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Sample records for electrophoresis comet assays

  1. Standardizing electrophoresis conditions: how to eliminate a major source of error in the comet assay.

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    Gunnar Brunborg

    2015-06-01

    Full Text Available In the alkaline comet assay, cells are embedded in agarose, lysed, and then subjected to further processing including electrophoresis at high pH (>13. We observed very large variations of mean comet tail lengths of cell samples from the same population when spread on a glass or plastic substrate and subjected to electrophoresis. These variations might be cancelled out if comets are scored randomly over a large surface, or if all the comets are scored. The mean tail length may then be representative of the population, although its standard error is large. However, the scoring process often involves selection of 50 – 100 comets in areas selected in an unsystematic way from a large gel on a glass slide. When using our 96-sample minigel format (1, neighbouring sample variations are easily detected. We have used this system to study the cause of the comet assay variations during electrophoresis and we have defined experimental conditions which reduce the variations to a minimum. We studied the importance of various physical parameters during electrophoresis: (i voltage; (ii duration of electrophoresis; (iii electric current; (iv temperature; and (v agarose concentration. We observed that the voltage (V/cm varied substantially during electrophoresis, even within a few millimetres of distance between gel samples. Not unexpectedly, both the potential ( V/cm and the time were linearly related to the mean comet tail, whereas the current was not. By measuring the local voltage with microelectrodes a few millimetres apart, we observed substantial local variations in V/cm, and they increased with time. This explains the large variations in neighbouring sample comet tails of 25% or more. By introducing simple technology (circulation of the solution during electrophoresis, and temperature control, these variations in mean comet tail were largely abolished, as were the V/cm variations. Circulation was shown to be particularly important and optimal conditions

  2. Assessment of individual radiosensitivity in human lymphocytes using micronucleus and microgel electrophoresis Comet assays

    Energy Technology Data Exchange (ETDEWEB)

    Giorgio, M. di; Sardi, M.; Busto, M.; Vallerga, M.; Taja, M.; Mairal, I.

    2004-07-01

    Background and purpose: Individual radiosensitivity is an inherent characteristic, associated with an increased reaction to ionizing radiation on the human body. Individuals show marked differences in radiation sensitivity, which has consequences in the fields of both radiation protection and radiation therapy. It is suggested that DNA repair mechanisms are involved. Consequently, the characterization of DNA repair in lymphocytes through cytokinesis blocked micronucleus (MN) and alkaline single-cell microgel electrophoresis (comet) assays could be suitable approaches to evaluate individual radiosensitivity in vitro. The amins of this study were: 1) to assess the in vitro radisensitivity of peripheral blood lymphocytes from two with the observed clinical response and 2) to test the predictive potential of both techniques. Materials and methods: 38 cancer patients receiving radiation therapy were enrolled in this study. The tumor sites were: head and neck (n=25) and cervic (n=13). 19 pateints were evaluated prior, mid-way and on completion of treatment (prospective group) and 19 patients were evaluated about 2-480 month after radiotherapy (retrospective group). Cytogenetic data from the prospective group were analyzed 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 cytogentic recovery factor k. In the retrospective group, blood samples were irradiated in vitro with 0 (control) or 2 Gy and evaluated using MN test. Cytogenetic data were analyzed comparing expected MN frequencies (calibration curve from health donors) with values observed after in vitro irradiation. One over-reactor ad patients that did not develop late effects were also evaluated through comet assay. DNA damage and repair capacity were quantified by the Olive tail moment. Lymphocytes of health individuals were used as reference sample. In the prospective evaluation, factor K correlated

  3. Assessment of individual radiosensitivity in human lymphocytes using micronucleus and microgel electrophoresis Comet assays

    International Nuclear Information System (INIS)

    Giorgio, M. di; Sardi, M.; Busto, M.; Vallerga, M.; Taja, M.; Mairal, I.

    2004-01-01

    Background and purpose: Individual radiosensitivity is an inherent characteristic, associated with an increased reaction to ionizing radiation on the human body. Individuals show marked differences in radiation sensitivity, which has consequences in the fields of both radiation protection and radiation therapy. It is suggested that DNA repair mechanisms are involved. Consequently, the characterization of DNA repair in lymphocytes through cytokinesis blocked micronucleus (MN) and alkaline single-cell microgel electrophoresis (comet) assays could be suitable approaches to evaluate individual radiosensitivity in vitro. The amins of this study were: 1) to assess the in vitro radisensitivity of peripheral blood lymphocytes from two with the observed clinical response and 2) to test the predictive potential of both techniques. Materials and methods: 38 cancer patients receiving radiation therapy were enrolled in this study. The tumor sites were: head and neck (n=25) and cervic (n=13). 19 pateints were evaluated prior, mid-way and on completion of treatment (prospective group) and 19 patients were evaluated about 2-480 month after radiotherapy (retrospective group). Cytogenetic data from the prospective group were analyzed 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 cytogentic recovery factor k. In the retrospective group, blood samples were irradiated in vitro with 0 (control) or 2 Gy and evaluated using MN test. Cytogenetic data were analyzed comparing expected MN frequencies (calibration curve from health donors) with values observed after in vitro irradiation. One over-reactor ad patients that did not develop late effects were also evaluated through comet assay. DNA damage and repair capacity were quantified by the Olive tail moment. Lymphocytes of health individuals were used as reference sample. In the prospective evaluation, factor K correlated

  4. ASTRO Research Fellow Presentation - A comparison of the comet assay and pulsed-field gel electrophoresis as a predictive assay for radiosensitivity in human fibroblasts

    International Nuclear Information System (INIS)

    Sarkaria, Jann N.; Eady, John J.; Peacock, John H.; Steel, G. Gordon

    1996-01-01

    Purpose/Objective: To determine whether neutral lysis single-cell gel electrophoresis (comet assay) or pulsed-field gel electrophoresis (PFGE) can be used as a predictive assay for tissue response to radiotherapy as an alternative to clonogenic survival measurements. Materials and Methods: The comet assay has been widely used to measure DNA double strand breaks (dsb) in individual cells, and it has been suggested that it could be used as an alternative to clonogenic assays to measure radiosensitivity. Previous studies in this lab have demonstrated the ability of pulsed-field gel electrophoresis, which also measures DNA dsb, to accurately predict the radiosensitivity of a panel of fibroblasts based on determination of residual DNA dsb. As part of an ongoing study examining the relationship between fibroblast radiosensitivity and normal-tissue radiation reactions, we have compared the sensitivity and accuracy of the comet assay and PFGE on a different panel of non-transformed fibroblasts derived from breast cancer patients who developed severe radiation late effects and from case-matched controls. For the measurement of initial damage, cells were suspended in PBS and irradiated on ice for the comet assay and irradiated in agarose plugs on ice for pFGE. Residual damage was measured following irradiation of confluent cultures at 37 degree sign C and subsequent incubation for four hours prior to preparation of agarose slides and plugs. All irradiations were performed with a 59 TBq 60 Co source at a dose rate of 1.7 Gy/min. Electrophoresis was performed following neutral pH cell lysis. Comet images were captured and analyzed using Optimas software with DNA damage quantitated by the comet moment. PFGE gels were analyzed using a phosphor-image analysis system and damage was quantitated based on the percent of activity released from the well. Results: The comet assay was able to detect initial DNA damage at a threshold of 5 Gy and exhibited a linear dose

  5. Viable human buccal mucosa cells do not yield typical nucleoids: impacts on the single-cell gel electrophoresis/Comet assay.

    Science.gov (United States)

    Pinhal, Danillo; Gontijo, Alisson Marques de Miranda Cabral; Reyes, Victor Alexis Valenzuela; Salvadori, Daisy Maria Favero

    2006-03-01

    Buccal mucosa (BM) cells have been used in human biomonitoring studies for detecting DNA adducts and chromosomal damage in an epithelial cell population. In the present study, we have investigated if human BM cells are suitable for use in the single-cell gel electrophoresis (SCGE)/Comet assay as an approach for estimating the exposure of epithelial cells to DNA-damaging agents. Our results indicate that only a few cells from BM cell samples yield comets that can be analyzed by current methods, and that the yield of cells with comets is independent of the percentage of viable BM cells in the sample. Data generated after enzymatic enrichment of viable cells and immunomagnetic separation of epithelial cells suggest that most of the BM cells that do form comets are probably leukocytes. Moreover, by reevaluating specific cells after running the Comet assay, we found that viable epithelial BM cells give rise to atypical comets that are not included in the analysis. Comparing DNA migration patterns between small groups of smokers and nonsmokers indicated that long-term smoking had no effect on the subpopulation of cells that yield typical comets. Our results indicate that the SCGE assay, as it is commonly performed, may not be useful for genotoxicity monitoring in human epithelial BM cells. (c) 2005 Wiley-Liss, Inc.

