3.2 to 70% (median 9%), there were 7 aneuploid and 13 euploid tumors. SFP in the euploid tumors ranged from 4 to 14.5% (median 10.5%). Neither SFP nor ploidy had a significant influence on the outcome. The patients were divided according to their PCNA-value in higher (n=10) and lower (n=10) than the median. The survival and DFS were 13

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

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Polireddy, Kishore; Khan, Mohiuddin Md Taimur; Chavan, Hemantkumar; Young, Susan; Ma, Xiaochao; Waller, Anna; Garcia, Matthew; Perez, Dominique; Chavez, Stephanie; Strouse, Jacob J; Haynes, Mark K; Bologa, Cristian G; Oprea, Tudor I; Tegos, George P; Sklar, Larry A; Krishnamurthy, Partha

2012-01-01

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.

Measuring Dispositional Flow: Validity and reliability of the Dispositional Flow State Scale 2, Italian version.

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Riva, Eleonora F M; Riva, Giuseppe; Talò, Cosimo; Boffi, Marco; Rainisio, Nicola; Pola, Linda; Diana, Barbara; Villani, Daniela; Argenton, Luca; Inghilleri, Paolo

2017-01-01

The aim of this study is to evaluate the psychometric properties of the Italian version of the Dispositional Flow Scale-2 (DFS-2), for use with Italian adults, young adults and adolescents. In accordance with the guidelines for test adaptation, the scale has been translated with the method of back translation. The understanding of the item has been checked according to the latest standards on the culturally sensitive translation. The scale thus produced was administered to 843 individuals (of which 60.69% female), between the ages of 15 and 74. The sample is balanced between workers and students. The main activities defined by the subjects allow the sample to be divided into three categories: students, workers, athletes (professionals and semi-professionals). The confirmatory factor analysis, conducted using the Maximum Likelihood Estimator (MLM), showed acceptable fit indexes. Reliability and validity have been verified, and structural invariance has been verified on 6 categories of Flow experience and for 3 subsamples with different with different fields of action. Correlational analysis shows significant high values between the nine dimensions. Our data confirmed the validity and reliability of the Italian DFS-2 in measuring Flow experiences. The scale is reliable for use with Italian adults, young adults and adolescents. The Italian version of the scale is suitable for the evaluation of the subjective tendency to experience Flow trait characteristic in different contest, as sport, study and work.

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

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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".

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.

Long-term preservation of Tetraselmis indica (Chlorodendrophyceae, Chlorophyta) for flow cytometric analysis: Influence of fixative and storage temperature.

Science.gov (United States)

Naik, Sangeeta Mahableshwar; Anil, Arga Chandrashekar

2017-08-01

Immediate enumeration of phytoplankton is seldom possible. Therefore, fixation and subsequent storage are required for delayed analysis. This study investigated the influence of glutaraldehyde (GA) concentrations (0.25%, 0.5%, and 1%) and storage temperatures (-80°C LN2 , -80°C, -20°C, and 5°C) on Tetraselmis indica for flow cytometric analysis. Cell recovery, granularity, and membrane permeability were independent of GA concentration whereas cell size and chlorophyll autofluorescence were concentration dependent. After an initial cell loss (16-19%), no cell loss was observed when samples were stored at 5°C. Cell recovery was not influenced by storage temperature until 4months but later samples preserved at -80°C LN2 , -80°C, and -20°C resulted in ~41% cell loss. Although maximum cell recovery with minimal effect on cell integrity was obtained at 5°C, autofluorescence was retained better at -80°C LN2 and -80°C. This suggests that in addition to fixative, the choice of storage temperature is equally important. Thus for long-term preservation, especially to retain autofluorescence, the use of lower concentration (0.25%) of GA when stored at a lower temperature (-80°C LN2 and -80°C) while a higher concentration (1%) of GA when stored at a higher temperature (5°C) is recommended. Copyright © 2017 Elsevier B.V. All rights reserved.

[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%.

Deep sequencing and flow cytometric characterization of expanded effector memory CD8+CD57+ T cells frequently reveals T-cell receptor Vβ oligoclonality and CDR3 homology in acquired aplastic anemia.

Science.gov (United States)

Giudice, Valentina; Feng, Xingmin; Lin, Zenghua; Hu, Wei; Zhang, Fanmao; Qiao, Wangmin; Ibanez, Maria Del Pilar Fernandez; Rios, Olga; Young, Neal S

2018-05-01

Oligoclonal expansion of CD8 + CD28 - lymphocytes has been considered indirect evidence for a pathogenic immune response in acquired aplastic anemia. A subset of CD8 + CD28 - cells with CD57 expression, termed effector memory cells, is expanded in several immune-mediated diseases and may have a role in immune surveillance. We hypothesized that effector memory CD8 + CD28 - CD57 + cells may drive aberrant oligoclonal expansion in aplastic anemia. We found CD8 + CD57 + cells frequently expanded in the blood of aplastic anemia patients, with oligoclonal characteristics by flow cytometric Vβ usage analysis: skewing in 1-5 Vβ families and frequencies of immunodominant clones ranging from 1.98% to 66.5%. Oligoclonal characteristics were also observed in total CD8 + cells from aplastic anemia patients with CD8 + CD57 + cell expansion by T-cell receptor deep sequencing, as well as the presence of 1-3 immunodominant clones. Oligoclonality was confirmed by T-cell receptor repertoire deep sequencing of enriched CD8 + CD57 + cells, which also showed decreased diversity compared to total CD4 + and CD8 + cell pools. From analysis of complementarity-determining region 3 sequences in the CD8 + cell pool, a total of 29 sequences were shared between patients and controls, but these sequences were highly expressed in aplastic anemia subjects and also present in their immunodominant clones. In summary, expansion of effector memory CD8 + T cells is frequent in aplastic anemia and mirrors Vβ oligoclonal expansion. Flow cytometric Vβ usage analysis combined with deep sequencing technologies allows high resolution characterization of the T-cell receptor repertoire, and might represent a useful tool in the diagnosis and periodic evaluation of aplastic anemia patients. (Registered at clinicaltrials.gov identifiers: 00001620, 01623167, 00001397, 00071045, 00081523, 00961064 ). Copyright © 2018 Ferrata Storti Foundation.

Flow cytometric determination of micronucleus frequency.

Science.gov (United States)

Elhajouji, Azeddine; Lukamowicz-Rajska, Magdalena

2013-01-01

During the last two decades the micronucleus (MN) test has been extensively used as a genotoxicity screening tool of chemicals and in a variety of exploratory and mechanistic investigations. The MN is a biomarker for chromosomal damage or mitotic abnormalities, since it can originate from chromosome fragments or whole chromosomes that fail to be incorporated into daughter nuclei during mitosis (Fenech et al., Mutagenesis 26:125-132, 2011; Kirsch-Volders et al., Arch Toxicol 85:873-899, 2011). The simplicity of scoring, accuracy, amenability to automation by image analysis or flow cytometry, and readiness to be applied to a variety of cell types either in vitro or in vivo have made it a versatile tool that has contributed to a large extent in our understanding of key toxicological issues related to genotoxins and their effects at the cellular and organism levels. Recently, the final acceptance of the in vitro MN test guideline 487 (OECD Guideline for Testing of Chemicals, In vitro mammalian cell micronucleus test 487. In vitro mammalian cell micronucleus test (MNVIT). Organization for Economic Cooperation and Development, Paris, 2010) together with the standard in vivo MN test OECD guideline 474 (OECD Guideline for The Testing of Chemicals, Mammalian erythrocyte micronucleus test no. 474. Organization for Economic Cooperation and Development, Paris, 1997) will further position the assay as a key driver in the determination of the genotoxicity potential in exploratory research as well as in the regulatory environment. This chapter covers to some extent the protocol designs and experimental steps necessary for a successful performance of the MN test and an accurate analysis of the MN by the flow cytometry technique.

Reliability analysis of HVDC grid combined with power flow simulations

Energy Technology Data Exchange (ETDEWEB)

Yang, Yongtao; Langeland, Tore; Solvik, Johan [DNV AS, Hoevik (Norway); Stewart, Emma [DNV KEMA, Camino Ramon, CA (United States)

2012-07-01

Based on a DC grid power flow solver and the proposed GEIR, we carried out reliability analysis for a HVDC grid test system proposed by CIGRE working group B4-58, where the failure statistics are collected from literature survey. The proposed methodology is used to evaluate the impact of converter configuration on the overall reliability performance of the HVDC grid, where the symmetrical monopole configuration is compared with the bipole with metallic return wire configuration. The results quantify the improvement on reliability by using the later alternative. (orig.)

Evaluation of the reliability of transport networks based on the stochastic flow of moving objects

International Nuclear Information System (INIS)

Wu Weiwei; Ning, Angelika; Ning Xuanxi

2008-01-01

In transport networks, human beings are moving objects whose moving direction is stochastic in emergency situations. Based on this idea, a new model-stochastic moving network (SMN) is proposed. It is different from binary-state networks and stochastic-flow networks. The flow of SMNs has multiple-saturated states, that correspond to different flow values in each arc. In this paper, we try to evaluate the system reliability, defined as the probability that the saturated flow of the network is not less than a given demand d. Based on this new model, we obtain the flow probability distribution of every arc by simulation. An algorithm based on the blocking cutset of the SMN is proposed to evaluate the network reliability. An example is used to show how to calculate the corresponding reliabilities for different given demands of the SMN. Simulation experiments of different size were made and the system reliability precision was calculated. The precision of simulation results also discussed

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

Histopathologic and Flow-Cytometric Analysis of Neoplastic and Benign “background” Tissue in Breast Carcinoma Resections

Directory of Open Access Journals (Sweden)

Daniel W. Visscher

1998-01-01

Full Text Available Two-color, multiparametric synthesis phase fraction (SPF analysis of cytokeratin-labeled epithelial cells was flow cytometrically performed on both benign (SPFb and malignant tissue samples (if available, SPFt from 132 mastectomy/lumpectomy specimens. These data were then correlated with clinicopathologic features, including (1 tumor differentiation, (2 the proportion of tumor comprised of duct carcinoma-in situ (DCIS, and (3 the histology of accompanying benign breast tissue, classified by predominant microscopic pattern as intact, normal terminal duct lobular units (NTDLU, 34% of cases, atrophic (AT, 33% of cases, proliferative fibrocystic (PFC, 26% of cases, and non-proliferative fibrocystic (NPFC, 7% of cases. SPFt was inversely correlated with extent of DCIS (DCIS =0 – 20% tumor volume – 12.7% mean SPFt, vs. DCIS >20% tumor volume – 6.4% mean SPFt, p = 0.001. SPFt also correlated with the histology of background benign breast tissue (NTDLU – 14.8% mean SPFt vs. AT – 6.9% mean SPFt vs. PFC – 12.7% mean SPFt, p = 0.05 but it did not correlate with patient age or SPFb (overall mean =0.73%. SPFb was correlated with patient age (>56 yr – 0.59% mean SPFb vs. < yr – 0.84% mean SPFb, p = 0.02, with background histology (NTDLU – 1.1% mean SPFb vs. AT – 0.43% mean SPFb vs. PFC – 0.70% mean SPFb, p < 0.02 and with the grade of the neoplasm (well/moderate – 0.58% mean vs. poorly differentiated – 0.85% mean, p = 0.04. Patients having a background of PFC were significantly older than patients with a background of NTDLU (45.2 yr vs. 60.2 yr, p = 0.01.

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.

Predicting Flow Breakdown Probability and Duration in Stochastic Network Models: Impact on Travel Time Reliability

Energy Technology Data Exchange (ETDEWEB)

Dong, Jing [ORNL; Mahmassani, Hani S. [Northwestern University, Evanston

2011-01-01

This paper proposes a methodology to produce random flow breakdown endogenously in a mesoscopic operational model, by capturing breakdown probability and duration. Based on previous research findings that probability of flow breakdown can be represented as a function of flow rate and the duration can be characterized by a hazard model. By generating random flow breakdown at various levels and capturing the traffic characteristics at the onset of the breakdown, the stochastic network simulation model provides a tool for evaluating travel time variability. The proposed model can be used for (1) providing reliability related traveler information; (2) designing ITS (intelligent transportation systems) strategies to improve reliability; and (3) evaluating reliability-related performance measures of the system.

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)

Reliability-based assessment of flow assurance of hot waxy crude pipelines

Energy Technology Data Exchange (ETDEWEB)

Jinjun, Zhang; Wenke, Zhang; Jianlin, Ding; Bo, Yu [China University of Petroleum - Beijing (CUPB), Beijing (China)

2009-07-01

Waxy crude is characterized by its high pour point. Pipeline blockage may occur after prolonged shutdown of a pipeline due to crude oil gelation. Another flow assurance problem is the instable operation at a flow rate below the lowest allowable operation flow rate which is dependent on heat transfer of the pipeline and the viscosity vs. temperature relation of the crude pumped. Besides, for pipelines with thick wax deposition layer, massive depletion of wax deposit in some cases such as pipeline restart at high expelling pressure may also result in blockage of the pipeline, and the pig may be jammed during pigging as a result of thick wax deposition. Conventionally, assessment of these risks has been made by using the deterministic approach. However, many related physical quantities are subject to uncertainty and contribute to reliability of flow assurance. Therefore, the probabilistic approach is suggested and a framework of reliability based assessment of flow assurance of waxy crude pipelines is proposed in this paper. Discussions are also made on the limit state functions and target safety level. In the future study, development of an efficient and robust stochastic-numerical method is crucial. (author)

Development of the GO-FLOW reliability analysis methodology for nuclear reactor system

International Nuclear Information System (INIS)

Matsuoka, Takeshi; Kobayashi, Michiyuki

1994-01-01

Probabilistic Safety Assessment (PSA) is important in the safety analysis of technological systems and processes, such as, nuclear plants, chemical and petroleum facilities, aerospace systems. Event trees and fault trees are the basic analytical tools that have been most frequently used for PSAs. Several system analysis methods can be used in addition to, or in support of, the event- and fault-tree analysis. The need for more advanced methods of system reliability analysis has grown with the increased complexity of engineered systems. The Ship Research Institute has been developing a new reliability analysis methodology, GO-FLOW, which is a success-oriented system analysis technique, and is capable of evaluating a large system with complex operational sequences. The research has been supported by the special research fund for Nuclear Technology, Science and Technology Agency, from 1989 to 1994. This paper describes the concept of the Probabilistic Safety Assessment (PSA), an overview of various system analysis techniques, an overview of the GO-FLOW methodology, the GO-FLOW analysis support system, procedure of treating a phased mission problem, a function of common cause failure analysis, a function of uncertainty analysis, a function of common cause failure analysis with uncertainty, and printing out system of the results of GO-FLOW analysis in the form of figure or table. Above functions are explained by analyzing sample systems, such as PWR AFWS, BWR ECCS. In the appendices, the structure of the GO-FLOW analysis programs and the meaning of the main variables defined in the GO-FLOW programs are described. The GO-FLOW methodology is a valuable and useful tool for system reliability analysis, and has a wide range of applications. With the development of the total system of the GO-FLOW, this methodology has became a powerful tool in a living PSA. (author) 54 refs

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.

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

Go-flow: a reliability analysis methodology applicable to piping system

International Nuclear Information System (INIS)

Matsuoka, T.; Kobayashi, M.

1985-01-01

Since the completion of the Reactor Safety Study, the use of probabilistic risk assessment technique has been becoming more widespread in the nuclear community. Several analytical methods are used for the reliability analysis of nuclear power plants. The GO methodology is one of these methods. Using the GO methodology, the authors performed a reliability analysis of the emergency decay heat removal system of the nuclear ship Mutsu, in order to examine its applicability to piping systems. By this analysis, the authors have found out some disadvantages of the GO methodology. In the GO methodology, the signal is on-to-off or off-to-on signal, therefore the GO finds out the time point at which the state of a system changes, and can not treat a system which state changes as off-on-off. Several computer runs are required to obtain the time dependent failure probability of a system. In order to overcome these disadvantages, the authors propose a new analytical methodology: GO-FLOW. In GO-FLOW, the modeling method (chart) and the calculation procedure are similar to those in the GO methodology, but the meaning of signal and time point, and the definitions of operators are essentially different. In the paper, the GO-FLOW methodology is explained and two examples of the analysis by GO-FLOW are given

Alternative approach to automated management of load flow in engineering networks considering functional reliability

Directory of Open Access Journals (Sweden)

Ирина Александровна Гавриленко

2016-02-01

Full Text Available The approach to automated management of load flow in engineering networks considering functional reliability was proposed in the article. The improvement of the concept of operational and strategic management of load flow in engineering networks was considered. The verbal statement of the problem for thesis research is defined, namely, the problem of development of information technology for exact calculation of the functional reliability of the network, or the risk of short delivery of purpose-oriented product for consumers

Application of system reliability analytical method, GO-FLOW

International Nuclear Information System (INIS)

Matsuoka, Takeshi; Fukuto, Junji; Mitomo, Nobuo; Miyazaki, Keiko; Matsukura, Hiroshi; Kobayashi, Michiyuki

1999-01-01

The Ship Research Institute proceed a developmental study on GO-FLOW method with various advancing functionalities for the system reliability analysis method occupying main parts of PSA (Probabilistic Safety Assessment). Here was attempted to intend to upgrade functionality of the GO-FLOW method, to develop an analytical function integrated with dynamic behavior analytical function, physical behavior and probable subject transfer, and to prepare a main accident sequence picking-out function. In 1997 fiscal year, in dynamic event-tree analytical system, an analytical function was developed by adding dependency between headings. In simulation analytical function of the accident sequence, main accident sequence of MRX for improved ship propulsion reactor became possible to be covered perfectly. And, input data for analysis was prepared with a function capable easily to set by an analysis operator. (G.K.)

Fluorescence-intensity multiplexing: simultaneous seven-marker, two-color immunophenotyping using flow cytometry.

Science.gov (United States)

Bradford, Jolene A; Buller, Gayle; Suter, Michael; Ignatius, Michael; Beechem, Joseph M

2004-10-01

Conventional immuno-based multiparameter flow cytometric analysis has been limited by the requirement of a dedicated detection channel for each antibody-fluorophore set. To address the need to resolve multiple biological targets simultaneously, flow cytometers with as many as 10-15 detection channels have been developed. In this study, a new Zenon immunolabeling technology is developed that allows for multiple antigen detection per detection channel using a single fluorophore, through a unique method of fluorescence-intensity multiplexing. By varying the Zenon labeling reagent-to-antibody molar ratio, the fluorescence intensity of the antibody-labeled cellular targets can be used as a unique identifier. Although demonstrated in the present study with lymphocyte immunophenotyping, this approach is broadly applicable for any immuno-based multiplexed flow cytomety assay. Lymphocyte immunophenotyping of 38 clinical blood specimens using CD3, CD4, CD8, CD16, CD56, CD19, and CD20 antibodies was performed using conventional flow cytometric analysis and fluorescence-intensity multiplexing analysis. Conventional analysis measures a single antibody-fluorophore per photomultiplier tube (PMT). Fluorescence-intensity multiplex analysis simultaneously measures seven markers with two PMTs, using Zenon labeling reagent-antibody complexes in a single tube: CD19, CD4, CD8, and CD16 antibodies labeled with Zenon Alexa Fluor 488 Mouse IgG(1) labeling reagent and CD56, CD3, and CD20 antibodies labeled with Zenon R-Phycoerythrin (R-PE) Mouse IgG(1) or IgG(2b) labeling reagents. The lymphocyte immunophenotyping results from fluorescence-intensity multiplexing using Zenon labeling reagents in a single tube were comparable to results from conventional flow cytometric analysis. Simultaneous evaluation of multiple antigens using a single fluorophore can be performed using antibodies labeled with varying ratios of a Zenon labeling reagent. Labeling two sets of antibodies with different Zenon

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

Reliability Approach of a Compressor System using Reliability Block ...

African Journals Online (AJOL)

pc

2018-03-05

Mar 5, 2018 ... This paper presents a reliability analysis of such a system using reliability ... Keywords-compressor system, reliability, reliability block diagram, RBD .... the same structure has been kept with the three subsystems: air flow, oil flow and .... and Safety in Engineering Design", Springer, 2009. [3] P. O'Connor ...

Performance of the flow cytometric E-screen assay in screening estrogenicity of pure compounds and environmental samples

Energy Technology Data Exchange (ETDEWEB)

Vanparys, Caroline, E-mail: caroline.vanparys@ua.ac.be [Laboratory of Ecophysiology, Biochemistry and Toxicology, University of Antwerp, Antwerp (Belgium); Depiereux, Sophie; Nadzialek, Stephanie [Research Unit in Organismal Biology (URBO), University of Namur (FUNDP), Namur (Belgium); Robbens, Johan; Blust, Ronny [Laboratory of Ecophysiology, Biochemistry and Toxicology, University of Antwerp, Antwerp (Belgium); Kestemont, Patrick [Research Unit in Organismal Biology (URBO), University of Namur (FUNDP), Namur (Belgium); De Coen, Wim [Laboratory of Ecophysiology, Biochemistry and Toxicology, University of Antwerp, Antwerp (Belgium); European Chemicals Agency (ECHA), Helsinki (Finland)

2010-09-15

In vitro estrogenicity screens are believed to provide a first prioritization step in hazard characterization of endocrine disrupting chemicals. When applied to complex environmental matrices or mixture samples, they have been indicated valuable in estimating the overall estrogen-mimicking load. In this study, the performance of an adapted format of the classical E-screen or MCF-7 cell proliferation assay was profoundly evaluated to rank pure compounds as well as influents and effluents of sewage treatment plants (STPs) according to estrogenic activity. In this adapted format, flow cytometric cell cycle analysis was used to allow evaluation of the MCF-7 cell proliferative effects after only 24 h of exposure. With an average EC{sub 50} value of 2 pM and CV of 22%, this assay appears as a sensitive and reproducible system for evaluation of estrogenic activity. Moreover, estrogenic responses of 17 pure compounds corresponded well, qualitatively and quantitatively, with other in vitro and in vivo estrogenicity screens, such as the classical E-screen (R{sup 2} = 0.98), the estrogen receptor (ER) binding (R{sup 2} = 0.84) and the ER transcription activation assay (R{sup 2} = 0.87). To evaluate the applicability of this assay for complex samples, influents and effluents of 10 STPs covering different treatment processes, were compared and ranked according to estrogenic removal efficiencies. Activated sludge treatment with phosphorus and nitrogen removal appeared most effective in eliminating estrogenic activity, followed by activated sludge, lagoon and filter bed. This is well in agreement with previous findings based on chemical analysis or biological activity screens. Moreover, ER blocking experiments indicated that cell proliferative responses were mainly ER mediated, illustrating that the complexity of the end point, cell proliferation, compared to other ER screens, does not hamper the interpretation of the results. Therefore, this study, among other E-screen studies

Performance of the flow cytometric E-screen assay in screening estrogenicity of pure compounds and environmental samples

International Nuclear Information System (INIS)

Vanparys, Caroline; Depiereux, Sophie; Nadzialek, Stephanie; Robbens, Johan; Blust, Ronny; Kestemont, Patrick; De Coen, Wim

2010-01-01

In vitro estrogenicity screens are believed to provide a first prioritization step in hazard characterization of endocrine disrupting chemicals. When applied to complex environmental matrices or mixture samples, they have been indicated valuable in estimating the overall estrogen-mimicking load. In this study, the performance of an adapted format of the classical E-screen or MCF-7 cell proliferation assay was profoundly evaluated to rank pure compounds as well as influents and effluents of sewage treatment plants (STPs) according to estrogenic activity. In this adapted format, flow cytometric cell cycle analysis was used to allow evaluation of the MCF-7 cell proliferative effects after only 24 h of exposure. With an average EC 50 value of 2 pM and CV of 22%, this assay appears as a sensitive and reproducible system for evaluation of estrogenic activity. Moreover, estrogenic responses of 17 pure compounds corresponded well, qualitatively and quantitatively, with other in vitro and in vivo estrogenicity screens, such as the classical E-screen (R 2 = 0.98), the estrogen receptor (ER) binding (R 2 = 0.84) and the ER transcription activation assay (R 2 = 0.87). To evaluate the applicability of this assay for complex samples, influents and effluents of 10 STPs covering different treatment processes, were compared and ranked according to estrogenic removal efficiencies. Activated sludge treatment with phosphorus and nitrogen removal appeared most effective in eliminating estrogenic activity, followed by activated sludge, lagoon and filter bed. This is well in agreement with previous findings based on chemical analysis or biological activity screens. Moreover, ER blocking experiments indicated that cell proliferative responses were mainly ER mediated, illustrating that the complexity of the end point, cell proliferation, compared to other ER screens, does not hamper the interpretation of the results. Therefore, this study, among other E-screen studies, supports the use of

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

Flow cytometric assessment of viability of lactic acid bacteria

NARCIS (Netherlands)

Bunthof, C.J.; Bloemen, K.; Breeuwer, P.; Rombouts, F.M.; Abee, T.

2001-01-01

The viability of lactic acid bacteria is crucial for their applications as dairy starters and as probiotics. We investigated the usefulness of flow cytometry (FCM) for viability assessment of lactic acid bacteria. The esterase substrate carboxyfluorescein diacetate (cFDA) and the dye exclusion DNA

Quantitative dynamic reliability evaluation of AP1000 passive safety systems by using FMEA and GO-FLOW methodology

International Nuclear Information System (INIS)

Hashim Muhammad; Yoshikawa, Hidekazu; Matsuoka, Takeshi; Yang Ming

2014-01-01

The passive safety systems utilized in advanced pressurized water reactor (PWR) design such as AP1000 should be more reliable than that of active safety systems of conventional PWR by less possible opportunities of hardware failures and human errors (less human intervention). The objectives of present study are to evaluate the dynamic reliability of AP1000 plant in order to check the effectiveness of passive safety systems by comparing the reliability-related issues with that of active safety systems in the event of the big accidents. How should the dynamic reliability of passive safety systems properly evaluated? And then what will be the comparison of reliability results of AP1000 passive safety systems with the active safety systems of conventional PWR. For this purpose, a single loop model of AP1000 passive core cooling system (PXS) and passive containment cooling system (PCCS) are assumed separately for quantitative reliability evaluation. The transient behaviors of these passive safety systems are taken under the large break loss-of-coolant accident in the cold leg. The analysis is made by utilizing the qualitative method failure mode and effect analysis in order to identify the potential failure mode and success-oriented reliability analysis tool called GO-FLOW for quantitative reliability evaluation. The GO-FLOW analysis has been conducted separately for PXS and PCCS systems under the same accident. The analysis results show that reliability of AP1000 passive safety systems (PXS and PCCS) is increased due to redundancies and diversity of passive safety subsystems and components, and four stages automatic depressurization system is the key subsystem for successful actuation of PXS and PCCS system. The reliability results of PCCS system of AP1000 are more reliable than that of the containment spray system of conventional PWR. And also GO-FLOW method can be utilized for reliability evaluation of passive safety systems. (author)

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

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.

Application case study of AP1000 automatic depressurization system (ADS) for reliability evaluation by GO-FLOW methodology

Energy Technology Data Exchange (ETDEWEB)

Hashim, Muhammad, E-mail: hashimsajid@yahoo.com; Hidekazu, Yoshikawa, E-mail: yosikawa@kib.biglobe.ne.jp; Takeshi, Matsuoka, E-mail: mats@cc.utsunomiya-u.ac.jp; Ming, Yang, E-mail: myang.heu@gmail.com

2014-10-15

Highlights: • Discussion on reasons why AP1000 equipped with ADS system comparatively to PWR. • Clarification of full and partial depressurization of reactor coolant system by ADS system. • Application case study of four stages ADS system for reliability evaluation in LBLOCA. • GO-FLOW tool is capable to evaluate dynamic reliability of passive safety systems. • Calculated ADS reliability result significantly increased dynamic reliability of PXS. - Abstract: AP1000 nuclear power plant (NPP) utilized passive means for the safety systems to ensure its safety in events of transient or severe accidents. One of the unique safety systems of AP1000 to be compared with conventional PWR is the “four stages Automatic Depressurization System (ADS)”, and ADS system originally works as an active safety system. In the present study, authors first discussed the reasons of why four stages ADS system is added in AP1000 plant to be compared with conventional PWR in the aspect of reliability. And then explained the full and partial depressurization of RCS system by four stages ADS in events of transient and loss of coolant accidents (LOCAs). Lastly, the application case study of four stages ADS system of AP1000 has been conducted in the aspect of reliability evaluation of ADS system under postulated conditions of full RCS depressurization during large break loss of a coolant accident (LBLOCA) in one of the RCS cold legs. In this case study, the reliability evaluation is made by GO-FLOW methodology to determinate the influence of ADS system in dynamic reliability of passive core cooling system (PXS) of AP1000, i.e. what will happen if ADS system fails or successfully actuate. The GO-FLOW is success-oriented reliability analysis tool and is capable to evaluating the systems reliability/unavailability alternatively to Fault Tree Analysis (FTA) and Event Tree Analysis (ETA) tools. Under these specific conditions of LBLOCA, the GO-FLOW calculated reliability results indicated

Flow cytometry detection of planktonic cells with polycyclic aromatic hydrocarbons sorbed to cell surfaces

KAUST Repository

Cerezo, Maria I.; Linden, Matthew; Agusti, Susana

2017-01-01

Polycyclic aromatic hydrocarbons are very important components of oil pollution. These pollutants tend to sorb to cell surfaces, exerting toxic effects on organisms. Our study developed a flow cytometric method for the detection of PAHs sorbed

Flow cytometric applications of tumor biology: prospects and pitfalls

International Nuclear Information System (INIS)

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

Flow cytometric applications to tumour biology: prospects and pitfalls

International Nuclear Information System (INIS)

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

1980-01-01

A brief review of cytometry instrumentation and its potential applications in tumour biology is presented. DNA distribution measurements of cells from spontaneous dog tumours 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, however, to utilize the sophisticated developments in flow cytometry instrumentation. (author)

Automated flow cytometric analysis across large numbers of samples and cell types.

Science.gov (United States)

Chen, Xiaoyi; Hasan, Milena; Libri, Valentina; Urrutia, Alejandra; Beitz, Benoît; Rouilly, Vincent; Duffy, Darragh; Patin, Étienne; Chalmond, Bernard; Rogge, Lars; Quintana-Murci, Lluis; Albert, Matthew L; Schwikowski, Benno

2015-04-01

Multi-parametric flow cytometry is a key technology for characterization of immune cell phenotypes. However, robust high-dimensional post-analytic strategies for automated data analysis in large numbers of donors are still lacking. Here, we report a computational pipeline, called FlowGM, which minimizes operator input, is insensitive to compensation settings, and can be adapted to different analytic panels. A Gaussian Mixture Model (GMM)-based approach was utilized for initial clustering, with the number of clusters determined using Bayesian Information Criterion. Meta-clustering in a reference donor permitted automated identification of 24 cell types across four panels. Cluster labels were integrated into FCS files, thus permitting comparisons to manual gating. Cell numbers and coefficient of variation (CV) were similar between FlowGM and conventional gating for lymphocyte populations, but notably FlowGM provided improved discrimination of "hard-to-gate" monocyte and dendritic cell (DC) subsets. FlowGM thus provides rapid high-dimensional analysis of cell phenotypes and is amenable to cohort studies. Copyright © 2015. Published by Elsevier Inc.

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

Flow cytometric DNA ploidy analysis of ovarian granulosa cell tumors

NARCIS (Netherlands)

D. Chadha; C.J. Cornelisse; A. Schabert (A.)

1990-01-01

textabstractAbstract The nuclear DNA content of 50 ovarian tumors initially diagnosed as granulosa cell tumors was measured by flow cytometry using paraffin-embedded archival material. The follow-up period of the patients ranged from 4 months to 19 years. Thirty-eight tumors were diploid or

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.

Cooperative Strategies for Maximum-Flow Problem in Uncertain Decentralized Systems Using Reliability Analysis

Directory of Open Access Journals (Sweden)

Hadi Heidari Gharehbolagh

2016-01-01

Full Text Available This study investigates a multiowner maximum-flow network problem, which suffers from risky events. Uncertain conditions effect on proper estimation and ignoring them may mislead decision makers by overestimation. A key question is how self-governing owners in the network can cooperate with each other to maintain a reliable flow. Hence, the question is answered by providing a mathematical programming model based on applying the triangular reliability function in the decentralized networks. The proposed method concentrates on multiowner networks which suffer from risky time, cost, and capacity parameters for each network’s arcs. Some cooperative game methods such as τ-value, Shapley, and core center are presented to fairly distribute extra profit of cooperation. A numerical example including sensitivity analysis and the results of comparisons are presented. Indeed, the proposed method provides more reality in decision-making for risky systems, hence leading to significant profits in terms of real cost estimation when compared with unforeseen effects.

Application of analytical procedure on system reliability, GO-FLOW

International Nuclear Information System (INIS)

Matsuoka, Takeshi; Fukuto, Junji; Mitomo, Nobuo; Miyazaki, Keiko; Matsukura, Hiroshi; Kobayashi, Michiyuki

2000-01-01

In the Ship Research Institute, research and development of GO-FLOW procedure with various advanced functions as a system reliability analysis method occupying main part of the probabilistic safety assessment (PSA) were promoted. In this study, as an important evaluation technique on executing PSA with lower than level 3, by intending fundamental upgrading of the GO-FLOW procedure, a safety assessment system using the GO-FLOW as well as an analytical function coupling of dynamic behavior analytical function and physical behavior of the system with stochastic phenomenon change were developed. In 1998 fiscal year, preparation and verification of various functions such as dependence addition between the headings, rearrangement in order of time, positioning of same heading to plural positions, calculation of forming frequency with elapsing time were carried out. And, on a simulation analysis function of accident sequence, confirmation on analysis covering all of main accident sequence in the reactor for improved marine reactor, MRX was carried out. In addition, a function near automatically producible on input data for analysis was also prepared. As a result, the conventional analysis not always easy understanding on analytical results except an expert of PSA was solved, and understanding of the accident phenomenon, verification of validity on analysis, feedback to analysis, and feedback to design could be easily carried out. (G.K.)

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...

Use of flow cytometry for the possible identification of radio-induced changes in DNA of animal cells

International Nuclear Information System (INIS)

Spano, M.; Leonardi, M.; Cordelli, E.

1991-01-01

Since DNA is the main cellular target of ionizing irradiation, methods fit for analyzing DNA alterations should be able to discern irradiated versus control cells. Flow cytometry allows the rapid measurement of DNA content of single chromosomes or cell nuclei at very high resolution on a statistically significant sample. Alterations of chromatine structure can also be analyzed by flow cytometry. Briefly, evaluation of in situ DNA resistance to denaturation can be evaluated by flow cytometric analysis of different staining pattern of single versus double strange regions of DNA. In the present work both approaches were used with the aim to recognize cells derived from an irradiated sample of breast chicken. Although flow cytometry has been demonstrated to be a useful tool to detect DNA alterations and has been widely used to detect damages on DNA induced by several physical and chemical agents, it was unable to detect clastogenic effects induced by electrons on DNA of chicken breast cells. Heavily irradiated nuclei, even if challenged by denaturating treatments that partially collapse chromatine organization, do not present differences from non irradiated samples after flow cytometric DNA content measurement. (16 refs)

Study on reliability analysis based on multilevel flow models and fault tree method

International Nuclear Information System (INIS)

Chen Qiang; Yang Ming

2014-01-01

Multilevel flow models (MFM) and fault tree method describe the system knowledge in different forms, so the two methods express an equivalent logic of the system reliability under the same boundary conditions and assumptions. Based on this and combined with the characteristics of MFM, a method mapping MFM to fault tree was put forward, thus providing a way to establish fault tree rapidly and realizing qualitative reliability analysis based on MFM. Taking the safety injection system of pressurized water reactor nuclear power plant as an example, its MFM was established and its reliability was analyzed qualitatively. The analysis result shows that the logic of mapping MFM to fault tree is correct. The MFM is easily understood, created and modified. Compared with the traditional fault tree analysis, the workload is greatly reduced and the modeling time is saved. (authors)

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.

Miniaturized flow cytometer with 3D hydrodynamic particle focusing and integrated optical elements applying silicon photodiodes

NARCIS (Netherlands)

Rosenauer, M.; Buchegger, W.; Finoulst, I.; Verhaert, P.D.E.M.; Vellekoop, M.

2010-01-01

In this study, the design, realization and measurement results of a novel optofluidic system capable of performing absorbance-based flow cytometric analysis is presented. This miniaturized laboratory platform, fabricated using SU-8 on a silicon substrate, comprises integrated polymer-based

Measurement of separase proteolytic activity in single living cells by a fluorogenic flow cytometry assay.

Directory of Open Access Journals (Sweden)

Wiltrud Haaß

Full Text Available ESPL1/Separase, an endopeptidase, is required for centrosome duplication and separation of sister-chromatides in anaphase of mitosis. Overexpression and deregulated proteolytic activity of Separase as frequently observed in human cancers is associated with the occurrence of supernumerary centrosomes, chromosomal missegregation and aneuploidy. Recently, we have hypothesized that increased Separase proteolytic activity in a small subpopulation of tumor cells may serve as driver of tumor heterogeneity and clonal evolution in chronic myeloid leukemia (CML. Currently, there is no quantitative assay to measure Separase activity levels in single cells. Therefore, we have designed a flow cytometry-based assay that utilizes a Cy5- and rhodamine 110 (Rh110-biconjugated Rad21 cleavage site peptide ([Cy5-D-R-E-I-M-R]2-Rh110 as smart probe and intracellular substrate for detection of Separase enzyme activity in living cells. As measured by Cy5 fluorescence the cellular uptake of the fluorogenic peptide was fast and reached saturation after 210 min of incubation in human histiocytic lymphoma U937 cells. Separase activity was recorded as the intensity of Rh110 fluorescence released after intracellular peptide cleavage providing a linear signal gain within a 90-180 min time slot. Compared to conventional cell extract-based methods the flow cytometric assay delivers equivalent results but is more reliable, bypasses the problem of vague loading controls and unspecific proteolysis associated with whole cell extracts. Especially suited for the investigaton of blood- and bone marrow-derived hematopoietic cells the flow cytometric Separase assay allows generation of Separase activity profiles that tell about the number of Separase positive cells within a sample i.e. cells that currently progress through mitosis and about the range of intercellular variation in Separase activity levels within a cell population. The assay was used to quantify Separase proteolytic

Detection and quantification of live, apoptotic, and necrotic human peripheral lymphocytes by single-laser flow cytometry.

Science.gov (United States)

Liegler, T J; Hyun, W; Yen, T S; Stites, D P

1995-05-01

Regulation of peripheral lymphocyte number involves a poorly understood balance between cell renewal and loss. Disrupting this balance leads to a large number of disease states. Methods which allow qualitative and quantitative measurements of cell viability are increasingly valuable to studies directed at revealing the mechanisms underlying apoptotic and necrotic cell death. Here, we have characterized a method using single-laser flow cytometry that differentiates and quantifies the relative number of live, apoptotic, and late-stage apoptotic and necrotic peripheral lymphocytes. Following in vitro gamma irradiation and staining with acridine orange in combination with ethidium bromide, three distinct populations were seen by bivariate analysis of green versus red fluorescence. The identity of each distinct fluorescent population (whether live, apoptotic, or necrotic) was determined by sorting and examination of cellular morphology by electron microscopy. This flow cytometric method is directly compared with the techniques of trypan blue exclusion and DNA fragmentation to quantify cell death following exposure to various doses of in vitro gamma irradiation and postirradiation incubation times. We extend our findings to illustrate the utility of this method beyond analyzing radiation-induced apoptotic peripheral blood mononuclear cells (PBMC); similar fluorescent patterns are shown for radiation- and corticosteroid-treated murine thymocytes, activated human PBMC, and PBMC from human immunodeficiency virus-infected individuals. Our results demonstrate that dual-parameter flow cytometric analysis of acridine orange-ethidium bromide-stained lymphocytes is overall a superior method with increased sensitivity, greater accuracy, and decreased subjectivity in comparison with the other methods tested. By using standard laser and filter settings commonly available to flow cytometric laboratories, this method allows rapid measurement of a large number of cells from a

A new cut-based algorithm for the multi-state flow network reliability problem

International Nuclear Information System (INIS)

Yeh, Wei-Chang; Bae, Changseok; Huang, Chia-Ling

2015-01-01

Many real-world systems can be modeled as multi-state network systems in which reliability can be derived in terms of the lower bound points of level d, called d-minimal cuts (d-MCs). This study proposes a new method to find and verify obtained d-MCs with simple and useful found properties for the multi-state flow network reliability problem. The proposed algorithm runs in O(mσp) time, which represents a significant improvement over the previous O(mp 2 σ) time bound based on max-flow/min-cut, where p, σ and m denote the number of MCs, d-MC candidates and edges, respectively. The proposed algorithm also conquers the weakness of some existing methods, which failed to remove duplicate d-MCs in special cases. A step-by-step example is given to demonstrate how the proposed algorithm locates and verifies all d-MC candidates. As evidence of the utility of the proposed approach, we present extensive computational results on 20 benchmark networks in another example. The computational results compare favorably with a previously developed algorithm in the literature. - Highlights: • A new method is proposed to find all d-MCs for the multi-state flow networks. • The proposed method can prevent the generation of d-MC duplicates. • The proposed method is simpler and more efficient than the best-known algorithms

Solid KHT tumor dispersal for flow cytometric cell kinetic analysis

International Nuclear Information System (INIS)

Pallavicini, M.G.; Folstad, L.J.; Dunbar, C.

