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Sample records for cytometric micronucleus scoring

  1. 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.

  2. Micronucleus test for radiation biodosimetry in mass casualty events: Evaluation of visual and automated scoring

    Energy Technology Data Exchange (ETDEWEB)

    Bolognesi, Claudia, E-mail: claudia.bolognesi@istge.i [Environmental Carcinogenesis Unit, National Cancer Research Institute, Largo R. Benzi 10, 16132 Genoa (Italy); Balia, Cristina; Roggieri, Paola [Environmental Carcinogenesis Unit, National Cancer Research Institute, Largo R. Benzi 10, 16132 Genoa (Italy); Cardinale, Francesco [Clinical Epidemiology Unit, National Cancer Research Institute, Largo R. Benzi 10, 16132 Genoa (Italy); Department of Health Sciences, University of Genoa, Genoa (Italy); Bruzzi, Paolo [Clinical Epidemiology Unit, National Cancer Research Institute, Largo R. Benzi 10, 16132 Genoa (Italy); Sorcinelli, Francesca [Environmental Carcinogenesis Unit, National Cancer Research Institute, Largo R. Benzi 10, 16132 Genoa (Italy); Laboratory of Genetics, Histology and Molecular Biology Section, Army Medical and Veterinary, Research Center, Via Santo Stefano Rotondo 4, 00184 Roma (Italy); Lista, Florigio [Laboratory of Genetics, Histology and Molecular Biology Section, Army Medical and Veterinary, Research Center, Via Santo Stefano Rotondo 4, 00184 Roma (Italy); D' Amelio, Raffaele [Sapienza, Universita di Roma II Facolta di Medicina e Chirurgia and Ministero della Difesa, Direzione Generale Sanita Militare (Italy); Righi, Enzo [Frascati National Laboratories, National Institute of Nuclear Physics, Via Enrico Fermi 40, 00044 Frascati, Rome (Italy)

    2011-02-15

    In the case of a large-scale nuclear or radiological incidents a reliable estimate of dose is an essential tool for providing timely assessment of radiation exposure and for making life-saving medical decisions. Cytogenetics is considered as the 'gold standard' for biodosimetry. The dicentric analysis (DA) represents the most specific cytogenetic bioassay. The micronucleus test (MN) applied in interphase in peripheral lymphocytes is an alternative and simpler approach. A dose-effect calibration curve for the MN frequency in peripheral lymphocytes from 27 adult donors was established after in vitro irradiation at a dose range 0.15-8 Gy of {sup 137}Cs gamma rays (dose rate 6 Gy min{sup -1}). Dose prediction by visual scoring in a dose-blinded study (0.15-4.0 Gy) revealed a high level of accuracy (R = 0.89). The scoring of MN is time consuming and requires adequate skills and expertise. Automated image analysis is a feasible approach allowing to reduce the time and to increase the accuracy of the dose estimation decreasing the variability due to subjective evaluation. A good correlation (R = 0.705) between visual and automated scoring with visual correction was observed over the dose range 0-2 Gy. Almost perfect discrimination power for exposure to 1-2 Gy, and a satisfactory power for 0.6 Gy were detected. This threshold level can be considered sufficient for identification of sub lethally exposed individuals by automated CBMN assay.

  3. Automatic analysis of the micronucleus test in primary human lymphocytes using image analysis.

    Science.gov (United States)

    Frieauff, W; Martus, H J; Suter, W; Elhajouji, A

    2013-01-01

    The in vitro micronucleus test (MNT) is a well-established test for early screening of new chemical entities in industrial toxicology. For assessing the clastogenic or aneugenic potential of a test compound, micronucleus induction in cells has been shown repeatedly to be a sensitive and a specific parameter. Various automated systems to replace the tedious and time-consuming visual slide analysis procedure as well as flow cytometric approaches have been discussed. The ROBIAS (Robotic Image Analysis System) for both automatic cytotoxicity assessment and micronucleus detection in human lymphocytes was developed at Novartis where the assay has been used to validate positive results obtained in the MNT in TK6 cells, which serves as the primary screening system for genotoxicity profiling in early drug development. In addition, the in vitro MNT has become an accepted alternative to support clinical studies and will be used for regulatory purposes as well. The comparison of visual with automatic analysis results showed a high degree of concordance for 25 independent experiments conducted for the profiling of 12 compounds. For concentration series of cyclophosphamide and carbendazim, a very good correlation between automatic and visual analysis by two examiners could be established, both for the relative division index used as cytotoxicity parameter, as well as for micronuclei scoring in mono- and binucleated cells. Generally, false-positive micronucleus decisions could be controlled by fast and simple relocation of the automatically detected patterns. The possibility to analyse 24 slides within 65h by automatic analysis over the weekend and the high reproducibility of the results make automatic image processing a powerful tool for the micronucleus analysis in primary human lymphocytes. The automated slide analysis for the MNT in human lymphocytes complements the portfolio of image analysis applications on ROBIAS which is supporting various assays at Novartis.

  4. Duodenal crypt health following exposure to Cr(VI): Micronucleus scoring, γ-H2AX immunostaining, and synchrotron X-ray fluorescence microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Thompson, Chad M.; Wolf, Jeffrey C.; Elbekai, Reem H.; Paranjpe, Madhav G.; Seiter, Jennifer M.; Chappell, Mark A.; Tappero, Ryan V.; Suh, Mina; Proctor, Deborah M.; Bichteler, Anne; Haws, Laurie C.; Harris, Mark A.

    2015-08-01

    Lifetime exposure to high concentrations of hexavalent chromium [Cr(VI)] in drinking water results in intestinal damage and an increase in duodenal tumors in B6C3F1 mice. To assess whether these tumors could be the result of a direct mutagenic or genotoxic mode of action, we conducted a GLP-compliant 7-day drinking water study to assess crypt health along the entire length of the duodenum. Mice were exposed to water (vehicle control), 1.4, 21, or 180 ppm Cr(VI) via drinking water for 7 consecutive days. Crypt enterocytes in Swiss roll sections were scored as normal, mitotic, apoptotic, karyorrhectic, or as having micronuclei. A single oral gavage of 50 mg/kg cyclophosphamide served as a positive control for micronucleus induction. Exposure to 21 and 180 ppm Cr(VI) significantly increased the number of crypt enterocytes. Micronuclei and γ-H2AX immunostaining were not elevated in the crypts of Cr(VI)-treated mice. In contrast, treatment with cyclophosphamide significantly increased numbers of crypt micronuclei and qualitatively increased γ-H2AX immunostaining. Synchrotron-based X-ray fluorescence (XRF) microscopy revealed the presence of strong Cr fluorescence in duodenal villi, but negligible Cr fluorescence in the crypt compartment. Together, these data indicate that Cr(VI) does not adversely effect the crypt compartment where intestinal stem cells reside, and provide additional evidence that the mode of action for Cr(VI)-induced intestinal cancer in B6C3F1 mice involves chronic villous wounding resulting in compensatory crypt enterocyte hyperplasia.

  5. 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

  6. Micronucleus assay for radiation workers

    International Nuclear Information System (INIS)

    Balasem, A.N.; Ali, A.S.K.

    1997-01-01

    Micronucleus assay was performed on 49 radiation workers and 22 healthy volunteers. Radiation workers were subdivided into two groups according to their employments durations in the radiation field. Group a consisted of 18 radiation workers who have been in this work between 5 and 22 years. Group b included 31 employees who have been classified as radiation workers for 1 to 4.5 years. Statistical analysis showed significant variations between the yields of micronuclei in groups A and B as well as between group A and a group of healthy controls. Meanwhile no significant difference was noticed between the yields of micronuclei in group B and the corresponding values in the healthy controls. The possible effect of age in the induction of micronuclei was discussed and a comparison with the yield of chromosomal aberrations was described. It seems that cytokinesis- blocking method may be used to detect the radiation-induced micronuclei in workers exposed occupationally to ionizing radiation in levels below the maximum permissible limit of 0.05 Sv per year

  7. CYTOKINESIS-BLOCK MICRONUCLEUS ASSAY IN HUMAN GLIOMA CELLS EXPOSED TO RADIATION

    Directory of Open Access Journals (Sweden)

    Jerzy Slowinski

    2011-05-01

    Full Text Available Biological tests are efficient in reflecting the biological influences of several types of generally harmful exposures. The micronucleus assay is widely used in genotoxicity studies or studies on genomic damage in general. We present methodological aspects of cytokinesis-block micronucleus assay performed in human gliomas irradiated in vitro. Eight human glioblastoma cell lines obtained from DSMZ (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Germany were gamma-irradiated (60Co over a dose range of 0-10 Gy. Cytokinesis-block micronucleus assay was performed to quantitate cytogenetic damage. The cells were fixed directly on dishes, stained with fluorochrome DAPI and evaluated under fluorescent and phase contrast microscope. The micronucleus frequency was expressed as a micronuclei (MN per binucleated cell (BNC ratio, calculated after scoring at least 100 BNC per dish. The frequency of spontaneous MN ranged from 0.17 to 0.613 (mean: 0.29 ± 0.14. After irradiation increase of MN frequency in the range of 0.312 - 2.241 (mean: 0.98 ± 0.68 was found at 10 Gy. Gliomas are extremely heterogenous in regard to cytogenetic effects of irradiation, as shown in this study by cytokinesis-block micronucleus assay. This test is easily performed on irradiated glioma cell lines and can assist in determining their radiosensitivity. However, in order to obtain reliable and reproducible results, precise criteria for MN scoring must be strictly followed. Simultaneous use of fluorescent and phase contrast equipment improves imaging of morphological details and can further optimize MN scoring.

  8. Micronucleus induction by repeated exposure of diagnostic X-ray on oral buccal mucosa

    International Nuclear Information System (INIS)

    Lohith Tejashvi, K.; Suchetha Kumari, N.; Shetty, Shishir Ram

    2012-01-01

    Radiography is the important diagnostic tools essential for diagnosis and planning of orthodontic treatment. X-ray is ionizing radiation which showed various effects include breaking the bond of biological molecules, inducing loss of ability of cell death, increases nuclear alterations. Micronuclei - x000D - (MN) are small chromatin bodies that appear in the cytoplasm by the - x000D - condensation of acrocentric chromosomal fragments or by whole chromosomes. This - x000D - is a sensitive indicator of genetic damage. - x000D - x000D - . To evaluate micronucleus induction by repeated exposure of diagnostic X-ray on human buccal cell. Methods: 25 patients who visiting to ABSMIDS, Department of Oral medicine and Radiology for dental checkup exposed to diagnostic X-ray more than 4 times have been selected for this study. The buccal cell for analysis was collected from the cheek mucosa by means of gentle scraping of epithelial using ice-cream sticks and placed in Buffer saline. This sample was smeared on glass slide and then fixed in methanol:glacial acetic acid (3:1). Air dried and stained with Giemsa for 15-25 minutes. Then 250 cells in each slides were analyzed under microscope and frequency of micronucleus was scored (n=4). Repeated X-ray exposed cells showed micronucleus (1.25%) and nuclear alteration (2.3%) compare to the control. Repeated X-ray exposure leads to induces detectable number of micronucleus and nuclear alterations. (author)

  9. 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

  10. 40 CFR 79.64 - In vivo micronucleus assay.

    Science.gov (United States)

    2010-07-01

    ... micronucleus assay. (a) Purpose. The micronucleus assay is an in vivo cytogenetic test which uses erythrocytes... that, because it contains RNA, can be differentiated by appropriate staining techniques from a normochromatic erythrocyte (NCE), which lacks RNA. In one to two days, a PCE matures into a NCE. (c) Test method...

  11. Flow cytometric characterization of cerebrospinal fluid cells.

    Science.gov (United States)

    de Graaf, Marieke T; de Jongste, Arjen H C; Kraan, Jaco; Boonstra, Joke G; Sillevis Smitt, Peter A E; Gratama, Jan W

    2011-09-01

    Flow cytometry facilitates the detection of a large spectrum of cellular characteristics on a per cell basis, determination of absolute cell numbers and detection of rare events with high sensitivity and specificity. White blood cell (WBC) counts in cerebrospinal fluid (CSF) are important for the diagnosis of many neurological disorders. WBC counting and differential can be performed by microscopy, hematology analyzers, or flow cytometry. Flow cytometry of CSF is increasingly being considered as the method of choice in patients suspected of leptomeningeal localization of hematological malignancies. Additionally, in several neuroinflammatory diseases such as multiple sclerosis and paraneoplastic neurological syndromes, flow cytometry is commonly performed to obtain insight into the immunopathogenesis of these diseases. Technically, the low cellularity of CSF samples, combined with the rapidly declining WBC viability, makes CSF flow cytometry challenging. Comparison of flow cytometry with microscopic and molecular techniques shows that each technique has its own advantages and is ideally combined. We expect that increasing the number of flow cytometric parameters that can be simultaneously studied within one sample, will further refine the information on CSF cell subsets in low-cellular CSF samples and enable to define cell populations more accurately. Copyright © 2011 International Clinical Cytometry Society.

  12. Micronucleus frequency and content in healthy relatives of cancer patients

    Czech Academy of Sciences Publication Activity Database

    Zölzer, F.; Křížová, M.; Skalická, Z.F.; Rössnerová, Andrea; Šrám, Radim

    2017-01-01

    Roč. 22, č. 7 (2017), s. 667-673 ISSN 1354-750X Institutional support: RVO:68378041 Keywords : micronucleus centromere test * chromosomal damage * inheritable trait Subject RIV: EB - Genetics ; Molecular Biology OBOR OECD: Human genetics Impact factor: 2.006, year: 2016

  13. What Can a Micronucleus Teach? Learning about Environmental Mutagenesis

    Science.gov (United States)

    Linde, Ana R.; Garcia-Vazquez, Eva

    2009-01-01

    The micronucleus test is widely employed in environmental health research. It can also be an excellent tool for learning important concepts in environmental health. In this article we present an inquiry-based laboratory exercise where students explore several theoretical and practical aspects of environmental mutagenesis employing the micronucleus…

  14. Micronucleus induction as a measure of I-131 exposure

    Energy Technology Data Exchange (ETDEWEB)

    Kasuba, V; Horvat, D [Inst. for Medical Recearch and Occupational Health, Zagreb (Croatia). Laboratory for Mutagenesis; Kusic, Z [Clinical Hospital Sestre Milosrdnice, Zagreb (Croatia). Dept. of Oncology and Nuclear Medicine; Vlatkovic, M [Clinical Hospital Centre, Zagreb (Croatia). Dept. of Nuclear Medicine and Radiation Protection

    1994-10-01

    The change of cell numbers in the peripheral blood following irradiation has been studied for many years, particularly in patients undergoing radiotherapy. Recently, attention is directed towards the use of cytogenetic-mutagenetic methods to estimate the biological effects of received radiation dose. The aim of our study was to identify the difference in number and distribution of micronucleus, depending of applied therapeutic dose of iodine-131. According to their diagnosis, six patients have received iodine-131 in range from 80 to 140 mCi, while in the other group of patients the dose values varied from 7 to 32 mCi. On in vitro peripheral blood lymphocyte cultures micronucleus test was applied. Micronucleus analyses were carried out before the treatment, 24, 48 and 96 hours after the oral application of radiopharmaceutical. The number of micronucleus is showing increase, depending on applied radioactivity of iodine-131 and duration of exposition. The clear dose response relationship was never found. These results illustrate the problem associated with the inhomogeneous distribution of dose which results from the concentration of incorporated radionuclide into thyroid or other tissues. (author).

  15. The role of the micronucleus in stomatogenesis in sexual reproduction of Paramecium tetraurelia: laser microbeam irradiation of the micronucleus

    International Nuclear Information System (INIS)

    Tam Laiwa; Ng, S.F.

    1986-01-01

    Fifteen amicronucleate cell lines and 22 cell lines with defective micronuclei were obtained following selective laser microbeam irradiation of the micronucleus. The amicronucleate cell lines showed reduced growth rate and formed abnormal oral apparatuses in asexual reproduction, and failed to produce any oral apparatus in autogamy. The 22 cell lines with defective micronucleus exhibited various abnormalities of the oral apparatus newly formed during autogamy. These abnormalities included the arrest of membranelle assembly, reduction in the length of the buccal cavity and oral membranelles, disruption of the organization of the membranelles, quadrulation of the dorsal peniculus, and failure of addition of membranellar basal body rows. Hence the micronucleus plays multiple roles in sexual stomatogenesis. Our results agree with the notion that the micronucleus acts during a critical period between the second meiotic division and up to the formation of the zygotic nucleus to control the early stage of oral membranelle assembly. Laser microbeam irradiation might have created recessive mutations and/or chromosomal aberrations, which were expressed during this critical period with the formation of abnormal postmeiotic nuclei. (author)

  16. Role of the micronucleus in stomatogenesis in sexual reproduction of Paramecium tetraurelia: laser microbeam irradiation of the micronucleus

    Energy Technology Data Exchange (ETDEWEB)

    Tam Laiwa; Ng, S.F.

    1986-12-01

    Fifteen amicronucleate cell lines and 22 cell lines with defective micronuclei were obtained following selective laser microbeam irradiation of the micronucleus. The amicronucleate cell lines showed reduced growth rate and formed abnormal oral apparatuses in asexual reproduction, and failed to produce any oral apparatus in autogamy. The 22 cell lines with defective micronucleus exhibited various abnormalities of the oral apparatus newly formed during autogamy. These abnormalities included the arrest of membranelle assembly, reduction in the length of the buccal cavity and oral membranelles, disruption of the organization of the membranelles, quadrulation of the dorsal peniculus, and failure of addition of membranellar basal body rows. Hence the micronucleus plays multiple roles in sexual stomatogenesis. Our results agree with the notion that the micronucleus acts during a critical period between the second meiotic division and up to the formation of the zygotic nucleus to control the early stage of oral membranelle assembly. Laser microbeam irradiation might have created recessive mutations and/or chromosomal aberrations, which were expressed during this critical period with the formation of abnormal postmeiotic nuclei.

  17. Micronucleus formation in cultured human keratinocytes: Involvement of intercellular bioactivation.

    Science.gov (United States)

    van Pelt, F N; Haring, R M; Weterings, P J

    1991-01-01

    Micronucleus formation in cultured human keratinocytes was studied after exposure to benzo[a]pyrene, cyclophosphamide and 12-O-tetradecanoylphorbol-13-acetate without the addition of an exogenous metabolizing system. The first two agents need bioactivation by specific isoenzymes of cytochrome P-450 to form genotoxic intermediates. Benzo[a]pyrene induced the micronucleus formation in both uninduced and Aroclor 1254-pretreated cultures. Clastogenic effects of cyclophosphamide were observed only in Aroclor 1254-pretreated cells. The tumour promotor 12-O-tetradecanoylphorbol-13-acetate did not affect the frequency of micronuclei in human keratinocytes. The data indicate that cultured human keratinocytes can be used to study the tissue-specific response to genotoxic agents as well as interindividual variation in biotransformation capacity.

  18. Micronucleus formation in cultured human keratinocytes following exposure to mitomycin C and cyclophosphamide.

    Science.gov (United States)

    van Pelt, F N; Haring, R M; Overkamp, M J; Weterings, P J

    1991-02-01

    A method is described to investigate the induction of micronuclei in cultured human keratinocytes after short-term exposure to known clastogenic agents. The cytokinesis-block method was applied to facilitate the scoring of micronucleated cells. Mitomycin C, a direct-acting compound, caused a 5-20-fold increase in micronuclei over the controls at the highest concentration tested (1 microgram/ml). Cyclophosphamide, an agent requiring metabolic activation, did not induce the formation of micronuclei in cultured keratinocytes. However, after pretreatment of the keratinocyte cultures with Aroclor 1254 for 72 h, exposure to cyclophosphamide resulted in a 3-fold increase in micronucleus frequency over the controls. No cytogenetic effect of Aroclor 1254 was observed in control experiments.

  19. Cytokinesis-block micronucleus method in micro-blood cultures

    International Nuclear Information System (INIS)

    Liu Jinwen; Wang Lianzhi; Yang Cangzhen; Yao Yanyu

    1991-01-01

    This paper reports the cytokinesis-block micronucleus method in micro-blood cultures. The observations on detection induced micronuclei of different doses of 60 Co γ-rays irradiation and spontaneous micronucleus of different ages were performed with CB method in comporison with conventional micronucleus (CM) method. The results showed that with direct peripheral micro-blood cultures the cytoknesis-block micronuclei is also obtained. Using CB method, the micronuclei fequency of different ages was linear relationship, Y = 1.62 + 0.74 D, the spontaneous micronuclei frequency of different ages was 4.14%, the induced micronuclei also was a linear relationship, Y = 6.01 + 0.692 D. Using CM method, it showed that the induced micronuclei was a linear relationship, Y = 0.486 D - 1.968, but there is no significant difference between the micronuclei frequency of different ages. Comparison with CM and direct blood smear methods confirmed that the cytokinesis-block method of micro-blood cultures is more sensitive and precise

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

    African Journals Online (AJOL)

    Jane

    2011-10-24

    Oct 24, 2011 ... daughter cells, leading to a characteristic flow cytometric profile where a ... cell recognition without any impact on bone marrow hemato- ... cells of various cancer cells that load CFSE concentration ... (B) Target cells (R1) were further analyzed in an FL1/FL3 dot plot, ..... hematopoietic cell transplantation.

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

    African Journals Online (AJOL)

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

  2. Cytogenetic Monitoring By Use Of The Micronucleus Assay Among Nuclear Malaysia Radiation Workers-A Preliminary Result

    International Nuclear Information System (INIS)

    Rahimah Abdul Rahim; Mohd Rodzi Ali; Noraisyah Mohd Yusof; Juliana Mahamad Napiah; Yahaya Talib; Rehir Dahalan

    2014-01-01

    Biological dosimetry based on the analysis of micronuclei in the cytokinesis-block micronucleus (CBMN) assay can be used as an alternative method for scoring dicentric chromosomes in the field of radiation protection. Bio dosimetry is mainly performed, in addition to physical dosimetry, with the aim of individual dose assessment. Aim of this study was to assess occupationally induced chromosomal damage in radiation workers exposed to ionizing radiation. The CBMN assay was used in the peripheral blood lymphocytes of 50 exposed workers. Number of bi-nucleated cell and micronuclei were scored and statistical analysis was done to see the effect and correlation of micronuclei with gender, age and time of worked. In conclusion, scoring of micronuclei is a useful cytogenetic monitoring for radiation workers. (author)

  3. Dose response curve for micronucleus of cytokinesis-block method in human lymphocytes after 60Co-gamma ray exposure

    International Nuclear Information System (INIS)

    Gao Jinsheng; Zheng Siying; Cai Feng

    1993-08-01

    The micronucleus technique of cytokines block has been proposed as a new method to measure chromosome damage in cytogenetic. The cytokines is blocked by using cytochalasin B (Cyt-B), and micronuclei are scored in cytokines-blocked (CB) cells. This can easily be done owing to the appearance of binucleate cells and large numbers accumulated by adding 3.0 μg/ml cytochalasin B at 44 hours and scoring at 72 hours. The results show that the optimum concentration of Cyt-B is 3.0 μg/ml. the Cyt-B itself can not induce the increase of micronuclei. The micronucleus frequency of normal individuals in vivo, there is an approximately linear relationship between the frequency of induced micronuclei and irradiation dose. The formula is Y 0.36 D + 2.74 (γ 2 = 0.995 P<0.01). Because the cytokines block method is simple and reliable, it is effective for assaying chromosome damage caused by genetic toxic materials

  4. Low dose effects detected by micronucleus assay in lymphocytes

    International Nuclear Information System (INIS)

    Koeteles, G.J.; Bojtor, I.; Kubasova, T.; Horvath, G.

    1997-01-01

    The effects of low doses of X-rays between 0.01 and 1 Gy were studied on whole blood samples of various individuals using the cytokinesis-blocked lymphocyte micronucleus assay as an endpoint. The adaptive response could be induced in G 0 cells by 0.01 Gy followed by 1 Gy challenging dose within a time period of 8 hours, in vitro. The probability distribution of micronucleus increments in those samples which had received very low doses in the range 0.01-0.05 Gy proved to be of asymmetrical type (i.e. lognormal) -very likely to the same shape which has been verified for unirradiated (control) population - while the variable turned to be normally distributed at or above 1 Gy. Profound changes have been experienced in the main characteristics of the linear dose - response relationship and in regression parameters, as well, when successively lessened dose ranges were studied toward 0.01 Gy. In the range below ∼ 0.2 Gy the response were found to be unrelated to the absorbed dose. These findings suggest that in (very) low dose range a higher attention should be needed to biological parameters like repair, protective mechanisms and antioxidant capacities, rather than to the absorbed radiation energy only. (author)

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

    Science.gov (United States)

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

    2018-02-01

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

  6. Flow cytometry based micronucleus assay for evaluation of genotoxic potential of 2-ACBs in hepatic cells HepG2

    International Nuclear Information System (INIS)

    Barbezan, Angélica B.; Santos, Carla J.B.; Carvalho, Luma R.; Vieira, Daniel P.; Villavicêncio, Anna L.C.H.; Santelli, Glaucia M.M.

    2017-01-01

    Food irradiation is approved for use in more than 60 countries for applications and purposes in a wide variety of foods, being an effective and safe method for preservation and long-term storage. 2-Alkylcyclobutanones (2-ACBs) are the only known radiolytic products generated from foods that contain fatty acids (Triglycerides) when irradiated. The acids analyzed in this study are palmitic and stearic, which when irradiated form 2-Dodecylcyclobutanones (2-dDCB) and 2-Tetradecylcyclobutanone (2-tDCB). Part of the 2-ACBs ingested is excreted through feces and part is deposited in adipose tissues. In vitro studies so far have been only in colon cells. The work used a human hepatoma cell line (HepG2) since the accumulation of fat in this organ is quite common. Micronucleus test was selected to evaluate possible genotoxic effects of 2-dDCB and 2-tDCB compounds when exposed to high concentrations (447, 1422 and 2235 μM) for 4 and 24 hours. Tests were performed in quadriplicates using flow cytometric analysis. None detectable genotoxic damage was observed after 4 hours of exposure to the compounds, and cytotoxic effects were only significant at the highest concentration (2235 μM) of 2-dDCB. After 24 hours of exposure, slight genotoxic damage was observed at all concentrations evaluated, and cytotoxic effects were only present when exposed to compound 2-tDCB. Although there is a genotoxic and cytotoxic effect in some of the situations tested, the two compounds predominantly induced proliferation reduction effects of this hepatic tumor cell line. (author)

  7. Flow cytometry based micronucleus assay for evaluation of genotoxic potential of 2-ACBs in hepatic cells HepG2

    Energy Technology Data Exchange (ETDEWEB)

    Barbezan, Angélica B.; Santos, Carla J.B.; Carvalho, Luma R.; Vieira, Daniel P.; Villavicêncio, Anna L.C.H., E-mail: abarbezan@ipen.br [Instituto de Pesquisas Energéticas e Nucleares (IPEN/CNEN-SP), São Paulo, SP (Brazil); Santelli, Glaucia M.M. [Universidade de São Paulo (USP), SP (Brazil). Departamento de Biologia Celular e do Desenvolvimento

    2017-07-01

    Food irradiation is approved for use in more than 60 countries for applications and purposes in a wide variety of foods, being an effective and safe method for preservation and long-term storage. 2-Alkylcyclobutanones (2-ACBs) are the only known radiolytic products generated from foods that contain fatty acids (Triglycerides) when irradiated. The acids analyzed in this study are palmitic and stearic, which when irradiated form 2-Dodecylcyclobutanones (2-dDCB) and 2-Tetradecylcyclobutanone (2-tDCB). Part of the 2-ACBs ingested is excreted through feces and part is deposited in adipose tissues. In vitro studies so far have been only in colon cells. The work used a human hepatoma cell line (HepG2) since the accumulation of fat in this organ is quite common. Micronucleus test was selected to evaluate possible genotoxic effects of 2-dDCB and 2-tDCB compounds when exposed to high concentrations (447, 1422 and 2235 μM) for 4 and 24 hours. Tests were performed in quadriplicates using flow cytometric analysis. None detectable genotoxic damage was observed after 4 hours of exposure to the compounds, and cytotoxic effects were only significant at the highest concentration (2235 μM) of 2-dDCB. After 24 hours of exposure, slight genotoxic damage was observed at all concentrations evaluated, and cytotoxic effects were only present when exposed to compound 2-tDCB. Although there is a genotoxic and cytotoxic effect in some of the situations tested, the two compounds predominantly induced proliferation reduction effects of this hepatic tumor cell line. (author)

  8. Micronucleus frequency in exfoliated buccal cells from hairdresser who expose to hair products

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    Koh Hui Yee

    2015-06-01

    Full Text Available Background: Hairdresser is one of the fastest growing occupations in today’s society. Hairdresser help styling, cutting, colouring, perming, curling, straightening hair and various treatment to customer. Somehow, hairdresser are constantly exposed to chemical substances such as aromatic amines, hydrogen peroxide, thioglycolic acid, formaldehyde in hair products which can cause damage to human’s genome. Micronucleus is one of the effective biomarker for processes associated with the induction of DNA damage. Purpose: The aim of this study was to determine the micronucleus frequencies in buccal mucosa epithelial cells of hairdresser who were exposed to chemical of hair products. Method: This study was conducted on twenty female subjects, who were divided into 2 groups: exposed and non-exposed (control group. All subjects recruited were working in the same beauty salon. Buccal cells were obtained from each individual by using cytobrush. The cells were stained with modified Feulgen-Ronssenback method and counting of micronucleus per 1000 cell was done under light microscope. The data were analyzed using independent t-test and one-way Anova (p<0.05. Result: The result showed a significant difference in micronucleus frequency between 2 groups. There were a significantly increase of micronucleus frequency in hairdressers and increase of  micronucleus frequency with the longer duration of exposure. Conclusion: It concluded that the chemical substances of hair products had affected the micronucleus frequency ofthe epithelial cells in buccal mucosa of hairdressers.

  9. Cytological assay of micronucleus induction by radiation in oral cancer

    International Nuclear Information System (INIS)

    Bhattathiri, V.N.; Bindu, L.; Remani, P.; Chandralekha, B.; Davis, C.A.; Krishnan Nair, M.

    1996-01-01

    A study was conducted to analyze the dose-response relationship of micronucleus (MN) induction by radiation in eighty-three patients with oral cancers. Serial scrape smears were taken before treatment and after delivery of various fractions of a course of radical radiotherapy. The smears were stained with Giemsa's stain and frequency of micronucleated cells (MNC) evaluated. Before treatment 70.5% of tumours showed MNC, the mean value being 4.18 MNC/1000 cells. The frequency of MNC, increased with increasing dose of radiation. As regards relation to treatment duration, there was initially a slight increase, followed by rapid increase and then a plateauing. Radiosensitive and resistant tumours showed differing pattern of change. The MN test by serial cytological assay has potential as a tool to understand the dynamics of radiation induced cell death and predict radiosensitivity. (author). 4 refs., 5 figs

  10. An improved in vitro micronucleus assay to biological dosimetry

    International Nuclear Information System (INIS)

    Ocampo, Ivette Z.; Okazaki, Kayo; Vieira, Daniel P.

    2013-01-01

    The biological dosimetry is widely used to estimate the absorbed dose in people occupationally or accidentally exposed to the radiation for a better medical treatment, minimizing the harmful effects. Many techniques and methods have been proposed to detect and quantify the radioinduced lesions in genetic material, among them, the micronucleus (MN) assay. In the present study, we proposed an improved in vitro micronucleus technique that is rapid, sensitive and with minor cell manipulations. Assays were carried out with human tumor cells (MCF-7) seeded (3x10 4 cells) in slides placed into Petri dishes. Adherent cells were maintained with RPMI medium, supplemented with fetal calf serum, 1 % antibiotics, cytochalasin B (2 μg/mL), and incubated at 37 deg C in the presence of 5% CO2 for 72h. Cells were pre-treated for 24h with aminoguanidine, a nitric oxide synthase inhibitor. Nitric oxide is an intracellular free-radical, involved in DNA double-strand break repair mechanisms. After incubation, adherent cells on slides were briefly fixed with paraformaldehyde and stained with acridine orange (100 μg/mL) for analysis through fluorescence microscopy. Dye fluorescence permitted accurate discrimination between nuclei and micronuclei (bright green) and cytoplasm (red), and made possible a faster counting of binucleated cells. Aminoguanidine (2 mM) induced significant increase (p< 0.05) in frequencies of binucleated cells with micronuclei and in the number of micronuclei per binucleated cell. Data showed that proposed modifications permit to understand an early aspect of NO inhibition and suggested an improved protocol to MN assays. (author)

  11. The cytokinesis-block micronucleus assay: a sensitive technique for measuring radiation-induced chromosome damage

    International Nuclear Information System (INIS)

    Fenech, M.; Morley, A.A.

    1987-01-01

    The sensitivity of the cytokinesis-block micronucleus assay was demonstrated by the detection in human lymphocytes of in vitro exposures of as low as 0.02 Gy of X-rays. To determine the suitability of this new method for measuring in vivo exposure to radiation the authors have performed initial longitudinal studies on (a) cancer patients undergoing partial body fractionated radiotherapy and (b) BALB-C mice following in vivo whole body irradiation with acute single doses of X-rays. The results for radiotherapy patients indicate that the dose fractions have an additive effect on the observed micronucleus frequency which appeared to decline following three months after completion of therapy. Results with irradiated mice showed a sharp increase in micronucleus frequency for splenocytes sampled immediately after treatment and the rate of decline in micronucleus frequency during the first week after treatment was dose-dependent. (author)

  12. Uncovering Aberrant Mutant PKA Function with Flow Cytometric FRET

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    Shin-Rong Lee

    2016-03-01

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

  13. 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

  14. Evaluation of river water genotoxicity using the piscine micronucleus test.

    Science.gov (United States)

    Ergene, Serap; Cavaş, Tolga; Celik, Ayla; Köleli, Nurcan; Aymak, Cemil

    2007-07-01

    The Berdan River, which empties into the Mediterranean Sea on the east coast of Turkey, receives discharges of industrial and municipal waste. In the present study, the in vivo piscine micronucleus (MN) test was used to evaluate the genotoxicity of water samples collected from different locations along the Berdan River. Nile tilapia (Oreochromis niloticus) were exposed in the laboratory for 2, 4, and 6 days, and micronuclei were evaluated in peripheral blood erythrocytes, gill cells, and caudal fin epithelial cells. A single dose of 5 mg/L cyclophosphamide was used as a positive control. In addition to micronuclei, nuclear abnormalities (NAs), such as binucleated cells and blebbed, notched, and lobed nuclei, were assessed in the erythrocytes, and chemical analyses were carried out to determine the amount of heavy metals in the water samples. MN and NA frequencies were significantly elevated (up to 2- to 3-fold) in fish exposed to river water samples taken downstream of potential discharges, and the elevated responses in gill and fin cells were related to the concentration of heavy metals in the water. MN frequencies (expressed as micronucleated cells/1,000 cells), in both treated and untreated fish, were greatest in gill cells (range: 0.80-3.70), and generally lower in erythrocytes (range: 0.50-2.80), and fin cells (range: 0.45-1.70). The results of this study indicate that the Berdan River is contaminated with genotoxic pollutants and that the genotoxicity is related to the discharge of wastes into the river water.

  15. Effects of electromagnetic fields induced from the visual display terminal on the micronucleus frequencies in tradescantia

    International Nuclear Information System (INIS)

    Shin, Hae Shik; Kim, Jin Kyu; Lee, Jin Hong

    2002-01-01

    Humans are exposed daily to electromagnetic fields (EMFs) originating from a variety of devices and VDT (Visual Display Terminal) workstations. This research was designed to examine the biological effects of electromagnetic fields from VDT using the Tradescantia- micronuclesus (Trad-MCN) bioassay. Tradescantia BNL 4430 clone was used to evaluate the influence of EMFs radiated from the VDT workstation. Trad-MCN assay is a cytogenetic test based on the formation of micronuclei that result from chromosome breakage in the meiotic pollen mother cells. No study has established unequivocally a causal relationship between EMFs and animals or plants. Fresh cuttings bearing young flower buds were exposed for 24hours in front of the VDT workstation. The cuttings were placed at 30, 50, 70, and 90cm distances from the workstations. The micronucleus were scored under a light microscope (400 x magnification). Three hundreds of tetrads were scored from each of the slides in every the experimental group. The frequencies expressed in terms of MCN / 100 tetrads. Trad-MCN frequencies were 8.93 ± 0.32, 11.2 ± 0.50, 7.67 ± 0.61, and 6.22 ± 1.78/ 100 tetrads at 30, 50, 70, and 90cm, respectively. In conclusion, EMFs from VDT give rise to damage the chromosome this plant. In addition, The results of the study indicate that Trad-MCN assay can detect chromosome damage due to EMFs from the electrical device workstation. In conclusion, the Trad-MCN assay is sensitive, reproducible, easy to perform, well standardized, inexpensive and undemanding in equipment

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

    Science.gov (United States)

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

    2012-01-01

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

  17. The buccal cytome and micronucleus frequency is substantially altered in Down's syndrome and normal ageing compared to young healthy controls

    International Nuclear Information System (INIS)

    Thomas, Philip; Harvey, Sarah; Gruner, Tini; Fenech, Michael

    2008-01-01

    The buccal micronucleus cytome assay was used to investigate biomarkers for DNA damage, cell death and basal cell frequency in buccal cells of healthy young, healthy old and young Down's syndrome cohorts. With normal ageing a significant increase in cells with micronuclei (P < 0.05, average increase +366%), karyorrhectic cells (P < 0.001, average increase +439%), condensed chromatin cells (P < 0.01, average increase +45.8%) and basal cells (P < 0.001, average increase +233%) is reported relative to young controls. In Down's syndrome we report a significant increase in cells with micronuclei (P < 0.001, average increase +733%) and binucleated cells (P < 0.001, average increase +84.5%) and a significant decrease in condensed chromatin cells (P < 0.01, average decrease -52%), karyolytic cells (P < 0.001, average decrease -51.8%) and pyknotic cells (P < 0.001, average decrease -75.0%) relative to young controls. These changes show distinct differences between the cytome profile of normal ageing relative to that for a premature ageing syndrome, and highlight the diagnostic value of the cytome approach for measuring the profile of cells with DNA damage, cell death and proportion of cells with proliferative potential (i.e., basal cells). Significant correlations amongst cell death biomarkers observed in this study were used to propose a new model of the inter-relationship of cell types scored within the buccal micronucleus cytome assay. This study validates the use of a cytome approach to investigate DNA damage, cell death and cell proliferation in buccal cells with ageing

  18. Screening potential genotoxic effect of aquatic plant extracts using the mussel micronucleus test

    Directory of Open Access Journals (Sweden)

    Bettina Eck-Varanka

    2016-01-01

    Full Text Available Objective: To assess the genotoxic potential of selected aquatic macrophytes: Ceratophyllum demersum L. (hornwort, family Ceratophyllaceae, Typha angustifolia L. (narrowleaf cattail, family Typhaceae, Stratiotes aloides L. (water soldier, family Butomaceae, and Oenanthe aquatica (L. Poir. (water dropwort, family Umbelliferae. Methods: For genotoxicity assessment, the mussel micronucleus test was applied. Micronucleus frequency was determined from the haemolymph of Unio pictorum L. (painter’s mussel. In parallel, total and hydrolisable tannin contents were determined. Results: All plant extracts elucidated significant mutagenic effect. Significant correlation was determined between tannin content and mutagenic capacity. Conclusions: The significant correlation between genotoxicity as expressed by micronucleus frequency and tannin content (both total and hydrolisable tannins indicate that tannin is amongst the main compounds being responsible for the genotoxic potential. It might be suggested that genotoxic capacity of these plants elucidate a real ecological effect in the ecosystem.

  19. Radiation dose-response relationship of micronucleus occurrence in pollen mother cells of tradescantia

    International Nuclear Information System (INIS)

    Kim, Jin Kyu; Kim, Yeon Ku; Song, Hi Sup

    1999-01-01

    This study was carried out to investigate the radiation dose-response of micronucleus frequencies in Tradescantia pollen mother cells. The number of micronuclei increased in the tetrads as a result of chromosome deletion after irradiation. The maximal frequency of micronucleus showed a good dose-response relationship in the range of dose 0∼50 cGy. On the basis of the relationship, a dose of 1 cGy resulted in two additional micronuclei in 100 tetrads. The radiation dose-response relationship of micronucleus occurrence is prerequisite to biological monitoring of radiation and can be modified for biological risk assessment of toxicants, and to safety test of water or soil integrity

  20. Leukocytospermia and sperm preparation - a flow cytometric study

    Directory of Open Access Journals (Sweden)

    Perticarari Sandra

    2009-01-01

    Full Text Available Abstract Background Leukocytes represent the predominant source of reactive oxygen species both in seminal plasma and in sperm suspensions and have been demonstrated to negatively influence sperm function and fertilization rate in assisted reproduction procedures. Peroxidase test is the standard method recommended by WHO to detect semen leukocytes but it may be inaccurate. The aims of this study were (i to compare the efficiency of swim-up and density-gradient centrifugation techniques in removing seminal leukocytes, (ii to examine the effect of leukocytes on sperm preparation, and (iii to compare flow cytometry and peroxidase test in determining leukocyte concentration in semen using a multiparameter flow cytometric method. Methods Semen samples from 126 male partners of couples undergoing infertility investigations were analyzed for leukocytospermia using standard optical microscopy and flow cytometry. Sixty-nine out of 126 samples were also processed using simultaneously the swim-up and density-gradient centrifugation techniques. A multiparameter flow cytometric analysis to assess simultaneously sperm concentration, sperm viability, sperm apoptosis, and leukocyte concentration was carried out on neat and prepared sperm. Results Both sperm preparation methods removed most seminal leukocytes. However, the concentration of leukocytes was significantly lower after swim-up compared to that after density-gradient centrifugation preparation. Leukocytes concentration, either initial or in prepared fractions, was not correlated with sperm parameters (optical microscopy and flow cytometry parameters after semen processing. There was no correlation between leukocyte concentration in the ejaculate and sperm recovery rate, whereas a significant correlation was found between the concentration of the residual leukocytes in prepared fractions and viable sperm recovery rate. Although the overall concordance between the flow cytometry and the optical

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

    International Nuclear Information System (INIS)

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

    1988-01-01

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

  2. 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

  3. 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.

  4. 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.

  5. Absence of micronucleus formation in CHO-K1 cells cultivated in platelet lysate enriched medium.

    Science.gov (United States)

    Bernardi, Martina; Adami, Valentina; Albiero, Elena; Madeo, Domenico; Rodeghiero, Francesco; Astori, Giuseppe

    2014-03-01

    Human platelet lysate (PL) represents an effective substitute of fetal bovine serum (FBS) for mesenchymal stromal cell (MSC) cultivation. Compared to FBS, PL favors MSC proliferation significantly shortening the population doubling time and avoiding the risks related to the use of animal derivatives. Growth factors contained in the platelets are released upon platelet disruption following freezing/thawing cycles or as we have recently described by using ultrasound. We have investigated whether the increased cell proliferation achieved by using PL could induce mitotic stress and whether the potential formation of free radicals during PL production by ultrasound could cause chromosomal instability in mammalian cells. We have applied an image analysis assisted high content screening (HCS) in vitro micronucleus assay in the Chinese Hamster Ovarian K1 (CHO-K1) rodent mammalian cell line. PL was produced by sonication; for the micronucleus assay, CHO-K1 cells were exposed to increasing concentrations of PL. Cytokinesis was blocked by cytochalasin B, nuclei were stained with bisbenzimide and images were acquired and analyzed automatically using an HCS system, both with a 20× and a 10× objective. Our results suggest that growth stimulus induced by the use of PL did not significantly increase micronucleus formation in CHO-K1 cells compared to negative control. Micronucleus testing in conjunction with HCS could represent a valid tool to evaluate the safety of ancillary materials used in the production of cell-based medicinal products. Copyright © 2013 Elsevier GmbH. All rights reserved.

  6. 40 CFR 798.5395 - In vivo mammalian bone marrow cytogenetics tests: Micronucleus assay.

    Science.gov (United States)

    2010-07-01

    ... Genetic Toxicity § 798.5395 In vivo mammalian bone marrow cytogenetics tests: Micronucleus assay. (a... and documented with data, only this one time point need be sampled. (ii) If a repeated treatment... slides, spread as a smear and stained. (2) Analysis. Slides shall be coded before microscopic analysis...

  7. Association between micronucleus frequency and cervical intraepithelial neoplasia grade in Thinprep cytological test and its significance.

    Science.gov (United States)

    Shi, Yong-Hua; Wang, Bo-Wei; Tuokan, Talaf; Li, Qiao-Zhi; Zhang, Ya-Jing

    2015-01-01

    A micronucleus is an additional small nucleus formed due to chromosomes or chromosomal fragments fail to be incorporated into the nucleus during cell division. In this study, we assessed the utility of micronucleus counting as a screening tool in cervical precancerous lesions in Thinprep cytological test smears under oil immersion. High risk HPV was also detected by hybrid capture-2 in Thinprep cytological test smears. Our results showed that micronucleus counting was significantly higher in high-grade squamous intraepithelial lesion (HSIL) and invasive carcinoma cases compared to low-grade squamous intraepithelial lesion (LSIL) and non-neoplastic cases. Receiver operating characteristic (ROC) curve analysis revealed that micronucleus counting possessed a high degree of sensitivity and specificity for identifying HSIL and invasive carcinoma. Cut-off of 7.5 for MN counting gave a sensitivity of 89.6% and a specificity of 66.7% (P = 0.024 and AUC = 0.892) for detecting HSIL and invasive carcinoma lesions. Multiple linear regression analysis showed that only HSIL and invasive cancer lesions not age, duration of marital life and number of pregnancy are significantly associated with MN counting. The positive rate of high risk HPV was distinctly higher in LSIL, HSIL and invasive cancer than that in non-neoplstic categories. In conclusions, MN evaluation may be viewed as an effective biomarker for cervical cancer screening. The combination of MN count with HPV DNA detection and TCT may serve as an effective means to screen precancerous cervical lesions in most developing nations.

  8. Significance of micronucleus in cervical intraepithelial lesions and carcinoma

    Directory of Open Access Journals (Sweden)

    B N Gayathri

    2012-01-01

    Conclusions: MN scoring on the epithelial cells of cervix could be used as a biomarker in cancer screening. This is an easy, simple, reliable, reproducible and objective test which can be performed on routinely stained pap smears.

  9. 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

  10. Radiosensitiviness of blood lymphocytes from skin cancer patients and healthy volunteers as determined by micronucleus assay

    International Nuclear Information System (INIS)

    Lohmann, Tania Helena Ochi.

    1995-01-01

    Cancer, a major death cause in developed countries, has been related to somatic mutations that could be detected by cytogenetic analysis. Among the tools used in these tests, the micronucleus assay has been largely applied at population surveillance, biological dosimetry and early detection of groups with higher risks to developing cancers. In this study, we analysed the chromosome susceptibility of blood lymphocytes from basocellular skin cancer patients and healthy volunteers. The cytogenetic analysis was performed by a micronucleus assay, using progressive doses of ionizing radiation from a 60 Co source as mutagen. Briefly, the blood lymphocytes were irradiated in vitro, as processed by the cytokinesis-blocked method. The micronucleus frequency and distribution, cell cycle kinetics, nucleation index and dose-response relationship were determined in each patient. The results showed that the basocellular skin cancer patients lymphocytes presented higher spontaneous micronucleus frequency as compared with those from healthy young volunteers but lower than healthy now young volunteers . The radiation-induced micronucleus analysis showed that the basocellular skin cancer patients' lymphocytes presented similar proportion of damage lymphocytes as compared with those from healthy volunteers. Nevertheless, the magnitude of this damage was higher in this group with doses. Higher than 400 c Gy, which was not occurred in healthy volunteers. Cell cycle kinetics, as determined by the nucleation index, was lower in basocellular skin cancer patients as compared with healthy volunteers, indicating a more slow cell cycle. Our data showed that the lymphocytes from carcinoma basocellular patients were more radiosensitive as compared with those form healthy volunteers. (author). 159 refs., 21 figs., 16 tabs

  11. Dose-rate effects for apoptosis and micronucleus formation in gamma-irradiated human lymphocytes

    International Nuclear Information System (INIS)

    Boreham, D.R.; Dolling, J.-A.; Maves, S.R.; Siwarungsun, N.; Mitchel, R.E.J.

    2000-01-01

    We have compared dose-rate effects for γ-radiation-induced apoptosis and micronucleus formation in human lymphocytes. Long-term assessment of individual radiation-induced apoptosis showed little intraindividual variation but significant interindividual variation. The effectiveness of radiation exposure to cause apoptosis or micronucleus formation was reduced by low-dose-rate exposures, but the reduction was apparent at different dose rates for these two end points. Micronucleus formation showed a dose-rate effect when the dose rate was lowered to 0.29 cGy/min, but there was no accompanying cell cycle delay. A further increase in the dose-rate effect was seen at 0.15 cGy/min, but was now accompanied by cell cycle delay. There was no dose-rate effect for the induction of apoptosis until the dose rate was reduced to 0.15 cGy/min, indicating that the mechanisms or signals for processing radiation-induced lesions for these two end points must be different at least in part. There appear to be two mechanisms that contribute to the dose-rate effect for micronucleus formation. One of these does not affect binucleate cell frequency and occurs at dose rates higher than that required to produce a dose-rate effect for apoptosis, and one affects binucleate cell frequency, induced only at the very low dose rate which coincidentally produces a dose-rate effect for apoptosis. Since the dose rate at which cells showed reduced apoptosis as well as a further reduction in micronucleus formation was very low, we conclude that the processing of the radiation-induced lesions that induce apoptosis, and some micronuclei, is very slow in quiescent and PHA-stimulated lymphocytes, respectively. (author)

  12. Dose-rate effects for apoptosis and micronucleus formation in gamma-irradiated human lymphocytes

    Energy Technology Data Exchange (ETDEWEB)

    Boreham, D.R.; Dolling, J.-A.; Maves, S.R. [Atomic Energy of Canada Limited, Chalk River, Ontario (Canada); Siwarungsun, N. [Chulalongkorn Univ., Bangkok (Thailand); Mitchel, R.E.J. [Atomic Energy of Canada Limited, Chalk River, Ontario (Canada)

    2000-07-01

    We have compared dose-rate effects for {gamma}-radiation-induced apoptosis and micronucleus formation in human lymphocytes. Long-term assessment of individual radiation-induced apoptosis showed little intraindividual variation but significant interindividual variation. The effectiveness of radiation exposure to cause apoptosis or micronucleus formation was reduced by low-dose-rate exposures, but the reduction was apparent at different dose rates for these two end points. Micronucleus formation showed a dose-rate effect when the dose rate was lowered to 0.29 cGy/min, but there was no accompanying cell cycle delay. A further increase in the dose-rate effect was seen at 0.15 cGy/min, but was now accompanied by cell cycle delay. There was no dose-rate effect for the induction of apoptosis until the dose rate was reduced to 0.15 cGy/min, indicating that the mechanisms or signals for processing radiation-induced lesions for these two end points must be different at least in part. There appear to be two mechanisms that contribute to the dose-rate effect for micronucleus formation. One of these does not affect binucleate cell frequency and occurs at dose rates higher than that required to produce a dose-rate effect for apoptosis, and one affects binucleate cell frequency, induced only at the very low dose rate which coincidentally produces a dose-rate effect for apoptosis. Since the dose rate at which cells showed reduced apoptosis as well as a further reduction in micronucleus formation was very low, we conclude that the processing of the radiation-induced lesions that induce apoptosis, and some micronuclei, is very slow in quiescent and PHA-stimulated lymphocytes, respectively. (author)

  13. Biological Dosimetry Using Micronucleus Assay in Simulated Partial-Body Exposure to Ionizing Radiation

    Directory of Open Access Journals (Sweden)

    S. Purnami

    2017-08-01

    Full Text Available In radiation accidents, it is common that only several parts of the body are exposed to radiation. As a consequence there is a mixture of exposed and unexposed lymphocytes in peripheral blood cells of the samples. This phenomenon will cause the dose value estimated using the exposed lymphocytes to be lower than the actual dose. In this study, an assessment of partial body exposures using micronucleus assay by estimating the partial body dose and fraction of irradiated blood was conducted. An optimal D0 value also has been determined in this study to estimate the fraction of irradiated cells. Peripheral blood lymphocytes (PBLs from three healthy donors were irradiated in vitro with 2 Gy of X-rays. Partial radiation exposure was simulated by mixing the irradiated and non-irradiated blood in different proportions. The proportions of mixtures of blood samples irradiated in vitro were 5, 10, 15, 20, and 30 %. Blood samples were then cultured and harvested based on micronuclei assay protocol. At least 2000 binucleated cells with well-preserved cytoplasm were scored for the MN frequency. Dose Estimate 5.1 software was used to calculate the dispersion index (σ2/y and normalized unit of this index (U in each proportion of bloods. The fractions of irradiated cells were calculated with CABAS (Chromosomal Aberration Calculation Software for several different D0 values (2.7; 3.8; 5.4. The results showed that D0 value at 5.4 gave the closest results to the actual proportion of irradiated bloods, while for the dose estimation the estimated doses value from all proportions in all donors were higher than the actual dose. The factor that may cause this phenomenon was that the dose response calibration curve used to predict the radiation dose was not constructed in the laboratory used. Overall it can be concluded that a biodosimetry using MN assay can be used to estimate the radiation dose in partial body exposure. In order to establish a biodosimetry using MN

  14. State of the Art High-Throughput Approaches to Genotoxicity: Flow Micronucleus, Ames II, GreenScreen and Comet

    Science.gov (United States)

    State of the Art High-Throughput Approaches to Genotoxicity: Flow Micronucleus, Ames II, GreenScreen and Comet (Presented by Dr. Marilyn J. Aardema, Chief Scientific Advisor, Toxicology, Dr. Leon Stankowski, et. al. (6/28/2012)

  15. Analysis of resveratrol and radiation effects in lung cancer cells by micronucleus assay

    Energy Technology Data Exchange (ETDEWEB)

    Moreno, Carolina S.; Santos, Dymes R.A.; Vieira, Daniel P.; Rogero, Sizue O.; Rogero, Jose R., E-mail: carolina_sm@hotmail.com [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil); Sakuraba, Roberto K.; Weltman, Eduardo [Hospital Israelita Albert Einstein, Sao Paulo, SP (Brazil); Cruz, Aurea S.; Santos, Rezolina P. [Instituto Adolfo Lutz, Sao Paulo, SP (Brazil)

    2015-07-01

    Mucoepidermoid lung carcinoma is frequently manifested by obstructive trachea symptoms. Radiation and drugs combinations are commonly used in the lung cancer treatment. Currently there is a strong tendency to develop therapeutic strategies focused at the administration of high potential compounds to improve the ionizing radiation treatments, so as to increase the radiation effects on tumor cell while minimizing these effects to surrounding normal tissues. Resveratrol is a polyphenolic phytoalexin compound present in wines and several plants. This compound has a broad spectrum of biological activities such as antioxidant, anticarcinogenic, and induction of cell cycle arrest effects. Analysis of biological effects of ionizing radiation in the presence of resveratrol in different cell cultures has been the subject of many studies. To verify the genotoxic effects in cells exposed to ionizing radiation many methods have been proposed. The cytokinesis-block micronucleus technique is one of the preferred methods. The main of this study was to detect and quantify radioinduced DNA damage in mucoepidermoid lung carcinoma cells (NCI-H292) by cytokinesis-block micronucleus technique using cytocalasin-B. The cell culture was irradiated at a single fraction from a TrueBeam® linear accelerator (0, 0.8, 5, and 10 Gy), in the absence or presence of different resveratrol concentrations (0, 15, 30, and 60 μM). The results showed that resveratrol (15 and μM) induced significant increase frequency (p<0.05) of micronucleus formation in NCI-H292 cell culture non-irradiated and exposed at 5 Gy dose. Moreover, resveratrol (30 μM) induced micronucleus formation at 0.8 Gy dose. (author)

  16. Analysis of resveratrol and radiation effects in lung cancer cells by micronucleus assay

    International Nuclear Information System (INIS)

    Moreno, Carolina S.; Santos, Dymes R.A.; Vieira, Daniel P.; Rogero, Sizue O.; Rogero, Jose R.; Sakuraba, Roberto K.; Weltman, Eduardo; Cruz, Aurea S.; Santos, Rezolina P.