  6. Evaluation of radio-induced DNA damage and their repair in human lymphocytes by comet assay or single cell gel electrophoresis

    International Nuclear Information System (INIS)

    Nascimento, Patricia A. do; Suzuki, Miriam F.; Okazaki, Kayo

    1997-01-01

    The comet assay, also called single cell gel electrophoresis technique, permits to evaluate quantitatively DNA breakage induced by chemical and physical agents at the level of the single cell. The present paper refers to the construction of dose-response curves to DNA damage and repair studies in human peripheral lymphocytes, utilizing the comet assay for the radiosensitivity analysis. So, the blood samples were obtained from healthy donors (40-50 year old), irradiated in a 60 Co source (GAMMACEL 220) with doses of 0.17, 0.25, 0.57, 1.10, 2.12 and 4.22 Gy (0.59 Gy/min.) and processed 1 and 24 hours after the exposition. Results obtained showed a increase in the total lenght of comet (DNA migration) as a function of radiation dose in samples processed 1 and 24 hours after the treatment. The DNA lesion in irradiated lymphocytes with 4.22 Gy (means value of 101.4 μm) were 3.4 times higher than in the untreated lymphocytes (mean value of 30 μm) instead of 24 hours after the irradiation were 1.5 times higher (mean value of 46.3 μm). This reduction on DNA repair occurred in these cells. It was also possible visualized the presence of subpopulations of the cells with different sensitivity and repair capacity to ionizing radiation in these donors. (author). 8 refs., 3 figs

  7. Assessment and reduction of comet assay variation in relation to DNA damage: studies from the European Comet Assay Validation Group

    DEFF Research Database (Denmark)

    Møller, Peter; Möller, Lennart; Godschalk, Roger W L

    2010-01-01

    The alkaline single cell gel electrophoresis (comet) assay has become a widely used method for the detection of DNA damage and repair in cells and tissues. Still, it has been difficult to compare results from different investigators because of differences in assay conditions and because the data...... are reported in different units. The European Comet Assay Validation Group (ECVAG) was established for the purpose of validation of the comet assay with respect to measures of DNA damage formation and its repair. The results from this inter-laboratory validation trail showed a large variation in measured level...... reliability for the measurement of DNA damage by the comet assay but there is still a need for further validation to reduce both assay and inter-laboratory variation....

  8. Validation of comet assay using different genotoxicity tests in mammals

    International Nuclear Information System (INIS)

    Balid, Arundhati; Godbole, N.N.; Bhilwade, H.N.; Rajagopalan, R.; Chaubey, R.C.

    2004-01-01

    Present studies were undertaken to compare the sensitivity of three test systems: chromosome aberration test, micronucleus test and single cell gel electrophoresis/comet assay. These tests were carried out in a single animal in order to optimize, evaluate and compare their relative sensitivities

  9. Controlling variation in the comet assay

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    Andrew Richard Collins

    2014-10-01

    Full Text Available Variability of the comet assay is a serious issue, whether it occurs from experiment to experiment in the same laboratory, or between different laboratories analysing identical samples. Do we have to live with high variability, just because the comet assay is a biological assay rather than analytical chemistry? Numerous attempts have been made to limit variability by standardising the assay protocol, and the critical steps in the assay have been identified; agarose concentration, duration of alkaline incubation, and electrophoresis conditions (time, temperature and voltage gradient are particularly important. Even when these are controlled, variation seems to be inevitable. It is helpful to include in experiments reference standards, i.e. cells with a known amount of specific damage to the DNA. They can be aliquots frozen from a single large batch of cells, either untreated (negative controls or treated with, for example, H2O2 or X-rays to induce strand breaks (positive control for the basic assay, or photosensitiser plus light to oxidise guanine (positive control for Fpg- or OGG1-sensitive sites. Reference standards are especially valuable when performing a series of experiments over a long period - for example, analysing samples of white blood cells from a large human biomonitoring trial - to check that the assay is performing consistently, and to identify anomalous results necessitating a repeat experiment. The reference values of tail intensity can also be used to iron out small variations occurring from day to day. We present examples of the use of reference standards in human trials, both within one laboratory and between different laboratories, and describe procedures that can be used to control variation.

  10. The application of single cell gel electrophoresis or comet assay to human monitoring studies Aplicacion de la electroforesis unicelular o ensayo cometa en estudios de monitoreo humano

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    Mahara Valverde

    1999-11-01

    Full Text Available Objective. In the search of new human genotoxic biomarkers, the single cell gel electrophoresis assay has been proposed as a sensible alternative. Material and methods. This technique detects principally single strand breaks as well as alkali-labile and repair-retarded sites. Results. Herein we present our experience using the single cell gel electrophoresis assay in human population studies, both occupationally and environmentally exposed. Conclusions. We discuss the assay feasibility as a genotoxic biomarker.Objetivo. En la búsqueda de nuevos marcadores genotóxicos aplicables a estudios de poblaciones humanas expuestos a xenobióticos, la utilización del ensayo de electroforesis en una sola célula se ha propuesto como un método sensible y una buena alternativa. Material y métodos. Esta técnica detecta rompimientos en el ADN de cadena sencilla, así como sitios álcali lábiles y sitios retardados de reparación. Resultados. En este trabajo, presentamos nuestra experiencia utilizando este ensayo en poblaciones humanas expuestas ocupacionalmente o ambientalmente a diferentes xenobióticos. Conclusiones. Se discute la posible utilidad de este ensayo como un biomarcador de efecto genotóxico.

  11. Comet assay on mice testicular cells

    DEFF Research Database (Denmark)

    Sharma, Anoop Kumar

    2015-01-01

    of the comet assay in testicular cells. The chemicals included environmentally relevant substances such as Bisphenol A, PFOS and Tetrabrombisphenol A. Statistical power calculations will be presented to aid in the design of future Comet assay studies on testicular cells. Power curves were provided...

  12. Localized irradiations, evaluation through 'Comet Assay'

    International Nuclear Information System (INIS)

    Di Giorgio, Marina; Taja, Maria R.; Nasazzi, Nora B.; Bustos, N.; Cavalieri, H.; Bolgiani, A.

    2000-01-01

    During the last 50 years various radiation accidents involving localized irradiations occurred, resulting mainly from improper handling of sealed sources of Cobalt 60, Cesium 137 or Iridium 192 at work placed for industrial gammagraphy and other radiation sources. Severe skin reaction may developed at the contact sites. Such inhomogeneous irradiations lead to a differential exposure of lymphocytes in lymphatic tissues or other organs that may recirculate into the peripheral blood producing a mixed irradiated and unirradiated population of lymphocytes. Applying the mathematical models 'Contaminated Poisson' of Dolphin and Qdr method of Sasaki, a mean dose in the irradiated body area and its size can be estimated from unstable chromosome aberration scoring. There are also different biophysical techniques that can give response in localized irradiations. Biological dosimetry is a necessary complement to physical and clinical dosimetries. Thus, there is increasing interest in the assessment of biological markers that permit the detection of radiation induced damage in the localized irradiations. The 'Comet Assay' (single cell gel electrophoresis) is a sensitive, rapid and relatively inexpensive method for measuring DNA damage in individual cells. Single cells are embedded in agarose on microscope slides, lysed to remove the majority of the proteins, electrophoresed, then stained with ethidium bromide in order to visualize the DNA. When visualized using a fluorescent microscope, DNA of undamaged cells appears as a spherical mass occupying the cavity formed by the lysed cell. Following radiation damage, the smaller the fragment size and the grater the number of fragments of DNA, the grater the percentage of DNA that it is able to migrate in an electric field, forming a comet image. The assay can be performed under alkaline conditions to examine DNA single strand breaks (SSBs), or in non denaturing (neutral) conditions to measure double strand breaks (DSBs) in individual

  13. Irradiation detection of food by DNA Comet Assay

    International Nuclear Information System (INIS)

    Khan, A.A.; Delincee, H.

    1999-01-01

    Microgel electrophoresis of single cells or nuclei (DNA Comet Assay) has been investigated to detect irradiation treatment of more than 50 food commodities e.g. meats, seafood, cereals, pulses, nuts, fruits and vegetables, and spices. The foodstuffs have been exposed to radiation doses covering the range of potential commercial irradiation for inactivation of pathogenic and spoilage micro-organisms, for insect disinfestation and for shelf-life extension. The Comet Assay is based on detection of DNA fragments presumptive to irradiation. For most of the food items investigated, the assay can be applied successfully for irradiation detection by working out different conditions of the assay. However, with some of the foods difficulties arose due to - lack of discrimination between the irradiated and unirradiated food samples due to the presence of the same kinds of comets in both cases and the total absence of the typical intact cells in unirradiated samples. - Sufficient DNA material was not available from some of the foods. - Insufficient lysis of the cell walls in case of some plant foods. In conclusion, the DNA Comet Assay can help to detect the irradiation treatment of several varieties of foods using low-cost equipment in a short time of analysis. (orig.)