1981-01-01

A bacterial neutral protease was used to disperse KHT solid tumors into single cell suspensions suitable for routine cell kinetic analysis by flow cytometry and for clonogenic cell survival. Neutral protease disaggregation under conditions which would be suitable for routine tumor dispersal was compared with a trypsin/DNase procedure. Cell yield, clonogenic cell survival, DNA distributions of untreated and drug-perturbed tumors, rates of radioactive precursor incorporation during the cell cycle, and preferential cell cycle phase-specific cell loss were investigated. Tumors dispersed with neutral protease yielded approximately four times more cells than those dispersed with trypsin/DNase and approximately a 1.5-fold higher plating efficiency in a semisolid agar system. Quantitative analysis of DNA distributions obtained from untreated and cytosine-arabinoside-perturbed tumors produced similar results with both dispersal procedures. The rates of incorporation of tritiated thymidine during the cell cycle were also similar with neutral protease and trypsin/DNase dispersal. Preferential phase-specific cell loss was not obseved with either technique. We find that neutral protease provides good single cell suspensions of the KHT tumor for cell survival measurements and for cell kinetic analysis of drug-induced perturbations by flow cytometry. In addition, the high cell yields facilitate electronic cell sorting where large numbers of cells are often required

Limits on reliable information flows through stochastic populations.

Science.gov (United States)

Boczkowski, Lucas; Natale, Emanuele; Feinerman, Ofer; Korman, Amos

2018-06-06

Biological systems can share and collectively process information to yield emergent effects, despite inherent noise in communication. While man-made systems often employ intricate structural solutions to overcome noise, the structure of many biological systems is more amorphous. It is not well understood how communication noise may affect the computational repertoire of such groups. To approach this question we consider the basic collective task of rumor spreading, in which information from few knowledgeable sources must reliably flow into the rest of the population. We study the effect of communication noise on the ability of groups that lack stable structures to efficiently solve this task. We present an impossibility result which strongly restricts reliable rumor spreading in such groups. Namely, we prove that, in the presence of even moderate levels of noise that affect all facets of the communication, no scheme can significantly outperform the trivial one in which agents have to wait until directly interacting with the sources-a process which requires linear time in the population size. Our results imply that in order to achieve efficient rumor spread a system must exhibit either some degree of structural stability or, alternatively, some facet of the communication which is immune to noise. We then corroborate this claim by providing new analyses of experimental data regarding recruitment in Cataglyphis niger desert ants. Finally, in light of our theoretical results, we discuss strategies to overcome noise in other biological systems.

Capacity Expansion and Reliability Evaluation on the Networks Flows with Continuous Stochastic Functional Capacity

Directory of Open Access Journals (Sweden)

F. Hamzezadeh

2014-01-01

Full Text Available In many systems such as computer network, fuel distribution, and transportation system, it is necessary to change the capacity of some arcs in order to increase maximum flow value from source s to sink t, while the capacity change incurs minimum cost. In real-time networks, some factors cause loss of arc’s flow. For example, in some flow distribution systems, evaporation, erosion or sediment in pipes waste the flow. Here we define a real capacity, or the so-called functional capacity, which is the operational capacity of an arc. In other words, the functional capacity of an arc equals the possible maximum flow that may pass through the arc. Increasing the functional arcs capacities incurs some cost. There is a certain resource available to cover the costs. First, we construct a mathematical model to minimize the total cost of expanding the functional capacities to the required levels. Then, we consider the loss of flow on each arc as a stochastic variable and compute the system reliability.

Designing reliability information flows

International Nuclear Information System (INIS)

Petkova, Valia T.; Lu Yuan; Ion, Roxana A.; Sander, Peter C.

2005-01-01

It is well-known [Reliab. Eng. Syst. Saf. 75 (2002) 295] that in modern development processes it is essential to have an information flow structure that facilitates fast feedback from product users (customers) to departments at the front end, in particular development and production. As information is only relevant if it is used when taking decisions, this paper presents a guideline for building field feedback information flows that facilitate the decision taking during the product creation and realisation process. The guideline takes into consideration that the type of decisions depends on the span-of-control, therefore following Parsons [Structure and Process in Modern Societies (1990)] the span-of-control is subdivided into the following three levels: strategic, tactic, and executive. The guideline is illustrated with a case in which it is used for analysing the quality of existing field feedback flows

Research on application of technique for analyzing system reliability, GO-FLOW

International Nuclear Information System (INIS)

Matsuoka, Takeshi; Fukuto, Junji; Sugasawa, Shinobu; Mitomo, Nobuo; Miyazaki, Keiko; Hirao, Yoshihiro; Kobayashi, Michiyuki

1997-01-01

As the method of evaluation, probabilistic safety assessment (PSA) has been introduced in nuclear power field, and began to play important role in plant design and safety examination. In the Ship Research Institute, as the technique for analyzing system reliability which takes the main part of PSA, the research on developing the GO-FLOW technique which has various advanced functions has been carried out. In this research, the functions of the GO-FLOW technique are improved, and the function of the dynamic behavior analysis for systems and the analysis function for the combination of the physical behavior of systems and the change of probabilistic events are developed, further, the function of extracting main accident sequence by utilizing the GO-FLOW technique is prepared. As for the analysis of dynamic behavior, the sample problem on hold-up tank was investigated. As to the extraction of main accident sequence, the fundamental part of the function of event tree analysis was consolidated, and the function of setting branching probability was given. As to the indication of plant behavior, the simulator for improved marine reactor MRX was developed. (K.I.)

Measurement of Soluble Biomarkers by Flow Cytometry

OpenAIRE

Antal-Szalm?s, P?ter; Nagy, B?la; Debreceni, Ildik? Beke; Kappelmayer, J?nos

2013-01-01

Microparticle based flow cytometric assays for determination of the level of soluble biomarkers are widely used in several research applications and in some diagnostic setups. The major advantages of these multiplex systems are that they can measure a large number of analytes (up to 500) at the same time reducing assay time, costs and sample volume. Most of these assays are based on antigen-antibody interactions and work as traditional immunoassays, but nucleic acid alterations ? by using spe...

Lipid nanoparticles assessment by flow cytometry.

Science.gov (United States)

Bryła, Anna; Juzwa, Wojciech; Weiss, Marek; Lewandowicz, Grażyna

2017-03-30

Liposomes are promising carriers for drugs and bioactive compounds. Size and structure are their crucial parameters. Thus, it is essential to assess individual vesicles as prepared. Currently available techniques fail to measure liposome's size and structure simultaneously, with a high throughput. To solve this problem, we have developed a novel, flow cytometric method quantifying liposomes. Firstly, the following fluorescent staining combinations were tested: DiD/TO, Rh123/DiD, Syto9/DiD. Further, chosen fluorochromes were used to compare three populations of vesicles: raw (R), obtained by thin film hydration and extruded ones (populations E10 and E21). Dynamic light scattering (DLS) was used for determination of average diameter and size distribution of nanocarriers. Structural differences between the raw and the extruded liposomes, as well as additional information concerning vesicles size were acquired employing atomic force microscopy (AFM). DLS analysis indicated that, three distinct populations of vesicles were obtained. Liposomes were characterized by mean diameter of 323nm, 220nm and 170nm for population R, E10 and E21 respectively. All the populations were stable and revealed zeta potential of -29mV. AFM confirmed that raw and extruded liposomes were differed in structure. DiD/TO was the optimal fluorochrome combination that enabled to resolve distinctly the sub-populations of liposomes. Results obtained by flow cytometry were in a good agreement with those from DLS and AFM. It was proved that, flow cytometry, when proper fluorescent dyes are used, is an adequate method for liposomes assessment. The proposed method enables fast and reliable analysis of liposomes in their native environment. Copyright © 2017 Elsevier B.V. All rights reserved.

Using minimal cuts to evaluate the system reliability of a stochastic-flow network with failures at nodes and arcs

International Nuclear Information System (INIS)

Lin, Y.-K.

2002-01-01

This paper deals with a stochastic-flow network in which each node and arc has a designated capacity, which will have different lower levels due to various partial and complete failures. We try to evaluate the system reliability that the maximum flow of the network is not less than a demand (d+1). A simple algorithm in terms of minimal cuts is first proposed to generate all upper boundary points for d, and then the system reliability can be calculated in terms of such points. The upper boundary point for d is a maximal vector, which represents the capacity of each component (arc or node), such that the maximum flow of the network is d. A computer example is shown to illustrate the solution procedure

Absolute counting of neutrophils in whole blood using flow cytometry.

Science.gov (United States)

Brunck, Marion E G; Andersen, Stacey B; Timmins, Nicholas E; Osborne, Geoffrey W; Nielsen, Lars K

2014-12-01

Absolute neutrophil count (ANC) is used clinically to monitor physiological dysfunctions such as myelosuppression or infection. In the research laboratory, ANC is a valuable measure to monitor the evolution of a wide range of disease states in disease models. Flow cytometry (FCM) is a fast, widely used approach to confidently identify thousands of cells within minutes. FCM can be optimised for absolute counting using spiked-in beads or by measuring the sample volume analysed. Here we combine the 1A8 antibody, specific for the mouse granulocyte protein Ly6G, with flow cytometric counting in straightforward FCM assays for mouse ANC, easily implementable in the research laboratory. Volumetric and Trucount™ bead assays were optimized for mouse neutrophils, and ANC values obtained with these protocols were compared to ANC measured by a dual-platform assay using the Orphee Mythic 18 veterinary haematology analyser. The single platform assays were more precise with decreased intra-assay variability compared with ANC obtained using the dual protocol. Defining ANC based on Ly6G expression produces a 15% higher estimate than the dual protocol. Allowing for this difference in ANC definition, the flow cytometry counting assays using Ly6G can be used reliably in the research laboratory to quantify mouse ANC from a small volume of blood. We demonstrate the utility of the volumetric protocol in a time-course study of chemotherapy induced neutropenia using four drug regimens. © 2014 International Society for Advancement of Cytometry.

Coexpression of multidrug resistance involve proteins: a flow cytometric analysis.

Science.gov (United States)

Boutonnat, J; Bonnefoix, T; Mousseau, M; Seigneurin, D; Ronot, X

1998-01-01

Cross resistance to multiple natural cytotoxic products represents a major obstacle in myeloblastic acute leukaemia (AML). Multidrug resistance (MDR) often involves overexpression of plasma membrane drug transporter P-glycoprotein (PGP) or the resistance associated protein (MRP). Recently, a protein overexpressed in a non-PGP MDR lung cancer cell line and termed lung resistance related protein (LRP) was identified. These proteins are known to be associated with a bad prognosis in AML. We have developed a triple indirect labelling analysed by flow cytometry to detect the coexpression of these proteins. Since no cell line expressing all three antigens is known, we mixed K562 cells (resistant to Adriblastine, PGP+, MRP-, LRP-) with GLC4 cells (resistant to Adriblastine, PGP-, MRP+, LRP+) to create a model system to test the method. The antibodies used were UIC2 for PGP, MRPm6 for MRP and LRP56 for LRP. They were revealed by Fab'2 coupled with Fluoresceine-isothiocyanate, Phycoerythrin or Tricolor with isotype specificity. Cells were fixed and permeabilized after PGP labelling because MRPm6 and LRP56 recognize intracellular epitopes. PGP and LRP were easily detected. MRP is expressed at relatively low levels and was more difficult to detect because in the triple labelling the non specific staining was higher than in a single labelling. Despite the increased background in the triple labelling we were able to detect coexpression of PGP, MRP, LRP by flow cytometry. This method appears to be very useful to detect coexpression of markers in AML. Such coexpression could modify the therapeutic approach with revertants.

Suncor maintenance and reliability

Energy Technology Data Exchange (ETDEWEB)

Little, S. [Suncor Energy, Calgary, AB (Canada)

2006-07-01

Fleet maintenance and reliability at Suncor Energy was discussed in this presentation, with reference to Suncor Energy's primary and support equipment fleets. This paper also discussed Suncor Energy's maintenance and reliability standard involving people, processes and technology. An organizational maturity chart that graphed organizational learning against organizational performance was illustrated. The presentation also reviewed the maintenance and reliability framework; maintenance reliability model; the process overview of the maintenance and reliability standard; a process flow chart of maintenance strategies and programs; and an asset reliability improvement process flow chart. An example of an improvement initiative was included, with reference to a shovel reliability review; a dipper trip reliability investigation; bucket related failures by type and frequency; root cause analysis of the reliability process; and additional actions taken. Last, the presentation provided a graph of the results of the improvement initiative and presented the key lessons learned. tabs., figs.

Detection of bacteriophage-infected cells of Lactococcus lactis using flow cytometry

DEFF Research Database (Denmark)

Michelsen, Ole; Cuesta-Dominguez, Álvaro; Albrektsen, Bjarne

2007-01-01

Bacteriophage infection in dairy fermentation constitutes a serious problem worldwide. We have studied bacteriophage infection in Lactococcus lactis by using the flow cytometer. The first effect of the infection of the bacterium is a change from cells in chains toward single cells. We interpret...... describe a new method for detection of phage infection in Lactococcus lactis dairy cultures. The method is based on flow cytometric detection of cells with low-density cell walls. The method allows fast and early detection of phage-infected bacteria, independently of which phage has infected the culture...

Advances in Automated Plankton Imaging: Enhanced Throughput, Automated Staining, and Extended Deployment Modes for Imaging FlowCytobot

Science.gov (United States)

Sosik, H. M.; Olson, R. J.; Brownlee, E.; Brosnahan, M.; Crockford, E. T.; Peacock, E.; Shalapyonok, A.

2016-12-01

Imaging FlowCytobot (IFCB) was developed to fill a need for automated identification and monitoring of nano- and microplankton, especially phytoplankton in the size range 10 200 micrometer, which are important in coastal blooms (including harmful algal blooms). IFCB uses a combination of flow cytometric and video technology to capture high resolution (1 micrometer) images of suspended particles. This proven, now commercially available, submersible instrument technology has been deployed in fixed time series locations for extended periods (months to years) and in shipboard laboratories where underway water is automatically analyzed during surveys. Building from these successes, we have now constructed and evaluated three new prototype IFCB designs that extend measurement and deployment capabilities. To improve cell counting statistics without degrading image quality, a high throughput version (IFCB-HT) incorporates in-flow acoustic focusing to non-disruptively pre-concentrate cells before the measurement area of the flow cell. To extend imaging to all heterotrophic cells (even those that do not exhibit chlorophyll fluorescence), Staining IFCB (IFCB-S) incorporates automated addition of a live-cell fluorescent stain (fluorescein diacetate) to samples before analysis. A horizontally-oriented IFCB-AV design addresses the need for spatial surveying from surface autonomous vehicles, including design features that reliably eliminate air bubbles and mitigate wave motion impacts. Laboratory evaluation and test deployments in waters near Woods Hole show the efficacy of each of these enhanced IFCB designs.

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

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

Image Cytometric Analysis of Algal Spores for Evaluation of Antifouling Activities of Biocidal Agents.

Science.gov (United States)

Il Koo, Bon; Lee, Yun-Soo; Seo, Mintae; Seok Choi, Hyung; Leng Seah, Geok; Nam, Taegu; Nam, Yoon Sung

2017-07-31

Chemical biocides have been widely used as marine antifouling agents, but their environmental toxicity impose regulatory restriction on their use. Although various surrogate antifouling biocides have been introduced, their comparative effectiveness has not been well investigated partly due to the difficulty of quantitative evaluation of their antifouling activity. Here we report an image cytometric method to quantitatively analyze the antifouling activities of seven commercial biocides using Ulva prolifera as a target organism, which is known to be a dominant marine species causing soft fouling. The number of spores settled on a substrate is determined through image analysis using the intrinsic fluorescence of chlorophylls in the spores. Pre-determined sets of size and shape of spores allow for the precise determination of the number of settled spores. The effects of biocide concentration and combination of different biocides on the spore settlement are examined. No significant morphological changes of Ulva spores are observed, but the amount of adhesive pad materials is appreciably decreased in the presence of biocides. It is revealed that the growth rate of Ulva is not directly correlated with the antifouling activities against the settlement of Ulva spores. This work suggests that image cytometric analysis is a very convenient, fast-processable method to directly analyze the antifouling effects of biocides and coating materials.

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

Decision Diagram Based Symbolic Algorithm for Evaluating the Reliability of a Multistate Flow Network

Directory of Open Access Journals (Sweden)

Rongsheng Dong

2016-01-01

Full Text Available Evaluating the reliability of Multistate Flow Network (MFN is an NP-hard problem. Ordered binary decision diagram (OBDD or variants thereof, such as multivalued decision diagram (MDD, are compact and efficient data structures suitable for dealing with large-scale problems. Two symbolic algorithms for evaluating the reliability of MFN, MFN_OBDD and MFN_MDD, are proposed in this paper. In the algorithms, several operating functions are defined to prune the generated decision diagrams. Thereby the state space of capacity combinations is further compressed and the operational complexity of the decision diagrams is further reduced. Meanwhile, the related theoretical proofs and complexity analysis are carried out. Experimental results show the following: (1 compared to the existing decomposition algorithm, the proposed algorithms take less memory space and fewer loops. (2 The number of nodes and the number of variables of MDD generated in MFN_MDD algorithm are much smaller than those of OBDD built in the MFN_OBDD algorithm. (3 In two cases with the same number of arcs, the proposed algorithms are more suitable for calculating the reliability of sparse networks.

Assessment of system reliability for a stochastic-flow distribution network with the spoilage property

Science.gov (United States)

Lin, Yi-Kuei; Huang, Cheng-Fu; Yeh, Cheng-Ta

2016-04-01

In supply chain management, satisfying customer demand is the most concerned for the manager. However, the goods may rot or be spoilt during delivery owing to natural disasters, inclement weather, traffic accidents, collisions, and so on, such that the intact goods may not meet market demand. This paper concentrates on a stochastic-flow distribution network (SFDN), in which a node denotes a supplier, a transfer station, or a market, while a route denotes a carrier providing the delivery service for a pair of nodes. The available capacity of the carrier is stochastic because the capacity may be partially reserved by other customers. The addressed problem is to evaluate the system reliability, the probability that the SFDN can satisfy the market demand with the spoilage rate under the budget constraint from multiple suppliers to the customer. An algorithm is developed in terms of minimal paths to evaluate the system reliability along with a numerical example to illustrate the solution procedure. A practical case of fruit distribution is presented accordingly to emphasise the management implication of the system reliability.

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

KAUST Repository

Prest, Emmanuelle I E C; Hammes, Frederik A.; Kö tzsch, Stefan; van Loosdrecht, Mark C.M.; Vrouwenvelder, Johannes S.

2013-01-01

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.

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.

Rapid assay for cell age response to radiation by electronic volume flow cell sorting

International Nuclear Information System (INIS)

Freyer, J.P.; Wilder, M.E.; Raju, M.R.

1987-01-01

A new technique is described for measuring cell survival as a function of cell cycle position using flow cytometric cell sorting on the basis of electronic volume signals. Sorting of cells into different cell age compartments is demonstrated for three different cell lines commonly used in radiobiological research. Using flow cytometric DNA content analysis and [ 3 H]thymidine autoradiography of the sorted cell populations, it is demonstrated that resolution of the age compartment separation is as good as or better than that reported for other cell synchronizing techniques. Variation in cell survival as a function of position in the cell cycle after a single dose of radiation as measured by volume cell sorting is similar to that determined by other cell synchrony techniques. Advantages of this method include: (1) no treatment of the cells is required, thus, this method is noncytotoxic; (2) no cell cycle progression is needed to obtain different cell age compartments; (3) the cell population can be held in complete growth medium at any desired temperature during sorting; (4) a complete radiation age - response assay can be plated in 2 h. Applications of this method are discussed, along with some technical limitations. (author)

Standardization of 8-color flow cytometry across different flow cytometer instruments: A feasibility study in clinical laboratories in Switzerland.

Science.gov (United States)

Glier, Hana; Heijnen, Ingmar; Hauwel, Mathieu; Dirks, Jan; Quarroz, Stéphane; Lehmann, Thomas; Rovo, Alicia; Arn, Kornelius; Matthes, Thomas; Hogan, Cassandra; Keller, Peter; Dudkiewicz, Ewa; Stüssi, Georg; Fernandez, Paula

2017-07-29

The EuroFlow Consortium developed a fully standardized flow cytometric approach from instrument settings, through antibody panel, reagents and sample preparation protocols, to data acquisition and analysis. The Swiss Cytometry Society (SCS) promoted a study to evaluate the feasibility of using such standardized measurements of 8-color data across two different flow cytometry platforms - Becton Dickinson (BD) FACSCanto II and Beckman Coulter (BC) Navios, aiming at increasing reproducibility and inter-laboratory comparability of immunophenotypic data in clinical laboratories in Switzerland. The study was performed in two phases, i.e. a learning phase (round 1) and an analytical phase (rounds 2 and 3) consisting of a total of three rounds. Overall, 10 laboratories using BD FACSCanto II (n=6) or BC Navios (n=4) flow cytometers participated. Each laboratory measured peripheral blood samples from healthy donors stained with a uniform antibody panel of reagents - EuroFlow Lymphoid Screening Tube (LST) - applying the EuroFlow standardized protocols for instrument setup and sample preparation (www.EuroFlow.org). All data files were analyzed centrally and median fluorescence intensity (MedFI) values for individual markers on defined lymphocyte subsets were recorded; variability from reference MedFI values was assessed using performance scores. Data troubleshooting and discussion of the results with the participants followed after each round at SCS meetings. The results of the learning phase demonstrated that standardized instrument setup and data acquisition are feasible in routine clinical laboratories without previous experience with EuroFlow. During the analytical phase, highly comparable data were obtained at the different laboratories using either BD FACSCanto II or BC Navios. The coefficient of variation of MedFI for 7 of 11 markers performed repeatedly below 30%. In the last study round, 89% of participants scored over 90% MedFI values within the acceptance criteria

Genome size variation in the pine fusiform rust pathogen Cronartium quercuum f.sp. fusiforme as determined by flow cytometry

Science.gov (United States)

Claire L Anderson; Thomas L Kubisiak; C Dana Nelson; Jason A Smith; John M Davis

2010-01-01

The genome size of the pine fusiform rust pathogen Cronartium quercuum f.sp. fusiforme (Cqf) was determined by flow cytometric analysis of propidium iodide-stained, intact haploid pycniospores with haploid spores of two genetically well characterized fungal species, Sclerotinia sclerotiorum and Puccinia graminis f.sp. tritici, as size standards. The Cqf haploid genome...

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.

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.

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

KAUST Repository

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

2017-01-01

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.

A simple MC-based algorithm for evaluating reliability of stochastic-flow network with unreliable nodes

International Nuclear Information System (INIS)

Yeh, W.-C.

2004-01-01

A MP/minimal cutset (MC) is a path/cut set such that if any edge is removed from this path/cut set, then the remaining set is no longer a path/cut set. An intuitive method is proposed to evaluate the reliability in terms of MCs in a stochastic-flow network subject to both edge and node failures under the condition that all of the MCs are given in advance. This is an extension of the best of known algorithms for solving the d-MC (a special MC but formatted in a system-state vector, where d is the lower bound points of the system capacity level) problem from the stochastic-flow network without unreliable nodes to with unreliable nodes by introducing some simple concepts. These concepts were first developed in the literature to implement the proposed algorithm to reduce the number of d-MC candidates. This method is more efficient than the best of known existing algorithms regardless if the network has or does not have unreliable nodes. Two examples are illustrated to show how the reliability is determined using the proposed algorithm in the network with or without unreliable nodes. The computational complexity of the proposed algorithm is analyzed and compared with the existing methods

Overview of system reliability analyses for PSA

International Nuclear Information System (INIS)

Matsuoka, Takeshi

2012-01-01

Overall explanations are given for many matters relating to system reliability analysis. Systems engineering, Operations research, Industrial engineering, Quality control are briefly explained. Many system reliability analysis methods including advanced methods are introduced. Discussions are given for FMEA, reliability block diagram, Markov model, Petri net, Bayesian network, goal tree success tree, dynamic flow graph methodology, cell-to-cell mapping technique, the GO-FLOW and others. (author)

Roads at risk - the impact of debris flows on road network reliability and vulnerability in southern Norway

Science.gov (United States)

Meyer, Nele Kristin; Schwanghart, Wolfgang; Korup, Oliver

2014-05-01

Norwegian's road network is frequently affected by debris flows. Both damage repair and traffic interruption generate high economic losses and necessitate a rigorous assessment of where losses are expected to be high and where preventive measures should be focused on. In recent studies, we have developed susceptibility and trigger probability maps that serve as input into a hazard calculation at the scale of first-order watersheds. Here we combine these results with graph theory to assess the impact of debris flows on the road network of southern Norway. Susceptibility and trigger probability are aggregated for individual road sections to form a reliability index that relates to the failure probability of a link that connects two network vertices, e.g., road junctions. We define link vulnerability as a function of traffic volume and additional link failure distance. Additional link failure distance is the extra length of the alternative path connecting the two associated link vertices in case the network link fails and is calculated by a shortest-path algorithm. The product of network reliability and vulnerability indices represent the risk index. High risk indices identify critical links for the Norwegian road network and are investigated in more detail. Scenarios demonstrating the impact of single or multiple debris flow events are run for the most important routes between seven large cities in southern Norway. First results show that the reliability of the road network is lowest in the central and north-western part of the study area. Road network vulnerability is highest in the mountainous regions in central southern Norway where the road density is low and in the vicinity of cities where the traffic volume is large. The scenarios indicate that city connections that have their shortest path via routes crossing the central part of the study area have the highest risk of route failure.

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...

What Is Fanconi Anemia?

Science.gov (United States)

... people who don't have FA. Cytometric Flow Analysis Cytometric flow analysis, or CFA, is done in a lab. This ... to know about resources that can give you emotional support and helpful information about FA and its ...

Reliable Design Versus Trust

Science.gov (United States)

Berg, Melanie; LaBel, Kenneth A.

2016-01-01

This presentation focuses on reliability and trust for the users portion of the FPGA design flow. It is assumed that the manufacturer prior to hand-off to the user tests FPGA internal components. The objective is to present the challenges of creating reliable and trusted designs. The following will be addressed: What makes a design vulnerable to functional flaws (reliability) or attackers (trust)? What are the challenges for verifying a reliable design versus a trusted design?

Analysis of UV-excited fluorochromes by flow cytometry using near-ultraviolet laser diodes.

Science.gov (United States)

Telford, William G

2004-09-01

Violet laser diodes have become common and reliable laser sources for benchtop flow cytometers. While these lasers are very useful for a variety of violet and some ultraviolet-excited fluorochromes (e.g., DAPI), they do not efficiently excite most UV-stimulated probes. In this study, the next generation of InGaN near-UV laser diodes (NUVLDs) emitting in the 370-375-nm range have been evaluated as laser sources for cuvette-based flow cytometers. Several NUVLDs, ranging in wavelength from 370 to 374 nm and in power level from 1.5 to 10 mW, were mounted on a BD Biosciences LSR II and evaluated for their ability to excite cells labeled with the UV DNA binding dye DAPI, several UV phenotyping fluorochromes (including Alexa Fluor 350, Marina Blue, and quantum dots), and the fluorescent calcium chelator indo-1. NUVLDs at the 8-10-mW power range gave detection sensitivity levels comparable to more powerful solid-state and ion laser sources, using low-fluorescence microsphere beads as measurement standards. NUVLDs at all tested power levels allowed extremely high-resolution DAPI cell cycle analysis, and sources in the 8-10-mW power range excited Alexa Fluor 350, Marina Blue, and a variety of quantum dots at virtually the same signal-to-noise ratios as more powerful UV sources. These evaluations indicate that near-UV laser diodes installed on a cuvette-based flow cytometer performed nearly as well as more powerful solid-state UV lasers on the same instrumentation, and comparably to more powerful ion lasers on a jet-in-air system, and. Despite their limited power, integration of these small and inexpensive lasers into benchtop flow cytometers should allow the use of flow cytometric applications requiring UV excitation on a wide variety of instruments. Copyright 2004 Wiley-Liss, Inc.

Characterization of glycosylphosphatidylinositol biosynthesis defects by clinical features, flow cytometry, and automated image analysis

DEFF Research Database (Denmark)

Knaus, Alexej; Pantel, Jean Tori; Pendziwiat, Manuela

2018-01-01

, the increasing number of individuals with a GPIBD shows that hyperphosphatasia is a variable feature that is not ideal for a clinical classification. METHODS: We studied the discriminatory power of multiple GPI-linked substrates that were assessed by flow cytometry in blood cells and fibroblasts of 39 and 14...... those with PIGA mutations. Although the impairment of GPI-linked substrates is supposed to play the key role in the pathophysiology of GPIBDs, we could not observe gene-specific profiles for flow cytometric markers or a correlation between their cell surface levels and the severity of the phenotype...

Determination of Multiphase Flow Meter Reliability and Development of Correction Charts for the Prediction of Oilfield Fluid Flow Rates

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Samuel S. MOFUNLEWI

2008-06-01

Full Text Available The aim of field testing of Multiphase Flow Meter (MPFM is to show whether its accuracy compares favourably with that of the Test Separator in accurately measuring the three production phases (oil, gas and water as well as determining meter reliability in field environment. This study evaluates field test results of the MPFM as compared to reference conventional test separators. Generally, results show that MPFM compares favourably with Test Separator within the specified range of accuracy.At the moment, there is no legislation for meter proving technique for MPFM. However, this study has developed calibration charts that can be used to correct and improve meter accuracy.

A hybrid load flow and event driven simulation approach to multi-state system reliability evaluation

International Nuclear Information System (INIS)

George-Williams, Hindolo; Patelli, Edoardo

2016-01-01

Structural complexity of systems, coupled with their multi-state characteristics, renders their reliability and availability evaluation difficult. Notwithstanding the emergence of various techniques dedicated to complex multi-state system analysis, simulation remains the only approach applicable to realistic systems. However, most simulation algorithms are either system specific or limited to simple systems since they require enumerating all possible system states, defining the cut-sets associated with each state and monitoring their occurrence. In addition to being extremely tedious for large complex systems, state enumeration and cut-set definition require a detailed understanding of the system's failure mechanism. In this paper, a simple and generally applicable simulation approach, enhanced for multi-state systems of any topology is presented. Here, each component is defined as a Semi-Markov stochastic process and via discrete-event simulation, the operation of the system is mimicked. The principles of flow conservation are invoked to determine flow across the system for every performance level change of its components using the interior-point algorithm. This eliminates the need for cut-set definition and overcomes the limitations of existing techniques. The methodology can also be exploited to account for effects of transmission efficiency and loading restrictions of components on system reliability and performance. The principles and algorithms developed are applied to two numerical examples to demonstrate their applicability. - Highlights: • A discrete event simulation model based on load flow principles. • Model does not require system path or cut sets. • Applicable to binary and multi-state systems of any topology. • Supports multiple output systems with competing demand. • Model is intuitive and generally applicable.

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

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 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 of dynamic safety system (DSS) shows that the estimated reliability of the system is quite reasonable and realistic. (author)

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.

Monodisperse Water-in-Oil-in-Water (W/O/W Double Emulsion Droplets as Uniform Compartments for High-Throughput Analysis via Flow Cytometry

Directory of Open Access Journals (Sweden)

Jing Yan

2013-12-01

Full Text Available Here we report the application of monodisperse double emulsion droplets, produced in a single step within partially hydrophilic/partially hydrophobic microfluidic devices, as defined containers for quantitative flow cytometric analysis. Samples with varying fluorophore concentrations were generated, and a clear correlation between dye concentration and fluorescence signals was observed.

Molecular Characterization of Gastric Epithelial Cells Using Flow Cytometry

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Kevin A. Bockerstett

2018-04-01

Full Text Available The ability to analyze individual epithelial cells in the gastric mucosa would provide important insight into gastric disease, including chronic gastritis and progression to gastric cancer. However, the successful isolation of viable gastric epithelial cells (parietal cells, neck cells, chief cells, and foveolar cells from gastric glands has been limited due to difficulties in tissue processing. Furthermore, analysis and interpretation of gastric epithelial cell flow cytometry data has been difficult due to the varying sizes and light scatter properties of the different epithelial cells, high levels of autofluorescence, and poor cell viability. These studies were designed to develop a reliable method for isolating viable single cells from the corpus of stomachs and to optimize analyses examining epithelial cells from healthy and diseased stomach tissue by flow cytometry. We performed a two stage enzymatic digestion in which collagenase released individual gastric glands from the stromal tissue of the corpus, followed by a Dispase II digestion that dispersed these glands into greater than 1 × 106 viable single cells per gastric corpus. Single cell suspensions were comprised of all major cell lineages found in the normal gastric glands. A method describing light scatter, size exclusion, doublet discrimination, viability staining, and fluorescently-conjugated antibodies and lectins was used to analyze individual epithelial cells and immune cells. This technique was capable of identifying parietal cells and revealed that gastric epithelial cells in the chronically inflamed mucosa significantly upregulated major histocompatibility complexes (MHC I and II but not CD80 or CD86, which are costimulatory molecules involved in T cell activation. These studies describe a method for isolating viable single cells and a detailed description of flow cytometric analysis of cells from healthy and diseased stomachs. These studies begin to identify effects of

Multidisciplinary System Reliability Analysis

Science.gov (United States)

Mahadevan, Sankaran; Han, Song; Chamis, Christos C. (Technical Monitor)

2001-01-01

The objective of this study is to develop a new methodology for estimating the reliability of engineering systems that encompass multiple disciplines. The methodology is formulated in the context of the NESSUS probabilistic structural analysis code, developed under the leadership of NASA Glenn Research Center. The NESSUS code has been successfully applied to the reliability estimation of a variety of structural engineering systems. This study examines whether the features of NESSUS could be used to investigate the reliability of systems in other disciplines such as heat transfer, fluid mechanics, electrical circuits etc., without considerable programming effort specific to each discipline. In this study, the mechanical equivalence between system behavior models in different disciplines are investigated to achieve this objective. A new methodology is presented for the analysis of heat transfer, fluid flow, and electrical circuit problems using the structural analysis routines within NESSUS, by utilizing the equivalence between the computational quantities in different disciplines. This technique is integrated with the fast probability integration and system reliability techniques within the NESSUS code, to successfully compute the system reliability of multidisciplinary systems. Traditional as well as progressive failure analysis methods for system reliability estimation are demonstrated, through a numerical example of a heat exchanger system involving failure modes in structural, heat transfer and fluid flow disciplines.

Resilience and reliability of compact vertical-flow treatment wetlands designed for tropical climates.

Science.gov (United States)

Lombard-Latune, R; Pelus, L; Fina, N; L'Etang, F; Le Guennec, B; Molle, P

2018-06-10

Most of the tropical areas have sanitation problems to contend with. The French system of vertical-flow treatment wetlands (FS-VFTW) fed with raw wastewater could be a good water and sludge management solution. The purpose-adapted tropical design can reduce area requirement to below 1 m 2 /population equivalents (p.e.). The Taupinière FS-VFTW on Martinique Island was built according to this design, with one stage but with a saturated layer at the bottom of the filter and a simplified trickling filter (TF) added for further treatment to meet the high performances targeted. Unsaturated/saturated vertical-flow filters (US/S FS-VFTW) have shown improved performances on total nitrogen, carbon and suspended solids removal in temperate climates, but the performances in tropical conditions remain unknown. Here, we report on real-world-operation in the French Overseas Territories (FOT), the reliability and performances of this VFCW tropical-design. The system experienced loading conditions ranging from 30% to 165% of nominal carbonaceous biological oxygen demand (BOD 5 ), as well as tropical rainstorms that brought over 7 times the nominal hydraulic load. Over a period of 3 years, 29 campaigns collected 24-h flow-proportional samples at each treatment stage (raw wastewater, FS-VFTW outlet, TF outlet). When applied loads were close to nominal values, the US/S FS-VFTW itself guarantees 85/90/60/50% removal and 125/25/40/50 mg/L at the outlet for chemical oxygen demand (COD)/total suspended solids (TSS)/total Kjeldahl nitrogen (TKN)/total nitrogen (TN), respectively. By comparison with US/S systems in mainland France, it appears that the warmer tropical-climate temperatures facilitate both nitrification and denitrification kinetics. Performances in overload conditions confirm that the US/S FS-VFTW remains robust and reliable although COD and TKN removal are impacted, especially after strong tropical rain events. By adding a simple compact trickling filter to a US/S FS

Prospective identification of erythroid elements in cultured peripheral blood.

Science.gov (United States)

Miller, J L; Njoroge, J M; Gubin, A N; Rodgers, G P

1999-04-01

We have developed a prospective approach to identify the generation of erythroid cells derived from cultured peripheral blood mononuclear cells (PBMC) by monitoring the expression of the cell surface protein CD48. Unpurified populations of PBMC obtained from the buffy coats of normal volunteers were grown in suspension culture in the absence or presence of erythropoietin. A profile of surface CD48 expression permitted a flow cytometric identification of erythropoietin responsive populations at various stages of their maturation. In the absence of erythropoietin (EPO) supplemented media, the CD48- cells represented <5% of the total population of PBMC remaining in culture. In cultures supplemented with 1 U/mL EPO, the mean percentage of CD48- cells increased to 34.7 + 14.9% (p < 0.01) after 14 days in culture. Coordinated CD34 and CD71 (transferrin receptor) expression, morphology, gamma-globin transcription, and colony formation in methylcellulose were observed during the 14-day culture period. Flow cytometric monitoring of bulk cultured PBMC provides a simple and reliable means for the prospective or real-time study of human erythropoiesis.

Whole blood flow cytometric analysis of Ureaplasma-stimulated monocytes from pregnant women.