    2015-01-01

    Mucoepidermoid lung carcinoma is frequently manifested by obstructive trachea symptoms. Radiation and drugs combinations are commonly used in the lung cancer treatment. Currently there is a strong tendency to develop therapeutic strategies focused at the administration of high potential compounds to improve the ionizing radiation treatments, so as to increase the radiation effects on tumor cell while minimizing these effects to surrounding normal tissues. Resveratrol is a polyphenolic phytoalexin compound present in wines and several plants. This compound has a broad spectrum of biological activities such as antioxidant, anticarcinogenic, and induction of cell cycle arrest effects. Analysis of biological effects of ionizing radiation in the presence of resveratrol in different cell cultures has been the subject of many studies. To verify the genotoxic effects in cells exposed to ionizing radiation many methods have been proposed. The cytokinesis-block micronucleus technique is one of the preferred methods. The main of this study was to detect and quantify radioinduced DNA damage in mucoepidermoid lung carcinoma cells (NCI-H292) by cytokinesis-block micronucleus technique using cytocalasin-B. The cell culture was irradiated at a single fraction from a TrueBeam® linear accelerator (0, 0.8, 5, and 10 Gy), in the absence or presence of different resveratrol concentrations (0, 15, 30, and 60 μM). The results showed that resveratrol (15 and μM) induced significant increase frequency (p<0.05) of micronucleus formation in NCI-H292 cell culture non-irradiated and exposed at 5 Gy dose. Moreover, resveratrol (30 μM) induced micronucleus formation at 0.8 Gy dose. (author)

  17. Assessment of the genotoxicity of 137Cs radiation using Vicia-micronucleus, Tradescantia-micronucleus and Tradescantia-stamen-hair mutation bioassays.

    Science.gov (United States)

    Minouflet, Marion; Ayrault, Sophie; Badot, Pierre-Marie; Cotelle, Sylvie; Ferard, Jean-François

    2005-01-01

    Since the middle of the 20th century, ionizing radiations from radioactive isotopes including 137Cs have been investigated to determine their genotoxic impact on living organisms. The present study was designed to compare the effectiveness of three plant bioassays to assess DNA damage induced by low doses of 137Cs: Vicia-micronucleus test (Vicia-MCN), Tradescantia-micronucleus test (Trad-MCN) and Tradescantia-stamen-hair mutation test (Trad-SH) were used. Vicia faba (broad bean) and Tradescantia clone 4430 (spiderwort) were exposed to 137Cs according to different scenarios: external and internal (contamination) irradiations. Experiments were conducted with various levels of radioactivity in solution or in soil, using solid or liquid 137Cs sources. The three bioassays showed different sensitivities to the treatments. Trad-MCN appeared to be the most sensitive test (significative response from 1.5 kBq/200 ml after 30 h of contamination). Moreover, at comparable doses, internal irradiations led to larger effects for the three bioassays. These bioassays are effective tests for assessing the genotoxic effects of radioactive 137Cs pollution.

  18. Assessment of the genotoxicity of 137Cs radiation using Vicia-micronucleus, Tradescantia-micronucleus and Tradescantia-stamen-hair mutation bioassays

    International Nuclear Information System (INIS)

    Minouflet, Marion; Ayrault, Sophie; Badot, Pierre-Marie; Cotelle, Sylvie; Ferard, Jean-Francois

    2005-01-01

    Since the middle of the 20th century, ionizing radiations from radioactive isotopes including 137 Cs have been investigated to determine their genotoxic impact on living organisms. The present study was designed to compare the effectiveness of three plant bioassays to assess DNA damage induced by low doses of 137 Cs: Vicia-micronucleus test (Vicia-MCN), Tradescantia-micronucleus test (Trad-MCN) and Tradescantia-stamen-hair mutation test (Trad-SH) were used. Vicia faba (broad bean) and Tradescantia clone 4430 (spiderwort) were exposed to 137 Cs according to different scenarios: external and internal (contamination) irradiations. Experiments were conducted with various levels of radioactivity in solution or in soil, using solid or liquid 137 Cs sources. The three bioassays showed different sensitivities to the treatments. Trad-MCN appeared to be the most sensitive test (significative response from 1.5 kBq/200 ml after 30 h of contamination). Moreover, at comparable doses, internal irradiations led to larger effects for the three bioassays. These bioassays are effective tests for assessing the genotoxic effects of radioactive 137 Cs pollution

  19. Evaluation of 10 aliphatic halogenated hydrocarbons in the mouse bone marrow micronucleus test.

    Science.gov (United States)

    Crebelli, R; Carere, A; Leopardi, P; Conti, L; Fassio, F; Raiteri, F; Barone, D; Ciliutti, P; Cinelli, S; Vericat, J A

    1999-03-01

    Ten halogenated aliphatic hydrocarbons (carbon tetrachloride, 1-chlorohexane, 2,3-dichlorobutane, 1,2-dichloroethane, 1,2-dichloroethylene, 1,3-dichloropropane, hexachloroethane, 1,1,2-trichloroethane, 1,2,3-trichloropropane and 1,1,3-trichloropropene), previously assayed in genetic assays in fungi, were evaluated in the mouse bone marrow micronucleus test in order to assess their genotoxicity in vivo. All chemicals were administered once i.p. at 40 and 70-80% of their respective LD50 to male and female CD-1 mice, 24 and 48 h before killing. All treatments produced evident clinical symptoms, but no marked depression of bone marrow proliferation. No statistically significant increases in the incidence of micronucleated polychromatic erythrocytes over the control values were observed at any sampling time with any of the 10 halogenated hydrocarbons assayed. The comparison of the results obtained in this study with the findings provided by in vitro micronucleus assays on the same chemicals, reported by other authors, indicate that mouse bone marrow is weakly sensitive to the genotoxic effects induced by halogenated hydrocarbons in other test systems. This suggests that the role of such an assay in carcinogen screening may be questionable for this chemical class. An examination of mouse bone marrow micronucleus test results with the halogenated aliphatic hydrocarbons classified as carcinogens by IARC supports this conclusion.

  20. Quantifying Distribution of Flow Cytometric TCR-Vβ Usage with Economic Statistics.

    Directory of Open Access Journals (Sweden)

    Kornelis S M van der Geest

    Full Text Available Measuring changes of the T cell receptor (TCR repertoire is important to many fields of medicine. Flow cytometry is a popular technique to study the TCR repertoire, as it quickly provides insight into the TCR-Vβ usage among well-defined populations of T cells. However, the interpretation of the flow cytometric data remains difficult, and subtle TCR repertoire changes may go undetected. Here, we introduce a novel means for analyzing the flow cytometric data on TCR-Vβ usage. By applying economic statistics, we calculated the Gini-TCR skewing index from the flow cytometric TCR-Vβ analysis. The Gini-TCR skewing index, which is a direct measure of TCR-Vβ distribution among T cells, allowed us to track subtle changes of the TCR repertoire among distinct populations of T cells. Application of the Gini-TCR skewing index to the flow cytometric TCR-Vβ analysis will greatly help to gain better understanding of the TCR repertoire in health and disease.

  1. 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 orthogona...

  2. 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

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

    NARCIS (Netherlands)

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

    1993-01-01

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

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

    DEFF Research Database (Denmark)

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

    2004-01-01

    were defined: region 1 includes bacteria mainly associated with poor hygiene, region 2 includes psychrotrophic hygiene bacteria and region 3 includes bacteria mainly related to mastitis. The ability of the flow cytometric technique to predict the main cause of elevated bacterial counts on routine...

  5. Variability in micronucleus induction with different mutagens applied to several species of fish

    Directory of Open Access Journals (Sweden)

    Cesar Koppe Grisolia

    2000-03-01

    Full Text Available Fish are often used for screening genotoxicity of water. For such programs, a knowledge of the sensitivity to clastogens, spontaneous micronucleus frequency and cell cycle kinetics of the target tissue is necessary. To investigate the pattern of inter-specific sensitivity to micronucleus induction three species of fish, Tilapia rendalli, Oreochromis niloticus and Cyprinus carpio, were exposed to the clastogens bleomycin (BLM, cyclophosphamide (CP, 5-fluorouracil (5-FU, and mitomycin C (MMC. The binucleate/mononucleate ratio in peripheral erythrocytes exposed to cytochalasin B was also used to evaluate the time-dependent response of micronucleus formation during hematopoesis in the kidney and the micronucleus peak in peripheral erythrocytes. Micronucleus frequencies induced by CP were significantly greater than their respective controls for the three fish species throughout all treatment periods. During the whole evaluation period (30 days CP was also the most effective clastogen. In general, until the 14th day of evaluation period T. rendalii was the most sensitive species to clastogens. No difference in micronucleus frequencies among species was observed in the 4th evaluation (at the 30th day. A micronucleus peak was observed at the 7th day after treatment. After the 14th day the frequencies were stabilized. The cytochalasin B experiment was carried out to demonstrate that micronuclei induced in the young kidney erythrocyte cells were detected in the circulating blood 2-4 days later.Este estudo fez uma avaliação da indução de micronúcleos em eritrócitos de sangue periférico de peixes Tilapia rendalli, Oreochromis niloticus e Cyprinus carpio após o tratamento com mitomicina C, ciclofosfamida, 5-fluorouracil e bleomicina. Foram colhidas amostras periódicas de sangue com 2, 7, 14 e 30 dias após o tratamento único. Os tratamentos com citocalasina B tiveram como objetivo analisar as proporções entre células binucleadas

  6. Micronucleus test in mice fed on irradiated whole diet

    International Nuclear Information System (INIS)

    Reddy, P.P.; Reddi, O.S.; Pentiah, P.R.; Rani, M.V.U.; Devi, K.R.; Goud, S.N.

    1981-01-01

    Eight week old Swiss albino male mice were fed on freshly irradiated or unirradiated whole diet for one week. (Exposure was to 75 or 200 kR γ rays from a 1000 Ci 60 Co γ source at a dose rate of 584 R/min.) On the seventh day, six hours after feeding, the mice were killed and bone marrow preparations were made by the Schmid technique. From each group three animals were taken and from each animal 2000 polychromatic and normochromatic erythrocytes were scored. It was evident from the data obtained that the irradiated whole diet failed to induce any significant increase in the incidence of micronuclei in polychromatic erythrocytes. Similarly, there was no significant increase in the frequency of micronuclei in normochromatic erythrocytes when compared with control data. The polychromatic to normochromatic ratio was also unaffected. The diet consisted of wheat flour (60%). groundnut cake (20%), fish meal (8%), Bengal gram flour (8%), dried yeast (3%), salt/mineral mixture (1%) and traces of vitamins. (U.K.)

  7. The buccal cytome and micronucleus frequency is substantially altered in Down's syndrome and normal ageing compared to young healthy controls

    Energy Technology Data Exchange (ETDEWEB)

    Thomas, Philip [CSIRO Human Nutrition, PO Box 10041, Adelaide BC, Adelaide, SA 5000 (Australia); Discipline of Physiology, School of Molecular and Biomedical Sciences, The University of Adelaide, Adelaide, SA 5005 (Australia)], E-mail: philip.thomas@csiro.au; Harvey, Sarah; Gruner, Tini [Southern Cross University, PO Box 157, Lismore, NSW 2480 (Australia); Fenech, Michael [CSIRO Human Nutrition, PO Box 10041, Adelaide BC, Adelaide, SA 5000 (Australia)], E-mail: michael.fenech@csiro.au

    2008-02-01

    The buccal micronucleus cytome assay was used to investigate biomarkers for DNA damage, cell death and basal cell frequency in buccal cells of healthy young, healthy old and young Down's syndrome cohorts. With normal ageing a significant increase in cells with micronuclei (P < 0.05, average increase +366%), karyorrhectic cells (P < 0.001, average increase +439%), condensed chromatin cells (P < 0.01, average increase +45.8%) and basal cells (P < 0.001, average increase +233%) is reported relative to young controls. In Down's syndrome we report a significant increase in cells with micronuclei (P < 0.001, average increase +733%) and binucleated cells (P < 0.001, average increase +84.5%) and a significant decrease in condensed chromatin cells (P < 0.01, average decrease -52%), karyolytic cells (P < 0.001, average decrease -51.8%) and pyknotic cells (P < 0.001, average decrease -75.0%) relative to young controls. These changes show distinct differences between the cytome profile of normal ageing relative to that for a premature ageing syndrome, and highlight the diagnostic value of the cytome approach for measuring the profile of cells with DNA damage, cell death and proportion of cells with proliferative potential (i.e., basal cells). Significant correlations amongst cell death biomarkers observed in this study were used to propose a new model of the inter-relationship of cell types scored within the buccal micronucleus cytome assay. This study validates the use of a cytome approach to investigate DNA damage, cell death and cell proliferation in buccal cells with ageing.

  8. Does the recommended lymphocyte cytokinesis-block micronucleus assay for human biomonitoring actually detect DNA damage induced by occupational and environmental exposure to genotoxic chemicals?

    Science.gov (United States)

    Speit, Günter

    2013-07-01

    This commentary challenges the paradigm that the cytokinesis-block micronucleus assay (CBMN assay) with cultured human lymphocytes, as it is performed currently, is a sensitive and useful tool for detecting genotoxic effects in populations exposed occupationally or environmentally to genotoxic chemicals. Based on the principle of the assay and the available data, increased micronucleus (MN) frequencies in binucleated cells (BNC) are mainly due to MN produced in vitro during the cultivation period (i.e. MN produced in vivo do not substantially contribute to the MN frequency measured in BNC). The sensitivity of the assay for the detection of induced MN in BNC after an in vivo exposure to a genotoxic chemical is limited because cytochalasin B (Cyt-B) is added relatively late during the culture period and, therefore, the BNC that are scored do not always represent cells that have completed one cell cycle only. Furthermore, this delay means that damaged cells can be eliminated by apoptosis and/or that DNA damage induced in vivo can be repaired prior to the production of a MN in the presence of Cyt-B. A comparison with the in vitro CBMN assay used for genotoxicity testing leads to the conclusion that it is highly unlikely that DNA damage induced in vivo is the cause for increased MN frequencies in BNC after occupational or environmental exposure to genotoxic chemicals. This commentary casts doubt on the usefulness of the CBMN assay as an indicator of genotoxicity in human biomonitoring and questions the relevance of many published data for hazard identification and risk assessment. Thus, it seems worthwhile to reconsider the use of the CBMN assay as presently conducted for the detection of genotoxic exposure in human biomonitoring.

  9. Health assessment of gasoline and fuel oxygenate vapors: micronucleus and sister chromatid exchange evaluations.

    Science.gov (United States)

    Schreiner, Ceinwen A; Hoffman, Gary M; Gudi, Ramadevi; Clark, Charles R

    2014-11-01

    Micronucleus and sister chromatid exchange (SCE) tests were performed for vapor condensate of baseline gasoline (BGVC), or gasoline with oxygenates, methyl tert-butyl ether (G/MTBE), ethyl tert butyl ether (G/ETBE), t-amyl methyl ether (G/TAME), diisopropyl ether (G/DIPE), t-butyl alcohol (TBA), or ethanol (G/EtOH). Sprague Dawley rats (the same 5/sex/group for both endpoints) were exposed to 0, 2000, 10,000, or 20,000mg/m(3) of each condensate, 6h/day, 5days/week over 4weeks. Positive controls (5/sex/test) were given cyclophosphamide IP, 24h prior to sacrifice at 5mg/kg (SCE test) and 40mg/kg (micronucleus test). Blood was collected from the abdominal aorta for the SCE test and femurs removed for the micronucleus test. Blood cell cultures were treated with 5μg/ml bromodeoxyuridine (BrdU) for SCE evaluation. No significant increases in micronucleated immature erythrocytes were observed for any test material. Statistically significant increases in SCE were observed in rats given BGVC alone or in female rats given G/MTBE. G/TAME induced increased SCE in both sexes at the highest dose only. Although DNA perturbation was observed for several samples, DNA damage was not expressed as increased micronuclei in bone marrow cells. Inclusion of oxygenates in gasoline did not increase the effects of gasoline alone or produce a cytogenetic hazard. Copyright © 2014 Elsevier Inc. All rights reserved.

  10. Study on ionizing radiation to the workers' lymphocyte micronucleus rate and chromosome aberrations

    International Nuclear Information System (INIS)

    Li Jianhua; Wang Linchao; He Wei

    2007-01-01

    Objective: To study lymphocyte genetic material of an iron and steel enterprise workers exposed to the ionizing radiation, find out measures to protect their health and reduce ionizing radiation occupation harm. Methods: 342 workers were choseh as the exposed group who worked in an iron and steel enterprise in the beam installment operation, to examine their circumference blood lymphocyte micronucleus rate and the chromosome aberrations, simultaneously select 280 chefs as the control group, The irradiation dosage was determined and statistical analysis was carded out wich the consideration of their length of work and differences in work post. Results: Exposed group: the micronucleus rate masculine gender (MNR), 4 people, the masculine gender pick out rate is 12.87%. The chromosome aberration factor masculine gender (CAF), 12 people, the masculine rate is 3.51%. Control group: MNR 3 people, the asculine gender pick out rate is 1.07%; CAF 2 people, masculine gender rate is 0.72%. Comparing the two groups, every item has the significant difference. Workers in is the exposed group workers have the average exposure dose of 6.73mSv/a, MNR,CAF are illuminated to the dosage have a positive line correlation. They become increased as the job lenght prolongs. The nucleon name, the material calculation and the medical X-radial are responsible for the highest ratio. Conclusion: In iron and steel enterprises, long-time ionizing radiation can cause the workers' circumference blood lymphocyte micronucleus rate and the chromosome aberrations obvious to rise. The beam protection measures strengthened so as to reduce the harms to workers. (authors)

  11. Cytochalasin-b micronucleus test of peripheral blood lymphocytes of Kozloduy NPP workers

    International Nuclear Information System (INIS)

    Hadjidekova, V.; Hristova, R.; Atanassova, P.; Stainova, A.; Popova, L.

    2006-01-01

    Full text: The cytokinesis-block micronucleus assay in peripheral blood lymphocytes was applied to evaluate occupational radiation exposure of 65 nuclear power plant workers. Blood samples were collected from 43 workers aged between 32-54 years, mean age 41,7 years. The accumulated radiation doses for each subject varied between 7,9 - 766,4 mSv, mean level of the whole group is 237,78 mSv. Controls were 22 healthy individuals, (13 male and 9 female), aged between 27-52 years, mean age 38,8 years, selected from the administrative staff. All subjects participating in this study were interviewed concerning health status, professional history, smoking habit and other aspects relevant to the study. At least 1000 binucleated cells were analysed per person. The detected frequencies of micronuclei in the control group were ranged between 4.0 and 23.5 per 1000 binucleated cells, with the average incidence yield of 12.16 ±5.59 %. The mean group value of the frequency of micronuclei in peripheral lymphocytes of exposed workers was found to be 18.46±6.72 % in 1000 cells. The difference between the mean frequency of micronuclei in the group of exposed subjects and the control group was statistically significant (P<0,001). The correlation coefficient for duration of employment and accumulated doses is 0,30 (P<0,05). After 1,5 Gy in vitro irradiation of peripheral blood from investigated workers and controls a decreased radiosensitivity of NPP workers is detected using micronucleus assay. Decreased radiosensitivity of the professionally exposed workers could be due to the phenomenon of adaptive response. Micronucleus assay in peripheral blood lymphocytes is useful approach in cytogenetic monitoring of occupationally exposed nuclear industry workers

  12. Nucleoplasmic bridges are a sensitive measure of chromosome rearrangement in the cytokinesis-block micronucleus assay

    International Nuclear Information System (INIS)

    Fenech, M.; Umegaki, K.

    2003-01-01

    Full text: We have performed experiments using the WIL2-NS human B-lymphoblastoid cell line and primary human lymphocytes to (a) determine the importance of including measurements of nucleoplasmic bridges (NPB) in the cytokinesis-block micronucleus (CBMN) assay and (b) provide evidence that NPB originate from dicentric chromosomes and centric ring chromosomes. In addition we describe theoretical models that explain how dicentric chromosomes and centric ring chromosomes may result in the formation of NPB at anaphase. The results with WIL2-NS showed that it was possible to distinguish genotoxic effects induced by different oxidizing agents in terms of the NPB/micronucleus frequency ratio. The results with lymphocytes indicated a strong correlation (a) between NPB, centric ring chromosomes and dicentric chromosomes in metaphases (R>0.93, P 0.93, P<0.0001). The dose-response curves with gamma rays were very similar for NPB, ring chromosomes and dicentric chromosomes, as were the dose-responses for MNi, acentric rings and fragments. However, not all acentric chromosomes and dicentric chromosomes/centric rings were converted to MNi and NPB respectively, depending on the dose of radiation. Preliminary data, using FISH, suggests that NPB often represent DNA from a structural rearrangement involving only one or two homologous chromosomes. The results from this study validate the inclusion of NPB in the CBMN assay which provides a valuable measure of chromosome breakage/ rearrangement that was otherwise not available in the micronucleus assay. The CBMN assay allows NPB measurement to be achieved reliably because inhibition of cytokinesis prevents the loss of NPB that would otherwise occur if cells were allowed to divide

  13. p53 protein expression versus micronucleus induction as an indicator of DNA damage

    International Nuclear Information System (INIS)

    Hickman, A.W.; Carpenter, T.R.; Johnson, N.F.

    1994-01-01

    In vitro assays for detecting DNA damage play an important role in evaluating the possible adverse health effects of chemical compounds. Exposure to many DNA-damaging agents in vitro has been shown to cause elevated levels of the tumor-suppressor protein p53. Work in our laboratory has shown that induction of the p53 protein is useful as a biodosimeter for determining the radiation dose to cells. The purpose of this investigation was to compare the sensitivity of this assay to that of micronucleus induction, which is commonly used as a marker of radiation-induced damage

  14. Antioxidant evaluation of heterocyclic compounds by cytokinesis-block micronucleus assay.

    Science.gov (United States)

    Godevac, Dejan; Tesević, Vele; Vajs, Vlatka; Milosavljević, Slobodan; Stanković, Miroslava

    2013-03-01

    This article summarizes the results of using cytokinesis-block micronucleus (CBMN) assay to evaluate the antioxidant potential of heterocyclic compounds. Most studies were carried out with naturally occurring heterocyclic compounds such as plant polyphenols: flavonoids, xanthones, coumarins, and ellagitannins, or plant derived products (juices, extracts, supplements) rich in bioactive heterocyclic compounds. There are also some studies dealing with synthetic heterocyclic antioxidants. CBMN assay is an in vitro study that has been used to evaluate antioxidant and protective effects of heterocyclic compounds on induced chromosome aberration in human lymphocytes.

  15. Patient exposure and micronucleus assay during extracorporeal shock wave lithotripsy (ESWL) for kidney stones

    International Nuclear Information System (INIS)

    Shao, M.

    1992-01-01

    This study reports results of radiation exposure to 20 patients with kidney stones during Extracorporeal Shock Wave Lithotripsy (ESWL). A domestically made JT-ESWL-I Lithotripter was used. Data indicated that the amount of radiation exposure is related to the numbers, size, location and radiodensity of the stone, and also the number of shock wave and the time of fluoroscopy exposure given to the patients. The results of the micronucleus frequency of lymphocytes assay in the human peripheral blood are reported. This effect increased with increasing radiation exposure doses. (author). 4 refs., 2 tabs

  16. Micronucleus induction and reproductive death in a human cell line exposed to low-energy argon beam

    International Nuclear Information System (INIS)

    Courdi, A.; Mari, D.; Herault, J.; Chauvel, P.

    1995-01-01

    The aim of this study was to measure the biological efficiency of a low-energy argon beam (E=7.1 MeV/nucleon, LET=1590 keV/μm) on a human melanoma cell line (CAL4) established in our Institute. Two different methods were used: the micronucleus (MN) test and the colony-forming assay. MN are scored in binucleate cells (BNC) and are formed from acentric fragments or whole chromosomes that have not been incorporated into daughter nuclei at mitosis. The colony-forming assay quantifies reproductive death. Parallel experiments were run with cobalt gamma-rays for comparison. After Co irradiation, the MN-free BNC dose-response curve coincided with that of the loss of colony-forming ability, suggesting the potential of the former as a predictive test of cell killing. After Ar irradiation, there was a dissociation between the two effects, especially at high doses: cell death was greater than the frequency of BNC with MN. The inactivation cross-section was 74 μm 2 ; it was 39 μm 2 for MN yield. Therefore, the relative biological effectiveness (RBE) was higher for cell killing than for MN yield (0.8 and 0.5, respectively, at a Co dose of 3 Gy). The total MN count in BNC followed the same pattern of response as the fraction of BNC with MN. However, multiple (>2) MN in BNC were more frequently observed after low-dose Ar irradiation than after gamma-ray exposure (RBE > 1). Moreover, the frequency of multiple MN induction exceeded that expected from a Poisson distribution at all dose levels of Ar irradiation. (orig.)

  17. 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

  18. Tradescantia micronucleus test indicates genotoxic potential of traffic emissions in European cities

    International Nuclear Information System (INIS)

    Klumpp, Andreas; Ansel, Wolfgang; Klumpp, Gabriele; Calatayud, Vicent; Garrec, Jean Pierre; He Shang; Penuelas, Josep; Ribas, Angela; Ro-Poulsen, Helge; Rasmussen, Stine; Sanz, Maria Jose; Vergne, Phillippe

    2006-01-01

    Urban atmospheres contain complex mixtures of air pollutants including mutagenic and carcinogenic substances such as benzene, diesel soot, heavy metals and polycyclic aromatic hydrocarbons. In the frame of a European network for the assessment of air quality by the use of bioindicator plants, the Tradescantia micronucleus (Trad-MCN) test was applied to examine the genotoxicity of urban air pollution. Cuttings of Tradescantia clone no. 4430 were exposed to ambient air at 65 monitoring sites in 10 conurbations employing a standardised methodology. The tests revealed an elevated genotoxic potential mainly at those urban sites which were exposed to severe car traffic emissions. This bioassay proved to be a suitable tool to detect local 'hot spots' of mutagenic air pollution in urban areas. For its use in routine monitoring programmes, however, further standardisation of cultivation and exposure techniques is recommended in order to reduce the variability of results due to varying environmental conditions. - The Tradescantia micronucleus test can be used to assess genotoxic potential at urban sites

  19. Tradescantia micronucleus test indicates genotoxic potential of traffic emissions in European cities

    Energy Technology Data Exchange (ETDEWEB)

    Klumpp, Andreas [Institute for Landscape and Plant Ecology (320), University of Hohenheim, 70593 Stuttgart (Germany)]. E-mail: aklumpp@uni-hohenheim.de; Ansel, Wolfgang [Institute for Landscape and Plant Ecology (320), University of Hohenheim, 70593 Stuttgart (Germany); Klumpp, Gabriele [Institute for Landscape and Plant Ecology (320), University of Hohenheim, 70593 Stuttgart (Germany); Calatayud, Vicent [Fundacion CEAM, Parque Tecnologico, c/Charles Darwin 14, 46980 Paterna, Valencia (Spain); Garrec, Jean Pierre [INRA Nancy, Laboratoire Pollution Atmospherique, 54280 Champenoux (France); He Shang [INRA Nancy, Laboratoire Pollution Atmospherique, 54280 Champenoux (France); Penuelas, Josep [Unitat Ecofisiologia CSIC-CEAB-CREAF, Universitat Autonoma de Barcelona, Ed. C, 08193 Bellaterra, Barcelona (Spain); Ribas, Angela [Unitat Ecofisiologia CSIC-CEAB-CREAF, Universitat Autonoma de Barcelona, Ed. C, 08193 Bellaterra, Barcelona (Spain); Ro-Poulsen, Helge [Botanical Institute, University of Copenhagen, Oster Farimagsgade 2D, 1353 Copenhagen K (Denmark); Rasmussen, Stine [Botanical Institute, University of Copenhagen, Oster Farimagsgade 2D, 1353 Copenhagen K (Denmark); Sanz, Maria Jose [Fundacion CEAM, Parque Tecnologico, c/Charles Darwin 14, 46980 Paterna, Valencia (Spain); Vergne, Phillippe [ENS Lyon and Lyon Botanical Garden, 46 Allee d' Italie, 69364 Lyon Cedex 07 (France)

    2006-02-15

    Urban atmospheres contain complex mixtures of air pollutants including mutagenic and carcinogenic substances such as benzene, diesel soot, heavy metals and polycyclic aromatic hydrocarbons. In the frame of a European network for the assessment of air quality by the use of bioindicator plants, the Tradescantia micronucleus (Trad-MCN) test was applied to examine the genotoxicity of urban air pollution. Cuttings of Tradescantia clone no. 4430 were exposed to ambient air at 65 monitoring sites in 10 conurbations employing a standardised methodology. The tests revealed an elevated genotoxic potential mainly at those urban sites which were exposed to severe car traffic emissions. This bioassay proved to be a suitable tool to detect local 'hot spots' of mutagenic air pollution in urban areas. For its use in routine monitoring programmes, however, further standardisation of cultivation and exposure techniques is recommended in order to reduce the variability of results due to varying environmental conditions. - The Tradescantia micronucleus test can be used to assess genotoxic potential at urban sites.

  20. Examination of Mutagenic Effects of GAL-57 Herbicide (Bentazone+Dicamba Using Mouse Micronucleus Test

    Directory of Open Access Journals (Sweden)

    Vesela Karan

    2007-01-01

    Full Text Available A micronucleus test was run to investigate mutagenic potential of the herbicide GAL-57, a formulated mixture of bentazone and dicamba.The test was applied to mice of both sexes (strain: CRL: NMRI BR and the herbicide (product was administered by gavage at 2000 mg/kg rate, twice within 24 hs. Cyclophosphamide (positive control was administered at 60 mg/kg, while distilled water as a solvent was negative control. The animals were sacrificed 24 hs after second treatment, their bone marrow cells isolated from femur, and effects evaluated.The data acquired showed that repeated treatment of mice with GAL-57 caused neither biological nor significant statistical increase in the number of micronuclei in treated animals. At the same time, the number of micronucleated polychromatic erythrocytes in the bone marrow of animals treated with cyclophosphamide (positive control showed a significant statistical increase. The results suggest that the herbicide product tested did not show any mutagenic activity under the conditions of mouse micronucleus test.

  1. Comparison of micronucleus frequencies and proliferation kinetics in three X-irradiated cell lines

    International Nuclear Information System (INIS)

    Kaffenberger, W.; Becker, K.; Beuningen, D. van

    1990-01-01

    The kinetics of the occurrence of micronuclei was correlated with the survival of three mammalian cell lines of human, monkey, and mouse origin after irradiation with 240 kV X-rays. Particular attention was paid to the evaluation of the individual proliferation kinetics of the cell lines as well as to the characterization of micronuclei subpopulation with respect to size and possible biological importance using DNA and BUdR labelling techniques, fluorescence microscopy, and image analysis. The results demonstrate very characteristic size distributions of micronuclei for the three cell lines independent of radiation dose and time after irradiation. A close correlation between cell death and the occurrence of micronuclei (expressed as a calculated 'MN index') after irradiation could be established only when the kinetics of progression of cells through the cell cycle (e.g. the doubling time) and the biological characteristics of micronuclei (e.g. BUdR positivity, the micronucleus frequencies, and the number of micronuclei per main nucleus) were taken into account. Therefore, the micronucleus assay might not be useful as a quantitative perdictive assay in vivo but may allow qualitative estimations of radiation damage only because the necessary proliferation parameters of the cells might not be possible to establish in vivo. (orig.) [de

  2. Micronucleus assay as a biomarker of genotoxicity in the occupational exposure to agrochemicals in rural workers.

    Science.gov (United States)

    Gentile, N; Mañas, F; Bosch, B; Peralta, L; Gorla, N; Aiassa, D

    2012-06-01

    This paper aims to evaluate the genotoxic effect of agrochemicals in rural workers occupationally exposed by the micronucleus assay in peripheral blood lymphocytes and to promote the development of health and environmental preventive and protective practices. A total of 30 blood samples from 20 individuals occupationally exposed to different agrochemicals and 10 unexposed persons, who formed the reference group, were analyzed. We found statistically significant differences (p < 0.0005, Student's t Test) in the frequency of micronuclei between the two groups (7.20 ± 1.55 and 15.15 ± 5.10 CBMN for reference and exposed groups respectively). The analysis of age showed a positive correlation (Pearson Correlation Test) with the frequency of micronuclei in exposed population (p < 0.05; r(2) = 0.47), in contrast with smoking habits and years of exposure. Micronucleus assay allows an early detection of populations at higher risk of having genetic damage, allowing us to implement strategies of intervention for the purpose of contributing to reduce that risk.

  3. Hyperthermia-induced micronucleus formation in a human keratinocyte cell line

    International Nuclear Information System (INIS)

    Hintzsche, Henning; Riese, Thorsten; Stopper, Helga

    2012-01-01

    Elevated temperature can cause biological effects in vitro and in vivo. Many studies on effects of hypo- and hyperthermia have been conducted, but only few studies systematically investigated the formation of genomic damage in the micronucleus test in human cells in vitro as a consequence of different temperatures. In the present study, HaCaT human keratinocytes were exposed to different temperatures from 37 °C to 42 °C for 24 h in a regular cell culture incubator. Micronucleus frequency as a marker of genomic damage was elevated in a temperature-dependent and statistically significant manner. Apoptosis occurred at temperatures of 39 °C or higher. Cell proliferation was unaffected up to 40 °C and decreased at 41 °C and 42 °C. Expression of the heat shock protein Hsp70 was elevated, particularly at temperatures of 40 °C and higher. These findings are in agreement with several in vivo studies and some in vitro studies looking at single, specific temperatures, but a systematically investigated temperature-dependent increase of genomic damage in human keratinocytes in vitro is demonstrated for the first time here.

  4. Effects of estradiol and progesterone on the variability of the micronucleus assay

    International Nuclear Information System (INIS)

    Baeyens, Ans; Vandersickel, Veerle; Thierens, Hubert; Ridder, Leo De; Vral, Anne

    2005-01-01

    To investigate chromosomal radiosensitivity of lymphocytes the micronucleus (MN) assay has been used for many years. The results of these studies suggest the use of the MN assay as a biomarker for cancer predisposition. However, the MN assay has still some limitations associated with the reproducibility and sensitivity. Especially a high intra-individual variability has been observed. An explanation for this high intra-individual variability is not yet available. In literature it is suggested that the high variability among females is attributable to hormonal status. In this study we investigated if the high intra-individual variability in micronucleus formation in lymphocytes of females after in vitro exposure to ionising radiation is caused by variations in hormone levels of estradiol (E2) and progesterone (PROG). For this, the MN assay was performed on blood samples of 18 healthy women during 7 consecutive weeks while the estradiol and progesterone levels were determined at the same time. The MN assay was also examined in cultures of isolated blood lymphocytes with estradiol or progesterone levels added in vitro. The results demonstrated that estradiol and progesterone levels have no influence on the variations in radiation-induced MN yields observed in blood samples of healthy women. These conclusions were confirmed by the 'in vitro' experiments as no correlation between the MN yields and the concentrations of hormones (estradiol or progesterone) added in vitro to isolated lymphocytes cultures was observed

  5. Evaluation of a liver micronucleus assay in young rats (III): a study using nine hepatotoxicants by the Collaborative Study Group for the Micronucleus Test (CSGMT)/Japanese Environmental Mutagen Society (JEMS)-Mammalian Mutagenicity Study Group (MMS).

    Science.gov (United States)

    Takasawa, Hironao; Suzuki, Hiroshi; Ogawa, Izumi; Shimada, Yasushi; Kobayashi, Kazuo; Terashima, Yukari; Matsumoto, Hirotaka; Aruga, Chinami; Oshida, Keiyu; Ohta, Ryo; Imamura, Tadashi; Miyazaki, Atsushi; Kawabata, Masayoshi; Minowa, Shigenori; Hayashi, Makoto

    2010-04-30

    We have been investigating a liver micronucleus assay to detect genotoxic chemicals using young rats for several years, and had established its advantages with respect to using autonomous proliferation of young rat hepatocytes. Nine chemicals known to induce hepatotoxic effects such as necrosis (2,6-dinitrotolune, bromobenzene, isoniazid, phenacetin, allyl alcohol and thioacetamide), cholestasis (chlorpromazine hydrochloride and alpha-naphthyl isothiocyanate) and oxidative stress (clofibrate) were selected for this study. A liver micronucleus assay was conducted in 4-week-old male F344 rats using two or three dose levels of test chemicals given orally by gavage to evaluate the compound's ability to induce micronucleated hepatocytes. Several of these test chemicals were additionally examined in a peripheral blood micronucleus assay conducted concurrently and in the same animals. The genotoxic rodent hepatocarcinogen, 2,6-dinitrotoluene showed a positive result in the liver micronucleus assay, but the nongenotoxic hepatocarcinogens, clofibrate and thioacetamide gave negative responses. Bromobenzene, known to produce DNA adducts but is noncarcinogenic in rodent liver, was judged equivocal in this assay. alpha-Naphthyl isothiocyanate is noncarcinogenic and showed negative response in the liver. The other four chemicals, known to be either noncarcinogenic or carcinogenic in other non-liver target organs, showed negative results in the liver micronucleus assay. Based on the results in the present study and previous report described above, it was concluded that this technique is able to effectively predict genotoxic rodent hepatocarcinogenicity, and does not give false positives due to hepatotoxicity. Copyright 2010 Elsevier B.V. All rights reserved.

  6. Dependence of the bystander effect for micronucleus formation on dose of heavy-ion radiation in normal human fibroblasts

    International Nuclear Information System (INIS)

    Matsumoto, Yoshitaka; Hamada, Nobuyuki; Aoki-Nakano, Mizuho; Furusawa, Yoshiya; Funayama, Tomoo; Sakashita, Tetsuya; Kobayashi, Yasuhiko; Wada, Seiichi; Kakizaki, Takehiko

    2015-01-01

    Ionising radiation-induced bystander effects are well recognised, but its dependence on dose or linear energy transfer (LET) is still a matter of debate. To test this, 49 sites in confluent cultures of AG01522D normal human fibroblasts were targeted with microbeams of carbon (103 keV μm -1 ), neon (375 keV μm -1 ) and argon ions (1260 keV μm -1 ) and evaluated for the bystander-induced formation of micronucleus that is a kind of a chromosome aberration. Targeted exposure to neon and argon ions significantly increased the micronucleus frequency in bystander cells to the similar extent irrespective of the particle numbers per site of 1- 6. In contrast, the bystander micronucleus frequency increased with increasing the number of carbon-ion particles in a range between 1 and 3 particles per site and was similar in a range between 3 and 8 particles per site. These results suggest that the bystander effect of heavy ions for micronucleus formation depends on dose. (authors)

  7. Combined cytokinesis-block micronucleus and chromosomal aberration assay for the evaluation of radiosensitizers at low radiation doses

    International Nuclear Information System (INIS)

    Oya, Natsuo; Shibamoto, Yuta; Shibata, Toru

    1994-01-01

    Several methods have been tried for evaluating the efficacy of hypoxic cell radiosensitizers at clinically relevant low radiation doses (1-4 Gy). The cytokinesis-block micronucleus assay is known to be useful for both the in vitro and in vivo evaluation of radiosensitizers, while the chromosomal aberration assay has been commonly used to assess the mutagenicity of various agents. In the present study, the chromosomal aberration assay and the cytokinesis-block micronucleus assay were performed simultaneously to assess the radiosensitizing effect of etanidazole and KU-2285 at low radiation doses. The correlation between the two assays was also evaluated. In vitro study: EMT-6 cells were irradiated at a dose of 1-3 Gy under hypoxic conditions with or without the drugs at 1 mM. In vivo-in vitro study: EMT-6 tumor-bearing BALB/c mice received 2-4 Gy of radiation with or without administration of the drugs at 200 mg/kg. Single-cell suspensions were then obtained in both studies and were used for the cytokinesis-block micronucleus assay and the chromosomal aberration assay. The micronucleus frequency in binucleate cells was evaluated in the former assay, and the frequency of chromosomal aberrations in metaphase cells was evaluated in the latter assay. In vitro study: the sensitizer enhancement ratios of etanidazole and KU-2285 were 1.73 and 2.21, respectively, in the micronucleus assay, and 1.41 and 1.79 in the chromosomal aberration assay. In vivo-in vitro study: the sensitizer enhancement ratios of etanidazole and KU-2285 were 1.18 and 1.31, respectively, in the micronucleus assay, and 1.16 and 1.42 in the chromosomal aberration assay. In both studies, a linear correlation was observed between the micronucleus frequency and the chromosomal aberration frequency. The background (i.e., the frequency at 0 Gy) of the latter assay was considerably lower than that of the former assay, especially in the in vivo study. 31 refs., 4 figs

  8. Micronucleus frequency in Danish schoolchildren and their mothers from the DEMOCOPHES population

    DEFF Research Database (Denmark)

    Mørck, Thit A.; Vande Loock, Kim; Poulsen, Maria Bech

    2016-01-01

    organic pollutants and dioxin-like activity measured in the sameparticipants. The MN frequency analysis was performed with the cytokinesis-block micronucleus(CBMN) assay and included 100 children and 119 mothers. We found a significant correlationbetween mothers and children in the levels of micronuclei...... in 1000 binucleated T lymphocytes(‰MNBN) and in the proliferation index. Further the levels of ‰MNBN were significantly higherin mothers compared with their children. No significant associations were found for ‰MNBNfor traffic related exposure in neither children nor their mothers. In children, a 2.......5 times highermicronuclei in mononuclear T lymphocytes were found in children living within 50 m of a busyroad, however, this was not found in mothers or in MNBN and the effect of exposure to road trafficon MN frequency needs further investigation. No significant associations were found between...

  9. Micronucleus as biomarkers of cancer risk in anabolic androgenic steroids users.

    Science.gov (United States)

    Souza, L da Cunha Menezes; da Cruz, L A; Cerqueira, E de Moraes Marcílio; Meireles, Jrc

    2017-03-01

    The use of anabolic androgenic steroids (AAS) has grown among practitioners of recreational bodybuilding, with significant contributions of designer steroids, aiming muscle hypertrophy in healthy subjects. The abusive use of AAS in general is associated with adverse effects; one of the most worrisome is cancer development. The aim of this study was to evaluate the effectiveness of the cytokinesis block micronucleus (CBMN) test in human lymphocytes in identifying risk groups for cancer development in users of AAS. Blood was collected from 15 AAS users bodybuilders (G1), 20 non-users bodybuilders (G2) and 20 non-users sedentary (G3). MN analysis was performed on a minimum of 1000 binucleated lymphocytes. The occurrence of MN was significantly higher ( p < 0.05) in individuals of G1 compared to G2 and G3. The results indicate the sensitivity of CBMN in human lymphocytes in the identification of chromosomal damage in consequence of AAS.

  10. Significance of the proportion of binucleate cells in the micronucleus assay; A methodological study

    Energy Technology Data Exchange (ETDEWEB)

    Imamura, Masahiro; Edgren, M.R. (Karolinska Inst., Stockholm (Sweden). Dept. of Radiation Physics)

    1994-03-01

    Using treatment with cytochalasin-B (Cyt-B) for the induction of a cytokinetic block, the significance of the proportion of binucleate cells (BNC) in the micronucleus (MN) assay was investigated in a methodological study. A Chinese hamster cell line V79 was used in which MN were induced by radiation. In complementary tests the radiation effect in inducing MN was enhanced by depletion of the cellular glutathione content with buthionine sulfoximine (BSO). The data indicated that the concentration of Cyt-B is the major factor which determines the proportion of BNC. This proportion was shown to be independent of radiation dose and of BSO. Furthermore, the MN frequency was not related to the percentage of BNC. Therefore, a high proportion of BNC may be practical for the MN assay, but may not make the technique more accurate. (author).

  11. Significance of the proportion of binucleate cells in the micronucleus assay

    International Nuclear Information System (INIS)

    Imamura, Masahiro; Edgren, M.R.

    1994-01-01

    Using treatment with cytochalasin-B (Cyt-B) for the induction of a cytokinetic block, the significance of the proportion of binucleate cells (BNC) in the micronucleus (MN) assay was investigated in a methodological study. A Chinese hamster cell line V79 was used in which MN were induced by radiation. In complementary tests the radiation effect in inducing MN was enhanced by depletion of the cellular glutathione content with buthionine sulfoximine (BSO). The data indicated that the concentration of Cyt-B is the major factor which determines the proportion of BNC. This proportion was shown to be independent of radiation dose and of BSO. Furthermore, the MN frequency was not related to the percentage of BNC. Therefore, a high proportion of BNC may be practical for the MN assay, but may not make the technique more accurate. (author)

  12. Genotoxicity and ELF-magnetic fields: a review through the micronucleus assay

    International Nuclear Information System (INIS)

    Alcaraz, M.; Andreu-Galvez, M.; Sanchez-Villalobos, J. M.; Achel, D. G.; Olmos, E.; Martinez-Hernandez, C. M.

    2012-01-01

    Thirty for (34) published studies, conducted from 1994 to the present to evaluate the genotoxic effect of magnetic fields using ELF-EMF and diagnostic resonance on humans by the micronucleus assay have been reviewed. some characteristics of the assay methods, their significance to genotoxicity and basic interpretations of the results of these assays are discussed. of the studies analysed 70.5% implicated genotoxic effects induced by these magnetic fields: 52.9% were due to exposure to magnetic fields only and 17,6% by exposure to magnetic fields in combination with some treatment types, resulting in additive or synergistic effect. Evidence exist to support the notion that exposure of humans to magnetic fields stimulates genotoxic effects, although the actual mechanisms of action or even the true human health consequences resulting from these exposure still remain unclear. (Author) 80 refs.

  13. Genotoxicity assessment of cobalt chloride in Eisenia hortensis earthworms coelomocytes by comet assay and micronucleus test.

    Science.gov (United States)

    Ciğerci, İbrahim Hakkı; Ali, Muhammad Muddassir; Kaygısız, Şöhret Yüksek; Liman, Recep

    2016-02-01

    Cobalt and its different compounds are extensively used worldwide and considered as possible environmental pollutant. Earthworms are useful model organism and its different species are used to monitor soil pollution. No study has been found to detect cobalt chloride (CoCl2) genotoxicity in earthworms. So, current study aimed to evaluate CoCl2 induced genotoxicity in Eisenia hortensis earthworms coelomocytes by alkaline comet assay (CA) and micronucleus (MN) test. The earthworms (n = 10 for each group) were exposed to different series of CoCl2 concentrations (100 ppm, 200 ppm, 300 ppm, 400 ppm, 500 ppm, 600 ppm) to find LD50. The LD50 for CoCl2 was found at 226 ppm. Then, doses of LD50/2, LD50 and 2XLD50 for 48 h were used. CA and MN demonstrated the significant increase (P earthworms. Copyright © 2015 Elsevier Ltd. All rights reserved.

  14. Structural and numerical chromosome aberration inducers in liver micronucleus test in rats with partial hepatectomy.

    Science.gov (United States)

    Itoh, Satoru; Hattori, Chiharu; Nagata, Mayumi; Sanbuissho, Atsushi

    2012-08-30

    The liver micronucleus test is an important method to detect pro-mutagens such as active metabolites not reaching bone marrow due to their short lifespan. We have already reported that dosing of the test compound after partial hepatectomy (PH) is essential to detect genotoxicity of numerical chromosome aberration inducers in mice [Mutat. Res. 632 (2007) 89-98]. In naive animals, the proportion of binucleated cells in rats is less than half of that in mice, which suggests a species difference in the response to chromosome aberration inducers. In the present study, we investigated the responses to structural and numerical chromosome aberration inducers in the rat liver micronucleus test. Two structural chromosome aberretion inducers (diethylnitrosamine and 1,2-dimethylhydrazine) and two numerical chromosome aberration inducers (colchicine and carbendazim) were used in the present study. PH was performed a day before or after the dosing of the test compound in 8-week old male F344 rats and hepatocytes were isolated 4 days after the PH. As a result, diethylnitrosamine and 1,2-dimethylhydrazine, structural chromosome aberration inducers, exhibited significant increase in the incidence of micronucleated hepatocyte (MNH) when given either before and after PH. Colchicine and carbendazim, numerical chromosome aberration inducers, did not result in any toxicologically significant increase in MNH frequency when given before PH, while they exhibited MNH induction when given after PH. It is confirmed that dosing after PH is essential in order to detect genotoxicity of numerical chromosome aberration inducers in rats as well as in mice. Regarding the species difference, a different temporal response to colchicine was identified. Colchicine increased the incidence of MNH 4 days after PH in rats, although such induction in mice was observed 8-10 days after PH. Copyright © 2012 Elsevier B.V. All rights reserved.

  15. Analysis of the Genotoxic Effects of Mobile Phone Radiation using Buccal Micronucleus Assay: A Comparative Evaluation.

    Science.gov (United States)

    Banerjee, Sumita; Singh, Narendra Nath; Sreedhar, Gadiputi; Mukherjee, Saikat

    2016-03-01

    Micronucleus (MN) is considered to be a reliable marker for genotoxic damage and it determines the presence and the extent of the chromosomal damage. The MN is formed due to DNA damage or chromosomal disarrangements. The MN has a close association with cancer incidences. In the new era, mobile phones are constantly gaining popularity specifically in the young generation, but this device uses radiofrequency radiation that may have a possible carcinogenic effect. The available reports related to the carcinogenic effect of mobile radiation on oral mucosa are contradictory. To explore the effects of mobile phone radiation on the MN frequency in oral mucosal cells. The subjects were divided into two major groups: low mobile phone users and high mobile phone users. Subjects who used their mobile phone since less than five years and less than three hours a week comprised of the first group and those who used their mobile since more than five years and more than 10 hours a week comprised of the second group. Net surfing and text messaging was not considered in this study. Exfoliated buccal mucosal cells were collected from both the groups and the cells were stained with DNA-specific stain acridine orange. Thousand exfoliated buccal mucosal cells were screened and the cells which were positive for micronuclei were counted. The micronucleus frequency was represented as mean±SD, and unpaired Student t-test was used for intergroup comparisons. The number of micronucleated cells/ 1000 exfoliated buccal mucosal cells was found to be significantly increased in high mobile phone users group than the low mobile phone users group. The use of mobile phone with the associated complaint of warmth around the ear showed a maximum increase in the number of micronucleated cells /1000 exfoliated buccal mucosal cells. Mobile phone radiation even in the permissible range when used for longer duration causes significant genotoxicity. The genotoxicity can be avoided to some extent by the

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

    Directory of Open Access Journals (Sweden)

    D Cavallo

    2010-01-01

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

  17. Use of LysoTracker dyes: a flow cytometric study of autophagy.

    Science.gov (United States)

    Chikte, Shaheen; Panchal, Neelam; Warnes, Gary

    2014-02-01

    The flow cytometric use of LysoTracker dyes was employed to investigate the autophagic process and to compare this with the upregulation of autophagy marker, the microtubule-associated protein LC3B. Although the mechanism of action of LysoTracker dyes is not fully understood, they have been used in microscopy to image acidic spherical organelles, and their use in flow cytometry has not been thoroughly investigated in the study of autophagy. This investigation uses numerous autophagy-inducing agents including chloroquine (CQ), rapamycin, low serum (used to analyze patient cells as well as easier to use and significantly less costly. Copyright © 2013 International Society for Advancement of Cytometry.

  18. 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...

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

    International Nuclear Information System (INIS)

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

    1996-01-01

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

  20. 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.

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

    Science.gov (United States)

    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.

  2. 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

  3. 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.

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

    Science.gov (United States)

    Józwa, Wojciech; Czaczyk, Katarzyna

    2012-04-02

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

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

    International Nuclear Information System (INIS)

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

    2006-01-01

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

  6. 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.

  7. [Influence of Four Kinds of PPCPs on Micronucleus Rate of the Root-Tip Cells of Vicia-faba and Garlic].

    Science.gov (United States)

    Wang, Lan-jun; Wang, Jin-hua; Zhu, Lu-sheng; Wang, Jun; Zhao, Xiang

    2016-04-15

    In order to determine the degree of biological genetic injury induced by PPCPs, the genotoxic effects of the doxycycline (DOX), ciprofloxacin (CIP), triclocarban (TCC) and carbamazepine (CBZ) in the concentration range of 12.5-100 mg · L⁻¹ were studied using micronucleus rate and micronucleus index of Vicia-fabe and garlic. The results showed that: (1) When the Vicia-faba root- tip cells were exposed to DOX, CIP, TCC and CBZ, micronucleus rates were higher than 1.67 ‰ (CK₁), it was significantly different from that of the control group (P garlic root tip cells were exposed to DOX, CIP, TCC and CBZ respectively, the micronucleus rates were less than those of the Vicia-faba, while in most treatments significantly higher than that of the control group (0.67‰). The micronucleus index was higher than 3.5 in the groups exposed to CIP with concentrations of 25, 50, 100 mg · L⁻¹ and TCC and CBZ with concentrations of 25 mg · L⁻¹; With the increase of exposure concentrations, the micronucleus rate showed a trend of first increasing and then decreasing as well. (3) Under the same experimental conditions, the cells micronucleus rates of the garlic cells caused by the four tested compounds were significantly lower than those of Vicia-faba. (4) The micronucleus index of the root tip cells of Vicia-faba and garlic treated with the four kinds of compounds followed the order of CIP > CBZ > TCC > DOX. These results demonstrated that the four compounds caused biological genetic injury to root-tip cells of Vicia-faba and garlic, and the genetic damage caused to garlic was significantly lower than that to Vicia-faba. The damages caused by the four kinds of different compounds were also different.

  8. Spontaneous micronucleus frequencies in human peripheral blood lymphocytes as a screening test for an individual variation in a different population and radiation-induced micronucleus induction

    International Nuclear Information System (INIS)

    Kang, Chang-Mo; Jeon, Hye-Jeong; Cho, Chul-Koo

    2004-01-01

    Our studies were to evaluate the role of epigenetic factors in the variation of radiosensitivity on human peripheral blood lymphocytes by measuring the frequencies of micronucleus (MN) from 293 healthy subjects of different population for assessing the radiation health risk in Korea. We analyzed the frequencies of both spontaneous and in vitro 60 Co γ-rays or 50MeV neutron-induced MNs. The frequencies of spontaneous NMs not only vary greatly between individuals, but also working or living areas. The increased levels of cells with spontaneous MNs were observed with an increasing age. The frequencies of spontaneous MNs were significantly higher in females than in males. For both sexes, MN frequency was significantly and positively correlated with age. Age and gender are the most important demographic variables impacting on the MN index. Donors who had ever smoked showed significantly increased frequencies of MNs compared to nonsmokers. The main lifestyle factors influencing the MN index in the subjects are correlated significantly and positively with smoke while measuring the spontaneous frequencies of micronuclei. Therefore, it is evident that with regard to the application of MN assay all future studies to evaluate the association between radiosensitivity and susceptibility for radiation health risks in different populations should take into account the effect of age, gender and lifestyle. For the dose-response study, the induced MNs were observed at all doses, and the numerical changes according to doses. The dose-response curves were fitted with a linear-quadratic forms of the dose, and the results were different for γ-rays and neutrons significantly. Neutrons were more effective than γ-rays in producing MN with a dose-dependent manner. The frequency of MN varies with dose. The RBE for a micronuclei was 2.37 ± 0.17. The results suggested that the MN assay have a high potential to ensure appropriate quality control and a standard documentation protocol, which

  9. Evaluation of genetic alteration induced by radon gas using the micronucleus test (Tradescantia sp. clone KU-20)

    International Nuclear Information System (INIS)

    Bruschi, Armando L.; Azevedo, Heliana de; Macacini, Jose F.; Roque, Claudio V.