  14. Comet assay on mice testicular cells

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    Anoop Kumar Sharma

    2015-05-01

    Full Text Available Heritable mutations may result in a variety of adverse outcomes including genetic disease in the offspring. In recent years the focus on germ cell mutagenicity has increased and the “Globally Harmonized System of Classification and Labelling of Chemicals (GHS” has published classification criteria for germ cell mutagens (Speit et al., 2009. The in vivo Comet assay is considered a useful tool for investigating germ cell genotoxicity. In the present study DNA strand breaks in testicular cells of mice were investigated. Different classes of chemicals were tested in order to evaluate the sensitivity of the comet assay in testicular cells. The chemicals included environmentally relevant substances such as Bisphenol A, PFOS and Tetrabrombisphenol A. Statistical power calculations will be presented to aid in the design of future Comet assay studies on testicular cells. Power curves were provided with different fold changes in % tail DNA, different number of cells scored and different number of gels (Hansen et al., 2014. An example is shown in Figure 1. A high throughput version of the Comet assay was used. Samples were scored with a fully automatic comet assay scoring system that provided faster scoring of randomly selected cells.

  15. Evaluation of irradiation in foods using DNA comet assay

    International Nuclear Information System (INIS)

    Khawar, Affaf; Bhatti, Ijaz Ahmad; Khan, Q.M.; Ali, T.; Khan, A.I.; Asi, M.R.

    2011-01-01

    Comet assay is a rapid, inexpensive and sensitive biological technique to detect DNA damage in food stuffs by irradiation. In this study the Comet assay is applied on foods of plant and animal origins. Samples were irradiated by using 60 Co gamma-radiation source. The applied doses were 2, 6 and 10 kGy for food of plant origin and 0.5, 1 and 2 kGy for meat items. The un-irradiated and irradiated samples were clearly differentiated on the basis of DNA fragmentation. During the electrophoresis study, it was found that in un-irradiated cells DNA remained intact and appeared as Comets without tail whereas in irradiated cells Comets with tails were visible due to stretching of fragmented DNA. Moreover, it was also revealed that the DNA tail length was dose dependent. Dry food stuffs (seeds) showed good results as compared to moist foods (meat, fruits and vegetables) due to the absence of background damage. (author)

  16. Comet assay. Pt.1. Theory and practice

    International Nuclear Information System (INIS)

    Kruszewski, M.; Wojewodzka, M.; Iwanenko, T.

    1996-01-01

    Comet assay is a new method for measuring DNA breakage in a single cell. The main applications of the method are estimation of DNA single and double strand breaks, oxidative damage, pyrimidine dimers and (6-4)photoproducts, DNA-DNA and DNA-protein crosslinks. The method is used for studying DNA damage and its repair. (author).19 refs, 9 figs

  17. Ecogenotoxicity testing of aquatic environment by comet assay in plants

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    Anita Mukherjee

    2015-05-01

    Full Text Available One of the goals of environmental monitoring is the detection of potentially hazardous compounds in water. We have set up a standard method to apply the Comet assay in aquatic plants that could be of great interest to evaluate cytotoxicity, genotoxicity and oxidative stress on the same species regarded as most sensitive to environmental pollutants. The aim of the present study was to set up of standardized procedure to evaluate genotoxicity in aquatic plants- Ceratophyllum demersum one that is submerged free floating and the other is Lemna minor - a fresh water floating plant by Comet assay. Electrophoresis and unwinding times were adapted to obtain minimum DNA migration evaluated as tail intensity % or tail moment in the control group and, at the same time maximum sensitivity for DNA damage with known genotoxicants. We further investigated the cytotoxicity and oxidative stress induced in the same species. Based on the repeatability of results obtained we suggest that Ceratophyllum, Lemna can serve as model species and Comet assay could be adopted to monitor the eco-genotoxicity of water pollutants.

  18. Capillary Electrophoresis Analysis of Conventional Splicing Assays

    DEFF Research Database (Denmark)

    de Garibay, Gorka Ruiz; Acedo, Alberto; García-Casado, Zaida

    2014-01-01

    of these assays is often challenging. Here, we explore this issue by conducting splicing assays in 31 BRCA2 genetic variants. All variants were assessed by RT-PCR followed by capillary electrophoresis and direct sequencing. If assays did not produce clear-cut outputs (Class-2 or Class-5 according to analytical...

  19. Comet assay on tetraploid yeast cells

    DEFF Research Database (Denmark)

    Rank, Jette; Syberg, Kristian; Jensen, Klara

    2009-01-01

    Tetraploid yeast cells (Saccharomyces cerevisiae) were used in the comet assay with the intention of developing a new, fast and easy assay for detecting environmental genotoxic agents without using higher organisms. Two DNA-damaging chemicals, H2O2 and acrylamide, together with wastewater from....... Analytical problems that arose due to the small amount of DNA in the yeast nuclei in haploid and diploid cells, which contain 13 Mbp and 26 Mbp DNA per cell, respectively, were solved by using tetraploid yeast cells (52 Mbp) instead. DNA damage was shown after exposure to H2O2 and acrylamide. The lowest dose...... causing significant DNA damage was 20 μM for H2O2 and 200 mg/l for acrylamide. Tertiary-treated wastewater from the outlets of three municipal wastewater-treatment plants was tested, but did not cause DNA damage. Even though it is possible to produce comets with tetraploid yeast cells, the amount of DNA...

  20. Identification of irradiated pepper with comet assay

    International Nuclear Information System (INIS)

    Prieto Miranda, Enrique Fco.; Moreno Alvarez, Damaris L.; Carro Palacio, Sandra; Iglesia Enriquez, Isora

    2007-01-01

    The treatment of foods with ionizing radiations is a technological process utilized in order to increase the hygienic quality and the storage time of the foods. Several methods of detection of irradiated foods have been recommended. The comet assay of DNA is one fast and economical technique for the qualitative identification of irradiated foods. The objective of the present paper was to identify with the comet assay technique the modifications of the DNA molecule of irradiated pepper storage at environment and refrigeration temperatures and different post-irradiation times for different absorbed dose values, (0.1, 0.3 and 0.5 kGy). It was demonstrated that for the high absorbed dose values was observed a greater break into fragments of the DNA molecule, which shows the application of this technique for the identification of irradiated foods. (author)

  1. The Comet Assay: Tails of the (Unexpected. Use of the comet assay in pharmaceutical development.

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    Bas-jan Van Der Leede

    2015-08-01

    Full Text Available In genotoxicity testing of pharmaceuticals the rodent alkaline comet assay is being increasingly used as a second in vivo assay in addition to the in vivo micronucleus assay to mitigate in vitro positive results as recommended by regulatory guidance. In this presentation we want to give insight into the circumstances in vivo comet assay is deployed in a Genetic Toxicology Department of a pharmaceutical company. As the in vivo comet assay is a salvage assay, it means that some events have occurred in an in vitro assay and that the compound (or metabolite responsible for this signal is potentially deselected for further development. More than often the decision to perform an in vivo comet assay is at a very early stage in development and the first time that the compound will be tested in vivo at high/toxic dose levels. As almost no toxicokinetic data and tissue distribution data are available a careful design with maximizes the chances for successful mitigation is necessary. Decisions on acute or repeated dosing need to be made and arrangements for combining the in vivo comet assay with the in vivo micronucleus assay are to be considered. Often synthesis methods need to be scaled up fast to provide the required amount of compound and information on suitable formulations needs to be in place. As exposure data is crucial for interpretation of results, analytical methods need to be brought in place rapidly. An experienced multi skilled and communicative team needs to be available to deploy successfully this kind of assays at an early stage of development. We will present a few scenarios on study conduct and demonstrate how this assay can make a difference for the further development of a new drug.

  2. The JaCVAM / OECD activities on the comet assay

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    Hajime Kojima

    2015-04-01

    Full Text Available The in vivo alkaline single cell gel electrophoresis assay, also called alkaline comet assay is a method measuring DNA strand breaks in eukaryotic cells. This assay was adopted in the Organisation for Economic Co-operation and Development (OECD Test guideline (TG 489 on September 26, 2014. This TG is part of a series of TGs on genetic toxicology. A formal validation trial of the this assay was performed in 2006-2012, coordinated by the Japanese Center for the Validation of Alternative Methods (JaCVAM, in conjunction with the European Union Reference Laboratory for alternatives to animal testing (EURL ECVAM, the Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM and the NTP Interagency Center for the Evaluation of Alternative Toxicological Methods (NICEATM . The assay was reviewed by the OECD genotoxicity experts based on the JaCVAM trial (2014 and in Rothfuss et al. (2010. This TG includes the recommended use and limitations of the comet assay, and is based on the final protocol used in the validation trial, and on additional relevant published and unpublished (laboratories proprietary data. The outline of this TG describes below: each treated group is composed of a minimum of 5 animals of one sex (or of each sex as appropriate. A positive and a vehicle control group are also used. Administration of the treatment consists of daily doses over duration of 2 days or more, ensuring the test chemical reaches the target tissue which can be the liver, the kidney or other tissues if justified. Tissues of interest are dissected and single cells/nuclei suspensions are prepared and embedded in agarose on slides. Cells/nuclei are treated with lysis buffer to remove cellular and/or nuclear membranes. The nuclear DNA in the agar is then subjected to electrophoresis at high pH. This results in structures resembling comets which by using suitable fluorescent stain, can be observed by fluorescent microscopy. Based on their size