Science.gov (United States)

Friedland, Yael D; Lee-Pullen, Tracey F; Nathan, Elizabeth; Watts, Rory; Keelan, Jeffrey A; Payne, Matthew S; Ireland, Demelza J

2015-06-01

We hypothesised that circulating monocytes of women with vaginal colonisation with Ureaplasma spp., genital microorganisms known to cause inflammation-driven preterm birth, would elicit a tolerised cytokine response to subsequent in vitro Ureaplasma parvum serovar 3 (UpSV3) stimulation. Using multi-parameter flow cytometry, we found no differences with regard to maternal colonisation status in the frequency of TNF-α-, IL-6-, IL-8- and IL-1β-expressing monocytes in response to subsequent UpSV3 stimulation (P > 0.10 for all cytokines). We conclude that vaginal Ureaplasma spp. colonisation does not specifically tolerise monocytes of pregnant women towards decreased responses to subsequent stimulation. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

Flow Cytometric Immunobead Assay for Detection of BCR-ABL1 Fusion Proteins in Chronic Myleoid Leukemia: Comparison with FISH and PCR Techniques

Science.gov (United States)

Recchia, Anna Grazia; Caruso, Nadia; Bossio, Sabrina; Pellicanò, Mariavaleria; De Stefano, Laura; Franzese, Stefania; Palummo, Angela; Abbadessa, Vincenzo; Lucia, Eugenio; Gentile, Massimo; Vigna, Ernesto; Caracciolo, Clementina; Agostino, Antolino; Galimberti, Sara; Levato, Luciano; Stagno, Fabio; Molica, Stefano; Martino, Bruno; Vigneri, Paolo; Di Raimondo, Francesco; Morabito, Fortunato

2015-01-01

Chronic Myeloid Leukemia (CML) is characterized by a balanced translocation juxtaposing the Abelson (ABL) and breakpoint cluster region (BCR) genes. The resulting BCR-ABL1 oncogene leads to increased proliferation and survival of leukemic cells. Successful treatment of CML has been accompanied by steady improvements in our capacity to accurately and sensitively monitor therapy response. Currently, measurement of BCR-ABL1 mRNA transcript levels by real-time quantitative PCR (RQ-PCR) defines critical response endpoints. An antibody-based technique for BCR-ABL1 protein recognition could be an attractive alternative to RQ-PCR. To date, there have been no studies evaluating whether flow-cytometry based assays could be of clinical utility in evaluating residual disease in CML patients. Here we describe a flow-cytometry assay that detects the presence of BCR-ABL1 fusion proteins in CML lysates to determine the applicability, reliability, and specificity of this method for both diagnosis and monitoring of CML patients for initial response to therapy. We show that: i) CML can be properly diagnosed at onset, (ii) follow-up assessments show detectable fusion protein (i.e. relative mean fluorescent intensity, rMFI%>1) when BCR-ABL1IS transcripts are between 1–10%, and (iii) rMFI% levels predict CCyR as defined by FISH analysis. Overall, the FCBA assay is a rapid technique, fully translatable to the routine management of CML patients. PMID:26111048

Flow Cytometric Immunobead Assay for Detection of BCR-ABL1 Fusion Proteins in Chronic Myleoid Leukemia: Comparison with FISH and PCR Techniques.

Directory of Open Access Journals (Sweden)

Anna Grazia Recchia

Full Text Available Chronic Myeloid Leukemia (CML is characterized by a balanced translocation juxtaposing the Abelson (ABL and breakpoint cluster region (BCR genes. The resulting BCR-ABL1 oncogene leads to increased proliferation and survival of leukemic cells. Successful treatment of CML has been accompanied by steady improvements in our capacity to accurately and sensitively monitor therapy response. Currently, measurement of BCR-ABL1 mRNA transcript levels by real-time quantitative PCR (RQ-PCR defines critical response endpoints. An antibody-based technique for BCR-ABL1 protein recognition could be an attractive alternative to RQ-PCR. To date, there have been no studies evaluating whether flow-cytometry based assays could be of clinical utility in evaluating residual disease in CML patients. Here we describe a flow-cytometry assay that detects the presence of BCR-ABL1 fusion proteins in CML lysates to determine the applicability, reliability, and specificity of this method for both diagnosis and monitoring of CML patients for initial response to therapy. We show that: i CML can be properly diagnosed at onset, (ii follow-up assessments show detectable fusion protein (i.e. relative mean fluorescent intensity, rMFI%>1 when BCR-ABL1IS transcripts are between 1-10%, and (iii rMFI% levels predict CCyR as defined by FISH analysis. Overall, the FCBA assay is a rapid technique, fully translatable to the routine management of CML patients.

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.

Flow cytometry with gold nanoparticles and their clusters as scattering contrast agents: FDTD simulation of light-cell interaction.

Science.gov (United States)

Tanev, Stoyan; Sun, Wenbo; Pond, James; Tuchin, Valery V; Zharov, Vladimir P

2009-09-01

The formulation of the finite-difference time-domain (FDTD) approach is presented in the framework of its potential applications to in-vivo flow cytometry based on light scattering. The consideration is focused on comparison of light scattering by a single biological cell alone in controlled refractive-index matching conditions and by cells labeled by gold nanoparticles. The optical schematics including phase contrast (OPCM) microscopy as a prospective modality for in-vivo flow cytometry is also analyzed. The validation of the FDTD approach for the simulation of flow cytometry may open up a new avenue in the development of advanced cytometric techniques based on scattering effects from nanoscale targets. 2009 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

Intra-Prostate Cancer Vaccine Inducer

Science.gov (United States)

2006-07-01

4 that RNAi technology is possibly a more effective and reliable method to silence expression of a given gene. Therefore, we constructed an Ii...experiments, bone marrow cells were extracted from the femurs of BALB/c mice. Cells were plated at 4x106 cells in 100 mm dishes, in RPMI 1640-medium...permeabilized with saponin in preparation for staining with Ii monoclonal antibodies (data not shown). C1 E1 C2 E2 Figure 3 (C1) represents flow cytometric

Multi-Disciplinary System Reliability Analysis

Science.gov (United States)

Mahadevan, Sankaran; Han, Song

1997-01-01

The objective of this study is to develop a new methodology for estimating the reliability of engineering systems that encompass multiple disciplines. The methodology is formulated in the context of the NESSUS probabilistic structural analysis code developed under the leadership of NASA Lewis Research Center. The NESSUS code has been successfully applied to the reliability estimation of a variety of structural engineering systems. This study examines whether the features of NESSUS could be used to investigate the reliability of systems in other disciplines such as heat transfer, fluid mechanics, electrical circuits etc., without considerable programming effort specific to each discipline. In this study, the mechanical equivalence between system behavior models in different disciplines are investigated to achieve this objective. A new methodology is presented for the analysis of heat transfer, fluid flow, and electrical circuit problems using the structural analysis routines within NESSUS, by utilizing the equivalence between the computational quantities in different disciplines. This technique is integrated with the fast probability integration and system reliability techniques within the NESSUS code, to successfully compute the system reliability of multi-disciplinary systems. Traditional as well as progressive failure analysis methods for system reliability estimation are demonstrated, through a numerical example of a heat exchanger system involving failure modes in structural, heat transfer and fluid flow disciplines.

Flow cytometric monitoring of influenza A virus infection in MDCK cells during vaccine production

Directory of Open Access Journals (Sweden)

Reichl Udo

2008-04-01

Full Text Available Abstract Background In cell culture-based influenza vaccine production the monitoring of virus titres and cell physiology during infection is of great importance for process characterisation and optimisation. While conventional virus quantification methods give only virus titres in the culture broth, data obtained by fluorescence labelling of intracellular virus proteins provide additional information on infection dynamics. Flow cytometry represents a valuable tool to investigate the influences of cultivation conditions and process variations on virus replication and virus yields. Results In this study, fluorescein-labelled monoclonal antibodies against influenza A virus matrix protein 1 and nucleoprotein were used for monitoring the infection status of adherent Madin-Darby canine kidney cells from bioreactor samples. Monoclonal antibody binding was shown for influenza A virus strains of different subtypes (H1N1, H1N2, H3N8 and host specificity (human, equine, swine. At high multiplicity of infection in a bioreactor, the onset of viral protein accumulation in adherent cells on microcarriers was detected at about 2 to 4 h post infection by flow cytometry. In contrast, a significant increase in titre by hemagglutination assay was detected at the earliest 4 to 6 h post infection. Conclusion It is shown that flow cytometry is a sensitive and robust method for the monitoring of viral infection in fixed cells from bioreactor samples. Therefore, it is a valuable addition to other detection methods of influenza virus infection such as immunotitration and RNA hybridisation. Thousands of individual cells are measured per sample. Thus, the presented method is believed to be quite independent of the concentration of infected cells (multiplicity of infection and total cell concentration in bioreactors. This allows to perform detailed studies on factors relevant for optimization of virus yields in cell cultures. The method could also be used for process

Analysis of the Budding Yeast Cell Cycle by Flow Cytometry.

Science.gov (United States)

Rosebrock, Adam P

2017-01-03

DNA synthesis is one of the landmark events in the cell cycle: G 1 cells have one copy of the genome, S phase cells are actively engaged in DNA synthesis, and G 2 cells have twice as much nuclear DNA as G 1 cells. Cellular DNA content can be measured by staining with a fluorescent dye followed by a flow-cytometric readout. This method provides a quantitative measurement of cell cycle position on a cell-by-cell basis at high speed. Using flow cytometry, tens of thousands of single-cell measurements can be generated in a few seconds. This protocol details staining of cells of the budding yeast Saccharomyces cerevisiae for flow cytometry using Sytox Green dye in a method that can be scaled widely-from one sample to many thousands and operating on inputs ranging from 1 million to more than 100 million cells. Flow cytometry is preferred over light microscopy or Coulter analyses for the analysis of the cell cycle as DNA content and cell cycle position are being directly measured. © 2017 Cold Spring Harbor Laboratory Press.

Isolation and characterization of mouse innate lymphoid cells.

Science.gov (United States)

Halim, Timotheus Y F; Takei, Fumio

2014-08-01

Innate lymphoid cells (ILCs) are rare populations of cytokine-producing lymphocytes and are divided into three groups, namely ILC1, ILC2, and ILC3, based on the cytokines that they produce. They comprise less than 1% of lymphocytes in mucosal tissues and express no unique cell surface markers. Therefore, they can only be identified by combinations of multiple cell surface markers and further characterized by cytokine production in vitro. Thus, multicolor flow cytometry is the only reliable method to purify and characterize ILCs. Here we describe the methods for cell preparation, flow cytometric analysis, and purification of murine ILC2 and ILC3. Copyright © 2014 John Wiley & Sons, Inc.

Quality assessment program for EuroFlow protocols: summary results of four-year (2010-2013) quality assurance rounds.

Science.gov (United States)

Kalina, Tomas; Flores-Montero, Juan; Lecrevisse, Quentin; Pedreira, Carlos E; van der Velden, Vincent H J; Novakova, Michaela; Mejstrikova, Ester; Hrusak, Ondrej; Böttcher, Sebastian; Karsch, Dennis; Sędek, Łukasz; Trinquand, Amelie; Boeckx, Nancy; Caetano, Joana; Asnafi, Vahid; Lucio, Paulo; Lima, Margarida; Helena Santos, Ana; Bonaccorso, Paola; van der Sluijs-Gelling, Alita J; Langerak, Anton W; Martin-Ayuso, Marta; Szczepański, Tomasz; van Dongen, Jacques J M; Orfao, Alberto

2015-02-01

Flow cytometric immunophenotyping has become essential for accurate diagnosis, classification, and disease monitoring in hemato-oncology. The EuroFlow Consortium has established a fully standardized "all-in-one" pipeline consisting of standardized instrument settings, reagent panels, and sample preparation protocols and software for data analysis and disease classification. For its reproducible implementation, parallel development of a quality assurance (QA) program was required. Here, we report on the results of four consecutive annual rounds of the novel external QA EuroFlow program. The novel QA scheme aimed at monitoring the whole flow cytometric analysis process (cytometer setting, sample preparation, acquisition and analysis) by reading the median fluorescence intensities (MedFI) of defined lymphocytes' subsets. Each QA participant applied the predefined reagents' panel on blood cells of local healthy donors. A uniform gating strategy was applied to define lymphocyte subsets and to read MedFI values per marker. The MedFI values were compared with reference data and deviations from reference values were quantified using performance score metrics. In four annual QA rounds, we analyzed 123 blood samples from local healthy donors on 14 different instruments in 11 laboratories from nine European countries. The immunophenotype of defined cellular subsets appeared sufficiently standardized to permit unified (software) data analysis. The coefficient of variation of MedFI for 7 of 11 markers performed repeatedly below 30%, average MedFI in each QA round ranged from 86 to 125% from overall median. Calculation of performance scores was instrumental to pinpoint standardization failures and their causes. Overall, the new EuroFlow QA system for the first time allowed to quantify the technical variation that is introduced in the measurement of fluorescence intensities in a multicentric setting over an extended period of time. EuroFlow QA is a proficiency test specific for

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.

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.

Flow cytometric analysis of microbial contamination in food industry technological lines – initial study

OpenAIRE

Katarzyna Czaczyk; Wojciech Juzwa

2012-01-01

Background. Flow cytometry constitutes an alternative for traditional methods of microorganisms identifi cation 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 signifi cant reduction of time required for analysis allowing monitoring of production process...

Transmission cost allocation based on power flow tracing considering reliability benefit

International Nuclear Information System (INIS)

Leepreechanon, N.; Singharerg, S.; Padungwech, W.; Nakawiro, W.; Eua-Arporn, B.; David, A.K.

2007-01-01

Power transmission networks must be able to accommodate the continuously growing demand for reliable and economical electricity. This paper presented a method to allocate transmission use and reliability cost to both generators and end-consumers. Although transmission cost allocation methods change depending on the local context of the electric power industry, there is a common principle that transmission line capacity should be properly allocated to accommodate actual power delivery with an adequate reliability margin. The method proposed in this paper allocates transmission embedded cost to both generators and loads in an equitable manner, incorporating probability indices to allocate transmission reliability margin among users in both supply and demand sides. The application of the proposed method was illustrated using Bialek's tracing method on a multiple-circuit, six-bus transmission system. Probabilistic indices known as the transmission internal reliability margin (TIRM) and transmission external reliability margin (TERM) decomposed from the transmission reliability margin (TRM) were introduced, making true cost of using overall transmission facilities. 6 refs., 11 tabs., 5 figs

Flow cytometry detection of planktonic cells with polycyclic aromatic hydrocarbons sorbed to cell surfaces

KAUST Repository

Cerezo, Maria I.

2017-02-17

Polycyclic aromatic hydrocarbons are very important components of oil pollution. These pollutants tend to sorb to cell surfaces, exerting toxic effects on organisms. Our study developed a flow cytometric method for the detection of PAHs sorbed to phytoplankton by exploiting their spectral characteristics. We discriminated between cells with PAHs from cells free of PAHs. Clear discrimination was observed with flow cytometer provided with 375 or 405nm lasers in addition to the standard 488nm laser necessary to identify phytoplankton. Using this method, we measured the relationship between the percentages of phytoplankton organisms with PAHs, with the decrease in the growth rate. Moreover, the development of this method could be extended to facilitate the study of PAHs impact on cell cultures from a large variety of organisms.

Reliability and effectiveness of early warning systems for natural hazards: Concept and application to debris flow warning

International Nuclear Information System (INIS)

Sättele, Martina; Bründl, Michael; Straub, Daniel

2015-01-01

Early Warning Systems (EWS) are increasingly applied to mitigate the risks posed by natural hazards. To compare the effect of EWS with alternative risk reduction measures and to optimize their design and operation, their reliability and effectiveness must be quantified. In the present contribution, a framework approach to the evaluation of threshold-based EWS for natural hazards is presented. The system reliability is classically represented by the Probability of Detection (POD) and Probability of False Alarms (PFA). We demonstrate how the EWS effectiveness, which is a measure of risk reduction, can be formulated as a function of POD and PFA. To model the EWS and compute the reliability, we develop a framework based on Bayesian Networks, which is further extended to a decision graph, facilitating the optimization of the warning system. In a case study, the framework is applied to the assessment of an existing debris flow EWS. The application demonstrates the potential of the framework for identifying the important factors influencing the effectiveness of the EWS and determining optimal warning strategies and system configurations. - Highlights: • Warning systems are increasingly applied measures to reduce natural hazard risks. • Bayesian Networks (BN) are powerful tools to quantify warning system's reliability. • The effectiveness is defined to assess the optimality of warning systems. • By extending BNs to decision graphs, the optimal warning strategy is identified. • Sensors positioning significantly influence the effectiveness of warning systems

New Approaches to Reliability Assessment

DEFF Research Database (Denmark)

Ma, Ke; Wang, Huai; Blaabjerg, Frede

2016-01-01

of energy. New approaches for reliability assessment are being taken in the design phase of power electronics systems based on the physics-of-failure in components. In this approach, many new methods, such as multidisciplinary simulation tools, strength testing of components, translation of mission profiles......, and statistical analysis, are involved to enable better prediction and design of reliability for products. This article gives an overview of the new design flow in the reliability engineering of power electronics from the system-level point of view and discusses some of the emerging needs for the technology...

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

International Nuclear Information System (INIS)

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

2003-01-01

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

Flow cytometric analysis of regulatory T cells during hyposensitization of acquired allergic contact dermatitis.

Science.gov (United States)

Fraser, Kathleen; Abbas, Mariam; Hull, Peter R

2014-01-01

We previously demonstrated that repeated intradermal steroid injections administered at weekly intervals into positive patch-test sites induce hyposensitization and desensitization. To examine changes in CD4CD25CD127lo/ regulatory T cells during the attenuation of the patch-test response. Ten patients with known allergic contact dermatitis were patch tested weekly for 10 weeks. The patch-test site was injected intradermally with 2 mg triamcinolone. At weeks 1 and 7, a biopsy was performed on the patch-test site in 6 patients, and flow cytometry was performed assessing CD4CD25CD127lo/ regulatory T cells. Secondary outcomes were clinical score, reaction size, erythema, and temperature. Statistical analysis included regression, correlation, and repeated-measures analysis of variance. The percentage of CD4CD25CD127lo/ regulatory T cells, measured by flow cytometry, increased from week 1 to week 7 by an average of 19.2%. The average grade of patch-test reaction decreased from +++ (vesicular reaction) to ++ (palpable erythema). The mean drop in temperature following treatment was 0.28°C per week. The mean area decreased 8.6 mm/wk over 10 weeks. Intradermal steroid injections of weekly patch-test reactions resulted in hyposensitization of the allergic contact dermatitis reaction. CD4CD25CD127lo/ regulatory T cells showed a tendency to increase; however, further studies are needed to determine if this is significant.

Synthesis of new, UV-photoactive dansyl derivatives for flow cytometric studies on bile acid uptake.

Science.gov (United States)

Rohacova, Jana; Marin, M Luisa; Martínez-Romero, Alicia; O'Connor, José-Enrique; Gomez-Lechon, M Jose; Donato, M Teresa; Castell, Jose V; Miranda, Miguel A

2009-12-07

Four new fluorescent derivatives of cholic acid have been synthesized; they incorporate a dansyl moiety at 3alpha-, 3beta-, 7alpha- or 7beta- positions. These cholic acid analogs are UV photoactive and also exhibit green fluorescence. In addition, they have been demonstrated to be suitable for studying the kinetics of bile acid transport by flow cytometry.

Construction of BAC Libraries from Flow-Sorted Chromosomes.

Science.gov (United States)

Šafář, Jan; Šimková, Hana; Doležel, Jaroslav

2016-01-01

Cloned DNA libraries in bacterial artificial chromosome (BAC) are the most widely used form of large-insert DNA libraries. BAC libraries are typically represented by ordered clones derived from genomic DNA of a particular organism. In the case of large eukaryotic genomes, whole-genome libraries consist of a hundred thousand to a million clones, which make their handling and screening a daunting task. The labor and cost of working with whole-genome libraries can be greatly reduced by constructing a library derived from a smaller part of the genome. Here we describe construction of BAC libraries from mitotic chromosomes purified by flow cytometric sorting. Chromosome-specific BAC libraries facilitate positional gene cloning, physical mapping, and sequencing in complex plant genomes.

Flow cytometric measurement of DNA level and steroid hormone receptor assay in breast cancer

International Nuclear Information System (INIS)

Zubrikhina, G.N.; Kuz'mina, Eh.V.; Bassalyk, L.S.; Murav'eva, N.I.

1989-01-01

DNA level measured by flow cytometry and estrogen and progesteron receptors assayed in tissue samples obtained from 85 malignant and 16 benign lesions of the breast. All the benign tumors revealed 2c DNA content and most of them were receptor-negative, while 74.1% of breast carcinomas displayed aneuploidy. Three patients (3.5%) had two lines of aneuploid cells. Many aneuploid tumors were receptor-negative. Preoperative radiation treatmet (14-20 Gy) did not significantly influence the level of steroid hormone receptors in tumors. Estrogen receptor level was higher in menopausal patients than in premenopausal ones

Comparative study on 4 quantitative detection methods of apoptosis induced by radiation

International Nuclear Information System (INIS)

Yang Yepeng; Chen Guanying; Zhou Mei; Shen Qinjian; Shen Lei; Zhu Yingbao

2004-01-01

Objective: To reveal the capability of 4 apoptosis-detecting methods to discriminate between apoptosis and necrosis and show their respective advantages and shortcomings through comparison of detected results and analysis of detection mechanism. Methods: Four methods, PI staining-flow cytometric detection (P-F method), TUNEL labeling-flow cytometric detection (T-F method), annexing V-FITC/PI vital staining-flow cytometric detection (A-F method) and Hoechst/PI vital staining-fluorescence microscopic observation (H-O method), were used to determine apoptosis and necrosis in human breast cancer MCF-7 cell line induced by γ-rays. Hydroxycamptothecine and sodium azide were used to induce positive controls of apoptosis and necrosis respectively. Results: All 4 methods showed good time-dependent and dose dependent respondence to apoptosis induced by γ-rays and hydroxycamptothecine. Apoptotic cell ratios and curve slopes obtained from P-F method were minimum and, on the contrary, those from T-F method were maximum among these 4 methods. With A-F method and H-O method, two sets of data, apoptosis and necrosis, could be gained respectively and the data gained from these two methods were close to equal. A-F method and H-O method could distinguish necrosis induced by sodium azide from apoptosis while P-F method and T-F method presented false increase of apoptosis. Conclusions: P-F method and T-F method can not discriminate between apoptosis and necrosis. P-F method is less sensitive but more simple, convenient and economical than T-F method. A-F method and H-O method can distinguish necrosis from apoptosis. A-F method is more costly but more quick and reliable than H-O method. H-O method is economical, practical and morphological changes of cells and nucleus can be observed simultaneously with it. (authors)

Antibody-modified iron oxide nanoparticles for efficient magnetic isolation and flow cytometric determination of L. pneumophila

International Nuclear Information System (INIS)

Bloemen, Maarten; Verbiest, Thierry; Denis, Carla; Meester, Luc De; Peeters, Miet; Gils, Ann; Geukens, Nick

2015-01-01

We report on the design of superparamagnetic nanoparticles capable of selectively isolating targeted bacteria (Legionella pneumophila, serogroup 1) from aqueous solutions. The surface of magnetite nanoparticles (NP) was functionalized with a heterobifunctional poly(ethylene glycol) ligand containing reactive groups for covalent coupling of polyclonal antibodies against L. pneumophila. These bioconjugates were used to label and magnetically isolate L. pneumophila. Flow cytometry revealed high separation and efficiency in this regard. The strain specificity and efficiency of the magnetic NP was tested with recombinant strains of E. coli (expressing the red fluorescent protein) and L. pneumophila (expressing the green fluorescent protein). The detection limit of the method (by flow cytometry) is 10 4 cells∙mL -1 . The results indicate that the new multifunctional NPs are capable of selectively attracting pathogens from a complex mixture and with high efficiency. This, conceivably, paves the way to pre-concentration protocols for numerous other pathogens. (author)

14CO2 labeling. A reliable technique for rapid measurement of total root exudation capacity and vascular sap flow in crop plants

International Nuclear Information System (INIS)

Bhupinder Singh; Sumedha Ahuja; Renu Pandey; Singhal, R.K.

2014-01-01

Root release of organic compounds and rate of the vascular sap flow are important for understanding the nutrient and the source-sink dynamics in plants, however, their determination is procedurally cumbersome and time consuming. We report here a simple method involving 14 C labeling for rapid and reliable measurement of root exudates and vascular sap flow rate in a variable groundnut population developed through seed gamma irradiation using a cobalt source ( 60 Co). An experimental hypothesis that a higher 14 C level in the vascular sap would indicate a higher root release of carbon by the roots into the rhizosphere was verified. (author)

Challenges Regarding IP Core Functional Reliability

Science.gov (United States)

Berg, Melanie D.; LaBel, Kenneth A.

2017-01-01

For many years, intellectual property (IP) cores have been incorporated into field programmable gate array (FPGA) and application specific integrated circuit (ASIC) design flows. However, the usage of large complex IP cores were limited within products that required a high level of reliability. This is no longer the case. IP core insertion has become mainstream including their use in highly reliable products. Due to limited visibility and control, challenges exist when using IP cores and subsequently compromise product reliability. We discuss challenges and suggest potential solutions to critical application IP insertion.

Sex-sorting sperm using flow cytometry/cell sorting.

Science.gov (United States)

Garner, Duane L; Evans, K Michael; Seidel, George E

2013-01-01

The sex of mammalian offspring can be predetermined by flow sorting relatively pure living populations of X- and Y-chromosome-bearing sperm. This method is based on precise staining of the DNA of sperm with the nucleic acid-specific fluorophore, Hoechst 33342, to differentiate between the subpopulations of X- and Y-sperm. The fluorescently stained sperm are then sex-sorted using a specialized high speed sorter, MoFlo(®) SX XDP, and collected into biologically supportive media prior to reconcentration and cryopreservation in numbers adequate for use with artificial insemination for some species or for in vitro fertilization. Sperm sorting can provide subpopulations of X- or Y-bearing bovine sperm at rates in the 8,000 sperm/s range while maintaining; a purity of 90% such that it has been applied to cattle on a commercial basis. The sex of offspring has been predetermined in a wide variety of mammalian species including cattle, swine, horses, sheep, goats, dogs, cats, deer, elk, dolphins, water buffalo as well as in humans using flow cytometric sorting of X- and Y-sperm.

Flow cytometric evaluation of antibiotic effects on viability and mitochondrial function of refrigerated spermatozoa of Nile tilapia

Science.gov (United States)

Segovia, M.; Jenkins, J.A.; Paniagua-Chavez, C.; Tiersch, T.R.

2000-01-01

Improved techniques for storage and evaluation of fish sperm would enhance breeding programs around the world. The goal of this study was to test the effect of antibiotics on refrigerated sperm from Nile tilapia (Oreochromis niloticus) by use of flow cytometry with 2 dual-staining protocols for objective assessment of sperm quality. Concentrations of 1 x 109 sperm/mL were suspended in Ringer's buffer at 318 mOsmol/kg (pH 8.0). The fluorescent stains Sybr 14 (10 ??M), propidium iodide (2.4 mM), and rhodamine 123 (0.13 ??M) were used to assess cell viability and mitochondrial function. Three concentrations of ampicillin, gentamicin, and an antibiotic/antimycotic solution were added to fresh spermatozoa. Motility estimates and flow cytometry measurements were made daily during 7 d of refrigerated storage (4 ??C). The highest concentrations of gentamicin and antibiotic/antimycotic and all 3 concentrations of ampicillin significantly reduced sperm viability. The highest of each of the 3 antibiotic concentrations significantly reduced mitochondrial function. This study demonstrates that objective sperm quality assessments can be made using flow cytometry and that addition of antibiotics at appropriate concentrations can lengthen refrigerated storage time for tilapia spermatozoa. With minor modifications, these protocols can be adapted for use with sperm from other species and with other tissue types.

75 FR 15371 - Time Error Correction Reliability Standard

Science.gov (United States)

2010-03-29

... Electric Reliability Council of Texas (ERCOT) manages the flow of electric power to 22 million Texas customers. As the independent system operator for the region, ERCOT schedules power on an electric grid that... Coordinating Council (WECC) is responsible for coordinating and promoting bulk electric system reliability in...

Dynamic reliability networks with self-healing units

International Nuclear Information System (INIS)

Jenab, K.; Seyed Hosseini, S.M.; Dhillon, B.S.

2008-01-01

This paper presents an analytical approach for dynamic reliability networks used for the failure limit strategy in maintenance optimization. The proposed approach utilizes the moment generating function (MGF) and the flow-graph concept to depict the functional and reliability diagrams of the system comprised of series, parallel or mix configuration of self-healing units. The self-healing unit is featured by the embedded failure detection and recovery mechanisms presented by self-loop in flow-graph networks. The newly developed analytical approach provides the probability of the system failure and time-to-failure data i.e., mean and standard deviation time-to-failure used for maintenance optimization

A complementary role of multiparameter flow cytometry and high-throughput sequencing for minimal residual disease detection in chronic lymphocytic leukemia: an European Research Initiative on CLL study.

LENUS (Irish Health Repository)

Rawstron, A C

2016-04-01

In chronic lymphocytic leukemia (CLL) the level of minimal residual disease (MRD) after therapy is an independent predictor of outcome. Given the increasing number of new agents being explored for CLL therapy, using MRD as a surrogate could greatly reduce the time necessary to assess their efficacy. In this European Research Initiative on CLL (ERIC) project we have identified and validated a flow-cytometric approach to reliably quantitate CLL cells to the level of 0.0010% (10(-5)). The assay comprises a core panel of six markers (i.e. CD19, CD20, CD5, CD43, CD79b and CD81) with a component specification independent of instrument and reagents, which can be locally re-validated using normal peripheral blood. This method is directly comparable to previous ERIC-designed assays and also provides a backbone for investigation of new markers. A parallel analysis of high-throughput sequencing using the ClonoSEQ assay showed good concordance with flow cytometry results at the 0.010% (10(-4)) level, the MRD threshold defined in the 2008 International Workshop on CLL guidelines, but it also provides good linearity to a detection limit of 1 in a million (10(-6)). The combination of both technologies would permit a highly sensitive approach to MRD detection while providing a reproducible and broadly accessible method to quantify residual disease and optimize treatment in CLL.

76 FR 23470 - Version One Regional Reliability Standard for Transmission Operations

Science.gov (United States)

2011-04-27

...; Order No. 752] Version One Regional Reliability Standard for Transmission Operations AGENCY: Federal... Power Act, the Federal Energy Regulatory Commission approves regional Reliability Standard TOP-007-WECC... purpose of this regional Reliability Standard is to ensure that actual flows and associated scheduled...

Biology and flow cytometry of proangiogenic hematopoietic progenitors cells.

Science.gov (United States)

Rose, Jonathan A; Erzurum, Serpil; Asosingh, Kewal

2015-01-01

During development, hematopoiesis and neovascularization are closely linked to each other via a common bipotent stem cell called the hemangioblast that gives rise to both hematopoietic cells and endothelial cells. In postnatal life, this functional connection between the vasculature and hematopoiesis is maintained by a subset of hematopoietic progenitor cells endowed with the capacity to differentiate into potent proangiogenic cells. These proangiogenic hematopoietic progenitors comprise a specific subset of bone marrow (BM)-derived cells that homes to sites of neovascularization and possess potent paracrine angiogenic activity. There is emerging evidence that this subpopulation of hematopoietic progenitors plays a critical role in vascular health and disease. Their angiogenic activity is distinct from putative "endothelial progenitor cells" that become structural cells of the endothelium by differentiation into endothelial cells. Proangiogenic hematopoietic progenitor cell research requires multidisciplinary expertise in flow cytometry, hematology, and vascular biology. This review provides a comprehensive overview of proangiogenic hematopoietic progenitor cell biology and flow cytometric methods to detect these cells in the peripheral blood circulation and BM. © 2014 International Society for Advancement of Cytometry.

Determination of Complement-Mediated Killing of Bacteria by Viability Staining and Bioluminescence

OpenAIRE

Virta, Marko; Lineri, Sanna; Kankaanpää, Pasi; Karp, Matti; Peltonen, Karita; Nuutila, Jari; Lilius, Esa-Matti

1998-01-01

Complement-mediated killing of bacteria was monitored by flow cytometric, luminometric, and conventional plate counting methods. A flow cytometric determination of bacterial viability was carried out by using dual staining with a LIVE/DEAD BacLight bacterial viability kit. In addition to the viable cell population, several other populations emerged in the fluorescence histogram, and there was a dramatic decrease in the total cell count in the light-scattering histogram in the course of the co...

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.

Flow-cytometric determination of high-density-lipoprotein binding sites on human leukocytes

International Nuclear Information System (INIS)

Schmitz, G.; Wulf, G.; Bruening, T.A.; Assmann, G.

1987-01-01

In this method, leukocytes were isolated from 6 mL of EDTA-blood by density-gradient centrifugation and subsequently incubated with rhodamine isothiocyanate (RITC)-conjugated high-density lipoproteins (HDL). The receptor-bound conjugate particles were determined by fluorescent flow cytometry and compared with 125 I-labeled HDL binding data for the same cells. Human granulocytes express the highest number of HDL binding sites (9.4 x 10(4)/cell), followed by monocytes (7.3 x 10(4)/cell) and lymphocytes (4.0 x 10(4)/cell). Compared with conventional analysis of binding of 125 I-labeled HDL in tissue-culture dishes, the present determination revealed significantly lower values for nonspecific binding. In competition studies, the conjugate competes for the same binding sites as 125 I-labeled HDL. With the use of tetranitromethane-treated HDL3, which fails to compete for the HDL receptor sites while nonspecific binding is not affected, we could clearly distinguish between 37 degrees C surface binding and specific 37 degrees C uptake of RITC-HDL3, confirming that the HDL receptor leads bound HDL particles into an intracellular pathway rather than acting as a docking type of receptor. Patients with familial dysbetalipoproteinemia showed a significantly higher number of HDL binding sites in the granulocyte population but normal in lymphocytes and monocytes, indicating increased uptake of cholesterol-containing lipoproteins. In patients with familial hypercholesterolemia, HDL binding was increased in all three cell types, indicating increased cholesterol uptake and increased cholesterol synthesis. The present method allows rapid determination of HDL binding sites in leukocytes from patients with various forms of hyper- and dyslipoproteinemias

Reliability of laser Doppler, near-infrared spectroscopy and Doppler ultrasound for peripheral blood flow measurements during and after exercise in the heat.

Science.gov (United States)

Choo, Hui C; Nosaka, Kazunori; Peiffer, Jeremiah J; Ihsan, Mohammed; Yeo, Chow C; Abbiss, Chris R

2017-09-01

This study examined the test-retest reliability of near-infrared spectroscopy (NIRS), laser Doppler flowmetry (LDF) and Doppler ultrasound to assess exercise-induced haemodynamics. Nine men completed two identical trials consisting of 25-min submaximal cycling at first ventilatory threshold followed by repeated 30-s bouts of high-intensity (90% of peak power) cycling in 32.8 ± 0.4°C and 32 ± 5% relative humidity (RH). NIRS (tissue oxygenation index [TOI] and total haemoglobin [tHb]) and LDF (perfusion units [PU]) signals were monitored continuously during exercise, and leg blood flow was assessed by Doppler ultrasound at baseline and after exercise. Cutaneous vascular conductance (CVC; PU/mean arterial pressure (MAP)) was expressed as the percentage change from baseline (%CVC BL ). Coefficients of variation (CVs) as indicators of absolute reliability were 18.7-28.4%, 20.2-33.1%, 42.5-59.8%, 7.8-12.4% and 22.2-30.3% for PU, CVC, %CVC BL , TOI and tHb, respectively. CVs for these variables improved as exercise continued beyond 10 min. CVs for baseline and post-exercise leg blood flow were 17.8% and 10.5%, respectively. CVs for PU, tHb (r 2  = 0.062) and TOI (r 2  = 0.002) were not correlated (P > 0.05). Most variables demonstrated CVs lower than the expected changes (35%) induced by training or heat stress; however, minimum of 10 min exercise is recommended for more reliable measurements.

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)

Induction of chromosome damage by ultraviolet light and caffeine: correlation of cytogenetic evaluation and flow karyotype

International Nuclear Information System (INIS)

Cremer, C.; Cremer, T.; Gray, J.W.

1982-01-01

Asynchronously growing cells of a M3-1 Chinese hamster line were ultraviolet (UV) irradiated (lambda . 254 nm) with UV fluences up to 7.5 J/m(2). After irradiation cells were incubated with or without 2 mM caffeine for 20 hr, then mitotic cells were selected by mechanical shaking. Their chromosomes were isolated, stained with Hoechst 33258 and chromomycin A3, and measured flow cytometrically. While the fluorescence distributions of chromosomes (flow karyo-types) from cells treated with UV alone or with caffeine alone were very similar to those of untreated controls, the flow karyo-types of UV + caffeine-treated cells showed a debris continuum that increased with increasing UV fluence suggesting an increased number of chromosome fragments. Visual evaluation of metaphase plates revealed that the percentage of cells with chromosome damage also increased steadily with increasing UV fluence. A high degree of correlation was observed between the relative magnitude of the debris level from flow karyotypes and the percentage of cells with chromosome damage and with generalized chromosomes shattering, respectively, as determined from metaphase spreads

High reliability flow system - an assessment of pump reliability and optimisation of the number of pumps

International Nuclear Information System (INIS)

Butterfield, J.M.

1981-01-01

A system is considered where a number of pumps operate in parallel. Normally, all pumps operate, driven by main motors fed from the grid. Each pump has a pony motor fed from an individual battery supply. Each pony motor is normally running, but not engaged to the pump shaft. On demand, e.g. failure of grid supplies, each pony motor is designed to clutch-in automatically when the pump speed falls to a specified value. The probability of all the pony motors failing to clutch-in on demand must be demonstrated with 95% confidence to be less than 10 -8 per demand. This assessment considers how the required reliability of pony motor drives might be demonstrated in practice and the implications on choice of the number of pumps at the design stage. The assessment recognises that not only must the system prove to be extremely reliable, but that demonstration that reliability is adequate must be done during plant commissioning, with practical limits on the amount of testing performed. It is concluded that a minimum of eight pony motors should be provided, eight pumps each with one pony motor (preferred) or five pumps each with two independent pony motors. A minimum of two diverse pony motor systems should be provided. (author)

Automated Blood Sample Preparation Unit (ABSPU) for Portable Microfluidic Flow Cytometry.

Science.gov (United States)

Chaturvedi, Akhil; Gorthi, Sai Siva

2017-02-01

Portable microfluidic diagnostic devices, including flow cytometers, are being developed for point-of-care settings, especially in conjunction with inexpensive imaging devices such as mobile phone cameras. However, two pervasive drawbacks of these have been the lack of automated sample preparation processes and cells settling out of sample suspensions, leading to inaccurate results. We report an automated blood sample preparation unit (ABSPU) to prevent blood samples from settling in a reservoir during loading of samples in flow cytometers. This apparatus automates the preanalytical steps of dilution and staining of blood cells prior to microfluidic loading. It employs an assembly with a miniature vibration motor to drive turbulence in a sample reservoir. To validate performance of this system, we present experimental evidence demonstrating prevention of blood cell settling, cell integrity, and staining of cells prior to flow cytometric analysis. This setup is further integrated with a microfluidic imaging flow cytometer to investigate cell count variability. With no need for prior sample preparation, a drop of whole blood can be directly introduced to the setup without premixing with buffers manually. Our results show that integration of this assembly with microfluidic analysis provides a competent automation tool for low-cost point-of-care blood-based diagnostics.

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.