    2011-01-01

    The first observations over the existence of radon gas (Rn), initially known as 'thorium emanation', were carried out between the end of 19 th and beginning of 20 th centuries. A result of uranium-238 (U 238 ) radioactive decay, radon is a tasteless, odorless and colorless gas under room temperature, with a 3.825-day half life and particle α emission in its decay, and as final product of its disintegration, the stable lead-206 isotope (Pb 206 ). Being it is the gas with the highest density known, closed and poor ventilated environments are favorable to its accumulation, with its inhalation being the highest health risk. The use of vegetal bioindicators has shown to be excellent on the monitoring of air quality and on mutagenic potential of various pollutants contained in the atmosphere. Within this context, the objective of this study was to evaluate the micronucleus test application potential utilizing the Tradescantia sp. clone KU-20, in order to evaluate genetic alterations induced by radon gas. Stems of Tradescantia sp. clone KU-20, previously immerse in Hoagland solution, were introduced in a radon detection equipment's calibration chamber (Alphaguard), containing radium salt. Afterwards, the accommodated stems were exposed to radon gas (the average radon concentration was 7.639 KBq/m3) for 24 hours. The results demonstrated an increase on micronucleus formation (39.23 + 2.143 MCN/100 tetrads) in stems exposed in relation to the negative control (18.00 + 1.396 MCN/100 tetrads). The difference between the values indicated a significant increase on micronucleus frequency in the inflorescences subjected to radon gas. The presented results demonstrated the micronucleus test application potential using Tradescantia clone KU-20 to evaluate genetic effects induced by radon gas. (author)

  10. Cytokinesis-block micronucleus assay evolves into a 'cytome' assay of chromosomal instability, mitotic dysfunction and cell death

    International Nuclear Information System (INIS)

    Fenech, Michael

    2006-01-01

    The cytokinesis-block micronucleus (CBMN) assay was originally developed as an ideal system for measuring micronuclei (MNi) however it can also be used to measure nucleoplasmic bridges (NPBs), nuclear buds (NBUDs), cell death (necrosis or apoptosis) and nuclear division rate. Current evidence suggests that (a) NPBs originate from dicentric chromosomes in which the centromeres have been pulled to the opposite poles of the cell at anaphase and are therefore indicative of DNA mis-repair, chromosome rearrangement or telomere end-fusions, (b) NPBs may break to form MNi, (c) the nuclear budding process is the mechanism by which cells remove amplified and/or excess DNA and is therefore a marker of gene amplification and/or altered gene dosage, (d) cell cycle checkpoint defects result in micronucleus formation and (e) hypomethylation of DNA, induced nutritionally or by inhibition of DNA methyl transferase can lead to micronucleus formation either via chromosome loss or chromosome breakage. The strong correlation between micronucleus formation, nuclear budding and NPBs (r = 0.75-0.77, P < 0.001) induced by either folic acid deficiency or exposure to ionising radiation is supportive of the hypothesis that folic acid deficiency and/or ionising radiation cause genomic instability and gene amplification by the initiation of breakage-fusion-bridge cycles. In its comprehensive mode, the CBMN assay measures all cells including necrotic and apoptotic cells as well as number of nuclei per cell to provide a measure of cytotoxicity and mitotic activity. The CBMN assay has in fact evolved into a 'cytome' method for measuring comprehensively chromosomal instability phenotype and altered cellular viability caused by genetic defects and/or nutrional deficiencies and/or exogenous genotoxins thus opening up an exciting future for the use of this methodology in the emerging fields of nutrigenomics and toxicogenomics and their combinations

  11. Assessment of the genotoxicity of Cu and Zn in raw and anaerobically digested slurry with the Vicia faba micronucleus test

    OpenAIRE

    Marcato, Claire-Emmanuelle; Pinelli, Eric; Pourrut, Bertrand; Silvestre, Jérôme; Guiresse, Agnès Maritchù

    2009-01-01

    Genotoxicity of Cu and Zn was assessed by use of the micronucleus (MN) test on Vicia faba roots. Plants were exposed to various leachates of rawand anaerobically digested pig slurry, with maximum total concentrations of 200MCu and 600MZn. The results indicated stabilisation of the organic matter during anaerobic digestion of the slurry and bioconversion of some phytotoxic organic compounds (e.g. phenols or p-cresol), but did not showa relationship between Cu and Zn concentrations and MN fr...

  12. In vivo micronucleus test as a biomarker of genotoxicity in free-range goats from suspected contaminated environment

    Directory of Open Access Journals (Sweden)

    Afusat Jagun Jubril

    2017-09-01

    Conclusion: The finding indicates the prevalence and frequency of micronucleus as a biomarker of genotoxicity and an indicator of exposure to environmental genotoxic subtances. Hence, this highlights the relevance of these goats as important sentinel animal model. These findings, therefore, serve as a preliminary data for further studies on the latent genotoxic environmental contaminants and their potential deleterious impact. [J Adv Vet Anim Res 2017; 4(3.000: 281-287

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

    International Nuclear Information System (INIS)

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

    2012-01-01

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

  14. Comet assay and micronucleus tests on Oreochromis niloticus (Perciforme: Cichlidae) exposed to raw sugarcane vinasse and to phisicochemical treated vinasse by pH adjustment with lime (CaO).

    Science.gov (United States)

    Correia, Jorge E; Christofoletti, Cintya Ap; Ansoar-Rodríguez, Yadira; Guedes, Thays A; Fontanetti, Carmem S

    2017-04-01

    In Brazil vinasse, a main sugarcane distillery residue, stands out because every liter of alcohol generates 10-15 L of vinasse as waste. An alternative for the disposal of this waste is the fertirrigation of the sugarcane culture itself. However, the high amount released can saturate the soil and through leaching/percolation contaminate water resources. The aim of this study is verifying the toxic potential of vinasse in tilapias and effectiveness of the physicalchemical treatment of this waste with pH adjustment with lime (CaO). The comet assay and the micronucleus test were applied on animals exposed to dilutions of raw vinasse and vinasse adjusted to neutral pH. Bioassays with raw vinasse dilutions indicated a toxic and genotoxic potential; fish exposed to the highest concentration died less than 48 h after the exposure; the incidence of micronucleus was significantly higher when compared to negative control for all dilutions. For the comet assay, the scores of damage were statistically higher for all dilutions, with the exception of the 1% dillution. However, in the bioassay with the chemically treated vinasse (neutral pH), most fish in the 10% dilution survived and there was no significant difference when compared to the control. Damage scores in the comet assay were similar to the results of the untreated vinasse. The chemical treatment of vinasse with lime to neutralize the pH proved to be an effective alternative for the toxicity reduction of this residue, since it reduced the mortality of fish at higher concentrations and the incidence of damage to DNA. Copyright © 2017 Elsevier Ltd. All rights reserved.

  15. 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.

  16. 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 ...

  17. 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

  18. Kinetics of micronucleus induction and cytotoxicity caused by distinct antineoplastics and alkylating agents in vivo.

    Science.gov (United States)

    Morales-Ramírez, Pedro; Vallarino-Kelly, Teresita; Cruz-Vallejo, Virginia

    2014-01-30

    This mini-review aims to compare the differences in the kinetics of the induction of micronucleated polychromatic erythrocytes (MN-PCE) and cytotoxicity by distinct antineoplastic and genotoxic agents in murine peripheral blood in vivo and to correlate these kinetics with the underlying processes. Comparisons were carried out using our previously obtained data with nominal doses causing similar levels of cytotoxicity, as measured in terms reduction of PCE. The aneuploidogens caused the most rapid induction of MN-PCEs and had the highest rates of cytotoxicity and genotoxicity. The promutagens cyclophosphamide and dimethylnitrosamine showed the most delayed responses and had the lowest genotoxic and cytotoxic efficiencies. DNA crosslinking agents had a similar delay of 4-5 h, greater than those of aneuploidogens, but differed in their cytotoxic and genotoxic efficiencies. Methylnitrosourea and 5-aza-cytidine caused greater delays than crosslinking agents. These delays can be due to the methylnitrosourea-mediated induction of formation of mono alkyl adducts which are interpreted as mismatches during DNA duplication, whereas 5-aza-cytidine requires incorporation into the DNA to induce breakage. This review allows us to conclude that the requirement for metabolic activation and the mechanisms of DNA breakage and of micronucleus induction are the main factors that affect the time of maximal MN-PCE induction. Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.

  19. Inhibitory effects of Enteromorpha linza polysaccharide on micronucleus of Allium sativum root cells.

    Science.gov (United States)

    Zhang, Zhongshan; Wang, Xiaomei; Li, Jingfen; Liu, Chongbin; Zhang, Quanbin

    2016-06-01

    In this study, the antimutagenic function of the polysaccharide from Enteromorpha linza with the micronucleus test of Allium sativum root cells induced by sulfur dioxide and ultraviolet was studied. The concentration-effect relation of the two inducers was firstly evaluated. The results showed that an increase of genotoxicity damage was demonstrated and micronuclei frequency induced by sulfur dioxide and ultraviolet displayed dose dependent increases. All the doses of polysaccharide did affect the micronuclei frequency formation compared with the negative control. And also, the significant increase in inhibition rate of micronuclei frequency was observed with the increase of the dose of polysaccharide. It was showed maximum inhibition of micronuclei frequency cells (71.74% and 66.70%) at a concentration of 200g/mL in three experiments. The low molecular weight polysaccharide showed higher inhibition rate than raw polysaccharide at the higher concentration (50g/mL) in the absence of sulfur dioxide and ultraviolet. It was confirmed to be a good mutant inhibitor. Copyright © 2016 Elsevier B.V. All rights reserved.

  20. Effect of mobile phone station on micronucleus frequency and chromosomal aberrations in human blood cells.

    Science.gov (United States)

    Yildirim, M S; Yildirim, A; Zamani, A G; Okudan, N

    2010-01-01

    The use of mobile telephones has rapidly increased worldwide as well as the number of mobile phone base stations that lead to rise low level radiofrequency emissions which may in turn have possible harm for human health. The national radiation protection board has published the known effects of radio waves exposure on humans living close to mobile phone base stations. However, several studies have claimed that the base station has detrimental effects on different tissues. In this study, we aimed to evaluate the effects of mobile phone base stations on the micronucleus (MN) frequency and chromosomal aberrations on blood in people who were living around mobile phone base stations and healthy controls. Frequency of MN and chromosomal aberrations in study and control groups was 8.96 +/- 3.51 and 6.97 +/- 1.52 (p: 0.16); 0.36 +/- 0.31 and 0.75 +/- 0.61 (p: 0.07), respectively. Our results show that there was not a significant difference of MN frequency and chromosomal aberrations between the two study groups. The results claim that cellular phones and their base stations do not produce important carcinogenic changes.

  1. Use of micronucleus test in the assessment of radiation effects in aquatic environments

    International Nuclear Information System (INIS)

    Araujo, Edvaldo F. de; Silva, Luanna R.S.; Lima, Pedro A. de S.; Amancio, Francisco F.; Melo, Ana Maria M. de A.; Silva, Edvane B. da; Silva, Ronaldo C. da

    2011-01-01

    The study of the effects of radioactive substances on the environment is accomplished by radioecology. This science has played an important role in combating all forms of pollution. The uncontrolled use of physical and chemical agents has been a concern for environmental regulatory agencies, due to the serious damage to ecosystems. Aquatic organisms are exposed to a variety of pollutants harmful to aquatic systems. The mollusks Biomphalaria glabrata has been featured as a bioindicator to possess characteristics such as short reproductive cycle ease of maintenance in the laboratory and low maintenance cost. The micronucleus assay has been shown to be a great test to identify mutagenic effects caused by physical and chemical agents. In this study the frequency of micronuclei in haemocytes of Biomphalaria glabrata exposed to high doses of 60 Co gamma radiation contributing to a further standardization of this test as an indicator of the presence of radioactive contamination in aquatic environments. The young adult snails of Biomphalaria glabrata were divided into groups and subjected to a dose of 0 (control), 40 and 60 Gy of gamma radiation. The results showed that snails irradiated with 40 Gy showed a smaller number of haemocytes, whereas those exposed to 60 Gy had a greater quantity of these cells compared to control group. It can be concluded that the morphological analysis and the frequency of micronuclei in haemocytes of Biomphalaria glabrata exposed to 60 Co gamma radiation may be used in studies of the action of high doses of radiation in aquatic environments (author)

  2. Effects of heavy metals/metalloids contamination of soils on micronucleus induction in Tradescantia pallida

    Directory of Open Access Journals (Sweden)

    Neelima Meravi

    2013-06-01

    Full Text Available The present study was conducted in GGV campus, Bilaspur in which heavy metals/metalloids speciation of soil (for Cr, Fe, Ni, Cd and Pb was performed for assessing the genotoxicity of these metals. The metals concentrations were measured with the help of AAS 7000 (Shimadzu and the standard solution was prepared using standard metal solution of Inorganic Ventures. The concentrations of Cr, Fe, Ni, Cd and Pb (in ug/100 g soil were 12.4, 33.9, 3.1, 0.07 and 2.4 respectively. The flowers of Tradescantia pallida plants growing in this soil were taken and their micronucleus (Trad-MCN bioassay was performed. Trad-MCN bioassay was performed using the protocols established by Ma (1981. The study revealed that at these concentrations of metals micronuclei (stained objects that were smaller than the nuclei and not connected to the nuclei are classified as MCN were formed. Therefore it can be inferred from the present study that soil of GGV campus is genotoxic for the Tradescantia pallida.

  3. Micronucleus Assay in Exfoliated Buccal Epithelial Cells Using Liquid Based Cytology Preparations in Building Construction Workers.

    Science.gov (United States)

    Arul, P; Smitha, Shetty; Masilamani, Suresh; Akshatha, C

    2018-01-01

    Cytogenetic damage in exfoliated buccal epithelial cells due to environmental and occupational exposure is often monitored by micronucleus (MN) assay using liquid based cytology (LBC) preparations. This study was performed to evaluate MN in exfoliated buccal epithelial cells of building construction workers using LBC preparations. LBC preparations of exfoliated buccal epithelial cells from 100 subjects [50 building construction workers (cases) and 50 administrative staffs (controls)] was evaluated by May-Grunwald Giemsa, Hematoxylin and Eosin and Papanicolaou stains. Student's t test was used for statistical analysis and a P value of 5 years) and smokers and non-smokers of cases (P=0.001). However, there were meaningful differences regarding mean frequencies of MN between smokers, non-smokers, those with alcohol consumption or not in cases and controls using various stains (P=0.001). There was an increased risk of cytogenetic damage in building construction workers. However, evaluation of MN of exfoliated buccal epithelial cells in building construction workers serve as a minimally invasive biomarker for cytogenetic damage. LBC preparations can be applied for MN assay as it improves the quality of smears and cell morphology, decreases the confounding factors and reduces false positive results.

  4. Synergistic interaction of radiation and octylphenol evaluated by tradescantia-micronucleus assay

    International Nuclear Information System (INIS)

    Shin, H. S.; Lee, J. H.; Lee, B. H.; Kim, J. K.

    2003-01-01

    Many kinds of synthetic chemicals have been being used for various purposes. Some of them are called 'endocrine disruptors' because they can disturb the endocrine system of organisms. Presently no technique is established for the quantitative assessment of biological risk of the environmental hormones. The pollen mother cells (PMC) of tradescantia are very sensitive to chemical toxicants or ionizing radiation, and thus can be used as a biological end- point for assessing their effects. Micronucleus frequencies in PMC showed a good dose- and concentration-response relationship for radiation and bisphenol A. The MCN frequencies in the pollen mother cells treated with octylphenol were 4.20, 7.27, 4.93 MCN/100 tetrads for 1, 5 and 10 μM, respectively. On the other hand, the frequencies were 10.13, 19.27, 24.47 MCN/100 tetrads for the octylphenol treatments (1, 5, and 10 μM) combined with 30 cGy irradiation. The MCN frequency of 30 cGy control was 8.00 MCN/100 tetrads for the octylphenol treatments (1, 5, and 10μM) combined with 30 cGy irradiation. The MCN frequency of 30 cGy control was 8.00 MCN/100 tetrads. It is known from the result that the Trad-MCN assay can be an excellent tool for the detection of biologically harmful effects of environmental toxicants or synthetic chemicals

  5. Biomonitoring of Serbian population revealed by CB micronucleus test before and after the bombing of Yugoslavia

    International Nuclear Information System (INIS)

    Joksic, G.; Stankovic, M.; Guc-Scekic, M.; Vranjes, A.

    2002-01-01

    The induction of micronuclei in mitotically active cells has been widely used and promoted as a biological marker of exposure to environmental toxins. Biomonitoring of population using cytochalasin block micronucleus test (CBMN) has been performed for 11 years in our country; the incidence of micronuclei was evaluated in many groups of occupationally exposed persons as well as healthy unexposed controls. The spontaneous frequency of MN per 1000 binucleated cells was 9±3 (mean±SD) for woman, 7±2 for men. The average incidence of micronuclei in lymphocytes of newborns was 5.3±0.6, in their mothers 15±3 per 1000 binucleated cells, respectively. After the bombing of Yugoslavia significantly higher incidence of micronuclei was found in all groups of examines: health adults and newborns. In healthy adults, the average incidence of micronuclei was 28.16±14.63; in young pregnant woman 25.3±5.02 and their foetuses 20.14±9.6 respectively. One year latter (2001) the incidence of MN declined in all adults but enhance in foetal blood lymphocytes. (author)

  6. Nuclear localization signal targeting to macronucleus and micronucleus in binucleated ciliate Tetrahymena thermophila.

    Science.gov (United States)

    Iwamoto, Masaaki; Mori, Chie; Osakada, Hiroko; Koujin, Takako; Hiraoka, Yasushi; Haraguchi, Tokuko

    2018-06-08

    Ciliated protozoa possess two morphologically and functionally distinct nuclei: a macronucleus (MAC) and a micronucleus (MIC). The MAC is transcriptionally active and functions in all cellular events. The MIC is transcriptionally inactive during cell growth, but functions in meiotic events to produce progeny nuclei. Thus, these two nuclei must be distinguished by the nuclear proteins required for their distinct functions during cellular events such as cell proliferation and meiosis. To understand the mechanism of the nuclear transport specific to either MAC or MIC, we identified specific nuclear localization signals (NLSs) in two MAC- and MIC-specific nuclear proteins, macronuclear histone H1 and micronuclear linker histone-like protein (Mlh1), respectively. By expressing GFP-fused fragments of these proteins in Tetrahymena thermophila cells, two distinct regions in macronuclear histone H1 protein were assigned as independent MAC-specific NLSs and two distinct regions in Mlh1 protein were assigned as independent MIC-specific NLSs. These NLSs contain several essential lysine residues responsible for the MAC- and MIC-specific nuclear transport, but neither contains any consensus sequence with known monopartite or bipartite NLSs in other model organisms. Our findings contribute to understanding how specific nuclear targeting is achieved to perform distinct nuclear functions in binucleated ciliates. © 2018 The Authors. Genes to Cells published by Molecular Biology Society of Japan and John Wiley & Sons Australia, Ltd.

  7. Fluorescence in situ hybridization in combination with the comet assay and micronucleus test in genetic toxicology

    Directory of Open Access Journals (Sweden)

    Hovhannisyan Galina G

    2010-09-01

    Full Text Available Abstract Comet assay and micronucleus (MN test are widely applied in genotoxicity testing and biomonitoring. While comet assay permits to measure direct DNA-strand breaking capacity of a tested agent MN test allows estimating the induced amount of chromosome and/or genome mutations. The potential of these two methods can be enhanced by the combination with fluorescence in situ hybridization (FISH techniques. FISH plus comet assay allows the recognition of targets of DNA damage and repairing directly. FISH combined with MN test is able to characterize the occurrence of different chromosomes in MN and to identify potential chromosomal targets of mutagenic substances. Thus, combination of FISH with the comet assay or MN test proved to be promising techniques for evaluation of the distribution of DNA and chromosome damage in the entire genome of individual cells. FISH technique also permits to study comet and MN formation, necessary for correct application of these methods. This paper reviews the relevant literature on advantages and limitations of Comet-FISH and MN-FISH assays application in genetic toxicology.

  8. 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

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

    Directory of Open Access Journals (Sweden)

    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.

  10. 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

  11. 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

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

    Directory of Open Access Journals (Sweden)

    Serkan Yazıcı

    2015-01-01

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

  13. Multivariate analysis of flow cytometric data using decision trees

    Directory of Open Access Journals (Sweden)

    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.

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

    Science.gov (United States)

    Jenkins, J A; Draugelis-Dale, R O; Pinkney, A E; Iwanowicz, L R; Blazer, V S

    2015-03-15

    Declining harvests of yellow perch, Perca flavescens, in urbanized watersheds of Chesapeake Bay have prompted investigations of their reproductive fitness. The purpose of this study was to establish a flow cytometric technique for DNA analysis of fixed samples sent from the field to provide reliable gamete quality measurements. Similar to the sperm chromatin structure assay, measures were made on the susceptibility of nuclear DNA to acid-induced denaturation, but used fixed rather than live or thawed cells. Nuclei were best exposed to the acid treatment for 1 minute at 37 °C followed by the addition of cold (4 °C) propidium iodide staining solution before flow cytometry. The rationale for protocol development is presented graphically through cytograms. Field results collected in 2008 and 2009 revealed DNA fragmentation up to 14.5%. In 2008, DNA fragmentation from the more urbanized watersheds was significantly greater than from reference sites (P = 0.026) and in 2009, higher percentages of haploid testicular cells were noted from the less urbanized watersheds (P = 0.032) indicating better reproductive condition at sites with less urbanization. For both years, total and progressive live sperm motilities by computer-assisted sperm motion analysis ranged from 19.1% to 76.5%, being significantly higher at the less urbanized sites (P < 0.05). This flow cytometric method takes advantage of the propensity of fragmented DNA to be denatured under standard conditions, or 1 minute at 37 °C with 10% buffered formalin-fixed cells. The study of fixed sperm makes possible the restrospective investigation of germplasm fragmentation, spermatogenic ploidy patterns, and chromatin compaction levels from samples translocated over distance and time. The protocol provides an approach that can be modified for other species across taxa. Published by Elsevier Inc.

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

    Directory of Open Access Journals (Sweden)

    Bence György

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

  16. Study design and statistical analysis of data in human population studies with the micronucleus assay.

    Science.gov (United States)

    Ceppi, Marcello; Gallo, Fabio; Bonassi, Stefano

    2011-01-01

    The most common study design performed in population studies based on the micronucleus (MN) assay, is the cross-sectional study, which is largely performed to evaluate the DNA damaging effects of exposure to genotoxic agents in the workplace, in the environment, as well as from diet or lifestyle factors. Sample size is still a critical issue in the design of MN studies since most recent studies considering gene-environment interaction, often require a sample size of several hundred subjects, which is in many cases difficult to achieve. The control of confounding is another major threat to the validity of causal inference. The most popular confounders considered in population studies using MN are age, gender and smoking habit. Extensive attention is given to the assessment of effect modification, given the increasing inclusion of biomarkers of genetic susceptibility in the study design. Selected issues concerning the statistical treatment of data have been addressed in this mini-review, starting from data description, which is a critical step of statistical analysis, since it allows to detect possible errors in the dataset to be analysed and to check the validity of assumptions required for more complex analyses. Basic issues dealing with statistical analysis of biomarkers are extensively evaluated, including methods to explore the dose-response relationship among two continuous variables and inferential analysis. A critical approach to the use of parametric and non-parametric methods is presented, before addressing the issue of most suitable multivariate models to fit MN data. In the last decade, the quality of statistical analysis of MN data has certainly evolved, although even nowadays only a small number of studies apply the Poisson model, which is the most suitable method for the analysis of MN data.

  17. Dose-response relationship of octylphenol and radiation evaluated by tradescantia-micronucleus assay

    International Nuclear Information System (INIS)

    Kim, J. K.; Cheon, K. J.; Lee, B. H.; Shin, H. S.; Lee, J. H.

    2002-01-01

    Many kinds of synthetic chemicals have been being used for various purposes. Some of them are called 'Endocrine Disruptor's because they can disturb the endocrine system of organisms. Presently no technique is established for the quantitative assessment of biological risk of the environmental hormones. The pollen mother cells (PMC) of Tradescantia are very sensitive to chemical toxicants or ionizing radiation, and thus can be used as a biological end-point assessing their effect. Micronucleus frequencies in PMC showed a good dose- and concentration-response relationship for radiation, bisphenol A and octylphenol. A parallel series of experiment using five increasing doses of gamma-ray at 10, 20, 30, 40 and 50 cGy was conducted. The MCN frequencies of 12.0, 25.2, 41.7, 76 and 83 MCN/100 tetrads were observed from each of the increasing gamma-ray dosage groups, respectively. Lenear regression analysis of the gamma-ray data MCN frequencies yielded a correlation coefficient of 0.95. the MCN frequencies in pollen mother cells treated with bisphenol a and octylphenol showed dose-response relationship in a concentration of 0, 1, 2, 4 μM and 0, 4, 10, 20 μM. the MCN frequency for the bisphenol a and octylphenol group yields 2.33, 8.06, 12.7 and 19.6 MCN/100 tetrads for the bisphenol a and 2.33, 2.33, 11.47, 17.6 MCN/100 tetrads for the octylphenol. The MCN frequency of the control was 2.33 MCN/100 tetrads. It is known from the result that Trad-MCN assay can be an excellent tool for detection of biological risk due to environmental toxicants or synthetic chemicals

  18. Micronucleus evaluation for determining the chromosomal breakages after radionuclide synovectomy in patients with hemophilia

    International Nuclear Information System (INIS)

    Kavakli, K.; Cogulu, O.; Karaca, E.

    2012-01-01

    To investigate the genotoxic effects of 90 Y and 186 Re in patients with hemophilia who were undergoing radionuclide synovectomy (RS) procedure in the last 3 years. Nineteen patients were enrolled in the study. Most of the patients (n=17) were hemophilia-A (mean age 20.6±10.5 years) and 18 patients (mean age 22.6±10.6 years) with hemophilia who were not exposed to RS procedure were included in the study as control group. Most cases in the control group (n=13) were hemophilia-A. 90 Y for knee joints and 186 Re for elbow or ankle joints were used to perform RS in hemophilic patients. We studied the micronucleus (MN) test on peripheral blood lymphocytes as an indicator of radiation-induced cytogenetic damage and calculated nuclear division index. There was no significant difference between the patients with and without RS with respect to MN values. However, both values obtained in RS-exposed patients and control group were much elevated than values reported in literature from healthy controls. The mean MN values of patients below 20 years old were much lower but not significant than those above 20 years old. MN frequencies between 186 Re and 90 Y groups were also analyzed, and no significant difference was observed. Hemophilia patients who were treated with 186 Re showed higher levels of MN compared to patients treated with 90 Y although the difference was not significant. Radioisotope synovectomy (RS) seems to be a safe procedure not causing a significant genotoxic effect on hemophilic patients, however, further studies including larger series of patients are needed to better understand the effects of RS on patients' health. (author)

  19. Scoring of radiation-induced micronuclei in cytokinesis-blocked human lymphocytes by automated image analysis

    International Nuclear Information System (INIS)

    Verhaegen, F.; Seuntjens, J.; Thierens, H.

    1994-01-01

    The micronucleus assay in human lymphocytes is, at present, frequently used to assess chromosomal damage caused by ionizing radiation or mutagens. Manual scoring of micronuclei (MN) by trained personnel is very time-consuming, tiring work, and the results depend on subjective interpretation of scoring criteria. More objective scoring can be accomplished only if the test can be automated. Furthermore, an automated system allows scoring of large numbers of cells, thereby increasing the statistical significance of the results. This is of special importance for screening programs for low doses of chromosome-damaging agents. In this paper, the first results of our effort to automate the micronucleus assay with an image-analysis system are represented. The method we used is described in detail, and the results are compared to those of other groups. Our system is able to detect 88% of the binucleated lymphocytes on the slides. The procedure consists of a fully automated localization of binucleated cells and counting of the MN within these cells, followed by a simple and fast manual operation in which the false positives are removed. Preliminary measurements for blood samples irradiated with a dose of 1 Gy X-rays indicate that the automated system can find 89% ± 12% of the micronuclei within the binucleated cells compared to a manual screening. 18 refs., 8 figs., 1 tab

  20. 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.

  1. 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

  2. Micronucleus test of varying amounts of potassium bromate (KBrO3) on the meristematic cells of Allium cepa var. aggregatum root tips

    International Nuclear Information System (INIS)

    Cajigal Romnick, M.; Somera, Leomerto A.

    1999-03-01

    Four hundred twenty onion bulbs of the multiplier variety Allium cepa var. aggregatum were used as test materials to assay the micronucleus induction capacity of potassium bromate doses of 0, 5, 10, 25, 50, 75, and 100 parts per million. Microscopic analyses were done using onion root tips prepared according to a modified technique of Medina (1994). These analyses were done on root tips taken from onions grown in KBrO 3 for three days and for five days. The study was conducted following a completely randomized design and the data were statistically analyzed using a non-parametric equivalent of the analysis of variance. A significant amount of micronucleated cells (MCN) were found among treated onions compared with the almost non-occurrence in the control groups (0 ppm). The Kruskal-Wallis H-test and the Wilcoxon two-samples tests revealed significant differences among treatment means and that a significant increase in the number of MCN occurs as the dose of KBr0 3 increased in both day experiments. Results from the higher doses of 50, 75, and 100 ppm were found to be significantly the same for the day 3 experiments while those of the day 5 higher doses are characterized by lack of clear cellular and nuclear outline such that scoring is difficult. Differences in MCN averages for the day 3 and 5 experiments appear to be insignificant. However, day 3 results show averages that are more significantly different from each other. These prove that the MCN can be used as an efficient and time-saving parameter for the allium test of chemicals with chromosome breaking capacities. (Author)

  3. Micronucleus test of varying amounts of potassium bromate (KBrO{sub 3}) on the meristematic cells of Allium cepa var. aggregatum root tips

    Energy Technology Data Exchange (ETDEWEB)

    Cajigal Romnick, M; Somera, Leomerto A

    1999-03-01

    Four hundred twenty onion bulbs of the multiplier variety Allium cepa var. aggregatum were used as test materials to assay the micronucleus induction capacity of potassium bromate doses of 0, 5, 10, 25, 50, 75, and 100 parts per million. Microscopic analyses were done using onion root tips prepared according to a modified technique of Medina (1994). These analyses were done on root tips taken from onions grown in KBrO{sub 3} for three days and for five days. The study was conducted following a completely randomized design and the data were statistically analyzed using a non-parametric equivalent of the analysis of variance. A significant amount of micronucleated cells (MCN) were found among treated onions compared with the almost non-occurrence in the control groups (0 ppm). The Kruskal-Wallis H-test and the Wilcoxon two-samples tests revealed significant differences among treatment means and that a significant increase in the number of MCN occurs as the dose of KBr0{sub 3} increased in both day experiments. Results from the higher doses of 50, 75, and 100 ppm were found to be significantly the same for the day 3 experiments while those of the day 5 higher doses are characterized by lack of clear cellular and nuclear outline such that scoring is difficult. Differences in MCN averages for the day 3 and 5 experiments appear to be insignificant. However, day 3 results show averages that are more significantly different from each other. These prove that the MCN can be used as an efficient and time-saving parameter for the allium test of chemicals with chromosome breaking capacities. (Author)

  4. 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).

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

    Directory of Open Access Journals (Sweden)

    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.

  6. 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

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

    Directory of Open Access Journals (Sweden)

    Gentry-Nielsen Martha J

    2006-07-01

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

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

    Directory of Open Access Journals (Sweden)

    Coignoul Freddy

    2007-09-01

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

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

    Science.gov (United States)

    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.

  10. Alkaline Comet Assay and Micronucleus Test Parameters in Children Exposed to Diagnostic X-Ray Examination

    International Nuclear Information System (INIS)

    Gajski, G.; Geric, M.; Garaj-Vrhovac, V.; Milkovic, Dj.; Beck, N.; Ranogajec-Komor, M.; Miljanic, S.; Knezevic, Z.

    2011-01-01

    Chest radiograms represent the basic radiological examination of thorax and are the most frequently performed radiological diagnostic procedure in the child population. Understanding the risks of low doses of radiation is an important aspect in the risk benefit analysis in paediatric populations. To provide the best care for the young patients the effects of radiation should be minimized thus chest X-rays must be performed by highest standards to ensure that the young patient has the lowest risk possible. Since children are the most sensitive to radiation, there is a need for follow up of the young populations that receive these X-ray diagnostic examinations. Follow up would be especially advisable for children that are at higher risk of radiation induced damage, for example children with a predisposition to DNA damage, or for children that are constantly exposed to numerous radiological examinations due to their illness. In that manner, present study was undertaken to evaluate application of different dosimetry systems in conjunction with alkaline comet assay and micronucleus test for the assessment of different types of DNA and chromosomal alterations in child population exposed to acute diagnostic X-rays examination. For that purpose doses were measured using thermoluminescence (TL) and radiophotoluminescent (RPL) dosimetry systems. The study demonstrated that immediately after exposure to diagnostic X-irradiation, mean percentage of DNA in tail of the comets, which is indirect measures of DNA damage, was significantly changed. The same was noticed for mean total number of micronuclei as well. It was shown that children with pulmonary diseases subjected to diagnostic procedure develop a significant increase in mean total number of each measured parameter which are the biomarkers of genetic damage for carcinogenesis, than prior to diagnostic procedure and that interindividual differences exist for each monitored child. Our results show that genetic damage arises

  11. Genotoxic potential generated by biomass burning in the Brazilian Legal Amazon by Tradescantia micronucleus bioassay: a toxicity assessment study

    Directory of Open Access Journals (Sweden)

    Artaxo Paulo

    2011-05-01

    Full Text Available Abstract Background The Brazilian Amazon has suffered impacts from non-sustainable economic development, especially owing to the expansion of agricultural commodities into forest areas. The Tangará da Serra region, located in the southern of the Legal Amazon, is characterized by non-mechanized sugar cane production. In addition, it lies on the dispersion path of the pollution plume generated by biomass burning. The aim of this study was to assess the genotoxic potential of the atmosphere in the Tangará da Serra region, using Tradescantia pallida as in situ bioindicator. Methods The study was conducted during the dry and rainy seasons, where the plants were exposed to two types of exposure, active and passive. Results The results showed that in all the sampling seasons, irrespective of exposure type, there was an increase in micronucleus frequency, compared to control and that it was statistically significant in the dry season. A strong and significant relationship was also observed between the increase in micronucleus incidence and the rise in fine particulate matter, and hospital morbidity from respiratory diseases in children. Conclusions Based on the results, we demonstrated that pollutants generated by biomass burning in the Brazilian Amazon can induce genetic damage in test plants that was more prominent during dry season, and correlated with the level of particulates and elevated respiratory morbidity.

  12. Biomonitoring of genotoxic risk in radar facility workers: comparison of the comet assay with micronucleus assay and chromatid breakage assay

    International Nuclear Information System (INIS)

    Garaj-Vrhovac, V.; Kopjar, N.

    2003-01-01

    Genotoxic risks of occupational exposure in a radar facility were evaluated by using alkaline comet assay, micronucleus assay and chromatid breakage assay on peripheral blood leukocytes in exposed subjects and corresponding controls. Results show that occupational exposure to microwave radiation correlates with an increase of genome damage in somatic cells. The levels of DNA damage in exposed subjects determined by using alkaline comet assay were increased compared to control and showed interindividual variations. Incidence of micronuclei was also significantly increased compared to baseline control values. After short exposure of cultured lymphocytes to bleomycin, cells of occupationally exposed subjects responded with high numbers of chromatid breaks. Although the level of chromosome damage generated by bleomycin varied greatly between individuals, in exposed subjects a significantly elevated number of chromatid breaks was observed. Our results support data reported in literature indicating that microwave radiation represents a potential DNA-damaging hazard. Alkaline comet assay is confirmed as a sensitive and highly reproducible technique for detection of primary DNA damage inflicted in somatic cells. Micronucleus assay was confirmed as reliable bio-markers of effect and chromatid breakage assay as sensitive bio-marker of individual cancer susceptibility. The results obtained also confirm the necessity to improve measures and to perform accurate health surveillance of individuals occupationally exposed to microwave radiation

  13. 3-Nitrobenzanthrone (3-NBA) induced micronucleus formation and DNA damage in human hepatoma (HepG2) cells.

    Science.gov (United States)

    Lamy, Evelyn; Kassie, Fekadu; Gminski, Richard; Schmeiser, Heinz H; Mersch-Sundermann, Volker

    2004-01-15

    3-Nitrobenzanthrone (3-NBA), identified in diesel exhaust and in airborne particulate matter, is a potent mutagen in Salmonella, induces micronuclei formation in mice and in human cells and DNA adducts in rats. In the present study, we investigated the genotoxic potency of 3-NBA in human HepG2 cells using the micronucleus (MN) assay and the single cell gel electrophoresis (SCGE). 3-NBA caused a genotoxic effect at concentrations > or =12 nM in both assays. In the micronucleus assay, we found 98.7+/-10.3 MN/1000 BNC at a concentration of 100 nM 3-NBA in comparison to 27.3+/-0.6 MN/1000 BNC with the negative control. At the same concentration, the DNA-migration (SCGE) showed an Olive tail moment (OTM) of 2.7+/-0.45 and %DNA in the tail of 8.28+/-0.76; OTM and %DNA in the tail of cells treated with the negative control were 0.73+/-0.08 and 2.81+/-0.30, respectively. The results are discussed under consideration of former studies.

  14. Biomonitoring of agricultural workers exposed to pesticide mixtures in Guerrero state, Mexico, with comet assay and micronucleus test.

    Science.gov (United States)

    Carbajal-López, Yolanda; Gómez-Arroyo, Sandra; Villalobos-Pietrini, Rafael; Calderón-Segura, María Elena; Martínez-Arroyo, Amparo

    2016-02-01

    The aim of this study was to evaluate the genotoxic effect of pesticides in exfoliated buccal cells of workers occupationally exposed in Guerrero, Mexico, using the comet assay and the micronucleus test. The study compared 111 agricultural workers in three rural communities (Arcelia 62, Ajuchitlan 13, and Tlapehuala 36), with 60 non-exposed individuals. All the participants were males. The presence of DNA damage was investigated in the exfoliated buccal cells of study participants with the comet assay and the micronucleus (MN) test; comet tail length was evaluated in 100 nuclei and 3000 epithelial cells of each individual, respectively; other nuclear anomalies such as nuclear buds, karyolysis, karyorrhexis, and binucleate cells were also evaluated. Study results revealed that the tail migration of DNA and the frequency of MN increased significantly in the exposed group, which also showed nuclear anomalies associated with cytotoxic or genotoxic effect. No positive correlation was noted between exposure time and tail length and micronuclei frequencies. No significant effect on genetic damage was observed as a result of age, smoking, and alcohol consumption. The MN and comet assay in exfoliated buccal cells are useful and minimally invasive methods for monitoring genetic damage in individuals exposed to pesticides. This study provided valuable data for establishing the possible risk to human health associated with pesticide exposure.

  15. Allegheny County Walk Scores

    Data.gov (United States)

    Allegheny County / City of Pittsburgh / Western PA Regional Data Center — Walk Score measures the walkability of any address using a patented system developed by the Walk Score company. For each 2010 Census Tract centroid, Walk Score...

  16. 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.

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

    Science.gov (United States)

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

    2017-10-23

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

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

    Directory of Open Access Journals (Sweden)

    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.

  19. 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

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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.

  2. 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.

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

    Science.gov (United States)

    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.

  4. 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.

  5. 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.

  6. Advantages of micronuclei analysis through images autocapturing and screen scoring

    International Nuclear Information System (INIS)

    González, J.E.; Martínez-López, W.

    2015-01-01

    The cytokinesis-block micronucleus (CBMN) test is a quantitative assay for genetic toxicity assessment. One of the advantages of the MN assay is its amenability for automation. Different type of cells has been used to evaluate genetic damage through MN assay, such as, human lymphocytes and rodent cell lines (i.e. CHO, V79, CHL and L5178Y). The MN quantification is a time consuming process and several efforts has been conducted for its automation. Some of them include an operator checking step, like PathFinder CellScan System, or are fully automated such as MNScore from MetaSytems. Usually, fully automated systems detect two or three times less MN than visual scoring. In some cases, the impact of false positive detection is reduced with a visual detection step. In the present work we have tested a combination of image autocapturing of CHOK1 cells previously treated with bleomycin (0, 2.5, 5.0 and 10.0 μg/ml) or UVC (0, 4, 8 and 16 J/m”2 ) with a screen scoring. Capturing images using the AutoCapture option from Metafer 4 from MetaSystems (GmbH, Germany) plus screen scoring render similar results in terms of MN cells frequency than microscopic live scoring. The resultant bias from the Bland–Altman analysis was -1.1% with confidence intervals between -2.2% and -0.1%, indicating an acceptable agreement between both MN scoring method. However, the mean time devoted to live microscope scoring per sample was 159 minutes compared to 39 minutes for microscope images autocapturing and screen scoring. Therefore, it become advantageous to combine autocapturing of microscope images plus screen scoring when many samples have to be analyzed for radiological biodosimetry purposes. (authors)

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

    Directory of Open Access Journals (Sweden)

    Fernando Fernández-Bañares

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

  8. 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

  9. Application of liquid-based cytology preparation in micronucleus assay of exfoliated buccal epithelial cells in road construction workers.

    Science.gov (United States)

    Arul, P

    2017-01-01

    Asphalts are bitumens that consist of complex of hydrocarbon mixtures and it is used mainly in road construction and maintenance. This study was undertaken to evaluate the micronucleus (MN) assay of exfoliated buccal epithelial cells in road construction workers using liquid-based cytology (LBC) preparation. Three different stains (May-Grunwald Giemsa, hematoxylin and eosin, and Papanicolaou) were used to evaluate the frequency of MN in exfoliated buccal epithelial of 100 participants (fifty road construction workers and fifty administrative staff) using LBC preparation. Statistical analysis was performed with Student's t-test, and Proad construction exhibit a higher frequency of MN in exfoliated buccal epithelial cells and they are under the significant risk of cytogenetic damage. LBC preparation has potential application for the evaluation of frequency of MN. This technique may be advocated in those who are occupationally exposed to potentially carcinogenic agents in view of improvement in the smear quality and visualization of cell morphology.

  10. Micronucleus formation compared to the survival rate of human melanoma cells after X-ray and neutron irradiation and hyperthermia

    Energy Technology Data Exchange (ETDEWEB)

    van Beuningen, D.; Streffer, C.; Bertholdt, G.

    1981-09-01

    After neutron and X-ray irradiation and combined X-ray irradiation and hyperthermia (3 hours, 42/sup 0/C), the survival rate of human melanoma cells was measured by means of the colony formation test and compared to the formation of micronuclei. Neutrons had a stronger effect on the formation of micronuclei than the combination of X-rays and hyperthermia. X-rays had the lowest effect. The dose effect curve showed a break at that dose level at which a reduction of cells was observed in the cultures. A good relation between survival rate and formation of micronuclei was found for the X-ray irradiation, but not for the neutron irradiation and the combined treatment. These observations are discussed. At least for X-rays, the micronucleus test has turned out to be a good screening method for the radiosensitivity of a biologic system.

  11. Genotoxic, Cytotoxic, Antigenotoxic, and Anticytotoxic Effects of Sulfonamide Chalcone Using the Ames Test and the Mouse Bone Marrow Micronucleus Test.

    Directory of Open Access Journals (Sweden)

    Carolina Ribeiro E Silva

    Full Text Available Chalcones present several biological activities and sulfonamide chalcone derivatives have shown important biological applications, including antitumor activity. In this study, genotoxic, cytotoxic, antigenotoxic, and anticytotoxic activities of the sulfonamide chalcone N-{4-[3-(4-nitrophenylprop-2-enoyl]phenyl} benzenesulfonamide (CPN were assessed using the Salmonella typhimurium reverse mutation test (Ames test and the mouse bone marrow micronucleus test. The results showed that CPN caused a small increase in the number of histidine revertant colonies in S. typhimurium strains TA98 and TA100, but not statistically significant (p > 0.05. The antimutagenicity test showed that CPN significantly decreased the number of His+ revertants in strain TA98 at all doses tested (p 0.05. Additionally, CPN co-administered with MMC significantly increased PCE/NCE ratio at all doses tested, demonstrating its anticytotoxic effect. In summary, CPN presented genotoxic, cytotoxic, antigenotoxic, and anticytotoxic properties.

  12. Evaluation of genotoxicity after application of Listerine(R) on human lymphocytes by micronucleus and single cell gel electrophoresis assays.

    Science.gov (United States)

    Türkez, Hasan; Togar, Basak; Arabaci, Taner

    2012-04-01

    Listerine (LN) is one of the most commonly used mouth rinses worldwide although very limited information is available concerning its genotoxicity. In another view, the biological safety profile of oral care products is frequently assumed on the basis of simplistic test models. Therefore, the present study was undertaken to investigate the in vitro genotoxic potential of LN using micronucleus and single cell gel electrophoresis tests as genetic endpoints. Different concentrations of LN (0-100% of ml/culture, v/v) were applied to whole human blood cultures (n = 5). The result of the present study showed that there were no statistically significant differences (p > 0.05) between the control group and the groups treated with LN alone in both analysed endpoints. In conclusion, our result first demonstrated the absence of genotoxicity of LN on human lymphocytes.

  13. 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

  14. 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)

  15. Bio-monitoring for the genotoxic assessment in road construction workers as determined by the buccal micronucleus cytome assay.

    Science.gov (United States)

    Çelik, Ayla; Yildirim, Seda; Ekinci, Seda Yaprak; Taşdelen, Bahar

    2013-06-01

    Buccal micronucleus cytome (BMCyt) assay monitors genetic damage, cell proliferation and cell death in humans exposed to occupational and environmental agents. BMCyt is used as an indicator of genotoxic exposure, since it is associated with chromosomal instability. There is little research on the occupational exposure among road construction workers for genotoxicity testing. In the present study, we evaluated MN frequencies and other nuclear changes, karyorrhexis (KR), karyolysis (KL), broken egg (BE), binucleate (BN), condensed chromatin cell (CCC), and picnotic cell (PC) in buccal mucosa cells of 40 road construction workers (twenty smokers and twenty non-smokers) and 40 control groups consisting of healthy persons (twenty smokers and twenty non-smokers). Microscopic observation was performed of 2000 cells per individual in both road construction workers and control group. In control and worker groups, for each person repair index (RI) was calculated via formula KR+L/BE+MN. The results showed a statistically significant increase in the frequency of MN in buccal epithelial cells of exposed group compared with control group (proad construction workers, RI is lower than the control group. There is a significant difference between workers and control group (proad paving operations are absorbed by workers and that asphalt fume exposure is able to significantly induce cytogenetic damage in buccal mucosa cells of workers after controlling some possible confounding factors, such as age, sex and smoking habits. In addition to determination of nuclear changes and the micronucleus, the determination of RI value presents a new approach to genotoxic bio-monitoring assessment studies of occupationally exposed population. Copyright © 2013 Elsevier Inc. All rights reserved.

  16. Fish eco-genotoxicology: Comet and micronucleus assay in fish erythrocytes as in situ biomarker of freshwater pollution

    Directory of Open Access Journals (Sweden)

    Bilal Hussain

    2018-02-01

    Full Text Available Owing to white meat production Labeo rohita have vast economic importance, but its population has been reduced drastically in River Chenab due to pollution. Atomic absorption spectrophotometry showed a merciless toxicity level of Cd, Cu, Mn, Zn, Pb, Cr, Sn and Hg. Comet assay results indicated significant (p < .05 DNA fragmentation in Labeo rohita as 42.21 ± 2.06%, 31.26 ± 2.41% and 21.84 ± 2.21% DNA in comet tail, tail moment as 17.71 ± 1.79, 10.30 ± 1.78 and 7.81 ± 1.56, olive moment as 13.58 ± 1.306, 8.10 ± 1.04 and 5.88 ± 0.06, respectively, from three different polluted sites on the river. Micronucleus assay showed similar findings of single micronucleus induction (MN as 50.00 ± 6.30‰, double MN 14.40 ± 2.56‰, while nuclear abnormalities (NA were found as 150.00 ± 2.92‰. These higher frequencies of MN induction and NA were found to be the cause of reduction of 96% of the population of this fish species in an experimental area of the River Chenab. This fish species has been found near extinction through the length of the river Chenab and few specimens in rainy seasons if restored by flood, may die in sugarcane mill season. Due to sweeping extinction Labeo rohita showed the highest sensitivity for pollution and could be used as bioindicator and DNA fragmentation in this column feeder fish species as a biomarker of the pollution load in freshwater bodies.

  17. Sacha Inchi (Plukenetia volubilis L. powder: acute toxicity, 90 days oral toxicity study and micronucleus assay in rodents

    Directory of Open Access Journals (Sweden)

    Idania Rodeiro

    2018-02-01

    Full Text Available Context: Sacha Inchi has been consumed for years by indigenous peoples. Meanwhile, its toxicological potential has not been sufficiently studied. Aims: To assess the acute, sub-chronic toxicity and genotoxicity evaluation of Sacha Inchi powder obtained from Plukenetia volubilis L. Methods: A dose of 2000 mg/kg was orally administered to rats and mice and toxicity symptoms for 14 days were observed. In repeated dose study, the product was orally administered to Sprague Dawley rats of both sexes. Animals received 50, 250 and 500 mg/kg/day of the product for 90 days. At the end, animals were sacrificed and samples were done for hematological and biochemical analysis, organ weighs and histopathological examination. Genotoxicity potential of Sacha Inchi powder was evaluated through micronucleus test in mice. Negative controls received the vehicle (carboxymethyl cellulose, 0.5% used. Results: No morbidity or mortality at 2000 mg/kg of the product were found. Sacha Inchi powder oral administration during 90 days to rats did not lead to death, body weight gain, food consumption, or adverse events. No significant changes on hematological or biochemical parameters, organ weights or histopathological findings were observed. Induction of micronucleus formation attributable to the product was not found in mice. Conclusions: No toxicity effects after oral acute exposure of Sacha Inchi power to rats and mice were observed. Neither toxicity attributable to oral doses of the product up to 500 mg/kg during 90 days to rats were found. Results suggested Sacha Inchi powder does not have genotoxicity potential under our experimental conditions.

  18. 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.

  19. 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.

  20. 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

  1. 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.

  2. 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

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

    Science.gov (United States)

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

    2011-01-01

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

  4. The Zhongshan Score

    Science.gov (United States)

    Zhou, Lin; Guo, Jianming; Wang, Hang; Wang, Guomin

    2015-01-01

    Abstract In the zero ischemia era of nephron-sparing surgery (NSS), a new anatomic classification system (ACS) is needed to adjust to these new surgical techniques. We devised a novel and simple ACS, and compared it with the RENAL and PADUA scores to predict the risk of NSS outcomes. We retrospectively evaluated 789 patients who underwent NSS with available imaging between January 2007 and July 2014. Demographic and clinical data were assessed. The Zhongshan (ZS) score consisted of three parameters. RENAL, PADUA, and ZS scores are divided into three groups, that is, high, moderate, and low scores. For operative time (OT), significant differences were seen between any two groups of ZS score and PADUA score (all P RENAL showed no significant difference between moderate and high complexity in OT, WIT, estimated blood loss, and increase in SCr. Compared with patients with a low score of ZS, those with a high or moderate score had 8.1-fold or 3.3-fold higher risk of surgical complications, respectively (all P RENAL score, patients with a high or moderate score had 5.7-fold or 1.9-fold higher risk of surgical complications, respectively (all P RENAL and PADUA scores. ZS score could be used to reflect the surgical complexity and predict the risk of surgical complications in patients undergoing NSS. PMID:25654399

  5. Changes of chromosome aberration rate and micronucleus frequency along with accumulated dose in continuously irradiated mice with a low dose rate of γ-rays

    International Nuclear Information System (INIS)

    Tanaka, Kimio; Izumi, Jun; Yanai, Takanori; Ichinohe, Kazuaki; Matsumoto, Tsuneya

    2003-01-01

    Chromosome aberrations in chronically exposed workers in nuclear facilities and medical radiologists have been reported. However chronological change of chromosome aberration rates along with accumulated dose has not been well studied. Chromosome aberrations and micronuclei in spleen lymphocytes were observed serially in mice continuously irradiated with a low dose rate of 20 mGy/day up to 400 days. Chromosome aberration rates were rapidly increased to 11.1% at 1 Gy, while micronucleus incidence increased at 5 Gy. After these doses their increase rates were saturated. Micronucleus incidence in bone marrow erythroblasts was higher than in spleen cells. These chronological changes of cytogenetic aberrations seem to be induced through a balance between developments of chromosome aberrations and micronuclei, and life span of spleen lymphocytes. These results will be helpful for risk assessment in low dose rate radiation exposure. (author)

  6. Use of the micronucleus assay for the selective detection of radiosensitivity in BUdR-unincorporated cells after pulse-labelling of exponentially growing tumour cells

    Energy Technology Data Exchange (ETDEWEB)

    Masunaga, S.; Ono, K.; Fushiki, M.; Abe, M. (Kyoto Univ. (Japan). Faculty of Medicine); Wandl, E.O. (Vienna Univ. (Austria). Klinik fuer Strahlentherapie und Strahlenbiologie)

    1990-08-01

    To determine the radiosensitivity of non S-phase tumour cells in vitro, survival curves of SCC VII tumour cells were obtained after a short block with hydroxyurea. Dose-response curves of micronucleus (MN) frequency appearing in non-S-phase cells were also determined by excluding S-phase cells with immunofluorescence staining to 5-bromo-2'-deoxyuridine (BUdR). Both the dose response curves of MN frequency and survival curves were analysed by a linear-quadratic model (surviving fraction =exp (-{alpha}D-{beta}D{sup 2}), MN frequency =aD+bD{sup 2}+c). A good correlation between the {alpha}/{beta} and a/b ratios was observed. In both BUdR-unincorporated and asynchronous cell cultures, the regression lines between the surviving fraction and micronucleus frequency were statistically identical. (author).

  7. Genotoxicity of freshwater ecosystem shows DNA damage in preponderant fish as validated by in vivo micronucleus induction in gill and kidney erythrocytes.