  3. Is the Comet Assay a Sensitive Procedure for Detecting Genotoxicity?

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    Satomi Kawaguchi

    2010-01-01

    Full Text Available Although the Comet assay, a procedure for quantitating DNA damage in mammalian cells, is considered sensitive, it has never been ascertained that its sensitivity is higher than the sensitivity of other genotoxicity assays in mammalian cells. To determine whether the power of the Comet assay to detect a low level of genotoxic potential is superior to those of other genotoxicity assays in mammalian cells, we compared the results of Comet assay with those of micronucleus test (MN test. WTK1 human lymphoblastoid cells were exposed to methyl nitrosourea (MNU, ethyl nitrosourea (ENU, methyl methanesulfonate (MMS, ethyl methanesulfonate (EMS, bleomycin (BLM, or UVC. In Comet assay, cells were exposed to each mutagen with (Comet assay/araC and without (Comet assay DNA repair inhibitors (araC and hydroxyurea. Furthermore, acellular Comet assay (acellular assay was performed to determine how single-strand breaks (SSBs as the initial damage contributes to DNA migration and/or to micronucleus formation. The lowest genotoxic dose (LGD, which is defined as the lowest dose at which each mutagen causes a positive response on each genotoxicity assay, was used to compare the power of the Comet assay to detect a low level of genotoxic potential and that of MN test; that is, a low LGD indicates a high power. Results are summarized as follows: (1 for all mutagens studied, LGDs were MN test ≦ Comet assay; (2 except for BLM, LGDs were Comet assay/araC ≦ MN test; (3 except for UVC and MNU, LGDs were acellular assayComet assay/araC ≦ MN test ≦ Comet assay. The following is suggested by the present findings: (1 LGD in the Comet assay is higher than that in MN test, which suggests that the power of the MN test to detect a low level of genotoxic potential is superior to that of the Comet assay; (2 for the studied mutagens, all assays were able to detect all mutagens correctly, which suggests that the sensitivity of the Comet assay and that of the MN test were

  4. An ECVAG trial on assessment of oxidative damage to DNA measured by the comet assay

    DEFF Research Database (Denmark)

    Johansson, Clara; Møller, Peter; Forchhammer, Lykke

    2010-01-01

    The increasing use of single cell gel electrophoresis (the comet assay) highlights its popularity as a method for detecting DNA damage, including the use of enzymes for assessment of oxidatively damaged DNA. However, comparison of DNA damage levels between laboratories can be difficult due to dif...

  5. Comet assay for rapid detection of base damage in foods

    International Nuclear Information System (INIS)

    Al-Zubaidi, I. A.; Abdullah, T. S.; Qasim, S. R.

    2012-12-01

    Single cell gel electrophoresis (SCGE) or comet assay technique a sensitive, reliable and rapid method for DNA double and single strand break, alkali- labile site and delayed repair site detection in individual cells. In recent years, this method has been widely used for studies of DNA repair, genetic toxicology, and environmental biomontoring, however, this technique serves as an important tool for detection of DNA damage in living organism and is increasing being used in genetic testing of industrial chemicals, environmental agent's contaminations. This research paper helps to evaluate the oxidant agent's effects of exposure to organic pollutants by using comet assay techniques. This study used five samples of each food sample (Meat, Chicken, Rice, Fruits, Vegetables and Tea) to evaluate the genotoxic effects of exposure, to environmental agent's pollutants. The experimental data suggest that the DNA damage parameters ( Tail length, Tail width 1 ) were found higher value in exposed population when compared with the ratio of the length to width that cells exhibiting no migration having a ratio of 1. The percentage and distribution of cells in exposed population of cells also increases with the increase in values. This study demonstrates that, using sensitive techniques, it is possible to detect environmental agent's risks at an early stage. (Author)

  6. Micropatterned comet assay enables high throughput and sensitive DNA damage quantification

    Science.gov (United States)

    Ge, Jing; Chow, Danielle N.; Fessler, Jessica L.; Weingeist, David M.; Wood, David K.; Engelward, Bevin P.

    2015-01-01

    The single cell gel electrophoresis assay, also known as the comet assay, is a versatile method for measuring many classes of DNA damage, including base damage, abasic sites, single strand breaks and double strand breaks. However, limited throughput and difficulties with reproducibility have limited its utility, particularly for clinical and epidemiological studies. To address these limitations, we created a microarray comet assay. The use of a micrometer scale array of cells increases the number of analysable comets per square centimetre and enables automated imaging and analysis. In addition, the platform is compatible with standard 24- and 96-well plate formats. Here, we have assessed the consistency and sensitivity of the microarray comet assay. We showed that the linear detection range for H2O2-induced DNA damage in human lymphoblastoid cells is between 30 and 100 μM, and that within this range, inter-sample coefficient of variance was between 5 and 10%. Importantly, only 20 comets were required to detect a statistically significant induction of DNA damage for doses within the linear range. We also evaluated sample-to-sample and experiment-to-experiment variation and found that for both conditions, the coefficient of variation was lower than what has been reported for the traditional comet assay. Finally, we also show that the assay can be performed using a 4× objective (rather than the standard 10× objective for the traditional assay). This adjustment combined with the microarray format makes it possible to capture more than 50 analysable comets in a single image, which can then be automatically analysed using in-house software. Overall, throughput is increased more than 100-fold compared to the traditional assay. Together, the results presented here demonstrate key advances in comet assay technology that improve the throughput, sensitivity, and robustness, thus enabling larger scale clinical and epidemiological studies. PMID:25527723

  7. OpenComet: An automated tool for comet assay image analysis

    Directory of Open Access Journals (Sweden)

    Benjamin M. Gyori

    2014-01-01

    Full Text Available Reactive species such as free radicals are constantly generated in vivo and DNA is the most important target of oxidative stress. Oxidative DNA damage is used as a predictive biomarker to monitor the risk of development of many diseases. The comet assay is widely used for measuring oxidative DNA damage at a single cell level. The analysis of comet assay output images, however, poses considerable challenges. Commercial software is costly and restrictive, while free software generally requires laborious manual tagging of cells. This paper presents OpenComet, an open-source software tool providing automated analysis of comet assay images. It uses a novel and robust method for finding comets based on geometric shape attributes and segmenting the comet heads through image intensity profile analysis. Due to automation, OpenComet is more accurate, less prone to human bias, and faster than manual analysis. A live analysis functionality also allows users to analyze images captured directly from a microscope. We have validated OpenComet on both alkaline and neutral comet assay images as well as sample images from existing software packages. Our results show that OpenComet achieves high accuracy with significantly reduced analysis time.

  8. Correlation between the genotoxicity endpoints measured by two different genotoxicity assays: comet assay and CBMN assay

    Directory of Open Access Journals (Sweden)

    Carina Ladeira

    2015-06-01

    The results concerning of positive findings by micronuclei and non significant ones by comet assay, are corroborated by Deng et al. (2005 study performed in workers occupationally exposed to methotrexate, also a cytostatic drug. According to Cavallo et al. (2009, the comet assay seems to be more suitable for the prompt evaluation of the genotoxic effects, for instance, of polycyclic aromatic hydrocarbons mixtures containing volatile substances, whereas the micronucleus test seems more appropriate to evaluate the effects of exposure to antineoplastic agents. However, there are studies that observed an increase in both the comet assay and the micronucleus test in nurses handling antineoplastic drugs, although statistical significance was only seen in the comet assay, quite the opposite of our results (Maluf & Erdtmann, 2000; Laffon et al. 2005.