Reticulocyte maturity index by flow cytometry: its applicability in radioinduced bone marrow aplasia

International Nuclear Information System (INIS)

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

1995-01-01

Flow cytometric reticulocyte quantification was assayed in ten patients undergoing bone marrow transplantation (BMT) with previous conditioning chemotherapy and total body irradiation (TBI). A reticulocyte maturity index (RMI) was determined taking into account the RNA content. With de aim of testing the utility of RMI as an early predictor of functional recovery in marrow aplasia, other haematological 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 following of radioinduced bone marrow aplasia is discussed. (author). 4 refs., 4 figs., 2 tabs

Flow cytometry for intracellular SPION quantification: specificity and sensitivity in comparison with spectroscopic methods

Directory of Open Access Journals (Sweden)

Friedrich RP

2015-06-01

Full Text Available Ralf P Friedrich,1 Christina Janko,1 Marina Poettler,1 Philipp Tripal,1 Jan Zaloga,1 Iwona Cicha,1 Stephan Dürr,1,2 Johannes Nowak,3 Stefan Odenbach,3 Ioana Slabu,4 Maik Liebl,4 Lutz Trahms,4 Marcus Stapf,5 Ingrid Hilger,5 Stefan Lyer,1 Christoph Alexiou1 1Department of Otorhinolaryngology, Head and Neck Surgery, Section of Experimental Oncology and Nanomedicine, University hospital Erlangen, 2Department of Otorhinolaryngology, Head and Neck Surgery, Section of Phoniatrics and Pediatric Audiology, University hospital Erlangen, Erlangen, 3Technische Universität Dresden, Chair of Magnetofluiddynamics, Measuring and Automation Technology, Dresden, 4Physikalisch-Technische Bundesanstalt Berlin, Berlin, 5Department of Radiology, Division of Diagnostic and Interventional Radiology, Experimental Radiology, University hospital Jena, Jena, Germany Abstract: Due to their special physicochemical properties, iron nanoparticles offer new promising possibilities for biomedical applications. For bench to bedside translation of superparamagnetic iron oxide nanoparticles (SPIONs, safety issues have to be comprehensively clarified. To understand concentration-dependent nanoparticle-mediated toxicity, the exact quantification of intracellular SPIONs by reliable methods is of great importance. In the present study, we compared three different SPION quantification methods (ultraviolet spectrophotometry, magnetic particle spectroscopy, atomic adsorption spectroscopy and discussed the shortcomings and advantages of each method. Moreover, we used those results to evaluate the possibility to use flow cytometric technique to determine the cellular SPION content. For this purpose, we correlated the side scatter data received from flow cytometry with the actual cellular SPION amount. We showed that flow cytometry provides a rapid and reliable method to assess the cellular SPION content. Our data also demonstrate that internalization of iron oxide nanoparticles in human

Fast Reliability Assessing Method for Distribution Network with Distributed Renewable Energy Generation

Science.gov (United States)

Chen, Fan; Huang, Shaoxiong; Ding, Jinjin; Ding, Jinjin; Gao, Bo; Xie, Yuguang; Wang, Xiaoming

2018-01-01

This paper proposes a fast reliability assessing method for distribution grid with distributed renewable energy generation. First, the Weibull distribution and the Beta distribution are used to describe the probability distribution characteristics of wind speed and solar irradiance respectively, and the models of wind farm, solar park and local load are built for reliability assessment. Then based on power system production cost simulation probability discretization and linearization power flow, a optimal power flow objected with minimum cost of conventional power generation is to be resolved. Thus a reliability assessment for distribution grid is implemented fast and accurately. The Loss Of Load Probability (LOLP) and Expected Energy Not Supplied (EENS) are selected as the reliability index, a simulation for IEEE RBTS BUS6 system in MATLAB indicates that the fast reliability assessing method calculates the reliability index much faster with the accuracy ensured when compared with Monte Carlo method.

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, P<0.05) of the variation in the conception rate and included nine variables: five kinetic parameters measured by CASA (total motility, active cells, beat cross frequency, curvilinear velocity and amplitude of lateral head displacement) and four parameters related to DNA integrity evaluated by FCM (degree of chromatin structure abnormality Alpha-T, extent of chromatin structure abnormality (Alpha-T standard deviation), percentage of DNA fragmented sperm and percentage of sperm with high green fluorescence representative of immature cells). A significant relationship (R 2=0.84, P<0.05) was observed between

Using flow cytometry for counting natural planktonic bacteria and understanding the structure of planktonic bacterial communities

Directory of Open Access Journals (Sweden)

Josep M. Gasol

2000-06-01

Full Text Available Flow cytometry is rapidly becoming a routine methodology in aquatic microbial ecology. The combination of simple to use bench-top flow cytometers and highly fluorescent nucleic acid stains allows fast and easy determination of microbe abundance in the plankton of lakes and oceans. The different dyes and protocols used to stain and count planktonic bacteria as well as the equipment in use are reviewed, with special attention to some of the problems encountered in daily routine practice such as fixation, staining and absolute counting. One of the main advantages of flow cytometry over epifluorescence microscopy is the ability to obtain cell-specific measurements in large numbers of cells with limited effort. We discuss how this characteristic has been used for differentiating photosynthetic from non-photosynthetic prokaryotes, for measuring bacterial cell size and nucleic acid content, and for estimating the relative activity and physiological state of each cell. We also describe how some of the flow cytometrically obtained data can be used to characterize the role of microbes on carbon cycling in the aquatic environment and we prospect the likely avenues of progress in the study of planktonic prokaryotes through the use of flow cytometry.

Three dimensional quantitative coronary angiography can detect reliably ischemic coronary lesions based on fractional flow reserve.

Science.gov (United States)

Chung, Woo-Young; Choi, Byoung-Joo; Lim, Seong-Hoon; Matsuo, Yoshiki; Lennon, Ryan J; Gulati, Rajiv; Sandhu, Gurpreet S; Holmes, David R; Rihal, Charanjit S; Lerman, Amir

2015-06-01

Conventional coronary angiography (CAG) has limitations in evaluating lesions producing ischemia. Three dimensional quantitative coronary angiography (3D-QCA) shows reconstructed images of CAG using computer based algorithm, the Cardio-op B system (Paieon Medical, Rosh Ha'ayin, Israel). The aim of this study was to evaluate whether 3D-QCA can reliably predict ischemia assessed by myocardial fractional flow reserve (FFR) < 0.80. 3D-QCA images were reconstructed from CAG which also were evaluated with FFR to assess ischemia. Minimal luminal diameter (MLD), percent diameter stenosis (%DS), minimal luminal area (MLA), and percent area stenosis (%AS) were obtained. The results of 3D-QCA and FFR were compared. A total of 266 patients was enrolled for the present study. FFR for all lesions ranged from 0.57 to 1.00 (0.85 ± 0.09). Measurement of MLD, %DS, MLA, and %AS all were significantly correlated with FFR (r = 0.569, 0609, 0.569, 0.670, respectively, all P < 0.001). In lesions with MLA < 4.0 mm(2), %AS of more than 65.5% had a 80% sensitivity and a 83% specificity to predict FFR < 0.80 (area under curve, AUC was 0.878). 3D-QCA can reliably predict coronary lesions producing ischemia and may be used to guide therapeutic approach for coronary artery disease.

Reliability of the grid as function of power flow and configuration during maintenance

International Nuclear Information System (INIS)

Kovacs, Z.; Hlavac, P.; Janicek, F.

2012-01-01

The paper describes using of PSA (Probabilistic Safety Assessment) for analysis of the partial and total blackout of the grid for different power flows and configuration of the grid given by the preventive maintenance activities. The method uses event and fault tree analysis in combination with the dynamic stability analysis of the grid. The event trees model the grid response to the initiating event of the accident. The fault trees are used to model the reliability of the substation protection systems. Different end states of the event trees are simulated on the basis of dynamic stability analysis, as safe and emergency state, partial and total blackout. Minimal cut sets are generated, importance measures (Fussel-Vesely importance, risk increase factor and risk decrease factors) are calculated for the total and partial blackout. The most dominant initiating events and component failures are identified from the risk point of view. Using the PSA the model, of the transmission grid is being developed. The model can be implemented into the risk monitor software to analyze the risk for different configurations of the grid and to support the operation and the maintenance activities. (Authors)

Identification of ataxia telangiectasia heterozygotes by flow cytometric analysis of X-ray damage

International Nuclear Information System (INIS)

Rudolph, N.S.

1989-01-01

Flow cytometry was used to identify heterozygotes for the autosomal recessive DNA-repair deficiency disease ataxia telangiectasia (AT). Confluent G 0 /G 1 fibroblasts from 4 homozygotes (at/at), 5 obligate heterozygates (at/+) and 7 presumed normal (+/+) were X-irradiated with 200 Rad and subcultured immediately in medium containing 5-bromodeoxyuridine (BrdU). Cells were harvested 72 h later and stained with fluoresceinated anti-BrdU antibody to identify cells that had entered S phase. They were counterstained with propidium iodide to measure total DNA content. On the basis of relative release from G 0 /G 1 , the at/+ strains as a group were distinguished from both the presumed +/+ strains and at/at strains, although the individual values for some strains did show overlap between genotypes. When 10 cell strains were coded and analyzed in 'blind' experiments, all 4 heterozygotes were correctly assigned. By a similar assay in which exponentially growing cultures were pulsed briefly with BrdU 8 h after irradiation with 400 Rad and then harvested immediately, presumed +/+ cells as a group could be distinguished from at/at cells but not from at/- cells. This combination of assays assists in the identification of all 3 AT genotypes. This should be of both basic and diagnostic use, particularly in families known to segregate AT. (author). 37 refs.; 3 figs.; 5 tabs

Measurement of lipid accumulation in Chlorella vulgaris via flow cytometry and liquid-state ¹H NMR spectroscopy for development of an NMR-traceable flow cytometry protocol.

Directory of Open Access Journals (Sweden)

Michael S Bono

Full Text Available In this study, we cultured Chlorella vulgaris cells with a range of lipid contents, induced via nitrogen starvation, and characterized them via flow cytometry, with BODIPY 505/515 as a fluorescent lipid label, and liquid-state 1H NMR spectroscopy. In doing so, we demonstrate the utility of calibrating flow cytometric measurements of algal lipid content using triacylglyceride (TAG, also known as triacylglycerol or triglyceride content per cell as measured via quantitative 1H NMR. Ensemble-averaged fluorescence of BODIPY-labeled cells was highly correlated with average TAG content per cell measured by bulk NMR, with a linear regression yielding a linear fit with r2 = 0.9974. This correlation compares favorably to previous calibrations of flow cytometry protocols to lipid content measured via extraction, and calibration by NMR avoids the time and complexity that is generally required for lipid quantitation via extraction. Flow cytometry calibrated to a direct measurement of TAG content can be used to investigate the distribution of lipid contents for cells within a culture. Our flow cytometry measurements showed that Chlorella vulgaris cells subjected to nitrogen limitation exhibited higher mean lipid content but a wider distribution of lipid content that overlapped the relatively narrow distribution of lipid content for replete cells, suggesting that nitrogen limitation induces lipid accumulation in only a subset of cells. Calibration of flow cytometry protocols using direct in situ measurement of TAG content via NMR will facilitate rapid development of more precise flow cytometry protocols, enabling investigation of algal lipid accumulation for development of more productive algal biofuel feedstocks and cultivation protocols.

Measurement of Lipid Accumulation in Chlorella vulgaris via Flow Cytometry and Liquid-State ¹H NMR Spectroscopy for Development of an NMR-Traceable Flow Cytometry Protocol

Science.gov (United States)

Bono Jr., Michael S.; Garcia, Ravi D.; Sri-Jayantha, Dylan V.; Ahner, Beth A.; Kirby, Brian J.

2015-01-01

In this study, we cultured Chlorella vulgaris cells with a range of lipid contents, induced via nitrogen starvation, and characterized them via flow cytometry, with BODIPY 505/515 as a fluorescent lipid label, and liquid-state 1H NMR spectroscopy. In doing so, we demonstrate the utility of calibrating flow cytometric measurements of algal lipid content using triacylglyceride (TAG, also known as triacylglycerol or triglyceride) content per cell as measured via quantitative 1H NMR. Ensemble-averaged fluorescence of BODIPY-labeled cells was highly correlated with average TAG content per cell measured by bulk NMR, with a linear regression yielding a linear fit with r 2 = 0.9974. This correlation compares favorably to previous calibrations of flow cytometry protocols to lipid content measured via extraction, and calibration by NMR avoids the time and complexity that is generally required for lipid quantitation via extraction. Flow cytometry calibrated to a direct measurement of TAG content can be used to investigate the distribution of lipid contents for cells within a culture. Our flow cytometry measurements showed that Chlorella vulgaris cells subjected to nitrogen limitation exhibited higher mean lipid content but a wider distribution of lipid content that overlapped the relatively narrow distribution of lipid content for replete cells, suggesting that nitrogen limitation induces lipid accumulation in only a subset of cells. Calibration of flow cytometry protocols using direct in situ measurement of TAG content via NMR will facilitate rapid development of more precise flow cytometry protocols, enabling investigation of algal lipid accumulation for development of more productive algal biofuel feedstocks and cultivation protocols. PMID:26267664

Measurement of lipid accumulation in Chlorella vulgaris via flow cytometry and liquid-state ¹H NMR spectroscopy for development of an NMR-traceable flow cytometry protocol.

Science.gov (United States)

Bono, Michael S; Garcia, Ravi D; Sri-Jayantha, Dylan V; Ahner, Beth A; Kirby, Brian J

2015-01-01

In this study, we cultured Chlorella vulgaris cells with a range of lipid contents, induced via nitrogen starvation, and characterized them via flow cytometry, with BODIPY 505/515 as a fluorescent lipid label, and liquid-state 1H NMR spectroscopy. In doing so, we demonstrate the utility of calibrating flow cytometric measurements of algal lipid content using triacylglyceride (TAG, also known as triacylglycerol or triglyceride) content per cell as measured via quantitative 1H NMR. Ensemble-averaged fluorescence of BODIPY-labeled cells was highly correlated with average TAG content per cell measured by bulk NMR, with a linear regression yielding a linear fit with r2 = 0.9974. This correlation compares favorably to previous calibrations of flow cytometry protocols to lipid content measured via extraction, and calibration by NMR avoids the time and complexity that is generally required for lipid quantitation via extraction. Flow cytometry calibrated to a direct measurement of TAG content can be used to investigate the distribution of lipid contents for cells within a culture. Our flow cytometry measurements showed that Chlorella vulgaris cells subjected to nitrogen limitation exhibited higher mean lipid content but a wider distribution of lipid content that overlapped the relatively narrow distribution of lipid content for replete cells, suggesting that nitrogen limitation induces lipid accumulation in only a subset of cells. Calibration of flow cytometry protocols using direct in situ measurement of TAG content via NMR will facilitate rapid development of more precise flow cytometry protocols, enabling investigation of algal lipid accumulation for development of more productive algal biofuel feedstocks and cultivation protocols.

Increase of hydroelectric power plant operation reliability

International Nuclear Information System (INIS)

Koshumbaev, M.B.

2006-01-01

The new design of the turbine of hydroelectric power plant (HPP) is executed in the form of a pipe with plates. Proposed solution allows increasing the hydroelectric power plant capacity at existing head and water flow. At that time the HPP turbine reliability is increase, its operation performances are improving. Design efficiency is effective mostly for small-scale and micro-HPP due to reliable operation, low-end technology, and harmless ecological application. (author)

Monitoring of Legionella pneumophila viability after chlorine dioxide treatment using flow cytometry.

Science.gov (United States)

Mustapha, Pascale; Epalle, Thibaut; Allegra, Séverine; Girardot, Françoise; Garraud, Olivier; Riffard, Serge

2015-04-01

The viability of three Legionella pneumophila strains was monitored after chlorine dioxide (ClO2) treatment using a flow cytometric assay. Suspensions of L. pneumophila cells were submitted to increasing concentrations of ClO2. Culturable cells were still detected when using 4 mg/L, but could no longer be detected after exposure to 6 mg/L of ClO2, although viable but not culturable (VBNC) cells were found after exposure to 4-5 mg/L of ClO2. When testing whether these VBNC were infective, two of the strains were resuscitated after co-culture with Acanthamoeba polyphaga, but neither of them could infect macrophage-like cells. Copyright © 2015 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

faloabi@uniben.edu Antiproliferative and Pro-apoptotic activities

African Journals Online (AJOL)

MICHAEL HORSFALL

Keyword: Persea americana, antiproliferative activity, apoptotic effect, flow ... of the stem bark of Persea americana in MCF-7 cell line by flow cytometer. .... of an electric milling machine. ... Flow Cytometric Measurement Of Cell Proliferation:.

A heuristic-based approach for reliability importance assessment of energy producers

International Nuclear Information System (INIS)

Akhavein, A.; Fotuhi Firuzabad, M.

2011-01-01

Reliability of energy supply is one of the most important issues of service quality. On one hand, customers usually have different expectations for service reliability and price. On the other hand, providing different level of reliability at load points is a challenge for system operators. In order to take reasonable decisions and obviate reliability implementation difficulties, market players need to know impacts of their assets on system and load-point reliabilities. One tool to specify reliability impacts of assets is the criticality or reliability importance measure by which system components can be ranked based on their effect on reliability. Conventional methods for determination of reliability importance are essentially on the basis of risk sensitivity analysis and hence, impose prohibitive calculation burden in large power systems. An approach is proposed in this paper to determine reliability importance of energy producers from perspective of consumers or distribution companies in a composite generation and transmission system. In the presented method, while avoiding immense computational burden, the energy producers are ranked based on their rating, unavailability and impact on power flows in the lines connecting to the considered load points. Study results on the IEEE reliability test system show successful application of the proposed method. - Research highlights: → Required reliability level at load points is a concern in modern power systems. → It is important to assess reliability importance of energy producers or generators. → Generators can be ranked based on their impacts on power flow to a selected area. → Ranking of generators is an efficient tool to assess their reliability importance.

An evaluation of the multi-state node networks reliability using the traditional binary-state networks reliability algorithm

International Nuclear Information System (INIS)

Yeh, W.-C.

2003-01-01

A system where the components and system itself are allowed to have a number of performance levels is called the Multi-state system (MSS). A multi-state node network (MNN) is a generalization of the MSS without satisfying the flow conservation law. Evaluating the MNN reliability arises at the design and exploitation stage of many types of technical systems. Up to now, the known existing methods can only evaluate a special MNN reliability called the multi-state node acyclic network (MNAN) in which no cyclic is allowed. However, no method exists for evaluating the general MNN reliability. The main purpose of this article is to show first that each MNN reliability can be solved using any the traditional binary-state networks (TBSN) reliability algorithm with a special code for the state probability. A simple heuristic SDP algorithm based on minimal cuts (MC) for estimating the MNN reliability is presented as an example to show how the TBSN reliability algorithm is revised to solve the MNN reliability problem. To the author's knowledge, this study is the first to discuss the relationships between MNN and TBSN and also the first to present methods to solve the exact and approximated MNN reliability. One example is illustrated to show how the exact MNN reliability is obtained using the proposed algorithm

Flow cytometric analysis of lymphocyte subset in patients with neutropenia among atomic bomb survivors

Energy Technology Data Exchange (ETDEWEB)

Imamura, Nobutaka; Kimura, Akiro [Hiroshima Univ. (Japan). Research Inst. for Radiation Biology and Medicine

1998-12-01

In 51 patients (atomic bomb survivors 50, unexposed persons 1) who have had neutropenia for two years or more under indistinct cause, cell surface antigen was analyzed by flow cytometry. Twenty-nine cases of survivors were diagnosed as NK cell leukemia or NK cell cytosis because analysis data showed CD3(-), CD56(+) and CD57(+/-). Six cases were diagnosed as NK like T cell hypercytosis because analysis data showed CD3(+), CD56(+/-) and CD57(+). As for 15 cases, CD56(+) cell number was in range of 15.96{+-}5.35 of a normal person, and no relation with NK cell was recognized. But, CD4/CD8 ratio was higher than 2.1, and gain of T helper cell was recognized. One unexposed persons was diagnosed as chronic NK cell leukemia because analysis data showed CD3(-), CD56(+) and CD57(+). Anti-neutrophil antibody wasn't recognized. Cytotoxic activity for K562 and Raji cell line showed high value compared with that of a normal person. Epstein Barr virus wasn't detected. (K.H.)

Chromosome specific DNA hybridization in suspension for flow cytometric detection of chimerism in bone marrow transplantation and leukemia

NARCIS (Netherlands)

G.J.A. Arkesteijn (Ger); C.A.J. Erpelinck (Claudia); A.C.M. Martens (Anton); A. Hagenbeek (Anton)

1995-01-01

textabstractFlow cytometry was used to measure the fluorescence intensity of nuclei that were subjected to fluorescent in situ hybridization in suspension with chromosome specific DNA probes. Paraformaldehyde-fixed nuclei were protein digested with trypsin and hybridized simultaneously with a

Physiological Studies of Lactococcus lactis

DEFF Research Database (Denmark)

Hansen, Gunda

medium and (specific) acidification activity in milk in response to the extracellular pH (pHex) during batch fermentations and in response to stress during downstream processing and storage as frozen, freeze- and spray-dried cells. In this PhD thesis, in situ flow cytometric viability assessment...... fermentation, downstream processing and storage in the absence of a protectant. However, storing freeze-dried L. lactis cells at 30 °C negatively affected the culturability and acidification activity. The reactivation of freeze-dried cells in fermentation medium prior to flow cytometric viability assessment...

Analysis and assessment of water treatment plant reliability

Directory of Open Access Journals (Sweden)

Szpak Dawid

2017-03-01

Full Text Available The subject of the publication is the analysis and assessment of the reliability of the surface water treatment plant (WTP. In the study the one parameter method of reliability assessment was used. Based on the flow sheet derived from the water company the reliability scheme of the analysed WTP was prepared. On the basis of the daily WTP work report the availability index Kg for the individual elements included in the WTP, was determined. Then, based on the developed reliability scheme showing the interrelationships between elements, the availability index Kg for the whole WTP was determined. The obtained value of the availability index Kg was compared with the criteria values.

Flow cytometric analysis of cell killing by the jumper ant venom peptide pilosulin 1.

Science.gov (United States)

King, M A; Wu, Q X; Donovan, G R; Baldo, B A

1998-08-01

Pilosulin 1 is a synthetic 56-amino acid residue polypeptide that corresponds to the largest allergenic polypeptide found in the venom of the jumper ant Myrmecia pilosula. Initial experiments showed that pilosulin 1 lysed erythrocytes and killed proliferating B cells. Herein, we describe how flow cytometry was used to investigate the cytotoxicity of the peptide for human white blood cells. Cells were labeled with fluorochrome-conjugated antibodies, incubated with the peptide and 7-aminoactinomycin D (7-AAD), and then analyzed. The effects of varying the peptide concentration, serum concentration, incubation time, and incubation temperature were measured, and the cytotoxicity of pilosulin 1 was compared with that of the bee venom peptide melittin. The antibodies and the 7-AAD enabled the identification of cell subpopulations and dead cells, respectively. It was possible, using the appropriate mix of antibodies and four-color analysis, to monitor the killing of three or more cell subpopulations simultaneously. We found that 1) pilosulin 1 killed cells within minutes, with kinetics similar to those of melittin; 2) pilosulin 1 was a slightly more potent cytotoxic agent than melittin; 3) both pilosulin 1 and melittin were more potent against mononuclear leukocytes than against granulocytes; and 4) serum inhibited killing by either peptide.

The use of flow cytometry to examine calcium signalling by TRPV1 in mixed cell populations.

Science.gov (United States)

Assas, Bakri M; Abdulaal, Wesam H; Wakid, Majed H; Zakai, Haytham A; Miyan, J; Pennock, J L

2017-06-15

Flow cytometric analysis of calcium mobilisation has been in use for many years in the study of specific receptor engagement or isolated cell:cell communication. However, calcium mobilisation/signaling is key to many cell functions including apoptosis, mobility and immune responses. Here we combine multiplex surface staining of whole spleen with Indo-1 AM to visualise calcium mobilisation and examine calcium signaling in a mixed immune cell culture over time. We demonstrate responses to a TRPV1 agonist in distinct cell subtypes without the need for cell separation. Multi parameter staining alongside Indo-1 AM to demonstrate calcium mobilization allows the study of real time calcium signaling in a complex environment. Copyright © 2017. Published by Elsevier Inc.

Effect of flow conditions on flow accelerated corrosion in pipe bends

International Nuclear Information System (INIS)

Mazhar, H.; Ching, C.Y.

2015-01-01

Flow Accelerated Corrosion (FAC) in piping systems is a safety and reliability problem in the nuclear industry. In this study, the pipe wall thinning rates and development of surface roughness in pipe bends are compared for single phase and two phase annular flow conditions. The FAC rates were measured using the dissolution of test sections cast from gypsum in water with a Schmidt number of 1280. The change in location and levels of maximum FAC under single phase and two phase flow conditions are examined. The comparison of the relative roughness indicates a higher effect for the surface roughness in single phase flow than in two phase flow. (author)

Use of Melt Flow Rate Test in Reliability Study of Thermoplastic Encapsulation Materials in Photovoltaic Modules

Energy Technology Data Exchange (ETDEWEB)

Moseley, J.; Miller, D.; Shah, Q.-U.-A. S. J.; Sakurai, K.; Kempe, M.; Tamizhmani, G.; Kurtz, S.

2011-10-01

Use of thermoplastic materials as encapsulants in photovoltaic (PV) modules presents a potential concern in terms of high temperature creep, which should be evaluated before thermoplastics are qualified for use in the field. Historically, the issue of creep has been avoided by using thermosetting polymers as encapsulants, such as crosslinked ethylene-co-vinyl acetate (EVA). Because they lack crosslinked networks, however, thermoplastics may be subject to phase transitions and visco-elastic flow at the temperatures and mechanical stresses encountered by modules in the field, creating the potential for a number of reliability and safety issues. Thermoplastic materials investigated in this study include PV-grade uncured-EVA (without curing agents and therefore not crosslinked); polyvinyl butyral (PVB); thermoplastic polyurethane (TPU); and three polyolefins (PO), which have been proposed for use as PV encapsulation. Two approaches were used to evaluate the performance of these materials as encapsulants: module-level testing and a material-level testing.

Flow-cytometric analysis of mouse embryonic stem cell lipofection using small and large DNA constructs.

Science.gov (United States)

McLenachan, Samuel; Sarsero, Joseph P; Ioannou, Panos A

2007-06-01

Using the lipofection reagent LipofectAMINE 2000 we have examined the delivery of plasmid DNA (5-200 kb) to mouse embryonic stem (mES) cells by flow cytometry. To follow the physical uptake of lipoplexes we labeled DNA molecules with the fluorescent dye TOTO-1. In parallel, expression of an EGFP reporter cassette in constructs of different sizes was used as a measure of nuclear delivery. The cellular uptake of DNA lipoplexes is dependent on the uptake competence of mES cells, but it is largely independent of DNA size. In contrast, nuclear delivery was reduced with increasing plasmid size. In addition, linear DNA is transfected with lower efficiency than circular DNA. Inefficient cytoplasmic trafficking appears to be the main limitation in the nonviral delivery of large DNA constructs to the nucleus of mES cells. Overcoming this limitation should greatly facilitate functional studies with large genomic fragments in embryonic stem cells.

Finding Elephant Flows for Optical Networks

NARCIS (Netherlands)

Fioreze, Tiago; Oude Wolbers, Mattijs; van de Meent, R.; Pras, Aiko

2007-01-01

Optical networks are fast and reliable networks that enable, amongst others, dedicated light paths to be established for elephant IP flows. Elephant IP flows are characterized by being small in number, but long in time and high in traffic volume. Moving these flows from the general IP network to

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

14CO2 labeling: a reliable technique for rapid measurement of total root exudation capacity and vascular sap flow in crops

International Nuclear Information System (INIS)

Singh, Bhupinder; Ahuja, Sumedha; Pandey, Renu; Singhal, R.K.

2014-01-01

Ability of roots to release organic compounds in its rhizosphere is known to improve plant available nutrients and reduces heavy metal toxicity by immobilization. It is regarded as an important determinant of micro nutrient deficiency tolerance in plants. Uptake of nutrients and translocation of photoassimilates, on the other hand are governed by the strength of the transpiration stream and sink demand respectively. Measurement of vascular sap flow, thus, is critical for understanding of the translocation efficiency and consequently the sink demand that keeps changing during the crop growth cycle. Measurement of the root exudation capacity and the vascular sap flow is cumbersome and time consuming. Since, the exudates released by the roots and the photosynthates translocated between the source and the sink are essentially carbon compounds, use of labeled carbon as tag could potentially be exploited for a rapid and reliable measurement of exudation and vascular sap flow in crop plants. We report here the experimental results involving 14 C labeling of groundnut, a legume crop, as 14 CO 2 generated by acidification of sodium bicarbonate. An additional factor of seed gamma irradiation was used to generate variability in the root exudation and the sap flow. The 14 C release by the roots was compared against the 14 C transport in the vascular sap. An experimental hypothesis that a higher 14 C level in the vascular sap would indicate a higher root release of carbon by the roots into the rhizosphere was verified. (author)

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.

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

Flow cytometric characterization of phenotype, DNA indices and p53 gene expression in 55 cases of acute leukemia.

Science.gov (United States)

Powari, Manish; Varma, Neelam; Varma, Subhash; Marwaha, Ram Kumar; Sandhu, Harpreet; Ganguly, Nirmal Kumar

2002-06-01

To characterize the phenotype of acute leukemia cases using flow cytometry, to detect mixed lineage cases and to use DNA index determination, including S-phase fraction (SPF) and p53 detection, to find if there was any correlation of SPF and p53 expression with outcome. Fifty-five cases of acute leukemia were enrolled in this study. A complete hemogram and routine bone marrow examination, including cytochemistry, was done. Mycloperoxidase-negative cases were evaluated on a flow cytometer using monoclonal antibodies. DNA indices were determined by flow cytometry in all cases, and p53 was detected immunohistochemically using the alkaline phosphatase/antialkaline phosphatase technique. Acute myeloblastic leukemia (AML) was diagnosed in 32 cases; acute lymphoblastic leukemia (ALL) was diagnosed in 18 (14 B lineage and 4 T line age). Four cases showed mixed lineage leukemia, and undifferentiated acute leukemia was diagnosed in one case. The mean/range of SPF for these groups were 3.76/0.33-6.91, 6.25/0.15-21.4, 2.89/0.35-10.64, 2.60/0.72-6.94 and 7.34, respectively. Aneuploidy was detected in two cases of B-lineage ALL and tetraploidy in a case of AML-M7, while all others were diploid p53. Was detected in 6 of 55 cases (10.90%). Follow-up was available for 24 patients. Five patients relapsed, and four had B-cell type ALL and were diploid and expressed no p53 gene. SPF% did not show any correlation with outcome. These data suggest that within acute leukemia subtypes, there is a wide variation in SPF. SPF does not seem to correlate with outcome. Immunophenotyping is essential to determine the lineage in myeloperoxidase-negative cases. It is perhaps the only way to diagnose mixed lineage leukemia and aberrant expression of markers presently. The p53 gene was detected less frequently. However, more studies are required from different centers with longer follow-up to evaluate prognostic significance.

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, ...

Validity and reliability of a novel slow cuff-deflation system for noninvasive blood pressure monitoring in patients with continuous-flow left ventricular assist device.

Science.gov (United States)

Lanier, Gregg M; Orlanes, Khristine; Hayashi, Yacki; Murphy, Jennifer; Flannery, Margaret; Te-Frey, Rosie; Uriel, Nir; Yuzefpolskaya, Melana; Mancini, Donna M; Naka, Yoshifumi; Takayama, Hiroo; Jorde, Ulrich P; Demmer, Ryan T; Colombo, Paolo C

2013-09-01

Doppler ultrasound is the clinical gold standard for noninvasive blood pressure (BP) measurement among continuous-flow left ventricular assist device patients. The relationship of Doppler BP to systolic BP (SBP) and mean arterial pressure (MAP) is uncertain and Doppler measurements require a clinic visit. We studied the relationship between Doppler BP and both arterial-line (A-line) SBP and MAP. Validity and reliability of the Terumo Elemano BP Monitor, a novel slow cuff-deflation device that could potentially be used by patients at home, were assessed. Doppler and Terumo BP measurements were made in triplicate among 60 axial continuous-flow left ventricular assist device (HeartMate II) patients (30 inpatients and 30 outpatients) at 2 separate exams (360 possible measurements). A-line measures were also obtained among inpatients. Mean absolute differences (MADs) and correlations were used to determine within-device reliability (comparison of second and third BP measures) and between-device validity. Bland-Altman plots assessed BP agreement between A-line, Doppler BP, and Terumo Elemano. Success rates for Doppler and Terumo Elemano were 100% and 91%. Terumo Elemano MAD for repeat SBP and MAP were 4.6±0.6 and 4.2±0.6 mm Hg; repeat Doppler BP MAD was 2.9±0.2 mm Hg. Mean Doppler BP was lower than A-line SBP by 4.1 (MAD=6.4±1.4) mm Hg and higher than MAP by 9.5 (MAD=11.0±1.2) mm Hg; Terumo Elemano underestimated A-line SBP by 0.3 (MAD=5.6±0.9) mm Hg and MAP by 1.7 (MAD=6.0±1.0) mm Hg. Doppler BP more closely approximates SBP than MAP. Terumo Elemano was successful, reliable, and valid when compared with A-line and Doppler.

Evaluate the system reliability for a manufacturing network with reworking actions

International Nuclear Information System (INIS)

Lin, Yi-Kuei; Chang, Ping-Chen

2012-01-01

To measure the system reliability of a manufacturing system with reworking actions is a crucial issue in industry, in which the system reliability could be one of the essential performance indicators to evaluate whether the manufacturing system is capable or not. In a manufacturing system, the input flow (raw materials/WIP) processed by each machine might be defective and thus the output flow (WIP/products) would be less than the input amount. Moreover, defective WIP/products are usually incentive to be reworked for reducing wasting and increasing output. Therefore, reworking actions are necessary to be considered in the manufacturing system. Based on the path concept, we revise such a manufacturing system as a stochastic-flow network in which the capacity of each machine is stochastic (i.e., multistate) due to the failure, partial failure, and maintenance. We decompose the network into one general processing path and several reworking paths. Subsequently, three algorithms for different network models are proposed to generate the lower boundary vector which affords to produce enough products satisfying the demand d. In terms of such a vector, the system reliability can be derived afterwards.

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...

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

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...

77 FR 39858 - Revisions to Electric Reliability Organization Definition of Bulk Electric System and Rules of...

Science.gov (United States)

2012-07-05

... bulk electric system reliability through steady state power flow, and contain a transient stability... Commission 18 CFR Part 40 Revisions to Electric Reliability Organization Definition of Bulk Electric System... definition of ``bulk electric system'' developed by the North American Electric Reliability Corporation (NERC...

Functional characterization of neotropical snakes peripheral blood leukocytes subsets: Linking flow cytometry cell features, microscopy images and serum corticosterone levels.

Science.gov (United States)

de Carvalho, Marcelo Pires Nogueira; Queiroz-Hazarbassanov, Nicolle Gilda Teixeira; de Oliveira Massoco, Cristina; Sant'Anna, Sávio Stefanini; Lourenço, Mariana Mathias; Levin, Gabriel; Sogayar, Mari Cleide; Grego, Kathleen Fernandes; Catão-Dias, José Luiz

2017-09-01

Reptiles are the unique ectothermic amniotes, providing the key link between ectothermic anamniotes fish and amphibians, and endothermic birds and mammals; becoming an important group to study with the aim of providing significant knowledge into the evolutionary history of vertebrate immunity. Classification systems for reptiles' leukocytes have been described by their appearance rather than function, being still inconsistent. With the advent of modern techniques and the establishment of analytical protocols for snakes' blood by flow cytometry, we bring a qualitative and quantitative assessment of innate activities presented by snakes' peripheral blood leukocytes, thereby linking flow cytometric features with fluorescent and light microscopy images. Moreover, since corticosterone is an important immunomodulator in reptiles, hormone levels of all blood samples were measured. We provide novel and additional information which should contribute to better understanding of the development of the immune system of reptiles and vertebrates. Copyright © 2017 Elsevier Ltd. All rights reserved.

Ternary networks reliability and Monte Carlo

CERN Document Server

Gertsbakh, Ilya; Vaisman, Radislav

2014-01-01

Ternary means “based on three”. This book deals with reliability investigations of  networks whose components subject to failures can be in three states –up, down and middle (mid), contrary to traditionally considered networks  having only binary (up/down) components. Extending binary case to ternary allows to consider more realistic and flexible models for communication, flow and supply networks.

ICCS/ESCCA consensus guidelines to detect GPI-deficient cells in paroxysmal nocturnal hemoglobinuria (PNH) and related disorders part 4 - assay validation and quality assurance.

Science.gov (United States)

Oldaker, Teri; Whitby, Liam; Saber, Maryam; Holden, Jeannine; Wallace, Paul K; Litwin, Virginia

2018-01-01

Over the past six years, a diverse group of stakeholders have put forth recommendations regarding the analytical validation of flow cytometric methods and described in detail the differences between cell-based and traditional soluble analyte assay validations. This manuscript is based on these general recommendations as well as the published experience of experts in the area of PNH testing. The goal is to provide practical assay-specific guidelines for the validation of high-sensitivity flow cytometric PNH assays. Examples of the reports and validation data described herein are provided in Supporting Information. © 2017 International Clinical Cytometry Society. © 2017 International Clinical Cytometry Society.

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

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.)

A Real-Coded Genetic Algorithm with System Reduction and Restoration for Rapid and Reliable Power Flow Solution of Power Systems

Directory of Open Access Journals (Sweden)

Hassan Abdullah Kubba

2015-05-01

Full Text Available The paper presents a highly accurate power flow solution, reducing the possibility of ending at local minima, by using Real-Coded Genetic Algorithm (RCGA with system reduction and restoration. The proposed method (RCGA is modified to reduce the total computing time by reducing the system in size to that of the generator buses, which, for any realistic system, will be smaller in number, and the load buses are eliminated. Then solving the power flow problem for the generator buses only by real-coded GA to calculate the voltage phase angles, whereas the voltage magnitudes are specified resulted in reduced computation time for the solution. Then the system is restored by calculating the voltages of the load buses in terms of the calculated voltages of the generator buses, after a derivation of equations for calculating the voltages of the load busbars. The proposed method was demonstrated on 14-bus IEEE test systems and the practical system 362-busbar IRAQI NATIONAL GRID (ING. The proposed method has reliable convergence, a highly accurate solution and less computing time for on-line applications. The method can conveniently be applied for on-line analysis and planning studies of large power systems.

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.

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

Directory of Open Access Journals (Sweden)

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.

NPTool: Towards Scalability and Reliability of Business Process Management

Science.gov (United States)

Braghetto, Kelly Rosa; Ferreira, João Eduardo; Pu, Calton

Currently one important challenge in business process management is provide at the same time scalability and reliability of business process executions. This difficulty becomes more accentuated when the execution control assumes complex countless business processes. This work presents NavigationPlanTool (NPTool), a tool to control the execution of business processes. NPTool is supported by Navigation Plan Definition Language (NPDL), a language for business processes specification that uses process algebra as formal foundation. NPTool implements the NPDL language as a SQL extension. The main contribution of this paper is a description of the NPTool showing how the process algebra features combined with a relational database model can be used to provide a scalable and reliable control in the execution of business processes. The next steps of NPTool include reuse of control-flow patterns and support to data flow management.

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)

[Flow cytometric test using eosin-5'-maleimide (EMA) labelling of red blood for diagnosis of hereditary spherocytosis].

Science.gov (United States)

Wang, Jiying; Zheng, Bin; Zhao, Yuping; Chen, Xuejing; Liu, Yan; Bo, Lijin; Zheng, Yizhou; Zhang, Fengkui; Ru, Kun; Wang, Huijun

2015-07-01

To investigate the sensitivity and specificity of eosin-5'-maleimide (EMA)assay for the diagnosis of hereditary spherocytosis (HS), and to verify the stability of reagent and samples. EMA flow cytometry test, NaCl-osmotic fragility test and acidified glycerol lysis test were performed using peripheral blood samples from 80 patients with HS and 44 patients with other blood diseases, the sensitivity and specificity of the three methods were compared, and the feasibility of EMA binding test was estimated. The stability of EMA reagent and HS samples stored at different temperatures were tested. Among the 124 tested samples, the sensitivity and specificity of EMA binding test was 0.925 and 0.954, that of NaCl-osmotic fragility test was 0.950 and 0.455, and that of acidified glycerol lysis test was 1.000 and 0.318, respectively. Although the sensitivity of NaCl-osmotic fragility test and acidified glycerol lysis test was a little higher than that of EMA binding test, the specificity of the former two methods was poor, they couldn't clearly distinguish whether spherocytosis is hereditary spherocytosis. The experiment results showed that EMA was sensitive to the temperature and should not be stored in a small aliquots at -80 ℃ over a period of 6 months. The stability of the HS sample was better, 6 days storage at 4 ℃ and 3 days storage at room temperature had no influence on the results. EMA binding test by flow cytometry showed good sensitivity and specificity for HS diagnosis. EMA reagent should be stored at-80 ℃ and the HS samples should be tested within 6 days storage at 4 ℃ and 3 days at room temperature.