    Science.gov (United States)

    Obiakor, M O; Okonkwo, J C; Ezeonyejiaku, C D

    2014-12-01

    Genotoxicity of Anambra River was studied by micronucleus (MN) assay of preponderant fish species in the river. The micronucleus indices obtained were used as biomarker to estimate and predict pollution profile and possible danger of feeding on the aquatic species. Micronuclei profile of the fish was measured from gill and kidney erythrocytes using microscopic technique. Season, species and location effects on micronuclei, together with their interactions were also determined. Two major seasons (rainy and dry) and preponderant fish species in the river (Synodontis clarias, Linnaeus, 1758 and Tilapia nilotica, Linnaeus, 1757) were studied at five distinct locations that displayed differential environmental stresses. The study showed that the micronucleus index of fish is an excellent biomarker for measuring pollution level and genotoxicity of freshwater habitat. Season, species of fish and location affect micronuclei profile of the fish species sampled in the river. Disease outbreak among rural dwellers depending on the river for domestic and other uses is imminent and they lack knowledge on its health implication. Moreover, the study maintained that the micronuclei in fish could be measured from either the gill or kidney; however, gill is more efficient as it enables collection of several samples from the same individuals without sacrificing it, and Synodontis clarias fish species appeared to be more vulnerable to the genotoxic damage than Tilapia nilotica. Consequently, the study recommended regular monitoring (micronucleus tests) of edible aquatic life such as Synodontis clarias in order to eliminate the danger of people feeding on toxic metals, some of which are carcinogenic. Copyright © 2014 Elsevier B.V. All rights reserved.

  8. Mutagenic effects of tributyltin and inorganic lead (Pb II on the fish H. malabaricus as evaluated using the comet assay and the piscine micronucleus and chromosome aberration tests

    Directory of Open Access Journals (Sweden)

    Ferraro Marcos Vinícius M.

    2004-01-01

    Full Text Available Genotoxicity studies on toxic metals and their organic compounds are very important, especially so in the investigation of the effects of these compounds on the aquatic environments where they tend to accumulate. The use of endemic aquatic organisms as biological sentinels has proved useful to environmental monitoring. We assessed the mutagenic potential of tributyltin (TBT and inorganic lead (PbII using samples of the fish Hoplias malabaricus (commonly called traíra using the comet assay and the piscine micronucleus and chromosome aberration tests. Eighteen H. malabaricus were acclimatized in three individual aquariums, each containing six fish, six fish being exposed to 0.3 mg/g of body weight (bw of TBT, six to 21 mg/g bw of PbII and six being used as controls. Exposure to TBT and PbII was achieved by feeding the fish every five days with Astyanax (a small fish that is part of the normal diet of H. malabaricus which had been injected with solutions of TBT, PbII or with water (the control group. After two months the H. malabaricus were sacrificed and their peripheral blood collected and subjected to the comet and micronucleus assays, the chromosome aberration assay being conducted using kidney-tissue. Although the comet assay showed now mutagenic effects at the lead concentrations used but encountered results with TBT, the micronucleus and chromosome aberrations assays both indicated that TBT and PbII are potentially mutagenic (p < 0.01, the micronucleus assay showing morphological alterations of the nucleus.

  9. HUMN project initiative and review of validation, quality control and prospects for further development of automated micronucleus assays using image cytometry systems

    Czech Academy of Sciences Publication Activity Database

    Fenech, M.; Kirsch-Volders, M.; Rössnerová, Andrea; Šrám, Radim; Romm, H.; Bolognesi, C.; Ramakumar, A.; Soussaline, F.; Schunck, CH.; Elhajouji, A.; Anwar, W.; Bonassi, S.

    2013-01-01

    Roč. 216, č. 5 (2013), s. 541-552 ISSN 1438-4639 R&D Projects: GA ČR GAP503/11/0084 Grant - others:Project NewGenesis(XE) FOOD -CT-2005-016320 Institutional support: RVO:68378041 Keywords : Micronucleus * Cytokinesis-block * Automation Subject RIV: DN - Health Impact of the Environment Quality Impact factor: 3.276, year: 2013

  10. Evaluation of the repeated-dose liver micronucleus assay using 2,4-dinitrotoluene: a report of a collaborative study by CSGMT/JEMS.MMS.

    Science.gov (United States)

    Maeda, Akihisa; Tsuchiyama, Hiromi; Asaoka, Yoshiji; Hirakata, Mikito; Miyoshi, Tomoya; Oshida, Keiyu; Miyamoto, Yohei

    2015-03-01

    The liver micronucleus assay using young adult rats has the potential to detect liver carcinogens by repeated dosing, and could be expected to be integrated into repeated-dose toxicity studies using a hepatocyte isolation method without the traditional in situ collagenase perfusion. In this study, to assess the performance of the repeated-dose liver micronucleus assay, 2,4-dinitrotoluene (DNT), which is a rodent liver carcinogen, was administered orally to male rats at doses of 50, 100 and 200 mg/kg/day once daily for 14 or 28 consecutive days, and the frequencies of micronucleated hepatocytes (MNHEPs) and micronucleated immature erythrocytes (MNIMEs) were examined. Significant increases in the MNHEPs were observed at 50 mg/kg/day or more in the 14-day treatment, and 50 and 100 mg/kg/day in the 28-day treatment. These increases were dependent on both the dose and the number of administrations, which indicates the possibility that the MNHEPs accumulate as a result of repeated dosing. In contrast, no increase in the MNIMEs was observed. In conclusion, the repeated-dose liver micronucleus assay using young adult rats is sufficiently sensitive to detect the genotoxicity of 2,4-DNT at a low dose.

  11. The effect of gamma radiation on the Common carp (Cyprinus carpio): In vivo genotoxicity assessment with the micronucleus and comet assays.

    Science.gov (United States)

    M K, Praveen Kumar; Soorambail K, Shyama; Bhagatsingh Harisingh, Sonaye; D'costa, Avelyno; Ramesh Chandra, Chaubey

    2015-10-01

    Radioactive wastes may be leached into freshwater, either accidentally or in industrial effluents. We have studied gamma radiation-induced DNA damage in the freshwater fish Cyprinus carpio. Fish were irradiated with 2-10Gy gamma radiation and genotoxic effects in blood cells were studied with the micronucleus (MN) and comet assays. Micronuclei and a dose-dependent increase in comet-tail DNA were seen in dose- and time-dependent studies. The highest % tail DNA was observed at 24h, declining until 72h, which may indicate the repair of radiation-induced DNA single-strand breaks after gamma radiation. However, double-stranded DNA damage may not have been repaired, as indicated by increased micronuclei at later periods. A positive correlation was observed between the comet and micronucleus assay results. This study confirms the mutagenic/genotoxic potential of gamma radiation in the Common carp, as well as the possible combined use of the micronucleus and comet assays for in vivo laboratory studies with fresh-water fish for screening the genotoxic potential of radioactive pollution. Copyright © 2015 Elsevier B.V. All rights reserved.

  12. Ultrastructural changes of the UV-irradiated micronucleus in vegetative cells of paramecium bursaria and its functional importance

    International Nuclear Information System (INIS)

    Borkhsenius, O.N.; Fokin, S.I.

    1982-01-01

    Ultrastructural micronucleus (MI) changes of vegetative cells in Paramecium bursaria in 0.5-7 h after MI ultraviolet irradiation and cell posterity with irradiated MI after different periods (2.6 and 30 days, three years) after ultraviolet irradiation have been studied. It is established that MI irradiation at a dose of 306J/m 2 doesn't result in its loss in postradiation generations however in posterity MI considerable ultrastructural changes occur. In two days after operation the shell of descendant MI of irradiated Paramecium bursaria forms multiple blades; small chromatin blocks considerably increase in sizes and, as a rule, occupy the central position in a nucleus. During later periods afeter irradiation (6 days) density of chromatin elements in MI begins to change. By the 30-st day in MI fine-fibrillar karyoplasm detected are only not numerous chromatin structures scattered in disorder. The results obtained point to the existence of the ''cryptic'' type MIs are not revealed at the light-optical level but preserved in a series of postradiation generations. The presence of such MIs in viable cultures of P. bursaria confirms indirectly MI significance in vegetatic life of P. bursaria

  13. The Tradescantia micronucleus assay is a highly sensitive tool for the detection of low levels of radioactivity in environmental samples.

    Science.gov (United States)

    Mišík, Miroslav; Krupitza, Georg; Mišíková, Katarina; Mičieta, Karol; Nersesyan, Armen; Kundi, Michael; Knasmueller, Siegfried

    2016-12-01

    Environmental contamination with radioactive materials of geogenic and anthropogenic origin is a global problem. A variety of mutagenicity test procedures has been developed which enable the detection of DNA damage caused by ionizing radiation which plays a key role in the adverse effects caused by radioisotopes. In the present study, we investigated the usefulness of the Tradescantia micronucleus test (the most widely used plant based genotoxicity bioassay) for the detection of genetic damage caused by environmental samples and a human artifact (ceramic plate) which contained radioactive elements. We compared the results obtained with different exposure protocols and found that direct exposure of the inflorescences is more sensitive and that the number of micronuclei can be further increased under "wet" conditions. The lowest dose rate which caused a significant effect was 1.2 μGy/h (10 h). Comparisons with the results obtained with other systems (i.e. with mitotic cells of higher plants, molluscs, insects, fish and human lymphocytes) show that the Tradescantia MN assay is one to three orders of magnitude more sensitive as other models, which are currently available. Taken together, our findings indicate that this method is due to its high sensitivity a unique tool, which can be used for environmental biomonitoring in radiation polluted areas. Copyright © 2016 Elsevier Ltd. All rights reserved.

  14. Ultrastructural changes of the UV-irradiated micronucleus in vegetative cells of Paramecium bursaria and its functional importance

    Energy Technology Data Exchange (ETDEWEB)

    Borkhsenius, O.N.; Fokin, S.I. (Leningradskij Gosudarstvennyj Univ. (USSR). Biologicheskij Nauchno-Issledovatel' skij Inst.)

    1982-01-01

    Ultrastructural micronucleus (MI) changes of vegetative cells in Paramecium bursaria in 0.5-7 h after MI ultraviolet irradiation and cell posterity with irradiated MI after different periods (2.6 and 30 days, three years) after ultraviolet irradiation have been studied. It is established that MI irradiation at a dose of 306J/m/sup 2/ doesn't result in its loss in postradiation generations however in posterity MI considerable ultrastructural changes occur. In two days after operation the shell of descendant MI of irradiated Paramecium bursaria forms multiple blades; small chromatin blocks considerably increase in sizes and, as a rule, occupy the central position in a nucleus. During later periods after irradiation (6 days) density of chromatin elements in MI begins to change. By the 30th day in MI fine-fibrillar karyoplasm detected are only not numerous chromatin structures scattered in disorder. The results obtained point to the existence of the ''cryptic'' type MIs are not revealed at the light-optical level but preserved in a series of postradiation generations. The presence of such MIs in viable cultures of P. bursaria confirms indirectly MI significance in vegetatic life of P. bursaria.

  15. Micronucleus frequency and hematologic index in Colossoma macropomum (Pisces, Ariidae) for environmental impact assessment at a protected area in Brazil

    International Nuclear Information System (INIS)

    Sousa, Debora Batista Pinheiro; Neta, Raimunda Nonata Fortes Carvalho

    2014-01-01

    This study used micronucleus assays and erythrocyte indices in the freshwater fish tambaqui, Colossoma macropomum, to assess environmental impacts in the Environmental Protection Area at Maracanã, São Luis, Brazil. Fish were sampled from two locations within the protected area, Serena Lagoon and Ambude River, on four occasions. Biometric data (length and weight) and an aliquot of blood were collected from each fish for analysis. Erythrocyte indices including: mean corpuscular volume, mean corpuscular hemoglobin and mean corpuscular hemoglobin concentration were calculated, and blood samples were examined for micronuclei and nuclear morphological changes. Micronuclei were found in fish from both locations, although the frequency was higher in fish from Ambude River. Nuclear morphological changes were identified only in fish collected from Ambude River. Several nuclear morphological changes were found in erythrocytes stained with Giemsa, including: micronuclei and binucleate nuclei. On average, erythrocyte indices were lower in fish collected from Ambude River than in those from Serena Lagoon. Our results indicate that micronuclei and erythrocyte indices can be used in C. macropomum as indicators of environmental health

  16. Evaluating the genotoxic effects of workers exposed to lead using micronucleus assay, comet assay and TCR gene mutation test

    International Nuclear Information System (INIS)

    Chen Zhijian; Lou Jianlin; Chen Shijie; Zheng Wei; Wu Wei; Jin Lifen; Deng Hongping; He Jiliang

    2006-01-01

    To evaluate the genotoxic effects of lead (Pb) exposure, 25 workers in a workplace producing storage battery were monitored for three genetic end-points using micronucleus (MN) assay, comet assay and TCR gene mutation test. Twenty-five controls were matched with workers according to age, gender and smoking. The air Pb concentration in the workplace was 1.26 mg/m 3 . All subjects were measured for Pb concentration of blood by atom absorption spectrophotometry. The mean Pb concentration of blood in workers (0.32 mg/l) was significantly higher than that in controls (0.02 mg/l). The results of MN test showed that the mean micronuclei rate (MNR) and mean micronucleated cells rate (MCR) in workers were 9.04 ± 1.51 per mille and 7.76 ± 1.23 per mille , respectively, which were significantly higher than those (2.36 ± 0.42 per mille and 1.92 ± 0.31 per mille ) in controls (P -4 and 1.74 ± 0.17 x 10 -4 , respectively, there was no significant difference between workers and controls (P > 0.05). The results of our study indicated that the genetic damage was detectable in 25 workers occupationally exposed to lead

  17. Micronucleus frequency and hematologic index in Colossoma macropomum (Pisces, Ariidae) for environmental impact assessment at a protected area in Brazil

    Energy Technology Data Exchange (ETDEWEB)

    Sousa, Debora Batista Pinheiro, E-mail: deborabpsousa@gmail.com [Postgraduate Program of Aquatic Resources and Fishery (PPGRAP/UEMA), State University of Maranhão (Brazil); Neta, Raimunda Nonata Fortes Carvalho [Department of Chemistry and Biology, State University of Maranhão (Brazil)

    2014-10-06

    This study used micronucleus assays and erythrocyte indices in the freshwater fish tambaqui, Colossoma macropomum, to assess environmental impacts in the Environmental Protection Area at Maracanã, São Luis, Brazil. Fish were sampled from two locations within the protected area, Serena Lagoon and Ambude River, on four occasions. Biometric data (length and weight) and an aliquot of blood were collected from each fish for analysis. Erythrocyte indices including: mean corpuscular volume, mean corpuscular hemoglobin and mean corpuscular hemoglobin concentration were calculated, and blood samples were examined for micronuclei and nuclear morphological changes. Micronuclei were found in fish from both locations, although the frequency was higher in fish from Ambude River. Nuclear morphological changes were identified only in fish collected from Ambude River. Several nuclear morphological changes were found in erythrocytes stained with Giemsa, including: micronuclei and binucleate nuclei. On average, erythrocyte indices were lower in fish collected from Ambude River than in those from Serena Lagoon. Our results indicate that micronuclei and erythrocyte indices can be used in C. macropomum as indicators of environmental health.

  18. Techniques for induction of premature chromosome condensation (PCC) by Calyculin-A and micronucleus assay for biodosimetry in Vietnam

    International Nuclear Information System (INIS)

    Pham Ngoc Duy; Tran Que; Hoang Hung Tien; Bui Thi Kim Luyen; Nguyen Thi Kim Anh; Ha Thi Ngoc Lien

    2014-01-01

    The International Atomic Energy Agency (IAEA) and World Health Organization are interested in biological dosimetry method for radiation emergency medicine currently. Some cytogenetic techniques such as premature chromosome condensation (PCC) induced by Calyculin-A and micronucleus (MN) assay are necessary to develop biodosimetry in Vietnam. In this study, we optimized the condition for MN assay with 6 µg/ml Cytochalasin-B concentration and 72.5 hours for peripheral lymphocyte blood culture. The optimization for PCC method is 50 nM Calyculin-A concentration for 45 minutes peripheral lymphocyte blood treatment. For samples exposed to 3.0 Gy gamma 60 Co (dose rate 0.0916 Gy/s), the frequency of MN is 19.02 ± 0.38%, NBP is 1.95 ± 0.28%, dicentric and ring is 41.43 ± 8.12% and frag and min is 63.33 ± 5.16%. For samples exposed to 6.0 Gy gamma 60 Co (dose rate 0.0916 Gy/s), the frequency of ring-PCC is 17.73 ± 2.46%, extra unite is 218.91± 7.58%, dicentric is 83.81 ± 1.09%, ring is 10.75 ± 1.74%, fragment and minute is 193.17 ± 13.10%. MN and ring-PCC are specific marker applying for biodosimetry. (author)

  19. Cytogenetic radiosensitivity of G0-lymphocytes of breast and esophageal cancer patients as determined by micronucleus assay

    International Nuclear Information System (INIS)

    Mozdarani, H.; Mansouri, Z.; Haeri, S.A.

    2005-01-01

    Enhanced chromosomal radiosensitivity is a feature of many cancer predisposition conditions, indicative of the important role of chromosomal alterations in carcinogenesis. In this study the cytokinesis-blocked micronucleous assay was used to compare the radiosensitivity of blood lymphocytes obtained from Iranian breast or esophageal cancer patients (n=50, n=16; respectively) with that of control individuals (n=40). For each sample, one thousand binucleate lymphocytes were analyzed before and after in vitro exposure to 3 Gy of γ rays. The radiation-induced frequency of micronucleus was significantly higher in the breast cancer group (261/1,000 binucleated cells) than in esophageal cancer group (241/1,000 binucleated cells, P<0.01) or in the control group (240/1,000 binucleated cells, P<0.01). The results indicate that breast cancer patients are more radiosensitive compared to normal healthy individuals or esophageal cancer patients. Increased radiosensitivity could be due to defects in DNA repair genes involved in breast cancer formation. Since patients with esophageal cancer did not show elevated radiosensitivity, it is assumed that the contribution of radiosensitivity-related genes to the development of esophageal cancer may be smaller than the contribution of those genes to breast cancer. (author)

  20. 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

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

    Directory of Open Access Journals (Sweden)

    Antje eFröhling

    2015-09-01

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

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

    Science.gov (United States)

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

    2016-01-01

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

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

    International Nuclear Information System (INIS)

    Marcusson-Staahl, Maritha; Cederbrant, Karin

    2003-01-01

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

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

    Science.gov (United States)

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

    2014-07-01

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

  5. How to score questionnaires

    NARCIS (Netherlands)

    Hofstee, W.K.B.; Ten Berge, J.M.F.; Hendriks, A.A.J.

    The standard practice in scoring questionnaires consists of adding item scores and standardizing these sums. We present a set of alternative procedures, consisting of (a) correcting for the acquiescence variance that disturbs the structure of the questionnaire; (b) establishing item weights through

  6. SCORE - A DESCRIPTION.

    Science.gov (United States)

    SLACK, CHARLES W.

    REINFORCEMENT AND ROLE-REVERSAL TECHNIQUES ARE USED IN THE SCORE PROJECT, A LOW-COST PROGRAM OF DELINQUENCY PREVENTION FOR HARD-CORE TEENAGE STREET CORNER BOYS. COMMITTED TO THE BELIEF THAT THE BOYS HAVE THE POTENTIAL FOR ETHICAL BEHAVIOR, THE SCORE WORKER FOLLOWS B.F. SKINNER'S THEORY OF OPERANT CONDITIONING AND REINFORCES THE DELINQUENT'S GOOD…

  7. The Bandim tuberculosis score

    DEFF Research Database (Denmark)

    Rudolf, Frauke; Joaquim, Luis Carlos; Vieira, Cesaltina

    2013-01-01

    Background: This study was carried out in Guinea-Bissau ’ s capital Bissau among inpatients and outpatients attending for tuberculosis (TB) treatment within the study area of the Bandim Health Project, a Health and Demographic Surveillance Site. Our aim was to assess the variability between 2...... physicians in performing the Bandim tuberculosis score (TBscore), a clinical severity score for pulmonary TB (PTB), and to compare it to the Karnofsky performance score (KPS). Method : From December 2008 to July 2009 we assessed the TBscore and the KPS of 100 PTB patients at inclusion in the TB cohort and...

  8. Volleyball Scoring Systems.

    Science.gov (United States)

    Calhoun, William; Dargahi-Noubary, G. R.; Shi, Yixun

    2002-01-01

    The widespread interest in sports in our culture provides an excellent opportunity to catch students' attention in mathematics and statistics classes. One mathematically interesting aspect of volleyball, which can be used to motivate students, is the scoring system. (MM)

  9. Age and smoking habit influence on the spontaneous and radiation induced frequencies of the micronucleus in human lymphocytes

    International Nuclear Information System (INIS)

    Di Giorgio, M.; Nasazzi, N.; Heredia, M.L.

    1996-01-01

    Several endpoints have been used for monitoring human population that have been exposed at work or in the environment to genotoxic agents, particularly to ionizing radiation. The cytokinesis-block micronucleus (MN) assay in peripheral lymphocytes is a reliable method for evaluating radiation induced chromosomal damage (DNA breaks and mitotic spindle disturbances) and thus, a suitable dosimeter for estimating in vivo whole body exposures. A research to determine the influence of age, sex and life style factors (smoking habits) on the MN spontaneous and radiation induced frequencies was carried out in order to define the use of this assay in Biological Dosimetry. The estimation of MN frequencies was analyzed in lymphocytes cultures from 50 health donors aged between 4 and 60 years. Based on the smoking habits, they were divided into 2 groups. A fraction of the sample was irradiated in vitro with γ-rays in the range of 0.35 Gy to 4 Gy. A statistically significant influence on the spontaneous MN frequency was observed (R 2 = 0.59) when the variables age and smoking habit were analyzed, and a statistically significant influence on the radiation induced MN frequency was also obtained (R 2 = 0.86) when dose, age and smoking habit were studied. Sex did not influence significantly MN variability, but there was a greater dispersion in the results obtained from female donors, when compared to males, possibly due to the loss of X chromosomes. The comparison of the data from smoking donors to the data from non smoking donors supports the convenience of taking into account the smoking habit for estimating in vivo whole body exposure to γ-rays for doses below 2 Gy. (authors). 8 refs., 3 figs., 2 tabs

  10. Evidence of heritable lethal mutations in progeny of X-irradiated CHO cells by micronucleus count in clon-cells

    International Nuclear Information System (INIS)

    Hagemann, G.; Kreczik, A.; Treichel, M.

    1996-01-01

    Low doses of ionizing radiation reduce the growth rates of clones following irradiation of the progenitor cells. Such reductions of clone growth have been proven by means of measurements of clone size distributions. The medians of such distributions can be used to quantify the radiation damage. Prolongations of generation times and cell death as result of heritable lethal mutations have been discussed as causes for the reduction of clone growth. The cell number of a clone of hypotetraploid CHO-cells was compared to the frequency of micronucleated binucleated cells in the same clone using the cytokinesis-block-micronucleus method. The dose dependent reduction of clone sizes is measured by the difference of the medians (after log transformation) of the clone size distributions. At cytochalasin-B concentrations of 1 μg/ml and after an incubation time of 16 h a yield of binucleated cells of about 50% was obtained. Median clone size differences as a measure of clonal radiation damage increased linearly with incubation times of 76, 100, 124, and 240 h following irradiation with 3, 5, 7, and 12 Gy. The frequency of binucleated clone cells with micronuclei strongly increased with decreasing clone size by a factor up to 20 following irradiation with 3, 5, and 7 Gy. The frequency of micronucleated binucleated clone cells was found to be independent of incubation time after irradiation. Radiation induced clone size reductions result from cell losses caused by intraclonal expression of micronuclei which have its origin in heritable lethal mutations. Measurements of clone size distributions can be done automatically. They can serve as predictive test for determination of median cell loss rates of surviving cell clones. (orig./MG) [de

  11. Dose-Response Assessment of Four Genotoxic Chemicals in a Combined Mouse and Rat Micronucleus and Comet Assay Protocol

    Science.gov (United States)

    Recio, Leslie; Hobbs, Cheryl; Caspary, William; Witt, Kristine L.

    2012-01-01

    The in vivo micronucleus (MN) assay has proven to be an effective measure of genotoxicity potential. However, sampling a single tissue (bone marrow) for a single indicator of genetic damage using the MN assay provides a limited genotoxicity profile. The in vivo alkaline (pH>13) Comet assay, which detects a broad spectrum of DNA damage, can be applied to a variety of rodent tissues following administration of test agents. To determine if the Comet assay is a useful supplement to the in vivo MN assay, a combined test protocol (MN/Comet assay) was conducted in male B6C3F1 mice and F344/N rats using four model genotoxicants: ethyl methanesulfonate (EMS), acrylamide (ACM), cyclophosphamide (CP), and vincristine sulfate (VS). Test compounds were administered on 4 consecutive days at 24-hour intervals (VS was administered to rats for 3 days); animals were euthanized 4 hours after the last administration. All compounds induced significant increases in micronucleated reticulocytes (MN-RET) in the peripheral blood of mice, and all but ACM induced MN-RET in rats. EMS and ACM induced significant increases in DNA damage, measured by the Comet assay, in multiple tissues of mice and rats. CP-induced DNA damage was detected in leukocytes and duodenum cells. VS, a spindle fiber disrupting agent, was negative in the Comet assay. Based on these results, the MN/Comet assay holds promise for providing more comprehensive assessments of potential genotoxicants, and the National Toxicology Program is presently using this combined protocol in its overall evaluation of the genotoxicity of substances of public health concern. PMID:20371966

  12. Evaluation of spontaneous and radiation-induced micronucleus frequency in cultured human peripheral blood lymphocytes depending on age and sex

    Energy Technology Data Exchange (ETDEWEB)

    Jeon, H. J.; Kang, C. M.; Chung, H. C. [Korea Institute of Radiological and Medical Sciences, Seoul (Korea, Republic of)] (and others)

    2002-12-15

    The goal of this study was to provide data on the dose-dependent production of MicroNucleus (MN) in human lymphocytes irradiated with {sup 60}Co {gamma}-rays and 50MeV neutron, and to evaluate predictive markers of intrinsic radiosensitivity in individuals for monitoring occupational or environmental radiation exposure. For the dose-response study, heparinized whole blood of 10 healthy volunteers was irradiated with {sup 60}Co {gamma}-rays employing of 0.25-8Gy. The MNs were observed all doses, and the numerical changes according to doses. In dose-response curves fit linear-quadratic form (alpha =0.31{+-}0.049, beta =0.0022{+-}0.0022) for {gamma}-rays, but (alpha=0.99{+-}0.528, beta =0.0093{+-}0.0047) for neutron. Neutrons were than {gamma}-rays effective in producing MN with dose-dependent manner. The frequency of MN varies with dose. The RBE (Relative Biological Effectiveness) for micronuclei was 2.370.17. Further studies were carried out to provide evidence for the existence of individual variations in age-dependent responses to radiation. Spontaneous and radiation-induced MN varies greatly among individuals, and little is known about the molecular mechanisms of this variability. It was shown that the increased level of spontaneous cell with MN was observed with increasing age. The relationship between radiosensitivity and the increased spontaneous level of MN may be in inverse proportion. These studies indicated that the MN assay have a high potential as a rapid, sensitive and accurate method which can be used to monitor a large population exposed to radiation for rapid triage in the case of a large-scale accident.

  13. Evaluation of spontaneous and radiation-induced micronucleus frequency in cultrued human peripheral blood lymphocytes depending on age and sex

    Energy Technology Data Exchange (ETDEWEB)

    Jeon, H. J.; Kang, C. M.; Chung, H. C. [Korea Cancer Center Hospital, Seoul (Korea, Republic of)] [and others

    2002-07-01

    The goal of this study was to provide data on the dose-dependent production of micronucleus (MN) in human lymphocytes irradiated with {sub 60} Co {gamma} -rays and 50MeV neutron, and to evaluate predictive markers of intrinsic radiosensitivity in individuals for monitoring occupational or environmental radiation exposure. For the dose-response study, heparinized whole blood of 10 healthy volunteers was irradiated with {sub 60} Co {gamma} -rays employing of 0.25-8Gy. The MNs were observed all doses, and the numerical changes according to doses. In dose-response curves fit linear- quadratic form (alpha =0.31{+-}0.049, beta =0.0022{+-}0.0022) for {gamma} -rays, but (alpha =0.99{+-}0.528, beta =0.0093{+-}0.0047) for neutron. Neutrons were than {gamma} -rays effective in producing MN with dose-dependent manner. The frequency of MN varies with dose. The RBE for micronuclei was 2.37{+-}0.17. Further studied are carried out to provide evidence for the existence of individual variations in age-dependent responses to radiation. Spontaneous and radiation-induced MN varies greatly between individuals, and little is known about the molecular mechanisms of this variability. It was shown that the increased level of spontaneous cell with MN was observed with increasing age. The relationship between radiosensitivity and the increased spontaneous level of MN may be in inverse proportion. These studies indicates that the MN assay have a high potential as a rapid, sensitive and accurate method which can be used to monitor a large population exposed to radiation for rapid triage in the case of a large-scale accident.

  14. Genotoxicity of Heterocyclic PAHs in the Micronucleus Assay with the Fish Liver Cell Line RTL-W1

    Science.gov (United States)

    Brinkmann, Markus; Blenkle, Henning; Salowsky, Helena; Bluhm, Kerstin; Schiwy, Sabrina; Tiehm, Andreas; Hollert, Henner

    2014-01-01

    Heterocyclic aromatic hydrocarbons are, together with their un-substituted analogues, widely distributed throughout all environmental compartments. While fate and effects of homocyclic PAHs are well-understood, there are still data gaps concerning the ecotoxicology of heterocyclic PAHs: Only few publications are available investigating these substances using in vitro bioassays. Here, we present a study focusing on the identification and quantification of clastogenic and aneugenic effects in the micronucleus assay with the fish liver cell line RTL-W1 that was originally derived from rainbow trout (Oncorhynchus mykiss). Real concentrations of the test items after incubation without cells were determined to assess chemical losses due to, e.g., sorption or volatilization, by means of gas chromatography-mass spectrometry. We were able to show genotoxic effects for six compounds that have not been reported in vertebrate systems before. Out of the tested substances, 2,3-dimethylbenzofuran, benzothiophene, quinoline and 6-methylquinoline did not cause substantial induction of micronuclei in the cell line. Acridine caused the highest absolute induction. Carbazole, acridine and dibenzothiophene were the most potent substances compared with 4-nitroquinoline oxide, a well characterized genotoxicant with high potency used as standard. Dibenzofuran was positive in our investigation and tested negative before in a mammalian system. Chemical losses during incubation ranged from 29.3% (acridine) to 91.7% (benzofuran) and may be a confounding factor in studies without chemical analyses, leading to an underestimation of the real potency. The relative potency of the investigated substances was high compared with their un-substituted PAH analogues, only the latter being typically monitored as priority or indicator pollutants. Hetero-PAHs are widely distributed in the environment and even more mobile, e.g. in ground water, than homocyclic PAHs due to the higher water solubility. We

  15. Correlation of In Vivo Versus In Vitro Benchmark Doses (BMDs) Derived From Micronucleus Test Data: A Proof of Concept Study.

    Science.gov (United States)

    Soeteman-Hernández, Lya G; Fellows, Mick D; Johnson, George E; Slob, Wout

    2015-12-01

    In this study, we explored the applicability of using in vitro micronucleus (MN) data from human lymphoblastoid TK6 cells to derive in vivo genotoxicity potency information. Nineteen chemicals covering a broad spectrum of genotoxic modes of action were tested in an in vitro MN test using TK6 cells using the same study protocol. Several of these chemicals were considered to need metabolic activation, and these were administered in the presence of S9. The Benchmark dose (BMD) approach was applied using the dose-response modeling program PROAST to estimate the genotoxic potency from the in vitro data. The resulting in vitro BMDs were compared with previously derived BMDs from in vivo MN and carcinogenicity studies. A proportional correlation was observed between the BMDs from the in vitro MN and the BMDs from the in vivo MN assays. Further, a clear correlation was found between the BMDs from in vitro MN and the associated BMDs for malignant tumors. Although these results are based on only 19 compounds, they show that genotoxicity potencies estimated from in vitro tests may result in useful information regarding in vivo genotoxic potency, as well as expected cancer potency. Extension of the number of compounds and further investigation of metabolic activation (S9) and of other toxicokinetic factors would be needed to validate our initial conclusions. However, this initial work suggests that this approach could be used for in vitro to in vivo extrapolations which would support the reduction of animals used in research (3Rs: replacement, reduction, and refinement). © The Author 2015. Published by Oxford University Press on behalf of the Society of Toxicology.

  16. In situ monitoring of urban air in Córdoba, Argentina using the Tradescantia-micronucleus (Trad-MCN) bioassay

    Science.gov (United States)

    Carreras, H. A.; Pignata, M. L.; Saldiva, P. H. N.

    During the last decades, a significant deterioration of ambient air quality has been observed in Argentina. However, the availability of air pollution monitoring stations is still limited to only few cities. In this study, we investigated the genotoxicity of ambient levels of air pollution in Córdoba using the Tradescantia micronucleus assay. The experiment was performed from October, 2004 to April 2005. Pots with Tradescantia pallida were placed in three sites: Córdoba city center, characterized by important avenues with high traffic activity (cars, taxis, and public transport vehicles); the university campus, along a side road with heavy traffic of gasoline and diesel powered vehicles, buses and trucks; and a residential area, with no significant local sources of air pollution. Twenty young T. pallida inflorescences were collected from each sampling site in November, February and April. Micronuclei frequencies were determined in early tetrads of pollen mother cells and expressed as MCN/100 tetrads. Simultaneously, the environmental levels of total suspended particles (24 h mean) were determined for each site. A significant difference in micronuclei frequency was observed among sites ( p=0.036). Post-hoc analysis revealed that the residential area exhibited a lower micronuclei frequency than the university and city center areas. In conclusion, we found that the gradients of ambient air pollution of Córdoba are associated with changes in the spontaneous micronuclei frequency of Tradescantia pollen mother cells. These results indicate that in situ biomonitoring with higher plants may be useful for characterizing air pollution in areas without instrumental monitoring techniques, or for exploring the distribution of air contaminants at a microscale.

  17. Genotoxicity of heterocyclic PAHs in the micronucleus assay with the fish liver cell line RTL-W1.

    Directory of Open Access Journals (Sweden)

    Markus Brinkmann

    Full Text Available Heterocyclic aromatic hydrocarbons are, together with their un-substituted analogues, widely distributed throughout all environmental compartments. While fate and effects of homocyclic PAHs are well-understood, there are still data gaps concerning the ecotoxicology of heterocyclic PAHs: Only few publications are available investigating these substances using in vitro bioassays. Here, we present a study focusing on the identification and quantification of clastogenic and aneugenic effects in the micronucleus assay with the fish liver cell line RTL-W1 that was originally derived from rainbow trout (Oncorhynchus mykiss. Real concentrations of the test items after incubation without cells were determined to assess chemical losses due to, e.g., sorption or volatilization, by means of gas chromatography-mass spectrometry. We were able to show genotoxic effects for six compounds that have not been reported in vertebrate systems before. Out of the tested substances, 2,3-dimethylbenzofuran, benzothiophene, quinoline and 6-methylquinoline did not cause substantial induction of micronuclei in the cell line. Acridine caused the highest absolute induction. Carbazole, acridine and dibenzothiophene were the most potent substances compared with 4-nitroquinoline oxide, a well characterized genotoxicant with high potency used as standard. Dibenzofuran was positive in our investigation and tested negative before in a mammalian system. Chemical losses during incubation ranged from 29.3% (acridine to 91.7% (benzofuran and may be a confounding factor in studies without chemical analyses, leading to an underestimation of the real potency. The relative potency of the investigated substances was high compared with their un-substituted PAH analogues, only the latter being typically monitored as priority or indicator pollutants. Hetero-PAHs are widely distributed in the environment and even more mobile, e.g. in ground water, than homocyclic PAHs due to the higher water

  18. Increased micronucleus, nucleoplasmic bridge, and nuclear bud frequencies in the peripheral blood lymphocytes of diesel engine exhaust-exposed workers.

    Science.gov (United States)

    Zhang, Xiao; Duan, Huawei; Gao, Feng; Li, Yuanyuan; Huang, Chuanfeng; Niu, Yong; Gao, Weimin; Yu, Shanfa; Zheng, Yuxin

    2015-02-01

    The International Agency for Research on Cancer has recently reclassified diesel engine exhaust (DEE) as a Group 1 carcinogen. Micronucleus (MN), nucleoplasmic bridge (NPB), and nuclear bud (NBUD) frequencies in peripheral blood lymphocytes (PBLs) are associated with cancer risk. However, the impact of DEE exposure on MN frequency has not been thoroughly elucidated due to mixed exposure and its impact on NPB and NBUD frequencies has never been explored in humans. We recruited 117 diesel engine testing workers with exclusive exposure to DEE and 112 non-DEE-exposed workers, and then we measured urinary levels of 4 mono-hydroxylated polycyclic aromatic hydrocarbons (OH-PAHs) using high-performance liquid chromatography-mass spectrometry as well as MN, NPB, and NBUD frequencies in PBLs using cytokinesis-block MN assay. The DEE-exposed workers exhibited significantly higher MN, NPB, and NBUD frequencies than the non-DEE-exposed workers (P < 0.05). Among all study subjects, increasing levels of all 4 urinary OH-PAHs, on both quartile and continuous scales, were associated with increased MN, NPB, and NBUD frequencies (all P < 0.05). When the associations were analyzed separately in DEE-exposed and non-DEE-exposed workers, we found that the association between increasing quartiles of urinary 9-hydroxyphenanthrene (9-OHPh) and MN frequencies persisted in DEE-exposed workers (P = 0.001). The percent of MN frequencies increased, on average, by 23.99% (95% confidential interval, 9.64-39.93) per 1-unit increase in ln-transformed 9-OHPh. Our results clearly show that exposure to DEE can induce increases in MN, NPB, and NBUD frequencies in PBLs and suggest that DEE exposure level is associated with MN frequencies. © The Author 2014. Published by Oxford University Press on behalf of the Society of Toxicology. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  19. 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.

  20. Evaluation of a liver micronucleus assay in young rats (IV): a study using a double-dosing/single-sampling method by the Collaborative Study Group for the Micronucleus Test (CSGMT)/Japanese Environmental Mutagen Society (JEMS)-Mammalian Mutagenicity Study Group (MMS).

    Science.gov (United States)

    Takasawa, Hironao; Suzuki, Hiroshi; Ogawa, Izumi; Shimada, Yasushi; Kobayashi, Kazuo; Terashima, Yukari; Matsumoto, Hirotaka; Oshida, Keiyu; Ohta, Ryo; Imamura, Tadashi; Miyazaki, Atsushi; Kawabata, Masayoshi; Minowa, Shigenori; Maeda, Akihisa; Hayashi, Makoto

    2010-04-30

    A collaborative study was conducted to evaluate whether a liver micronucleus assay using four-week-old male F344 rats can be used to detect genotoxic rat hepatocarcinogens using double-dosing with a single-sampling 4 days after the second dose. The assay methods were thoroughly validated by the seven laboratories involved in the study. Seven chemicals, 2,4-diaminotoluene, diethyl nitrosamine, p-dimethylaminoazobenzene, 1,2-dimethylhydrazine dihydrochloride, 2,4-dinitrotolunene, 2,6-dinitrotoluene and mitomycin C, known to produce positive responses in the single-dosing/triple-sampling method were selected for use in the present study, and each chemical was examined in two laboratories with the exception of 2,4-dinitrotolunene. Although several of the compounds were examined at lower doses for reasons of toxicity than in the single-dosing/triple-sampling method, all chemicals tested in the present study induced micronuclei in liver cells indicating a positive result. These findings suggest that the liver micronucleus assay can be used in young rats to detect genotoxic rat hepatocarcinogens using a double-dosing/single-sampling procedure. Further, the number of animals used in the liver micronucleus assay can be reduced by one-third to a half by using the double-dosing/single-sampling method. This reduction in animal numbers also has significant savings in time and resource for liver perfusion and hepatocyte isolation. Copyright 2010 Elsevier B.V. All rights reserved.

  1. 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.

  2. 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.

    Science.gov (United States)

    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.

  3. Instant MuseScore

    CERN Document Server

    Shinn, Maxwell

    2013-01-01

    Get to grips with a new technology, understand what it is and what it can do for you, and then get to work with the most important features and tasks. Instant MuseScore is written in an easy-to follow format, packed with illustrations that will help you get started with this music composition software.This book is for musicians who would like to learn how to notate music digitally with MuseScore. Readers should already have some knowledge about musical terminology; however, no prior experience with music notation software is necessary.

  4. 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.

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

    Science.gov (United States)

    Suzuki, Hiroshi; Toyooka, Tatsushi; Ibuki, Yuko

    2007-04-15

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

  6. 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

  7. 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.)

  8. 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.

  9. 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.

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

    Directory of Open Access Journals (Sweden)

    Zofia Sulowska

    2005-01-01

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

  11. 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

  12. 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.

  13. The lod score method.

    Science.gov (United States)

    Rice, J P; Saccone, N L; Corbett, J

    2001-01-01

    The lod score method originated in a seminal article by Newton Morton in 1955. The method is broadly concerned with issues of power and the posterior probability of linkage, ensuring that a reported linkage has a high probability of being a true linkage. In addition, the method is sequential, so that pedigrees or lod curves may be combined from published reports to pool data for analysis. This approach has been remarkably successful for 50 years in identifying disease genes for Mendelian disorders. After discussing these issues, we consider the situation for complex disorders, where the maximum lod score (MLS) statistic shares some of the advantages of the traditional lod score approach but is limited by unknown power and the lack of sharing of the primary data needed to optimally combine analytic results. We may still learn from the lod score method as we explore new methods in molecular biology and genetic analysis to utilize the complete human DNA sequence and the cataloging of all human genes.

  14. The Bayesian Score Statistic

    NARCIS (Netherlands)

    Kleibergen, F.R.; Kleijn, R.; Paap, R.

    2000-01-01

    We propose a novel Bayesian test under a (noninformative) Jeffreys'priorspecification. We check whether the fixed scalar value of the so-calledBayesian Score Statistic (BSS) under the null hypothesis is aplausiblerealization from its known and standardized distribution under thealternative. Unlike

  15. South African Scoring System

    African Journals Online (AJOL)

    2014-11-18

    Nov 18, 2014 ... for 80% (SASS score) and 75% (NOT) of the variation in the regression model. Consequently, SASS ... further investigation: spatial analyses of macroinvertebrate assemblages; and the use of structural and functional metrics. Keywords: .... conductivity levels was assessed using multiple linear regres- sion.

  16. Developing Scoring Algorithms

    Science.gov (United States)

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

  17. Effect of smoking habit on the frequency of micronuclei in human lymphocytes: results from the human micronucleus project

    International Nuclear Information System (INIS)

    Bonassi, Stefano; Neri, M.; Lando, Cecilia

    2004-01-01

    Full text: The effect of tobacco smoking on the frequency of micronuclei (MN) in human lymphocytes has been the object of many population studies. In most reports, the results were unexpectedly negative, and in many instances smokers had lower frequencies of MN than non-smokers. A pooled re-analysis of 24 databases from the HUMN international collaborative project has been performed with the aim of understanding the impact of smoking habits on MN frequency. The complete database included 5710 subjects, with 3501 non-smokers, 1409 current smokers, and 800 former smokers, among subjects in occupational and environmental surveys. The overall result of the re-analysis confirmed the small decrease of MN frequencies in current smokers (frequency ratio (FR =0.97, 95% confidence interval (CI =0.93-1.01) and in former smokers (FR =0.96, 95% CI =0.91-1.01), when compared to non-smokers. MN frequency was not influenced by the number of cigarettes smoked per day among subjects occupationally exposed to genotoxic agents, whereas a typical U-shaped curve is observed for non-exposed smokers, showing a significant increase of MN frequency in individuals smoking 30 cigarettes or more per day (FR =1.59, 95% CI =1.35-1.88). This analysis confirmed that smokers do not experience an overall increase in MN frequency, although when the interaction with occupational exposure is taken into account, heavy smokers were the only group showing a significant increase in genotoxic damage as measured by the micronucleus assay in lymphocytes. From these results some general recommendations for the design of bio monitoring studies involving smokers can be formulated. Quantitative data about smoking habit should always be collected because, in the absence of such data, the simple comparison of smokers versus non-smokers could be misleading. The sub-group of heavy smokers (≥30 cigarettes per day) should be specifically evaluated whenever it is large enough to satisfy statistical requirements. The

  18. Application of the micronucleus test and comet assay in Trachemys callirostris erythrocytes as a model for in situ genotoxic monitoring.

    Science.gov (United States)

    Zapata, Lina M; Bock, Brian C; Orozco, Luz Yaneth; Palacio, Jaime A

    2016-05-01

    Trachemys callirostris is a turtle species endemic to northern South America. In northern Colombia it occurs in the middle and lower Magdalena River drainage and its principal tributaries (lower Cauca and San Jorge rivers) and in other minor drainages such as the lower Sinú River. In recent years, industrial, agricultural, and mining activities have altered natural habitats in Colombia where this species occurs, and many of the pollutants released there are known to induce genetic alterations in wildlife species. The micronucleus test and comet assay are two of the most widely used methods to characterize DNA damage induced by physical and chemical agents in wildlife species, but have not been employed previously for genotoxic evaluations in T. callirostris. The goal of this study was to optimize these genotoxic biomarkers for T. callirostris erythrocytes in order to establish levels of DNA damage in this species and thereby evaluate its potential as a sentinel species for monitoring genotoxic effects in freshwater environments in northern Colombia. Both genotoxic techniques were applied on peripheral blood erythrocytes from 20 captive-reared T. callirostris individuals as a negative control, as well as from samples obtained from 49 individuals collected in Magangué (Magdalena River drainage) and 24 individuals collected in Lorica (Sinú River drainage) in northern Colombia. Negative control individuals exhibited a baseline frequency of micronuclei of 0.78±0.58 and baseline values for comet tail length and tail moment of 3.34±0.24µm and 10.70±5.5, respectively. In contrast, samples from both field sites exhibited significantly greater evidence of genotoxic effects for both tests. The mean MN frequencies in the samples from Magangué and Lorica were 8.04±7.08 and 12.19±12.94, respectively. The mean tail length for samples from Magangué and Lorica were 5.78±3.18 and 15.46±7.39, respectively. Finally, the mean tail moment for samples from Magangué and

  19. Effects of β-glucan polysaccharide revealed by the dominant lethal assay and micronucleus assays, and reproductive performance of male mice exposed to cyclophosphamide

    Directory of Open Access Journals (Sweden)

    Rodrigo Juliano Oliveira

    2014-01-01

    Full Text Available β-glucan is a well-known polysaccharide for its chemopreventive effect. This study aimed to evaluate the chemopreventive ability of β-glucan in somatic and germ cells through the dominant lethal and micronucleus assays, and its influence on the reproductive performance of male mice exposed to cyclophosphamide. The results indicate that β-glucan is capable of preventing changes in DNA in both germ cells and somatic ones. Changes in germ cells were evaluated by the dominant lethal assay and showed damage reduction percentages of 46.46% and 43.79% for the doses of 100 and 150 mg/kg. For the somatic changes, evaluated by micronucleus assay in peripheral blood cells in the first week of treatment, damage reduction percentages from 80.63-116.32% were found. In the fifth and sixth weeks, the percentage ranged from 10.20-52.54% and -0.95-62.35%, respectively. Besides the chemopreventive efficiency it appears that the β-glucan, when combined with cyclophosphamide, is able to improve the reproductive performance of males verified by the significant reduction in rates of post-implantation losses and reabsorption in the mating of nulliparous females with males treated with cyclophosphamide.

  20. Lack of genotoxicity of potassium iodate in the alkaline comet assay and in the cytokinesis-block micronucleus test. Comparison to potassium bromate.

    Science.gov (United States)

    Poul, J M; Huet, S; Godard, T; Sanders, P

    2004-02-01

    Iodine could be added to the diet of human population in the form of iodide or iodate but iodate had not been adequately tested for genotoxicity and carcinogenicity. In the present study, genotoxic effects of potassium iodate were evaluated in vitro using the alkaline comet assay and the cytokinesis-block micronucleus assay on CHO cells and compared to halogenate salt analogues potassium bromate and chlorate and also to their respective reduced forms (potassium iodide, bromide and chloride). The results showed that the comet assay failed to detect the presence of DNA damage after a treatment of cells by potassium iodate for concentrations up to 10 mM. This absence of primary DNA damage was confirmed in the cytokinesis-block micronucleus assay. In the same way, results showed that potassium chlorate as well as potassium iodide, bromide and chloride did not induced DNA damage in the alkaline comet assay for doses up to 10 mM. By contrast, potassium bromate exposure led to an increase in both DNA damage and frequency of micronucleated cells. The repair of bromate-induced DNA damage was incomplete 24 h after the end of treatment. These results seem to indicate that potassium bromate would induce DNA damage by several mechanisms besides oxidative stress.

  1. Changes in buccal micronucleus cytome parameters associated with smokeless tobacco and pesticide exposure among female tea garden workers of Assam, India.

    Science.gov (United States)

    Kausar, Afifa; Giri, Sarbani; Roy, Prasenjit; Giri, Anirudha

    2014-03-01

    Assam is the highest tea producing state in India. A large number of workers are engaged in various units of tea industry. There are few reports on the health status of the tea garden workers. The present cytogenetic biomonitoring study was undertaken to investigate the genotoxic effect associated with workers in tea industries in southern Assam. Smokeless tobacco chewing along with betel nut is very common practice among the workers. Workers also get exposed periodically to mixture of pesticides. Employing buccal micronucleus cytome assay, exfoliated buccal cells were analyzed in 90 female tea garden and compared to 90 age and sex matched non-chewer control as well as 70 chewers who are not tea garden workers. Statistically significant (pworkers compared to both the control groups. The frequency of cell proliferation biomarkers was highest in the chewer controls whereas genotoxic and cell death parameters were highest in tea garden workers. Linear correlation analysis revealed strong positive correlation between the duration of occupation and the frequency of micronucleus (r=0.597; pworkers was relatively lower compared to the control group. Pesticide exposure and chewing areca nut along with smokeless tobacco use may be responsible for changes in cytome parameters in exfoliated buccal cells. Copyright © 2013 Elsevier GmbH. All rights reserved.

  2. Evaluation of repeated dose micronucleus assays of the liver using N-nitrosopyrrolidine: a report of the collaborative study by CSGMT/JEMS.MMS.

    Science.gov (United States)

    Ogawa, Izumi; Hagioa, Soichiro; Furukawa, Satoshi; Abe, Masayoshi; Kuroda, Yusuke; Hayashi, Seigo; Wako, Yumi; Kawasako, Kazufumi

    2015-03-01

    The repeated dose liver micronucleus (RDLMN) assay has the potential to detect liver carcinogens, and can be integrated into a general toxicological study. To assess the performance of the assay, N-nitrosopyrrolidine (NPYR), a genotoxic hepatocarcinogen, was tested in 14- or 28-day RDLMN assays. NPYR was orally administered to rats at a daily dose of 25, 50 or 100 mg/kg. One day after the last administration, a portion of the liver was removed and hepatocyte micronucleus (MN) specimens were prepared by the new method recently established by Narumi et al. In addition, a bone marrow MN assay and a histopathological examination of the liver were conducted. The detection of Phospho-Histone H3 was performed by immunohistochemistry to evaluate the proliferation rate of hepatocytes. The results showed significant increase in the number of micronucleated hepatocytes and Phospho-Histone H3-positive cells from the lowest dose in both 14- and 28-day RDLMN assays. On the other hand, the bone marrow MN assay yielded a negative result, which was in accordance with the existing report of the bone marrow MN assay using mice. Upon histopathological examination, inflammatory lesions and hypertrophy were noted, which may explain the increase in the hepatocyte proliferation and the enhancement of MN induction by NPYR. Our findings indicate that the RDLMN assay could be a useful tool for comprehensive risk assessment of carcinogenicity by providing information on both genotoxicity and histopathology when integrated into a general repeat dosing toxicity assay.

  3. Lack of genotoxic effect of food dyes amaranth, sunset yellow and tartrazine and their metabolites in the gut micronucleus assay in mice.

    Science.gov (United States)

    Poul, Martine; Jarry, Gérard; Elhkim, Mostafa Ould; Poul, Jean-Michel

    2009-02-01

    The food dyes amaranth, sunset yellow and tartrazine were administered twice, at 24h intervals, by oral gavage to mice and assessed in the in vivo gut micronucleus test for genotoxic effects (frequency of micronucleated cells) and toxicity (apoptotic and mitotic cells). The concentrations of each compound and their main metabolites (sulfanilic acid and naphthionic acid) were measured in faeces during a 24-h period after single oral administrations of the food dyes to mice. Parent dye compounds and their main aromatic amine metabolites were detected in significant amounts in the environment of colonic cells. Acute oral exposure to food dye additives amaranth, sunset yellow and tartrazine did not induce genotoxic effect in the micronucleus gut assay in mice at doses up to 2000 mg/kg b.w. Food dyes administration increased the mitotic cells at all dose levels when compared to controls. These results suggest that the transient DNA damages previously observed in the colon of mice treated by amaranth and tartrazine by the in vivo comet assay [Sasaki, Y.F., Kawaguchi, S., Kamaya, A., Ohshita, M., Kabasawa, K., Iwama, K., Taniguchi, K., Tsuda, S., 2002. The comet assay with 8 mouse organs: results with 39 currently used food additives. Mutat. Res. 519, 103-119] are unable to be fixed in stable genotoxic lesions and might be partly explained by local cytotoxicity of the dyes.

  4. Credit scoring methods

    Czech Academy of Sciences Publication Activity Database

    Vojtek, Martin; Kočenda, Evžen

    2006-01-01

    Roč. 56, 3-4 (2006), s. 152-167 ISSN 0015-1920 R&D Projects: GA ČR GA402/05/0931 Institutional research plan: CEZ:AV0Z70850503 Keywords : banking sector * credit scoring * discrimination analysis Subject RIV: AH - Economics Impact factor: 0.190, year: 2006 http://journal.fsv.cuni.cz/storage/1050_s_152_167.pdf

  5. Credit scoring for individuals

    Directory of Open Access Journals (Sweden)

    Maria DIMITRIU

    2010-12-01

    Full Text Available Lending money to different borrowers is profitable, but risky. The profits come from the interest rate and the fees earned on the loans. Banks do not want to make loans to borrowers who cannot repay them. Even if the banks do not intend to make bad loans, over time, some of them can become bad. For instance, as a result of the recent financial crisis, the capability of many borrowers to repay their loans were affected, many of them being on default. That’s why is important for the bank to monitor the loans. The purpose of this paper is to focus on credit scoring main issues. As a consequence of this, we presented in this paper the scoring model of an important Romanian Bank. Based on this credit scoring model and taking into account the last lending requirements of the National Bank of Romania, we developed an assessment tool, in Excel, for retail loans which is presented in the case study.

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

    Science.gov (United States)

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

    2018-01-01

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

  7. 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.

  8. 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.

  9. 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

  10. 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

  11. 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.

  12. 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

  13. 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.

  14. 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.

  15. College Math Assessment: SAT Scores vs. College Math Placement Scores

    Science.gov (United States)

    Foley-Peres, Kathleen; Poirier, Dawn

    2008-01-01

    Many colleges and university's use SAT math scores or math placement tests to place students in the appropriate math course. This study compares the use of math placement scores and SAT scores for 188 freshman students. The student's grades and faculty observations were analyzed to determine if the SAT scores and/or college math assessment scores…

  16. Estimating NHL Scoring Rates

    OpenAIRE

    Buttrey, Samuel E.; Washburn, Alan R.; Price, Wilson L.; Operations Research

    2011-01-01

    The article of record as published may be located at http://dx.doi.org/10.2202/1559-0410.1334 We propose a model to estimate the rates at which NHL teams score and yield goals. In the model, goals occur as if from a Poisson process whose rate depends on the two teams playing, the home-ice advantage, and the manpower (power-play, short-handed) situation. Data on all the games from the 2008-2009 season was downloaded and processed into a form suitable for the analysis. The model...