  9. Assessment of Genotoxicity of Ionizing radiation using Tradescantia-Comet assay

    Energy Technology Data Exchange (ETDEWEB)

    Han, Min; Ryu, Tae Ho; Hyun, Kyung Man; Kim, Jin Kyu [Korea Atomic Energy Research Institute, Jeongeup (Korea, Republic of); Wilhelmova, Nad [Institute of Experimental Botany, Prague (Czech Republic)

    2010-05-15

    Over the last two decades, several new methodologies for the detection of DNA damage have been developed. The comet assay is currently used in different areas of biological sciences to detect DNA damage. The comet assay, also called the single cell gel electrophoresis (SCGE) was first introduced by Ostling and Johanson as a microelectrophoretic technique for the direct visualization of DNA damage in individual cells. The comet assay, due to its simplicity, sensitivity and need of a few cells, is ideal as a short-term genotoxicity test. The comet assay can theoretically be applied to every type of eukaryotic cell, including plant cells. Plants are very useful as monitors of genetic effects caused by pollution in the atmosphere, water and soil. Although the genotoxic effects detected by Tradescantia tests cannot be associated with mutagenesis or even carcinogenesis in humans, these bioassays are very useful tools for screening the mutagenic potential in the environment. Experiments were conducted to study the genotoxic effects of ionizing radiations on the genome integrity, particularly of Tradescantia. The increasingly frequent use of Tradescantia as a sensitive environmental bioindicator of genotoxic effects. This study was designed to assess the genotoxicity of ionizing radiation using Tradescnatia-comet assay

  10. Application of the comet assay in studies of programmed cell death (PCD in plants

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    Maria Charzyńska

    2014-01-01

    Full Text Available Programmed cell death (PCD in plants is an intensively investigated process. One of the main characteristics of PCD in both animal and plant organisms is the non-random, internucleosomal fragmentation of nuclear DNA, usually analysed using total DNA gel electrophoresis or TUNEL method. In this paper we present application of the "comet assay" (Single Cell Gel Electrophoresis for detection of nDNA degradation in studies of PCD during plant life cycle. We analyzed three types of tissue: anther tapetum, endosperm and mesophyll which were prepared in different ways to obtain a suspension of viable cells (without cell walls. The comet assay gives a possibility of examination of the nDNA degradation in individual cell. This method is significant for studies of the plant tissue differentiation and senescence especially in the cases when it is not possible to isolate large number of cells at the same developmental stage.

  11. Application of DNA comet assay for detection of radiation treatment of grams and pulses.

    Science.gov (United States)

    Khan, Hasan M; Khan, Ashfaq A; Khan, Sanaullah

    2011-12-01

    Several types of whole pulses (green lentils, red lentils, yellow lentils, chickpeas, green peas, cowpeas and yellow peas) and grams (black grams, red grams and white grams) have been investigated for the identification of radiation treatment using microgel electrophoresis of single cells (DNA comet assay). Pulses and grams were exposed to the radiation doses of 0.5, 1.0 and 5 kGy covering the legalized commercial dose range for protection from insect/pest infestations. All irradiated samples showed comet like stretching of fragmented DNA toward anode, which is expected for irradiated samples. Unirradiated samples showed many intact cells/nuclei in form of round stains or with short faint tails, which is typical for unirradiated food samples. The study shows that DNA comet assay can be used as a rapid, inexpensive and highly effective screening test for the detection of radiation treatment of foods, like pulses and grams.

  12. Interpreting sperm DNA damage in a diverse range of mammalian sperm by means of the two-tailed comet assay

    Directory of Open Access Journals (Sweden)

    Elva I eCortes-Gutierrez

    2014-11-01

    Full Text Available Sperm DNA damage is associated with fertilization failure, impaired pre-and post- embryo implantation and poor pregnancy outcome. A series of methodologies to assess DNA damage in spermatozoa have been developed but most are unable to differentiate between single-stranded DNA breaks (SSBs and double-stranded DNA breaks (DSBs on the same sperm cell. The two-dimensional Two-Tailed Comet assay (TT-comet protocol highlighted in this review overcomes this limitation and emphasizes the importance in accounting for difference in sperm protamine composition at a species-specific level for the appropriate preparation of the assay. The TT-comet is a modification of the original comet assay that uses a two dimensional electrophoresis to allow for the simultaneous evaluation of relatively high numbers of DSBs and SSBs in mammalian spermatozoa. Here we have compiled a retrospective overview of how the TT-comet assay has been used to investigate the structure and function of sperm DNA across a diverse range of mammalian species (eutheria, metatheria and prototheria. When conducted as part of the TT-comet assay, we discuss how (a the alkaline comet single assay has been used to help understand the constitutive and transient changes in DNA structure associated with chromatin packing, (b the capacity of the TT-comet to differentiate between the presence of SSBs and DSBs (c and the possible implications of SSBs or DSBs for the assessment of infertility.

  13. 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

  14. Reasonable threshold value used to segment the individual comet from the comet assay image

    International Nuclear Information System (INIS)

    Yan Xuekun; Chen Ying; Du Jie; Zhang Xueqing; Luo Yisheng

    2009-01-01

    Reasonable segmentation of the individual comet contour from the Comet Assay (CA) images is the precondition for all of parameters analysis during the automatic analyzing for the CA. The Otsu method and several arithmetic operators for image segmentation, such as Sobel, Prewitt, Roberts and Canny were used to segment the comet contour, and characters of the CA images were analyzed firstly. And then the segmentation methods which had been adopted in the software for CA automatic analysis, such as the CASP, the TriTek CometScore TM , were put for-ward and compared. At last, a two-step procedure for threshold calculation based on image-content analysis is adopted to segment the individual comet from the CA images, and several principles for the segmentation are put forward too.(authors)

  15. The use of comet assay in plant toxicology: recent advances

    Directory of Open Access Journals (Sweden)

    Conceição LV Santos

    2015-06-01

    Full Text Available The systematic study of genotoxicity in plants induced by contaminants and other stress agents has been hindered to date by the lack of reliable and robust biomarkers. The comet assay is a versatile and sensitive method for the evaluation of DNA damages and DNA repair capacity at single-cell level. Due to its simplicity and sensitivity, and the small number of cells required to obtain robust results, the use of plant comet assay has drastically increased in the last decade. For years its use was restricted to a few model species, e.g. Allium cepa, Nicotiana tabacum, Vicia faba, or Arabidopsis thaliana but this number largely increased in the last years. Plant comet assay has been used to study the genotoxic impact of radiation, chemicals including pesticides, phytocompounds, heavy metals, nanoparticles or contaminated complex matrices. Here we will review the most recent data on the use of this technique as a standard approach for studying the genotoxic effects of different stress conditions on plants. Also, we will discuss the integration of information provided by the comet assay with other DNA-damage indicators, and with cellular responses including oxidative stress, cell division or cell death. Finally, we will focus on putative relations between transcripts related with DNA damage pathways, DNA replication and repair, oxidative stress and cell cycle progression that have been identified in plant cells with comet assays demonstrating DNA damage.

  16. The comet assay – from toy to tool

    Directory of Open Access Journals (Sweden)

    Guenter Speit

    2015-04-01

    Full Text Available The comet assay is nowadays the most common method for measuring DNA damage and repair in single cells. It is based on the microelectrophoretic study published by Ostling and Johanson (1984 and was developed by Singh and coworkers (1988 to a versatile technique for quantitation of low levels of DNA damage in individual cells. This alkaline version still is the basis for the triumphant success of the comet assay in basic research into mechanisms of DNA damage and DNA repair, genotoxicity testing, ecotoxicology and human biomonitoring. Important technical improvements (e.g., the use of precoated slides, introduction of image analysis, high throughput methods, automated scoring systems made the assay more robust and more efficient. Modifications of the standard protocol provide more specific information on the type and biological significance of the damage studied. The introduction of lesion-specific endonucleases allowed the characterization of oxidative base damage, alkylation damage and UV-induced pyrimidine dimers. The combination with fluorescence in situ hybridization (FISH made it possible to identify DNA of particular chromosome regions and measure effects of damage and repair in particular genes. The comet assay is being increasingly used in genotoxicity testing. In particular, the in vivo comet assay has become a component of some genotoxicity test strategies and generally accepted test protocols have evolved over the years. A large international collaborative trial sponsored by the Japanese Center for the Validation of Alternative Methods (JaCVAM was recently completed and an OECD test guideline was approved. The comet assay is widely used in human biomonitoring to measure DNA damage as a marker of exposure to genotoxic agents or to investigate genoprotective effects. However, there are still problems in comparing results from different laboratories and there is need for reducing inter-laboratory variation and identification of standard

  17. Alkaline gel electrophoresis assay to detect DNA strand breaks and repair mechanisms in Escherichia coli

    International Nuclear Information System (INIS)

    Mattos, Jose Carlos Pelielo de; Motta, Ellen Serri da; Oliveira, Marcia Betania Nunes de; Dantas, Flavio Jose da Silva; Araujo, Adriano Caldeira de

    2008-01-01

    Reactive oxygen species (ROS) can induce lesions in different cellular targets, including DNA. Stannous chloride (SnCl 2 ) is a ROS generator, leading to lethality in Escherichia coli (E. coli), with the base excision repair (BER) mechanism playing a role in this process. Many techniques have been developed to detect genotoxicity, as comet assay, in eukaryotic cells, and plasmid DNA agarose gel electrophoresis. In this study, an adaptation of the alkaline gel electrophoresis method was carried out to ascertain the induction of strand breaks by SnCl 2 in bacterial DNA, from E. coli BER mutants, and its repair pathway. Results obtained show that SnCl 2 was able to induce DNA strand breaks in all strains tested. Moreover, endonuclease IV and exonuclease III play a role in DNA repair. On the whole, data has shown that the alkaline gel electrophoresis assay could be used both for studying DNA strand breaks induction and for associated repair mechanisms. (author)

  18. Ecotoxicological Assessment of Aquatic Genotoxicity Using the Comet Assay

    Directory of Open Access Journals (Sweden)

    KHUSNUL YAQIN

    2006-09-01

    Full Text Available Comet assay is a novel biological analysis, which is a sensitive, flexible, simple, rapid, and inexpensive method to assess aquatic genotoxicant. Since Singh and co-workers developed the method in 1988, its use has increased exponentially in various fields. This review discourses on the application of this assay in aquatic ecosystems. Various types of cells from various aquatic organisms have been tested by various genotoxicant both direct- and indirect-acting using the comet assay. The applications of this assay suggest that it is a useful assay to assess aquatic genotoxicants. However, there are some factors, which should be taken into account when using this assay as aquatic ecotoxicological assessment device such as inter-animal and cell variability.