Intra-observer reliability and agreement of manual and digital orthodontic model analysis.

Science.gov (United States)

Koretsi, Vasiliki; Tingelhoff, Linda; Proff, Peter; Kirschneck, Christian

2018-01-23

Digital orthodontic model analysis is gaining acceptance in orthodontics, but its reliability is dependent on the digitalisation hardware and software used. We thus investigated intra-observer reliability and agreement / conformity of a particular digital model analysis work-flow in relation to traditional manual plaster model analysis. Forty-eight plaster casts of the upper/lower dentition were collected. Virtual models were obtained with orthoX®scan (Dentaurum) and analysed with ivoris®analyze3D (Computer konkret). Manual model analyses were done with a dial caliper (0.1 mm). Common parameters were measured on each plaster cast and its virtual counterpart five times each by an experienced observer. We assessed intra-observer reliability within method (ICC), agreement/conformity between methods (Bland-Altman analyses and Lin's concordance correlation), and changing bias (regression analyses). Intra-observer reliability was substantial within each method (ICC ≥ 0.7), except for five manual outcomes (12.8 per cent). Bias between methods was statistically significant, but less than 0.5 mm for 87.2 per cent of the outcomes. In general, larger tooth sizes were measured digitally. Total difference maxilla and mandible had wide limits of agreement (-3.25/6.15 and -2.31/4.57 mm), but bias between methods was mostly smaller than intra-observer variation within each method with substantial conformity of manual and digital measurements in general. No changing bias was detected. Although both work-flows were reliable, the investigated digital work-flow proved to be more reliable and yielded on average larger tooth sizes. Averaged differences between methods were within 0.5 mm for directly measured outcomes but wide ranges are expected for some computed space parameters due to cumulative error. © The Author 2017. Published by Oxford University Press on behalf of the European Orthodontic Society. All rights reserved. For permissions, please email: journals.permissions@oup.com

Mercury flow experiments. 4th report Measurements of erosion rate caused by mercury flow

CERN Document Server

Kinoshita, H; Hino, R; Kaminaga, M

2002-01-01

The Japan Atomic Energy Research Institute (JAERI) and the High Energy Accelerator Research Organization (KEK) are promoting a construction plan of the Material-Life Science Facility, which is consisted of a Muon Science Facility and a Neutron Scattering Facility, in order to open up the new science fields. The Neutron Scattering Facility will be utilized for advanced fields of Material and Life science using high intensity neutron generated by the spallation reaction of a 1 MW pulsed proton beam and mercury target. Design of the spallation mercury target system aims to obtain high neutron performance with high reliability and safety. Since the target system is using mercury as the target material and contains large amount of radioactive spallation products, it is necessary to estimate reliability for strength of instruments in a mercury flow system during lifetime of the facility. Piping and components in the mercury flow system would be damaged by erosion with mercury flow, since these components will be we...

Is our Ground-Truth for Traffic Classification Reliable?

DEFF Research Database (Denmark)

Carela-Español, Valentín; Bujlow, Tomasz; Barlet-Ros, Pere

2014-01-01

. In order to evaluate these tools we have carefully built a labeled dataset of more than 500 000 flows, which contains traffic from popular applications. Our results present PACE, a commercial tool, as the most reliable solution for ground-truth generation. However, among the open-source tools available...

Reliability Evaluation of Concentric Butterfly Valve Using Statistical Hypothesis Test

Energy Technology Data Exchange (ETDEWEB)

Chang, Mu Seong; Choi, Jong Sik; Choi, Byung Oh; Kim, Do Sik [Korea Institute of Machinery and Materials, Daejeon (Korea, Republic of)

2015-12-15

A butterfly valve is a type of flow-control device typically used to regulate a fluid flow. This paper presents an estimation of the shape parameter of the Weibull distribution, characteristic life, and B10 life for a concentric butterfly valve based on a statistical analysis of the reliability test data taken before and after the valve improvement. The difference in the shape and scale parameters between the existing and improved valves is reviewed using a statistical hypothesis test. The test results indicate that the shape parameter of the improved valve is similar to that of the existing valve, and that the scale parameter of the improved valve is found to have increased. These analysis results are particularly useful for a reliability qualification test and the determination of the service life cycles.

Reliability Evaluation of Concentric Butterfly Valve Using Statistical Hypothesis Test

International Nuclear Information System (INIS)

Chang, Mu Seong; Choi, Jong Sik; Choi, Byung Oh; Kim, Do Sik

2015-01-01

A butterfly valve is a type of flow-control device typically used to regulate a fluid flow. This paper presents an estimation of the shape parameter of the Weibull distribution, characteristic life, and B10 life for a concentric butterfly valve based on a statistical analysis of the reliability test data taken before and after the valve improvement. The difference in the shape and scale parameters between the existing and improved valves is reviewed using a statistical hypothesis test. The test results indicate that the shape parameter of the improved valve is similar to that of the existing valve, and that the scale parameter of the improved valve is found to have increased. These analysis results are particularly useful for a reliability qualification test and the determination of the service life cycles

FLOCK cluster analysis of mast cell event clustering by high-sensitivity flow cytometry predicts systemic mastocytosis.

Science.gov (United States)

Dorfman, David M; LaPlante, Charlotte D; Pozdnyakova, Olga; Li, Betty

2015-11-01

In our high-sensitivity flow cytometric approach for systemic mastocytosis (SM), we identified mast cell event clustering as a new diagnostic criterion for the disease. To objectively characterize mast cell gated event distributions, we performed cluster analysis using FLOCK, a computational approach to identify cell subsets in multidimensional flow cytometry data in an unbiased, automated fashion. FLOCK identified discrete mast cell populations in most cases of SM (56/75 [75%]) but only a minority of non-SM cases (17/124 [14%]). FLOCK-identified mast cell populations accounted for 2.46% of total cells on average in SM cases and 0.09% of total cells on average in non-SM cases (P < .0001) and were predictive of SM, with a sensitivity of 75%, a specificity of 86%, a positive predictive value of 76%, and a negative predictive value of 85%. FLOCK analysis provides useful diagnostic information for evaluating patients with suspected SM, and may be useful for the analysis of other hematopoietic neoplasms. Copyright© by the American Society for Clinical Pathology.

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

Characterization of circulating microparticles (MPs) is usually performed by flow cytometry. Annexin V, a protein that Ca(2+)-dependently binds to phosphatidylserine, has been used to define entire microparticle (MP) populations, but not all MPs bind AnxV. Recent reports have correlated Anx...... for comprehensive assessment of circulating MPs directly from platelet-poor plasma with characterization of AnxV-binding and of cellular origin of MPs....

Stability of eosin-5'-maleimide dye used in flow cytometric analysis for red cell membrane disorders.

Science.gov (United States)

Mehra, Simmi; Tyagi, Neetu; Dorwal, Pranav; Pande, Amit; Jain, Dharmendra; Sachdev, Ritesh; Raina, Vimarsh

2015-06-01

The eosin-5'-maleimide (EMA) binding test using flow cytometry is a common method to measure reduced mean channel fluorescence (MCF) of EMA-labeled red blood cells (RBCs) from patients with red cell membrane disorders. The basic principle of the EMA-RBC binding test involves the covalent binding of EMA to lysine-430 on the first extracellular loop of band 3 protein. In the present study, the MCF of EMA was analyzed for samples derived from 12 healthy volunteers (controls) to determine the stability (i.e., the percentage decrease in fluorescence) of EMA over a period of 1 year. Comparison of periodical MCF readings over time, that is, at 2-month intervals, showed that there were no significant changes in mean channel fluorescence for up to 6 months; however, there was a significant decrease in MCF at 8 months. For optimal dye utilization, EMA remained stable only for up to 6 months. Therefore, we recommend reconstitution of the dye every 6 months when implementing this test and storage at -80℃ in dark conditions.

Scaling-Laws of Flow Entropy with Topological Metrics of Water Distribution Networks

OpenAIRE

Giovanni Francesco Santonastaso; Armando Di Nardo; Michele Di Natale; Carlo Giudicianni; Roberto Greco

2018-01-01

Robustness of water distribution networks is related to their connectivity and topological structure, which also affect their reliability. Flow entropy, based on Shannon’s informational entropy, has been proposed as a measure of network redundancy and adopted as a proxy of reliability in optimal network design procedures. In this paper, the scaling properties of flow entropy of water distribution networks with their size and other topological metrics are studied. To such aim, flow entropy, ma...

Measurement of platelet aggregation, independently of patient platelet count

DEFF Research Database (Denmark)

Vinholt, P J; Frederiksen, H; Hvas, A-M

2017-01-01

with collagen-related peptide). Platelet aggregation had a negative predictive value of 100% for a bleeding tendency among patients. Conclusion The established platelet aggregation assay was applicable for thrombocytopenic patients, and improved the identification of bleeding risk.......Essentials •Platelet function may influence bleeding risk in thrombocytopenia, but useful tests are needed. •A flow cytometric platelet aggregation test independent of the patient platelet count was made. •Platelet aggregation was reduced in thrombocytopenic patients with hematological cancer....... •High platelet aggregation ruled out bleeding tendency in thrombocytopenic patients. Summary Background Methods for testing platelet aggregation in thrombocytopenia are lacking. Objective To establish a flow-cytometric test of in vitro platelet aggregation independently of the patient's platelet count...

Measurement of circulating cell-derived microparticles by flow cytometry: sources of variability within the assay.

Science.gov (United States)

Ayers, Lisa; Kohler, Malcolm; Harrison, Paul; Sargent, Ian; Dragovic, Rebecca; Schaap, Marianne; Nieuwland, Rienk; Brooks, Susan A; Ferry, Berne

2011-04-01

Circulating cell-derived microparticles (MPs) have been implicated in several disease processes and elevated levels are found in many pathological conditions. The detection and accurate measurement of MPs, although attracting widespread interest, is hampered by a lack of standardisation. The aim of this study was to establish a reliable flow cytometric assay to measure distinct subtypes of MPs in disease and to identify any significant causes of variability in MP quantification. Circulating MPs within plasma were identified by their phenotype (platelet, endothelial, leukocyte and annexin-V positivity (AnnV+). The influence of key variables (i.e. time between venepuncture and centrifugation, washing steps, the number of centrifugation steps, freezing/long-term storage and temperature of thawing) on MP measurement were investigated. Increasing time between venepuncture and centrifugation leads to increased MP levels. Washing samples results in decreased AnnV+MPs (P=0.002) and platelet-derived MPs (PMPs) (P=0.002). Double centrifugation of MPs prior to freezing decreases numbers of AnnV+MPs (P=0.0004) and PMPs (P=0.0004). A single freeze thaw cycle of samples led to an increase in AnnV+MPs (P=0.0020) and PMPs (P=0.0039). Long-term storage of MP samples at -80° resulted in decreased MP levels. This study found that minor protocol changes significantly affected MP levels. This is one of the first studies attempting to standardise a method for obtaining and measuring circulating MPs. Standardisation will be essential for successful development of MP technologies, allowing direct comparison of results between studies and leading to a greater understanding of MPs in disease. Crown Copyright © 2010. Published by Elsevier Ltd. All rights reserved.

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

African Journals Online (AJOL)

Jocky

2012-02-23

Feb 23, 2012 ... species. A systematic comparison of nuclear lysis buffers has been ... All experiments were carried out with 3 replicates (n = 3) per treatment. ... Na2EDTA, 0.5 mM spermine.4HCl, 80 mM KCl, 20 mM NaCl, 0.1%. (v/v) Triton ...

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)

A measurement device for electromagnetic flow tomography

Science.gov (United States)

Vauhkonen, M.; Hänninen, A.; Lehtikangas, O.

2018-01-01

Electromagnetic flow meters have succesfully been used in many industries to measure the mean flow velocity of conductive liquids. This technology works reliably in single phase flows with axisymmetric flow profiles but can be inaccurate with asymmetric flows, which are encountered, for example, in multiphase flows, pipe elbows and T-junctions. Some computational techniques and measurement devices with multiple excitation coils and measurement electrodes have recently been proposed to be used in cases of asymmetric flows. In earlier studies, we proposed a computational approach for electromagnetic flow tomography (EMFT) for estimating velocity fields utilizing several excitation coils and a set of measurement electrodes attached to the surface of the pipe. This approach has been shown to work well with simulated data but has not been tested extensively with real measurements. In this paper, an EMFT system with four excitation coils and 16 measurement electrodes is introduced. The system is capable of using both square wave and sinusoidal coil current excitations and all the coils can be excited individually, also enabling parallel excitations with multiple frequencies. The studies undertaken in the paper demonstrate that the proposed EMFT system, together with the earlier introduced velocity field reconstruction approach, is capable of producing reliable velocify field estimates in a laboratory environment with both axisymmetric and asymmetric single phase flows.

Early stage detection of chemotherapeutic effect on 203 GL glioma in mice as studied by P-31 NMR and flow cytometry

Energy Technology Data Exchange (ETDEWEB)

Itoh, Masamitsu; Yoshikawa, Koki; Nishikawa, Junichi; Iio, Masahiro; Shibui, Soichiro; Nomura, Kazuhiro; Saito, Hazime; Kodama, Masahiko

1988-08-01

The effect of chemotherapy against glioma in mouse was evaluated by /sup 31/P NMR spectroscopy and flow cytometry. We found that administration of ACNU or tegafur at a dose less than LD/sub 50/ resulted in the partial suppression of the ratio of inorganic phosphate (Pi)/phosphocreatine (PCr) and phosphomonoester (PME)/creatine phosphate (PCr) after 24 or 48 hr, although these ratios are usually increased together with growth of tumors. Flow cytometric analysis of glioma in vivo showed an accumulation in cells containing tetraploid DNA by G/sub 2/M block 24 - 48 hr after treatment. However, the change occurred at a period slightly later than that of the Pi/PCr ratio. In contrast, histological change was noted at eight days after administration. Hence, it is concluded that in vivo /sup 31/P NMR spectroscopy can detect a change in metabolic pathways in tumors as early as 24 - 48 hr after the administration of chemotherapeutic agents.

Reliable predictions of waste performance in a geologic repository

International Nuclear Information System (INIS)

Pigford, T.H.; Chambre, P.L.

1985-08-01

Establishing reliable estimates of long-term performance of a waste repository requires emphasis upon valid theories to predict performance. Predicting rates that radionuclides are released from waste packages cannot rest upon empirical extrapolations of laboratory leach data. Reliable predictions can be based on simple bounding theoretical models, such as solubility-limited bulk-flow, if the assumed parameters are reliably known or defensibly conservative. Wherever possible, performance analysis should proceed beyond simple bounding calculations to obtain more realistic - and usually more favorable - estimates of expected performance. Desire for greater realism must be balanced against increasing uncertainties in prediction and loss of reliability. Theoretical predictions of release rate based on mass-transfer analysis are bounding and the theory can be verified. Postulated repository analogues to simulate laboratory leach experiments introduce arbitrary and fictitious repository parameters and are shown not to agree with well-established theory. 34 refs., 3 figs., 2 tabs

Improving left ventricular segmentation in four-dimensional flow MRI using intramodality image registration for cardiac blood flow analysis.

Science.gov (United States)

Gupta, Vikas; Bustamante, Mariana; Fredriksson, Alexandru; Carlhäll, Carl-Johan; Ebbers, Tino

2018-01-01

Assessment of blood flow in the left ventricle using four-dimensional flow MRI requires accurate left ventricle segmentation that is often hampered by the low contrast between blood and the myocardium. The purpose of this work is to improve left-ventricular segmentation in four-dimensional flow MRI for reliable blood flow analysis. The left ventricle segmentations are first obtained using morphological cine-MRI with better in-plane resolution and contrast, and then aligned to four-dimensional flow MRI data. This alignment is, however, not trivial due to inter-slice misalignment errors caused by patient motion and respiratory drift during breath-hold based cine-MRI acquisition. A robust image registration based framework is proposed to mitigate such errors automatically. Data from 20 subjects, including healthy volunteers and patients, was used to evaluate its geometric accuracy and impact on blood flow analysis. High spatial correspondence was observed between manually and automatically aligned segmentations, and the improvements in alignment compared to uncorrected segmentations were significant (P  0.05). Our results demonstrate the efficacy of the proposed approach in improving left-ventricular segmentation in four-dimensional flow MRI, and its potential for reliable blood flow analysis. Magn Reson Med 79:554-560, 2018. © 2017 International Society for Magnetic Resonance in Medicine. © 2017 International Society for Magnetic Resonance in Medicine.

Analyzing the Cash Flow Profiles of Firms Using the Cash Flow Patterns Method

OpenAIRE

Aktaş, Rabia; Karğın, Sibel; Karğın, Mahmut

2012-01-01

Statement of cash flows that presents important information about firm’s power of cash generation and cash consumption is a reliable source of information for financial statement users. Therefore, financial reporting standards require cash flow statement to be reported and presented with the other financial statements such as statement of financial position, statement of comprehensive income, and statement of changes in equity. Turkish Accounting Standard 7 – “Statement of Cash Flows” (TMS 7 ...

A new method to measure flow in professional tasks – A FLOW-W questionnaire (FLOW at Work

Directory of Open Access Journals (Sweden)

Wolfigiel Beata

2017-06-01

Full Text Available The aim of the article is to present a new Polish tool for measuring the flow experience in professional tasks - a FLOW-W Questionnaire. The questionnaire was inspired by Csikszentmihalyi’s (1990 flow theory and flow in Bakker’s work (2008. On its basis a set of positions was established, on which subsequently an exploratory (study 1, N = 101 and confirmatory (study 2, N = 275 factor analysis was conducted. The analysis showed the possibility of a uni- or bifactorial solution. After checking the theoretical and empirical validity of both solutions, the unifactorial solution was adopted. The validity of the questionnaire was examined, i.a. by correlations with theoretically related variables: work engagement with the UWES questionnaire (Szabowska-Walaszczyk, Zawadzka, Wojtaś, 2011 and affect at work (Zalewska, 2002. The studies showed a significant positive correlation between flow at work and work engagement (0.84 and between flow and positive affect (0.74. The reliability of the questionnaire is very high; α = 0.96. The tool has very good psychometric properties.

Modeling, implementation, and validation of arterial travel time reliability : [summary].

Science.gov (United States)

2013-11-01

Travel time reliability (TTR) has been proposed as : a better measure of a facilitys performance than : a statistical measure like peak hour demand. TTR : is based on more information about average traffic : flows and longer time periods, thus inc...

Aortic blood flow subtraction: an alternative method for measuring total renal blood flow in conscious dogs

DEFF Research Database (Denmark)

Sandgaard, N C F; Andersen, J L; Holstein-Rathlou, N-H

2002-01-01

We have measured total renal blood flow (TRBF) as the difference between signals from ultrasound flow probes implanted around the aorta above and below the renal arteries. The repeatability of the method was investigated by repeated, continuous infusions of angiotensin II and endothelin-1 seven...... arterial blood pressure by 49% and decreased TRBF by 12%, providing an increase in renal vascular resistance of 69%. Dynamic analysis showed autoregulation of renal blood flow in the frequency range ... of TRBF by aortic blood flow subtraction is a practical and reliable method that allows direct comparison of excretory function and renal blood flow from two kidneys. The method also allows direct comparison between TRBF and flow in the caudal aorta....

Flow assurance studies for CO2 transport

NARCIS (Netherlands)

Veltin, J.; Belfroid, S.P.C.

2013-01-01

In order to compensate for the relative lack of experience of the CCTS community, Flow Assurance studies of new CO2 pipelines and networks are a very important step toward reliable operation. This report details a typical approach for Flow Assurance study of CO2 transport pipeline. Considerations to

Reliability engineering

International Nuclear Information System (INIS)

Lee, Chi Woo; Kim, Sun Jin; Lee, Seung Woo; Jeong, Sang Yeong

1993-08-01

This book start what is reliability? such as origin of reliability problems, definition of reliability and reliability and use of reliability. It also deals with probability and calculation of reliability, reliability function and failure rate, probability distribution of reliability, assumption of MTBF, process of probability distribution, down time, maintainability and availability, break down maintenance and preventive maintenance design of reliability, design of reliability for prediction and statistics, reliability test, reliability data and design and management of reliability.

Flow cytometric examination of apoptotic effect on brain tissue in postnatal period created by intrauterine oxcarbazepine and gabapentin exposure.

Science.gov (United States)

Erisgin, Z; Tekelioglu, Y

For epileptics, pregnancy contains the balance between no seizure period and antiepileptic use having the least teratogenicity risk. The purpose is to analyse with flow cytometry the apoptotic effects on postnatal brain tissue caused by prenatal use of second generation antiepileptics oxcarbazepine (OXC) and gabapentin (GBP) having different effect mechanisms. 30 (n = 5 each group) Wistar albino male rats (45-days-old) are used. First 3 groups are exposed to OXC (100 mg/kg/day), GBP (50 mg/kg/day), and saline, respectively on the 1st-5th prenatal days (preimplantation-implantation period) while the second 3 groups are exposed to the same substances on the 6th-15th prenatal days (organogenesis), respectively. After sacrifice, brain tissue samples were made into suspension with mechanic and enzymatic digestion and examined with flow cytometry. While apoptosis rate appeared high in rats exposed to OXC on the 1st-5th (p effect in three treatment groups, while difference was not significant for PSS and GBP groups (p = 0.847 and p = 0.934), apoptosis rate was significantly high for OXC on the 6th-15th days compared to the 1st-5th days (p < 0.001). It is observed that the use of OXC causes neurotoxicity during preimplantation, implantation and, especially, organogenesis period (neurogenesis) whereas GBP does not (Fig. 3, Ref. 32).

A network-flow based valve-switching aware binding algorithm for flow-based microfluidic biochips

DEFF Research Database (Denmark)

Tseng, Kai-Han; You, Sheng-Chi; Minhass, Wajid Hassan

2013-01-01

-flow based resource binding algorithm based on breadth-first search (BFS) and minimum cost maximum flow (MCMF) in architectural-level synthesis. The experimental results show that our methodology not only makes significant reduction of valve-switching activities but also diminishes the application completion......Designs of flow-based microfluidic biochips are receiving much attention recently because they replace conventional biological automation paradigm and are able to integrate different biochemical analysis functions on a chip. However, as the design complexity increases, a flow-based microfluidic...... biochip needs more chip-integrated micro-valves, i.e., the basic unit of fluid-handling functionality, to manipulate the fluid flow for biochemical applications. Moreover, frequent switching of micro-valves results in decreased reliability. To minimize the valve-switching activities, we develop a network...

Comparative analysis of minimal residual disease detection using four-color flow cytometry, consensus IgH-PCR, and quantitative IgH PCR in CLL after allogeneic and autologous stem cell transplantation.

Science.gov (United States)

Böttcher, S; Ritgen, M; Pott, C; Brüggemann, M; Raff, T; Stilgenbauer, S; Döhner, H; Dreger, P; Kneba, M

2004-10-01

The clinically most suitable method for minimal residual disease (MRD) detection in chronic lymphocytic leukemia is still controversial. We prospectively compared MRD assessment in 158 blood samples of 74 patients with CLL after stem cell transplantation (SCT) using four-color flow cytometry (MRD flow) in parallel with consensus IgH-PCR and ASO IgH real-time PCR (ASO IgH RQ-PCR). In 25 out of 106 samples (23.6%) with a polyclonal consensus IgH-PCR pattern, MRD flow still detected CLL cells, proving higher sensitivity of flow cytometry over PCR-genescanning with consensus IgH-primers. Of 92 samples, 14 (15.2%) analyzed in parallel by MRD flow and by ASO IgH RQ-PCR were negative by our flow cytometric assay but positive by PCR, thus demonstrating superior sensitivity of RQ-PCR with ASO primers. Quantitative MRD levels measured by both methods correlated well (r=0.93). MRD detection by flow and ASO IgH RQ-PCR were equally suitable to monitor MRD kinetics after allogeneic SCT, but the PCR method detected impending relapses after autologous SCT earlier. An analysis of factors that influence sensitivity and specificity of flow cytometry for MRD detection allowed to devise further improvements of this technique.

Flow boiling in expanding microchannels

CERN Document Server

Alam, Tamanna

2017-01-01

This Brief presents an up to date summary of details of the flow boiling heat transfer, pressure drop and instability characteristics; two phase flow patterns of expanding microchannels. Results obtained from the different expanding microscale geometries are presented for comparison and addition to that, comparison with literatures is also performed. Finally, parametric studies are performed and presented in the brief. The findings from this study could help in understanding the complex microscale flow boiling behavior and aid in the design and implementation of reliable compact heat sinks for practical applications.

Development of evaluation method on flow-induced vibration and corrosion of components in two-phase flow by coupled analysis. 1. Evaluation of effects of flow-induced vibration on structural material integrity

International Nuclear Information System (INIS)

Naitoh, Masanori; Uchida, Shunsuke; Koshizuka, Seiichi; Ninokata, Hisashi; Anahara, Naoki; Dosaki, Koji; Katono, Kenichi; Akiyama, Minoru; Saitoh, Hiroaki

2007-01-01

Problems in major components and structural materials in nuclear power plants have often been caused by flow induced vibration, corrosion and their overlapping effects. In order to establish safe and reliable plant operation, it is necessary to predict future problems for structural materials based on combined analyses of flow dynamics and corrosion and to mitigate them before they become serious issues for plant operation. An innovative method for flow induced vibration of structures in two phase flow by combined analyses of three dimensional flow dynamics and structures is to be introduced. (author)

Calculating Shocks In Flows At Chemical Equilibrium

Science.gov (United States)

Eberhardt, Scott; Palmer, Grant

1988-01-01

Boundary conditions prove critical. Conference paper describes algorithm for calculation of shocks in hypersonic flows of gases at chemical equilibrium. Although algorithm represents intermediate stage in development of reliable, accurate computer code for two-dimensional flow, research leading up to it contributes to understanding of what is needed to complete task.

PASSENGER FLOWS PREDICTION IN MAJOR TRANSPORTATION HUBS

Directory of Open Access Journals (Sweden)

O. O. Ozerova

2013-11-01

Full Text Available Purpose. An effective organization of passenger traffic, due to the reliability prediction of traffic flow in passenger transport hubs. In order to determine the parameters of prospective passenger transport areas it is essential to analyze the impact of various factors and determine the most influential ones. Methodology. The article presents the method of paired linear correlation for a more influential factors on passengers in intercity and commuter and possible use in predicting the linear regression equations. Passenger transport vessel areas and branches of industry are interconnected and are in the ratio of passengers and production. Findings. It is found that the coefficient of correlation is in complex dependence on the duration of the period of retrospective analysis. Evaluation of reliability correlation coefficients and coefficients of predictive models led to the conclusion that the population gives the most accurate prediction of passenger flows, providing account of changes in Ukraine during the period of transformation. Originality. Equations of dependence on the impact of macroeconomic indicators were obtained and the evaluation of the reliability results was received. Practical value. The results of analysis and calculations will make short-term forecasting of traffic flow.

Sodium component reliability data collection at CREDO

International Nuclear Information System (INIS)

Bott, T.F.; Haas, P.M.; Manning, J.J.

1979-01-01

The Centralized Reliability Data Organization (CREDO) has been established at Oak Ridge National Laboratory (ORNL) by the Department of Energy to provide a national center for collection, evaluation and dissemination of reliability data for advanced reactors. While the system is being developed and continuous data collection at the two U.S. reactor sites (EBR-II and FFTF) is being established, data on advanced reactor components which have been in use at U.S. test loops and experimental reactors have been collected and analyzed. Engineering, operating and event data on sodium valves, pumps, flow meters, rupture discs, heat exchangers and cold traps have been collected from more than a dozen sites. The results of analyses of the data performed to date are presented

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.

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.

Flow cytometric assessment of DNA damage in the fish Catla catla (Ham.) exposed to gamma radiation

International Nuclear Information System (INIS)

Anbumani, S.; Mohankumar, Mary N.; Selvanayagam, M.

2012-01-01

Environmental mutagens such as ionizing radiation and chemicals induce DNA damage in a wide variety of organisms. The International Commission on Radiological Protection (lCRP) has recently emphasized the need to protect non-human biota from the potential effects of ionizing radiation. Radiation exposures to non-humans can occur as a result of low-level radioactive discharges into the environment. Molecular genetic effects at low-level radiation exposures are largely unexplored and systematic studies using sensitive biomarkers are required to assess DNA damage in representative non-human species. The objective of the study was to detect DNA damage in the fish Catla catla exposed to gamma radiation using flow cytometry at different time intervals. Increases in the coefficient of variation (CV) of the G 0 /G 1 peak, indicating abnormal DNA distributions were observed in fish exposed to gamma radiation than in controls. Significant increase in the CV was observed from day 12-90 and thereafter decreased. This increase in CV might be due to DNA damage in the cell populations at G 0 /G 1 phase or deletions and duplications caused by improper repair of chromosomes in the cell-cycle machinery. Ionizing radiation induced cell-cycle perturbations and apoptosis were also observed after gamma radiation exposure. (author)

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.

Interlaboratory assessment of mitotic index by flow cytometry confirms superior reproducibility relative to microscopic scoring.

Science.gov (United States)

Roberts, D J; Spellman, R A; Sanok, K; Chen, H; Chan, M; Yurt, P; Thakur, A K; DeVito, G L; Murli, H; Stankowski, L F

2012-05-01

A flow cytometric procedure for determining mitotic index (MI) as part of the metaphase chromosome aberrations assay, developed and utilized routinely at Pfizer as part of their standard assay design, has been adopted successfully by Covance laboratories. This method, using antibodies against phosphorylated histone tails (H3PS10) and nucleic acid stain, has been evaluated by the two independent test sites and compared to manual scoring. Primary human lymphocytes were treated with cyclophosphamide, mitomycin C, benzo(a)pyrene, and etoposide at concentrations inducing dose-dependent cytotoxicity. Deming regression analysis indicates that the results generated via flow cytometry (FCM) were more consistent between sites than those generated via microscopy. Further analysis using the Bland-Altman modification of the Tukey mean difference method supports this finding, as the standard deviations (SDs) of differences in MI generated by FCM were less than half of those generated manually. Decreases in scoring variability owing to the objective nature of FCM, and the greater number of cells analyzed, make FCM a superior method for MI determination. In addition, the FCM method has proven to be transferable and easily integrated into standard genetic toxicology laboratory operations. Copyright © 2012 Wiley Periodicals, Inc.

Information flow a data bank preparation in nuclear power plant reliability information system

International Nuclear Information System (INIS)

Kolesa, K.; Vejvodova, I.

1983-01-01

In the year 1981 the reliability information system for nuclear power plants (ISS-JE) was established. The objective of the system is to make a statistical evaluation of the operation of nuclear power plants and to obtain information on the reliability of the equipment of nuclear power plants and the transmission of this information to manufacturers with the aim of inducing them to take corrective measures. The HP 1000 computer with the data base system IMAGE 100 is used which allows to process single queries and periodical outputs. The content of periodical outputs designed for various groups of subcontractors is briefly described and trends of the further development of the system indicated. (Ha)

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......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...

Distinct neutrophil subpopulations phenotype by flow cytometry in myelodysplastic syndromes.

Science.gov (United States)

Vikentiou, Myrofora; Psarra, Katerina; Kapsimali, Violetta; Liapis, Konstantinos; Michael, Michalis; Tsionos, Konstantinos; Lianidou, Evi; Papasteriades, Chryssa

2009-03-01

The cardinal feature of myelodysplastic syndromes (MDS) is dysplasia involving one or more myeloid cell lineages. In the present study, we used 4-color flow cytometric analysis to investigate dysgranulopoiesis in bone marrow specimens from 65 patients with MDS. The antigen expression patterns of total neutrophil granulocytes (TNG) and of the two distinct neutrophil granulocytic subpopulations (NGSs), NGS-1 (dimmer CD45 expression) and NGS-2 (stronger CD45 expression) identified on the side scatter (SS) vs. CD45-intensity plot, were studied. The neutrophil granulocytes from patients with MDS showed characteristic antigen expression aberrancies which were more pronounced in NGS-2 subpopulation. Studying separately the NGS-2 subpopulation with the CD16/MPO/LF combination, the low CD16(+)/MPO(+) and low CD16(+)/LF(+) percentages seemed to discriminate between lower-risk and higher-risk patients with MDS in most occasions. Furthermore, a detailed assessment of the NGS-1 and NGS-2 immunophenotypic patterns revealed early dysplastic changes, not otherwise observed by standard TNG analysis, especially in cases of lower-risk MDS.

On estimation of reliability of a nuclear power plant with tokamak reactor

International Nuclear Information System (INIS)

Klemin, A.I.; Smetannikov, V.P.; Shiverskij, E.A.

1982-01-01

The results of the analysis of INTOR plant reliability are presented. The first stage of the analysis consists in the calculation of the INTOR plant structural reliability factors (15 ibs main systems have been considered). For each system the failure flow parameter (W(1/h)) and operational readiness Ksub(r) have been determined which for the plant as a whole besides these factors-technological utilization coefficient Ksub(TU) and mean-cycles-between failures Tsub(o). The second stage of the reliability analysis consists in investigating methods of improving its reliability factors reratively to the one calculated at the first level stage. It is shown that the reliability of the whole plant to the most essential extent is determined by the power supply system reliability. The following as to the influence extent on the INTOR plant reliability is the cryogenic system. Calculations of the INTOR plant reliability factors have given the following values: W=4,5x10 -3 1/h. Tsub(o)=152 h, Ksub(r)=0,71, Ksub(TU)=o,4 g

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.

Quantitative analysis of gold and carbon nanoparticles in mammalian cells by flow cytometry light scattering

Energy Technology Data Exchange (ETDEWEB)

Zhou, Gang [Nanjing University, State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences (China); Liu, Naicheng; Wang, Zhenheng [Nanjing University, Department of Orthopedics, Jinling Hospital, School of Medicine (China); Shi, Tongguo; Gan, Jingjing; Wang, Zhenzhen; Zhang, Junfeng, E-mail: jfzhang@nju.edu.cn [Nanjing University, State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences (China)

2017-02-15

Nanoparticle-based applications for diagnostics and therapeutics have been extensively studied. These applications require a profound understanding of the fate of nanoparticles (NPs) in cellular environments. However, until now, few analytical methods are available and most of them rely on fluorescent properties or special elements of NPs; therefore, for NPs without observable optical properties or special elements, the existing methods are hardly applicable. In this study, we introduce a flow cytometry light scattering (FCLS)-based approach that quantifies in situ NPs accurately in mammalian cells. Continuous cells of heterogeneous human epithelial colorectal adenocarcinoma (Caco-2 cells), mouse peritoneal macrophages (MPM), and human adenocarcinomic alveolar basal epithelia (A549 cells) were cultured with NPs with certain concentrations and size. The intensity of the flow cytometric side scattered light, which indicates the quantity of NPs in the cells, was analyzed. The result shows an accurate size- and dose-dependent uptake of Au NPs (5, 30, 250 nm) in Caco-2 cells. The size- and dose- dependence of Au NPs (5, 30, 250 nm) and carbon NPs (50, 500 nm) in cells was validated by transmission electron microscope (TEM). This paper demonstrates the great potential of flow cytometry light scattering in the quantitative study of the size and dose effect on in situ metallic or non-metallic NPs in mammalian cells.

Quantitative analysis of gold and carbon nanoparticles in mammalian cells by flow cytometry light scattering

Science.gov (United States)

Zhou, Gang; Liu, Naicheng; Wang, Zhenheng; Shi, Tongguo; Gan, Jingjing; Wang, Zhenzhen; Zhang, Junfeng

2017-02-01

Nanoparticle-based applications for diagnostics and therapeutics have been extensively studied. These applications require a profound understanding of the fate of nanoparticles (NPs) in cellular environments. However, until now, few analytical methods are available and most of them rely on fluorescent properties or special elements of NPs; therefore, for NPs without observable optical properties or special elements, the existing methods are hardly applicable. In this study, we introduce a flow cytometry light scattering (FCLS)-based approach that quantifies in situ NPs accurately in mammalian cells. Continuous cells of heterogeneous human epithelial colorectal adenocarcinoma (Caco-2 cells), mouse peritoneal macrophages (MPM), and human adenocarcinomic alveolar basal epithelia (A549 cells) were cultured with NPs with certain concentrations and size. The intensity of the flow cytometric side scattered light, which indicates the quantity of NPs in the cells, was analyzed. The result shows an accurate size- and dose-dependent uptake of Au NPs (5, 30, 250 nm) in Caco-2 cells. The size- and dose- dependence of Au NPs (5, 30, 250 nm) and carbon NPs (50, 500 nm) in cells was validated by transmission electron microscope (TEM). This paper demonstrates the great potential of flow cytometry light scattering in the quantitative study of the size and dose effect on in situ metallic or non-metallic NPs in mammalian cells.

An improved flow cytometric method using FACS lysing solution for measurement of ZAP-70 expression in B-cell chronic lymphocytic leukemia

NARCIS (Netherlands)

Bekkema, Roelof; Tadema, Afke; Daenen, Simon M. G. J.; Kluin-Nelemans, Hanneke C.; Mulder, Andre B.

Background: B-cell expression of ZAP-70, normally expressed in T and NK cells, correlates with poor prognosis in B-CLL. Poor discrimination between ZAP-70 positive and negative cells hampers routine application of flow cytometry. We examined the usefulness of FACS Lysing Solution. Methods: ZAP-70

Reducing variability of workforce as a tool to improve plan reliability

DEFF Research Database (Denmark)

Wandahl, Søren; Yicheng, S.; Zygmunt, K. J.

Variability of flow is recognized as the greatest obstacle to production management. Since the work flow and labour flow are two dominators of work performance, it is important to manage them simultaneously. The objective of this paper is to examine whether by reducing the variance of a labour flow......, a plan reliability can be improved, therefore, three different construction labour data sets have been examined by utilizing Monte Carlo Simulation, to analyze the probability to finish simulated projects within a certain time. The research findings revealed that reducing variance in the workforce flow...... does not necessarily shorten the project length, nevertheless it increases probability to finish the tasks within a critical path duration. Additionally, it was concluded, that reducing the variance of crew allocation can improve the productivity....

Reducing Variability of Workforce as a Tool to Improve Plan Reliability

DEFF Research Database (Denmark)

Shen, Yicheng; Zygmunt, Katarzyna Julia; Wandahl, Søren

2017-01-01

Variability of flow is recognized as one of the greatest obstacles to production management. Since the work flow and labour flow are two dominators of work performance, it is important to manage them simultaneously. The objective of this paper is to examine if an increased plan reliability could...... of the workforce flow does not necessarily shorten the project length, nevertheless it increases probability to finish the tasks within a critical path duration. Additionally, it was concluded, that reducing the variance of crew allocation can improve the productivity....... be reached by reducing the variance of a labour flow. Therefore, three different construction labour data sets have been examined by utilizing Monte Carlo Simulation, to analyze the probability to finish simulated projects within a certain time. The research findings revealed that reducing variance...

Methodology for local verification of flow regimes in fuel assemblies charts

International Nuclear Information System (INIS)

Igor, Sharaevsky; Elena, Sharaevskaya; Domashev, E.D.; Alexander, Arkhypov; Vladimir, Kolochko

2003-01-01

The best estimate thermal hydraulic codes describe adequately two-phase flows in nuclear energy facilities if there is proper system of closed relations. It could be obtained from the reliable information on structure forms of two-phase flows, its boundaries and reliable regime charts. In the paper the methodology of automatic recognition of the boundaries of the main types of two phase flows for rod fuel assemblies is presented. The methodology is based on definition of thermal hydraulic parameters distribution in experimental fuel assembly. The measurements were carried out using ASD signals of acoustic noise. In the paper data on two-phase flow regimes boundaries recognition especially low boundaries of bubble flow are summarized for experimental fuel assembly. The methodology of flow regimes charts applied to recognition of upper boundaries of boiling crisis regime was verificated. The satisfactory coincidence with experimental results have been shown. (author)

Laboratory Tests of Multiplex Detection of PCR Amplicons Using the Luminex 100 Flow Analyzer

Energy Technology Data Exchange (ETDEWEB)

Venkateswaran, K.S.; Nasarabadi, S.; Langlois, R.G.