  17. The International Bleeding Risk Score

    DEFF Research Database (Denmark)

    Laursen, Stig Borbjerg; Laine, L.; Dalton, H.

    2017-01-01

    The International Bleeding Risk Score: A New Risk Score that can Accurately Predict Mortality in Patients with Upper GI-Bleeding.......The International Bleeding Risk Score: A New Risk Score that can Accurately Predict Mortality in Patients with Upper GI-Bleeding....

  18. 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.

  19. 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.

  20. 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...

  1. A follow-up survey of the lymphocyte micronucleus for three victims seventheen years after Shanghai '6.25' 60Co radiation accident

    International Nuclear Information System (INIS)

    Du Jie; Chen Ying; Zhang Xueqing; Liu Xiulin; Yan Xuekun

    2008-01-01

    Objective: To study the long-term effect on lymphocyte micronucleus and its clinical significance for the three victims seventheen years after Shanghai '6.25' 60 Co radiation accident. Method: The MN and MNC frequency were observed by CBMN method. Results: The MN and MNC frequency were still higher than normal even 17 years after the expo- sure. The MN frequency is related with the former radiation dose. The MNC consisting of ≥2MN is the hightest for 'Long', but for 'Jun' it consisted of 1 MN. The MN frequency is lower 17 years after the exposure than that of 5 years, but higher than 10 years. Conclusions: The MN is one of important signs for long-term radiation effect. Special attention should be payed to the high level of MN remaining in the body for so long time. (authors)

  2. Protection of nucleated bone marrow cells of mice against effect of radiation-induced micronucleus formation by using polysaccharides extracted from 'Zi Zhi' (a ganoderma)

    International Nuclear Information System (INIS)

    Chu Fang; Luo Houliang; Luo Gui; Chen Shunle; Liu Zhifang

    1988-01-01

    This paper describes the influence of polysaccharides extracted from 'Zi Zhi' (Ganoderma Sinese Zhao, Xu et Zhang) on the frequency of micronucleated cells induced by 60 Co gamma irradiation at different doses in bone marrow of mice. These polysaccharides of 'Zi Zhi' were shown to be of ability to protect nucleated bone marrow cells from micronucleus formation in irradiated mice. For Swiss mice, a dose reduction factor (DRF) was found to be 1.72 in the range of 0 to 4.728 Gy and for LACA mice, to be 1.73 in the range of 0 to 3.152 Gy. Such findings indicate that these polysaccharides are comparable to L-cysteine in their effeciency of protection

  3. Development of a data-processing method based on Bayesian k-means clustering to discriminate aneugens and clastogens in a high-content micronucleus assay.

    Science.gov (United States)

    Huang, Z H; Li, N; Rao, K F; Liu, C T; Huang, Y; Ma, M; Wang, Z J

    2018-03-01

    Genotoxicants can be identified as aneugens and clastogens through a micronucleus (MN) assay. The current high-content screening-based MN assays usually discriminate an aneugen from a clastogen based on only one parameter, such as the MN size, intensity, or morphology, which yields low accuracies (70-84%) because each of these parameters may contribute to the results. Therefore, the development of an algorithm that can synthesize high-dimensionality data to attain comparative results is important. To improve the automation and accuracy of detection using the current parameter-based mode of action (MoA), the MN MoA signatures of 20 chemicals were systematically recruited in this study to develop an algorithm. The results of the algorithm showed very good agreement (93.58%) between the prediction and reality, indicating that the proposed algorithm is a validated analytical platform for the rapid and objective acquisition of genotoxic MoA messages.

  4. Biomonitoring air quality during and after a public transportation strike in the center of Uberlândia, Minas Gerais, Brazil by Tradescantia micronucleus bioassay.

    Science.gov (United States)

    Pereira, Boscolli Barbosa; de Campos, Edimar Olegário; de Lima, Euclides Antônio Pereira; Barrozo, Marcos Antonio Souza; Morelli, Sandra

    2014-03-01

    The aim of this study was to address the lack of information concerning the air quality in the city of Uberlândia, Minas Gerais, Brazil. In this study, we conducted an unprecedented experiment involving the in situ biomonitoring of air genotoxicity in the city center during and after a public transportation strike using the Tradescantia micronucleus test. The frequency of micronuclei was significantly higher in the city center compared with the reference site (Mann-Whitney test, p transport stoppage (Kruskal-Wallis, Dunn p transport stoppage and the increase in the concentration of particulate matter from the increased flow of vehicles in the city center during the strike positively influenced the MN frequency. The climatic factors did not change during the biomonitoring period, reflecting the fact that climatic factors did not influence the MN frequency.

  5. The assessment of micronucleus frequency in lymphocytes in the cohort of coal-miners characterized by different polymorphisms of double strand break reparation genes

    Directory of Open Access Journals (Sweden)

    Maxim Yur'yevich Sinitsky

    2015-12-01

    Full Text Available Background: Coal-miners are exposed to a lot of number of harmful factors (chemical agents, ionizing radiation, heavy metals, coal dust etc.. Material and methods: Venous blood samples extracted from 129 coal-miners. Assessment of cytogenetic damage was performed using the cytokinesis-block micronucleus assay (CBMN on peripheral blood lymphocytes. PCR and gel electrophoresis were used to determine polymorphisms in the genes Lig4 (rs1805388 and XRCC4 (rs6869366. Results: We found a significant increase in the frequency of binucleated lymphocytes with micronuclei (MN and protrusions in carriers of the Ile/ Ile genotype of the Lig4 gene Thr9Ile polymorphism in comparison to Thr/Thr and Thr/Ile genotypes. Conclusions: Thr9Ile polymorphism within Lig4 gene can be used as potential molecular genetic markers of increased individual susceptibility to the complex of harmful factors in coal-mining conditions.

  6. Test of micronucleus in lymphocytes with the cytokinesis-block like possible indicator of the answer of the patient to the radiotherapy

    International Nuclear Information System (INIS)

    Giorgio, Marina di; Nasazzi, Nora; Taja, Maria R.; Roth, Berta; Sardi, Mabel; Menendez, Pablo R.

    2001-01-01

    In order to evaluate the individual cytogenetic response to radiotherapy and its comparison with the clinical response, the cytokinesis-block micronucleus assay was applied to peripheral blood lymphocytes of patients with cervix cancer undergoing radiotherapy. The cytogenetic data were analysed using a mathematical model to evaluate the attenuation of the cytogenetic effect as a function of the time between a single exposure and blood sampling, estimating a cytogenetic recovery factor (k) that might correlate with the individual radiosensitivity, contributing with radiosensitivity tests of current use but applying a rapid methodology easy to implement in a routine clinical laboratory. Long term clinical observations could confirm the validity of k in expressing predisposition of the subject to develop delayed effects. (author)

  7. Alkaline comet assay in liver and stomach, and micronucleus assay in bone marrow, from rats treated with 2-acetylaminofluorene, azidothymidine, cisplatin, or isobutyraldehyde.

    Science.gov (United States)

    Kraynak, A R; Barnum, J E; Cunningham, C L; Ng, A; Ykoruk, B A; Bennet, B; Stoffregen, D; Merschman, M; Freeland, E; Galloway, S M

    2015-07-01

    As part of the Japanese Center for the Validation of Alternative Methods (JaCVAM) initiative international validation study of the in vivo rat alkaline comet assay (comet assay), we examined the ability of the assay to determine the genotoxicity of 2-acetylaminofluorene (AAF), azidothymidine (AZT), cisplatin (CPN), and isobutyraldehyde (IBA) in liver and glandular stomach of male Sprague-Dawley rats. Rats were given oral doses of test compound or control once daily for three days. High dose levels were approximately maximum tolerated doses and were based on preliminary range-finding studies. Tissues were harvested 3h after the final dose (48h after the initial dose). A bone marrow micronucleus assay (MN) was also conducted on the rats treated with AZT, CPN, and IBA. Acute toxic effects of treatment were determined primarily through histomorphologic analysis of liver and stomach but also by body weight and serum liver enzyme changes. The comet assay was conducted on fresh tissue preparations but frozen samples from two studies were also assayed. Statistically significant dose-related differences in comet % DNA in tail were found in liver and stomach for the genotoxin AZT and in liver for the genotoxin CPN, but not in liver or stomach for the non-genotoxin IBA. Statistically significant differences in % DNA in tail were measured in liver for the low and mid dose of the genotoxin AAF, but not the high dose. The comet assays of frozen liver suspensions from CPN- and AAF-treated rats yielded comparable results to the assays of fresh preparations. There were no indications of significant toxicity induced by any treatment. The micronucleus assay was positive for CPN and AZT and negative for IBA. In conclusion, the in vivo comet assay is capable of detecting genotoxic effects of a variety of chemicals and may fill an important role in the genotoxicity test battery. Copyright © 2015 Elsevier B.V. All rights reserved.

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

    Science.gov (United States)

    Ondigo, Bartholomew N; Park, Gregory S; Gose, Severin O; Ho, Benjamin M; Ochola, Lyticia A; Ayodo, George O; Ofulla, Ayub V; John, Chandy C

    2012-12-21

    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. 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. 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 <0.0001 - 0.004), and antibodies to variants of the same antigen were accurately distinguished within a multiplex reaction. Plasma dilutions of 1:100 or 1:200 were optimal for all antigens for CBA testing. Plasma diluted in a buffer containing 0.05% sodium azide, 0.5% polyvinylalcohol, and 0.8% polyvinylpyrrolidone had the lowest background activity. CBA median fluorescence intensity (MFI) values with 1,000 antigen-conjugated beads/well did not differ significantly from MFI with 5,000 beads/well. CBA and ELISA results correlated well for all antigens except apical membrane antigen-1 (AMA-1). CBA testing produced a greater range of values in samples from malaria endemic areas and less background reactivity for blank samples than ELISA. 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

  9. Do Test Scores Buy Happiness?

    Science.gov (United States)

    McCluskey, Neal

    2017-01-01

    Since at least the enactment of No Child Left Behind in 2002, standardized test scores have served as the primary measures of public school effectiveness. Yet, such scores fail to measure the ultimate goal of education: maximizing happiness. This exploratory analysis assesses nation level associations between test scores and happiness, controlling…

  10. Predicting occupational personality test scores.

    Science.gov (United States)

    Furnham, A; Drakeley, R

    2000-01-01

    The relationship between students' actual test scores and their self-estimated scores on the Hogan Personality Inventory (HPI; R. Hogan & J. Hogan, 1992), an omnibus personality questionnaire, was examined. Despite being given descriptive statistics and explanations of each of the dimensions measured, the students tended to overestimate their scores; yet all correlations between actual and estimated scores were positive and significant. Correlations between self-estimates and actual test scores were highest for sociability, ambition, and adjustment (r = .62 to r = .67). The results are discussed in terms of employers' use and abuse of personality assessment for job recruitment.

  11. Differences of wells scores accuracy, caprini scores and padua scores in deep vein thrombosis diagnosis

    Science.gov (United States)

    Gatot, D.; Mardia, A. I.

    2018-03-01

    Deep Vein Thrombosis (DVT) is the venous thrombus in lower limbs. Diagnosis is by using venography or ultrasound compression. However, these examinations are not available yet in some health facilities. Therefore many scoring systems are developed for the diagnosis of DVT. The scoring method is practical and safe to use in addition to efficacy, and effectiveness in terms of treatment and costs. The existing scoring systems are wells, caprini and padua score. There have been many studies comparing the accuracy of this score but not in Medan. Therefore, we are interested in comparative research of wells, capriniand padua score in Medan.An observational, analytical, case-control study was conducted to perform diagnostic tests on the wells, caprini and padua score to predict the risk of DVT. The study was at H. Adam Malik Hospital in Medan.From a total of 72 subjects, 39 people (54.2%) are men and the mean age are 53.14 years. Wells score, caprini score and padua score has a sensitivity of 80.6%; 61.1%, 50% respectively; specificity of 80.65; 66.7%; 75% respectively, and accuracy of 87.5%; 64.3%; 65.7% respectively.Wells score has better sensitivity, specificity and accuracy than caprini and padua score in diagnosing DVT.

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

    Science.gov (United States)

    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

  13. [Propensity score matching in SPSS].

    Science.gov (United States)

    Huang, Fuqiang; DU, Chunlin; Sun, Menghui; Ning, Bing; Luo, Ying; An, Shengli

    2015-11-01

    To realize propensity score matching in PS Matching module of SPSS and interpret the analysis results. The R software and plug-in that could link with the corresponding versions of SPSS and propensity score matching package were installed. A PS matching module was added in the SPSS interface, and its use was demonstrated with test data. Score estimation and nearest neighbor matching was achieved with the PS matching module, and the results of qualitative and quantitative statistical description and evaluation were presented in the form of a graph matching. Propensity score matching can be accomplished conveniently using SPSS software.

  14. [Prognostic scores for pulmonary embolism].

    Science.gov (United States)

    Junod, Alain

    2016-03-23

    Nine prognostic scores for pulmonary embolism (PE), based on retrospective and prospective studies, published between 2000 and 2014, have been analyzed and compared. Most of them aim at identifying PE cases with a low risk to validate their ambulatory care. Important differences in the considered outcomes: global mortality, PE-specific mortality, other complications, sizes of low risk groups, exist between these scores. The most popular score appears to be the PESI and its simplified version. Few good quality studies have tested the applicability of these scores to PE outpatient care, although this approach tends to already generalize in the medical practice.

  15. Genotoxic evaluation of [DOTA,Tyr3]octreotate labeled with 131I and 177Lu in human peripheral lymphocytes in vitro by micronucleus assay

    International Nuclear Information System (INIS)

    Suzauki, Miriam Fussae; Silva, Marcia Augusta da; Caldeira Filho, Jose de Souza; Colturato, Maria Tereza; Araujo, Elaine Bortoleti de; Bartolini, Paolo; Okazaki, Kayo

    2005-01-01

    The radiolabeled receptor-binding peptides have being used for cancer diagnosis and therapy. The octreotate, a somatostatin analogue peptide, bound to various tumors expressing sst receptors (thyroid, pancreas, prostrate, melanoma and lymphomas). The amount and the type of receptors for somatostatin influence the tissue uptake. The [DOTA, Tyr 3 ]octreotate has been used because of its high affinity to somatostatin subtype receptors sstr 2 and sstr 5 . The pharmacokinetic study showed that the blood clearance is rapid and only 9% of the intravenous injected activity remains in human blood after one hour. The aim of this study was to evaluate the cytogenetic effect of radiolabeled [DOTA, Tyr 3 ]octreotate in blood cells in vitro, using the cytokinesis-block micronucleus (MN) assay. This technique allows evaluating the mutagenic effects of both endogenous and exogenous agents at chromosome level. Blood samples of healthy donors were collected in heparinized syringes and exposed to different activities of [DOTA, Tyr 3 ]octreotate labeled with with 131 I (n=3) and 177 Lu (n=3), where radioactive concentration ranged from 600 to 5600 kBq/mL, corresponding to an injected activity of 3.1 to 28.9 GBq in a reference man of 70 kg weight. 131 I and 177 Lu are beta- and gamma-emitters. After one-hour exposition to radiopharmaceuticals at 37 deg C, the cells were washed with culture medium for removing the non internalised octreotate and cultivated for 72 hours, according to criteria adopted by the IAEA. The results showed a positive correlation between radioactive concentrations (X) and the frequency of binucleated cells with micronuclei (Y) (P 131 I-DOTA, Tyr 3 ]octreotate was Y = (1.634 ± 0.236) + (0.912 ± 0.137) 10 -3 X and for [ 177 Lu-DOTA, Tyr 3 ]octreotate was Y = (1.715 ± 0.342) + (0.743 ± 0.135) 10 -3 X. The non labeled molecule, [DOTA, Tyr 3 ]octreotate, has no influence in the induction of cytogenetic damage. The micronucleus assay with rat pancreatic tumor cells

  16. Cadmium chloride, benzo[a]pyrene and cyclophosphamide tested in the in vitro mammalian cell micronucleus test (MNvit) in the human lymphoblastoid cell line TK6 at Covance laboratories, Harrogate UK in support of OECD draft Test Guideline 487.

    Science.gov (United States)

    Fowler, Paul; Whitwell, James; Jeffrey, Laura; Young, Jamie; Smith, Katie; Kirkland, David

    2010-10-29

    The following genotoxic chemicals were tested in the in vitro micronucleus assay, at Covance Laboratories, Harrogate, UK in the human lymphoblastoid cell line TK6. Cadmium chloride (an inorganic carcinogen), benzo[a]pyrene (a polycyclic aromatic hydrocarbon requiring metabolic activation) and cyclophosphamide (an alkylating agent requiring metabolic activation) were treated with and without cytokinesis block (by addition of cytochalasin B). This work formed part of a collaborative evaluation of the toxicity measures recommended in the draft OECD Test Guideline 487 for the in vitro micronucleus test. The toxicity measures used, capable of detecting both cytostasis and cell death, were relative population doubling, relative increase in cell counts and relative cell counts for treatments in the absence of cytokinesis block, and replication index or cytokinesis blocked proliferation index in the presence of cytokinesis block. All of the chemicals tested gave significant increases in the percentage of micronucleated cells with and without cytokinesis block at concentrations giving approximately 60% toxicity (cytostasis and cell death) or less by all of the toxicity measures used. The outcomes from this series of tests support the use of relative increase in cell counts and relative population doubling, as well as relative cell counts, as appropriate measures of cytotoxicity for the non-cytokinesis blocked in the in vitro micronucleus assay. Copyright © 2010 Elsevier B.V. All rights reserved.

  17. Etoposide; colchicine; mitomycin C and cyclophosphamide tested in the in vitro mammalian cell micronucleus test (MNvit) in Chinese hamster lung (CHL) cells at Covance laboratories; Harrogate UK in support of OECD draft Test Guideline 487.

    Science.gov (United States)

    Fowler, Paul; Whitwell, James; Jeffrey, Laura; Young, Jamie; Smith, Katie; Kirkland, David

    2010-10-29

    The following genotoxic chemicals were tested in the in vitro micronucleus assay, at Covance Laboratories, Harrogate, UK in the Chinese hamster lung cell line CHL. Etoposide (a topoisomerase inhibitor), colchicine (an aneugen), mitomycin C (a DNA cross linking agent) and cyclophosphamide (an alkylating agent requiring metabolic activation) were treated with and without cytokinesis block (by addition of cytochalasin B). This work formed part of a collaborative evaluation of the toxicity measures recommended in the draft OECD Test Guideline 487 for the in vitro micronucleus test. The toxicity measures used, detecting both cytostasis and cell death, were relative population doubling, relative increase in cell counts and relative cell counts for treatments in the absence of cytokinesis block, and replication index or cytokinesis blocked proliferation index in the presence of cytokinesis block. All of the chemicals tested gave significant increases in the percentage of micronucleated cells with and without cytokinesis block at concentrations giving approximately 60% toxicity (cytostasis and cell death) or less by all of the toxicity measures used. The outcomes from this series of tests support the use of relative increase in cell counts and relative population doubling, as well as relative cell counts, as appropriate measures of cytotoxicity for the non-cytokinesis blocked in vitro micronucleus assay. Copyright © 2010 Elsevier B.V. All rights reserved.

  18. 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, ...

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

    Science.gov (United States)

    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.

  20. D-score: a search engine independent MD-score.

    Science.gov (United States)

    Vaudel, Marc; Breiter, Daniela; Beck, Florian; Rahnenführer, Jörg; Martens, Lennart; Zahedi, René P

    2013-03-01

    While peptides carrying PTMs are routinely identified in gel-free MS, the localization of the PTMs onto the peptide sequences remains challenging. Search engine scores of secondary peptide matches have been used in different approaches in order to infer the quality of site inference, by penalizing the localization whenever the search engine similarly scored two candidate peptides with different site assignments. In the present work, we show how the estimation of posterior error probabilities for peptide candidates allows the estimation of a PTM score called the D-score, for multiple search engine studies. We demonstrate the applicability of this score to three popular search engines: Mascot, OMSSA, and X!Tandem, and evaluate its performance using an already published high resolution data set of synthetic phosphopeptides. For those peptides with phosphorylation site inference uncertainty, the number of spectrum matches with correctly localized phosphorylation increased by up to 25.7% when compared to using Mascot alone, although the actual increase depended on the fragmentation method used. Since this method relies only on search engine scores, it can be readily applied to the scoring of the localization of virtually any modification at no additional experimental or in silico cost. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Trends in Classroom Observation Scores

    Science.gov (United States)

    Casabianca, Jodi M.; Lockwood, J. R.; McCaffrey, Daniel F.

    2015-01-01

    Observations and ratings of classroom teaching and interactions collected over time are susceptible to trends in both the quality of instruction and rater behavior. These trends have potential implications for inferences about teaching and for study design. We use scores on the Classroom Assessment Scoring System-Secondary (CLASS-S) protocol from…

  2. Quadratic prediction of factor scores

    NARCIS (Netherlands)

    Wansbeek, T

    1999-01-01

    Factor scores are naturally predicted by means of their conditional expectation given the indicators y. Under normality this expectation is linear in y but in general it is an unknown function of y. II is discussed that under nonnormality factor scores can be more precisely predicted by a quadratic

  3. The Machine Scoring of Writing

    Science.gov (United States)

    McCurry, Doug

    2010-01-01

    This article provides an introduction to the kind of computer software that is used to score student writing in some high stakes testing programs, and that is being promoted as a teaching and learning tool to schools. It sketches the state of play with machines for the scoring of writing, and describes how these machines work and what they do.…

  4. Matching score based face recognition

    NARCIS (Netherlands)

    Boom, B.J.; Beumer, G.M.; Spreeuwers, Lieuwe Jan; Veldhuis, Raymond N.J.

    2006-01-01

    Accurate face registration is of vital importance to the performance of a face recognition algorithm. We propose a new method: matching score based face registration, which searches for optimal alignment by maximizing the matching score output of a classifier as a function of the different

  5. Modelling sequentially scored item responses

    NARCIS (Netherlands)

    Akkermans, W.

    2000-01-01

    The sequential model can be used to describe the variable resulting from a sequential scoring process. In this paper two more item response models are investigated with respect to their suitability for sequential scoring: the partial credit model and the graded response model. The investigation is

  6. 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.

  7. 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.

  8. Evaluation of the repeated-dose liver and gastrointestinal tract micronucleus assays with 22 chemicals using young adult rats: summary of the collaborative study by the Collaborative Study Group for the Micronucleus Test (CSGMT)/The Japanese Environmental Mutagen Society (JEMS) - Mammalian Mutagenicity Study Group (MMS).

    Science.gov (United States)

    Hamada, Shuichi; Ohyama, Wakako; Takashima, Rie; Shimada, Keisuke; Matsumoto, Kazumi; Kawakami, Satoru; Uno, Fuyumi; Sui, Hajime; Shimada, Yasushi; Imamura, Tadashi; Matsumura, Shoji; Sanada, Hisakazu; Inoue, Kenji; Muto, Shigeharu; Ogawa, Izumi; Hayashi, Aya; Takayanagi, Tomomi; Ogiwara, Yosuke; Maeda, Akihisa; Okada, Emiko; Terashima, Yukari; Takasawa, Hironao; Narumi, Kazunori; Wako, Yumi; Kawasako, Kazufumi; Sano, Masaki; Ohashi, Nobuyuki; Morita, Takeshi; Kojima, Hajime; Honma, Masamitsu; Hayashi, Makoto

    2015-03-01

    The repeated-dose liver micronucleus (RDLMN) assay using young adult rats has the potential to detect hepatocarcinogens. We conducted a collaborative study to assess the performance of this assay and to evaluate the possibility of integrating it into general toxicological studies. Twenty-four testing laboratories belonging to the Mammalian Mutagenicity Study Group, a subgroup of the Japanese Environmental Mutagen Society, participated in this trial. Twenty-two model chemicals, including some hepatocarcinogens, were tested in 14- and/or 28-day RDLMN assays. As a result, 14 out of the 16 hepatocarcinogens were positive, including 9 genotoxic hepatocarcinogens, which were reported negative in the bone marrow/peripheral blood micronucleus (MN) assay by a single treatment. These outcomes show the high sensitivity of the RDLMN assay to hepatocarcinogens. Regarding the specificity, 4 out of the 6 non-liver targeted genotoxic carcinogens gave negative responses. This shows the high organ specificity of the RDLMN assay. In addition to the RDLMN assay, we simultaneously conducted gastrointestinal tract MN assays using 6 of the above carcinogens as an optional trial of the collaborative study. The MN assay using the glandular stomach, which is the first contact site of the test chemical when administered by oral gavage, was able to detect chromosomal aberrations with 3 test chemicals including a stomach-targeted carcinogen. The treatment regime was the 14- and/or 28-day repeated-dose, and the regime is sufficiently promising to incorporate these methods into repeated-dose toxicological studies. The outcomes of our collaborative study indicated that the new techniques to detect chromosomal aberrations in vivo in several tissues worked successfully. Copyright © 2015 The Authors. Published by Elsevier B.V. All rights reserved.

  9. Evaluation of the repeated-dose liver micronucleus assay using N-nitrosomorpholine in young adult rats: report on collaborative study by the Collaborative Study Group for the Micronucleus Test (CSGMT)/Japanese Environmental Mutagen Society (JEMS)-Mammalian Mutagenicity Study (MMS) Group.

    Science.gov (United States)

    Hayashi, Aya; Kosaka, Mizuki; Kimura, Aoi; Wako, Yumi; Kawasako, Kazufumi; Hamada, Shuichi

    2015-03-01

    The present study was conducted to evaluate the suitability of a repeated-dose liver micronucleus (LMN) assay in young adult rats as a collaborative study by the Mammalian mutagenicity study (MMS) group. All procedures were performed in accordance with the standard protocols of the MMS Group. Six-week-old male Crl:CD(SD) rats (5 animals/group) received oral doses of the hepatocarcinogen N-nitrosomorpholine (NMOR) at 0 (control), 5, 10, and 30mg/kg/day (10mL/kg) for 14 days. Control animals received vehicle (water). Hepatocytes were collected from the liver 24h after the last dose, and the number of micronucleated hepatocytes (MNHEPs) was determined by microscopy. The number of micronucleated immature erythrocytes (MNIMEs) in the femoral bone marrow was also determined. The liver was examined using histopathologic methods after formalin fixation. The results showed statistically significant and dose-dependent increases in the number of MNHEPs in the liver at doses of 10mg/kg and greater when compared with the vehicle control. However, no significant increase was noted in the number of MNIMEs in the bone marrow at doses of up to 30mg/kg. Histopathology of the liver revealed hypertrophy and single cell necrosis of hepatocytes at doses of 5mg/kg and above. These results showed that the induction of micronuclei by NMOR was detected by the repeated-dose LMN assay, but not by the repeated-dose bone marrow micronucleus assay. Copyright © 2014 Elsevier B.V. All rights reserved.

  10. Genotoxic and antigenotoxic properties of selenium compounds in the in vitro micronucleus assay with human whole blood lymphocytes and tk6 lymphoblastoid cells

    Directory of Open Access Journals (Sweden)

    Eduard Cemeli

    2006-01-01

    Full Text Available Selenium is known to possess both genotoxic and antigenotoxic properties. In the present study, we have evaluated the genotoxicity and antigenotoxicity of three selenium compounds (sodium selenate, sodium selenite and selenous acid by measuring in vitro micronucleus induction. Assays were conducted in whole blood lymphocytes and in the TK6 lymphoblastoid cell line, with and without co-treatment with potassium dichromate, a known genotoxic compound. In general, the compounds were more active in TK6 cells than they were in blood lymphocytes. Only 1 μM selenous acid increased the frequency of binucleated cells containing micronuclei (BNMN in blood lymphocytes, while all three selenium compounds increased BNMN in TK6 cells. In addition, combinations of selenous acid and potassium dichromate resulted in lower frequencies of BNMN than potassium dichromate alone in blood lymphocytes, while combinations of sodium selenate and potassium dichromate produced lower frequencies of BNMN than potassium dichromate alone in TK6 cells. The concentrations of selenium compounds that were used, in combination with the medium components and the biological physiology of the whole blood lymphocytes and TK6 cells, could have affected the redox potential of the compounds, switching the chemicals from a pro-oxidant to antioxidant status and vice-versa. The lower activities of the compounds in blood lymphocytes may be due to the protective effects of blood components. The results indicate that the genotoxic and antigenotoxic properties of selenium compounds are highly dependent upon the conditions under which they are evaluated.

  11. Hydrocortisone Increases the Vinblastine-Induced Chromosomal Damages in L929 Cells Investigated by the Micronucleus Assay on Cytokinesis-Blocked Binucleated Cells

    Directory of Open Access Journals (Sweden)

    Tahere Ebrahimipour

    2017-03-01

    Full Text Available Background: Stress may cause damages to DNA or/and change the ability of the cells to overcome these damages. It may also cause irregularities in the cell cycle and induce abnormal cell divisions through glucocorticoid-dependent functions. The abnormal cell divisions, in turn, lead to chromosomal mal-segregation and aneuploidy. In this study, the effects of the stress hormone, hydrocortisone (HYD, were investigated on the induced chromosomal abnormalities by vinblastine (VIN during cell cycle in L929 cells. Methods: This work was performed in winter 2013 at Department of Biology, University of Ferdowsi, Mashhad, Iran. Cultured cells were divided into different groups including control, VIN-treated, HYD treated and VIN+HYD co-treated cells. The induced chromosomal damages were investigated by micronucleus assay in cytokinesis-blocked binucleated cells. Results: Although HYD by itself did not increase the micronuclei (Mn frequency, co-treatment of cells with VIN and HYD led to significant increase (P<0.05 in the frequency of Mn in comparison to control and VIN treated groups. Conclusion: Cells treated with stress hormone are more sensitive to damages induced by VIN. Therefore, stress may not directly result in genetic instability, it can increase the harmful effects associated with other genotoxic agents.

  12. Evaluation of apoptosis and apoptosis proteins as possible markers of radiation at doses 0.1-2 Gy, in comparison to the micronucleus assay in three cell lines

    International Nuclear Information System (INIS)

    Jaworska, A.; Angelis, P. de

    1997-01-01

    In recent years the interest in apoptosis as possible indicator of radiation damage has increased. Studies have been done to examine the induction of apoptosis after ionizing radiation using morphological criteria, characteristic DNA damage pattern(ladders), early DNA damage using flow cytometry and/or expression of the proteins involved in apoptosis. But the picture which emerges from these investigations is unclear. Some researchers suggest that apoptosis studies can be used as potential assays of biological dosimetry, others doubt if apoptosis can be used as a marker of irradiation at all. Most studies have been done using relatively high doses of radiation. In this study we focus on apoptosis induction after relatively small doses (0,1-2 Gy). We detected apoptosis with the in situ terminal deoxynucleotidyl transferase assay by flow cytometry, and measured the expression of proteins that regulate apoptosis (Bcl-2, Bax, P53) with Western blotting. As comparison we used the cytokinesis-block micronucleus assay as a reference. The studies were carried out in three lymphoid cell lines: the mouse lymphoma L5178Y resistant and sensitive cell lines widely used in radiobiological studies, and the human pre-B cell leukemia Reh cells. Our results indicate that we can not consider the examined parameters of apoptosis as markers of radiation in these cell lines. (author)

  13. Genotoxic and Antigenotoxic Assessment of Chios Mastic Oil by the In Vitro Micronucleus Test on Human Lymphocytes and the In Vivo Wing Somatic Test on Drosophila.

    Directory of Open Access Journals (Sweden)

    Dimitris Vlastos

    Full Text Available Chios mastic oil (CMO, the essential oil derived from Pistacia lentiscus (L. var. chia (Duham, has generated considerable interest because of its antimicrobial, anticancer, antioxidant and other beneficial properties. In the present study, the potential genotoxic activity of CMO as well as its antigenotoxic properties against the mutagenic agent mitomycin-C (MMC were evaluated by employing the in vitro Cytokinesis Block MicroNucleus (CBMN assay and the in vivo Somatic Mutation And Recombination Test (SMART. In the in vitro experiments, lymphocytes were treated with 0.01, 0.05 and 0.10% (v/v of CMO with or without 0.05 μg/ml MMC, while in the in vivo assay Drosophila larvae were fed with 0.05, 0.10, 0.50 and 1.00% (v/v of CMO with or without 2.50 μg/ml MMC. CMO did not significantly increase the frequency of micronuclei (MN or total wing spots, indicating lack of mutagenic or recombinogenic activity. However, the in vitro analysis suggested cytotoxic activity of CMO. The simultaneous administration of MMC with CMO did not alter considerably the frequencies of MMC-induced MN and wing spots showing that CMO doesn't exert antigenotoxic or antirecombinogenic action. Therefore, CMO could be considered as a safe product in terms of genotoxic potential. Even though it could not afford any protection against DNA damage, at least under our experimental conditions, its cytotoxic potential could be of interest.

  14. Enhanced micronucleus formation in the descendants of {gamma}-ray-irradiated tobacco cells: Evidence for radiation-induced genomic instability in plant cells

    Energy Technology Data Exchange (ETDEWEB)

    Yokota, Yuichiro, E-mail: yokota.yuichiro@jaea.go.jp [Life Science and Biotechnology Division, Quantum Beam Science Directorate, Japan Atomic Energy Agency, 1233 Watanuki-machi, Takasaki, Gunma 370-1292 (Japan); Funayama, Tomoo; Hase, Yoshihiro [Life Science and Biotechnology Division, Quantum Beam Science Directorate, Japan Atomic Energy Agency, 1233 Watanuki-machi, Takasaki, Gunma 370-1292 (Japan); Hamada, Nobuyuki [Radiation Safety Research Center, Nuclear Technology Research Laboratory, Central Research Institute of Electric Power Industry, 2-11-1 Iwado-kita, Komae, Tokyo 201-8511 (Japan); Kobayashi, Yasuhiko; Tanaka, Atsushi; Narumi, Issay [Life Science and Biotechnology Division, Quantum Beam Science Directorate, Japan Atomic Energy Agency, 1233 Watanuki-machi, Takasaki, Gunma 370-1292 (Japan)

    2010-09-10

    Ionizing radiation-induced genomic instability has been documented in various end points such as chromosomal aberrations and mutations, which arises in the descendants of irradiated mammalian or yeast cells many generations after the initial insult. This study aimed at addressing radiation-induced genomic instability in higher plant tobacco cells. We thus investigated micronucleus (MN) formation and cell proliferation in tobacco cells irradiated with {gamma}-rays and their descendants. In {gamma}-irradiated cells, cell cycle was arrested at G{sub 2}/M phase at around 24 h post-irradiation but released afterward. In contrast, MN frequency peaked at 48 h post-irradiation. Almost half of 40 Gy-irradiated cells had MN at 48 h post-irradiation, but proliferated as actively as sham-irradiated cells up to 120 h post-irradiation. Moreover, the descendants that have undergone at least 22 generations after irradiation still showed a two-fold MN frequency compared to sham-irradiated cells. This is the direct evidence for radiation-induced genomic instability in tobacco cells.

  15. Vinclozolin, a widely used fungizide, enhanced BaP-induced micronucleus formation in human derived hepatoma cells by increasing CYP1A1 expression.

    Science.gov (United States)

    Wu, Xin-Jiang; Lu, Wen-qing; Roos, Peter H; Mersch-Sundermann, Volker

    2005-10-15

    Vinclozolin, a widely used fungicide, can be identified as a residue in numerous vegetable and fruit samples. To get insight in its genetic toxicity, we investigated the genotoxic effect of vinclozolin in the human derived hepatoma cell line HepG2 using the micronucleus (MN) assay. Additionally, to evaluate the co- or anti-mutagenic potency of vinclozolin, we treated HepG2 cells with different concentrations of vinclozolin for 24 h. Subsequently, the cells were exposed to benzo[a]pyrene (BaP) for 1h. Exposure of HepG2 cells to 50-400 microM vinclozolin alone did not cause any induction of micronuclei. However, a pronounced co-mutagenic effect was observed. MN frequencies caused by BaP increased by 30.6%, 52.8% and 65.3% after pretreatment of the cell cultures with 50, 100 and 200 microM vinclozolin, respectively. The highest concentration (400 microM) of vinclozolin tested caused cytotoxicity. Therefore, micronuclei were not considered for that concentration. To clarify the mechanism of cogenotoxicity, we assayed cytochrome P450 1A1 (CYP1A1), which plays a pivotal role in activation of BaP. Cells exposed to vinclozolin led to significant increase of CYP1A1 expression in Western blot. The result suggested that induction of CYP1A1 by vinclozolin account for its enhancing effect on genotoxicity caused by BaP.

  16. Evaluation of repeated dose micronucleus assays of the liver and gastrointestinal tract using potassium bromate: a report of the collaborative study by CSGMT/JEMS.MMS.

    Science.gov (United States)

    Okada, Emiko; Fujiishi, Yohei; Narumi, Kazunori; Kado, Shoichi; Wako, Yumi; Kawasako, Kazufumi; Kaneko, Kimiyuki; Ohyama, Wakako

    2015-03-01

    The food additive potassium bromate (KBrO3) is known as a renal carcinogen and causes chromosomal aberrations in vitro without metabolic activation and in vivo in hematopoietic and renal cells. As a part of a collaborative study by the Mammalian Mutagenicity Study group, which is a subgroup of the Japanese Environmental Mutagen Society, we administered KBrO3 to rats orally for 4, 14, and 28 days and examined the micronucleated (MNed) cell frequency in the liver, glandular stomach, colon, and bone marrow to confirm whether the genotoxic carcinogen targeting other than liver and gastrointestinal (GI) tract was detected by the repeated dose liver and GI tract micronucleus (MN) assays. In our study, animals treated with KBrO3 showed some signs of toxicity in the kidney and/or stomach. KBrO3 did not increase the frequency of MNed cells in the liver and colon in any of the repeated dose studies. However, KBrO3 increased the frequency of MNed cells in the glandular stomach and bone marrow. Additionally, the MNed cell frequency in the glandular stomach was not significantly affected by the difference in the length of the administration period. These results suggest that performing the MN assay using the glandular stomach, which is the first tissue to contact agents after oral ingestion, is useful for evaluating the genotoxic potential of chemicals and that the glandular stomach MN assay could be integrated into general toxicity studies. Copyright © 2014 Elsevier B.V. All rights reserved.

  17. Evaluation of the repeated dose liver micronucleus assay using young adult rats with cyclophosphamide monohydrate: a report of a collaborative study by CSGMT/JEMS.MMS.

    Science.gov (United States)

    Matsumoto, Kazumi; Zaizen, Kazuyo; Miyamoto, Atsushi; Wako, Yumi; Kawasako, Kazufumi; Ishida, Hisao

    2015-03-01

    The repeated dose liver micronucleus (RDLMN) assay using young adult rats has the potential to detect liver carcinogens, and this assay could be integrated into general toxicological studies. In this study, in order to assess the performance of the assay, cyclophosphamide monohydrate (CP) was tested in a 14-day RDLMN assay. Based on the results of the 4-day repeated dose-finding study, 10 mg/kg/day of CP was selected as the highest dose and the lower doses were set at 5, 2.5, 1.25, and 0.625 mg/kg/day for the 14-day RDLMN assay. On the day after the completion of the dosing period, specimens of hepatocytes and bone marrow cells were prepared and the induction of micronuclei was assessed. No changes were observed in the incidences of micronucleated hepatocytes. Nevertheless, the incidences of micronucleated immature erythrocytes in the bone marrow were increased significantly at CP doses of 1.25 mg/kg/day or more. These findings are consistent with reports that CP induces tumors in various tissues but it does not induce liver tumors.

  18. Extension of the lod score: the mod score.

    Science.gov (United States)

    Clerget-Darpoux, F

    2001-01-01

    In 1955 Morton proposed the lod score method both for testing linkage between loci and for estimating the recombination fraction between them. If a disease is controlled by a gene at one of these loci, the lod score computation requires the prior specification of an underlying model that assigns the probabilities of genotypes from the observed phenotypes. To address the case of linkage studies for diseases with unknown mode of inheritance, we suggested (Clerget-Darpoux et al., 1986) extending the lod score function to a so-called mod score function. In this function, the variables are both the recombination fraction and the disease model parameters. Maximizing the mod score function over all these parameters amounts to maximizing the probability of marker data conditional on the disease status. Under the absence of linkage, the mod score conforms to a chi-square distribution, with extra degrees of freedom in comparison to the lod score function (MacLean et al., 1993). The mod score is asymptotically maximum for the true disease model (Clerget-Darpoux and Bonaïti-Pellié, 1992; Hodge and Elston, 1994). Consequently, the power to detect linkage through mod score will be highest when the space of models where the maximization is performed includes the true model. On the other hand, one must avoid overparametrization of the model space. For example, when the approach is applied to affected sibpairs, only two constrained disease model parameters should be used (Knapp et al., 1994) for the mod score maximization. It is also important to emphasize the existence of a strong correlation between the disease gene location and the disease model. Consequently, there is poor resolution of the location of the susceptibility locus when the disease model at this locus is unknown. Of course, this is true regardless of the statistics used. The mod score may also be applied in a candidate gene strategy to model the potential effect of this gene in the disease. Since, however, it

  19. 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.

  20. From Rasch scores to regression

    DEFF Research Database (Denmark)

    Christensen, Karl Bang

    2006-01-01

    Rasch models provide a framework for measurement and modelling latent variables. Having measured a latent variable in a population a comparison of groups will often be of interest. For this purpose the use of observed raw scores will often be inadequate because these lack interval scale propertie....... This paper compares two approaches to group comparison: linear regression models using estimated person locations as outcome variables and latent regression models based on the distribution of the score....

  1. 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...

  2. Re-Scoring the Game’s Score

    DEFF Research Database (Denmark)

    Gasselseder, Hans-Peter

    2014-01-01

    This study explores immersive presence as well as emotional valence and arousal in the context of dynamic and non-dynamic music scores in the 3rd person action-adventure video game genre while also considering relevant personality traits of the player. 60 subjects answered self-report questionnai......This study explores immersive presence as well as emotional valence and arousal in the context of dynamic and non-dynamic music scores in the 3rd person action-adventure video game genre while also considering relevant personality traits of the player. 60 subjects answered self......-temporal alignment in the resulting emotional congruency of nondiegetic music. Whereas imaginary aspects of immersive presence are systemically affected by the presentation of dynamic music, sensory spatial aspects show higher sensitivity towards the arousal potential of the music score. It is argued...

  3. Micronucleus assay for human peripheral blood lymphocytes as a biomarker of individual sensitivity to assessing radiation health risk in different population

    International Nuclear Information System (INIS)

    Kang, C.-M.; Jeon, H.-J.; Lee, Y.-S.; Lee, S.-J.; Jin, Y.-H.; Kim, Y.-H.; Kim, T.-W.; Cho, C.-K.

    2003-01-01

    Full text: Our studies were to evaluate micronucleus (MN) assay for human peripheral blood lymphocytes (HPBL) as a biomarker of individual sensitivity to assessing radiation health risk in different population in Korea. Further studied are carried out to provide evidence for the existence of individual variations in age-dependent responses. For the MN assay, HPBLs were irradiated with doses of 0, 1, 2, 4, 8Gy 60 Co γ-rays. Spontaneous frequencies not only vary greatly between individuals, but also working or living areas because of the groups with different lifestyle living in different ecological situation and the reaction to radiation exposure. It was shown that the increased level of spontaneous cell with MN was observed with increased age. The relationship between radiosensitivity and the increased spontaneous level of MN may be in inverse proportion. Age and gender are the most important demographic variables impact on MN index with MN frequencies in female being greater than those in male by a factor of depending on the age group. For both sexes, MN frequency was significantly and positively correlated with age. The main lifestyle factors influencing the MN index in subjects are significantly and positively correlated with smoking in measuring the spontaneous frequencies of micronuclei. The described results show that the genetic damaged rate like MN index in human populations is correlated significantly with age, sex and lifestyle factors. So far, it is evident that with regard to the application of MN assay all future studies to evaluate radiation health risks in different population have to take into account the influence of age, gender, and lifestyle. The results suggested that the MN assay have a high potential to ensure appropriate quality control and standard documentation protocol which can be used to monitor a large population exposed to radiation epidemiologically. We conclude that the determination of individual radiosensitivity with MN assay is

  4. Differences in micronucleus induction in peripheral blood reticulocytes of mice exposed to N-ethyl-N-nitrosourea at light and dark dosing times.

    Science.gov (United States)

    Itoh, Keiichi; Masumori, Shoji; Nakajima, Madoka; Hayashi, Makoto; Sakakibara, Hiroyuki; Shimoi, Kayoko

    2012-01-01

    Mammals, including human beings, have a circadian clock system to regulate behavioral and physiological processes. In this study, we investigated the effect of dosing time on micronucleus induction in the bone marrow by evaluating the frequencies of micronucleated peripheral reticulocytes (MNRETs) in mice exposed to N-ethyl-N-nitrosourea (ENU) to assess any difference in genotoxic sensitivity to chemicals between light and dark periods (inactive phase for rodents and active phase for rodents). Male C3H/He mice were treated intraperitoneally with ENU (12.5 or 25 mg/kg body weight) at zeitgeber time (ZT) 3 in the light period or ZT15 in the dark period, and then the time courses of the frequencies of the MNRETs were determined. The frequencies of the MNRETs induced by ENU increased time-dependently and peaked at 48 hr after treatment for ZT3 and ZT15, and were obviously higher in the ZT15 treatment group than the ZT3 treatment group. The MNRETs were measured at 48 hr after treatment with ENU (25 mg/kg body weight) at various dosing times (ZT0, 3, 6, 12, 15 and 18). The frequencies of the MNRETs in mice treated at ZT0, 15 and 18 were significantly higher than those in mice treated at ZT3, 6 and 12. These results suggest that genotoxic sensitivity to chemicals in mouse bone marrow is different between light and dark periods maybe due to different biological responses (detoxification, cell cycle, DNA repair, etc.) related to circadian rhythms.

  5. Use of the fluorescent micronucleus assay to detect the genotoxic effects of radiation and arsenic exposure in exfoliated human epithelial cells

    International Nuclear Information System (INIS)

    Moore, L.E.; Warner, M.L.; Smith, A.H.

    1996-01-01

    The exfoliated cell micronucleus (MN) assay using fluorescent in situ hybridization (FISH) with a centromeric probe is a rapid method for determining the mechanism of MN formation in epithelial tissues exposed to carcinogenic agents. Here, we describe the use of this assay to detect the presence or absence of centromeric DNA in MN induced in vivo by radiation therapy and chronic arsenic (As) ingestion. We examined the buccal cells of an individual receiving 6,500 rads of photon radiation to the head and neck. Exfoliated cells were collected before, during, and after treatment. After radiation exposure a 16.6-fold increase in buccal cell MN frequency was seen. All induced MN were centromere negative (MN-) resulting from chromosome breakage. This finding is consistent with the clastogenic action of radiation and confirmed the reliability of the method. Three weeks post-therapy, MN frequencies returned to baseline. The assay was used on 18 people chronically exposed to high levels of inorganic arsenic (In-As) in drinking water (average level, 1,312 μg As/L) and 18 matched controls (average level, 16 μg As/L). The combined increase in MN frequency was 1.8-fold (P = 0.001, Fisher's exact test). Frequencies of micronuclei containing acentric fragments (MN-) and those containing whole chromosomes (MN+) both increased, suggesting that arsenic may have both clastogenic and weak aneuploidogenic properties in vivo. After stratification on sex, the effect was stronger in male than in female bladder cells. In males the MN-frequency increased 2.06-fold (P =0.07) while the frequency of MN+ increased 1.86-fold (P = 0.08). In addition, the frequencies of MN and MN+ were positively associated with urinary arsenic and its metabolites. The association was stronger for micronuclei containing acentric fragments. By using FISH with centromeric probes, the mechanism of chemically induced genotoxicity can not be determined in epithelial tissues. 35 refs., 4 tabs

  6. Nongenotoxic effects and a reduction of the DXR-induced genotoxic effects of Helianthus annuus Linné (sunflower) seeds revealed by micronucleus assays in mouse bone marrow.

    Science.gov (United States)

    Boriollo, Marcelo Fabiano Gomes; Souza, Luiz Silva; Resende, Marielly Reis; Silva, Thaísla Andrielle da; Oliveira, Nelma de Mello Silva; Resck, Maria Cristina Costa; Dias, Carlos Tadeu dos Santos; Fiorini, João Evangelista

    2014-04-02

    This research evaluated the genotoxicity of oil and tincture of H. annuus L. seeds using the micronucleus assay in bone marrow of mice. The interaction between these preparations and the genotoxic effects of doxorubicin (DXR) was also analysed (antigenotoxicity test). Experimental groups were evaluated at 24-48 h post treatment with N-Nitroso-N-ethylurea (positive control - NEU), DXR (chemotherapeutic), NaCl (negative control), a sunflower tincture (THALS) and two sources of sunflower oils (POHALS and FOHALS). Antigenotoxic assays were carried out using the sunflower tincture and oils separately and in combination with NUE or DXR. For THALS, analysis of the MNPCEs showed no significant differences between treatment doses (250-2,000 mg.Kg-1) and NaCl. A significant reduction in MNPCE was observed when THALS (2,000 mg.Kg-1) was administered in combination with DXR (5 mg.Kg-1). For POHALS or FOHALS, analysis of the MNPCEs also showed no significant differences between treatment doses (250-2,000 mg.Kg-1) and NaCl. However, the combination DXR + POHALS (2,000 mg.Kg-1) or DXR + FOHALS (2,000 mg.Kg-1) not contributed to the MNPCEs reduction. This research suggests absence of genotoxicity of THALS, dose-, time- and sex-independent, and its combination with DXR can reduce the genotoxic effects of DXR. POHALS and FOHALS also showed absence of genotoxicity, but their association with DXR showed no antigenotoxic effects.

  7. No evidence of chromosome damage in children and adolescents with differentiated thyroid carcinoma after receiving {sup 131}I radiometabolic therapy, as evaluated by micronucleus assay and microarray analysis

    Energy Technology Data Exchange (ETDEWEB)

    Federico, Giovanni; Fiore, Lisa; Massart, Francesco; Saggese, Giuseppe [Azienda Ospedaliero-Universitaria Pisana, Department of Pediatrics, Unit of Pediatric Endocrinology and Diabetes, Pisa (Italy); Boni, Giuseppe; Lazzeri, Patrizia; Mariani, Giuliano [Azienda Ospedaliero-Universitaria Pisana, Unit of Nuclear Medicine, Pisa (Italy); Fabiani, Barbara; Verola, Carmela; Scarpato, Roberto [University of Pisa, Department of Biology, Unit of Genetics, Mutagenesis and Environmental Epidemiology, Pisa (Italy); Traino, Claudio [Azienda Ospedaliero-Universitaria Pisana, Health Physics Service, Pisa (Italy)

    2008-11-15

    As {sup 131}I therapy, used to achieve ablation of thyroid gland remnant, can cause chromosome damage in cultured peripheral lymphocytes especially, we investigated whether administration of radioiodine may induce early genome damage in peripheral T lymphocytes of adolescents with differentiated thyroid carcinoma (DTC). We studied 11 patients, aged 14.8 {+-} 3.1 years, who assumed {sup 131}I (range: 1.11-4.44 GBq) to ablate thyroid remnant. A blood sample for micronucleus assay and for evaluating expression of some genes involved in the DNA repair or the apoptosis pathways was obtained from each patient 1 h before (T{sub 0}) and 24 (T{sub 1}) and 48 h (T{sub 2}) post-radioiodine administration. Compared to T{sub 0}, we did not find any difference in the number of micronucleated cells at both T{sub 1} and T{sub 2} in any subject. Nine out of 11 patients had altered expression levels in a majority of the DNA repair and apoptosis genes at T{sub 1}, which decreased at T{sub 2}. We demonstrated for the first time that peripheral cells of DTC children and adolescents who received {sup 131}I at a mean dosage of 3.50 {+-} 0.37 GBq did not show chromosome damage within 48 h from the end of radiometabolic therapy. This may be due to a prompt activation of the cell machinery that maintains the integrity of the genome to prevent harmful double-strand breaks from progressing to chromosome mutations, either by repairing the lesions or by eliminating the most seriously damaged cells via apoptosis. (orig.)

  8. The effect of an optimized imaging flow cytometry analysis template on sample throughput in the reduced culture cytokinesis-block micronucleus assay

    International Nuclear Information System (INIS)

    Rodrigues, M.A.; Beaton-Green, L.A.; Wilkins, R.C.; Probst, C.E.

    2016-01-01

    In cases of overexposure to ionizing radiation, the cytokinesis-block micronucleus (CBMN) assay can be performed in order to estimate the dose of radiation to an exposed individual. However, in the event of a large-scale radiation accident with many potentially exposed casualties, the assay must be able to generate accurate dose estimates to within ±0.5 Gy as quickly as possible. The assay has been adapted to, validated and optimized on the ImageStream"X imaging flow cyto-meter. The ease of running this automated version of the CBMN assay allowed investigation into the accuracy of dose estimates after reducing the volume of whole blood cultured to 200 μl and reducing the culture time to 48 h. The data analysis template used to identify binucleated lymphocyte cells (BNCs) and micronuclei (MN) has since been optimized to improve the sensitivity and specificity of BNC and MN detection. This paper presents a re-analysis of existing data using this optimized analysis template to demonstrate that dose estimations from blinded samples can be obtained to the same level of accuracy in a shorter data collection time. Here, we show that dose estimates from blinded samples were obtained to within ±0.5 Gy of the delivered dose when data collection time was reduced by 30 min at standard culture conditions and by 15 min at reduced culture conditions. Reducing data collection time while retaining the same level of accuracy in our imaging flow cytometry-based version of the CBMN assay results in higher throughput and further increases the relevancy of the CBMN assay as a radiation bio-dosimeter. (authors)

  9. In vitro genotoxicity of neutral red after photo-activation and metabolic activation in the Ames test, the micronucleus test and the comet assay.

    Science.gov (United States)

    Guérard, Melanie; Zeller, Andreas; Singer, Thomas; Gocke, Elmar

    2012-07-04

    Neutral red (Nr) is relatively non-toxic and is widely used as indicator dye in many biological test systems. It absorbs visible light and is known to act as a photosensitizer, involving the generation of reactive oxygen species (type-I reaction) and singlet oxygen (type-II reaction). The mutagenicity of Nr was determined in the Ames test (with Salmonella typhimurium strains TA1535, TA97, TA98, TA98NR, TA100, and TA102) with and without metabolic activation, and with and without photo-activation on agar plates. Similarly to the situation following metabolic activation, photo-mutagenicity of Nr was seen with all Salmonella strains tested, albeit with different effects between these strains. To our knowledge, Nr is the only photo-mutagen showing such a broad action. Since the effects are also observed in strains not known to be responsive to ROS, this indicates that ROS production is not the sole mode of action that leads to photo-genotoxicity. The reactive species produced by irradiation are short-lived as pre-irradiation of an Nr solution did not produce mutagenic effects when added to the bacteria. In addition, mutagenicity in TA98 following irradiation was stronger than in the nitroreductase-deficient strain TA98NR, indicating that nitro derivatives that are transformed by bacterial nitroreductase to hydroxylamines appear to play a role in the photo-mutagenicity of Nr. Photo-genotoxicity of Nr was further investigated in the comet assay and micronucleus test in L5178Y cells. Concentration-dependent increases in primary DNA damage and in the frequency of micronuclei were observed after irradiation. Copyright © 2012 Elsevier B.V. All rights reserved.