  19. Application of the DNA ''comet assay'' to detect irradiation treatment of foods

    International Nuclear Information System (INIS)

    Delincee, H.

    1996-01-01

    Since treatment with ionising radiation causes DNA fragmentation, microgel electrophoresis of single cells (''comet assay'') offers a simple and rapid tool for identification of irradiated foods. The principle is based on migration of DNA in an agarose gel exposed to an electric field. Single cells or nuclei are embedded in the agarose and after lysis, intact DNA will virtually not move out of the cell upon electrophoresis, whereas if DNA has been fragmented, the fragments are able to migrate and ''comets'' following the cells become visible after staining. The advantages of this test, however, for foods not exposed to heat are its speed and simplicity, as the electrophoretic separation only requires a few minutes. DNA is visualised by silver staining, avoiding the need for a fluorescence microscope. Thus the method requires only relatively cheap equipment - in contrast to other methods for identification of irradiated food such as electron spin resonance (ESR) spectroscopy or gas chromatography/mass spectrometry (GC/MS). The DNA ''comet assay'' , therefore, seems suitable as a pre-screening test to detect whether food has been radiation processed. Suspected samples may subsequently be analysed by established, but more expensive techniques. (author)

  20. CometQ: An automated tool for the detection and quantification of DNA damage using comet assay image analysis.

    Science.gov (United States)

    Ganapathy, Sreelatha; Muraleedharan, Aparna; Sathidevi, Puthumangalathu Savithri; Chand, Parkash; Rajkumar, Ravi Philip

    2016-09-01

    DNA damage analysis plays an important role in determining the approaches for treatment and prevention of various diseases like cancer, schizophrenia and other heritable diseases. Comet assay is a sensitive and versatile method for DNA damage analysis. The main objective of this work is to implement a fully automated tool for the detection and quantification of DNA damage by analysing comet assay images. The comet assay image analysis consists of four stages: (1) classifier (2) comet segmentation (3) comet partitioning and (4) comet quantification. Main features of the proposed software are the design and development of four comet segmentation methods, and the automatic routing of the input comet assay image to the most suitable one among these methods depending on the type of the image (silver stained or fluorescent stained) as well as the level of DNA damage (heavily damaged or lightly/moderately damaged). A classifier stage, based on support vector machine (SVM) is designed and implemented at the front end, to categorise the input image into one of the above four groups to ensure proper routing. Comet segmentation is followed by comet partitioning which is implemented using a novel technique coined as modified fuzzy clustering. Comet parameters are calculated in the comet quantification stage and are saved in an excel file. Our dataset consists of 600 silver stained images obtained from 40 Schizophrenia patients with different levels of severity, admitted to a tertiary hospital in South India and 56 fluorescent stained images obtained from different internet sources. The performance of "CometQ", the proposed standalone application for automated analysis of comet assay images, is evaluated by a clinical expert and is also compared with that of a most recent and related software-OpenComet. CometQ gave 90.26% positive predictive value (PPV) and 93.34% sensitivity which are much higher than those of OpenComet, especially in the case of silver stained images. The

  1. Benzene-induced genotoxicity in mice in vivo detected by the alkaline comet assay

    DEFF Research Database (Denmark)

    Tuo, J; Loft, S; Thomsen, M S

    1996-01-01

    The myelotoxic and genotoxic effects of benzene have been related to oxidative DNA damage after metabolism by CYP2E1. Single cell gel electrophoresis (alkaline comet assay) detects DNA damage and may thus be a convenient method for the study of benzene genotoxicity. Benzene exposure to NMRI mice...... as a single oral gavage at 40, 200 or 450 mg/kg resulted in dose-related DNA damage indicated by an increased comet tail length of peripheral blood lymphocytes and bone marrow nucleated cells sampled 6 h after exposure. After a dose of 40 mg/kg, there was a 1.6-fold increase of 'tail length' in bone marrow.......01). By comparing our data with those from genotoxicity studies on benzene using other methods, we conclude that the 'alkaline comet assay' is a sensitive method to detect DNA damage induced by benzene. We also infer that CYP2E1 contributes, at least partly, to the formation of the 'comet'-inducing metabolites...

  2. Reference cells and ploidy in the comet assay

    Directory of Open Access Journals (Sweden)

    Gunnar eBrunborg

    2015-02-01

    Full Text Available In the comet assay, single cells are analyzed with respect to their level of DNA damage. Discrimination of the individual cell or cell type based on DNA content, with concomitant scoring of the DNA damage, is useful since this may allow analysis of mixtures of cells. Different cells can then be characterized based on their ploidy, cell cycle stage, or genome size. We here describe two applications of such a cell type-specific comet assay: (i Testicular cell suspensions, analyzed on the basis of their ploidy during spermatogenesis; and (ii reference cells in the form of fish erythrocytes which can be included as internal standards to correct for inter-assay variations. With standard fluorochromes used in the comet assay, the total staining signal from each cell – whether damaged or undamaged – was found to be associated with the cell’s DNA content. Analysis of the fluorescence intensity of single cells is straightforward since these data are available in scoring systems based on image analysis. The analysis of testicular cell suspensions provides information on cell type specific composition, susceptibility to genotoxicants, and DNA repair. Internal reference cells, either untreated or carrying defined numbers of lesions induced by ionizing radiation, are useful for investigation of experimental factors that can cause variation in comet assay results, and for routine inclusion in experiments to facilitate standardization of methods and comparison of comet assay data obtained in different experiments or in different laboratories. They can also be used - in combination with a reference curve - to quantify the DNA lesions induced by a certain treatment. Fish cells of a range of genome sizes, both greater and smaller than human, are suitable for this purpose and they are inexpensive.

  3. The comet assay in testing the potential genotoxicity of nanomaterials

    Directory of Open Access Journals (Sweden)

    Amaya Azqueta

    2015-06-01

    Full Text Available In the last two decades the production and use of nanomaterials (NMs has impressively increased. Their small size, given a mass equal to that of the corresponding bulk material, implies an increase in the surface area and consequently in the number of atoms that can be reactive. They possess different physical, chemical and biological properties compared to bulk materials of the same composition, which makes them very interesting and valuable for many different applications in technology, energy, construction, electronics, agriculture, optics, paints, textiles, food, cosmetics, medicine... Toxicological assessment of NMs is crucial; the same properties that make them interesting also make them potentially harmful for health and the environment. However, the term NM covers many different kinds of particle , and so there is no simple, standard approach to assessing their toxicity. NMs can enter the cell, interact with cell components and even penetrate the nucleus and interfere with the genetic material. Among the different branches of toxicology, genotoxicity is a main area of concern since it is closely related with the carcinogenic potential of compounds. The Organisation for Economic Co-operation and Development (OECD has published internationally agreed in vitro and in vivo validated test methods to evaluate different genotoxic endpoints of chemicals, including chromosome and gene mutations, and DNA breaks. However not all the assays are suitable to study the genotoxic potential of NMs as has been shown by the OECD Working Party on Manufactured Nanomaterials (WPMN. Moreover, alterations to DNA bases, which are precursors to mutations and of great importance in elucidating the mechanism of action of NMs, are not covered by the OECD guidelines. The in vivo standard comet assay (which measures DNA breaks and alkali-labile sites was included in the OECD assays battery in September 2014 while the in vitro standard comet assay is currently under

  4. Variation in assessment of oxidatively damaged DNA in mononuclear blood cells by the comet assay with visual scoring

    DEFF Research Database (Denmark)

    Forchhammer, Lykke; Bräuner, Elvira Vaclavik; Folkmann, Janne Kjaersgaard

    2008-01-01

    The comet assay is popular for assessments of genotoxicity, but the comparison of results between studies is challenging because of differences in experimental procedures and reports of DNA damage in different units. We investigated the variation of DNA damage in mononuclear blood cells (MNBCs......) measured by the comet assay with focus on the variation related to alkaline unwinding and electrophoresis time, number of cells scored, as well as the putative benefits of transforming the primary end points to common units by the use of reference standards and calibration curves. Eight experienced......, our results indicate that inter-investigator difference in scoring is a strong determinant of DNA damage levels measured by the comet assay....