2000-05-05

Lawrence Livermore National Laboratory (LLNL) demonstrated the power of flow cytometry in detecting the biological agents simulants at JFT III. LLNL pioneered in the development of advanced nucleic acid analyzer (ANM) for portable real time identification. Recent advances in flow cytometry provide a means for multiplexed nucleic acid detection and immunoassay of pathogenic microorganisms. We are presently developing multiplexed immunoassays for the simultaneous detection of different simulants. Our goal is to build an integrated instrument for both nucleic acid analysis and immuno detection. In this study we evaluated the Luminex LX 100 for concurrent identification of more than one PCR amplified product. ANAA has real-time Taqman fluorescent detection capability for rapid identification of field samples. However, its multiplexing ability is limited by the combination of available fluorescent labels. Hence integration of ANAA with flow cytometry can give the rapidity of ANAA amplification and the multiplex capability of flow cytometry. Multiplexed flow cytometric analysis is made possible using a set of fluorescent latex microsphere that are individually identified by their red and infrared fluorescence. A green fluorochrome is used as the assay signal. Methods were developed for the identification of specific nucleic acid sequences from Bacillus globigii (Bg), Bacillus thuringensis (Bt) and Erwinia herbicola (Eh). Detection sensitivity using different reporter fluorochromes was tested with the LX 100, and also different assay formats were evaluated for their suitability for rapid testing. A blind laboratory trial was carried out December 22-27, 1999 to evaluate bead assays for multiplex identification of Bg and Bt PCR products. This report summarizes the assay development, fluorochrome comparisons, and the results of the blind trial conducted at LLNL for the laboratory evaluation of the LX 100 flow analyzer.

Detection of mycoplasmas in goat milk by flow cytometry.

Science.gov (United States)

Assunção, Patricia; Davey, Hazel M; Rosales, Ruben S; Antunes, Nuno T; de la Fe, Christian; Ramirez, Ana S; de Galarreta, Carlos M Ruiz; Poveda, Jose B

2007-12-01

The detection of mycoplasma in milk can be performed by either culture techniques or polymerase chain reaction (PCR) based methods. Although PCR can reduce the average diagnostic time to 5 h in comparison with the several days for the isolation of the agent, there is still a need to develop methods, which could give earlier results. For this purpose, we tested the ability of flow cytometry (FC) to detect mycoplasmas in milk samples. Milk samples inoculated with four different mycoplasmas, Mycoplasma agalactiae, Mycoplasma putrefaciens, Mycoplasma capricolum subsp. Capricolum, or Mycoplasma mycoides subsp. mycoides large-colony type, known to cause contagious agalactia in goats, were stained with the DNA stain SYBR Green I and analyzed by FC. Three goat milk samples, from which mycoplasmas have been isolated in broth medium were also analyzed. All mycoplasmas were easily distinguished from debris of milk samples, but it was not possible to distinguish between the different mycoplasma species. In our conditions, the detection limit of the technique was of the order of 10(3)-10(4) cells ml(-1). Furthermore, mycoplasmas were also distinguished from Staphylococcus aureus. FC together with SYBR Green I was able to distinguish between mycoplasma cells and debris present in milk samples and gave results in 20-30 min. This is an important first step in developing a robust, routine flow cytometric method for the detection of mycoplasmas in milk samples. (c) 2007 International Society for Analytical Cytology

Systems reliability/structural reliability

International Nuclear Information System (INIS)

Green, A.E.

1980-01-01

The question of reliability technology using quantified techniques is considered for systems and structures. Systems reliability analysis has progressed to a viable and proven methodology whereas this has yet to be fully achieved for large scale structures. Structural loading variants over the half-time of the plant are considered to be more difficult to analyse than for systems, even though a relatively crude model may be a necessary starting point. Various reliability characteristics and environmental conditions are considered which enter this problem. The rare event situation is briefly mentioned together with aspects of proof testing and normal and upset loading conditions. (orig.)

Financial reporting reliability concept and impact factors in its determination

Directory of Open Access Journals (Sweden)

Vygivska I.М.

2017-03-01

Full Text Available The article sets the goal to assess the reliability of financial reporting essence and summarize the factors that affect it. The article grounds the reliability of financial reporting urgency and characterizes its dual nature (on one hand, it is accounting containing reliable, reasonable data, characterized by the absence of errors and can be correctly perceived by users, on the other hand, reliable financial statements are considered to be formed according to the rules regulated by normative documents. It is proved that making decisions by interested users based on financial statements is closely related to information risk that arises because of the restrictions affect the flow of information on which management decisions are taken (accounting information that is difficult to understand by unskilled users, the impact of human factor, the level of information importance. The author establishes that the reliability of financial reporting is affected by many factors whose influence can be seen both outside, and within an enterprise. The main factors include: informational, political, legal, human, organizational and financial and they should be taken into account while making management decisions.

CERTS: Consortium for Electric Reliability Technology Solutions - Research Highlights

Energy Technology Data Exchange (ETDEWEB)

Eto, Joseph

2003-07-30

Historically, the U.S. electric power industry was vertically integrated, and utilities were responsible for system planning, operations, and reliability management. As the nation moves to a competitive market structure, these functions have been disaggregated, and no single entity is responsible for reliability management. As a result, new tools, technologies, systems, and management processes are needed to manage the reliability of the electricity grid. However, a number of simultaneous trends prevent electricity market participants from pursuing development of these reliability tools: utilities are preoccupied with restructuring their businesses, research funding has declined, and the formation of Independent System Operators (ISOs) and Regional Transmission Organizations (RTOs) to operate the grid means that control of transmission assets is separate from ownership of these assets; at the same time, business uncertainty, and changing regulatory policies have created a climate in which needed investment for transmission infrastructure and tools for reliability management has dried up. To address the resulting emerging gaps in reliability R&D, CERTS has undertaken much-needed public interest research on reliability technologies for the electricity grid. CERTS' vision is to: (1) Transform the electricity grid into an intelligent network that can sense and respond automatically to changing flows of power and emerging problems; (2) Enhance reliability management through market mechanisms, including transparency of real-time information on the status of the grid; (3) Empower customers to manage their energy use and reliability needs in response to real-time market price signals; and (4) Seamlessly integrate distributed technologies--including those for generation, storage, controls, and communications--to support the reliability needs of both the grid and individual customers.

Reliability Engineering

International Nuclear Information System (INIS)

Lee, Sang Yong

1992-07-01

This book is about reliability engineering, which describes definition and importance of reliability, development of reliability engineering, failure rate and failure probability density function about types of it, CFR and index distribution, IFR and normal distribution and Weibull distribution, maintainability and movability, reliability test and reliability assumption in index distribution type, normal distribution type and Weibull distribution type, reliability sampling test, reliability of system, design of reliability and functionality failure analysis by FTA.

Optimised Renormalisation Group Flows

CERN Document Server

Litim, Daniel F

2001-01-01

Exact renormalisation group (ERG) flows interpolate between a microscopic or classical theory and the corresponding macroscopic or quantum effective theory. For most problems of physical interest, the efficiency of the ERG is constrained due to unavoidable approximations. Approximate solutions of ERG flows depend spuriously on the regularisation scheme which is determined by a regulator function. This is similar to the spurious dependence on the ultraviolet regularisation known from perturbative QCD. Providing a good control over approximated ERG flows is at the root for reliable physical predictions. We explain why the convergence of approximate solutions towards the physical theory is optimised by appropriate choices of the regulator. We study specific optimised regulators for bosonic and fermionic fields and compare the optimised ERG flows with generic ones. This is done up to second order in the derivative expansion at both vanishing and non-vanishing temperature. An optimised flow for a ``proper-time ren...

Study of flow behavior in all-vanadium redox flow battery using spatially resolved voltage distribution

Science.gov (United States)

Bhattarai, Arjun; Wai, Nyunt; Schweiss, Rüdiger; Whitehead, Adam; Scherer, Günther G.; Ghimire, Purna C.; Nguyen, Tam D.; Hng, Huey Hoon

2017-08-01

Uniform flow distribution through the porous electrodes in a flow battery cell is very important for reducing Ohmic and mass transport polarization. A segmented cell approach can be used to obtain in-situ information on flow behaviour, through the local voltage or current mapping. Lateral flow of current within the thick felts in the flow battery can hamper the interpretation of the data. In this study, a new method of segmenting a conventional flow cell is introduced, which for the first time, splits up both the porous felt as well as the current collector. This dual segmentation results in higher resolution and distinct separation of voltages between flow inlet to outlet. To study the flow behavior for an undivided felt, monitoring the OCV is found to be a reliable method, instead of voltage or current mapping during charging and discharging. Our approach to segmentation is simple and applicable to any size of the cell.

Role of recent research in improving check valve reliability at nuclear power plants

International Nuclear Information System (INIS)

Kalsi, M.S.; Horst, C.L.; Wang, J.K.; Sharma, V.

1990-01-01

Check valve failures at nuclear power plants in recent years have led to serious safety concerns, and caused extensive damage to other plant components which had a significant impact on plant availability. In order to understand the failure mechanism and improve the reliability of check valves, a systematic research effort was proposed by Kalsi Engineering, Inc. to U.S. Nuclear Regulatory Commission (NRC). The overall goal of the research was to develop models for predicting the performance and degradation of swing check valves in nuclear power plant systems so that appropriate preventive maintenance or design modifications can be performed to improve the reliability of check valves. Under Phase I of this research, a large matrix of tests was run with instrumented swing check valves to determine the stability of the disc under a variety of upstream flow disturbances, covering a wide range of disc stop positions and flow velocities in two different valve sizes. The goals of Phase II research were to develop predictive models which quantify the anticipated degradation of swing check valves that have flow disturbances closely upstream of the valve and are operating under flow velocities that do not result in full disc opening. This research allows the inspection/maintenance activities to be focussed on those check valves that are more likely to suffer premature degradation. The quantitative wear and fatigue prediction methodology can be used to develop a sound preventive maintenance program. The results of the research also show the improvements in check valve performance/reliability that can be achieved by certain modifications in the valve design

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

Reliability

OpenAIRE

Condon, David; Revelle, William

2017-01-01

Separating the signal in a test from the irrelevant noise is a challenge for all measurement. Low test reliability limits test validity, attenuates important relationships, and can lead to regression artifacts. Multiple approaches to the assessment and improvement of reliability are discussed. The advantages and disadvantages of several different approaches to reliability are considered. Practical advice on how to assess reliability using open source software is provided.

Evaluation of Network Reliability for Computer Networks with Multiple Sources

Directory of Open Access Journals (Sweden)

Yi-Kuei Lin

2012-01-01

Full Text Available Evaluating the reliability of a network with multiple sources to multiple sinks is a critical issue from the perspective of quality management. Due to the unrealistic definition of paths of network models in previous literature, existing models are not appropriate for real-world computer networks such as the Taiwan Advanced Research and Education Network (TWAREN. This paper proposes a modified stochastic-flow network model to evaluate the network reliability of a practical computer network with multiple sources where data is transmitted through several light paths (LPs. Network reliability is defined as being the probability of delivering a specified amount of data from the sources to the sink. It is taken as a performance index to measure the service level of TWAREN. This paper studies the network reliability of the international portion of TWAREN from two sources (Taipei and Hsinchu to one sink (New York that goes through a submarine and land surface cable between Taiwan and the United States.

Flow cytometry-assisted rapid isolation of recombinant Plasmodium berghei parasites exemplified by functional analysis of aquaglyceroporin

Science.gov (United States)

Kenthirapalan, Sanketha; Waters, Andrew P.; Matuschewski, Kai; Kooij, Taco W.A.

2012-01-01

The most critical bottleneck in the generation of recombinant Plasmodium berghei parasites is the mandatory in vivo cloning step following successful genetic manipulation. This study describes a new technique for rapid selection of recombinant P. berghei parasites. The method is based on flow cytometry to isolate isogenic parasite lines and represents a major advance for the field, in that it will speed the generation of recombinant parasites as well as cut down on animal use significantly. High expression of GFP during blood infection, a prerequisite for robust separation of transgenic lines by flow cytometry, was achieved. Isogenic recombinant parasite populations were isolated even in the presence of a 100-fold excess of wild-type (WT) parasites. Aquaglyceroporin (AQP) loss-of-function mutants and parasites expressing a tagged AQP were generated to validate this approach. aqp− parasites grow normally within the WT phenotypic range during blood infection of NMRI mice. Similarly, colonization of the insect vector and establishment of an infection after mosquito transmission were unaffected, indicating that AQP is dispensable for life cycle progression in vivo under physiological conditions, refuting its use as a suitable drug target. Tagged AQP localized to perinuclear structures and not the parasite plasma membrane. We suggest that flow-cytometric isolation of isogenic parasites overcomes the major roadblock towards a genome-scale repository of mutant and transgenic malaria parasite lines. PMID:23137753

MSET modeling of Crystal River-3 venturi flow meters

International Nuclear Information System (INIS)

Bockhorst, F. K.; Gross, K. C.; Herzog, J. P.; Wegerich, S. W.

1998-01-01

The analysis of archived Crystal River-3 feedwater flow data reveals a slow and steady degradation of the flow meter measurements during the 1992/1993 operating cycle. MSET can reliably estimate the true flow rate and quantify the degree of departure between the indicated signal and the true flow rate with high accuracy. The MSET computed flow rate could, in principle, be used to provide an improved estimate of the reactor power and hence avoid the revenue loss associated with derating the reactor based on a faulty feedwater flow rate indication

Reactor coolant flow measurements at Point Lepreau

International Nuclear Information System (INIS)

Brenciaglia, G.; Gurevich, Y.; Liu, G.

1996-01-01

The CROSSFLOW ultrasonic flow measurement system manufactured by AMAG is fully proven as reliable and accurate when applied to large piping in defined geometries for such applications as feedwater flows measurement. Its application to direct reactor coolant flow (RCF) measurements - both individual channel flows and bulk flows such as pump suction flow - has been well established through recent work by AMAG at Point Lepreau, with application to other reactor types (eg. PWR) imminent. At Point Lepreau, Measurements have been demonstrated at full power; improvements to consistently meet ±1% accuracy are in progress. The development and recent customization of CROSSFLOW to RCF measurement at Point Lepreau are described in this paper; typical measurement results are included. (author)

Composite system reliability evaluation by stochastic calculation of system operation

Energy Technology Data Exchange (ETDEWEB)

Haubrick, H -J; Hinz, H -J; Landeck, E [Dept. of Power Systems and Power Economics (Germany)

1994-12-31

This report describes a new developed probabilistic approach for steady-state composite system reliability evaluation and its exemplary application to a bulk power test system. The new computer program called PHOENIX takes into consideration transmission limitations, outages of lines and power stations and, as a central element, a highly sophisticated model to the dispatcher performing remedial actions after disturbances. The kernel of the new method is a procedure for optimal power flow calculation that has been specially adapted for the use in reliability evaluations under the above mentioned conditions. (author) 11 refs., 8 figs., 1 tab.

RTE - Reliability report 2016

International Nuclear Information System (INIS)

2017-06-01

Every year, RTE produces a reliability report for the past year. This document lays out the main factors that affected the electrical power system's operational reliability in 2016 and the initiatives currently under way intended to ensure its reliability in the future. Within a context of the energy transition, changes to the European interconnected network mean that RTE has to adapt on an on-going basis. These changes include the increase in the share of renewables injecting an intermittent power supply into networks, resulting in a need for flexibility, and a diversification in the numbers of stakeholders operating in the energy sector and changes in the ways in which they behave. These changes are dramatically changing the structure of the power system of tomorrow and the way in which it will operate - particularly the way in which voltage and frequency are controlled, as well as the distribution of flows, the power system's stability, the level of reserves needed to ensure supply-demand balance, network studies, assets' operating and control rules, the tools used and the expertise of operators. The results obtained in 2016 are evidence of a globally satisfactory level of reliability for RTE's operations in somewhat demanding circumstances: more complex supply-demand balance management, cross-border schedules at interconnections indicating operation that is closer to its limits and - most noteworthy - having to manage a cold spell just as several nuclear power plants had been shut down. In a drive to keep pace with the changes expected to occur in these circumstances, RTE implemented numerous initiatives to ensure high levels of reliability: - maintaining investment levels of euro 1.5 billion per year; - increasing cross-zonal capacity at borders with our neighbouring countries, thus bolstering the security of our electricity supply; - implementing new mechanisms (demand response, capacity mechanism, interruptibility, etc.); - involvement in tests or projects

Diacerein retards cell growth of chondrosarcoma cells at the G2/M cell cycle checkpoint via cyclin B1/CDK1 and CDK2 downregulation

International Nuclear Information System (INIS)

Lohberger, Birgit; Leithner, Andreas; Stuendl, Nicole; Kaltenegger, Heike; Kullich, Werner; Steinecker-Frohnwieser, Bibiane

2015-01-01

Chondrosarcoma is characterized for its lack of response to conventional cytotoxic chemotherapy, propensity for developing lung metastases, and low rates of survival. Research within the field of development and expansion of new treatment options for unresectable or metastatic diseases is of particular priority. Diacerein, a symptomatic slow acting drug in osteoarthritis (SYSADOA), implicates a therapeutic benefit for the treatment of chondrosarcoma by an antitumor activity. After treatment with diacerein the growth behaviour of the cells was analyzed with the xCELLigence system and MTS assay. Cell cycle was examined using flow cytometric analysis, RT-PCR, and western blot analysis of specific checkpoint regulators. The status for phosophorylation of mitogen-activated protein kinases (MAPKs) was analyzed with a proteome profiler assay. In addition, the possible impact of diacerein on apoptosis was investigated using cleaved caspase 3 and Annexin V/PI flow cytometric analysis. Diacerein decreased the cell viability and the cell proliferation in two different chondrosarcoma cell lines in a dose dependent manner. Flow cytometric analysis showed a classical G2/M arrest. mRNA and protein analysis revealed that diacerein induced a down-regulation of the cyclin B1-CDK1 complex and a reduction in CDK2 expression. Furthermore, diacerein treatment increased the phosphorylation of p38α and p38β MAPKs, and Akt1, Akt2, and Akt 3 in SW-1353, whereas in Cal-78 the opposite effect has been demonstrated. These observations accordingly to our cell cycle flow cytometric analysis and protein expression data may explain the G2/M phase arrest. In addition, no apoptotic induction after diacerein treatment, neither in the Cal-78 nor in the SW-1353 cell line was observed. Our results demonstrate for the first time that the SYSADOA diacerein decreased the viability of human chondrosarcoma cells and induces G2/M cell cycle arrest by CDK1/cyclin B1 down-regulation

Stochastic network interdiction optimization via capacitated network reliability modeling and probabilistic solution discovery

International Nuclear Information System (INIS)

Ramirez-Marquez, Jose Emmanuel; Rocco S, Claudio M.

2009-01-01

This paper introduces an evolutionary optimization approach that can be readily applied to solve stochastic network interdiction problems (SNIP). The network interdiction problem solved considers the minimization of the cost associated with an interdiction strategy such that the maximum flow that can be transmitted between a source node and a sink node for a fixed network design is greater than or equal to a given reliability requirement. Furthermore, the model assumes that the nominal capacity of each network link and the cost associated with their interdiction can change from link to link and that such interdiction has a probability of being successful. This version of the SNIP is for the first time modeled as a capacitated network reliability problem allowing for the implementation of computation and solution techniques previously unavailable. The solution process is based on an evolutionary algorithm that implements: (1) Monte-Carlo simulation, to generate potential network interdiction strategies, (2) capacitated network reliability techniques to analyze strategies' source-sink flow reliability and, (3) an evolutionary optimization technique to define, in probabilistic terms, how likely a link is to appear in the final interdiction strategy. Examples for different sizes of networks are used throughout the paper to illustrate the approach

Flow Cytometry Enables Multiplexed Measurements of Genetically Encoded Intramolecular FRET Sensors Suitable for Screening.

Science.gov (United States)

Doucette, Jaimee; Zhao, Ziyan; Geyer, Rory J; Barra, Melanie M; Balunas, Marcy J; Zweifach, Adam

2016-07-01

Genetically encoded sensors based on intramolecular FRET between CFP and YFP are used extensively in cell biology research. Flow cytometry has been shown to offer a means to measure CFP-YFP FRET; we suspected it would provide a unique way to conduct multiplexed measurements from cells expressing different FRET sensors, which is difficult to do with microscopy, and that this could be used for screening. We confirmed that flow cytometry accurately measures FRET signals using cells transiently transfected with an ERK activity reporter, comparing responses measured with imaging and cytometry. We created polyclonal long-term transfectant lines, each expressing a different intramolecular FRET sensor, and devised a way to bar-code four distinct populations of cells. We demonstrated the feasibility of multiplexed measurements and determined that robust multiplexed measurements can be conducted in plate format. To validate the suitability of the method for screening, we measured responses from a plate of bacterial extracts that in unrelated experiments we had determined contained the protein kinase C (PKC)-activating compound teleocidin A-1. The multiplexed assay correctly identifying the teleocidin A-1-containing well. We propose that multiplexed cytometric FRET measurements will be useful for analyzing cellular function and for screening compound collections. © 2016 Society for Laboratory Automation and Screening.

Panel development for multicolor flow-cytometry testing of proliferation and immunophenotype in hMSCs.

Science.gov (United States)

Bradford, Jolene A; Clarke, Scott T

2011-01-01

Adult human mesenchymal stem cells (hMSC) are rare fibroblast-like cells capable of differentiation into a variety of cell tissues which include bone, cartilage, muscle, ligament, tendon, and adipose. Normal adult bone marrow and adipose tissue are the most common sources of these cells. The International Society for Cellular Therapy (ISCT) has proposed a set of standards to define hMSC for laboratory investigations and preclinical studies: adherence to plastic in standard culture conditions; in vitro differentiation into osteoblasts, adipocytes, and chondroblasts; and specific surface antigen expression. Direct measurement of proliferation combined with simultaneous detection of the ISCT-consensus immunophenotypic profile provides data that is used to determine the differentiation status and health of the cells. Flow cytometry provides a powerful technology that is routinely used to simultaneously and rapidly measure multiple parameters in a single sample. This chapter describes a flow cytometric panel for the simultaneous detection of immunophenotypic profile, proliferative capacity, and DNA content measurement in hMSC. Because a relatively small number of cells are needed with this approach, measurements can be made with minimal impact on expansion potential. The ability to assess antigen expression and proliferative status enables the investigator to make informed decisions on expansion and harvesting.

Suppression of panel flutter of near-space aircraft based on non-probabilistic reliability theory

Directory of Open Access Journals (Sweden)

Ye-Wei Zhang

2016-03-01

Full Text Available The vibration active control of the composite panels with the uncertain parameters in the hypersonic flow is studied using the non-probabilistic reliability theory. Using the piezoelectric patches as active control actuators, dynamic equations of panel are established by finite element method and Hamilton’s principle. And the control model of panel with uncertain parameters is obtained. According to the non-probabilistic reliability index, and besides being based on H∞ robust control theory and non-probabilistic reliability theory, the non-probabilistic reliability performance function is given. Moreover, the relationships between the robust controller and H∞ performance index and reliability are established. Numerical results show that the control method under the influence of reliability, H∞ performance index, and approaching velocity is effective to the vibration suppression of panel in the whole interval of uncertain parameters.

Social network analysis via multi-state reliability and conditional influence models

International Nuclear Information System (INIS)

Schneider, Kellie; Rainwater, Chase; Pohl, Ed; Hernandez, Ivan; Ramirez-Marquez, Jose Emmanuel

2013-01-01

This paper incorporates multi-state reliability measures into the assessment of a social network in which influence is treated as a multi-state commodity that flows through the network. The reliability of the network is defined as the probability that at least a certain level of influence reaches an intended target. We consider an individual's influence level as a function of the influence levels received from preceding actors in the network. We define several communication functions which describe the level of influence a particular actor will pass along to other actors within the network. Illustrative examples are presented, and the network reliability under the various communication influence levels is computed using exhaustive enumeration for a small example and Monte Carlo simulation for larger, more realistic sized examples.

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)

A method to analyze, sort, and retain viability of obligate anaerobic microorganisms from complex microbial communities.

Science.gov (United States)

Thompson, Anne W; Crow, Matthew J; Wadey, Brian; Arens, Christina; Turkarslan, Serdar; Stolyar, Sergey; Elliott, Nicholas; Petersen, Timothy W; van den Engh, Ger; Stahl, David A; Baliga, Nitin S

2015-10-01

A high speed flow cytometric cell sorter was modified to maintain a controlled anaerobic environment. This technology enabled coupling of the precise high-throughput analytical and cell separation capabilities of flow cytometry to the assessment of cell viability of evolved lineages of obligate anaerobic organisms from cocultures. Copyright © 2015. Published by Elsevier B.V.

A tripartite mode of action approach for investigating the impact of aneugens on tubulin polymerization.

Science.gov (United States)

Stock, Valerie; Sutter, Andreas; Raschke, Marian; Queisser, Nina

2018-04-01

Chemical-induced disruption of the cellular microtubule network is one key mechanism of aneugenicity. Since recent data indicate that genotoxic effects of aneugens show nonlinear dose-response relationships, margins of safety can be derived with the ultimate goal to perform a risk assessment for the support of drug development. Furthermore, microtubule-interacting compounds are widely used for cancer treatment. While there is a need to support the risk assessment of tubulin-interacting chemicals using reliable mechanistic assays, no standard assays exist to date in regulatory genotoxicity testing for the distinction of aneugenic mechanisms. Recently reported methods exclusively rely on either biochemical, morphological, or cytometric endpoints. Since data requirements for the diverse fields of application of those assays differ strongly, the use of multiple assays for a correct classification of aneugens is ideal. We here report a tripartite mode of action approach comprising a cell-free biochemical polymerization assay and the cell-based methods cellular imaging and flow cytometry. The biochemical assay measures tubulin polymerization over time whereas the two cell-based assays quantify tubulin polymer mass. We herein show that the flow cytometric method yielded IC 50 values for tubulin destabilizers and EC 50 values for tubulin stabilizers as well as cell cycle information. In contrast, cellular imaging complemented these findings with characteristic morphological patterns. Biochemical analysis yielded kinetic information on tubulin polymerization. This multiplex approach is able to create holistic effect profiles which can be individually customized to the research question with regard to quality, quantity, usability, and economy. Environ. Mol. Mutagen. 59:188-201, 2018. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

System Reliability Evaluation of Data Transmission in Commercial Banks with Multiple Branches

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Yi-Kuei Lin

2014-01-01

Full Text Available The main purpose of this paper is to assess the system reliability of electronic transaction data transmissions made by commercial banks in terms of stochastic flow network. System reliability is defined as the probability of demand satisfaction and it can be used to measure quality of service. In this paper, we study the system reliability of data transmission from the headquarters of a commercial bank to its multiple branches. The network structure of the bank and the probability of successful data transmission are obtained through the collection of real data. The system reliability, calculated using the minimal path method and the recursive sum of disjoint products algorithm, provides banking managers with a view to comprehend the current state of the entire system. Besides, the system reliability can be used not only as a measurement of quality of service, but also an improvement reference of the system by adopting sensitivity analysis.

Accounting for Model Uncertainties Using Reliability Methods - Application to Carbon Dioxide Geologic Sequestration System. Final Report

International Nuclear Information System (INIS)

Mok, Chin Man; Doughty, Christine; Zhang, Keni; Pruess, Karsten; Kiureghian, Armen; Zhang, Miao; Kaback, Dawn

2010-01-01

A new computer code, CALRELTOUGH, which uses reliability methods to incorporate parameter sensitivity and uncertainty analysis into subsurface flow and transport models, was developed by Geomatrix Consultants, Inc. in collaboration with Lawrence Berkeley National Laboratory and University of California at Berkeley. The CALREL reliability code was developed at the University of California at Berkely for geotechnical applications and the TOUGH family of codes was developed at Lawrence Berkeley National Laboratory for subsurface flow and tranport applications. The integration of the two codes provides provides a new approach to deal with uncertainties in flow and transport modeling of the subsurface, such as those uncertainties associated with hydrogeology parameters, boundary conditions, and initial conditions of subsurface flow and transport using data from site characterization and monitoring for conditioning. The new code enables computation of the reliability of a system and the components that make up the system, instead of calculating the complete probability distributions of model predictions at all locations at all times. The new CALRELTOUGH code has tremendous potential to advance subsurface understanding for a variety of applications including subsurface energy storage, nuclear waste disposal, carbon sequestration, extraction of natural resources, and environmental remediation. The new code was tested on a carbon sequestration problem as part of the Phase I project. Phase iI was not awarded.

A reliable sewage quality abnormal event monitoring system.

Science.gov (United States)

Li, Tianling; Winnel, Melissa; Lin, Hao; Panther, Jared; Liu, Chang; O'Halloran, Roger; Wang, Kewen; An, Taicheng; Wong, Po Keung; Zhang, Shanqing; Zhao, Huijun

2017-09-15

With closing water loop through purified recycled water, wastewater becomes a part of source water, requiring reliable wastewater quality monitoring system (WQMS) to manage wastewater source and mitigate potential health risks. However, the development of reliable WQMS is fatally constrained by severe contamination and biofouling of sensors due to the hostile analytical environment of wastewaters, especially raw sewages, that challenges the limit of existing sensing technologies. In this work, we report a technological solution to enable the development of WQMS for real-time abnormal event detection with high reliability and practicality. A vectored high flow hydrodynamic self-cleaning approach and a dual-sensor self-diagnostic concept are adopted for WQMS to effectively encounter vital sensor failing issues caused by contamination and biofouling and ensure the integrity of sensing data. The performance of the WQMS has been evaluated over a 3-year trial period at different sewage catchment sites across three Australian states. It has demonstrated that the developed WQMS is capable of continuously operating in raw sewage for a prolonged period up to 24 months without maintenance and failure, signifying the high reliability and practicality. The demonstrated WQMS capability to reliably acquire real-time wastewater quality information leaps forward the development of effective wastewater source management system. The reported self-cleaning and self-diagnostic concepts should be applicable to other online water quality monitoring systems, opening a new way to encounter the common reliability and stability issues caused by sensor contamination and biofouling. Copyright © 2017 Elsevier Ltd. All rights reserved.

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.

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

Reliability analysis based on the losses from failures.

Science.gov (United States)

Todinov, M T

2006-04-01

early-life failures region and the expected losses given failure characterizing the corresponding time intervals. For complex systems whose components are not logically arranged in series, discrete simulation algorithms and software have been created for determining the losses from failures in terms of expected lost production time, cost of intervention, and cost of replacement. Different system topologies are assessed to determine the effect of modifications of the system topology on the expected losses from failures. It is argued that the reliability allocation in a production system should be done to maximize the profit/value associated with the system. Consequently, a method for setting reliability requirements and reliability allocation maximizing the profit by minimizing the total cost has been developed. Reliability allocation that maximizes the profit in case of a system consisting of blocks arranged in series is achieved by determining for each block individually the reliabilities of the components in the block that minimize the sum of the capital, operation costs, and the expected losses from failures. A Monte Carlo simulation based net present value (NPV) cash-flow model has also been proposed, which has significant advantages to cash-flow models based on the expected value of the losses from failures per time interval. Unlike these models, the proposed model has the capability to reveal the variation of the NPV due to different number of failures occurring during a specified time interval (e.g., during one year). The model also permits tracking the impact of the distribution pattern of failure occurrences and the time dependence of the losses from failures.

Two-phase flow characteristics in BWRs

International Nuclear Information System (INIS)

Katono, Kenichi; Aoyama, Goro; Nagayoshi, Takuji; Yasuda, Kenichi; Nishida, Koji

2014-01-01

Reliable prediction of two-phase flow characteristics is important for safety and economy improvements of BWR plants. We have been developing two-phase flow measurement tools and techniques for BWR thermal hydraulic conditions, such as a 3D time-averaged X-ray CT system, an ultrasonic liquid film sensor and a wire-mesh sensor. We applied the developed items in experiments using the multi-purpose steam-water test facility known as HUSTLE, which can simulate two-phase thermal-hydraulic conditions in a BWR reactor pressure vessel, and we constructed a detailed instrumentation database. We validated a 3D two-phase flow simulator using the database and developed the reactor internal two-phase flow analysis system. (author)

Multiplex flow cytometry barcoding and antibody arrays identify surface antigen profiles of primary and metastatic colon cancer cell lines.

Directory of Open Access Journals (Sweden)

Kumar Sukhdeo

Full Text Available Colon cancer is a deadly disease affecting millions of people worldwide. Current treatment challenges include management of disease burden as well as improvements in detection and targeting of tumor cells. To identify disease state-specific surface antigen signatures, we combined fluorescent cell barcoding with high-throughput flow cytometric profiling of primary and metastatic colon cancer lines (SW480, SW620, and HCT116. Our multiplexed technique offers improvements over conventional methods by permitting the simultaneous and rapid screening of cancer cells with reduced effort and cost. The method uses a protein-level analysis with commercially available antibodies on live cells with intact epitopes to detect potential tumor-specific targets that can be further investigated for their clinical utility. Multiplexed antibody arrays can easily be applied to other tumor types or pathologies for discovery-based approaches to target identification.

A Flow Cytometry Protocol to Estimate DNA Content in the Yellowtail Tetra Astyanax altiparanae

Directory of Open Access Journals (Sweden)

Pedro L. P. Xavier

2017-09-01

Full Text Available The production of triploid yellowtail tetra Astyanax altiparanae is a key factor to obtain permanently sterile individuals by chromosome set manipulation. Flow cytometric analysis is the main tool for confirmation of the resultant triploids individuals, but very few protocols are specific for A. altiparanae species. The current study has developed a protocol to estimate DNA content in this species. Furthermore, a protocol for long-term storage of dorsal fins used for flow cytometry analysis was established. The combination of five solutions with three detergents (Nonidet P-40 Substitute, Tween 20, and Triton X-100 at 0.1, 0.2, and 0.4% concentration was evaluated. Using the best solution from this first experiment, the addition of trypsin (0.125, 0.25, and 0.5% and sucrose (74 mM and the effects of increased concentrations of the detergents at 0.6 and 1.2% concentration were also evaluated. After adjustment of the protocol for flow cytometry, preservation of somatic tissue or isolated nuclei was also evaluated by freezing (at −20°C and fixation in saturated NaCl solution, acetic methanol (1:3, ethanol, and formalin at 10% for 30 or 60 days of storage at 25°C. Flow cytometry analysis in yellowtail tetra species was optimized using the following conditions: lysis solution: 9.53 mM MgCl2.7H20; 47.67 mM KCl; 15 mM Tris; 74 mM sucrose, 0.6% Triton X-100, pH 8.0; staining solution: Dulbecco's PBS with DAPI 1 μg mL−1; preservation procedure: somatic cells (dorsal fin samples frozen at −20°C. Using this protocol, samples may be stored up to 60 days with good accuracy for flow cytometry analysis.

Estimating the Optimal Capacity for Reservoir Dam based on Reliability Level for Meeting Demands

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Mehrdad Taghian

2017-02-01

Full Text Available Introduction: One of the practical and classic problems in the water resource studies is estimation of the optimal reservoir capacity to satisfy demands. However, full supplying demands for total periods need a very high dam to supply demands during severe drought conditions. That means a major part of reservoir capacity and costs is only usable for a short period of the reservoir lifetime, which would be unjustified in economic analysis. Thus, in the proposed method and model, the full meeting demand is only possible for a percent time of the statistical period that is according to reliability constraint. In the general methods, although this concept apparently seems simple, there is a necessity to add binary variables for meeting or not meeting demands in the linear programming model structures. Thus, with many binary variables, solving the problem will be time consuming and difficult. Another way to solve the problem is the application of the yield model. This model includes some simpler assumptions and that is so difficult to consider details of the water resource system. The applicationof evolutionary algorithms, for the problems have many constraints, is also very complicated. Therefore, this study pursues another solution. Materials and Methods: In this study, for development and improvement the usual methods, instead of mix integer linear programming (MILP and the above methods, a simulation model including flow network linear programming is used coupled with an interface manual code in Matlab to account the reliability based on output file of the simulation model. The acre reservoir simulation program (ARSP has been utilized as a simulation model. A major advantage of the ARSP is its inherent flexibility in defining the operating policies through a penalty structure specified by the user. The ARSP utilizes network flow optimization techniques to handle a subset of general linear programming (LP problems for individual time intervals

Impact of maternal gestational diabetes on neutrophil functions of ...

African Journals Online (AJOL)

Ehab

cell-mediated immunity and lower chemotactic activity of cord blood ... neonates born to gestational diabetic mothers on insulin during pregnancy and another 15 born to healthy ... 2) Flow cytometric assay of oxidative burst using. DHR 123:.

Contributions to the determination of the thermal core reliability of pressurized water reactors

International Nuclear Information System (INIS)

Ackermann, G.; Horche, W.; Melchior, H.; Prasser, H.M.

1982-09-01

The investigations in the field of thermohydraulics of PW reactors are aimed at a possible increase of economy and reliability of WWER-type-reactors. In detail the flow distribution at the core entrance, the modification of the power distribution as a result of an irregular temperature distribution at the core entrance, and based on the theory of hot spots the thermic core reliability are studied. In this connection qualitatively new methods are applied characterized by low expenditure. (author)

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.

and virus-mediated processes for better understanding of the microbial food web and the biogeochemistry. 1. Suttle, C. A., Nature, 2005, 437, 356– 361. 2. Danovaro, R. et al., Freshwater Biol., 2008, 53, 1186–1213. 3. Suttle, C. A., Nature, 2007, 5... of the microbial food web, with abundance in marine waters ranging from 10 6 ml –1 in the deep sea to 10 8 ml –1 in coastal waters and 10 9 g –1 of dry weight in the marine sediments 1,2 , which is usually 15-fold greater than bacterial and archael...

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

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

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.

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...

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)

Analysis of methods to estimate spring flows in a karst aquifer.

Science.gov (United States)

Sepúlveda, Nicasio

2009-01-01

Hydraulically and statistically based methods were analyzed to identify the most reliable method to predict spring flows in a karst aquifer. Measured water levels at nearby observation wells, measured spring pool altitudes, and the distance between observation wells and the spring pool were the parameters used to match measured spring flows. Measured spring flows at six Upper Floridan aquifer springs in central Florida were used to assess the reliability of these methods to predict spring flows. Hydraulically based methods involved the application of the Theis, Hantush-Jacob, and Darcy-Weisbach equations, whereas the statistically based methods were the multiple linear regressions and the technology of artificial neural networks (ANNs). Root mean square errors between measured and predicted spring flows using the Darcy-Weisbach method ranged between 5% and 15% of the measured flows, lower than the 7% to 27% range for the Theis or Hantush-Jacob methods. Flows at all springs were estimated to be turbulent based on the Reynolds number derived from the Darcy-Weisbach equation for conduit flow. The multiple linear regression and the Darcy-Weisbach methods had similar spring flow prediction capabilities. The ANNs provided the lowest residuals between measured and predicted spring flows, ranging from 1.6% to 5.3% of the measured flows. The model prediction efficiency criteria also indicated that the ANNs were the most accurate method predicting spring flows in a karst aquifer.

Measuring T cell-mediated cytotoxicity using fluorogenic caspase substrates.

Science.gov (United States)

Chahroudi, A; Silvestri, G; Feinberg, M B

2003-10-01

Cytotoxic T lymphocytes (CTLs) play a major role in the immune response against viruses and other intracellular pathogens. In addition, CTLs are implicated in the control of tumor cells in certain settings. Accurate measures of CTL function are of critical importance to study the pathogenesis of infectious diseases and to evaluate the efficacy of new vaccines and immunotherapies. To this end, we have recently developed a flow cytometry-based CTL (FCC) assay that measures the CTL-induced caspase activation within target cells using cell permeable fluorogenic caspase substrates. This novel assay reliably detects, by flow cytometry or fluorescence/confocal microscopy, antigen-specific CTLs in a wide variety of human and murine systems, and is safer and more informative than the standard 51Cr-release assay. In addition, the flow cytometric CTL (FCC) assay provides an alternative method that is often more sensitive and physiologically informative when compared to previously described FCC assays, as it measures a biological indicator of apoptosis within the target cell. The FCC assay may thus represent a useful tool to further understand the molecular and cellular mechanisms that underlie CTL-mediated killing during tumorigenesis or following infection with viruses or other intracellular pathogens.