  10. Skin scoring in systemic sclerosis

    DEFF Research Database (Denmark)

    Zachariae, Hugh; Bjerring, Peter; Halkier-Sørensen, Lars

    1994-01-01

    Forty-one patients with systemic sclerosis were investigated with a new and simple skin score method measuring the degree of thickening and pliability in seven regions together with area involvement in each region. The highest values were, as expected, found in diffuse cutaneous systemic sclerosis...... (type III SS) and the lowest in limited cutaneous systemic sclerosis (type I SS) with no lesions extending above wrists and ancles. A positive correlation was found to the aminoterminal propeptide of type III procollagen, a serological marker for synthesis of type III collagen. The skin score...

  11. 5-Fluorouracil, colchicine, benzo[a]pyrene and cytosine arabinoside tested in the in vitro mammalian cell micronucleus test (MNvit) in Chinese hamster V79 cells at Covance Laboratories, Harrogate, UK in support of OECD draft Test Guideline 487.

    Science.gov (United States)

    Whitwell, James; Fowler, Paul; Allars, Sarah; Jenner, Karen; Lloyd, Melvyn; Wood, Debbie; Smith, Katie; Young, Jamie; Jeffrey, Laura; Kirkland, David

    2010-10-29

    The reference genotoxic agents 5-fluorouracil (a nucleoside analogue, characterised by a steep dose response profile), colchicine (an aneugen that inhibits tubulin polymerisation), benzo[a]pyrene (a polycyclic aromatic hydrocarbon requiring metabolic activation) and cytosine arabinoside (a nucleoside analogue that inhibits the gap-filling step of excision repair) were tested in the in vitro micronucleus assay using the Chinese hamster V79 cell line at Covance Laboratories, Harrogate, UK. All chemicals were treated in the absence and presence of cytokinesis block (via addition of cytochalasin B) with this work forming part of a collaborative evaluation of the toxicity measures recommended in the draft OECD Test Guideline 487 on the In Vitro Mammalian Cell Micronucleus Test (MNvit). The toxicity measures used, detecting a possible combination of both cytostasis and cell death (though not cell death directly), were relative population doubling, relative increase in cell counts and relative cell counts for treatments in the absence of cytokinesis block, and replication index in the presence of cytokinesis block. All of the chemicals tested either gave marked increases in the percentage of micronucleated cells with and without cytokinesis block, or did not induce micronuclei at concentrations giving approximately 50-60% toxicity (cytostasis and cell death) or less by all of the toxicity measures used. The outcome from this series of tests supports the use of relative increase in cell counts and relative population doubling, as well as relative cell counts, as appropriate measures of cytotoxicity for the non-cytokinesis blocked in vitro micronucleus assay. Copyright © 2010 Elsevier B.V. All rights reserved.

  12. The persistence of depression score

    NARCIS (Netherlands)

    Spijker, J.; de Graaf, R.; Ormel, J.; Nolen, W. A.; Grobbee, D. E.; Burger, H.

    2006-01-01

    Objective: To construct a score that allows prediction of major depressive episode (MDE) persistence in individuals with MDE using determinants of persistence identified in previous research. Method: Data were derived from 250 subjects from the general population with new MDE according to DSM-III-R.

  13. Score distributions in information retrieval

    NARCIS (Netherlands)

    Arampatzis, A.; Robertson, S.; Kamps, J.

    2009-01-01

    We review the history of modeling score distributions, focusing on the mixture of normal-exponential by investigating the theoretical as well as the empirical evidence supporting its use. We discuss previously suggested conditions which valid binary mixture models should satisfy, such as the

  14. Developing Scoring Algorithms (Earlier Methods)

    Science.gov (United States)

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

  15. 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.

  16. 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)

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

    Science.gov (United States)

    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.

  18. 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

  19. 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)

  20. 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.

    Science.gov (United States)

    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.

  1. Combining Teacher Assessment Scores with External Examination ...

    African Journals Online (AJOL)

    Combining Teacher Assessment Scores with External Examination Scores for Certification: Comparative Study of Four Statistical Models. ... University entrance examination scores in mathematics were obtained for a subsample of 115 ...

  2. Scoring System Improvements to Three Leadership Predictors

    National Research Council Canada - National Science Library

    Dela

    1997-01-01

    .... The modified scoring systems were evaluated by rescoring responses randomly selected from the sample which had been scored according to the scoring systems originally developed for the leadership research...

  3. Interpreting force concept inventory scores: Normalized gain and SAT scores

    Directory of Open Access Journals (Sweden)

    Jeffrey J. Steinert

    2007-05-01

    Full Text Available Preinstruction SAT scores and normalized gains (G on the force concept inventory (FCI were examined for individual students in interactive engagement (IE courses in introductory mechanics at one high school (N=335 and one university (N=292 , and strong, positive correlations were found for both populations ( r=0.57 and r=0.46 , respectively. These correlations are likely due to the importance of cognitive skills and abstract reasoning in learning physics. The larger correlation coefficient for the high school population may be a result of the much shorter time interval between taking the SAT and studying mechanics, because the SAT may provide a more current measure of abilities when high school students begin the study of mechanics than it does for college students, who begin mechanics years after the test is taken. In prior research a strong correlation between FCI G and scores on Lawson’s Classroom Test of Scientific Reasoning for students from the same two schools was observed. Our results suggest that, when interpreting class average normalized FCI gains and comparing different classes, it is important to take into account the variation of students’ cognitive skills, as measured either by the SAT or by Lawson’s test. While Lawson’s test is not commonly given to students in most introductory mechanics courses, SAT scores provide a readily available alternative means of taking account of students’ reasoning abilities. Knowing the students’ cognitive level before instruction also allows one to alter instruction or to use an intervention designed to improve students’ cognitive level.

  4. Interpreting force concept inventory scores: Normalized gain and SAT scores

    Directory of Open Access Journals (Sweden)

    Vincent P. Coletta

    2007-05-01

    Full Text Available Preinstruction SAT scores and normalized gains (G on the force concept inventory (FCI were examined for individual students in interactive engagement (IE courses in introductory mechanics at one high school (N=335 and one university (N=292, and strong, positive correlations were found for both populations (r=0.57 and r=0.46, respectively. These correlations are likely due to the importance of cognitive skills and abstract reasoning in learning physics. The larger correlation coefficient for the high school population may be a result of the much shorter time interval between taking the SAT and studying mechanics, because the SAT may provide a more current measure of abilities when high school students begin the study of mechanics than it does for college students, who begin mechanics years after the test is taken. In prior research a strong correlation between FCI G and scores on Lawson’s Classroom Test of Scientific Reasoning for students from the same two schools was observed. Our results suggest that, when interpreting class average normalized FCI gains and comparing different classes, it is important to take into account the variation of students’ cognitive skills, as measured either by the SAT or by Lawson’s test. While Lawson’s test is not commonly given to students in most introductory mechanics courses, SAT scores provide a readily available alternative means of taking account of students’ reasoning abilities. Knowing the students’ cognitive level before instruction also allows one to alter instruction or to use an intervention designed to improve students’ cognitive level.

  5. Blind Grid Scoring Record No. 290

    National Research Council Canada - National Science Library

    Overbay, Larry; Robitaille, George

    2005-01-01

    ...) utilizing the APG Standardized UXO Technology Demonstration Site Blind Grid. Scoring Records have been coordinated by Larry Overbay and the Standardized UXO Technology Demonstration Site Scoring Committee...

  6. Blind Grid Scoring Record No. 293

    National Research Council Canada - National Science Library

    Overbay, Larry; Robitaille, George; Archiable, Robert; Fling, Rick; McClung, Christina

    2005-01-01

    ...) utilizing the YPG Standardized UXO Technology Demonstration Site Blind Grid. Scoring Records have been coordinated by Larry Overbay and the Standardized UXO Technology Demonstration Site Scoring Committee...

  7. Open Field Scoring Record No. 298

    National Research Council Canada - National Science Library

    Overbay, Jr., Larry; Robitaille, George; Fling, Rick; McClung, Christina

    2005-01-01

    ...) utilizing the APG Standardized UXO Technology Demonstration Site Open Field. Scoring Records have been coordinated by Larry Overbay and the Standardized UXO Technology Demonstration Site Scoring Committee...

  8. Open Field Scoring Record No. 299

    National Research Council Canada - National Science Library

    Overbay, Larry; Robitaille, George

    2005-01-01

    ...) utilizing the YPG Standardized UXO Technology Demonstration Site Open Field. Scoring Records have been coordinated by Larry Overbay and the standardized UXO Technology Demonstration Site Scoring Committee...

  9. Interval Coded Scoring: a toolbox for interpretable scoring systems

    Directory of Open Access Journals (Sweden)

    Lieven Billiet

    2018-04-01

    Full Text Available Over the last decades, clinical decision support systems have been gaining importance. They help clinicians to make effective use of the overload of available information to obtain correct diagnoses and appropriate treatments. However, their power often comes at the cost of a black box model which cannot be interpreted easily. This interpretability is of paramount importance in a medical setting with regard to trust and (legal responsibility. In contrast, existing medical scoring systems are easy to understand and use, but they are often a simplified rule-of-thumb summary of previous medical experience rather than a well-founded system based on available data. Interval Coded Scoring (ICS connects these two approaches, exploiting the power of sparse optimization to derive scoring systems from training data. The presented toolbox interface makes this theory easily applicable to both small and large datasets. It contains two possible problem formulations based on linear programming or elastic net. Both allow to construct a model for a binary classification problem and establish risk profiles that can be used for future diagnosis. All of this requires only a few lines of code. ICS differs from standard machine learning through its model consisting of interpretable main effects and interactions. Furthermore, insertion of expert knowledge is possible because the training can be semi-automatic. This allows end users to make a trade-off between complexity and performance based on cross-validation results and expert knowledge. Additionally, the toolbox offers an accessible way to assess classification performance via accuracy and the ROC curve, whereas the calibration of the risk profile can be evaluated via a calibration curve. Finally, the colour-coded model visualization has particular appeal if one wants to apply ICS manually on new observations, as well as for validation by experts in the specific application domains. The validity and applicability

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

    Energy Technology Data Exchange (ETDEWEB)

    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

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

    Science.gov (United States)

    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

  12. [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%.

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

    Directory of Open Access Journals (Sweden)

    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

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

    Science.gov (United States)

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

    2018-01-01

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

  15. 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.

  16. Exploring a Source of Uneven Score Equity across the Test Score Range

    Science.gov (United States)

    Huggins-Manley, Anne Corinne; Qiu, Yuxi; Penfield, Randall D.

    2018-01-01

    Score equity assessment (SEA) refers to an examination of population invariance of equating across two or more subpopulations of test examinees. Previous SEA studies have shown that score equity may be present for examinees scoring at particular test score ranges but absent for examinees scoring at other score ranges. No studies to date have…

  17. Response of thyroid follicular cells to gamma irradiation compared to proton irradiation. I. Initial characterization of DNA damage, micronucleus formation, apoptosis, cell survival, and cell cycle phase redistribution

    Science.gov (United States)

    Green, L. M.; Murray, D. K.; Bant, A. M.; Kazarians, G.; Moyers, M. F.; Nelson, G. A.; Tran, D. T.

    2001-01-01

    The RBE of protons has been assumed to be equivalent to that of photons. The objective of this study was to determine whether radiation-induced DNA and chromosome damage, apoptosis, cell killing and cell cycling in organized epithelial cells was influenced by radiation quality. Thyroid-stimulating hormone-dependent Fischer rat thyroid cells, established as follicles, were exposed to gamma rays or proton beams delivered acutely over a range of physical doses. Gamma-irradiated cells were able to repair DNA damage relatively rapidly so that by 1 h postirradiation they had approximately 20% fewer exposed 3' ends than their counterparts that had been irradiated with proton beams. The persistence of free ends of DNA in the samples irradiated with the proton beam implies that either more initial breaks or a quantitatively different type of damage had occurred. These results were further supported by an increased frequency of chromosomal damage as measured by the presence of micronuclei. Proton-beam irradiation induced micronuclei at a rate of 2.4% per gray, which at 12 Gy translated to 40% more micronuclei than in comparable gamma-irradiated cultures. The higher rate of micronucleus formation and the presence of larger micronuclei in proton-irradiated cells was further evidence that a qualitatively more severe class of damage had been induced than was induced by gamma rays. Differences in the type of damage produced were detected in the apoptosis assay, wherein a significant lag in the induction of apoptosis occurred after gamma irradiation that did not occur with protons. The more immediate expression of apoptotic cells in the cultures irradiated with the proton beam suggests that the damage inflicted was more severe. Alternatively, the cell cycle checkpoint mechanisms required for recovery from such damage might not have been invoked. Differences based on radiation quality were also evident in the alpha components of cell survival curves (0.05 Gy(-1) for gamma rays, 0

  18. Linkage between company scores and stock returns

    Directory of Open Access Journals (Sweden)

    Saban Celik

    2017-12-01

    Full Text Available Previous studies on company scores conducted at firm-level, generally concluded that there exists a positive relation between company scores and stock returns. Motivated by these studies, this study examines the relationship between company scores (Corporate Governance Score, Economic Score, Environmental Score, and Social Score and stock returns, both at portfolio-level analysis and firm-level cross-sectional regressions. In portfolio-level analysis, stocks are sorted based on each company scores and quintile portfolio are formed with different levels of company scores. Then, existence and significance of raw returns and risk-adjusted returns difference between portfolios with the extreme company scores (portfolio 10 and portfolio 1 is tested. In addition, firm-level cross-sectional regression is performed to examine the significance of company scores effects with control variables. While portfolio-level analysis results indicate that there is no significant relation between company scores and stock returns; firm-level analysis indicates that economic, environmental, and social scores have effect on stock returns, however, significance and direction of these effects change, depending on the included control variables in the cross-sectional regression.

  19. 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

  20. Cardiovascular risk scores for coronary atherosclerosis.

    Science.gov (United States)

    Yalcin, Murat; Kardesoglu, Ejder; Aparci, Mustafa; Isilak, Zafer; Uz, Omer; Yiginer, Omer; Ozmen, Namik; Cingozbay, Bekir Yilmaz; Uzun, Mehmet; Cebeci, Bekir Sitki

    2012-10-01

    The objective of this study was to compare frequently used cardiovascular risk scores in predicting the presence of coronary artery disease (CAD) and 3-vessel disease. In 350 consecutive patients (218 men and 132 women) who underwent coronary angiography, the cardiovascular risk level was determined using the Framingham Risk Score (FRS), the Modified Framingham Risk Score (MFRS), the Prospective Cardiovascular Münster (PROCAM) score, and the Systematic Coronary Risk Evaluation (SCORE). The area under the curve for receiver operating characteristic curves showed that FRS had more predictive value than the other scores for CAD (area under curve, 0.76, P MFRS, PROCAM, and SCORE) may predict the presence and severity of coronary atherosclerosis.The FRS had better predictive value than the other scores.

  1. Lack of micronucleus induction activity of ethyl tertiary-butyl ether in the bone marrow of F344 rats by sub-chronic drinking-water treatment, inhalation exposure, or acute intraperitoneal injection.

    Science.gov (United States)

    Noguchi, Tadashi; Kamigaito, Tomoyuki; Katagiri, Taku; Kondou, Hitomi; Yamazaki, Kazunori; Aiso, Shigetoshi; Nishizawa, Tomoshi; Nagano, Kasuke; Fukushima, Shoji

    2013-01-01

    Ethyl tertiary-butyl ether (ETBE) is an oxygenated gasoline additive synthesized from ethanol and isobutene that is used to reduce CO2 emissions. To support the Kyoto Protocol, the production of ETBE has undergone a marked increase. Previous reports have indicated that exposure to ETBE or methyl tertiary-butyl ether resulted in liver and kidney tumors in rats and/or mice. These reports raise concern about the effects of human exposure being brought about by the increased use of ETBE. The present study was conducted to evaluate the genotoxicity of ETBE using micronucleus induction of polychromatic erythrocytes in the bone marrow of male and female rats treated with ETBE in the drinking-water at concentrations of 0, 1,600, 4,000 or 10,000 ppm or exposed to ETBE vapor at 0, 500, 1,500 or 5,000 ppm for 13 weeks. There were no significant increases in micronucleus induction in either the drinking water-administered or inhalation-administered groups at any concentration of ETBE; although, in both groups red blood cells and hemoglobin concentration were slightly reduced in the peripheral blood in rats administered the highest concentration of ETBE. In addition, two consecutive daily intraperitoneal injections of ETBE at doses of 0, 250, 500 or 1,000 mg/kg did not increase the frequency of micronucleated bone marrow cells in either sex; all rats receiving intraperitoneal injections of ETBE at a dose of 2,000 mg/kg died after treatment day 1. These data suggest that ETBE is not genotoxic in vivo.

  2. Interobserver variability of the neurological optimality score

    NARCIS (Netherlands)

    Monincx, W. M.; Smolders-de Haas, H.; Bonsel, G. J.; Zondervan, H. A.

    1999-01-01

    To assess the interobserver reliability of the neurological optimality score. The neurological optimality score of 21 full term healthy, neurologically normal newborn infants was determined by two well trained observers. The interclass correlation coefficient was 0.31. Kappa for optimality (score of

  3. Semiparametric score level fusion: Gaussian copula approach

    NARCIS (Netherlands)

    Susyanyo, N.; Klaassen, C.A.J.; Veldhuis, Raymond N.J.; Spreeuwers, Lieuwe Jan

    2015-01-01

    Score level fusion is an appealing method for combining multi-algorithms, multi- representations, and multi-modality biometrics due to its simplicity. Often, scores are assumed to be independent, but even for dependent scores, accord- ing to the Neyman-Pearson lemma, the likelihood ratio is the

  4. An Objective Fluctuation Score for Parkinson's Disease

    Science.gov (United States)

    Horne, Malcolm K.; McGregor, Sarah; Bergquist, Filip

    2015-01-01

    Introduction Establishing the presence and severity of fluctuations is important in managing Parkinson’s Disease yet there is no reliable, objective means of doing this. In this study we have evaluated a Fluctuation Score derived from variations in dyskinesia and bradykinesia scores produced by an accelerometry based system. Methods The Fluctuation Score was produced by summing the interquartile range of bradykinesia scores and dyskinesia scores produced every 2 minutes between 0900-1800 for at least 6 days by the accelerometry based system and expressing it as an algorithm. Results This Score could distinguish between fluctuating and non-fluctuating patients with high sensitivity and selectivity and was significant lower following activation of deep brain stimulators. The scores following deep brain stimulation lay in a band just above the score separating fluctuators from non-fluctuators, suggesting a range representing adequate motor control. When compared with control subjects the score of newly diagnosed patients show a loss of fluctuation with onset of PD. The score was calculated in subjects whose duration of disease was known and this showed that newly diagnosed patients soon develop higher scores which either fall under or within the range representing adequate motor control or instead go on to develop more severe fluctuations. Conclusion The Fluctuation Score described here promises to be a useful tool for identifying patients whose fluctuations are progressing and may require therapeutic changes. It also shows promise as a useful research tool. Further studies are required to more accurately identify therapeutic targets and ranges. PMID:25928634

  5. Breaking of scored tablets : a review

    NARCIS (Netherlands)

    van Santen, E; Barends, D M; Frijlink, H W

    The literature was reviewed regarding advantages, problems and performance indicators of score lines. Scored tablets provide dose flexibility, ease of swallowing and may reduce the costs of medication. However, many patients are confronted with scored tablets that are broken unequally and with

  6. Validation of Automated Scoring of Science Assessments

    Science.gov (United States)

    Liu, Ou Lydia; Rios, Joseph A.; Heilman, Michael; Gerard, Libby; Linn, Marcia C.

    2016-01-01

    Constructed response items can both measure the coherence of student ideas and serve as reflective experiences to strengthen instruction. We report on new automated scoring technologies that can reduce the cost and complexity of scoring constructed-response items. This study explored the accuracy of c-rater-ML, an automated scoring engine…

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

    Science.gov (United States)

    Dasgupta, Alakananda; Mahapatra, Manoranjan; Saxena, Renu

    2013-04-01

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

  8. Oswestry Disability Index scoring made easy.

    Science.gov (United States)

    Mehra, A; Baker, D; Disney, S; Pynsent, P B

    2008-09-01

    Low back pain effects up to 80% of the population at some time during their active life. Questionnaires are available to help measure pain and disability. The Oswestry Disability Index (ODI) is the most commonly used outcome measure for low back pain. The aim of this study was to see if training in completing the ODI forms improved the scoring accuracy. The last 100 ODI forms completed in a hospital's spinal clinic were reviewed retrospectively and errors in the scoring were identified. Staff members involved in scoring the questionnaire were made aware of the errors and the correct method of scoring explained. A chart was created with all possible scores to aid the staff with scoring. A prospective audit on 50 questionnaires was subsequently performed. The retrospective study showed that 33 of the 100 forms had been incorrectly scored. All questionnaires where one or more sections were not completed by the patient were incorrectly scored. A scoring chart was developed and staff training was implemented. This reduced the error rate to 14% in the prospective audit. Clinicians applying outcome measures should read the appropriate literature to ensure they understand the scoring system. Staff must then be given adequate training in the application of the questionnaires.

  9. Combination of scoring schemes for protein docking

    Directory of Open Access Journals (Sweden)

    Schomburg Dietmar

    2007-08-01

    Full Text Available Abstract Background Docking algorithms are developed to predict in which orientation two proteins are likely to bind under natural conditions. The currently used methods usually consist of a sampling step followed by a scoring step. We developed a weighted geometric correlation based on optimised atom specific weighting factors and combined them with our previously published amino acid specific scoring and with a comprehensive SVM-based scoring function. Results The scoring with the atom specific weighting factors yields better results than the amino acid specific scoring. In combination with SVM-based scoring functions the percentage of complexes for which a near native structure can be predicted within the top 100 ranks increased from 14% with the geometric scoring to 54% with the combination of all scoring functions. Especially for the enzyme-inhibitor complexes the results of the ranking are excellent. For half of these complexes a near-native structure can be predicted within the first 10 proposed structures and for more than 86% of all enzyme-inhibitor complexes within the first 50 predicted structures. Conclusion We were able to develop a combination of different scoring schemes which considers a series of previously described and some new scoring criteria yielding a remarkable improvement of prediction quality.

  10. Forecasting the value of credit scoring

    Science.gov (United States)

    Saad, Shakila; Ahmad, Noryati; Jaffar, Maheran Mohd

    2017-08-01

    Nowadays, credit scoring system plays an important role in banking sector. This process is important in assessing the creditworthiness of customers requesting credit from banks or other financial institutions. Usually, the credit scoring is used when customers send the application for credit facilities. Based on the score from credit scoring, bank will be able to segregate the "good" clients from "bad" clients. However, in most cases the score is useful at that specific time only and cannot be used to forecast the credit worthiness of the same applicant after that. Hence, bank will not know if "good" clients will always be good all the time or "bad" clients may become "good" clients after certain time. To fill up the gap, this study proposes an equation to forecast the credit scoring of the potential borrowers at a certain time by using the historical score related to the assumption. The Mean Absolute Percentage Error (MAPE) is used to measure the accuracy of the forecast scoring. Result shows the forecast scoring is highly accurate as compared to actual credit scoring.

  11. Development of the siriraj clinical asthma score.

    Science.gov (United States)

    Vichyanond, Pakit; Veskitkul, Jittima; Rienmanee, Nuanphong; Pacharn, Punchama; Jirapongsananuruk, Orathai; Visitsunthorn, Nualanong

    2013-09-01

    Acute asthmatic attack in children commonly occurs despite the introduction of effective controllers such as inhaled corticosteroids and leukotriene modifiers. Treatment of acute asthmatic attack requires proper evaluation of attack severity and appropriate selection of medical therapy. In children, measurement of lung function is difficult during acute attack and thus clinical asthma scoring may aid physician in making further decision regarding treatment and admission. We enrolled 70 children with acute asthmatic attack with age range from 1 to 12 years (mean ± SD = 51.5 ± 31.8 months) into the study. Twelve selected asthma severity items were assessed by 2 independent observers prior to administration of salbutamol nebulization (up to 3 doses at 20 minutes interval). Decision for further therapy and admission was made by emergency department physician. Three different scoring systems were constructed from items with best validity. Sensitivity, specificity and accuracy of these scores were assessed. Inter-rater reliability was assessed for each score. Review of previous scoring systems was also conducted and reported. Three severity items had poor validity, i.e., cyanosis, depressed cerebral function, and I:E ratio (p > 0.05). Three items had poor inter-rater reliability, i.e., breath sound quality, air entry, and I:E ratio. These items were omitted and three new clinical scores were constructed from the remaining items. Clinical scoring system comprised retractions, dyspnea, O2 saturation, respiratory rate and wheezing (rangeof score 0-10) gave the best accuracy and inter-rater variability and were chosen for clinical use-Siriraj Clinical Asthma Score (SCAS). A Clinical Asthma Score that is simple, relatively easy to administer and with good validity and variability is essential for treatment of acute asthma in children. Several good candidate scores have been introduced in the past. We described the development of the Siriraj Clinical Asthma Score (SCAS) in

  12. A diagnostic scoring system for myxedema coma.

    Science.gov (United States)

    Popoveniuc, Geanina; Chandra, Tanu; Sud, Anchal; Sharma, Meeta; Blackman, Marc R; Burman, Kenneth D; Mete, Mihriye; Desale, Sameer; Wartofsky, Leonard

    2014-08-01

    To develop diagnostic criteria for myxedema coma (MC), a decompensated state of extreme hypothyroidism with a high mortality rate if untreated, in order to facilitate its early recognition and treatment. The frequencies of characteristics associated with MC were assessed retrospectively in patients from our institutions in order to derive a semiquantitative diagnostic point scale that was further applied on selected patients whose data were retrieved from the literature. Logistic regression analysis was used to test the predictive power of the score. Receiver operating characteristic (ROC) curve analysis was performed to test the discriminative power of the score. Of the 21 patients examined, 7 were reclassified as not having MC (non-MC), and they were used as controls. The scoring system included a composite of alterations of thermoregulatory, central nervous, cardiovascular, gastrointestinal, and metabolic systems, and presence or absence of a precipitating event. All 14 of our MC patients had a score of ≥60, whereas 6 of 7 non-MC patients had scores of 25 to 50. A total of 16 of 22 MC patients whose data were retrieved from the literature had a score ≥60, and 6 of 22 of these patients scored between 45 and 55. The odds ratio per each score unit increase as a continuum was 1.09 (95% confidence interval [CI], 1.01 to 1.16; P = .019); a score of 60 identified coma, with an odds ratio of 1.22. The area under the ROC curve was 0.88 (95% CI, 0.65 to 1.00), and the score of 60 had 100% sensitivity and 85.71% specificity. A score ≥60 in the proposed scoring system is potentially diagnostic for MC, whereas scores between 45 and 59 could classify patients at risk for MC.

  13. A Comparison of Two Scoring Methods for an Automated Speech Scoring System

    Science.gov (United States)

    Xi, Xiaoming; Higgins, Derrick; Zechner, Klaus; Williamson, David

    2012-01-01

    This paper compares two alternative scoring methods--multiple regression and classification trees--for an automated speech scoring system used in a practice environment. The two methods were evaluated on two criteria: construct representation and empirical performance in predicting human scores. The empirical performance of the two scoring models…

  14. [The diagnostic scores for deep venous thrombosis].

    Science.gov (United States)

    Junod, A

    2015-08-26

    Seven diagnostic scores for the deep venous thrombosis (DVT) of lower limbs are analyzed and compared. Two features make this exer- cise difficult: the problem of distal DVT and of their proximal extension and the status of patients, whether out- or in-patients. The most popular score is the Wells score (1997), modi- fied in 2003. It includes one subjective ele- ment based on clinical judgment. The Primary Care score 12005), less known, has similar pro- perties, but uses only objective data. The pre- sent trend is to associate clinical scores with the dosage of D-Dimers to rule out with a good sensitivity the probability of TVP. For the upper limb DVT, the Constans score (2008) is available, which can also be coupled with D-Dimers testing (Kleinjan).

  15. Scoring an Abstract Contemporary Silent Film

    OpenAIRE

    Frost, Crystal

    2014-01-01

    I composed an original digital audio film score with full sound design for a contemporary silent film called Apple Tree. The film is highly conceptual and interpretive and required a very involved, intricate score to successfully tell the story. In the process of scoring this film, I learned new ways to convey an array of contrasting emotions through music and sound. After analyzing the film's emotional journey, I determined that six defining emotions were the foundation on which to build an ...

  16. The FAt Spondyloarthritis Spine Score (FASSS)

    DEFF Research Database (Denmark)

    Pedersen, Susanne Juhl; Zhao, Zheng; Lambert, Robert Gw

    2013-01-01

    an important measure of treatment efficacy as well as a surrogate marker for new bone formation. The aim of this study was to develop and validate a new scoring method for fat lesions in the spine, the Fat SpA Spine Score (FASSS), which in contrast to the existing scoring method addresses the localization......Studies have shown that fat lesions follow resolution of inflammation in the spine of patients with axial spondyloarthritis (SpA). Fat lesions at vertebral corners have also been shown to predict development of new syndesmophytes. Therefore, scoring of fat lesions in the spine may constitute both...

  17. Recognition Using Classification and Segmentation Scoring

    National Research Council Canada - National Science Library

    Kimball, Owen; Ostendorf, Mari; Rohlicek, Robin

    1992-01-01

    .... We describe an approach to connected word recognition that allows the use of segmental information through an explicit decomposition of the recognition criterion into classification and segmentation scoring...

  18. Equating error in observed-score equating

    NARCIS (Netherlands)

    van der Linden, Willem J.

    2006-01-01

    Traditionally, error in equating observed scores on two versions of a test is defined as the difference between the transformations that equate the quantiles of their distributions in the sample and population of test takers. But it is argued that if the goal of equating is to adjust the scores of

  19. Correlating continuous assessment scores to junior secondary ...

    African Journals Online (AJOL)

    This study investigated the relationship between continuous assessment scores and junior secondary school certificate examination(JSCE) final scores in Imo State. A sample of four hundred students were purposively selected from thirty eight thousand students who took the 1997 JSCE in Imo State. The data used were ...

  20. Summary of Score Changes (in other Tests).

    Science.gov (United States)

    Cleary, T. Anne; McCandless, Sam A.

    Scholastic Aptitude Test (SAT) scores have declined during the last 14 years. Similar score declines have been observed in many different testing programs, many groups, and tested areas. The declines, while not large in any given year, have been consistent over time, area, and group. The period around 1965 is critical for the interpretation of…

  1. More Issues in Observed-Score Equating

    Science.gov (United States)

    van der Linden, Wim J.

    2013-01-01

    This article is a response to the commentaries on the position paper on observed-score equating by van der Linden (this issue). The response focuses on the more general issues in these commentaries, such as the nature of the observed scores that are equated, the importance of test-theory assumptions in equating, the necessity to use multiple…

  2. Clinical scoring scales in thyroidology: A compendium

    Directory of Open Access Journals (Sweden)

    Sanjay Kalra

    2011-01-01

    Full Text Available This compendium brings together traditional as well as contemporary scoring and grading systems used for the screening and diagnosis of various thyroid diseases, dysfunctions, and complications. The article discusses scores used to help diagnose hypo-and hyperthyroidism, to grade and manage goiter and ophthalmopathy, and to assess the risk of thyroid malignancy.

  3. Semiparametric Copula Models for Biometric Score Level

    NARCIS (Netherlands)

    Caselli, M.

    2016-01-01

    In biometric recognition systems, biometric samples (images of faces, finger- prints, voices, gaits, etc.) of people are compared and classifiers (matchers) indicate the level of similarity between any pair of samples by a score. If two samples of the same person are compared, a genuine score is

  4. Intelligence Score Profiles of Female Juvenile Offenders

    Science.gov (United States)

    Werner, Shelby Spare; Hart, Kathleen J.; Ficke, Susan L.

    2016-01-01

    Previous studies have found that male juvenile offenders typically obtain low scores on measures of intelligence, often with a pattern of higher scores on measures of nonverbal relative to verbal tasks. The research on the intelligence performance of female juvenile offenders is limited. This study explored the Wechsler Intelligence Scale for…

  5. In vitro mutagenicity and genotoxicity study of 1,2-dichloroethylene, 1,1,2-trichloroethane, 1,3-dichloropropane, 1,2,3-trichloropropane and 1,1,3-trichloropropene, using the micronucleus test and the alkaline single cell gel electrophoresis technique (comet assay) in human lymphocytes.

    Science.gov (United States)

    Tafazoli, M; Kirsch-Volders, M

    1996-12-20

    The main objective of this study was to compare the cytotoxic genotoxic and mutagenic activity of a number of chlorinated aliphatic hydrocarbons, which are widely used as chemical intermediates, solvents, degreasing agents etc. in industry, and to establish the structure-toxicity relationship of the chemicals by using the most adequate determinants in estimating their toxicity. The mutagenicity and cytotoxicity of some of the candidate chemicals, namely 1,2-dichloroethylene, 1,1,2-trichloroethane, 1,3-dichloropropane, 1,2,3-trichloropropane and 1,1,3-trichloropropene were evaluated in an in vitro micronucleus assay. The cytokinesis-block methodology was applied on human lymphocytes in the presence or absence of an external metabolic activation system (S9-mix). In the micronucleus assay, all test substances, except 1,2,3-trichloropropane with and without S9-mix and 1,1,2-trichloroethane without S9-mix in the repeated experiment, exhibited a low but statistically significant mutagenic activity, compared to the concurrent control. However, none of the five chemicals was able to induce a clear and reproducible linear dose-dependent increase in micronucleus frequencies in this assay. Generally, mutagenic activity of the chemicals was found in the absence of severe cytotoxicity and/or cell cycle delay. The DNA breakage capacity and the cytotoxicity of these chemicals were also assessed in the alkaline single cell gel (SCG) electrophoresis test (comet assay) with and without S9-mix in isolated human lymphocytes. All chemical compounds induced DNA breakage, in the presence or absence of the metabolic activation system, at the doses tested. The data showed that the DNA reactivity of the chemicals increased with increasing degree of halogenation. The results of the present work suggested that the comet assay might be a more suitable and sensitive screening method than the micronucleus test for this particular class of compound. However, both assays do detect different

  6. Study on cell survival, induction of apoptosis and micronucleus formation in SCL-II and RTiV3 cells after exposure to the Auger electron emitter Tc-99m

    International Nuclear Information System (INIS)

    Kadenbach, K.; Kriehuber, R.; Weiss, D.G.

    2003-01-01

    Full text: Cell survival, induction of apoptosis and micronucleus (MN) formation have been investigated in the human squamous cell carcinoma cell line SCL-II and in the rat tracheal cell line RTiV3 after exposure to the Auger electron emitter Tc-99m. Cells were either acutely gamma(Co-60)-irradiated (0.78 Gy/min) or exposed to Tc-99m-Pertechnetate (25-300 MBq/20ml) for 24 h under cell culture conditions and assayed for cell survival (Colony-forming assay), micronucleus formation (Cytochalasin B assay) and the frequency of apoptotic cells (Fluorescence microscopy). Analytical dosimetrical models have been applied to derive the absorbed dose corresponding to the accumulated decays of Tc-99m. Absorbed doses up to 1.3 Gy could be achieved after Tc-99m exposure leading to no significant cell killing in this dose range except at one dose point (0.25 Gy) in SCL-II cells. MN formation was consistently lower when compared to Co-60 irradiated cells and showed a linear dose-response. The apoptotic response in SCL-II cells after Tc-99m exposure was described best by a 3rd order polynomial and increased apoptosis induction could be observed at much lower doses (0.25 Gy) in comparison to the reference radiation (0.8 Gy). The relative biological effectiveness (RBE) has been determined for MN formation and apoptosis induction and was found to be in the range of 0.1- 1.3 for both investigated biological endpoints, depending on which mathematical model for describing the dose-effect curve was used. Up-take experiments revealed an activity concentration ratio cells vs. medium of 1.2 after 16 h up to 24 h of exposure. No increased biological effectiveness of Tc-99m applied as Sodium-Pertechnetate could be observed in the investigated cell lines in comparison to gamma-irradiation. Induction of apoptosis is slightly increased after Tc-99m exposure in SCL-II cells and it has to be further evaluated, if this is due to the emitted Auger-component. A passive up-take mechanism of Tc-99m is

  7. [The use of scores in general medicine].

    Science.gov (United States)

    Huber, Ursula; Rösli, Andreas; Ballmer, Peter E; Rippin, Sarah Jane

    2013-10-01

    Scores are tools to combine complex information into a numerical value. In General Medicine, there are scores to assist in making diagnoses and prognoses, scores to assist therapeutic decision making and to evaluate therapeutic results and scores to help physicians when informing and advising patients. We review six of the scoring systems that have the greatest utility for the General Physician in hospital-based care and in General Practice. The Nutritional Risk Screening (NRS 2002) tool is designed to identify hospital patients in danger of malnutrition. The aim is to improve the nutritional status of these patients. The CURB-65 score predicts 30-day mortality in patients with community acquired pneumonia. Patients with a low score can be considered for home treatment, patients with an elevated score require hospitalisation and those with a high score should be treated as having severe pneumonia; treatment in the intensive care unit should be considered. The IAS-AGLA score of the Working Group on Lipids and Atherosclerosis of the Swiss Society of Cardiology calculates the 10-year risk of a myocardial infarction for people living in Switzerland. The working group makes recommendations for preventative treatment according to the calculated risk status. The Body Mass Index, which is calculated by dividing the body weight in kilograms by the height in meters squared and then divided into weight categories, is used to classify people as underweight, of normal weight, overweight or obese. The prognostic value of this classification is discussed. The Mini-Mental State Examination allows the physician to assess important cognitive functions in a simple and standardised form. The Glasgow Coma Scale is used to classify the level of consciousness in patients with head injury. It can be used for triage and correlates with prognosis.

  8. 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.

  9. THE EFFICIENCY OF TENNIS DOUBLES SCORING SYSTEMS

    Directory of Open Access Journals (Sweden)

    Geoff Pollard

    2010-09-01

    Full Text Available In this paper a family of scoring systems for tennis doubles for testing the hypothesis that pair A is better than pair B versus the alternative hypothesis that pair B is better than A, is established. This family or benchmark of scoring systems can be used as a benchmark against which the efficiency of any doubles scoring system can be assessed. Thus, the formula for the efficiency of any doubles scoring system is derived. As in tennis singles, one scoring system based on the play-the-loser structure is shown to be more efficient than the benchmark systems. An expression for the relative efficiency of two doubles scoring systems is derived. Thus, the relative efficiency of the various scoring systems presently used in doubles can be assessed. The methods of this paper can be extended to a match between two teams of 2, 4, 8, …doubles pairs, so that it is possible to establish a measure for the relative efficiency of the various systems used for tennis contests between teams of players.

  10. A comparison between modified Alvarado score and RIPASA score in the diagnosis of acute appendicitis.

    Science.gov (United States)

    Singla, Anand; Singla, Satpaul; Singh, Mohinder; Singla, Deeksha

    2016-12-01

    Acute appendicitis is a common but elusive surgical condition and remains a diagnostic dilemma. It has many clinical mimickers and diagnosis is primarily made on clinical grounds, leading to the evolution of clinical scoring systems for pin pointing the right diagnosis. The modified Alvarado and RIPASA scoring systems are two important scoring systems, for diagnosis of acute appendicitis. We prospectively compared the two scoring systems for diagnosing acute appendicitis in 50 patients presenting with right iliac fossa pain. The RIPASA score correctly classified 88 % of patients with histologically confirmed acute appendicitis compared with 48.0 % with modified Alvarado score, indicating that RIPASA score is more superior to Modified Alvarado score in our clinical settings.

  11. Facilitating the Interpretation of English Language Proficiency Scores: Combining Scale Anchoring and Test Score Mapping Methodologies

    Science.gov (United States)

    Powers, Donald; Schedl, Mary; Papageorgiou, Spiros

    2017-01-01

    The aim of this study was to develop, for the benefit of both test takers and test score users, enhanced "TOEFL ITP"® test score reports that go beyond the simple numerical scores that are currently reported. To do so, we applied traditional scale anchoring (proficiency scaling) to item difficulty data in order to develop performance…

  12. In situ biomonitoring of the genotoxic effects of mixed industrial emissions using the Tradescantia micronucleus and pollen abortion tests with wild life plants: Demonstration of the efficacy of emission controls in an eastern European city

    Energy Technology Data Exchange (ETDEWEB)

    Misik, Miroslav [Department of Botany, Comenius University in Bratislava, Faculty of Natural Sciences, Revova 39, SK 811 02 Bratislava 1 (Slovakia); Micieta, Karol [Department of Botany, Comenius University in Bratislava, Faculty of Natural Sciences, Revova 39, SK 811 02 Bratislava 1 (Slovakia); Solenska, Martina [Department of Botany, Comenius University in Bratislava, Faculty of Natural Sciences, Revova 39, SK 811 02 Bratislava 1 (Slovakia); Misikova, Katarina [Department of Botany, Comenius University in Bratislava, Faculty of Natural Sciences, Revova 39, SK 811 02 Bratislava 1 (Slovakia); Pisarcikova, Helena [Department of Botany, Comenius University in Bratislava, Faculty of Natural Sciences, Revova 39, SK 811 02 Bratislava 1 (Slovakia); Knasmueller, Siegfried [Institute of Cancer Research, Department of Inner Medicine I, Medical University of Vienna, Borschkegasse 8a, A-1090 Vienna (Austria)]. E-mail: siegfried.knasmueller@meduniwien.ac.at

    2007-01-15

    Aim of the study was to monitor changes of genotoxic activity of urban air caused by an incinerator and a petrochemical plant in Tradescantia micronucleus (Trad-MCN) and pollen fertility assays with wild plants (Chelidonium majus, Clematis vitalba, Cichorium intybus, Linaria vulgaris, Robinia pseudoacacia). While in the first sampling period (1997-2000) significantly (on average 80%) more MN were found at the polluted site in comparison to controls from a rural area, no significant effects were observed during a later period (between 2003 and 2005). A similar pattern was observed in the pollen abortion assays in which the most pronounced effects were found in chicory and false acacia. The differences of the results obtained in the two periods can be explained by a substantial reduction of air pollution by use of new technologies. In particular the decrease of SO{sub 2} emissions may account for the effects seen in the present study. - Air pollution caused by industrial emissions induced micronuclei in Tradescantia and increased pollen abortion in wild plant species.

  13. Selective accumulation of 147Pm in organism on induction of PCE's micronucleus and SCE of bone marrow cells as well as the chromosome aberrations on fetal liver and spleen

    International Nuclear Information System (INIS)

    Zhu Shoupeng; Zheng Siying; Wang Liuyi; Lu Zhongyan; Yang Shuqin

    1989-01-01

    Study of accumulation peculiarity of 147 Pm showed that I.V. different doses of 147 Pm were the same selectively localized in skeleton and liver. Retention of 147 Pm in skeleton and liver was elevated when the radioactive doses of 147 Pm were increased. At the same time absorption does of 147 Pm radiation was heightened. The ability of 147 Pm to induce sister chromatid exchanges (SCEs) has been investigated by IdU labelling methods. A statistically significant elevation of SCEs was observed after 147 Pm intake.In mice the number of SCEs per cell in bone marrow cells was always higher when the animals were maintained on the doses of 37 Bq/g. The injurious effects of 147 Pm, using PCE's micronucleus rates in bone marrow cells were observed. 147 Pm was dominantly deposited on maternal liver. Deposition of 147 Pm in maternal spleen was about quandrantal of the maternal liver. Studies indicated that maternal contamination of 147 Pm could induced chromosome aberrations in fetal liver and spleen cells. Among the type of aberrations induced by 147 Pm, chromatid breakage were predominant. The incidence of chromosome aberrations on fetal liver cells induced by 147 Pm was higher on fetal spleen cells

  14. Surgical Apgar Score Predicts Post- Laparatomy Complications

    African Journals Online (AJOL)

    calculated Surgical Apgar Scores for 152 patients during a 6-month study ... major postoperative complications and/or death within. 30 days of ... respond to and control hemodynamic changes during a ... abdominal injury (18.42%). Intestinal ...

  15. Budget Scoring: An Impediment to Alternative Financing

    National Research Council Canada - National Science Library

    Summers, Donald E; San Miguel, Joseph G

    2007-01-01

    .... One of the major impediments to using alternative forms of procurement financing for acquiring defense capabilities is in the budgetary treatment, or scoring, of these initiatives by the Congressional Budget Office (CBO...

  16. Film scoring today - Theory, practice and analysis

    OpenAIRE

    Flach, Paula Sophie

    2012-01-01

    This thesis considers film scoring by taking a closer look at the theoretical discourse throughout the last decades, examining current production practice of film music and showcasing a musical analysis of the film Inception (2010).

  17. Climiate Resilience Screening Index and Domain Scores

    Data.gov (United States)

    U.S. Environmental Protection Agency — CRSI and related-domain scores for all 50 states and 3135 counties in the U.S. This dataset is not publicly accessible because: They are already available within the...

  18. Model for predicting the injury severity score.

    Science.gov (United States)

    Hagiwara, Shuichi; Oshima, Kiyohiro; Murata, Masato; Kaneko, Minoru; Aoki, Makoto; Kanbe, Masahiko; Nakamura, Takuro; Ohyama, Yoshio; Tamura, Jun'ichi

    2015-07-01

    To determine the formula that predicts the injury severity score from parameters that are obtained in the emergency department at arrival. We reviewed the medical records of trauma patients who were transferred to the emergency department of Gunma University Hospital between January 2010 and December 2010. The injury severity score, age, mean blood pressure, heart rate, Glasgow coma scale, hemoglobin, hematocrit, red blood cell count, platelet count, fibrinogen, international normalized ratio of prothrombin time, activated partial thromboplastin time, and fibrin degradation products, were examined in those patients on arrival. To determine the formula that predicts the injury severity score, multiple linear regression analysis was carried out. The injury severity score was set as the dependent variable, and the other parameters were set as candidate objective variables. IBM spss Statistics 20 was used for the statistical analysis. Statistical significance was set at P  Watson ratio was 2.200. A formula for predicting the injury severity score in trauma patients was developed with ordinary parameters such as fibrin degradation products and mean blood pressure. This formula is useful because we can predict the injury severity score easily in the emergency department.

  19. Technology Performance Level (TPL) Scoring Tool

    Energy Technology Data Exchange (ETDEWEB)

    Weber, Jochem [National Renewable Energy Lab. (NREL), Golden, CO (United States); Roberts, Jesse D. [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Costello, Ronan [Wave Venture, Penstraze (United Kingdom); Bull, Diana L. [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Babarit, Aurelien [Ecole Centrale de Nantes (France). Lab. of Research in Hydrodynamics, Energetics, and Atmospheric Environment (LHEEA); Neilson, Kim [Ramboll, Copenhagen (Denmark); Bittencourt, Claudio [DNV GL, London (United Kingdom); Kennedy, Ben [Wave Venture, Penstraze (United Kingdom)

    2016-09-01

    Three different ways of combining scores are used in the revised formulation. These are arithmetic mean, geometric mean and multiplication with normalisation. Arithmetic mean is used when combining scores that measure similar attributes, e.g. used for combining costs. The arithmetic mean has the property that it is similar to a logical OR, e.g. when combining costs it does not matter what the individual costs are only what the combined cost is. Geometric mean and Multiplication are used when combining scores that measure disparate attributes. Multiplication is similar to a logical AND, it is used to combine ‘must haves.’ As a result, this method is more punitive than the geometric mean; to get a good score in the combined result it is necessary to have a good score in ALL of the inputs. e.g. the different types of survivability are ‘must haves.’ On balance, the revised TPL is probably less punitive than the previous spreadsheet, multiplication is used sparingly as a method of combining scores. This is in line with the feedback of the Wave Energy Prize judges.

  20. GalaxyDock BP2 score: a hybrid scoring function for accurate protein-ligand docking

    Science.gov (United States)

    Baek, Minkyung; Shin, Woong-Hee; Chung, Hwan Won; Seok, Chaok

    2017-07-01

    Protein-ligand docking is a useful tool for providing atomic-level understanding of protein functions in nature and design principles for artificial ligands or proteins with desired properties. The ability to identify the true binding pose of a ligand to a target protein among numerous possible candidate poses is an essential requirement for successful protein-ligand docking. Many previously developed docking scoring functions were trained to reproduce experimental binding affinities and were also used for scoring binding poses. However, in this study, we developed a new docking scoring function, called GalaxyDock BP2 Score, by directly training the scoring power of binding poses. This function is a hybrid of physics-based, empirical, and knowledge-based score terms that are balanced to strengthen the advantages of each component. The performance of the new scoring function exhibits significant improvement over existing scoring functions in decoy pose discrimination tests. In addition, when the score is used with the GalaxyDock2 protein-ligand docking program, it outperformed other state-of-the-art docking programs in docking tests on the Astex diverse set, the Cross2009 benchmark set, and the Astex non-native set. GalaxyDock BP2 Score and GalaxyDock2 with this score are freely available at http://galaxy.seoklab.org/softwares/galaxydock.html.

  1. MODIFIED ALVARADO SCORING IN ACUTE APPENDICITIS

    Directory of Open Access Journals (Sweden)

    Varadarajan Sujath

    2016-12-01

    Full Text Available BACKGROUND Acute appendicitis is one of the most common surgical emergencies with a lifetime presentation of approximately 1 in 7. Its incidence is 1.5-1.9/1000 in males and females. Surgery for acute appendicitis is based on history, clinical examination and laboratory investigations (e.g. WBC count. Imaging techniques add very little to the efficacy in the diagnosis of appendix. A negative appendicectomy rate of 20-40% has been reported in literature. A difficulty in diagnosis is experienced in very young patients and females of reproductive age. The diagnostic accuracy in assessing acute appendicitis has not improved in spite of rapid advances in management. MATERIALS AND METHODS The modified Alvarado score was applied and assessed for its accuracy in preparation diagnosis of acute appendicitis in 50 patients. The aim of our study is to understand the various presentations of acute appendicitis including the age and gender incidence and the application of the modified Alvarado scoring system in our hospital setup and assessment of the efficacy of the score. RESULTS Our study shows that most involved age group is 3 rd decade with male preponderance. On application of Alvarado score, nausea and vomiting present in 50% and anorexia in 30%, leucocytosis was found in 75% of cases. Sensitivity and specificity of our study were 65% and 40% respectively with positive predictive value of 85% and negative predictive value of 15%. CONCLUSION This study showed that clinical scoring like the Alvarado score can be a cheap and quick tool to apply in emergency departments to rule out acute appendicitis. The implementation of modified Alvarado score is simple and cost effective.

  2. Heart valve surgery: EuroSCORE vs. EuroSCORE II vs. Society of Thoracic Surgeons score

    Directory of Open Access Journals (Sweden)

    Muhammad Sharoz Rabbani

    2014-12-01

    Full Text Available Background This is a validation study comparing the European System for Cardiac Operative Risk Evaluation (EuroSCORE II with the previous additive (AES and logistic EuroSCORE (LES and the Society of Thoracic Surgeons’ (STS risk prediction algorithm, for patients undergoing valve replacement with or without bypass in Pakistan. Patients and Methods Clinical data of 576 patients undergoing valve replacement surgery between 2006 and 2013 were retrospectively collected and individual expected risks of death were calculated by all four risk prediction algorithms. Performance of these risk algorithms was evaluated in terms of discrimination and calibration. Results There were 28 deaths (4.8% among 576 patients, which was lower than the predicted mortality of 5.16%, 6.96% and 4.94% by AES, LES and EuroSCORE II but was higher than 2.13% predicted by STS scoring system. For single and double valve replacement procedures, EuroSCORE II was the best predictor of mortality with highest Hosmer and Lemmeshow test (H-L p value (0.346 to 0.689 and area under the receiver operating characteristic (ROC curve (0.637 to 0.898. For valve plus concomitant coronary artery bypass grafting (CABG patients actual mortality was 1.88%. STS calculator came out to be the best predictor of mortality for this subgroup with H-L p value (0.480 to 0.884 and ROC (0.657 to 0.775. Conclusions For Pakistani population EuroSCORE II is an accurate predictor for individual operative risk in patients undergoing isolated valve surgery, whereas STS performs better in the valve plus CABG group.

  3. WebScore: An Effective Page Scoring Approach for Uncertain Web Social Networks

    Directory of Open Access Journals (Sweden)

    Shaojie Qiao

    2011-10-01

    Full Text Available To effectively score pages with uncertainty in web social networks, we first proposed a new concept called transition probability matrix and formally defined the uncertainty in web social networks. Second, we proposed a hybrid page scoring algorithm, called WebScore, based on the PageRank algorithm and three centrality measures including degree, betweenness, and closeness. Particularly,WebScore takes into a full consideration of the uncertainty of web social networks by computing the transition probability from one page to another. The basic idea ofWebScore is to: (1 integrate uncertainty into PageRank in order to accurately rank pages, and (2 apply the centrality measures to calculate the importance of pages in web social networks. In order to verify the performance of WebScore, we developed a web social network analysis system which can partition web pages into distinct groups and score them in an effective fashion. Finally, we conducted extensive experiments on real data and the results show that WebScore is effective at scoring uncertain pages with less time deficiency than PageRank and centrality measures based page scoring algorithms.

  4. Gambling scores for earthquake predictions and forecasts

    Science.gov (United States)

    Zhuang, Jiancang

    2010-04-01

    This paper presents a new method, namely the gambling score, for scoring the performance earthquake forecasts or predictions. Unlike most other scoring procedures that require a regular scheme of forecast and treat each earthquake equally, regardless their magnitude, this new scoring method compensates the risk that the forecaster has taken. Starting with a certain number of reputation points, once a forecaster makes a prediction or forecast, he is assumed to have betted some points of his reputation. The reference model, which plays the role of the house, determines how many reputation points the forecaster can gain if he succeeds, according to a fair rule, and also takes away the reputation points betted by the forecaster if he loses. This method is also extended to the continuous case of point process models, where the reputation points betted by the forecaster become a continuous mass on the space-time-magnitude range of interest. We also calculate the upper bound of the gambling score when the true model is a renewal process, the stress release model or the ETAS model and when the reference model is the Poisson model.

  5. Quality scores for 32,000 genomes

    DEFF Research Database (Denmark)

    Land, Miriam L.; Hyatt, Doug; Jun, Se-Ran

    2014-01-01

    Background More than 80% of the microbial genomes in GenBank are of ‘draft’ quality (12,553 draft vs. 2,679 finished, as of October, 2013). We have examined all the microbial DNA sequences available for complete, draft, and Sequence Read Archive genomes in GenBank as well as three other major...... public databases, and assigned quality scores for more than 30,000 prokaryotic genome sequences. Results Scores were assigned using four categories: the completeness of the assembly, the presence of full-length rRNA genes, tRNA composition and the presence of a set of 102 conserved genes in prokaryotes....... Most (~88%) of the genomes had quality scores of 0.8 or better and can be safely used for standard comparative genomics analysis. We compared genomes across factors that may influence the score. We found that although sequencing depth coverage of over 100x did not ensure a better score, sequencing read...