  5. Protective effect of peach kernel extracts on radiation-induced DNA damage in human blood lymphocytes in the comet assay

    International Nuclear Information System (INIS)

    Kim, Jin Kyu; Lee, Chang Joo; Park, Tae Won; Chai, Young Gyu

    1999-01-01

    The alkaline single-cell gel electrophoresis (SCGE) assay, the comet assay, has been applied to the detection of DNA damage from a number of chemical and biological factors in vivo and in vitro. The comet assay is a novel method to assess DNA single-strand breaks, alkali-labile sites in individual cells. We evaluated the effect of peach kernel extracts on radiation-induced DNA damage in human blood lymphocytes using the comet assay. The lymphocytes, with or without pretreatment of the extracts, were exposed to 0, 0.1, 0.3, 0.5, 1.0 and 2.0 Gy of 60 Co gamma ray. Significantly increased tail moment, which was a marker of DNA strand breaks in the comet assay, showed an excellent dose-response relationship. The treatment of the peach kernel extracts prominently reduced the DNA damage in irradiated groups compared to that in non-treated control groups. The result indicated that the extracts showed radioprotective effect on lymphocyte DNA when assessed by the comet assay

  6. Application of the Single Cell Gel Electrophoresis (SCGE) Assay to Genotoxicity Evaluation in Plants and Animals

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Jin Kyu

    2007-10-15

    Application of the Single Cell Gel Electrophoresis (SCGE) Assay to Genotoxicity Evaluation in Plants and Animals. Recently, the importance of ionizing radiation and chemicals has been recognized since radio- and chemical therapy is directly related to the control of various diseases such as cancer. Radiation and the chemicals can cause biological damages while they have great applicability. It is of necessity to analyze rapidly, easily and accurately the biological effects, especially DNA damage due to those factors. Recently SCGE (single cell gel electrophoresis assay, alias comet assay) has been developed for the efficient evaluation of DNA damage. In this report, the comprehensive review will be given on the rationale, the technical applications and the advantages and shortcomings of SCGE assay. This method can be directly applied to study on toxicity, cancer, and aging in terms of the evaluation of DNA damages due to radiation and chemicals on human cellular level. It is also suggested that comet assay be used for testing genotoxicity of suspected substances, detecting irradiated foods, screening radioprotective candidates, and studying DNA repair process in various biological systems.

  7. Detection of garlic gamma-irradiated by assay comet

    Energy Technology Data Exchange (ETDEWEB)

    Moreno Alvarez, Damaris L.; Miranda, Enrique F. Prieto; Carro, Sandra; Iglesias Enrique, Isora; Matos, Wilberto [Centro de Aplicaciones Tecnologicas y Desarrollo Nuclear (CEADEN), Ciudad de La Habana (Cuba)], e-mail: damaris@ceaden.edu.cu

    2009-07-01

    The garlic samples were irradiated in a facility with {sup 60}Co sources, at absorbed dose values of 0-0,15 kGy. The detection method utilized for the identification of the irradiated garlic was biological comet assay. The samples were classified post-irradiation several times. The irradiated samples showed high strand breaks of DNA exhibiting comets of several forms, while the not irradiated and lower dose samples showed a behavior like round shape and light comets. Significant differences were found for higher absorbed dose values at 0.06 kGy, this absorbed dose value is corresponding with the applied dose value at this food in order to avoid the germination. (author)

  8. Detection of garlic gamma-irradiated by assay comet

    International Nuclear Information System (INIS)

    Moreno Alvarez, Damaris L.; Miranda, Enrique F. Prieto; Carro, Sandra; Iglesias Enrique, Isora; Matos, Wilberto

    2009-01-01

    The garlic samples were irradiated in a facility with 60 Co sources, at absorbed dose values of 0-0,15 kGy. The detection method utilized for the identification of the irradiated garlic was biological comet assay. The samples were classified post-irradiation several times. The irradiated samples showed high strand breaks of DNA exhibiting comets of several forms, while the not irradiated and lower dose samples showed a behavior like round shape and light comets. Significant differences were found for higher absorbed dose values at 0.06 kGy, this absorbed dose value is corresponding with the applied dose value at this food in order to avoid the germination. (author)

  9. Comet assay as a cold chain control tool

    International Nuclear Information System (INIS)

    Duarte, Renato Cesar

    2009-01-01

    Bearing in mind an ever more demanding market regarding the quality of food, it has been necessary to develop processes that meet the demands of consumers. Within the existing processes the cold chain and irradiation stand out. The cold chain comprises all the stages of conserving food from production, cooling, freezing, storing and transportation to the final consumer. Irradiation, as a means of conserving food, prolongs the shelf life, inhibits budding and reduces pathogenic contamination among other benefits. Is very important the identification of food degradation in function of failure on the processes which they were subjected. The comet assay is a screening test widely studied, considerate fast and with low cost. By the fact of the test identify breaks on the DNA, may be possible use the comet test on the control of cold chain failures that degrade de food. The labels and stamp, do not consider the previous food situation and indicate failures from the moment where they be placed in contact with the product. With the comet assay is possible to check the degradation that has occurred in liver chicken samples until the moment of comet's test realization. (author)

  10. Optimization of Neutral Comet Assay for studying DNA double-strand breaks in pea and wheat

    Directory of Open Access Journals (Sweden)

    Ivelina Nikolova

    2013-01-01

    Full Text Available This study describes an adaptation of the Comet assay under neutral conditions for mono- and dicotyledonous plants pea (Pisum sativum L. and wheat (Triticum aestivum L.. Modifications concern lysis and electrophoresis steps, respectively. Electrophoresis was carried out varying the intensity of the electric field. A linear relationship between the percentages of DNA in the tail from control background with alteration of intensity was found. Trypan blue dye exclusion test was used in order to determine the intactness of nuclear membrane of the isolated nuclei from both plant model systems. Assessment was conducted on non-irradiated and irradiated nuclei on a monolayer with three doses of UVC. It was found that the share of intact nuclei (trypan blue negative ones is about 95% in controls. Gradual dose-related increase of damaged nuclei was observed in both species, reaching statistical significance only at the higher dose applied.

  11. Assessment of radiation induced cytogenetic damage in human keratinocytes by comet assay

    International Nuclear Information System (INIS)

    Joseph, Praveen; Sanjeev Ganesh; Narayana, Y.; Puthali, Abhay; Bhat, N.N.

    2010-01-01

    In the present study the effect of gamma radiation on normal human keratinocytes (HaCaT) cells has been analyzed using alkaline comet assay and a comparative study over the sensitivity of different comet parameters such as tail length (TL), olive tail moment (OTM) and percentage tail DNA (TDNA) has also been made. Human keratinocytes (HaCaT) cells were grown in Dulbecco's modified essential medium (DMEM) (10% FCS) at 37 °C in a humidified atmosphere containing 5% CO 2 . Cultured cells were harvested with 0.025 % trypsin EDTA. The sample (2 X 10 cells/ml) was exposed to gamma radiation of different dose using a 60 Co gamma source at dose rate of 2 Gy min -1 and the dosimetry has been carried out using Fricke and FBX dosimeters. After irradiation, to quantify the DNA damage the comet assay (single cell gel electrophoresis) was carried out under alkaline conditions, by the methods outlined by Singh et al. The quantification of the DNA strand breaks in each cells were performed using CASP software. The DNA damage quantification can be accomplished by measuring those comet parameters which exhibit a linear dependence on the amount of DNA damage. In the present study, comet parameters such as OTM, TL and TDNA were recorded and the variation of these parameters and their correlation coefficients for different doses of gamma radiation is plotted. The OTM value is normalized with control value and control for TL and TDNA is adjusted to zero to avoid initial variations in different experiments

  12. Indentification of radiation treatment of frozen chicken and fresh turkey using DNA comet assay

    International Nuclear Information System (INIS)

    Khan, H.M.; Khan, A.A.

    2009-01-01

    DNA comet assay was applied to check the detection of radiation treatment of frozen chicken and fresh turkey. The cells from unirradiated and irradiated samples of chicken and turkey were extracted in cold PBS, embedded in agarose on microscope slides, lysed for 15 minutes in 2.5 % SDS and subjected to electrophoresis at a rate of 2V/cm for 2 minutes. After silver staining these slides were evaluated through an ordinary transmission microscope. In irradiated samples, fragmented DNA (due to radiation treatment) stretched towards the anode and cells appear as comets. The density of DNA material in the tails increased with increasing radiation dose. However in case of unirradiated samples, the large molecule of DNA remained relatively intact and there was only minor or no migration of DNA; thus cells were round or had very short tails. Therefore, clear discrimination between unirradiated and irradiated food samples is possible. Hence, DNA comet assay provides an inexpensive and quick screening method for several kinds of foods containing DNA including frozen chicken and fresh turkey. (author)

  13. Bovine Papillomavirus Clastogenic Effect Analyzed in Comet Assay

    Science.gov (United States)

    Araldi, R. P.; Melo, T. C.; Diniz, N.; Mazzuchelli-de-Souza, J.; Carvalho, R. F.; Beçak, W.; Stocco, R. C.