Transmission embedded cost allocation methodology with consideration of system reliability

International Nuclear Information System (INIS)

Hur, D.; Park, J.-K.; Yoo, C.-I.; Kim, B.H.

2004-01-01

In a vertically integrated utility industry, the cost of reliability, as a separate service, has not received much rigorous analysis. However, as a cornerstone of restructuring the industry, the transmission service pricing must change to be consistent with, and supportive of, competitive wholesale electricity markets. This paper focuses on the equitable allocation of transmission network embedded costs including the transmission reliability cost based on the contributions of each generator to branch flows under normal conditions as well as the line outage impact factor under a variety of load levels. A numerical example on a six-bus system is given to illustrate the applications of the proposed methodology. (author)

Provision of reliable core cooling in vessel-type boiling reactors

International Nuclear Information System (INIS)

Alferov, N.S.; Balunov, B.F.; Davydov, S.A.

1987-01-01

Methods for providing reliable core cooling in vessel-type boiling reactors with natural circulation for heat supply are analysed. The solution of this problem is reduced to satisfaction of two conditions such as: water confinement over the reactor core necessary in case of an accident and confinement of sufficient coolant flow rate through the bottom cross section of fuel assemblies for some time. The reliable fuel element cooling under conditions of a maximum credible accident (brittle failure of a reactor vessel) is shown to be provided practically in any accident, using the safety vessel in combination with the application of means of standard operation and minimal composition and capacity of ECCS

Standardization and validation of a cytometric bead assay to assess antibodies to multiple Plasmodium falciparum recombinant antigens

Directory of Open Access Journals (Sweden)

Ondigo Bartholomew N

2012-12-01

Full Text Available Abstract Background Multiplex cytometric bead assay (CBA have a number of advantages over ELISA for antibody testing, but little information is available on standardization and validation of antibody CBA to multiple Plasmodium falciparum antigens. The present study was set to determine optimal parameters for multiplex testing of antibodies to P. falciparum antigens, and to compare results of multiplex CBA to ELISA. Methods Antibodies to ten recombinant P. falciparum antigens were measured by CBA and ELISA in samples from 30 individuals from a malaria endemic area of Kenya and compared to known positive and negative control plasma samples. Optimal antigen amounts, monoplex vs multiplex testing, plasma dilution, optimal buffer, number of beads required were assessed for CBA testing, and results from CBA vs. ELISA testing were compared. Results Optimal amounts for CBA antibody testing differed according to antigen. Results for monoplex CBA testing correlated strongly with multiplex testing for all antigens (r = 0.88-0.99, P values from Conclusion With optimization, CBA may be the preferred method of testing for antibodies to P. falciparum antigens, as CBA can test for antibodies to multiple recombinant antigens from a single plasma sample and produces a greater range of values in positive samples and lower background readings for blank samples than ELISA.

Apomixis is not prevalent in subnival to nival plants of the European Alps

Czech Academy of Sciences Publication Activity Database

Hörandl, E.; Dobeš, C.; Suda, Jan; Vít, Petr; Urfus, Tomáš; Temsch, E. M.; Cosendai, A.-C.; Wagner, J.; Ladinig, U.

2011-01-01

Roč. 108, č. 2 (2011), s. 381-390 ISSN 0305-7364 Institutional research plan: CEZ:AV0Z60050516 Keywords : apomixis * endosperm * flow cytometric seed screen Subject RIV: EF - Botanics Impact factor: 4.030, year: 2011

Use of internal control T-cell populations in the flow cytometric evaluation for T-cell neoplasms.

Science.gov (United States)

Hunt, Alicia M; Shallenberger, Wendy; Ten Eyck, Stephen P; Craig, Fiona E

2016-09-01

Flow cytometry is an important tool for identification of neoplastic T-cells, but immunophenotypic abnormalities are often subtle and must be distinguished from nonneoplastic subsets. Use of internal control (IC) T-cells in the evaluation for T-cell neoplasms was explored, both as a quality measure and as a reference for evaluating abnormal antigen expression. All peripheral blood specimens (3-month period), or those containing abnormal T-cells (29-month period), stained with CD45 V500, CD2 V450, CD3 PE-Cy7, CD7 PE, CD4 Per-CP-Cy5.5, CD8 APC-H7, CD56 APC, CD16&57 FITC, were evaluated. IC T-cells were identified (DIVA, BD Biosciences) and median fluorescence intensity (MFI) recorded. Selected files were merged and reference templates generated (Infinicyt, Cytognos). IC T-cells were present in all specimens, including those with abnormal T-cells, but subsets were less well-represented. IC T-cell CD3 MFI differed between instruments (p = 0.0007) and subsets (p < 0.001), but not specimen categories, and served as a longitudinal process control. Merged files highlighted small unusual IC-T subsets: CD2+(dim) (0.25% total), CD2- (0.03% total). An IC reference template highlighted neoplastic T-cells, but was limited by staining variability (IC CD3 MFI reference samples different from test (p = 0.003)). IC T-cells present in the majority of specimens can serve as positive and longitudinal process controls. Use of IC T-cells as an internal reference is limited by variable representation of subsets. Analysis of merged IC T-cells from previously analyzed patient samples can alert the interpreter to less-well-recognized non-neoplastic subsets. However, application of a merged file IC reference template was limited by staining variability. © 2016 Clinical Cytometry Society. © 2016 International Clinical Cytometry Society.

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

Gastric lymphomas in Turkey. Analysis of prognostic factors with special emphasis on flow cytometric DNA content.

Science.gov (United States)

Aydin, Z D; Barista, I; Canpinar, H; Sungur, A; Tekuzman, G

2000-07-01

In contrast to DNA ploidy, to the authors' knowledge the prognostic significance of S-phase fraction (SPF) in gastric lymphomas has not been determined. In the current study, the prognostic significance of various parameters including SPF and DNA aneuploidy were analyzed and some distinct epidemiologic and biologic features of gastric lymphomas in Turkey were found. A series of 78 gastric lymphoma patients followed at Hacettepe University is reported. DNA flow cytometry was performed for 34 patients. The influence of various parameters on survival was investigated with the log rank test. The Cox proportional hazards model was fitted to identify independent prognostic factors. The median age of the patients was 50 years. There was no correlation between patient age and tumor grade. DNA content analysis revealed 4 of the 34 cases to be aneuploid with DNA index values < 1.0. The mean SPF was 33.5%. In the univariate analysis, surgical resection of the tumor, modified Ann Arbor stage, performance status, response to first-line chemotherapy, lactate dehydrogenase (LDH) level, and SPF were important prognostic factors for disease free survival (DFS). The same parameters, excluding LDH level, were important for determining overall survival (OS). In the multivariate analysis, surgical resection of the tumor, disease stage, performance status, and age were found to be important prognostic factors for OS. To the authors' knowledge the current study is the first to demonstrate the prognostic significance of SPF in gastric lymphomas. The distinguishing features of Turkish gastric lymphoma patients are 1) DNA indices of aneuploid cases that all are < 1.0, which is a unique feature; 2) a lower percentage of aneuploid cases; 3) a higher SPF; 4) a younger age distribution; and 5) lack of an age-grade correlation. The authors conclude that gastric lymphomas in Turkey have distinct biologic and epidemiologic characteristics. Copyright 2000 American Cancer Society.

A water treatment case study for quantifying model performance with multilevel flow modeling

Directory of Open Access Journals (Sweden)

Emil K. Nielsen

2018-05-01

Full Text Available Decision support systems are a key focus of research on developing control rooms to aid operators in making reliable decisions and reducing incidents caused by human errors. For this purpose, models of complex systems can be developed to diagnose causes or consequences for specific alarms. Models applied in safety systems of complex and safety-critical systems require rigorous and reliable model building and testing. Multilevel flow modeling is a qualitative and discrete method for diagnosing faults and has previously only been validated by subjective and qualitative means. To ensure reliability during operation, this work aims to synthesize a procedure to measure model performance according to diagnostic requirements. A simple procedure is proposed for validating and evaluating the concept of multilevel flow modeling. For this purpose, expert statements, dynamic process simulations, and pilot plant experiments are used for validation of simple multilevel flow modeling models of a hydrocyclone unit for oil removal from produced water. Keywords: Fault Diagnosis, Model Validation, Multilevel Flow Modeling, Produced Water Treatment

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.

Multinomial-exponential reliability function: a software reliability model

International Nuclear Information System (INIS)

Saiz de Bustamante, Amalio; Saiz de Bustamante, Barbara

2003-01-01

The multinomial-exponential reliability function (MERF) was developed during a detailed study of the software failure/correction processes. Later on MERF was approximated by a much simpler exponential reliability function (EARF), which keeps most of MERF mathematical properties, so the two functions together makes up a single reliability model. The reliability model MERF/EARF considers the software failure process as a non-homogeneous Poisson process (NHPP), and the repair (correction) process, a multinomial distribution. The model supposes that both processes are statistically independent. The paper discusses the model's theoretical basis, its mathematical properties and its application to software reliability. Nevertheless it is foreseen model applications to inspection and maintenance of physical systems. The paper includes a complete numerical example of the model application to a software reliability analysis

Surface roughness influences on the behaviour of flow inside microchannels

Science.gov (United States)

Farias, M. H.; Castro, C. S.; Garcia, D. A.; Henrique, J. S.

2018-03-01

This work discusses influence of the surface roughness on the behavior of liquids flowing inside microchannels. By measuring the flow profile using the micro-PIV technique, the flow of water inside two rectangular microchannels of different wall roughness and in a circular smooth microchannel was studied. Comparisons were made among the experimental results, showing that a metrological approach concerning surface characteristics of microdevices is required to ensure reliability of the measurements for flow analyses in microfluidic processes.

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.

Method for screening inhibitors of the toxicity of Bacillus anthracis

Science.gov (United States)

Cirino, Nick M.; Jackson, Paul J.; Lehnert, Bruce E.

2001-01-01

The protective antigen (PA) of Bacillus anthracis is integral to the mechanism of anthrax poisoning. The cloning, expression and purification of a 32 kDa B. anthracis PA fragment (PA32) is described. This fragment has also been expressed as a fusion construct to stabilized green fluorescent protein (EGFP-PA32). Both proteins were capable of binding to specific cell surface receptors as determined by fluorescent microscopy and a flow cytometric assay. To confirm binding specificity in the flow cytometric assay, non-fluorescent PA83 or PA32 was used to competitively inhibit fluorescent EGFP-PA32 binding to cell receptors. This assay can be employed as a rapid screen for compounds which disrupts binding of PA to cells. Additionally, the high intracellular expression levels and ease of purification make this recombinant protein an attractive vaccine candidate or therapeutic treatment for anthrax poisoning.

reliability reliability

African Journals Online (AJOL)

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Corresponding author, Tel: +234-703. RELIABILITY .... V , , given by the code of practice. However, checks must .... an optimization procedure over the failure domain F corresponding .... of Concrete Members based on Utility Theory,. Technical ...

Measuring reliability under epistemic uncertainty: Review on non-probabilistic reliability metrics

Directory of Open Access Journals (Sweden)

Kang Rui

2016-06-01

Full Text Available In this paper, a systematic review of non-probabilistic reliability metrics is conducted to assist the selection of appropriate reliability metrics to model the influence of epistemic uncertainty. Five frequently used non-probabilistic reliability metrics are critically reviewed, i.e., evidence-theory-based reliability metrics, interval-analysis-based reliability metrics, fuzzy-interval-analysis-based reliability metrics, possibility-theory-based reliability metrics (posbist reliability and uncertainty-theory-based reliability metrics (belief reliability. It is pointed out that a qualified reliability metric that is able to consider the effect of epistemic uncertainty needs to (1 compensate the conservatism in the estimations of the component-level reliability metrics caused by epistemic uncertainty, and (2 satisfy the duality axiom, otherwise it might lead to paradoxical and confusing results in engineering applications. The five commonly used non-probabilistic reliability metrics are compared in terms of these two properties, and the comparison can serve as a basis for the selection of the appropriate reliability metrics.

Agonist-induced platelet reactivity correlates with bleeding in haemato-oncological patients

NARCIS (Netherlands)

Batman, B.; van Bladel, E. R.; van Hamersveld, M.; Pasker-De Jong, Pieternel C M; Korporaal, S. J.A.; Urbanus, R. T.; Roest, M.; Boven, Leonie A; Fijnheer, R.

2017-01-01

Background and objective: Prophylactic platelet transfusions are administered to prevent bleeding in haemato-oncological patients. However, bleeding still occurs, despite these transfusions. This practice is costly and not without risk. Better predictors of bleeding are needed, and flow cytometric

Rapid MR measurements of contrast medium dilution kinetics (gadolinium-DTPA) in a flow phantom

International Nuclear Information System (INIS)

Boeck, J.C.; Sander, B.; Frank, J.; Schoerner, W.

1991-01-01

We studied first-pass MRI-contrast dilution to compute flow and volume of distribution in a realistic flow phantom. Pulsatile flow was provided by a one-chamber artificial heart. Physiological stroke volume, rate, pressure, and flow were adjustable. An elastic tube with dimensions similar to that of the human aorta was imaged at a rate of 2.4 Hz. After contrast injection, an initial increase in signal intensity was followed by a decrease. Signal-intensity-time plots demonstrated slightly skewed curves as expected from dispersion theory. After calibration at different gadolinium-DTPA concentrations, signal intensities were converted into true gadolinium concentrations, and flow was calculated from the concentration-time curves. Flow was varied between 2.5 and 10.0 l/min and a significant correlation was found between the MRI-estimate and true flow. Volume of distribution between injection and detection site was reliably estimated. This study demonstrates rapid 2-D imaging of a paramagnetic contrast bolus in a realistic flow phantom. Reliable estimates of flow and volume are obtained. (orig.) [de

Calibration measurements using the ORNL fissile mass flow monitor

International Nuclear Information System (INIS)

March-Leuba, J.; Uckan, T.; Sumner, J.; Mattingly, J.; Mihalczo, J.

1998-01-01

This paper presents a demonstration of fissile-mass-flow measurements using the Oak Ridge National Laboratory (ORNL) Fissile Mass Flow Monitor in the Paducah Gaseous Diffusion Plant (PGDP). This Flow Monitor is part of a Blend Down Monitoring System (BDMS) that will be installed in at least two Russian Federation (R.F.) blending facilities. The key objectives of the demonstration of the ORNL Flow Monitor are two: (a) demonstrate that the ORNL Flow Monitor equipment is capable of reliably monitoring the mass flow rate of 235 UF 6 gas, and (b) provide a demonstration of ORNL Flow Monitor system in operation with UF 6 flow for a visiting R.F. delegation. These two objectives have been met by the PGDP demonstration, as presented in this paper

The utility of flow cytometry in differentiating NK/T cell lymphoma from indolent and reactive NK cell proliferations.

Science.gov (United States)

de Mel, Sanjay; Li, Jenny Bei; Abid, Muhammad Bilal; Tang, Tiffany; Tay, Hui Ming; Ting, Wen Chang; Poon, Li Mei; Chung, Tae Hoon; Mow, Benjamin; Tso, Allison; Ong, Kiat Hoe; Chng, Wee Joo; Liu, Te Chih

2018-01-01

The WHO defines three categories of NK cell malignancies; extra nodal NK/T cell lymphoma (NKTCL), aggressive NK cell leukemia, and the provisional entity chronic lymphoproliferative disorder of NK cells (CLPD-NK). Although the flow cytometric (FC) phenotype of CLPD-NK has been described, studies on FC phenotype of NKTCL are limited. To the best of our knowledge ours is the first study to compare the phenotype of NKTCL, CLPD-NK, reactive NK lymphocytosis (RNKL), and normal NK cells using eight color (8C) FC. Specimens analyzed using the Euroflow8C NK Lymphoproliferative Disorder (NKLPD) panel between 2011 and 2014 were identified from our database. All samples were analyzed on the FACSCantoII cytometer. NK cells were identified as CD45+, smCD3-, CD19-, CD56+ and normal T-cells served as internal controls. The majority of NKTCL were CD56 bright, CD16 dim, CD57-, and CD94+. CLPD-NK and RNKL were predominantly CD56+ or dim with positive expression of CD16 and CD57 and weak CD94 expression. Antigen based statistical analyses showed robust division of samples along the NKTCL/normal CD56 bright NK cell and CLPD-NK/RNKL/normal CD56 positive NK cell groups. It was concluded that FC can reliably distinguish NKTCL from CLPD-NK, normal NK cells of CD56+ phenotype, and RNKL. It was proposed that the typical phenotype for NKTCL is: CD56 bright, CD16 dim with positive CD2, CD7, CD94, HLADR, CD25, CD26, and absent CD57. This resembles the phenotype of the CD56 bright immunoregulatory subset of NK cells which we therefore hypothesize is the cell of origin of NKTCL. © 2017 International Clinical Cytometry Society. © 2017 International Clinical Cytometry Society.

An application of the fault tree analysis for the power system reliability estimation

International Nuclear Information System (INIS)

Volkanovski, A.; Cepin, M.; Mavko, B.

2007-01-01

The power system is a complex system with its main function to produce, transfer and provide consumers with electrical energy. Combinations of failures of components in the system can result in a failure of power delivery to certain load points and in some cases in a full blackout of power system. The power system reliability directly affects safe and reliable operation of nuclear power plants because the loss of offsite power is a significant contributor to the core damage frequency in probabilistic safety assessments of nuclear power plants. The method, which is based on the integration of the fault tree analysis with the analysis of the power flows in the power system, was developed and implemented for power system reliability assessment. The main contributors to the power system reliability are identified, both quantitatively and qualitatively. (author)

Design flow factors for sewerage systems in small arid communities

OpenAIRE

Imam, Emad H.; Elnakar, Haitham Y.

2014-01-01

Reliable estimation of sewage flow rates is essential for the proper design of sewers, pumping stations, and treatment plants. The design of the various components of the sewerage system should be based on the most critical flow rates with a focus on extremely low and peak flow rates that would be sustained for a duration related to the acceptable limits of behavior of the components under consideration. The extreme flow conditions and to what extent they differ from the average values are cl...

Frontiers of reliability

CERN Document Server

Basu, Asit P; Basu, Sujit K

1998-01-01

This volume presents recent results in reliability theory by leading experts in the world. It will prove valuable for researchers, and users of reliability theory. It consists of refereed invited papers on a broad spectrum of topics in reliability. The subjects covered include Bayesian reliability, Bayesian reliability modeling, confounding in a series system, DF tests, Edgeworth approximation to reliability, estimation under random censoring, fault tree reduction for reliability, inference about changes in hazard rates, information theory and reliability, mixture experiment, mixture of Weibul

An exact method for solving logical loops in reliability analysis

International Nuclear Information System (INIS)

Matsuoka, Takeshi

2009-01-01

This paper presents an exact method for solving logical loops in reliability analysis. The systems that include logical loops are usually described by simultaneous Boolean equations. First, present a basic rule of solving simultaneous Boolean equations. Next, show the analysis procedures for three-component system with external supports. Third, more detailed discussions are given for the establishment of logical loop relation. Finally, take up two typical structures which include more than one logical loop. Their analysis results and corresponding GO-FLOW charts are given. The proposed analytical method is applicable to loop structures that can be described by simultaneous Boolean equations, and it is very useful in evaluating the reliability of complex engineering systems.

Flow-Control Unit For Nitrogen And Hydrogen Gases

Science.gov (United States)

Chang, B. J.; Novak, D. W.

1990-01-01

Gas-flow-control unit installed and removed as one piece replaces system that included nine separately serviced components. Unit controls and monitors flows of nitrogen and hydrogen gases. Designed for connection via fluid-interface manifold plate, reducing number of mechanical fluid-interface connections from 18 to 1. Unit provides increasing reliability, safety, and ease of maintenance, and for reducing weight, volume, and power consumption.

Software reliability

CERN Document Server

Bendell, A

1986-01-01

Software Reliability reviews some fundamental issues of software reliability as well as the techniques, models, and metrics used to predict the reliability of software. Topics covered include fault avoidance, fault removal, and fault tolerance, along with statistical methods for the objective assessment of predictive accuracy. Development cost models and life-cycle cost models are also discussed. This book is divided into eight sections and begins with a chapter on adaptive modeling used to predict software reliability, followed by a discussion on failure rate in software reliability growth mo

Enhancing chemical synthesis using catalytic reactions under continuous flow conditions

OpenAIRE

Asadi, Mousa

2017-01-01

Many advantages have been demonstrated for continuous flow chemistry in comparison with batch chemistry; such as easy automation, high level of reproducibility, improved safety, and process reliability. Indeed, with continuous flow processes constant reaction parameters such as temperature, time, amount of reagents, catalyst, solvents, efficient mixing etc. can easily be assured. The research detailed in this PhD thesis takes advantages of flow chemistry applying it to the Fukuyama ...

Studies of thermal-hydrodynamic flow instability, 2

International Nuclear Information System (INIS)

Suzuoki, Akira

1977-01-01

For reliable prediction of flow stability in sodium-heated steam generators, a dynamic model was proposed for boiling flow oscillation in parallel channel systems, and an analysis code was developed. The model contains a description of a sodium flow exchanging heat with a water flow in counter-current fashion. The code was applied to three representative flow systems whose heating conditions differed from each other, whereby their flow stabilities were compared with a focus on the effects of heating condition. Eigenvalues and flow impedances of the oscillation determined for each system reveal that: (1) Two fundamental systems for the steam generator, parallel tube system in an evaporator and steam generator modules arranged in parallel, have different stabilities under low frequency oscillation. (2) Existing analysis model conditioned on constant heat flux gives different results on stability from those of either steam generator model under low frequency oscillation. (auth.)

Single cell analysis of gene expression patterns of competence development and initiation of sporulation in Bacillus subtilis grown on chemically defined media

NARCIS (Netherlands)

Veening, J. -W.; Smits, W. K.; Hamoen, L. W.; Kuipers, O. P.

Aim: Understanding the basis for the heterogeneous (or bistable) expression patterns of competence development and sporulation in Bacillus subtilis. Methods and Results: Using flow cytometric analyses of various promoter-GFP fusions, we have determined the single-cell gene expression patterns of

Head kidney-derived macrophages of common carp (Cyprinus carpio L.) show plasticity and functional polarization upon differential stimulation

NARCIS (Netherlands)

Joerink, Maaike; Ribeiro, Carla M. S.; Stet, René J. M.; Hermsen, Trudi; Savelkoul, Huub F. J.; Wiegertjes, Geert F.

2006-01-01

Cells from the myeloid lineage are pluripotent. To investigate the potential of myeloid cell polarization in a primitive vertebrate species, we phenotypically and functionally characterized myeloid cells of common carp (Cyprinus carpio L.) during culture. Flow cytometric analysis, Ab labeling of

System Reliability Engineering

International Nuclear Information System (INIS)

Lim, Tae Jin

2005-02-01

This book tells of reliability engineering, which includes quality and reliability, reliability data, importance of reliability engineering, reliability and measure, the poisson process like goodness of fit test and the poisson arrival model, reliability estimation like exponential distribution, reliability of systems, availability, preventive maintenance such as replacement policies, minimal repair policy, shock models, spares, group maintenance and periodic inspection, analysis of common cause failure, and analysis model of repair effect.

Flow karyotyping and flow instrumentation development. Final report

Energy Technology Data Exchange (ETDEWEB)

GEngh, G.J. van den

1997-11-01

The project had three major aims: improvement of technology for high-speed cell and chromosome sorting; the use of such instrumentation in genome analysis; applying the principles developed and the lessons learned to automated processes for the genome program. The work was a continuation of studies that were started at the Lawrence Livermore National Laboratory before the PI moved to the University of Washington. At Livermore, a high-speed sorter for the selection of human chromosomes was built. The instrument incorporated Livermore`s advanced sorter technology. The engineering focused on improving robustness and reliability so that the full potential of high-speed sorting would become available to the biological research laboratory. The new instrument, dubbed MoFlo for modular flow cytometer, proved to be a very practical and efficient tool during the chromosome isolation phase of the gene-library project. Its reliability and ease of operation exceeded that of the commercial instruments. The technology was licensed to two companies.

Fertility and flow cytometric evaluations of frozen-thawed rooster semen in cryopreservation medium containing low-density lipoprotein.

Science.gov (United States)

Shahverdi, A; Sharafi, M; Gourabi, H; Yekta, A Amiri; Esmaeili, V; Sharbatoghli, M; Janzamin, E; Hajnasrollahi, M; Mostafayi, F

2015-01-01

Frozen-thawed rooster semen is not reliable for use in artificial insemination in commercial stocks. Low-density lipoprotein (LDL) has been assessed for effectiveness as a cryoprotectant in the extender to improve the quality of frozen-thawed rooster semen. Although LDL has been evaluated in a few studies in other species for semen cryopreservation, so far no study has been conducted to examine this cryoprotectant for cryopreservation of fowl semen. Thus, this study aims to analyze the effects of different concentrations of LDL (0%, 2%, 4%, 6%, and 8%) in a Beltsville extender for cryopreservation of rooster spermatozoa. In experiment 1, motion parameters, membrane integrity, acrosome integrity, apoptosis status, and mitochondria activity were assessed after freeze-thawing. The highest quality frozen-thawed semen was selected to be used for evaluation of the fertility rate in experiment 2. Semen was collected from six roosters, twice weekly, then extended in a Beltsville extender that contained different concentrations of LDL as follows: 0% (control), 1% (Beltsville plus 1% LDL [BLDL1]), 2% (BLDL2), 4% (BLDL4), 6% (BLDL6), and 8% (BLDL8). Supplementation of the Beltsville extender with 4% LDL produced the most significant percentage of motility (43.1 ± 1.3), membrane integrity (59.4 ± 2.1),mitochondria activity (49.1 ± 1.19), and viable spermatozoa (45 ± 2.28) compared with the control treatment with the results of 22.7 ± 1.3 (motility), 38.4 ± 2.1 (membrane integrity), 40.25 ± 1.19 (mitochondrial activity), and 37.8 ± 2.28 (viability). In experiment 2, a significantly higher percentage of fertility rate was observed for frozen-thawed semen in the extender supplemented with 4% LDL (49.5 ± 1.6) compared with the control (29.2 ± 2.9). Progressive motility and acrosome integrity were not affected by LDL levels in the extenders. The results revealed that supplementation of the Beltsville extender with 4% LDL resulted in higher quality of frozen-thawed rooster

A reliability simulation language for reliability analysis

International Nuclear Information System (INIS)

Deans, N.D.; Miller, A.J.; Mann, D.P.

1986-01-01

The results of work being undertaken to develop a Reliability Description Language (RDL) which will enable reliability analysts to describe complex reliability problems in a simple, clear and unambiguous way are described. Component and system features can be stated in a formal manner and subsequently used, along with control statements to form a structured program. The program can be compiled and executed on a general-purpose computer or special-purpose simulator. (DG)

A study of operational and testing reliability in software reliability analysis

International Nuclear Information System (INIS)

Yang, B.; Xie, M.

2000-01-01

Software reliability is an important aspect of any complex equipment today. Software reliability is usually estimated based on reliability models such as nonhomogeneous Poisson process (NHPP) models. Software systems are improving in testing phase, while it normally does not change in operational phase. Depending on whether the reliability is to be predicted for testing phase or operation phase, different measure should be used. In this paper, two different reliability concepts, namely, the operational reliability and the testing reliability, are clarified and studied in detail. These concepts have been mixed up or even misused in some existing literature. Using different reliability concept will lead to different reliability values obtained and it will further lead to different reliability-based decisions made. The difference of the estimated reliabilities is studied and the effect on the optimal release time is investigated

Human reliability

International Nuclear Information System (INIS)

Embrey, D.E.

1987-01-01

Concepts and techniques of human reliability have been developed and are used mostly in probabilistic risk assessment. For this, the major application of human reliability assessment has been to identify the human errors which have a significant effect on the overall safety of the system and to quantify the probability of their occurrence. Some of the major issues within human reliability studies are reviewed and it is shown how these are applied to the assessment of human failures in systems. This is done under the following headings; models of human performance used in human reliability assessment, the nature of human error, classification of errors in man-machine systems, practical aspects, human reliability modelling in complex situations, quantification and examination of human reliability, judgement based approaches, holistic techniques and decision analytic approaches. (UK)

Methods for estimating flow-duration and annual mean-flow statistics for ungaged streams in Oklahoma

Science.gov (United States)

Esralew, Rachel A.; Smith, S. Jerrod

2010-01-01

Flow statistics can be used to provide decision makers with surface-water information needed for activities such as water-supply permitting, flow regulation, and other water rights issues. Flow statistics could be needed at any location along a stream. Most often, streamflow statistics are needed at ungaged sites, where no flow data are available to compute the statistics. Methods are presented in this report for estimating flow-duration and annual mean-flow statistics for ungaged streams in Oklahoma. Flow statistics included the (1) annual (period of record), (2) seasonal (summer-autumn and winter-spring), and (3) 12 monthly duration statistics, including the 20th, 50th, 80th, 90th, and 95th percentile flow exceedances, and the annual mean-flow (mean of daily flows for the period of record). Flow statistics were calculated from daily streamflow information collected from 235 streamflow-gaging stations throughout Oklahoma and areas in adjacent states. A drainage-area ratio method is the preferred method for estimating flow statistics at an ungaged location that is on a stream near a gage. The method generally is reliable only if the drainage-area ratio of the two sites is between 0.5 and 1.5. Regression equations that relate flow statistics to drainage-basin characteristics were developed for the purpose of estimating selected flow-duration and annual mean-flow statistics for ungaged streams that are not near gaging stations on the same stream. Regression equations were developed from flow statistics and drainage-basin characteristics for 113 unregulated gaging stations. Separate regression equations were developed by using U.S. Geological Survey streamflow-gaging stations in regions with similar drainage-basin characteristics. These equations can increase the accuracy of regression equations used for estimating flow-duration and annual mean-flow statistics at ungaged stream locations in Oklahoma. Streamflow-gaging stations were grouped by selected drainage

Flow cytometric analysis reveals the high levels of platelet activation parameters in circulation of multiple sclerosis patients.

Science.gov (United States)

Morel, Agnieszka; Rywaniak, Joanna; Bijak, Michał; Miller, Elżbieta; Niwald, Marta; Saluk, Joanna

2017-06-01

The epidemiological studies confirm an increased risk of cardiovascular disease in multiple sclerosis, especially prothrombotic events directly associated with abnormal platelet activity. The aim of our study was to investigate the level of blood platelet activation in the circulation of patients with chronic phase of multiple sclerosis (SP MS) and their reactivity in response to typical platelets' physiological agonists. We examined 85 SP MS patients diagnosed according to the revised McDonald's criteria and 50 healthy volunteers as a control group. The platelet activation and reactivity were assessed using flow cytometry analysis of the following: P-selectin expression (CD62P), activation of GP IIb/IIIa complex (PAC-1 binding), and formation of platelet microparticles (PMPs) and platelet aggregates (PA) in agonist-stimulated (ADP, collagen) and unstimulated whole blood samples. Furthermore, we measured the level of soluble P-selectin (sP-selectin) in plasma using ELISA method, to evaluate the in vivo level of platelet activation, both in healthy and SP MS subjects. We found a statistically significant increase in P-selectin expression, GP IIb/IIIa activation, and formation of PMPs and PA, as well as in unstimulated and agonist-stimulated (ADP, collagen) platelets in whole blood samples from patients with SP MS in comparison to the control group. We also determined the higher sP-selectin level in plasma of SP MS subjects than in the control group. Based on the obtained results, we might conclude that during the course of SP MS platelets are chronically activated and display hyperreactivity to physiological agonists, such as ADP or collagen.

The Combination of Micro Diaphragm Pumps and Flow Sensors for Single Stroke Based Liquid Flow Control.

Science.gov (United States)

Jenke, Christoph; Pallejà Rubio, Jaume; Kibler, Sebastian; Häfner, Johannes; Richter, Martin; Kutter, Christoph

2017-04-03

With the combination of micropumps and flow sensors, highly accurate and secure closed-loop controlled micro dosing systems for liquids are possible. Implementing a single stroke based control mode with piezoelectrically driven micro diaphragm pumps can provide a solution for dosing of volumes down to nanoliters or variable average flow rates in the range of nL/min to μL/min. However, sensor technologies feature a yet undetermined accuracy for measuring highly pulsatile micropump flow. Two miniaturizable in-line sensor types providing electrical readout-differential pressure based flow sensors and thermal calorimetric flow sensors-are evaluated for their suitability of combining them with mircopumps. Single stroke based calibration of the sensors was carried out with a new method, comparing displacement volumes and sensor flow volumes. Limitations of accuracy and performance for single stroke based flow control are described. Results showed that besides particle robustness of sensors, controlling resistive and capacitive damping are key aspects for setting up reproducible and reliable liquid dosing systems. Depending on the required average flow or defined volume, dosing systems with an accuracy of better than 5% for the differential pressure based sensor and better than 6.5% for the thermal calorimeter were achieved.

Reliability Calculations

DEFF Research Database (Denmark)

Petersen, Kurt Erling

1986-01-01

Risk and reliability analysis is increasingly being used in evaluations of plant safety and plant reliability. The analysis can be performed either during the design process or during the operation time, with the purpose to improve the safety or the reliability. Due to plant complexity and safety...... and availability requirements, sophisticated tools, which are flexible and efficient, are needed. Such tools have been developed in the last 20 years and they have to be continuously refined to meet the growing requirements. Two different areas of application were analysed. In structural reliability probabilistic...... approaches have been introduced in some cases for the calculation of the reliability of structures or components. A new computer program has been developed based upon numerical integration in several variables. In systems reliability Monte Carlo simulation programs are used especially in analysis of very...

Determination of reliable environmental flows in Colombia: The example of the River Palace (Cauca)

International Nuclear Information System (INIS)

Diez Hernandez, Juan Manuel; Ruiz Cobo Dario Hernan

2007-01-01

The increasing exploitation of the water resources in Colombia requires an advanced Environmental Flows Regime (EFR) regulation, for a properly design of the Watershed Planning and Management Programs. Among the diversity of approaches proposed to EFR assessment, the most used and scientifically accepted is the well-known Instream Flow Incremental Methodology (IFIM), whose applicability for Colombian rivers is explored in this study. The IFIM modeling of the Palace River below the diversion of 500 l/s to the new water-supply facilities in Popayan (Cauca) reveals that the global effect of this limited derivation of the 6.78% mean annual flow in the integrity of the aquatic ecosystem is very low. The eco-hydraulic and eco hydrological simulations of the representative reach (500m length, 18m width, 500/00 slope and coarse substrate) analyzed with the software RHABSIM 3.0, revealed some not very significant reductions of the usable habitat for adult fishes and macro-invertebrates. Consequently, the present instream flows regime of the Palace River caused by the diversion project is corroborated as a satisfactory EFR, according to the IFIM evaluation system. Finally, some research lines are suggested, that are focused to the improvement and adaptation of the IFIM to the particular fluvial conditions of Colombia

In vivo analysis of physiological 3D blood flow of cerebral veins

Energy Technology Data Exchange (ETDEWEB)

Schuchardt, Florian; Schroeder, Laure; Baeuerle, Jochen; Harloff, Andreas [University Medical Centre, Department of Neurology, Freiburg (Germany); Anastasopoulos, Constantin [University Medical Center, Department of Neuropaediatrics and Muscle Disorders, Freiburg (Germany); University Medical Centre, Department of Neuroradiology, Freiburg (Germany); Markl, Michael [Northwestern University, Department of Radiology, Feinberg School of Medicine and McCormick School of Engineering, Chicago, IL (United States); Hennemuth, Anja; Drexl, Johann [Fraunhofer MEVIS, Bremen (Germany); Valdueza, Jose M. [Neurological Center, Segeberger Kliniken, Bad Segeberg (Germany); Mader, Irina [University Medical Centre, Department of Neuroradiology, Freiburg (Germany)

2015-08-15

To visualize and quantify physiological blood flow of intracranial veins in vivo using time-resolved, 3D phase-contrast MRI (4D flow MRI), and to test measurement accuracy. Fifteen healthy volunteers underwent repeated ECG-triggered 4D flow MRI (3 Tesla, 32-channel head coil). Intracranial venous blood flow was analysed using dedicated software allowing for blood flow visualization and quantification in analysis planes at the superior sagittal, straight, and transverse sinuses. MRI was evaluated for intra- and inter-observer agreement and scan-rescan reproducibility. Measurements of the transverse sinuses were compared with transcranial two-dimensional duplex ultrasound. Visualization of 3D blood flow within cerebral sinuses was feasible in 100 % and within at least one deep cerebral vein in 87 % of the volunteers. Blood flow velocity/volume increased along the superior sagittal sinus and was lower in the left compared to the right transverse sinus. Intra- and inter-observer reliability and reproducibility of blood flow velocity (mean difference 0.01/0.02/0.02 m/s) and volume (mean difference 0.0002/-0.0003/0.00003 l/s) were good to excellent. High/low velocities were more pronounced (8 % overestimation/9 % underestimation) in MRI compared to ultrasound. Four-dimensional flow MRI reliably visualizes and quantifies three-dimensional cerebral venous blood flow in vivo and is promising for studies in patients with sinus thrombosis and related diseases. (orig.)

Genome-size Variation in Switchgrass (Panicum virgatum: Flow Cytometry and Cytology Reveal Rampant Aneuploidy

Directory of Open Access Journals (Sweden)

Denise E. Costich

2010-11-01

Full Text Available Switchgrass ( 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 considerable variation in genome size within each accession, particularly at the octoploid (2 = 8 = 72 chromosome ploidy level. Highly variable chromosome counts in mitotic cell preparations indicated that aneuploidy was more common in octoploids (86.3% than tetraploids (23.2%. Furthermore, the incidence of hyper- versus hypoaneuploidy is equivalent in tetraploids. This is clearly not the case in octoploids, where close to 90% of the aneuploid counts are lower than the euploid number. Cytogenetic investigation using fluorescent in situ hybridization (FISH revealed an unexpected degree of variation in chromosome structure underlying the apparent genomic instability of this species. These results indicate that rapid advances in the breeding of polyploid biofuel feedstocks, based on the molecular-genetic dissection of biomass characteristics and yield, will be predicated on the continual improvement of our understanding of the cytogenetics of these species.

Minimal disease detection of B-cell lymphoproliferative disorders by flow cytometry: multidimensional cluster analysis.

Science.gov (United States)

Duque, Ricardo E

2012-04-01

Flow cytometric analysis of cell suspensions involves the sequential 'registration' of intrinsic and extrinsic parameters of thousands of cells in list mode files. Thus, it is almost irresistible to describe phenomena in numerical terms or by 'ratios' that have the appearance of 'accuracy' due to the presence of numbers obtained from thousands of cells. The concepts involved in the detection and characterization of B cell lymphoproliferative processes are revisited in this paper by identifying parameters that, when analyzed appropriately, are both necessary and sufficient. The neoplastic process (cluster) can be visualized easily because the parameters that distinguish it form a cluster in multidimensional space that is unique and distinguishable from neighboring clusters that are not of diagnostic interest but serve to provide a background. For B cell neoplasia it is operationally necessary to identify the multidimensional space occupied by a cluster whose kappa:lambda ratio is 100:0 or 0:100. Thus, the concept of kappa:lambda ratio is without meaning and would not detect B cell neoplasia in an unacceptably high number of cases.

Low reproducibility of maximum urinary flow rate determined by portable flowmetry

NARCIS (Netherlands)

Sonke, G. S.; Kiemeney, L. A.; Verbeek, A. L.; Kortmann, B. B.; Debruyne, F. M.; de la Rosette, J. J.