  6. An ultrasound score for knee osteoarthritis

    DEFF Research Database (Denmark)

    Riecke, B F; Christensen, R.; Torp-Pedersen, S

    2014-01-01

    OBJECTIVE: To develop standardized musculoskeletal ultrasound (MUS) procedures and scoring for detecting knee osteoarthritis (OA) and test the MUS score's ability to discern various degrees of knee OA, in comparison with plain radiography and the 'Knee injury and Osteoarthritis Outcome Score' (KOOS......) domains as comparators. METHOD: A cross-sectional study of MUS examinations in 45 patients with knee OA. Validity, reliability, and reproducibility were evaluated. RESULTS: MUS examination for knee OA consists of five separate domains assessing (1) predominantly morphological changes in the medial...... coefficients ranging from 0.75 to 0.97 for the five domains. Construct validity was confirmed with statistically significant correlation coefficients (0.47-0.81, P knee OA. In comparison with standing radiographs...

  7. Assigning Numerical Scores to Linguistic Expressions

    Directory of Open Access Journals (Sweden)

    María Jesús Campión

    2017-07-01

    Full Text Available In this paper, we study different methods of scoring linguistic expressions defined on a finite set, in the search for a linear order that ranks all those possible expressions. Among them, particular attention is paid to the canonical extension, and its representability through distances in a graph plus some suitable penalization of imprecision. The relationship between this setting and the classical problems of numerical representability of orderings, as well as extension of orderings from a set to a superset is also explored. Finally, aggregation procedures of qualitative rankings and scorings are also analyzed.

  8. What Do Test Scores Really Mean? A Latent Class Analysis of Danish Test Score Performance

    DEFF Research Database (Denmark)

    Munk, Martin D.; McIntosh, James

    2014-01-01

    Latent class Poisson count models are used to analyze a sample of Danish test score results from a cohort of individuals born in 1954-55, tested in 1968, and followed until 2011. The procedure takes account of unobservable effects as well as excessive zeros in the data. We show that the test scores...... of intelligence explain a significant proportion of the variation in test scores. This adds to the complexity of interpreting test scores and suggests that school culture and possible incentive problems make it more di¢ cult to understand what the tests measure....

  9. NCACO-score: An effective main-chain dependent scoring function for structure modeling

    Directory of Open Access Journals (Sweden)

    Dong Xiaoxi

    2011-05-01

    Full Text Available Abstract Background Development of effective scoring functions is a critical component to the success of protein structure modeling. Previously, many efforts have been dedicated to the development of scoring functions. Despite these efforts, development of an effective scoring function that can achieve both good accuracy and fast speed still presents a grand challenge. Results Based on a coarse-grained representation of a protein structure by using only four main-chain atoms: N, Cα, C and O, we develop a knowledge-based scoring function, called NCACO-score, that integrates different structural information to rapidly model protein structure from sequence. In testing on the Decoys'R'Us sets, we found that NCACO-score can effectively recognize native conformers from their decoys. Furthermore, we demonstrate that NCACO-score can effectively guide fragment assembly for protein structure prediction, which has achieved a good performance in building the structure models for hard targets from CASP8 in terms of both accuracy and speed. Conclusions Although NCACO-score is developed based on a coarse-grained model, it is able to discriminate native conformers from decoy conformers with high accuracy. NCACO is a very effective scoring function for structure modeling.

  10. Algorithm improvement program nuclide identification algorithm scoring criteria and scoring application.

    Energy Technology Data Exchange (ETDEWEB)

    Enghauser, Michael [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)

    2016-02-01

    The goal of the Domestic Nuclear Detection Office (DNDO) Algorithm Improvement Program (AIP) is to facilitate gamma-radiation detector nuclide identification algorithm development, improvement, and validation. Accordingly, scoring criteria have been developed to objectively assess the performance of nuclide identification algorithms. In addition, a Microsoft Excel spreadsheet application for automated nuclide identification scoring has been developed. This report provides an overview of the equations, nuclide weighting factors, nuclide equivalencies, and configuration weighting factors used by the application for scoring nuclide identification algorithm performance. Furthermore, this report presents a general overview of the nuclide identification algorithm scoring application including illustrative examples.

  11. Graphistrength© C100 MultiWalled Carbon Nanotubes (MWCNT): thirteen-week inhalation toxicity study in rats with 13- and 52-week recovery periods combined with comet and micronucleus assays

    International Nuclear Information System (INIS)

    Régnier, Jean-François; Pothmann-Krings, Daniela; Simar, Sophie; Dony, Eva; Net, Jean-Loïc Le; Beausoleil, Julien

    2017-01-01

    Graphistrength© C100 provides superior electrical and mechanical properties for various applications and is one of the industrial MWCNT referenced in the OECD sponsorship program for the safety testing of nanomaterials. Graphistrength© C100 is formed of MWCNT (ca. 12 walls, outer mean diameter ca. 12 nm, length ca. 1 µm) agglomerated in particles with a granulometry centered on 400 µm. A general feature of MWCNT after inhalation or intratracheal exposures is the induction of an inflammatory reaction in the lungs sometimes associated with local genotoxic effects. Most of the in vitro and in vivo genotoxicity data available on Graphistrength© C100 are negative. However, a weak DNA damage activity in the in vitro and in vivo FPG-modified Comet assays and a weak clastogenic effect in the in vitr o micronucleus test were reported. After investigating different parameters for the aerosol generation, male and female Wistar rats were exposed by nose-only inhalation (6h/day, 5d/week) to target concentrations of 0.05, 0.25 and 5.0 mg/m 3 air of a respirable aerosol (MMAD < 3 µm) and sacrificed immediately after 4 and 13 weeks of exposure and 13 and 52 weeks of recovery after the 13-week exposure. Clinical, biological and histological evaluations were performed according to the OECD TG 413. Broncho-alveolar lavage fluid (BALF) was collected and analysed for cytokines and inflammatory parameters. Immediately after 13 weeks of exposure, chromosomal aberrations in the bone marrow cells of males and females were evaluated by the micronucleus test (OECD TG 474) and DNA damage in the lung, kidney and liver cells of males were assessed by both the standard and the human 8-oxoguanine DNA N-glycosylase 1 (hOGG1)-modified comet assay (OECD TG 489). Concentration-related deposition of black particles (MWCNT) was observed in lungs. At all sacrifice periods, an inflammatory lung reaction was observed in rats exposed to 5.0 mg/m 3 associated with changes in the differential white

  12. Graphistrength© C100 MultiWalled Carbon Nanotubes (MWCNT): thirteen-week inhalation toxicity study in rats with 13- and 52-week recovery periods combined with comet and micronucleus assays

    Science.gov (United States)

    Régnier, Jean-François; Pothmann-Krings, Daniela; Simar, Sophie; Dony, Eva; Le Net, Jean-Loïc; Beausoleil, Julien

    2017-06-01

    Graphistrength© C100 provides superior electrical and mechanical properties for various applications and is one of the industrial MWCNT referenced in the OECD sponsorship program for the safety testing of nanomaterials. Graphistrength© C100 is formed of MWCNT (ca. 12 walls, outer mean diameter ca. 12 nm, length ca. 1 µm) agglomerated in particles with a granulometry centered on 400 µm. A general feature of MWCNT after inhalation or intratracheal exposures is the induction of an inflammatory reaction in the lungs sometimes associated with local genotoxic effects. Most of the in vitro and in vivo genotoxicity data available on Graphistrength© C100 are negative. However, a weak DNA damage activity in the in vitro and in vivo FPG-modified Comet assays and a weak clastogenic effect in the in vitro micronucleus test were reported. After investigating different parameters for the aerosol generation, male and female Wistar rats were exposed by nose-only inhalation (6h/day, 5d/week) to target concentrations of 0.05, 0.25 and 5.0 mg/m3 air of a respirable aerosol (MMAD males and females were evaluated by the micronucleus test (OECD TG 474) and DNA damage in the lung, kidney and liver cells of males were assessed by both the standard and the human 8-oxoguanine DNA N-glycosylase 1 (hOGG1)-modified comet assay (OECD TG 489). Concentration-related deposition of black particles (MWCNT) was observed in lungs. At all sacrifice periods, an inflammatory lung reaction was observed in rats exposed to 5.0 mg/m3 associated with changes in the differential white blood cells counts. The lung inflammation was characterized by changes in the cytological, biochemical and cytokine parameters of the BALF, an increase of the lung weight, an interstitial inflammation mainly around the alveolar ducts at the bronchiole-alveolar junction and a cell hypertrophy/hyperplasia in the terminal and respiratory bronchioles. The slight changes in BALF parameters observed at 0.25 mg/m3 recovered after

  13. Commentary: critical questions, misconceptions and a road map for improving the use of the lymphocyte cytokinesis-block micronucleus assay for in vivo biomonitoring of human exposure to genotoxic chemicals-a HUMN project perspective.

    Science.gov (United States)

    Kirsch-Volders, Micheline; Bonassi, Stefano; Knasmueller, Siegfried; Holland, Nina; Bolognesi, Claudia; Fenech, Michael F

    2014-01-01

    The lymphocyte cytokinesis-block micronucleus (CBMN) assay has been applied in hundreds of in vivo biomonitoring studies of humans exposed to genotoxic chemicals because it allows the measurement of both structural and numerical chromosome aberrations. The CBMN cytome assay version which, apart from measuring micronuclei (MN) already present in cells in vivo or expressed ex vivo, also includes measurement of nucleoplasmic bridges (NPB), nuclear buds (NBUD), necrosis and apoptosis, is also increasingly being used in such studies. Because of the numerous published studies there is now a need to re-evaluate the use of MN and other biomarkers within the lymphocyte CBMN cytome assay as quantitative indicators of exposure to chemical genotoxins and the genetic hazard this may cause. This review has identified some important misconceptions as well as knowledge gaps that need to be addressed to make further progress in the proper application of this promising technique and enable its full potential to be realised. The HUMN project consortium recommends a three pronged approach to further improve the knowledge base and application of the lymphocyte CBMN cytome assay to measure DNA damage in humans exposed to chemical genotoxins: (i) a series of systematic reviews, one for each class of chemical genotoxins, of studies which have investigated the association of in vivo exposure in humans with MN, NPB and NBUD induction in lymphocytes; (ii) a comprehensive analysis of the literature to obtain new insights on the potential mechanisms by which different classes of chemicals may induce MN, NPB and NBUD in vitro and in vivo and (iii) investigation of the potential advantages of using the lymphocyte CBMN cytome assay in conjunction with other promising complementary DNA damage diagnostics to obtain an even more complete assessment of the DNA damage profile induced by in vivo exposure to chemical genotoxins in humans. Copyright © 2014 The Authors. Published by Elsevier B.V. All

  14. Scoring ultrasound synovitis in rheumatoid arthritis

    DEFF Research Database (Denmark)

    D'Agostino, Maria-Antonietta; Terslev, Lene; Aegerter, Philippe

    2017-01-01

    OBJECTIVES: To develop a consensus-based ultrasound (US) definition and quantification system for synovitis in rheumatoid arthritis (RA). METHODS: A multistep, iterative approach was used to: (1) evaluate the baseline agreement on defining and scoring synovitis according to the usual practice...

  15. Multilevel Analysis of Student Civics Knowledge Scores

    Science.gov (United States)

    Gregory, Chris; Miyazaki, Yasuo

    2018-01-01

    Compositional effects of scholarly culture classroom/school climate on civic knowledge scores of 9th graders in the United States were examined using the International Association for the Evaluation of Educational Achievement (IEA) 1999 Civic Education Study data. Following Evans et al. (2010, 2014), we conceived that the number of books at home,…

  16. Normalization of the Psychometric Hepatic Encephalopathy score ...

    African Journals Online (AJOL)

    2016-05-09

    May 9, 2016 ... influenced by age, education levels, and gender.[5] Till date, the PHES ... and death. MHE also increases the risk of development ... large circles beginning from each row on the left and working to the right. The test score is the ...

  17. SCORE - Sounding-rocket Coronagraphic Experiment

    Science.gov (United States)

    Fineschi, Silvano; Moses, Dan; Romoli, Marco

    The Sounding-rocket Coronagraphic Experiment - SCORE - is a The Sounding-rocket Coronagraphic Experiment - SCORE - is a coronagraph for multi-wavelength imaging of the coronal Lyman-alpha lines, HeII 30.4 nm and HI 121.6 nm, and for the broad.band visible-light emission of the polarized K-corona. SCORE has flown successfully in 2009 acquiring the first images of the HeII line-emission from the extended corona. The simultaneous observation of the coronal Lyman-alpha HI 121.6 nm, has allowed the first determination of the absolute helium abundance in the extended corona. This presentation will describe the lesson learned from the first flight and will illustrate the preparations and the science perspectives for the second re-flight approved by NASA and scheduled for 2016. The SCORE optical design is flexible enough to be able to accommodate different experimental configurations with minor modifications. This presentation will describe one of such configurations that could include a polarimeter for the observation the expected Hanle effect in the coronal Lyman-alpha HI line. The linear polarization by resonance scattering of coronal permitted line-emission in the ultraviolet (UV) can be modified by magnetic fields through the Hanle effect. Thus, space-based UV spectro-polarimetry would provide an additional new tool for the diagnostics of coronal magnetism.

  18. Effects of heterogeneity on bank efficiency scores

    NARCIS (Netherlands)

    Bos, J. W. B.; Koetter, M.; Kolari, J. W.; Kool, C. J. M.

    2009-01-01

    Bank efficiency estimates often serve as a proxy of managerial skill since they quantify sub-optimal production choices. But such deviations can also be due to omitted systematic differences among banks. In this study, we examine the effects of heterogeneity on bank efficiency scores. We compare

  19. Correlation between International Prostate Symptom Score and ...

    African Journals Online (AJOL)

    2016-07-23

    Jul 23, 2016 ... International Prostate Symptom Score (IPSS) and uroflowmetry in patients with lower urinary tract symptoms-benign prostatic ... cause of bladder outlet obstruction (BOO) in the male geriatric population.[1] ... age and results in LUTS in about 10% of elderly men.[1]. BPH causes morbidity through the urinary ...

  20. Local Observed-Score Kernel Equating

    Science.gov (United States)

    Wiberg, Marie; van der Linden, Wim J.; von Davier, Alina A.

    2014-01-01

    Three local observed-score kernel equating methods that integrate methods from the local equating and kernel equating frameworks are proposed. The new methods were compared with their earlier counterparts with respect to such measures as bias--as defined by Lord's criterion of equity--and percent relative error. The local kernel item response…

  1. The scoring of movements in sleep.

    Science.gov (United States)

    Walters, Arthur S; Lavigne, Gilles; Hening, Wayne; Picchietti, Daniel L; Allen, Richard P; Chokroverty, Sudhansu; Kushida, Clete A; Bliwise, Donald L; Mahowald, Mark W; Schenck, Carlos H; Ancoli-Israel, Sonia

    2007-03-15

    The International Classification of Sleep Disorders (ICSD-2) has separated sleep-related movement disorders into simple, repetitive movement disorders (such as periodic limb movements in sleep [PLMS], sleep bruxism, and rhythmic movement disorder) and parasomnias (such as REM sleep behavior disorder and disorders of partial arousal, e.g., sleep walking, confusional arousals, night terrors). Many of the parasomnias are characterized by complex behaviors in sleep that appear purposeful, goal directed and voluntary but are outside the conscious awareness of the individual and therefore inappropriate. All of the sleep-related movement disorders described here have specific polysomnographic findings. For the purposes of developing and/or revising specifications and polysomnographic scoring rules, the AASM Scoring Manual Task Force on Movements in Sleep reviewed background literature and executed evidence grading of 81 relevant articles obtained by a literature search of published articles between 1966 and 2004. Subsequent evidence grading identified limited evidence for reliability and/or validity for polysomnographic scoring criteria for periodic limb movements in sleep, REM sleep behavior disorder, and sleep bruxism. Published scoring criteria for rhythmic movement disorder, excessive fragmentary myoclonus, and hypnagogic foot tremor/alternating leg muscle activation were empirical and based on descriptive studies. The literature review disclosed no published evidence defining clinical consequences of excessive fragmentary myoclonus or hypnagogic foot tremor/alternating leg muscle activation. Because of limited or absent evidence for reliability and/or validity, a standardized RAND/UCLA consensus process was employed for recommendation of specific rules for the scoring of sleep-associated movements.

  2. Validation of dengue infection severity score

    Directory of Open Access Journals (Sweden)

    Pongpan S

    2014-03-01

    Full Text Available Surangrat Pongpan,1,2 Jayanton Patumanond,3 Apichart Wisitwong,4 Chamaiporn Tawichasri,5 Sirianong Namwongprom1,6 1Clinical Epidemiology Program, Faculty of Medicine, Chiang Mai University, Chiang Mai, Thailand; 2Department of Occupational Medicine, Phrae Hospital, Phrae, Thailand; 3Clinical Epidemiology Program, Faculty of Medicine, Thammasat University, Bangkok, Thailand; 4Department of Social Medicine, Sawanpracharak Hospital, Nakorn Sawan, Thailand; 5Clinical Epidemiology Society at Chiang Mai, Chiang Mai, Thailand; 6Department of Radiology, Faculty of Medicine, Chiang Mai University, Chiang Mai, Thailand Objective: To validate a simple scoring system to classify dengue viral infection severity to patients in different settings. Methods: The developed scoring system derived from 777 patients from three tertiary-care hospitals was applied to 400 patients in the validation data obtained from another three tertiary-care hospitals. Percentage of correct classification, underestimation, and overestimation was compared. The score discriminative performance in the two datasets was compared by analysis of areas under the receiver operating characteristic curves. Results: Patients in the validation data were different from those in the development data in some aspects. In the validation data, classifying patients into three severity levels (dengue fever, dengue hemorrhagic fever, and dengue shock syndrome yielded 50.8% correct prediction (versus 60.7% in the development data, with clinically acceptable underestimation (18.6% versus 25.7% and overestimation (30.8% versus 13.5%. Despite the difference in predictive performances between the validation and the development data, the overall prediction of the scoring system is considered high. Conclusion: The developed severity score may be applied to classify patients with dengue viral infection into three severity levels with clinically acceptable under- or overestimation. Its impact when used in routine

  3. Development of the Crohn's disease digestive damage score, the Lémann score

    DEFF Research Database (Denmark)

    Pariente, Benjamin; Cosnes, Jacques; Danese, Silvio

    2011-01-01

    is to outline the methods to develop an instrument that can measure cumulative bowel damage. The project is being conducted by the International Program to develop New Indexes in Crohn's disease (IPNIC) group. This instrument, called the Crohn's Disease Digestive Damage Score (the Lémann score), should take...

  4. Relationship between Students' Scores on Research Methods and Statistics, and Undergraduate Project Scores

    Science.gov (United States)

    Ossai, Peter Agbadobi Uloku

    2016-01-01

    This study examined the relationship between students' scores on Research Methods and statistics, and undergraduate project at the final year. The purpose was to find out whether students matched knowledge of research with project-writing skill. The study adopted an expost facto correlational design. Scores on Research Methods and Statistics for…

  5. The RIPASA score for the diagnosis of acute appendicitis: A comparison with the modified Alvarado score.

    Science.gov (United States)

    Díaz-Barrientos, C Z; Aquino-González, A; Heredia-Montaño, M; Navarro-Tovar, F; Pineda-Espinosa, M A; Espinosa de Santillana, I A

    2018-02-06

    Acute appendicitis is the first cause of surgical emergencies. It is still a difficult diagnosis to make, especially in young persons, the elderly, and in reproductive-age women, in whom a series of inflammatory conditions can have signs and symptoms similar to those of acute appendicitis. Different scoring systems have been created to increase diagnostic accuracy, and they are inexpensive, noninvasive, and easy to use and reproduce. The modified Alvarado score is probably the most widely used and accepted in emergency services worldwide. On the other hand, the RIPASA score was formulated in 2010 and has greater sensitivity and specificity. There are very few studies conducted in Mexico that compare the different scoring systems for appendicitis. The aim of our article was to compare the modified Alvarado score and the RIPASA score in the diagnosis of patients with abdominal pain and suspected acute appendicitis. An observational, analytic, and prolective study was conducted within the time frame of July 2002 and February 2014 at the Hospital Universitario de Puebla. The questionnaires used for the evaluation process were applied to the patients suspected of having appendicitis. The RIPASA score with 8.5 as the optimal cutoff value: ROC curve (area .595), sensitivity (93.3%), specificity (8.3%), PPV (91.8%), NPV (10.1%). Modified Alvarado score with 6 as the optimal cutoff value: ROC curve (area .719), sensitivity (75%), specificity (41.6%), PPV (93.7%), NPV (12.5%). The RIPASA score showed no advantages over the modified Alvarado score when applied to patients presenting with suspected acute appendicitis. Copyright © 2018 Asociación Mexicana de Gastroenterología. Publicado por Masson Doyma México S.A. All rights reserved.

  6. Lower bounds to the reliabilities of factor score estimators

    NARCIS (Netherlands)

    Hessen, D.J.

    2017-01-01

    Under the general common factor model, the reliabilities of factor score estimators might be of more interest than the reliability of the total score (the unweighted sum of item scores). In this paper, lower bounds to the reliabilities of Thurstone’s factor score estimators, Bartlett’s factor score

  7. 2-Aminoanthracene, 5-fluorouracil, colchicine, benzo[a]pyrene, cadmium chloride and cytosine arabinoside tested in the in vitro mammalian cell micronucleus test (MNvit) in Chinese hamster ovary (CHO) cells at Covance Laboratories, Harrogate UK in support of OECD draft Test Guideline 487.

    Science.gov (United States)

    Whitwell, James; Fowler, Paul; Allars, Sarah; Jenner, Karen; Lloyd, Melvyn; Wood, Debbie; Smith, Katie; Young, Jamie; Jeffrey, Laura; Kirkland, David

    2010-10-29

    The reference genotoxic agents 2-aminoanthracene (a metabolism dependent weak clastogen), 5-fluorouracil (a nucleoside analogue, characterised by a steep dose response profile), colchicine (an aneugen that inhibits tubulin polymerisation), benzo[a]pyrene (a polycyclic aromatic hydrocarbon requiring metabolic activation), cadmium chloride (an inorganic carcinogen), and cytosine arabinoside (a nucleoside analogue that inhibits the gap-filling step of excision repair) were tested in the in vitro micronucleus assay using the Chinese hamster ovary (CHO) cell line at Covance Laboratories, Harrogate, UK. All chemicals were treated in the absence and presence of cytokinesis block (via addition of cytochalasin B) with this work forming part of a collaborative evaluation of the toxicity measures recommended in the draft OECD Test Guideline 487 on the In vitro Mammalian Cell Micronucleus Test (MNvit). The toxicity measures used, detecting a possible combination of both cytostasis and cell death (though not cell death directly), were relative population doubling, relative increase in cell counts and relative cell counts for treatments in the absence of cytokinesis block, and replication index in the presence of cytokinesis block. All of the chemicals tested either gave marked positive increases in the percentage of micronucleated cells with and without cytokinesis block, or did not induce micronuclei at concentrations giving approximately 50-60% toxicity (cytostasis and cell death) or less by all of the toxicity measures used. The outcome from this series of tests supports the use of relative increase in cell counts and relative population doubling, as well as relative cell counts, as appropriate measures of cytotoxicity for the non-cytokinesis blocked in vitro micronucleus assay. Copyright © 2010 Elsevier B.V. All rights reserved.

  8. Scoring Rules for Subjective Probability Distributions

    DEFF Research Database (Denmark)

    Harrison, Glenn W.; Martínez-Correa, Jimmy; Swarthout, J. Todd

    The theoretical literature has a rich characterization of scoring rules for eliciting the subjective beliefs that an individual has for continuous events, but under the restrictive assumption of risk neutrality. It is well known that risk aversion can dramatically affect the incentives to correctly...... report the true subjective probability of a binary event, even under Subjective Expected Utility. To address this one can “calibrate” inferences about true subjective probabilities from elicited subjective probabilities over binary events, recognizing the incentives that risk averse agents have...... to distort reports. We characterize the comparable implications of the general case of a risk averse agent when facing a popular scoring rule over continuous events, and find that these concerns do not apply with anything like the same force. For empirically plausible levels of risk aversion, one can...

  9. Shower reconstruction in TUNKA-HiSCORE

    Energy Technology Data Exchange (ETDEWEB)

    Porelli, Andrea; Wischnewski, Ralf [DESY-Zeuthen, Platanenallee 6, 15738 Zeuthen (Germany)

    2015-07-01

    The Tunka-HiSCORE detector is a non-imaging wide-angle EAS cherenkov array designed as an alternative technology for gamma-ray physics above 10 TeV and to study spectrum and composition of cosmic rays above 100 TeV. An engineering array with nine stations (HiS-9) has been deployed in October 2013 on the site of the Tunka experiment in Russia. In November 2014, 20 more HiSCORE stations have been installed, covering a total array area of 0.24 square-km. We describe the detector setup, the role of precision time measurement, and give results from the innovative WhiteRabbit time synchronization technology. Results of air shower reconstruction are presented and compared with MC simulations, for both the HiS-9 and the HiS-29 detector arrays.

  10. Credit scoring analysis using kernel discriminant

    Science.gov (United States)

    Widiharih, T.; Mukid, M. A.; Mustafid

    2018-05-01

    Credit scoring model is an important tool for reducing the risk of wrong decisions when granting credit facilities to applicants. This paper investigate the performance of kernel discriminant model in assessing customer credit risk. Kernel discriminant analysis is a non- parametric method which means that it does not require any assumptions about the probability distribution of the input. The main ingredient is a kernel that allows an efficient computation of Fisher discriminant. We use several kernel such as normal, epanechnikov, biweight, and triweight. The models accuracy was compared each other using data from a financial institution in Indonesia. The results show that kernel discriminant can be an alternative method that can be used to determine who is eligible for a credit loan. In the data we use, it shows that a normal kernel is relevant to be selected for credit scoring using kernel discriminant model. Sensitivity and specificity reach to 0.5556 and 0.5488 respectively.

  11. Nursing Activities Score and Acute Kidney Injury

    Directory of Open Access Journals (Sweden)

    Filipe Utuari de Andrade Coelho

    Full Text Available ABSTRACT Objective: to evaluate the nursing workload in intensive care patients with acute kidney injury (AKI. Method: A quantitative study, conducted in an intensive care unit, from April to August of 2015. The Nursing Activities Score (NAS and Kidney Disease Improving Global Outcomes (KDIGO were used to measure nursing workload and to classify the stage of AKI, respectively. Results: A total of 190 patients were included. Patients who developed AKI (44.2% had higher NAS when compared to those without AKI (43.7% vs 40.7%, p <0.001. Patients with stage 1, 2 and 3 AKI showed higher NAS than those without AKI. A relationship was identified between stage 2 and 3 with those without AKI (p = 0.002 and p <0.001. Conclusion: The NAS was associated with the presence of AKI, the score increased with the progression of the stages, and it was associated with AKI, stage 2 and 3.

  12. Psychometric properties of the Cumulated Ambulation Score

    DEFF Research Database (Denmark)

    Ferriero, Giorgio; Kristensen, Morten T; Invernizzi, Marco

    2018-01-01

    INTRODUCTION: In the geriatric population, independent mobility is a key factor in determining readiness for discharge following acute hospitalization. The Cumulated Ambulation Score (CAS) is a potentially valuable score that allows day-to-day measurements of basic mobility. The CAS was developed...... and validated in older patients with hip fracture as an early postoperative predictor of short-term outcome, but it is also used to assess geriatric in-patients with acute medical illness. Despite the fast- accumulating literature on the CAS, to date no systematic review synthesizing its psychometric properties....... Of 49 studies identified, 17 examined the psychometric properties of the CAS. EVIDENCE SYNTHESIS: Most papers dealt with patients after hip fracture surgery, and only 4 studies assessed the CAS psychometric characteristics also in geriatric in-patients with acute medical illness. Two versions of CAS...

  13. Multi-color CD34⁺ progenitor-focused flow cytometric assay in evaluation of myelodysplastic syndromes in patients with post cancer therapy cytopenia.

    Science.gov (United States)

    Tang, Guilin; Jorgensen, L Jeffrey; Zhou, Yi; Hu, Ying; Kersh, Marian; Garcia-Manero, Guillermo; Medeiros, L Jeffrey; Wang, Sa A

    2012-08-01

    Bone marrow assessment for myelodysplastic syndrome (MDS) in a patient who develops cytopenia(s) following cancer therapy is challenging. With recent advances in multi-color flow cytometry immunophenotypic analysis, a CD34(+) progenitor-focused 7-color assay was developed and tested in this clinical setting. This assay was first performed in 73 MDS patients and 53 non-MDS patients (developmental set). A number of immunophenotypic changes were differentially observed in these two groups. Based on the sensitivity, specificity and reproducibility, a core panel of markers was selected for final assessment that included increased total CD34(+) myeloblasts; decreased stage I hematogones; altered CD45/side scatter; altered expression of CD13, CD33, CD34, CD38, CD117, and CD123; aberrant expression of lymphoid or mature myelomonocytic antigens on CD34(+) myeloblasts; and several marked alterations in maturing myelomonocytic cells. The data were translated into a simplified scoring system which was then used in 120 patients with cytopenia(s) secondary to cancer therapy over a 2-year period (validation set). With a median follow-up of 11 months, this assay demonstrated 89% sensitivity, 94% specificity, and 92% accuracy in establishing or excluding a diagnosis of MDS. Copyright © 2012 Elsevier Ltd. All rights reserved.

  14. PhishScore: Hacking Phishers' Minds

    OpenAIRE

    Marchal, Samuel; François, Jérôme; State, Radu; Engel, Thomas

    2014-01-01

    Despite the growth of prevention techniques, phishing remains an important threat since the principal countermeasures in use are still based on reactive URL blacklisting. This technique is inefficient due to the short lifetime of phishing Web sites, making recent approaches relying on real-time or proactive phishing URLs detection techniques more appropriate. In this paper we introduce PhishScore, an automated real-time phishing detection system. We observed that phishing URLs usually have fe...

  15. Credit Scoring Problem Based on Regression Analysis

    OpenAIRE

    Khassawneh, Bashar Suhil Jad Allah

    2014-01-01

    ABSTRACT: This thesis provides an explanatory introduction to the regression models of data mining and contains basic definitions of key terms in the linear, multiple and logistic regression models. Meanwhile, the aim of this study is to illustrate fitting models for the credit scoring problem using simple linear, multiple linear and logistic regression models and also to analyze the found model functions by statistical tools. Keywords: Data mining, linear regression, logistic regression....

  16. Fingerprint Recognition Using Minutia Score Matching

    OpenAIRE

    J, Ravi.; Raja, K. B.; R, Venugopal. K.

    2010-01-01

    The popular Biometric used to authenticate a person is Fingerprint which is unique and permanent throughout a person’s life. A minutia matching is widely used for fingerprint recognition and can be classified as ridge ending and ridge bifurcation. In this paper we projected Fingerprint Recognition using Minutia Score Matching method (FRMSM). For Fingerprint thinning, the Block Filter is used, which scans the image at the boundary to preserves the quality of the image and extract the minutiae ...

  17. Gender, Stereotype Threat and Mathematics Test Scores

    OpenAIRE

    Ming Tsui; Xiao Y. Xu; Edmond Venator

    2011-01-01

    Problem statement: Stereotype threat has repeatedly been shown to depress womens scores on difficult math tests. An attempt to replicate these findings in China found no support for the stereotype threat hypothesis. Our math test was characterized as being personally important for the student participants, an atypical condition in most stereotype threat laboratory research. Approach: To evaluate the effects of this personal demand, we conducted three experiments. Results: ...

  18. MODELING CREDIT RISK THROUGH CREDIT SCORING

    OpenAIRE

    Adrian Cantemir CALIN; Oana Cristina POPOVICI

    2014-01-01

    Credit risk governs all financial transactions and it is defined as the risk of suffering a loss due to certain shifts in the credit quality of a counterpart. Credit risk literature gravitates around two main modeling approaches: the structural approach and the reduced form approach. In addition to these perspectives, credit risk assessment has been conducted through a series of techniques such as credit scoring models, which form the traditional approach. This paper examines the evolution of...

  19. Superior cold recycling : The score project

    OpenAIRE

    LESUEUR, D; POTTI, JJ; SOUTHWELL, C; WALTER, J; CRUZ, M; DELFOSSE, F; ECKMANN, B; FIEDLER, J; RACEK, I; SIMONSSON, B; PLACIN, F; SERRANO, J; RUIZ, A; KALAAJI, A; ATTANE, P

    2004-01-01

    In order to develop Environmentally Friendly Construction Technologies (EFCT) and as part of the 5th Framework Program of Research and Development, the European Community has decided to finance a research project on cold recycling, entitled SCORE "Superior COld REcycling based on benefits of bituminous microemulsions and foamed bitumen. A EFCT system for the rehabilitation and the maintenance of roads". This research project gathers organizations from all over Europe, from industrial partners...

  20. North Korean refugee doctors' preliminary examination scores

    Directory of Open Access Journals (Sweden)

    Sung Uk Chae

    2016-12-01

    Full Text Available Purpose Although there have been studies emphasizing the re-education of North Korean (NK doctors for post-unification of the Korean Peninsula, study on the content and scope of such re-education has yet to be conducted. Researchers intended to set the content and scope of re-education by a comparative analysis for the scores of the preliminary examination, which is comparable to the Korean Medical Licensing Examination (KMLE. Methods The scores of the first and second preliminary exams were analyzed by subject using the Wilcoxon signed rank test. The passing status of the group of NK doctors for KMLE in recent 3 years were investigated. The multiple-choice-question (MCQ items of which difficulty indexes of NK doctors were lower than those of South Korean (SK medical students by two times of the standard deviation of the scores of SK medical students were selected to investigate the relevant reasons. Results The average scores of nearly all subjects were improved in the second exam compared with the first exam. The passing rate of the group of NK doctors was 75%. The number of MCQ items of which difficulty indexes of NK doctors were lower than those of SK medical students was 51 (6.38%. NK doctors’ lack of understandings for Diagnostic Techniques and Procedures, Therapeutics, Prenatal Care, and Managed Care Programs was suggested as the possible reason. Conclusion The education of integrated courses focusing on Diagnostic Techniques and Procedures and Therapeutics, and apprenticeship-style training for clinical practice of core subjects are needed. Special lectures on the Preventive Medicine are likely to be required also.

  1. What do educational test scores really measure?

    DEFF Research Database (Denmark)

    McIntosh, James; D. Munk, Martin

    Latent class Poisson count models are used to analyze a sample of Danish test score results from a cohort of individuals born in 1954-55 and tested in 1968. The procedure takes account of unobservable effects as well as excessive zeros in the data. The bulk of unobservable effects are uncorrelate......, and possible incentive problems make it more difficult to elicit true values of what the tests measure....

  2. Wearable PPG sensor based alertness scoring system.

    Science.gov (United States)

    Dey, Jishnu; Bhowmik, Tanmoy; Sahoo, Saswata; Tiwari, Vijay Narayan

    2017-07-01

    Quantifying mental alertness in today's world is important as it enables the person to adopt lifestyle changes for better work efficiency. Miniaturized sensors in wearable devices have facilitated detection/monitoring of mental alertness. Photoplethysmography (PPG) sensors through Heart Rate Variability (HRV) offer one such opportunity by providing information about one's daily alertness levels without requiring any manual interference from the user. In this paper, a smartwatch based alertness estimation system is proposed. Data collected from PPG sensor of smartwatch is processed and fed to machine learning based model to get a continuous alertness score. Utility functions are designed based on statistical analysis to give a quality score on different stages of alertness such as awake, long sleep and short duration power nap. An intelligent data collection approach is proposed in collaboration with the motion sensor in the smartwatch to reduce battery drainage. Overall, our proposed wearable based system provides a detailed analysis of alertness over a period in a systematic and optimized manner. We were able to achieve an accuracy of 80.1% for sleep/awake classification along with alertness score. This opens up the possibility for quantifying alertness levels using a single PPG sensor for better management of health related activities including sleep.

  3. High throughput sample processing and automated scoring

    Directory of Open Access Journals (Sweden)

    Gunnar eBrunborg

    2014-10-01

    Full Text Available The comet assay is a sensitive and versatile method for assessing DNA damage in cells. In the traditional version of the assay, there are many manual steps involved and few samples can be treated in one experiment. High throughput modifications have been developed during recent years, and they are reviewed and discussed. These modifications include accelerated scoring of comets; other important elements that have been studied and adapted to high throughput are cultivation and manipulation of cells or tissues before and after exposure, and freezing of treated samples until comet analysis and scoring. High throughput methods save time and money but they are useful also for other reasons: large-scale experiments may be performed which are otherwise not practicable (e.g., analysis of many organs from exposed animals, and human biomonitoring studies, and automation gives more uniform sample treatment and less dependence on operator performance. The high throughput modifications now available vary largely in their versatility, capacity, complexity and costs. The bottleneck for further increase of throughput appears to be the scoring.

  4. Modelling the predictive performance of credit scoring

    Directory of Open Access Journals (Sweden)

    Shi-Wei Shen

    2013-07-01

    Research purpose: The purpose of this empirical paper was to examine the predictive performance of credit scoring systems in Taiwan. Motivation for the study: Corporate lending remains a major business line for financial institutions. However, in light of the recent global financial crises, it has become extremely important for financial institutions to implement rigorous means of assessing clients seeking access to credit facilities. Research design, approach and method: Using a data sample of 10 349 observations drawn between 1992 and 2010, logistic regression models were utilised to examine the predictive performance of credit scoring systems. Main findings: A test of Goodness of fit demonstrated that credit scoring models that incorporated the Taiwan Corporate Credit Risk Index (TCRI, micro- and also macroeconomic variables possessed greater predictive power. This suggests that macroeconomic variables do have explanatory power for default credit risk. Practical/managerial implications: The originality in the study was that three models were developed to predict corporate firms’ defaults based on different microeconomic and macroeconomic factors such as the TCRI, asset growth rates, stock index and gross domestic product. Contribution/value-add: The study utilises different goodness of fits and receiver operator characteristics during the examination of the robustness of the predictive power of these factors.

  5. Resiliency scoring for business continuity plans.

    Science.gov (United States)

    Olson, Anna; Anderson, Jamie

    Through this paper readers will learn of a scoring methodology, referred to as resiliency scoring, which enables the evaluation of business continuity plans based upon analysis of their alignment with a predefined set of criteria that can be customised and are adaptable to the needs of any organisation. This patent pending tool has been successful in driving engagement and is a powerful resource to improve reporting capabilities, identify risks and gauge organisational resilience. The role of business continuity professionals is to aid their organisations in planning and preparedness activities aimed at mitigating the impacts of potential disruptions and ensuring critical business functions can continue in the event of unforeseen circumstances. This may seem like a daunting task for what can typically be a small team of individuals. For this reason, it is important to be able to leverage industry standards, documented best practices and effective tools to streamline and support your continuity programme. The resiliency scoring methodology developed and implemented at Target has proven to be a valuable tool in taking the organisation's continuity programme to the next level. This paper will detail how the tool was developed and provide guidance on how it can be customised to fit your organisation's unique needs.

  6. Soetomo score: score model in early identification of acute haemorrhagic stroke

    Directory of Open Access Journals (Sweden)

    Moh Hasan Machfoed

    2016-06-01

    Full Text Available Aim of the study: On financial or facility constraints of brain imaging, score model is used to predict the occurrence of acute haemorrhagic stroke. Accordingly, this study attempts to develop a new score model, called Soetomo score. Material and methods: The researchers performed a cross-sectional study of 176 acute stroke patients with onset of ≤24 hours who visited emergency unit of Dr. Soetomo Hospital from July 14th to December 14th, 2014. The diagnosis of haemorrhagic stroke was confirmed by head computed tomography scan. There were seven predictors of haemorrhagic stroke which were analysed by using bivariate and multivariate analyses. Furthermore, a multiple discriminant analysis resulted in an equation of Soetomo score model. The receiver operating characteristic procedure resulted in the values of area under curve and intersection point identifying haemorrhagic stroke. Afterward, the diagnostic test value was determined. Results: The equation of Soetomo score model was (3 × loss of consciousness + (3.5 × headache + (4 × vomiting − 4.5. Area under curve value of this score was 88.5% (95% confidence interval = 83.3–93.7%. In the Soetomo score model value of ≥−0.75, the score reached the sensitivity of 82.9%, specificity of 83%, positive predictive value of 78.8%, negative predictive value of 86.5%, positive likelihood ratio of 4.88, negative likelihood ratio of 0.21, false negative of 17.1%, false positive of 17%, and accuracy of 83%. Conclusions: The Soetomo score model value of ≥−0.75 can identify acute haemorrhagic stroke properly on the financial or facility constrains of brain imaging.

  7. Methods to score vertebral deformities in patients with rheumatoid arthritis

    NARCIS (Netherlands)

    Lems, W. F.; Jahangier, Z. N.; Raymakers, J. A.; Jacobs, J. W.; Bijlsma, J. W.

    1997-01-01

    The objective was to compare four different scoring methods for vertebral deformities: the semiquantitative Kleerekoper score and three quantitative scores (according to Minne, Melton and Raymakers) in patients with rheumatoid arthritis (RA). Lateral radiographs of the thoracic and lumbar vertebral

  8. siMS Score: Simple Method for Quantifying Metabolic Syndrome.

    Science.gov (United States)

    Soldatovic, Ivan; Vukovic, Rade; Culafic, Djordje; Gajic, Milan; Dimitrijevic-Sreckovic, Vesna

    2016-01-01

    To evaluate siMS score and siMS risk score, novel continuous metabolic syndrome scores as methods for quantification of metabolic status and risk. Developed siMS score was calculated using formula: siMS score = 2*Waist/Height + Gly/5.6 + Tg/1.7 + TAsystolic/130-HDL/1.02 or 1.28 (for male or female subjects, respectively). siMS risk score was calculated using formula: siMS risk score = siMS score * age/45 or 50 (for male or female subjects, respectively) * family history of cardio/cerebro-vascular events (event = 1.2, no event = 1). A sample of 528 obese and non-obese participants was used to validate siMS score and siMS risk score. Scores calculated as sum of z-scores (each component of metabolic syndrome regressed with age and gender) and sum of scores derived from principal component analysis (PCA) were used for evaluation of siMS score. Variants were made by replacing glucose with HOMA in calculations. Framingham score was used for evaluation of siMS risk score. Correlation between siMS score with sum of z-scores and weighted sum of factors of PCA was high (r = 0.866 and r = 0.822, respectively). Correlation between siMS risk score and log transformed Framingham score was medium to high for age groups 18+,30+ and 35+ (0.835, 0.707 and 0.667, respectively). siMS score and siMS risk score showed high correlation with more complex scores. Demonstrated accuracy together with superior simplicity and the ability to evaluate and follow-up individual patients makes siMS and siMS risk scores very convenient for use in clinical practice and research as well.

  9. Standardized UXO Demonstration Site Blind Grid Scoring Record No. 690

    National Research Council Canada - National Science Library

    Overbay, Larry, Jr; Archiable, Robert; McClung, Christina; Robitaille, George

    2005-01-01

    ...) utilizing the YPG Standardized UXO Technology Demonstration Site Blind Grid. The scoring record was coordinated by Larry Overbay and by the Standardized UXO Technology Demonstration Scoring Committee...

  10. Standardized UXO Technology Demonstration Site Blind Grid Scoring Record #833

    National Research Council Canada - National Science Library

    Fling, Rick; McClung, Christina; Burch, William; McDonnell, Patrick

    2007-01-01

    ...) utilizing the APG Standardized UXO Technology Demonstration Site Blind Grid. This Scoring Record was coordinated by Dennis Teefy and the Standardized UXO Technology Demonstration Site Scoring Committee...

  11. Standardized UXO Technology Demonstration Site, Woods Scoring Record Number 486

    National Research Council Canada - National Science Library

    Overbay, Larry; Robitaille, George

    2005-01-01

    ...) utilizing the APG Standardized UXO Technology Demonstration Site Open Field. The scoring record was coordinated by Larry Overbay and the Standardized UXO Technology Demonstration Site Scoring Committee...

  12. siMS Score: Simple Method for Quantifying Metabolic Syndrome

    OpenAIRE

    Soldatovic, Ivan; Vukovic, Rade; Culafic, Djordje; Gajic, Milan; Dimitrijevic-Sreckovic, Vesna

    2016-01-01

    Objective To evaluate siMS score and siMS risk score, novel continuous metabolic syndrome scores as methods for quantification of metabolic status and risk. Materials and Methods Developed siMS score was calculated using formula: siMS score = 2*Waist/Height + Gly/5.6 + Tg/1.7 + TAsystolic/130?HDL/1.02 or 1.28 (for male or female subjects, respectively). siMS risk score was calculated using formula: siMS risk score = siMS score * age/45 or 50 (for male or female subjects, respectively) * famil...

  13. Diagnostic accuracy of guys Hospital stroke score (allen score) in acute supratentorial thrombotic/haemorrhagic stroke

    International Nuclear Information System (INIS)

    Zulfiqar, A.; Toori, K. U.; Khan, S. S.; Hamza, M. I. M.; Zaman, S. U.

    2006-01-01

    A consecutive series of 103 patients, 58% male with mean age of 62 year (range 40-75 years), admitted with supratentorial stroke in our teaching hospital were studied. All patients had Computer Tomography scan brain done after clinical evaluation and application of Allen stroke score. Computer Tomography Scan confirmed thrombotic stroke in 55 (53%) patients and haemorrhagic stroke in 48 (47%) patients. Out of the 55 patients with definitive thrombotic stroke on Computer Tomography Scan, Allen stroke score suggested infarction in 67%, haemorrhage in 6% and remained inconclusive in 27% of cases. In 48 patients with definitive haemorrhagic stroke on Computer Tomography Scan, Allen stroke score suggested haemorrhage in 60%, infarction in 11% and remained inconclusive in 29% of cases. The overall accuracy of Allen stroke score was 66%. (author)

  14. Evaluation of modified Alvarado scoring system and RIPASA scoring system as diagnostic tools of acute appendicitis.

    Science.gov (United States)

    Shuaib, Abdullah; Shuaib, Ali; Fakhra, Zainab; Marafi, Bader; Alsharaf, Khalid; Behbehani, Abdullah

    2017-01-01

    Acute appendicitis is the most common surgical condition presented in emergency departments worldwide. Clinical scoring systems, such as the Alvarado and modified Alvarado scoring systems, were developed with the goal of reducing the negative appendectomy rate to 5%-10%. The Raja Isteri Pengiran Anak Saleha Appendicitis (RIPASA) scoring system was established in 2008 specifically for Asian populations. The aim of this study was to compare the modified Alvarado with the RIPASA scoring system in Kuwait population. This study included 180 patients who underwent appendectomies and were documented as having "acute appendicitis" or "abdominal pain" in the operating theatre logbook (unit B) from November 2014 to March 2016. The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), diagnostic accuracy, predicted negative appendectomy and receiver operating characteristic (ROC) curve of the modified Alvarado and RIPASA scoring systems were derived using SPSS statistical software. A total of 136 patients were included in this study according to our criteria. The cut-off threshold point of the modified Alvarado score was set at 7.0, which yielded a sensitivity of 82.8% and a specificity of 56%. The PPV was 89.3% and the NPV was 42.4%. The cut-off threshold point of the RIPASA score was set at 7.5, which yielded a 94.5% sensitivity and an 88% specificity. The PPV was 97.2% and the NPV was 78.5%. The predicted negative appendectomy rates were 10.7% and 2.2% for the modified Alvarado and RIPASA scoring systems, respectively. The negative appendectomy rate decreased significantly, from 18.4% to 10.7% for the modified Alvarado, and to 2.2% for the RIPASA scoring system, which was a significant difference (PAsian populations. It consists of 14 clinical parameters that can be obtained from a good patient history, clinical examination and laboratory investigations. The RIPASA scoring system is more accurate and specific than the modified Alvarado

  15. A Novel Scoring System Approach to Assess Patients with Lyme Disease (Nutech Functional Score)

    OpenAIRE

    Geeta Shroff; Petra Hopf-Seidel

    2018-01-01

    Introduction: A bacterial infection by Borrelia burgdorferi referred to as Lyme disease (LD) or borreliosis is transmitted mostly by a bite of the tick Ixodes scapularis in the USA and Ixodes ricinus in Europe. Various tests are used for the diagnosis of LD, but their results are often unreliable. We compiled a list of clinically visible and patient-reported symptoms that are associated with LD. Based on this list, we developed a novel scoring system. Methodology: Nutech functional Score (NF...

  16. External validation of the NOBLADS score, a risk scoring system for severe acute lower gastrointestinal bleeding.

    Directory of Open Access Journals (Sweden)

    Tomonori Aoki

    Full Text Available We aimed to evaluate the generalizability of NOBLADS, a severe lower gastrointestinal bleeding (LGIB prediction model which we had previously derived when working at a different institution, using an external validation cohort. NOBLADS comprises the following factors: non-steroidal anti-inflammatory drug use, no diarrhea, no abdominal tenderness, blood pressure ≤ 100 mmHg, antiplatelet drug use, albumin < 3.0 g/dL, disease score ≥ 2, and syncope.We retrospectively analyzed 511 patients emergently hospitalized for acute LGIB at the University of Tokyo Hospital, from January 2009 to August 2016. The areas under the receiver operating characteristic curves (ROCs-AUCs for severe bleeding (continuous and/or recurrent bleeding were compared between the original derivation cohort and the external validation cohort.Severe LGIB occurred in 44% of patients. Several clinical factors were significantly different between the external and derivation cohorts (p < 0.05, including background, laboratory data, NOBLADS scores, and diagnosis. The NOBLADS score predicted the severity of LGIB with an AUC value of 0.74 in the external validation cohort and one of 0.77 in the derivation cohort. In the external validation cohort, the score predicted the risk for blood transfusion need (AUC, 0.71, but was not adequate for predicting intervention need (AUC, 0.54. The in-hospital mortality rate was higher in patients with a score ≥ 5 than in those with a score < 5 (AUC, 0.83.Although the external validation cohort clinically differed from the derivation cohort in many ways, we confirmed the moderately high generalizability of NOBLADS, a clinical risk score for severe LGIB. Appropriate triage using this score may support early decision-making in various hospitals.

  17. The Veterans Affairs Cardiac Risk Score: Recalibrating the Atherosclerotic Cardiovascular Disease Score for Applied Use.

    Science.gov (United States)

    Sussman, Jeremy B; Wiitala, Wyndy L; Zawistowski, Matthew; Hofer, Timothy P; Bentley, Douglas; Hayward, Rodney A

    2017-09-01

    Accurately estimating cardiovascular risk is fundamental to good decision-making in cardiovascular disease (CVD) prevention, but risk scores developed in one population often perform poorly in dissimilar populations. We sought to examine whether a large integrated health system can use their electronic health data to better predict individual patients' risk of developing CVD. We created a cohort using all patients ages 45-80 who used Department of Veterans Affairs (VA) ambulatory care services in 2006 with no history of CVD, heart failure, or loop diuretics. Our outcome variable was new-onset CVD in 2007-2011. We then developed a series of recalibrated scores, including a fully refit "VA Risk Score-CVD (VARS-CVD)." We tested the different scores using standard measures of prediction quality. For the 1,512,092 patients in the study, the Atherosclerotic cardiovascular disease risk score had similar discrimination as the VARS-CVD (c-statistic of 0.66 in men and 0.73 in women), but the Atherosclerotic cardiovascular disease model had poor calibration, predicting 63% more events than observed. Calibration was excellent in the fully recalibrated VARS-CVD tool, but simpler techniques tested proved less reliable. We found that local electronic health record data can be used to estimate CVD better than an established risk score based on research populations. Recalibration improved estimates dramatically, and the type of recalibration was important. Such tools can also easily be integrated into health system's electronic health record and can be more readily updated.

  18. Pediatric siMS score: A new, simple and accurate continuous metabolic syndrome score for everyday use in pediatrics.

    Science.gov (United States)

    Vukovic, Rade; Milenkovic, Tatjana; Stojan, George; Vukovic, Ana; Mitrovic, Katarina; Todorovic, Sladjana; Soldatovic, Ivan

    2017-01-01

    The dichotomous nature of the current definition of metabolic syndrome (MS) in youth results in loss of information. On the other hand, the calculation of continuous MS scores using standardized residuals in linear regression (Z scores) or factor scores of principal component analysis (PCA) is highly impractical for clinical use. Recently, a novel, easily calculated continuous MS score called siMS score was developed based on the IDF MS criteria for the adult population. To develop a Pediatric siMS score (PsiMS), a modified continuous MS score for use in the obese youth, based on the original siMS score, while keeping the score as simple as possible and retaining high correlation with more complex scores. The database consisted of clinical data on 153 obese (BMI ≥95th percentile) children and adolescents. Continuous MS scores were calculated using Z scores and PCA, as well as the original siMS score. Four variants of PsiMS score were developed in accordance with IDF criteria for MS in youth and correlation of these scores with PCA and Z score derived MS continuous scores was assessed. PsiMS score calculated using formula: (2xWaist/Height) + (Glucose(mmol/l)/5.6) + (triglycerides(mmol/l)/1.7) + (Systolic BP/130)-(HDL(mmol/l)/1.02) showed the highest correlation with most of the complex continuous scores (0.792-0.901). The original siMS score also showed high correlation with continuous MS scores. PsiMS score represents a practical and accurate score for the evaluation of MS in the obese youth. The original siMS score should be used when evaluating large cohorts consisting of both adults and children.

  19. Scoring function to predict solubility mutagenesis

    Directory of Open Access Journals (Sweden)

    Deutsch Christopher

    2010-10-01

    Full Text Available Abstract Background Mutagenesis is commonly used to engineer proteins with desirable properties not present in the wild type (WT protein, such as increased or decreased stability, reactivity, or solubility. Experimentalists often have to choose a small subset of mutations from a large number of candidates to obtain the desired change, and computational techniques are invaluable to make the choices. While several such methods have been proposed to predict stability and reactivity mutagenesis, solubility has not received much attention. Results We use concepts from computational geometry to define a three body scoring function that predicts the change in protein solubility due to mutations. The scoring function captures both sequence and structure information. By exploring the literature, we have assembled a substantial database of 137 single- and multiple-point solubility mutations. Our database is the largest such collection with structural information known so far. We optimize the scoring function using linear programming (LP methods to derive its weights based on training. Starting with default values of 1, we find weights in the range [0,2] so that predictions of increase or decrease in solubility are optimized. We compare the LP method to the standard machine learning techniques of support vector machines (SVM and the Lasso. Using statistics for leave-one-out (LOO, 10-fold, and 3-fold cross validations (CV for training and prediction, we demonstrate that the LP method performs the best overall. For the LOOCV, the LP method has an overall accuracy of 81%. Availability Executables of programs, tables of weights, and datasets of mutants are available from the following web page: http://www.wsu.edu/~kbala/OptSolMut.html.

  20. Best waveform score for diagnosing keratoconus

    Directory of Open Access Journals (Sweden)

    Allan Luz

    2013-12-01

    Full Text Available PURPOSE: To test whether corneal hysteresis (CH and corneal resistance factor (CRF can discriminate between keratoconus and normal eyes and to evaluate whether the averages of two consecutive measurements perform differently from the one with the best waveform score (WS for diagnosing keratoconus. METHODS: ORA measurements for one eye per individual were selected randomly from 53 normal patients and from 27 patients with keratoconus. Two groups were considered the average (CH-Avg, CRF-Avg and best waveform score (CH-WS, CRF-WS groups. The Mann-Whitney U-test was used to evaluate whether the variables had similar distributions in the Normal and Keratoconus groups. Receiver operating characteristics (ROC curves were calculated for each parameter to assess the efficacy for diagnosing keratoconus and the same obtained for each variable were compared pairwise using the Hanley-McNeil test. RESULTS: The CH-Avg, CRF-Avg, CH-WS and CRF-WS differed significantly between the normal and keratoconus groups (p<0.001. The areas under the ROC curve (AUROC for CH-Avg, CRF-Avg, CH-WS, and CRF-WS were 0.824, 0.873, 0.891, and 0.931, respectively. CH-WS and CRF-WS had significantly better AUROCs than CH-Avg and CRF-Avg, respectively (p=0.001 and 0.002. CONCLUSION: The analysis of the biomechanical properties of the cornea through the ORA method has proved to be an important aid in the diagnosis of keratoconus, regardless of the method used. The best waveform score (WS measurements were superior to the average of consecutive ORA measurements for diagnosing keratoconus.