1999-01-01

To evaluate the reproducibility in maximum urinary flow rate (Qmax) in men with lower urinary tract symptoms (LUTSs) and to determine the number of flows needed to obtain a specified reliability in mean Qmax, 212 patients with LUTSs (mean age, 62 years) referred to the University Hospital Nijmegen,

Prediction of Tensile Behavior of UHSFRC Considering the Flow Field in the Placing Dominated by Shear Flow.

Science.gov (United States)

Moon, Joon-Shik; Kang, Su-Tae

2018-01-26

Considering the case of fabricating a UHSFRC (ultra-high strength fiber-reinforced concrete) beam with the method of one end placing and self-flowing to the other end, it was intended to simulate the variation of the fiber orientation distribution according to the flow distance and the variation of the resultant tensile behaviors. Then the validity of the simulation approach was shown by comparing the simulated results with experimental ones. A three-point bending test with a notched beam was adopted for the experiment and a finite element analysis was performed to obtain the simulated results for the bending test considering the flow-dependent tensile behavior of the UHSFRC. From the simulation for the fiber orientation distribution according to the flow distance, it could be found that the major change in the fiber orientation distribution took place within a short flow distance and most of the fibers became nearly aligned to the flow direction. After some flow distance, there was a not-so-remarkable variation in the fiber orientation distribution that could influence the tensile behavior of the composite. For this flow region, the consistent flexural test results, regardless of flow distance, demonstrate the reliability of the simulation.

Reliability in the utility computing era: Towards reliable Fog computing

DEFF Research Database (Denmark)

Madsen, Henrik; Burtschy, Bernard; Albeanu, G.

2013-01-01

This paper considers current paradigms in computing and outlines the most important aspects concerning their reliability. The Fog computing paradigm as a non-trivial extension of the Cloud is considered and the reliability of the networks of smart devices are discussed. Combining the reliability...... requirements of grid and cloud paradigms with the reliability requirements of networks of sensor and actuators it follows that designing a reliable Fog computing platform is feasible....

Fluorescent Cell Barcoding as a Tool to Assess the Age-Related Development of Intracellular Cytokine Production in Small Amounts of Blood from Infants

NARCIS (Netherlands)

Stam, Jose; Abdulahad, Wayel; Huitema, Minke G.; Roozendaal, Caroline; Limburg, Pieter C.; van Stuijvenberg, Margriet; Scholvinck, Elisabeth H.

2011-01-01

Fluorescent Cell Barcoding (FCB) is a flow cytometric technique which has been used for assessing signaling proteins. This FCB technique has the potential to be applied in other multiparameter analyses. Since data on antigen (Ag)-specific T-cell immune responses, like intracellular cytokine

Changes in DNa and microtubules during loss and re-establishment of desiccation tolerance in germinating Medicago truncatula seeds

NARCIS (Netherlands)

Faria, J.M.R.; Buitink, J.; Lammeren, van A.A.M.; Hilhorst, H.W.M.

2005-01-01

Desiccation tolerance (DT) in orthodox seeds is acquired during seed development and lost upon imbibition/germination, purportedly upon the resumption of DNA synthesis in the radicle cells. In the present study, flow cytometric analyses and visualization of microtubules (MTs) in radicle cells of

Dependent systems reliability estimation by structural reliability approach

DEFF Research Database (Denmark)

Kostandyan, Erik; Sørensen, John Dalsgaard

2014-01-01

Estimation of system reliability by classical system reliability methods generally assumes that the components are statistically independent, thus limiting its applicability in many practical situations. A method is proposed for estimation of the system reliability with dependent components, where...... the leading failure mechanism(s) is described by physics of failure model(s). The proposed method is based on structural reliability techniques and accounts for both statistical and failure effect correlations. It is assumed that failure of any component is due to increasing damage (fatigue phenomena...... identification. Application of the proposed method can be found in many real world systems....

Analyzing the reliability of shuffle-exchange networks using reliability block diagrams

International Nuclear Information System (INIS)

Bistouni, Fathollah; Jahanshahi, Mohsen

2014-01-01

Supercomputers and multi-processor systems are comprised of thousands of processors that need to communicate in an efficient way. One reasonable solution would be the utilization of multistage interconnection networks (MINs), where the challenge is to analyze the reliability of such networks. One of the methods to increase the reliability and fault-tolerance of the MINs is use of various switching stages. Therefore, recently, the reliability of one of the most common MINs namely shuffle-exchange network (SEN) has been evaluated through the investigation on the impact of increasing the number of switching stage. Also, it is concluded that the reliability of SEN with one additional stage (SEN+) is better than SEN or SEN with two additional stages (SEN+2), even so, the reliability of SEN is better compared to SEN with two additional stages (SEN+2). Here we re-evaluate the reliability of these networks where the results of the terminal, broadcast, and network reliability analysis demonstrate that SEN+ and SEN+2 continuously outperform SEN and are very alike in terms of reliability. - Highlights: • The impact of increasing the number of stages on reliability of MINs is investigated. • The RBD method as an accurate method is used for the reliability analysis of MINs. • Complex series–parallel RBDs are used to determine the reliability of the MINs. • All measures of the reliability (i.e. terminal, broadcast, and network reliability) are analyzed. • All reliability equations will be calculated for different size N×N

Improving Stochastic Communication Network Performance: Reliability vs. Throughput

Science.gov (United States)

1991-12-01

increased to one. 2) arc survivabil.. ities will be increased in increments of one tenths. and 3) the costs to increase- arc si’rvivabilities were equal and...This reliability value is leni used to maximize the associated expected flow. For Net work A. a bIdget of (8)() pro(duces a tradcoff point at (.58.37...Network B for a buidgel of 2000 which allows a nel \\\\ork relial)ilitv of one to be achieved and a bidget of 1200 which allows for ;, maximum 57

Highly reliable TOFD UT Technique

International Nuclear Information System (INIS)

Acharya, G.D.; Trivedi, S.A.R.; Pai, K.B.

2003-01-01

The high performance of the time of flight diffraction technique (TOFD) with regard to the detection capabilities of weld defects such as crack, slag, lack of fusion has led to a rapidly increasing acceptance of the technique as a pre?service inspection tool. Since the early 1990s TOFD has been applied to several projects, where it replaced the commonly used radiographic testing. The use of TOM lead to major time savings during new build and replacement projects. At the same time the TOFD technique was used as base line inspection, which enables monitoring in the future for critical welds, but also provides documented evidence for life?time. The TOFD technique as the ability to detect and simultaneously size flows of nearly any orientation within the weld and heat affected zone. TOM is recognized as a reliable, proven technique for detection and sizing of defects and proven to be a time saver, resulting in shorter shutdown periods and construction project times. Thus even in cases where inspection price of TOFD per welds is higher, in the end it will result in significantly lower overall costs and improve quality. This paper deals with reliability, economy, acceptance criteria and field experience. It also covers comparative study between radiography technique Vs. TOFD. (Author)

Stone stability under non-uniform flow

NARCIS (Netherlands)

Hoan, N.T.

2008-01-01

Despite the fact that many studies on the stability of stones in bed protections under flowing water have been conducted, our knowledge is still far from advanced and reliable. Issues like how to quantify the hydraulic loads exerted on the stones on a bed and the associated stability of the stones

Numerical simulation of the flow field in pump intakes by means of Lattice Boltzmann methods

International Nuclear Information System (INIS)

Schneider, A; Conrad, D; Böhle, M

2013-01-01

Lattice Boltzmann Methods are nowadays popular schemes for solving fluid flow problems of engineering interest. This popularity is due to the advantages of these schemes: For example, the meshing of the fluid domain can be performed fully automatically which results in great simplicity in handling complex geometries. In this paper a numerical scheme for the flow simulation in pump intakes based on a Lattice Boltzmann large eddy approach is presented. The ability of this scheme to capture the flow phenomena of the intake flow at different operating conditions is analysed. For the operational reliability and efficiency of pumps and pump systems, the incoming flow conditions are crucial. Since the efficiency and reliability requirements of pumps are rising and must be guaranteed, the flow conditions in pump intakes have to be evaluated during plant planning. Recent trends show that pump intakes are built more and more compact, which makes the flow in the intake even more complex. Numerical methods are a promising technique for conduction flow analysis in pump intakes, because they can be realised rapidly and cheaply

Dynamic surface-pressure instrumentation for rods in parallel flow

International Nuclear Information System (INIS)

Mulcahy, T.M.; Lawrence, W.

1979-01-01

Methods employed and experience gained in measuring random fluid boundary layer pressures on the surface of a small diameter cylindrical rod subject to dense, nonhomogeneous, turbulent, parallel flow in a relatively noise-contaminated flow loop are described. Emphasis is placed on identification of instrumentation problems; description of transducer construction, mounting, and waterproofing; and the pretest calibration required to achieve instrumentation capable of reliable data acquisition

Reliability data banks

International Nuclear Information System (INIS)

Cannon, A.G.; Bendell, A.

1991-01-01

Following an introductory chapter on Reliability, what is it, why it is needed, how it is achieved and measured, the principles of reliability data bases and analysis methodologies are the subject of the next two chapters. Achievements due to the development of data banks are mentioned for different industries in the next chapter, FACTS, a comprehensive information system for industrial safety and reliability data collection in process plants are covered next. CREDO, the Central Reliability Data Organization is described in the next chapter and is indexed separately, as is the chapter on DANTE, the fabrication reliability Data analysis system. Reliability data banks at Electricite de France and IAEA's experience in compiling a generic component reliability data base are also separately indexed. The European reliability data system, ERDS, and the development of a large data bank come next. The last three chapters look at 'Reliability data banks, - friend foe or a waste of time'? and future developments. (UK)

An Introduction To Reliability

International Nuclear Information System (INIS)

Park, Kyoung Su

1993-08-01

This book introduces reliability with definition of reliability, requirement of reliability, system of life cycle and reliability, reliability and failure rate such as summary, reliability characteristic, chance failure, failure rate which changes over time, failure mode, replacement, reliability in engineering design, reliability test over assumption of failure rate, and drawing of reliability data, prediction of system reliability, conservation of system, failure such as summary and failure relay and analysis of system safety.

Designing reliability into high-effectiveness industrial gas turbine regenerators

International Nuclear Information System (INIS)

Valentino, S.J.

1979-01-01

The paper addresses the measures necessary to achieve a reliable regenerator design that can withstand higher temperatures (1000-1200 F) and many start and stop cycles - conditions encountered in high-efficiency operation in pipeline applications. The discussion is limited to three major areas: (1) structural analysis of the heat exchanger core - the part of the regenerator that must withstand the higher temperatures and cyclic duty (2) materials data and material selection and (3) a comprehensive test program to demonstrate the reliability of the regenerator. This program includes life-cycle tests, pressure containment in fin panels, core-to-core joint structural test, bellows pressure containment test, sliding pad test, core gas-side passage flow distribution test, and production test. Today's regenerators must have high cyclic life capability, stainless steel construction, and long fault-free service life of 120,000 hr

S-phase-dependent cell cycle disturbances caused by Aleutian mink disease parvovirus

DEFF Research Database (Denmark)

Oleksiewicz, M.B.; Alexandersen, Søren

1997-01-01

We examined replication of the autonomous parovirus Aleutian mink disease parovirus (ADV) in relation to cell cycle progression of permissive Crandell feline kidney (CRFK) cells. Flow cytometric analysis showed that ADV caused a composite, binary pattern of cell cycle arrest. ADV-induced cell cyc...

Scaling-Laws of Flow Entropy with Topological Metrics of Water Distribution Networks

Directory of Open Access Journals (Sweden)

Giovanni Francesco Santonastaso

2018-01-01

Full Text Available Robustness of water distribution networks is related to their connectivity and topological structure, which also affect their reliability. Flow entropy, based on Shannon’s informational entropy, has been proposed as a measure of network redundancy and adopted as a proxy of reliability in optimal network design procedures. In this paper, the scaling properties of flow entropy of water distribution networks with their size and other topological metrics are studied. To such aim, flow entropy, maximum flow entropy, link density and average path length have been evaluated for a set of 22 networks, both real and synthetic, with different size and topology. The obtained results led to identify suitable scaling laws of flow entropy and maximum flow entropy with water distribution network size, in the form of power–laws. The obtained relationships allow comparing the flow entropy of water distribution networks with different size, and provide an easy tool to define the maximum achievable entropy of a specific water distribution network. An example of application of the obtained relationships to the design of a water distribution network is provided, showing how, with a constrained multi-objective optimization procedure, a tradeoff between network cost and robustness is easily identified.

Reliability calculations

International Nuclear Information System (INIS)

Petersen, K.E.

1986-03-01

Risk and reliability analysis is increasingly being used in evaluations of plant safety and plant reliability. The analysis can be performed either during the design process or during the operation time, with the purpose to improve the safety or the reliability. Due to plant complexity and safety and availability requirements, sophisticated tools, which are flexible and efficient, are needed. Such tools have been developed in the last 20 years and they have to be continuously refined to meet the growing requirements. Two different areas of application were analysed. In structural reliability probabilistic approaches have been introduced in some cases for the calculation of the reliability of structures or components. A new computer program has been developed based upon numerical integration in several variables. In systems reliability Monte Carlo simulation programs are used especially in analysis of very complex systems. In order to increase the applicability of the programs variance reduction techniques can be applied to speed up the calculation process. Variance reduction techniques have been studied and procedures for implementation of importance sampling are suggested. (author)

Reliability estimates for selected sensors in fusion applications

International Nuclear Information System (INIS)

Cadwallader, L.C.

1996-09-01

This report presents the results of a study to define several types of sensors in use, the qualitative reliability (failure modes) and quantitative reliability (average failure rates) for these types of process sensors. Temperature, pressure, flow, and level sensors are discussed for water coolant and for cryogenic coolants. The failure rates that have been found are useful for risk assessment and safety analysis. Repair times and calibration intervals are also given when found in the literature. All of these values can also be useful to plant operators and maintenance personnel. Designers may be able to make use of these data when planning systems. The final chapter in this report discusses failure rates for several types of personnel safety sensors, including ionizing radiation monitors, toxic and combustible gas detectors, humidity sensors, and magnetic field sensors. These data could be useful to industrial hygienists and other safety professionals when designing or auditing for personnel safety

The Combination of Micro Diaphragm Pumps and Flow Sensors for Single Stroke Based Liquid Flow Control

Directory of Open Access Journals (Sweden)

Christoph Jenke

2017-04-01

Full Text Available With the combination of micropumps and flow sensors, highly accurate and secure closed-loop controlled micro dosing systems for liquids are possible. Implementing a single stroke based control mode with piezoelectrically driven micro diaphragm pumps can provide a solution for dosing of volumes down to nanoliters or variable average flow rates in the range of nL/min to μL/min. However, sensor technologies feature a yet undetermined accuracy for measuring highly pulsatile micropump flow. Two miniaturizable in-line sensor types providing electrical readout—differential pressure based flow sensors and thermal calorimetric flow sensors—are evaluated for their suitability of combining them with mircopumps. Single stroke based calibration of the sensors was carried out with a new method, comparing displacement volumes and sensor flow volumes. Limitations of accuracy and performance for single stroke based flow control are described. Results showed that besides particle robustness of sensors, controlling resistive and capacitive damping are key aspects for setting up reproducible and reliable liquid dosing systems. Depending on the required average flow or defined volume, dosing systems with an accuracy of better than 5% for the differential pressure based sensor and better than 6.5% for the thermal calorimeter were achieved.

Is 2D impedance tomography a reliable technique for two-phase flow?

International Nuclear Information System (INIS)

Lemonnier, H.; Peytraud, J.F.

1998-01-01

Impedance tomography consists in reconstructing the conductivity distribution from electrical data which characterize the electrical response of a medium to arbitrary excitations. Impedance tomography is an ill-conditioned problem and designing a tomograph therefore requires the quantitative knowledge of the sensitivity of the reconstruction to the measurements noise. The numerical conditioning of an original and accurate algorithm has been studied. This algorithm does not suffer from the shortcomings already identified in the literature. It is shown that for media encompassing inclusions which is a typical situation in two-phase flows, the necessary accuracy for the measurements if far beyond any technological reach. Moreover, within these high requirements for accuracy, some side effects must be carefully controlled or compensated and relevant procedures arc provided. Furthermore. reconstruction artifacts are shown and they are found to derive from the unavoidable tridimensional nature of the electric field. For all these reasons, it is concluded that impedance tomography has very low potentialities as an accurate phase fraction distribution measuring technique in any arbitrary two-phase flows. (author)

Prediction of Tensile Behavior of UHSFRC Considering the Flow Field in the Placing Dominated by Shear Flow

Directory of Open Access Journals (Sweden)

Joon-Shik Moon

2018-01-01

Full Text Available Considering the case of fabricating a UHSFRC (ultra-high strength fiber-reinforced concrete beam with the method of one end placing and self-flowing to the other end, it was intended to simulate the variation of the fiber orientation distribution according to the flow distance and the variation of the resultant tensile behaviors. Then the validity of the simulation approach was shown by comparing the simulated results with experimental ones. A three-point bending test with a notched beam was adopted for the experiment and a finite element analysis was performed to obtain the simulated results for the bending test considering the flow-dependent tensile behavior of the UHSFRC. From the simulation for the fiber orientation distribution according to the flow distance, it could be found that the major change in the fiber orientation distribution took place within a short flow distance and most of the fibers became nearly aligned to the flow direction. After some flow distance, there was a not-so-remarkable variation in the fiber orientation distribution that could influence the tensile behavior of the composite. For this flow region, the consistent flexural test results, regardless of flow distance, demonstrate the reliability of the simulation.

Report of the results of the International Clinical Cytometry Society and American Society for Clinical Pathology workload survey of clinical flow cytometry laboratories.

Science.gov (United States)

Wolniak, Kristy; Goolsby, Charles; Choi, Sarah; Ali, Asma; Serdy, Nina; Stetler-Stevenson, Maryalice

2017-11-01

Thorough review of current workload, staffing, and testing practices in clinical laboratories allows for optimization of laboratory efficiency and quality. This information is largely missing with regard to clinical flow cytometry laboratories. The purpose of this survey is to provide comprehensive, current, and accurate data on testing practices and laboratory staffing in clinical laboratories performing flow cytometric studies. Survey data was collected from flow cytometry laboratories through the ASCP website. Data was collected on the workload during a 1-year time period of full-time and part-time technical and professional (M.D./D.O./Ph.D. or equivalent) flow cytometry employees. Workload was examined as number of specimens and tubes per full time equivalent (FTE) technical and professional staff. Test complexity, test result interpretation, and reporting practices were also evaluated. There were 205 respondent laboratories affiliated predominantly with academic and health system institutions. Overall, 1,132 FTE employees were reported with 29% professional FTE employees and 71% technical. Fifty-one percent of the testing performed was considered high complexity and 49% was low complexity. The average number of tubes per FTE technologist was 1,194 per year and the average number of specimens per FTE professional was 1,659 per year. The flow cytometry reports were predominantly written by pathologists (57%) and were typically written as a separate report (58%). This survey evaluates the overall status of the current practice of clinical flow cytometry and provides a comprehensive dataset as a framework to help laboratory departments, directors, and managers make appropriate, cost-effective staffing decisions. © 2016 International Clinical Cytometry Society. © 2016 International Clinical Cytometry Society.

Delta flow: An accurate, reliable system for detecting kicks and loss of circulation during drilling

Energy Technology Data Exchange (ETDEWEB)

Speers, J.M.; Gehrig, G.F.

1987-12-01

A system to monitor drilling-fluid flow rate has been developed that detects kicks and lost returns in floating, fixed-platform, and land-base drilling operations. The system uses flowmeters that monitor the flow rates of drilling fluids entering the borehole through the standpipe and leaving the well through the return flowline. These readings are processed in a computer-based, data-acquisition system to form a filtered delta-flow signal that identified the occurrence of downhole fluid gains or losses. The system is designed to trip an alarm when a gain or loss exceeds 25 gal/min (1.6 dm/sup 3//s), even in a floating drilling environment. This sensitivity will generally keep gains or losses to less than 5 bbl (0.8 m/sup 3/).

Reliability assessment of distribution power systems including distributed generations

International Nuclear Information System (INIS)

Megdiche, M.

2004-12-01

Nowadays, power systems have reached a good level of reliability. Nevertheless, considering the modifications induced by the connections of small independent producers to distribution networks, there's a need to assess the reliability of these new systems. Distribution networks present several functional characteristics, highlighted by the qualitative study of the failures, as dispersed loads at several places, variable topology and some electrotechnical phenomena which must be taken into account to model the events that can occur. The adopted reliability calculations method is Monte Carlo simulations, the probabilistic method most powerful and most flexible to model complex operating of the distribution system. We devoted a first part on the case of a 20 kV feeder to which a cogeneration unit is connected. The method was applied to a software of stochastic Petri nets simulations. Then a second part related to the study of a low voltage power system supplied by dispersed generations. Here, the complexity of the events required to code the method in an environment of programming allowing the use of power system calculations (load flow, short-circuit, load shedding, management of units powers) in order to analyse the system state for each new event. (author)

Interactive reliability assessment using an integrated reliability data bank

International Nuclear Information System (INIS)

Allan, R.N.; Whitehead, A.M.

1986-01-01

The logical structure, techniques and practical application of a computer-aided technique based on a microcomputer using floppy disc Random Access Files is described. This interactive computational technique is efficient if the reliability prediction program is coupled directly to a relevant source of data to create an integrated reliability assessment/reliability data bank system. (DG)

Regional cerebral blood flow measurement using a scintillation camera

International Nuclear Information System (INIS)

Heiss, W.D.

1979-01-01

A scintillation camera connected to auxillary equipment with off-line data processing or connected to an on-line dedicated computer system permits measurement of hemispheric and regional cerebral blood flow. Reliable flow values are obtained from regions limited in size by spatial resolution and the count rates achieved. Flow measurements obtained with the camera are able to resolve inhomogeneities of cerebral circulation in normal subjects. In a variety of clinical conditions, the localization, severity and extent of flow alterations are shown. Results of flow measurements in individual cases elucidate the pathogenesis of neurologic deficits, quantify the damage to the brain, indicate therapeutic measures of potential value and permit an estimation of the further clinical course. With restricted spatial resolution, flow measurements after intravenous 133 Xe injection are also feasible

Contactless flowrate sensors for Na, PbBi and Pb flows

International Nuclear Information System (INIS)

Buchenau, D.; Gerbeth, G.; Priede, J.

2011-01-01

Accurate and reliable flow rate measurements are required for various liquid metal systems such as the Na or Lead-flows in fast reactors, the PbBi-flows in transmutation systems, or the flows in liquid metal targets. For liquid metal flows, a contactless measurement is preferable. In this paper we report on the recent development of two types of such flow meters. The former operates by detecting the flow-induced disturbance in the phase distribution of an externally applied AC magnetic field. Such a phase-shift flow meter was developed with an emitting coil at one side of the duct and two sensing coils at the opposite side. The second approach uses a rotatable single cylindrical permanent magnet, which is placed close to the liquid metal duct. The rotation rate of this magnet is proportional to the flow rate. (author)

Climatology of local flow patterns around Basel

Energy Technology Data Exchange (ETDEWEB)

Weber, R.O. [Paul Scherrer Inst. (PSI), Villigen (Switzerland)

1997-06-01

Recently a method has been developed to classify local-scale flow patterns from the wind measurements at a dense network of stations. It was found that in the MISTRAL area around Basel a dozen characteristic flow patterns occur. However, as the dense network of stations ran only during one year, no reliable climatology can be inferred from these data, especially the annual cycle of the flow patterns is not well determined from a single year of observations. As there exist several routinely operated stations in and near the MISTRAL area, a method was searched to identify the local flow patterns from the observations at the few routine stations. A linear discriminant analysis turned out to be the best method. Based of data from 11 stations which were simultaneously operated during 1990-1995 a six-year climatology of the flow patterns could be obtained. (author) 1 fig., 1 tab., 3 refs.

76 FR 23222 - Electric Reliability Organization Interpretation of Transmission Operations Reliability

Science.gov (United States)

2011-04-26

....3d 1342 (DC Cir. 2009). \\5\\ Mandatory Reliability Standards for the Bulk-Power System, Order No. 693... Reliability Standards for the Bulk-Power System. Action: FERC-725A. OMB Control No.: 1902-0244. Respondents...] Electric Reliability Organization Interpretation of Transmission Operations Reliability AGENCY: Federal...

Measurement of stenotic rate and blood flow of carotid artery of the dogs with digital subtraction angiography

International Nuclear Information System (INIS)

Kobayashi, Keisuke; Kagawa, Masaaki; Asai, Masaaki; Yasue, Hiroshi; Kawabata, Kazuhiro; Yue, Shuzengmr

1987-01-01

Hemodynamic analysis of stenotic rate and local mean blood flow of the common carotid artery of the dogs with electromagnetic flowmeter and DSA was evaluated. Measurement of stenotic rate using local profile curve was very accurate and it was though to be useful in evaluation of local blood flow of the cervical carotid artery in the patients with carotid stenosis pre-and postoperatively. Although the measurement of absolute blood flow in the case of known diameter of the vessel is exactly reliable, the measured flow is not so reliable in the clinical application for the difficulty in the accurate measurement of the diameter. But hemodynamic analysis of the relative blood flow in the clinical ground can be estimated from this study. The theory and practical measurement are discussed. (author)

Implicit gas-kinetic unified algorithm based on multi-block docking grid for multi-body reentry flows covering all flow regimes

Science.gov (United States)

Peng, Ao-Ping; Li, Zhi-Hui; Wu, Jun-Lin; Jiang, Xin-Yu

2016-12-01

Based on the previous researches of the Gas-Kinetic Unified Algorithm (GKUA) for flows from highly rarefied free-molecule transition to continuum, a new implicit scheme of cell-centered finite volume method is presented for directly solving the unified Boltzmann model equation covering various flow regimes. In view of the difficulty in generating the single-block grid system with high quality for complex irregular bodies, a multi-block docking grid generation method is designed on the basis of data transmission between blocks, and the data structure is constructed for processing arbitrary connection relations between blocks with high efficiency and reliability. As a result, the gas-kinetic unified algorithm with the implicit scheme and multi-block docking grid has been firstly established and used to solve the reentry flow problems around the multi-bodies covering all flow regimes with the whole range of Knudsen numbers from 10 to 3.7E-6. The implicit and explicit schemes are applied to computing and analyzing the supersonic flows in near-continuum and continuum regimes around a circular cylinder with careful comparison each other. It is shown that the present algorithm and modelling possess much higher computational efficiency and faster converging properties. The flow problems including two and three side-by-side cylinders are simulated from highly rarefied to near-continuum flow regimes, and the present computed results are found in good agreement with the related DSMC simulation and theoretical analysis solutions, which verify the good accuracy and reliability of the present method. It is observed that the spacing of the multi-body is smaller, the cylindrical throat obstruction is greater with the flow field of single-body asymmetrical more obviously and the normal force coefficient bigger. While in the near-continuum transitional flow regime of near-space flying surroundings, the spacing of the multi-body increases to six times of the diameter of the single

Mathematical models for two-phase stratified pipe flow

Energy Technology Data Exchange (ETDEWEB)

Biberg, Dag

2005-06-01

The simultaneous transport of oil, gas and water in a single multiphase flow pipe line has for economical and practical reasons become common practice in the gas and oil fields operated by the oil industry. The optimal design and safe operation of these pipe lines require reliable estimates of liquid inventory, pressure drop and flow regime. Computer simulations of multiphase pipe flow have thus become an important design tool for field developments. Computer simulations yielding on-line monitoring and look ahead predictions are invaluable in day-to-day field management. Inaccurate predictions may have large consequences. The accuracy and reliability of multiphase pipe flow models are thus important issues. Simulating events in large pipelines or pipeline systems is relatively computer intensive. Pipe-lines carrying e.g. gas and liquefied gas (condensate) may cover distances of several hundred km in which transient phenomena may go on for months. The evaluation times associated with contemporary 3-D CFD models are thus not compatible with field applications. Multiphase flow lines are therefore normally simulated using specially dedicated 1-D models. The closure relations of multiphase pipe flow models are mainly based on lab data. The maximum pipe inner diameter, pressure and temperature in a multiphase pipe flow lab is limited to approximately 0.3 m, 90 bar and 60{sup o}C respectively. The corresponding field values are, however, much higher i.e.: 1 m, 1000 bar and 200{sup o}C respectively. Lab data does thus not cover the actual field conditions. Field predictions are consequently frequently based on model extrapolation. Applying field data or establishing more advanced labs will not solve this problem. It is in fact not practically possible to acquire sufficient data to cover all aspects of multiphase pipe flow. The parameter range involved is simply too large. Liquid levels and pressure drop in three-phase flow are e.g. determined by 13 dimensionless parameters

Requirements of safety and reliability

International Nuclear Information System (INIS)

Franzen, L.F.

1977-01-01

The safety strategy for nuclear power plants is characterized by the fact that the high level of safety was attained not as a result of experience, but on the basis of preventive accident analyses and the findings derived from such analyses. Although, in these accident analyses, the deterministic approach is predominant it is supplemented by reliability analyses. The accidents analyzed in nuclear licensing procedures cover a wide spectrum from minor incidents to the design basis accidents which determine the design of the safety devices. The initial and boundary conditions, which are essential for accident analyses, and the determination of the loads occuring in various states during regular operation and in accidents flow into the design of the individual systems and components. The inevitable residual risk and its origins are discussed. (orig./HP) [de

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...

Computer Model to Estimate Reliability Engineering for Air Conditioning Systems

International Nuclear Information System (INIS)

Afrah Al-Bossly, A.; El-Berry, A.; El-Berry, A.

2012-01-01

Reliability engineering is used to predict the performance and optimize design and maintenance of air conditioning systems. Air conditioning systems are expose to a number of failures. The failures of an air conditioner such as turn on, loss of air conditioner cooling capacity, reduced air conditioning output temperatures, loss of cool air supply and loss of air flow entirely can be due to a variety of problems with one or more components of an air conditioner or air conditioning system. Forecasting for system failure rates are very important for maintenance. This paper focused on the reliability of the air conditioning systems. Statistical distributions that were commonly applied in reliability settings: the standard (2 parameter) Weibull and Gamma distributions. After distributions parameters had been estimated, reliability estimations and predictions were used for evaluations. To evaluate good operating condition in a building, the reliability of the air conditioning system that supplies conditioned air to the several The company's departments. This air conditioning system is divided into two, namely the main chilled water system and the ten air handling systems that serves the ten departments. In a chilled-water system the air conditioner cools water down to 40-45 degree F (4-7 degree C). The chilled water is distributed throughout the building in a piping system and connected to air condition cooling units wherever needed. Data analysis has been done with support a computer aided reliability software, this is due to the Weibull and Gamma distributions indicated that the reliability for the systems equal to 86.012% and 77.7% respectively. A comparison between the two important families of distribution functions, namely, the Weibull and Gamma families was studied. It was found that Weibull method performed for decision making.

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.

Inhibition of HIV-1 lentiviral particles infectivity by Gynostemma ...

African Journals Online (AJOL)

These claims motivated the study in which the inhibition of viral vector infectivity of HeLa cells was assessed flow cytometrically by measuring the expression of green fluorescent protein (GFP) transgene incorporated in the lentiviral vector construct. An infectious VSV-G-pseudotyped, human immunodeficiency virus type ...

76 FR 23171 - Electric Reliability Organization Interpretations of Interconnection Reliability Operations and...

Science.gov (United States)

2011-04-26

... Reliability Standards for the Bulk-Power System, Order No. 693, FERC Stats. & Regs. ] 31,242, order on reh'g...-Power System reliability may request an interpretation of a Reliability Standard.\\7\\ The ERO's standards... information in its reliability assessments. The Reliability Coordinator must monitor Bulk Electric System...

Cytometry of mammalian sperm

Energy Technology Data Exchange (ETDEWEB)

Gledhill, B.L.

1983-10-11

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. The accessibility of male cells makes them well suited for analytical cytology. We might automate the process of determining sperm morphology but should not do so solely for increased speed. Rather, richer tangible benefits will derive from cytometric evaluation through increased sensitivity, reduced subjectivity, standardization between investigators and laboratories, enhanced archival systems, and the benefits of easily exchanged standardized data. Inroads on the standardization of assays for motility and functional integrity are being made. Flow cytometric analysis of total DNA content of individual sperm is an insensitive means to detect exposure to reproductive toxins because of the small size and low frequency of the DNA content errors. Flow cytometry can be applied to determine the proportions of X- and Y-sperm in semen samples.

Cytometry of 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. The accessibility of male cells makes them well suited for analytical cytology. We might automate the process of determining sperm morphology but should not do so solely for increased speed. Rather, richer tangible benefits will derive from cytometric evaluation through increased sensitivity, reduced subjectivity, standardization between investigators and laboratories, enhanced archival systems, and the benefits of easily exchanged standardized data. Inroads on the standardization of assays for motility and functional integrity are being made. Flow cytometric analysis of total DNA content of individual sperm is an insensitive means to detect exposure to reproductive toxins because of the small size and low frequency of the DNA content errors. Flow cytometry can be applied to determine the proportions of X- and Y-sperm in semen samples

Measurement of Platelet-derived Microparticle Levels in the Chronic Phase of Cerebral Infarction Using an Enzyme-linked Immunosorbent Assay

OpenAIRE

Shirafuji, Toshihiko; Hamaguchi, Hirotoshi; Kanda, Fumio

2008-01-01

Assessment of platelet function is a critical component of the treatment andsecondary prevention of cerebral infarction, and measurement of platelet-derivedmicroparticle (PDMP) levels using flow cytometry may be a good indicator of plateletfunction. However, the flow cytometric analysis is not feasible in a variety of clinicalsituations. The goal of the present study was to measure PDMP levels using anenzyme-linked immunosorbent assay (ELISA) in chronic cerebral infarction patientsand to dete...

Non-equilibrium effects on the two-phase flow critical phenomenon

International Nuclear Information System (INIS)

Sami, S.M.

1988-01-01

In the present study, the choking criterion for nonhomogeneous nonequilibrium two phase flow is obtained by solving the two-fluid model conservation equations. The method of characteristics is employed to predict the critical flow conditions. Critical flow is established after the magnitude of the characteristic slopes (velocities). Critical flow conditions are reached when the smallest characteristic slope becomes equal to zero. Several expression are developed to determine the nonequilibrium mass and heat exchanges in terms of the system dependent parameters derivatives. In addition, comprehensive transition flow regime maps are employed in the calculation of interfacial heat and momentum transfer rates. Numerical results reveal that the proposed model reliably predicts the critical two-phase flow phenomenon under different inlet conditions and compares well with other existing models

Individual renal plasma flow determination in 2 minutes

International Nuclear Information System (INIS)

Schlegel, J.U.; Hamway, S.A.

1976-01-01

A method is described for determining effective renal plasma flow, using the renal counts 1 to 2 minutes after injection and applying background subtraction as a ratio to the total amount of radioactivity injected intravenously with appropriate correction for kidney depth. It appears that this simple approach for determination of effective renal plasma flow within a matter of 2 minutes is accurate and reliable, and should be useful for a number of applications when it is important to determine total and individual renal plasma flow. The approach is recommended as a screening procedure in lieu of the determination of blood urea nitrogen and serum creatinine, with the additional advantages of gross morphology and determination of individual renal plasma flow in an absolute as well as relative manner

APPLICATION OF TRAVEL TIME RELIABILITY FOR PERFORMANCE ORIENTED OPERATIONAL PLANNING OF EXPRESSWAYS

Science.gov (United States)

Mehran, Babak; Nakamura, Hideki

Evaluation of impacts of congestion improvement scheme s on travel time reliability is very significant for road authorities since travel time reliability repr esents operational performance of expressway segments. In this paper, a methodology is presented to estimate travel tim e reliability prior to implementation of congestion relief schemes based on travel time variation modeling as a function of demand, capacity, weather conditions and road accident s. For subject expressway segmen ts, traffic conditions are modeled over a whole year considering demand and capacity as random variables. Patterns of demand and capacity are generated for each five minute interval by appl ying Monte-Carlo simulation technique, and accidents are randomly generated based on a model that links acci dent rate to traffic conditions. A whole year analysis is performed by comparing de mand and available capacity for each scenario and queue length is estimated through shockwave analysis for each time in terval. Travel times are estimated from refined speed-flow relationships developed for intercity expressways and buffer time index is estimated consequently as a measure of travel time reliability. For validation, estimated reliability indices are compared with measured values from empirical data, and it is shown that the proposed method is suitable for operational evaluation and planning purposes.

AC power flow importance measures considering multi-element failures

International Nuclear Information System (INIS)

Li, Jian; Dueñas-Osorio, Leonardo; Chen, Changkun; Shi, Congling

2017-01-01

Quantifying the criticality of individual components of power systems is essential for overall reliability and management. This paper proposes an AC-based power flow element importance measure, while considering multi-element failures. The measure relies on a proposed AC-based cascading failure model, which captures branch overflow, bus load shedding, and branch failures, via AC power flow and optimal power flow analyses. Taking the IEEE 30, 57 and 118-bus power systems as case studies, we find that N-3 analyses are sufficient to measure the importance of a bus or branch. It is observed that for a substation bus, its importance is statistically proportional to its power demand, but this trend is not observed for power plant buses. While comparing with other reliability, functionality, and topology-based importance measures popular today, we find that a DC power flow model, although better correlated with the benchmark AC model as a whole, still fails to locate some critical elements. This is due to the focus of DC-based models on real power that ignores reactive power. The proposed importance measure is aimed to inform decision makers about key components in complex systems, while improving cascading failure prevention, system backup setting, and overall resilience. - Highlights: • We propose a novel importance measure based on joint failures and AC power flow. • A cascading failure model considers both AC power flow and optimal power flow. • We find that N-3 analyses are sufficient to measure the importance of an element. • Power demand impacts the importance of substations but less so that of generators. • DC models fail to identify some key elements, despite correlating with AC models.

Measurement of blowdown flow rates using load cells

International Nuclear Information System (INIS)

Dolas, P.K.; Venkat Raj, V.; Ghosh, A.K.; Murty, L.G.K.; Muralidhar Rao, S.

1980-01-01

To establish a reliable method for measuring two-phase flow, experiments were planned for measurement of transient single phase flow rates from vessels using load cells. Suitability of lead-zirconate-titanate piezoelectric ceramic discs was examined. Discharge time constant of the disc used was low, leading to large measurement errors. Subsequently, experiments were carried out using strain gauge load cells and these were found satisfactory. The unsteady flow equation has been derived for the system under investigation. The equation has been solved numerically using the fourth order Runge-Kutta method and also by integrating it analytically. The experimental results are compared with the theoretical results and presented in this report. (auth.)

Postfact phenomena of the wet-steam flow electrization in turbines

Science.gov (United States)

Tarelin, A. A.

2017-11-01

Physical processes occurring in a turbine with natural electrization of a humidity-steam flow and their effect on efficiency and reliability of the turbine operation has been considered. Causes of the electrical potential occurrence on a rotor shaft are analyzed. The wet steam's electrization exposure on the electrical potential that is one of the major factors of bearings' electroerosion has been demonstrated on the full-scale installation. Hydrogen formation in wheelspace of the turbine as a result of electrochemical processes and electric field exposure of the space charge has been considered. Hydrogen concentration dependence on a volume charge density in the steam flow has been determined. It is stated that the processes occurring behind the final stage of wet-steam turbines are similar to the ones in elaerosol ectrostatic generators. It has been demonstrated that this phenomenon causes the flow's temporal inhibition and starts pulsations. These factors' impact on power loss of the turbine has been evaluated and recommendations for their elimination have been offered. It has been determined that motions of charged drops can cause self-maintained discharges inside of the flow and between the flow and grounded surfaces that are accompanied by electromagnetic radiation of the wide spectrum. The integrated studies have shown that physical phenomena occurring due to natural electrization negatively affect efficiency and reliability of the turbine operation. Practical recommendations allowing one to minimize the negative effects of the flow natural electrization process have been offered.

Handheld Fluorescence Microscopy based Flow Analyzer.