  1. Setting pass scores for clinical skills assessment.

    Science.gov (United States)

    Liu, Min; Liu, Keh-Min

    2008-12-01

    In a clinical skills assessment, the decision to pass or fail an examinee should be based on the test content or on the examinees' performance. The process of deciding a pass score is known as setting a standard of the examination. This requires a properly selected panel of expert judges and a suitable standard setting method, which best fits the purpose of the examination. Six standard setting methods that are often used in clinical skills assessment are described to provide an overview of the standard setting process.

  2. Setting Pass Scores for Clinical Skills Assessment

    Directory of Open Access Journals (Sweden)

    Min Liu

    2008-12-01

    Full Text Available In a clinical skills assessment, the decision to pass or fail an examinee should be based on the test content or on the examinees' performance. The process of deciding a pass score is known as setting a standard of the examination. This requires a properly selected panel of expert judges and a suitable standard setting method, which best fits the purpose of the examination. Six standard setting methods that are often used in clinical skills assessment are described to provide an overview of the standard setting process.

  3. Sway Area and Velocity Correlated With MobileMat Balance Error Scoring System (BESS) Scores.

    Science.gov (United States)

    Caccese, Jaclyn B; Buckley, Thomas A; Kaminski, Thomas W

    2016-08-01

    The Balance Error Scoring System (BESS) is often used for sport-related concussion balance assessment. However, moderate intratester and intertester reliability may cause low initial sensitivity, suggesting that a more objective balance assessment method is needed. The MobileMat BESS was designed for objective BESS scoring, but the outcome measures must be validated with reliable balance measures. Thus, the purpose of this investigation was to compare MobileMat BESS scores to linear and nonlinear measures of balance. Eighty-eight healthy collegiate student-athletes (age: 20.0 ± 1.4 y, height: 177.7 ± 10.7 cm, mass: 74.8 ± 13.7 kg) completed the MobileMat BESS. MobileMat BESS scores were compared with 95% area, sway velocity, approximate entropy, and sample entropy. MobileMat BESS scores were significantly correlated with 95% area for single-leg (r = .332) and tandem firm (r = .474), and double-leg foam (r = .660); and with sway velocity for single-leg (r = .406) and tandem firm (r = .601), and double-leg (r = .575) and single-leg foam (r = .434). MobileMat BESS scores were not correlated with approximate or sample entropy. MobileMat BESS scores were low to moderately correlated with linear measures, suggesting the ability to identify changes in the center of mass-center of pressure relationship, but not higher-order processing associated with nonlinear measures. These results suggest that the MobileMat BESS may be a clinically-useful tool that provides objective linear balance measures.

  4. Ripasa score: a new diagnostic score for diagnosis of acute appendicitis

    International Nuclear Information System (INIS)

    Butt, M.Q.

    2014-01-01

    Objective: To determine the usefulness of RIPASA score for the diagnosis of acute appendicitis using histopathology as a gold standard. Study Design: Cross-sectional study. Place and Duration of Study: Department of General Surgery, Combined Military Hospital, Kohat, from September 2011 to March 2012. Methodology: A total of 267 patients were included in this study. RIPASA score was assessed. The diagnosis of appendicitis was made clinically aided by routine sonography of abdomen. After appendicectomies, resected appendices were sent for histopathological examination. The 15 parameters and the scores generated were age (less than 40 years = 1 point; greater than 40 years = 0.5 point), gender (male = 1 point; female = 0.5 point), Right Iliac Fossa (RIF) pain (0.5 point), migration of pain to RIF (0.5 point), nausea and vomiting (1 point), anorexia (1 point), duration of symptoms (less than 48 hours = 1 point; more than 48 hours = 0.5 point), RIF tenderness (1 point), guarding (2 points), rebound tenderness (1 point), Rovsing's sign (2 points), fever (1 point), raised white cell count (1 point), negative urinalysis (1 point) and foreign national registration identity card (1 point). The optimal cut-off threshold score from the ROC was 7.5. Sensitivity analysis was done. Results: Out of 267 patients, 156 (58.4%) were male while remaining 111 patients (41.6%) were female with mean age of 23.5 +- 9.1 years. Sensitivity of RIPASA score was 96.7%, specificity 93.0%, diagnostic accuracy was 95.1%, positive predictive value was 94.8% and negative predictive value was 95.54%. Conclusion: RIPASA score at a cut-off total score of 7.5 was a useful tool to diagnose appendicitis, in equivocal cases of pain. (author)

  5. How is the injury severity scored? a brief review of scoring systems

    Directory of Open Access Journals (Sweden)

    Mohsen Ebrahimi

    2015-06-01

    Full Text Available The management of injured patients is a critical issue in pre-hospital and emergency departments. Trauma victims are usually young and the injuries may lead to mortality or severe morbidities. The severity of injury can be estimated by observing the anatomic and physiologic evidences. Scoring systems are used to present a scale of describing the severity of the injuries in the victims.We reviewed the evidences of famous scoring systems, the history of their development, applications and their evolutions. We searched electronic database PubMed and Google scholar with keywords: (trauma OR injury AND (severity OR intensity AND (score OR scale.In this paper, we are going to present a definition of scoring systems and discuss the Abbreviated Injury Scale (AIS and Injury Severity Score (ISS, the most acceptable systems, their applications and their advantages and limitations.Several injury-scoring methods have been introduced. Each method has specific features, advantages and disadvantages. The AIS is an anatomical-based scoring system, which provides a standard numerical scale of ranking and comparing injuries. The ISS was established as a platform for trauma data registry. ISS is also an anatomically-based ordinal scale, with a range of 1-75. Several databases and studies are formed based on ISS and are available for trauma management research.Although the ISS is not perfect, it is established as the basic platform of health services and public health researches. The ISS registering system can provide many opportunities for the development of efficient data recording and statistical analyzing models.

  6. High-Throughput Scoring of Seed Germination.

    Science.gov (United States)

    Ligterink, Wilco; Hilhorst, Henk W M

    2017-01-01

    High-throughput analysis of seed germination for phenotyping large genetic populations or mutant collections is very labor intensive and would highly benefit from an automated setup. Although very often used, the total germination percentage after a nominated period of time is not very informative as it lacks information about start, rate, and uniformity of germination, which are highly indicative of such traits as dormancy, stress tolerance, and seed longevity. The calculation of cumulative germination curves requires information about germination percentage at various time points. We developed the GERMINATOR package: a simple, highly cost-efficient, and flexible procedure for high-throughput automatic scoring and evaluation of germination that can be implemented without the use of complex robotics. The GERMINATOR package contains three modules: (I) design of experimental setup with various options to replicate and randomize samples; (II) automatic scoring of germination based on the color contrast between the protruding radicle and seed coat on a single image; and (III) curve fitting of cumulative germination data and the extraction, recap, and visualization of the various germination parameters. GERMINATOR is a freely available package that allows the monitoring and analysis of several thousands of germination tests, several times a day by a single person.

  7. Development of a severity score for CRPS.

    Science.gov (United States)

    Harden, R Norman; Bruehl, Stephen; Perez, Roberto S G M; Birklein, Frank; Marinus, Johan; Maihofner, Christian; Lubenow, Timothy; Buvanendran, Asokumar; Mackey, Sean; Graciosa, Joseph; Mogilevski, Mila; Ramsden, Christopher; Schlereth, Tanja; Chont, Melissa; Vatine, Jean-Jacques

    2010-12-01

    The clinical diagnosis of Complex Regional Pain Syndrome (CRPS) is a dichotomous (yes/no) categorization necessary for clinical decision-making. However, such dichotomous diagnostic categories do not convey an individual's subtle and temporal gradations in severity of the condition, and have poor statistical power when used as an outcome measure in research. This study evaluated the validity and potential utility of a continuous type score to index severity of CRPS. Psychometric and medical evaluations were conducted in 114 CRPS patients and 41 non-CRPS neuropathic pain patients. Based on the presence/absence of 17 clinically-assessed signs and symptoms of CRPS, an overall CRPS Severity Score (CSS) was derived. The CSS discriminated well between CRPS and non-CRPS patients (pCRPS diagnoses using both IASP diagnostic criteria (Eta=0.69) and proposed revised criteria (Eta=0.77-0.88). Higher CSS was associated with significantly higher clinical pain intensity, distress, and functional impairments, as well as greater bilateral temperature asymmetry and thermal perception abnormalities (p'sCRPS, and support its validity as an index of CRPS severity. Its utility as an outcome measure in research studies is also suggested, with potential statistical advantages over dichotomous diagnostic criteria. Copyright © 2010. Published by Elsevier B.V.

  8. Knee Injury and Osteoarthritis Outcome Score (KOOS)

    DEFF Research Database (Denmark)

    Collins, N J; Prinsen, C A C; Christensen, R

    2016-01-01

    OBJECTIVE: To conduct a systematic review and meta-analysis to synthesize evidence regarding measurement properties of the Knee injury and Osteoarthritis Outcome Score (KOOS). DESIGN: A comprehensive literature search identified 37 eligible papers evaluating KOOS measurement properties in partici......OBJECTIVE: To conduct a systematic review and meta-analysis to synthesize evidence regarding measurement properties of the Knee injury and Osteoarthritis Outcome Score (KOOS). DESIGN: A comprehensive literature search identified 37 eligible papers evaluating KOOS measurement properties...... in participants with knee injuries and/or osteoarthritis (OA). Methodological quality was evaluated using the COSMIN checklist. Where possible, meta-analysis of extracted data was conducted for all studies and stratified by age and knee condition; otherwise narrative synthesis was performed. RESULTS: KOOS has...... adequate internal consistency, test-retest reliability and construct validity in young and old adults with knee injuries and/or OA. The ADL subscale has better content validity for older patients and Sport/Rec for younger patients with knee injuries, while the Pain subscale is more relevant for painful...

  9. Literature in focus: How to Score

    CERN Document Server

    2006-01-01

    What is the perfect way to take a free kick? Which players are under more stress: attackers, midfielders or defenders? How do we know when a ball has crossed the goal-line? And how can teams win a penalty shoot out? From international team formations to the psychology of the pitch and the changing room... The World Cup might be a time to forget about physics for a while, but not for Ken Bray, a theoretical physicist and visiting Fellow of the Sport and Exercise Science Group at the University of Bath who specializes in the science of football. Dr Bray will visit CERN to talk exclusively about his book: How to Score. As a well-seasoned speaker and advisor to professional football teams, this presentation promises to be a fascinating and timely insight into the secret science that lies behind 'the beautiful game'. If you play or just watch football, don't miss this event! Ken Bray - How to Score Thursday 22 June at 3 p.m. (earlier than usual to avoid clashes with World Cup matches!) Central Library reading ...

  10. 24 CFR 902.45 - Management operations scoring and thresholds.

    Science.gov (United States)

    2010-04-01

    ... 24 Housing and Urban Development 4 2010-04-01 2010-04-01 false Management operations scoring and... URBAN DEVELOPMENT PUBLIC HOUSING ASSESSMENT SYSTEM PHAS Indicator #3: Management Operations § 902.45 Management operations scoring and thresholds. (a) Scoring. The Management Operations Indicator score provides...

  11. Conditional Standard Errors of Measurement for Scale Scores.

    Science.gov (United States)

    Kolen, Michael J.; And Others

    1992-01-01

    A procedure is described for estimating the reliability and conditional standard errors of measurement of scale scores incorporating the discrete transformation of raw scores to scale scores. The method is illustrated using a strong true score model, and practical applications are described. (SLD)

  12. Validating the Interpretations and Uses of Test Scores

    Science.gov (United States)

    Kane, Michael T.

    2013-01-01

    To validate an interpretation or use of test scores is to evaluate the plausibility of the claims based on the scores. An argument-based approach to validation suggests that the claims based on the test scores be outlined as an argument that specifies the inferences and supporting assumptions needed to get from test responses to score-based…

  13. [The diagnostic and the exclusion scores for pulmonary embolism].

    Science.gov (United States)

    Junod, A

    2015-05-27

    Several clinical scores for the diagnosis of pulmonary embolism (PE) have been published. The most popular ones are the Wells score and the revised Geneva score; simplified versions exist for these two scores; they have been validated. Both scores have common properties, but there is a major difference for the Wells score, namely the inclusion of a feature based on clinical judgment. These two scores in combination with D-dimers measurement have been used to rule out PE. An important improvement in this process has recently taken place with the use of an adjustable, age-dependent threshold for DD for patients over 50 years.

  14. Nursing Activities Score and Acute Kidney Injury.

    Science.gov (United States)

    Coelho, Filipe Utuari de Andrade; Watanabe, Mirian; Fonseca, Cassiane Dezoti da; Padilha, Katia Grillo; Vattimo, Maria de Fátima Fernandes

    2017-01-01

    to evaluate the nursing workload in intensive care patients with acute kidney injury (AKI). A quantitative study, conducted in an intensive care unit, from April to August of 2015. The Nursing Activities Score (NAS) and Kidney Disease Improving Global Outcomes (KDIGO) were used to measure nursing workload and to classify the stage of AKI, respectively. A total of 190 patients were included. Patients who developed AKI (44.2%) had higher NAS when compared to those without AKI (43.7% vs 40.7%), p <0.001. Patients with stage 1, 2 and 3 AKI showed higher NAS than those without AKI. A relationship was identified between stage 2 and 3 with those without AKI (p = 0.002 and p <0.001). The NAS was associated with the presence of AKI, the score increased with the progression of the stages, and it was associated with AKI, stage 2 and 3. avaliar a carga de trabalho de enfermagem em pacientes de terapia intensiva com lesão renal aguda (LRA). estudo quantitativo, em Unidade de Terapia Intensiva, no período de abril a agosto de 2015. O Nursing Activities Score (NAS) e o Kidney Disease Improving Global Outcomes (KDIGO) foram utilizados para medir a carga de trabalho de enfermagem e classificar o estágio da LRA, respectivamente. foram incluídos 190 pacientes. Os pacientes que desenvolveram LRA (44,2%) possuíam NAS superiores quando comparados aos sem LRA (43,7% vs 40,7%), p<0,001. Os pacientes com LRA nos estágios 1, 2 e 3 de LRA demonstraram NAS superiores aos sem LRA, houve relação entre os estágios 2 e 3 com os sem LRA, p=0,002 e p<0,001. o NAS apresentou associação com a existência de LRA, visto que seu valor aumenta com a progressão dos estágios, tendo associação com os estágios 2 e 3 de LRA.

  15. Reproducibility of scoring emphysema by HRCT

    International Nuclear Information System (INIS)

    Malinen, A.; Partanen, K.; Rytkoenen, H.; Vanninen, R.; Erkinjuntti-Pekkanen, R.

    2002-01-01

    Purpose: We evaluated the reproducibility of three visual scoring methods of emphysema and compared these methods with pulmonary function tests (VC, DLCO, FEV1 and FEV%) among farmer's lung patients and farmers. Material and Methods: Three radiologists examined high-resolution CT images of farmer's lung patients and their matched controls (n=70) for chronic interstitial lung diseases. Intraobserver reproducibility and interobserver variability were assessed for three methods: severity, Sanders' (extent) and Sakai. Pulmonary function tests as spirometry and diffusing capacity were measured. Results: Intraobserver -values for all three methods were good (0.51-0.74). Interobserver varied from 0.35 to 0.72. The Sanders' and the severity methods correlated strongly with pulmonary function tests, especially DLCO and FEV1. Conclusion: The Sanders' method proved to be reliable in evaluating emphysema, in terms of good consistency of interpretation and good correlation with pulmonary function tests

  16. Reproducibility of scoring emphysema by HRCT

    Energy Technology Data Exchange (ETDEWEB)

    Malinen, A.; Partanen, K.; Rytkoenen, H.; Vanninen, R. [Kuopio Univ. Hospital (Finland). Dept. of Clinical Radiology; Erkinjuntti-Pekkanen, R. [Kuopio Univ. Hospital (Finland). Dept. of Pulmonary Diseases

    2002-04-01

    Purpose: We evaluated the reproducibility of three visual scoring methods of emphysema and compared these methods with pulmonary function tests (VC, DLCO, FEV1 and FEV%) among farmer's lung patients and farmers. Material and Methods: Three radiologists examined high-resolution CT images of farmer's lung patients and their matched controls (n=70) for chronic interstitial lung diseases. Intraobserver reproducibility and interobserver variability were assessed for three methods: severity, Sanders' (extent) and Sakai. Pulmonary function tests as spirometry and diffusing capacity were measured. Results: Intraobserver -values for all three methods were good (0.51-0.74). Interobserver varied from 0.35 to 0.72. The Sanders' and the severity methods correlated strongly with pulmonary function tests, especially DLCO and FEV1. Conclusion: The Sanders' method proved to be reliable in evaluating emphysema, in terms of good consistency of interpretation and good correlation with pulmonary function tests.

  17. The Rectal Cancer Female Sexuality Score

    DEFF Research Database (Denmark)

    Thyø, Anne; Emmertsen, Katrine J; Laurberg, Søren

    2018-01-01

    BACKGROUND: Sexual dysfunction and impaired quality of life is a potential side effect to rectal cancer treatment. OBJECTIVE: The objective of this study was to develop and validate a simple scoring system intended to evaluate sexual function in women treated for rectal cancer. DESIGN......: This is a population-based cross-sectional study. SETTINGS: Female patients diagnosed with rectal cancer between 2001 and 2014 were identified by using the Danish Colorectal Cancer Group's database. Participants filled in the validated Sexual Function Vaginal Changes questionnaire. Women declared to be sexually active...... in the validation group. PATIENTS: Female patients with rectal cancer above the age of 18 who underwent abdominoperineal resection, Hartmann procedure, or total/partial mesorectal excision were selected. MAIN OUTCOME MEASURES: The primary outcome measured was the quality of life that was negatively affected because...

  18. ABOUT PSYCHOLOGICAL VARIABLES IN APPLICATION SCORING MODELS

    Directory of Open Access Journals (Sweden)

    Pablo Rogers

    2015-01-01

    Full Text Available The purpose of this study is to investigate the contribution of psychological variables and scales suggested by Economic Psychology in predicting individuals’ default. Therefore, a sample of 555 individuals completed a self-completion questionnaire, which was composed of psychological variables and scales. By adopting the methodology of the logistic regression, the following psychological and behavioral characteristics were found associated with the group of individuals in default: a negative dimensions related to money (suffering, inequality and conflict; b high scores on the self-efficacy scale, probably indicating a greater degree of optimism and over-confidence; c buyers classified as compulsive; d individuals who consider it necessary to give gifts to children and friends on special dates, even though many people consider this a luxury; e problems of self-control identified by individuals who drink an average of more than four glasses of alcoholic beverage a day.

  19. Do efficiency scores depend on input mix?

    DEFF Research Database (Denmark)

    Asmild, Mette; Hougaard, Jens Leth; Kronborg, Dorte

    2013-01-01

    In this paper we examine the possibility of using the standard Kruskal-Wallis (KW) rank test in order to evaluate whether the distribution of efficiency scores resulting from Data Envelopment Analysis (DEA) is independent of the input (or output) mix of the observations. Since the DEA frontier...... is estimated, many standard assumptions for evaluating the KW test statistic are violated. Therefore, we propose to explore its statistical properties by the use of simulation studies. The simulations are performed conditional on the observed input mixes. The method, unlike existing approaches...... the assumption of mix independence is rejected the implication is that it, for example, is impossible to determine whether machine intensive project are more or less efficient than labor intensive projects....

  20. 'Mechanical restraint-confounders, risk, alliance score'

    DEFF Research Database (Denmark)

    Deichmann Nielsen, Lea; Bech, Per; Hounsgaard, Lise

    2017-01-01

    . AIM: To clinically validate a new, structured short-term risk assessment instrument called the Mechanical Restraint-Confounders, Risk, Alliance Score (MR-CRAS), with the intended purpose of supporting the clinicians' observation and assessment of the patient's readiness to be released from mechanical...... restraint. METHODS: The content and layout of MR-CRAS and its user manual were evaluated using face validation by forensic mental health clinicians, content validation by an expert panel, and pilot testing within two, closed forensic mental health inpatient units. RESULTS: The three sub-scales (Confounders......, Risk, and a parameter of Alliance) showed excellent content validity. The clinical validations also showed that MR-CRAS was perceived and experienced as a comprehensible, relevant, comprehensive, and useable risk assessment instrument. CONCLUSIONS: MR-CRAS contains 18 clinically valid items...

  1. SOS score: an optimized score to screen acute stroke patients for obstructive sleep apnea.

    Science.gov (United States)

    Camilo, Millene R; Sander, Heidi H; Eckeli, Alan L; Fernandes, Regina M F; Dos Santos-Pontelli, Taiza E G; Leite, Joao P; Pontes-Neto, Octavio M

    2014-09-01

    Obstructive sleep apnea (OSA) is frequent in acute stroke patients, and has been associated with higher mortality and worse prognosis. Polysomnography (PSG) is the gold standard diagnostic method for OSA, but it is impracticable as a routine for all acute stroke patients. We evaluated the accuracy of two OSA screening tools, the Berlin Questionnaire (BQ), and the Epworth Sleepiness Scale (ESS) when administered to relatives of acute stroke patients; we also compared these tools against a combined screening score (SOS score). Ischemic stroke patients were submitted to a full PSG at the first night after onset of symptoms. OSA severity was measured by apnea-hypopnea index (AHI). BQ and ESS were administered to relatives of stroke patients before the PSG and compared to SOS score for accuracy and C-statistics. We prospectively studied 39 patients. OSA (AHI ≥10/h) was present in 76.9%. The SOS score [area under the curve (AUC): 0.812; P = 0.005] and ESS (AUC: 0.789; P = 0.009) had good predictive value for OSA. The SOS score was the only tool with significant predictive value (AUC: 0.686; P = 0.048) for severe OSA (AHI ≥30/h), when compared to ESS (P = 0.119) and BQ (P = 0.191). The threshold of SOS ≤10 showed high sensitivity (90%) and negative predictive value (96.2%) for OSA; SOS ≥20 showed high specificity (100%) and positive predictive value (92.5%) for severe OSA. The SOS score administered to relatives of stroke patients is a useful tool to screen for OSA and may decrease the need for PSG in acute stroke setting. Copyright © 2014 Elsevier B.V. All rights reserved.

  2. Prediction of true test scores from observed item scores and ancillary data.

    Science.gov (United States)

    Haberman, Shelby J; Yao, Lili; Sinharay, Sandip

    2015-05-01

    In many educational tests which involve constructed responses, a traditional test score is obtained by adding together item scores obtained through holistic scoring by trained human raters. For example, this practice was used until 2008 in the case of GRE(®) General Analytical Writing and until 2009 in the case of TOEFL(®) iBT Writing. With use of natural language processing, it is possible to obtain additional information concerning item responses from computer programs such as e-rater(®). In addition, available information relevant to examinee performance may include scores on related tests. We suggest application of standard results from classical test theory to the available data to obtain best linear predictors of true traditional test scores. In performing such analysis, we require estimation of variances and covariances of measurement errors, a task which can be quite difficult in the case of tests with limited numbers of items and with multiple measurements per item. As a consequence, a new estimation method is suggested based on samples of examinees who have taken an assessment more than once. Such samples are typically not random samples of the general population of examinees, so that we apply statistical adjustment methods to obtain the needed estimated variances and covariances of measurement errors. To examine practical implications of the suggested methods of analysis, applications are made to GRE General Analytical Writing and TOEFL iBT Writing. Results obtained indicate that substantial improvements are possible both in terms of reliability of scoring and in terms of assessment reliability. © 2015 The British Psychological Society.

  3. Coronary artery calcium scoring in myocardial infarction

    International Nuclear Information System (INIS)

    Beslic, S.; Dalagija, F.

    2005-01-01

    Background. The aim of this study was to evaluate coronary artery calcium scoring and the assessment of the risk factors in patients with myocardial infarction (MI). Methods. During the period of three years, 27 patients with MI were analyzed. The average age of patients was 66.1 years (46 to 81). Coronary arteries calcium was evaluated by multi row detector computed tomography (MTDC) S omatom Volume Zoom Siemens , and, retrospectively by ECG gating data acquisition. Semi automated calcium quantification to calculate Agatston calcium score (CS) was performed with 4 x 2.5 mm collimation, using 130 ml of contrast medium, injected with an automatic injector, with the flow rate of 4 ml/sec. The delay time was determined empirically. At the same time several risk factors were evaluated. Results. Out of 27 patients with MI, 3 (11.1%) patients had low CS (10- 100), 5 (18.5%) moderate CS (101- 499), and 19 (70.4%) patients high CS (>500). Of risk factors, smoking was confirmed in 17 (63.0%), high blood pressure (HTA) in 10 (57.0%), diabetes mellitus in 7 (25.9%), positive family history in 5 (18.5%), pathological lipids in 5 (18.5%), alcohol abuse in 4 (1.8%) patients. Six (22.2%) patients had symptoms of angina pectoris. Conclusions. The research showed high correlation of MI and high CS (>500). Smoking, HTA, diabetes mellitus, positive family history and hypercholesterolemia are significant risk factors. Symptoms are relatively poor in large number of patients. (author)

  4. Prediction of IOI-HA Scores Using Speech Reception Thresholds and Speech Discrimination Scores in Quiet

    DEFF Research Database (Denmark)

    Brännström, K Jonas; Lantz, Johannes; Nielsen, Lars Holme

    2014-01-01

    ), and speech discrimination scores (SDSs) in quiet or in noise are common assessments made prior to hearing aid (HA) fittings. It is not known whether SRT and SDS in quiet relate to HA outcome measured with the International Outcome Inventory for Hearing Aids (IOI-HA). PURPOSE: The aim of the present study...... COLLECTION AND ANALYSIS: The psychometric properties were evaluated and compared to previous studies using the IOI-HA. The associations and differences between the outcome scores and a number of descriptive variables (age, gender, fitted monaurally/binaurally with HA, first-time/experienced HA users, years...

  5. A Summary Score for the Framingham Heart Study Neuropsychological Battery.

    Science.gov (United States)

    Downer, Brian; Fardo, David W; Schmitt, Frederick A

    2015-10-01

    To calculate three summary scores of the Framingham Heart Study neuropsychological battery and determine which score best differentiates between subjects classified as having normal cognition, test-based impaired learning and memory, test-based multidomain impairment, and dementia. The final sample included 2,503 participants. Three summary scores were assessed: (a) composite score that provided equal weight to each subtest, (b) composite score that provided equal weight to each cognitive domain assessed by the neuropsychological battery, and (c) abbreviated score comprised of subtests for learning and memory. Receiver operating characteristic analysis was used to determine which summary score best differentiated between the four cognitive states. The summary score that provided equal weight to each subtest best differentiated between the four cognitive states. A summary score that provides equal weight to each subtest is an efficient way to utilize all of the cognitive data collected by a neuropsychological battery. © The Author(s) 2015.

  6. ITC Guidelines on Quality Control in Scoring, Test Analysis, and Reporting of Test Scores

    Science.gov (United States)

    Allalouf, Avi

    2014-01-01

    The Quality Control (QC) Guidelines are intended to increase the efficiency, precision, and accuracy of the scoring, analysis, and reporting process of testing. The QC Guidelines focus on large-scale testing operations where multiple forms of tests are created for use on set dates. However, they may also be used for a wide variety of other testing…

  7. Renal dysfunction in liver cirrhosis and its correlation with Child-Pugh score and MELD score

    Science.gov (United States)

    Siregar, G. A.; Gurning, M.

    2018-03-01

    Renal dysfunction (RD) is a serious and common complication in a patient with liver cirrhosis. It provides a poor prognosis. The aim of our study was to evaluate the renal function in liver cirrhosis, also to determine the correlation with the graduation of liver disease assessed by Child-Pugh Score (CPS) and MELD score. This was a cross-sectional study included patients with liver cirrhosis admitted to Adam Malik Hospital Medan in June - August 2016. We divided them into two groups as not having renal dysfunction (serum creatinine SPSS 22.0 was used. Statistical methods used: Chi-square, Fisher exact, one way ANOVA, Kruskal Wallis test and Pearson coefficient of correlation. The level of significance was p<0.05. 55 patients with presented with renal dysfunction were 16 (29.1 %). There was statistically significant inverse correlation between GFR and CPS (r = -0.308), GFR and MELD score (r = -0.278). There was a statistically significant correlation between creatinine and MELD score (r = 0.359), creatinine and CPS (r = 0.382). The increase of the degree of liver damage is related to the increase of renal dysfunction.

  8. A Novel Scoring System Approach to Assess Patients with Lyme Disease (Nutech Functional Score).

    Science.gov (United States)

    Shroff, Geeta; Hopf-Seidel, Petra

    2018-01-01

    A bacterial infection by Borrelia burgdorferi referred to as Lyme disease (LD) or borreliosis is transmitted mostly by a bite of the tick Ixodes scapularis in the USA and Ixodes ricinus in Europe. Various tests are used for the diagnosis of LD, but their results are often unreliable. We compiled a list of clinically visible and patient-reported symptoms that are associated with LD. Based on this list, we developed a novel scoring system. Nutech functional Score (NFS), which is a 43 point positional (every symptom is subgraded and each alternative gets some points according to its position) and directional (moves in direction bad to good) scoring system that assesses the patient's condition. The grades of the scoring system have been converted into numeric values for conducting probability based studies. Each symptom is graded from 1 to 5 that runs in direction BAD → GOOD. NFS is a unique tool that can be used universally to assess the condition of patients with LD.

  9. A Novel Scoring System Approach to Assess Patients with Lyme Disease (Nutech Functional Score

    Directory of Open Access Journals (Sweden)

    Geeta Shroff

    2018-01-01

    Full Text Available Introduction: A bacterial infection by Borrelia burgdorferi referred to as Lyme disease (LD or borreliosis is transmitted mostly by a bite of the tick Ixodes scapularis in the USA and Ixodes ricinus in Europe. Various tests are used for the diagnosis of LD, but their results are often unreliable. We compiled a list of clinically visible and patient-reported symptoms that are associated with LD. Based on this list, we developed a novel scoring system. Methodology: Nutech functional Score (NFS, which is a 43 point positional (every symptom is subgraded and each alternative gets some points according to its position and directional (moves in direction bad to good scoring system that assesses the patient's condition. Results: The grades of the scoring system have been converted into numeric values for conducting probability based studies. Each symptom is graded from 1 to 5 that runs in direction BAD → GOOD. Conclusion: NFS is a unique tool that can be used universally to assess the condition of patients with LD.

  10. Evaluation of classifiers that score linear type traits and body condition score using common sires

    NARCIS (Netherlands)

    Veerkamp, R.F.; Gerritsen, C.L.M.; Koenen, E.P.C.; Hamoen, A.; Jong, de G.

    2002-01-01

    Subjective visual assessment of animals by classifiers is undertaken for several different traits in farm livestock, e.g., linear type traits, body condition score, or carcass conformation. One of the difficulties in assessment is the effect of an individual classifier. To ensure that classifiers

  11. Symptom scoring systems to diagnose distal polyneuropathy in diabetes : the Diabetic Neuropathy Symptom score

    NARCIS (Netherlands)

    Meijer, J.W.G.; Smit, A.J.; van Sonderen, E.; Groothoff, J.W.; Eisma, W.H.; Links, T.P.

    2002-01-01

    AIMS: To provide one of the diagnostic categories for distal diabetic polyneuro-pathy,several symptom scoring systems are available, which are often extensive andlack in validation. We validated a new four-item Diabetic Neuropathy Symptom (DNS) scorefor diagnosing distal diabetic polyneuropathy.

  12. A scoring system for ascertainment of incident stroke; the Risk Index Score (RISc).

    Science.gov (United States)

    Kass-Hout, T A; Moyé, L A; Smith, M A; Morgenstern, L B

    2006-01-01

    The main objective of this study was to develop and validate a computer-based statistical algorithm that could be translated into a simple scoring system in order to ascertain incident stroke cases using hospital admission medical records data. The Risk Index Score (RISc) algorithm was developed using data collected prospectively by the Brain Attack Surveillance in Corpus Christi (BASIC) project, 2000. The validity of RISc was evaluated by estimating the concordance of scoring system stroke ascertainment to stroke ascertainment by physician and/or abstractor review of hospital admission records. RISc was developed on 1718 randomly selected patients (training set) and then statistically validated on an independent sample of 858 patients (validation set). A multivariable logistic model was used to develop RISc and subsequently evaluated by goodness-of-fit and receiver operating characteristic (ROC) analyses. The higher the value of RISc, the higher the patient's risk of potential stroke. The study showed RISc was well calibrated and discriminated those who had potential stroke from those that did not on initial screening. In this study we developed and validated a rapid, easy, efficient, and accurate method to ascertain incident stroke cases from routine hospital admission records for epidemiologic investigations. Validation of this scoring system was achieved statistically; however, clinical validation in a community hospital setting is warranted.

  13. Validity of GRE General Test scores and TOEFL scores for graduate admission to a technical university in Western Europe

    Science.gov (United States)

    Zimmermann, Judith; von Davier, Alina A.; Buhmann, Joachim M.; Heinimann, Hans R.

    2018-01-01

    Graduate admission has become a critical process in tertiary education, whereby selecting valid admissions instruments is key. This study assessed the validity of Graduate Record Examination (GRE) General Test scores for admission to Master's programmes at a technical university in Europe. We investigated the indicative value of GRE scores for the Master's programme grade point average (GGPA) with and without the addition of the undergraduate GPA (UGPA) and the TOEFL score, and of GRE scores for study completion and Master's thesis performance. GRE scores explained 20% of the variation in the GGPA, while additional 7% were explained by the TOEFL score and 3% by the UGPA. Contrary to common belief, the GRE quantitative reasoning score showed only little explanatory power. GRE scores were also weakly related to study progress but not to thesis performance. Nevertheless, GRE and TOEFL scores were found to be sensible admissions instruments. Rigorous methodology was used to obtain highly reliable results.

  14. Application of the FOUR Score in Intracerebral Hemorrhage Risk Analysis.

    Science.gov (United States)

    Braksick, Sherri A; Hemphill, J Claude; Mandrekar, Jay; Wijdicks, Eelco F M; Fugate, Jennifer E

    2018-06-01

    The Full Outline of Unresponsiveness (FOUR) Score is a validated scale describing the essentials of a coma examination, including motor response, eye opening and eye movements, brainstem reflexes, and respiratory pattern. We incorporated the FOUR Score into the existing ICH Score and evaluated its accuracy of risk assessment in spontaneous intracerebral hemorrhage (ICH). Consecutive patients admitted to our institution from 2009 to 2012 with spontaneous ICH were reviewed. The ICH Score was calculated using patient age, hemorrhage location, hemorrhage volume, evidence of intraventricular extension, and Glasgow Coma Scale (GCS). The FOUR Score was then incorporated into the ICH Score as a substitute for the GCS (ICH Score FS ). The ability of the 2 scores to predict mortality at 1 month was then compared. In total, 274 patients met the inclusion criteria. The median age was 73 years (interquartile range 60-82) and 138 (50.4%) were male. Overall mortality at 1 month was 28.8% (n = 79). The area under the receiver operating characteristic curve was .91 for the ICH Score and .89 for the ICH Score FS . For ICH Scores of 1, 2, 3, 4, and 5, 1-month mortality was 4.2%, 29.9%, 62.5%, 95.0%, and 100%. In the ICH Score FS model, mortality was 10.7%, 26.5%, 64.5%, 88.9%, and 100% for scores of 1, 2, 3, 4, and 5, respectively. The ICH Score and the ICH Score FS predict 1-month mortality with comparable accuracy. As the FOUR Score provides additional clinical information regarding patient status, it may be a reasonable substitute for the GCS into the ICH Score. Copyright © 2018 National Stroke Association. Published by Elsevier Inc. All rights reserved.

  15. Hospital Value-Based Purchasing (HVBP) – Total Performance Score

    Data.gov (United States)

    U.S. Department of Health & Human Services — A list of hospitals participating in the Hospital VBP Program and their Clinical Process of Care domain scores, Patient Experience of Care dimension scores, and...

  16. Lower Bounds to the Reliabilities of Factor Score Estimators.

    Science.gov (United States)

    Hessen, David J

    2016-10-06

    Under the general common factor model, the reliabilities of factor score estimators might be of more interest than the reliability of the total score (the unweighted sum of item scores). In this paper, lower bounds to the reliabilities of Thurstone's factor score estimators, Bartlett's factor score estimators, and McDonald's factor score estimators are derived and conditions are given under which these lower bounds are equal. The relative performance of the derived lower bounds is studied using classic example data sets. The results show that estimates of the lower bounds to the reliabilities of Thurstone's factor score estimators are greater than or equal to the estimates of the lower bounds to the reliabilities of Bartlett's and McDonald's factor score estimators.

  17. The Truth about Scores Children Achieve on Tests.

    Science.gov (United States)

    Brown, Jonathan R.

    1989-01-01

    The importance of using the standard error of measurement (SEm) in determining reliability in test scores is emphasized. The SEm is compared to the hypothetical true score for standardized tests, and procedures for calculation of the SEm are explained. (JDD)

  18. A locally adapted functional outcome measurement score for total ...

    African Journals Online (AJOL)

    ... in Europe or North America and seem not optimally suited for a general West ... We introduce a cross-cultural adaptation of the Lequesne index as a new score. ... Keywords: THR, Hip, Africa, Functional score, Hip replacement, Arthroscopy ...

  19. 48 CFR 1515.305-70 - Scoring plans.

    Science.gov (United States)

    2010-10-01

    ... METHODS AND CONTRACT TYPES CONTRACTING BY NEGOTIATION Source Selection 1515.305-70 Scoring plans. When... solicitation, e.g., other numeric, adjectival, color rating systems, etc. Scoring Plan Value Descriptive...

  20. Methods and statistics for combining motif match scores.

    Science.gov (United States)

    Bailey, T L; Gribskov, M

    1998-01-01

    Position-specific scoring matrices are useful for representing and searching for protein sequence motifs. A sequence family can often be described by a group of one or more motifs, and an effective search must combine the scores for matching a sequence to each of the motifs in the group. We describe three methods for combining match scores and estimating the statistical significance of the combined scores and evaluate the search quality (classification accuracy) and the accuracy of the estimate of statistical significance of each. The three methods are: 1) sum of scores, 2) sum of reduced variates, 3) product of score p-values. We show that method 3) is superior to the other two methods in both regards, and that combining motif scores indeed gives better search accuracy. The MAST sequence homology search algorithm utilizing the product of p-values scoring method is available for interactive use and downloading at URL http:/(/)www.sdsc.edu/MEME.

  1. Modifying scoring system at South African University rugby level ...

    African Journals Online (AJOL)

    Success in rugby is measured by winning the game and in order to do so, teams need to score more points ... if modifying the scoring system at South African University rugby level changes the game dynamics. ... AJOL African Journals Online.

  2. The Incentive Effect of Scores: Randomized Evidence from Credit Committees

    OpenAIRE

    Daniel Paravisini; Antoinette Schoar

    2013-01-01

    We design a randomized controlled trial to evaluate the adoption of credit scoring with a bank that uses soft information in small businesses lending. We find that credit scores improve the productivity of credit committees, reduce managerial involvement in the loan approval process, and increase the profitability of lending. Credit committee members' effort and output also increase when they anticipate the score becoming available, indicating that scores improve incentives to use existing in...

  3. Proposal of a Mediterranean Diet Serving Score.

    Directory of Open Access Journals (Sweden)

    Celia Monteagudo

    Full Text Available Numerous studies have demonstrated a relationship between Mediterranean Diet (MD adherence and the prevention of cardiovascular diseases, cancer, and diabetes, etc. The study aim was to validate a novel instrument to measure MD adherence based on the consumption of food servings and food groups, and apply it in a female population from southern Spain and determining influential factors.The study included 1,155 women aged 12-83 yrs, classified as adolescents, adults, and over-60-yr-olds. All completed a validated semi-quantitative food frequency questionnaire (FFQ. The Mediterranean Dietary Serving Score (MDSS is based on the latest update of the Mediterranean Diet Pyramid, using the recommended consumption frequency of foods and food groups; the MDSS ranges from 0 to 24. The discriminative power or correct subject classification capacity of the MDSS was analyzed with the Receiver Operating Characteristic (ROC curve, using the MDS as reference method. Predictive factors for higher MDSS adherence were determined with a logistic regression model, adjusting for age. According to ROC curve analysis, MDSS evidenced a significant discriminative capacity between adherents and non-adherents to the MD pattern (optimal cutoff point=13.50; sensitivity=74%; specificity=48%. The mean MDSS was 12.45 (2.69 and was significantly higher with older age (p<0.001. Logistic regression analysis showed highest MD adherence by over 60-year-olds with low BMI and no habit of eating between meals.The MDSS is an updated, easy, valid, and accurate instrument to assess MD adherence based on the consumption of foods and food groups per meal, day, and week. It may be useful in future nutritional education programs to prevent the early onset of chronic non-transmittable diseases in younger populations.

  4. Gambling score in earthquake prediction analysis

    Science.gov (United States)

    Molchan, G.; Romashkova, L.

    2011-03-01

    The number of successes and the space-time alarm rate are commonly used to characterize the strength of an earthquake prediction method and the significance of prediction results. It has been recently suggested to use a new characteristic to evaluate the forecaster's skill, the gambling score (GS), which incorporates the difficulty of guessing each target event by using different weights for different alarms. We expand parametrization of the GS and use the M8 prediction algorithm to illustrate difficulties of the new approach in the analysis of the prediction significance. We show that the level of significance strongly depends (1) on the choice of alarm weights, (2) on the partitioning of the entire alarm volume into component parts and (3) on the accuracy of the spatial rate measure of target events. These tools are at the disposal of the researcher and can affect the significance estimate. Formally, all reasonable GSs discussed here corroborate that the M8 method is non-trivial in the prediction of 8.0 ≤M < 8.5 events because the point estimates of the significance are in the range 0.5-5 per cent. However, the conservative estimate 3.7 per cent based on the number of successes seems preferable owing to two circumstances: (1) it is based on relative values of the spatial rate and hence is more stable and (2) the statistic of successes enables us to construct analytically an upper estimate of the significance taking into account the uncertainty of the spatial rate measure.

  5. Trainee Occupational Therapists Scoring the Barthel ADL.

    Science.gov (United States)

    Martin, Elizabeth; Nugent, Chris; Bond, Raymond; Martin, Suzanne

    2015-09-01

    Within medical applications there are two main types of information design; paper-based and digital information [1]. As technology is constantly changing, information within healthcare management and delivery is continually being transitioned from traditional paper documents to digital and online resources. Activity of Daily Living (ADL) charts are still predominantly paper based and are therefore prone to "human error" [2]. In light of this, an investigation has taken place into the design for reducing the amount of human error, between a paper based ADL, specifically the Barthel Index, and the same ADL created digitally. The digital ADL was developed as an online platform as this offers the best method of data capture for a large group of participants all together [3]. The aim of the study was to evaluate the usability of the Barthel Index ADL in paper format and then reproduce the same ADL digitally. This paper presents the findings of a study involving 26 participants who were familiar with ADL charts, and used three scenarios requiring them to complete both a paper ADL and a digital ADL. An evaluation was undertaken to ascertain if there were any 'human errors' in completing the paper ADL and also to find similarities/differences through using the digital ADL. The results from the study indicated that 22/26 participants agreed that the digital ADL was better, if not the same as a paper based ADL. Further results indicated that participants rate highly the added benefit of the digital ADL being easy to use and also that calculation of assessment scores were performed automatically. Statistically the digital BI offered a 100 % correction rate in the total calculation, in comparison to the paper based BI where it is more common for users to make mathematical calculation errors. Therefore in order to minimise handwriting and calculation errors, the digital BI proved superior than the traditional paper based method.

  6. Dose Uniformity of Scored and Unscored Tablets: Application of the FDA Tablet Scoring Guidance for Industry.

    Science.gov (United States)

    Ciavarella, Anthony B; Khan, Mansoor A; Gupta, Abhay; Faustino, Patrick J

    This U.S. Food and Drug Administration (FDA) laboratory study examines the impact of tablet splitting, the effect of tablet splitters, and the presence of a tablet score on the dose uniformity of two model drugs. Whole tablets were purchased from five manufacturers for amlodipine and six for gabapentin. Two splitters were used for each drug product, and the gabapentin tablets were also split by hand. Whole and split amlodipine tablets were tested for content uniformity following the general chapter of the United States Pharmacopeia (USP) Uniformity of Dosage Units , which is a requirement of the new FDA Guidance for Industry on tablet scoring. The USP weight variation method was used for gabapentin split tablets based on the recommendation of the guidance. All whole tablets met the USP acceptance criteria for the Uniformity of Dosage Units. Variation in whole tablet content ranged from 0.5 to 2.1 standard deviation (SD) of the percent label claim. Splitting the unscored amlodipine tablets resulted in a significant increase in dose variability of 6.5-25.4 SD when compared to whole tablets. Split tablets from all amlodipine drug products did not meet the USP acceptance criteria for content uniformity. Variation in the weight for gabapentin split tablets was greater than the whole tablets, ranging from 1.3 to 9.3 SD. All fully scored gabapentin products met the USP acceptance criteria for weight variation. Size, shape, and the presence or absence of a tablet score can affect the content uniformity and weight variation of amlodipine and gabapentin tablets. Tablet splitting produced higher variability. Differences in dose variability and fragmentation were observed between tablet splitters and hand splitting. These results are consistent with the FDA's concerns that tablet splitting can have an effect on the amount of drug present in a split tablet and available for absorption. Tablet splitting has become a very common practice in the United States and throughout the

  7. Comparing the Scoring of Human Decomposition from Digital Images to Scoring Using On-site Observations.

    Science.gov (United States)

    Dabbs, Gretchen R; Bytheway, Joan A; Connor, Melissa

    2017-09-01

    When in forensic casework or empirical research in-person assessment of human decomposition is not possible, the sensible substitution is color photographic images. To date, no research has confirmed the utility of color photographic images as a proxy for in situ observation of the level of decomposition. Sixteen observers scored photographs of 13 human cadavers in varying decomposition stages (PMI 2-186 days) using the Total Body Score system (total n = 929 observations). The on-site TBS was compared with recorded observations from digital color images using a paired samples t-test. The average difference between on-site and photographic observations was -0.20 (t = -1.679, df = 928, p = 0.094). Individually, only two observers, both students with human decomposition based on digital images can be substituted for assessments based on observation of the corpse in situ, when necessary. © 2017 American Academy of Forensic Sciences.

  8. Direct power comparisons between simple LOD scores and NPL scores for linkage analysis in complex diseases.

    Science.gov (United States)

    Abreu, P C; Greenberg, D A; Hodge, S E

    1999-09-01

    Several methods have been proposed for linkage analysis of complex traits with unknown mode of inheritance. These methods include the LOD score maximized over disease models (MMLS) and the "nonparametric" linkage (NPL) statistic. In previous work, we evaluated the increase of type I error when maximizing over two or more genetic models, and we compared the power of MMLS to detect linkage, in a number of complex modes of inheritance, with analysis assuming the true model. In the present study, we compare MMLS and NPL directly. We simulated 100 data sets with 20 families each, using 26 generating models: (1) 4 intermediate models (penetrance of heterozygote between that of the two homozygotes); (2) 6 two-locus additive models; and (3) 16 two-locus heterogeneity models (admixture alpha = 1.0,.7,.5, and.3; alpha = 1.0 replicates simple Mendelian models). For LOD scores, we assumed dominant and recessive inheritance with 50% penetrance. We took the higher of the two maximum LOD scores and subtracted 0.3 to correct for multiple tests (MMLS-C). We compared expected maximum LOD scores and power, using MMLS-C and NPL as well as the true model. Since NPL uses only the affected family members, we also performed an affecteds-only analysis using MMLS-C. The MMLS-C was both uniformly more powerful than NPL for most cases we examined, except when linkage information was low, and close to the results for the true model under locus heterogeneity. We still found better power for the MMLS-C compared with NPL in affecteds-only analysis. The results show that use of two simple modes of inheritance at a fixed penetrance can have more power than NPL when the trait mode of inheritance is complex and when there is heterogeneity in the data set.

  9. Nutech functional score: A novel scoring system to assess spinal cord injury patients.

    Science.gov (United States)

    Shroff, Geeta; Barthakur, Jitendra Kumar

    2017-06-26

    To develop a new scoring system, nutech functional scores (NFS) for assessing the patients with spinal cord injury (SCI). The conventional scale, American Spinal Injury Association's (ASIA) impairment scale is a measure which precisely describes the severity of the SCI. However, it has various limitations which lead to incomplete assessment of SCI patients. We have developed a 63 point scoring system, i . e ., NFS for patients suffering with SCI. A list of symptoms either common or rare that were found to be associated with SCI was recorded for each patient. On the basis of these lists, we have developed NFS. These lists served as a base to prepare NFS, a 63 point positional (each symptom is sub-graded and get points based on position) and directional (moves in direction BAD → GOOD) scoring system. For non-progressive diseases, 1, 2, 3, 4, 5 denote worst, bad, moderate, good and best (normal), respectively. NFS for SCI has been divided into different groups based on the affected part of the body being assessed, i . e ., motor assessment (shoulders, elbow, wrist, fingers-grasp, fingers-release, hip, knee, ankle and toe), sensory assessment, autonomic assessment, bed sore assessment and general assessment. As probability based studies required a range of (-1, 1) or at least the range of (0, 1) to be useful for real world analysis, the grades were converted to respective numeric values. NFS can be considered as a unique tool to assess the improvement in patients with SCI as it overcomes the limitations of ASIA impairment scale.

  10. Severity scoring in the critically ill: part 2: maximizing value from outcome prediction scoring systems.

    Science.gov (United States)

    Breslow, Michael J; Badawi, Omar

    2012-02-01

    Part 2 of this review of ICU scoring systems examines how scoring system data should be used to assess ICU performance. There often are two different consumers of these data: lCU clinicians and quality leaders who seek to identify opportunities to improve quality of care and operational efficiency, and regulators, payors, and consumers who want to compare performance across facilities. The former need to know how to garner maximal insight into their care practices; this includes understanding how length of stay (LOS) relates to quality, analyzing the behavior of different subpopulations, and following trends over time. Segregating patients into low-, medium-, and high-risk populations is especially helpful, because care issues and outcomes may differ across this severity continuum. Also, LOS behaves paradoxically in high-risk patients (survivors often have longer LOS than nonsurvivors); failure to examine this subgroup separately can penalize ICUs with superior outcomes. Consumers of benchmarking data often focus on a single score, the standardized mortality ratio (SMR). However, simple SMRs are disproportionately affected by outcomes in high-risk patients, and differences in population composition, even when performance is otherwise identical, can result in different SMRs. Future benchmarking must incorporate strategies to adjust for differences in population composition and report performance separately for low-, medium- and high-acuity patients. Moreover, because many ICUs lack the resources to care for high-acuity patients (predicted mortality >50%), decisions about where patients should receive care must consider both ICU performance scores and their capacity to care for different types of patients.

  11. Improving personality facet scores with multidimensional computer adaptive testing

    DEFF Research Database (Denmark)

    Makransky, Guido; Mortensen, Erik Lykke; Glas, Cees A W

    2013-01-01

    personality tests contain many highly correlated facets. This article investigates the possibility of increasing the precision of the NEO PI-R facet scores by scoring items with multidimensional item response theory and by efficiently administering and scoring items with multidimensional computer adaptive...

  12. Classifying snakebite in South Africa: Validating a scoring system ...

    African Journals Online (AJOL)

    Factors predictive of ATI and the optimal cut-off score for predicting an ATI were identified. These factors were then used to develop a standard scoring system. The score was then tested prospectively for accuracy in a new validation cohort consisting of 100 patients admitted for snakebite to our unit from 1 December 2014 to ...

  13. The comparison of modified early warning score and Glasgow coma ...

    African Journals Online (AJOL)

    Introduction: The purpose of this study is to assess and compare the discriminatory ability of the Glasgow coma scale (GCS)‑age‑systolic blood pressure (GAP) score and modified early warning scoring system (mEWS) score for 4‑week mortality, for the patients being in the triage category 1 and 2 who refer to Emergency ...

  14. a locally adapted functional outcome measurement score for total

    African Journals Online (AJOL)

    Results and success of total hip arthroplasty are often measured using a functional outcome scoring system. Most current scores were developed in Europe and. North America (1-3). During the evaluation of a Total. Hip Replacement (THR) project in Ouagadougou,. Burkina Faso (4) it was felt that these scores were not.

  15. Family Functioning and Child Psychopathology: Individual Versus Composite Family Scores.

    Science.gov (United States)

    Mathijssen, Jolanda J. J. P.; Koot, Hans M.; Verhulst, Frank C.; De Bruyn, Eric E. J.; Oud, Johan H. L.

    1997-01-01

    Examines the relationship of individual family members' perceptions and family mean and discrepancy scores of cohesion and adaptability with child psychopathology in a sample of 138 families. Results indicate that family mean scores, contrary to family discrepancy scores, explain more of the variance in parent-reported child psychopathology than…

  16. LCA single score analysis of man-made cellulose fibres

    NARCIS (Netherlands)

    Shen, L.; Patel, M.K.

    2010-01-01

    In this study, the LCA report “Life Cycle assessment of man-made cellulose fibres” [3] is extended to the single score analysis in order to provide an additional basis for decision making. The single score analysis covers 9 to 11 environmental impact categories. Three single score methods (Single

  17. A Framework for Evaluation and Use of Automated Scoring

    Science.gov (United States)

    Williamson, David M.; Xi, Xiaoming; Breyer, F. Jay

    2012-01-01

    A framework for evaluation and use of automated scoring of constructed-response tasks is provided that entails both evaluation of automated scoring as well as guidelines for implementation and maintenance in the context of constantly evolving technologies. Consideration of validity issues and challenges associated with automated scoring are…

  18. Personality and Examination Score Correlates of Abnormal Psychology Course Ratings.

    Science.gov (United States)

    Pauker, Jerome D.

    The relationship between the ratings students assigned to an evening undergraduate abnormal psychology class and their scores on objective personality tests and course examinations was investigated. Students (N=70) completed the MMPI and made global ratings of the course; these scores were correlated separately by sex with the T scores of 13 MMPI…

  19. Confidence Scoring of Speaking Performance: How Does Fuzziness become Exact?

    Science.gov (United States)

    Jin, Tan; Mak, Barley; Zhou, Pei

    2012-01-01

    The fuzziness of assessing second language speaking performance raises two difficulties in scoring speaking performance: "indistinction between adjacent levels" and "overlap between scales". To address these two problems, this article proposes a new approach, "confidence scoring", to deal with such fuzziness, leading to "confidence" scores between…

  20. Clinical outcome scoring of intra-articular calcaneal fractures

    NARCIS (Netherlands)

    Schepers, Tim; Heetveld, Martin J.; Mulder, Paul G. H.; Patka, Peter

    2008-01-01

    Outcome reporting of intra-articular calcaneal fractures is inconsistent. This study aimed to identify the most cited outcome scores in the literature and to analyze their reliability and validity. A systematic literature search identified 34 different outcome scores. The most cited outcome score