    2013-01-01

    Bovine papillomavirus (BPV) is an oncogenic virus related to serious livestock diseases. Oncoproteins encoded by BPV are involved in several steps of cellular transformation and have been reported as presenting clastogenic effects in peripheral lymphocytes and primary culture cells. The aim of this study was to evaluate the clastogenic potential of BPV types 1, 2, and 4 by comet assay. Peripheral blood was collected from 37 bovines, 32 infected with different levels of papillomatosis (12 animals have no affection) and five calves, virus free (negative control). The viral identification showed presence of more than one virus type in 59.375% of the infected animals. Comet assay was performed according to alkaline technique. The Kruskal-Wallis test showed statistical difference between the negative control group and infected animals (P = 0.0015). The Dunn post hoc test showed difference comparing the infected animals with calves. Mann-Whitney U test verified no difference between animals infected with only one viral type and animals presenting more than one viral type. The comet assay is considered an efficient tool for assessment of damage in the host chromatin due to viral action, specifically highlighting viral activity in blood cells. PMID:23956996

  14. Bovine Papillomavirus Clastogenic Effect Analyzed in Comet Assay

    Directory of Open Access Journals (Sweden)

    R. P. Araldi

    2013-01-01

    Full Text Available Bovine papillomavirus (BPV is an oncogenic virus related to serious livestock diseases. Oncoproteins encoded by BPV are involved in several steps of cellular transformation and have been reported as presenting clastogenic effects in peripheral lymphocytes and primary culture cells. The aim of this study was to evaluate the clastogenic potential of BPV types 1, 2, and 4 by comet assay. Peripheral blood was collected from 37 bovines, 32 infected with different levels of papillomatosis (12 animals have no affection and five calves, virus free (negative control. The viral identification showed presence of more than one virus type in 59.375% of the infected animals. Comet assay was performed according to alkaline technique. The Kruskal-Wallis test showed statistical difference between the negative control group and infected animals (P=0.0015. The Dunn post hoc test showed difference comparing the infected animals with calves. Mann-Whitney U test verified no difference between animals infected with only one viral type and animals presenting more than one viral type. The comet assay is considered an efficient tool for assessment of damage in the host chromatin due to viral action, specifically highlighting viral activity in blood cells.

  15. Bovine papillomavirus clastogenic effect analyzed in comet assay.

    Science.gov (United States)

    Araldi, R P; Melo, T C; Diniz, N; Mazzuchelli-de-Souza, J; Carvalho, R F; Beçak, W; Stocco, R C

    2013-01-01

    Bovine papillomavirus (BPV) is an oncogenic virus related to serious livestock diseases. Oncoproteins encoded by BPV are involved in several steps of cellular transformation and have been reported as presenting clastogenic effects in peripheral lymphocytes and primary culture cells. The aim of this study was to evaluate the clastogenic potential of BPV types 1, 2, and 4 by comet assay. Peripheral blood was collected from 37 bovines, 32 infected with different levels of papillomatosis (12 animals have no affection) and five calves, virus free (negative control). The viral identification showed presence of more than one virus type in 59.375% of the infected animals. Comet assay was performed according to alkaline technique. The Kruskal-Wallis test showed statistical difference between the negative control group and infected animals (P = 0.0015). The Dunn post hoc test showed difference comparing the infected animals with calves. Mann-Whitney U test verified no difference between animals infected with only one viral type and animals presenting more than one viral type. The comet assay is considered an efficient tool for assessment of damage in the host chromatin due to viral action, specifically highlighting viral activity in blood cells.

  16. Evaluation of environmental genotoxicity by comet assay in Columba livia.

    Science.gov (United States)

    González-Acevedo, Anahi; García-Salas, Juan A; Gosálvez, Jaime; Fernández, José Luis; Dávila-Rodríguez, Martha I; Cerda-Flores, Ricardo M; Méndez-López, Luis F; Cortés-Gutiérrez, Elva I

    2016-01-01

    The concentrations of recognized or suspected genotoxic and carcinogenic agents found in the air of large cities and, in particular, developing countries, have raised concerns about the potential for chronic health effects in the populations exposed to them. The biomonitoring of environmental genotoxicity requires the selection of representative organisms as "sentinels," as well as the development of suitable and sensitive assays, such as those aimed at assessing DNA damage. The aim of this study was to evaluate DNA damage levels in erythrocytes from Columba livia living in the metropolitan area of Monterrey, Mexico, compared with control animals via comet assay, and to confirm the results via Micronuclei test (MN) and DNA breakage detection-fluorescence in situ hybridization (DBD-FISH). Our results showed a significant increase in DNA migration in animals from the area assayed compared with that observed in control animals sampled in non-contaminated areas. These results were confirmed by MN test and DBD-FISH. In conclusion, these observations confirm that the examination of erythrocytes from Columba livia via alkaline comet assay provides a sensitive and reliable end point for the detection of environmental genotoxicants.

  17. Cytotoxicity and genotoxicity assessment of Euphorbia hirta in MCF-7 cell line model using comet assay

    OpenAIRE

    Kwan Yuet Ping; Ibrahim Darah; Yeng Chen; Sreenivasan Sasidharan

    2013-01-01

    Objective: To evaluate the cytotoxicity and genotoxicity activity of Euphorbia hirta (E. hirta) in MCF-7 cell line model using comet assay. Methods: The cytotoxicity of E. hirta extract was investigated by employing brine shrimp lethality assay and the genotoxicity of E. hirta was assessed by using Comet assay. Results: Both toxicity tests exhibited significant toxicity result. In the comet assay, the E. hirta extract exhibited genotoxicity effects against MCF-7 DNA in a time-dependent m...

  18. Detection of radiation treatment of frozen chicken using a quick microgel electrophoresis assay

    International Nuclear Information System (INIS)

    Khan, A.A.; Khan, H.M.

    2007-01-01

    DNA Comet Assay (microgel electrophoresis of single cell) is very useful and sensitive analytical technique to investigate several types of damages of DNA - especially the fragmentation. As DNA is the essential part of several foods, either animal or plant origin, therefore this method is versatile and valuable for detection of radiation treatment of several types of food. Ionizing radiation causes DNA breakage, fragmentation, base modification and single- or double-strand breaks. Therefore, the detection of fragmented DNA present in any kind or type of food can serve as the marker of detection of radiation treatment. The cells from the chicken samples (unirradiated and irradiated) were extracted in cold PBS, embedded in agarose on microscope slides, lysed for 15 minutes in 2.5 % SDS and electrophoressed at a rate of 2V /cm for 2 minutes. After silver staining, the slides were evaluated through an ordinary transmission microscope. In irradiated samples, fragmented DNA (due to radiation treatment) stretched towards the anode and cells appearing as comets. The density of DNA in the tails increased with increasing radiation dose. However, in non-irradiated samples, the large molecule of DNA remained relatively intact and there was only minor or no migration of DNA; the cells were round or had very short tails. Therefore, DNA comet assay provides an inexpensive, rapid and relatively simple screening method for several kinds of radiation-processed and untreated foods. (author)

  19. The single-cell gel electrophoresis assay to determine apoptosis ...

    African Journals Online (AJOL)

    The aim of the present study was to determine if the pattern of DNA fragmentation determined by the single cell gel electrophoresis assay can be used to determine apoptosis induced by siRNA in Colo 320 cells. When the frequency of appearance of apoptotic cells following was observed over a period of time, there was a ...

  20. Impaired DNA repair as assessed by the ``comet`` assay in patients developing thyroid carcinoma after radiotherapy

    Energy Technology Data Exchange (ETDEWEB)

    Leprat, F.; Sarasin, A.; Suarez, H.G. [Institut de Recherches sur le Cancer, 94 - Villesuif (France); Leprat, F.; Alapetite, C.; Rosselli, F.; Ridet, A.; Moustacch, E.; Alapetite, C. [Institut Curie, 75 - Paris (France); Schlumberger, M. [Institut Gustave Roussy, 94 - Villejuif (France)

    1997-03-01

    A defective cellular response to DNA lesions induced be genotoxic agents may be associated to an increased cancer proneness. This has been clearly identified in some rare but extensively studied genetic diseases such as xeroderma pigmentosum (XP), ataxia telangiectasia (AT) and Fanconi anemia (FA). In practical oncology, most patients receive genotoxic therapeutic agents and the presence of so far unidentified sensitive genotypes could account for an increased susceptibility to cancer in a subgroup of exposed patients. The thyroid gland of children is especially sensitive to the carcinogenic effect of ionizing radiation. Evidence for risk is reported even at doses as low as 0.1 Gy, and the excess relative risk to develop a thyroid tumor following a radiation dose of 1 Gy in childhood is of 7.7 [l]. In order to determine if a defect in repair of DNA strand breaks could be involved, as an early step, in the development of secondary thyroid tumors after radiotherapy, we examined, using the alkaline single cell gel electrophoresis assay (SCGE or `comet`), the response to in vitro {gamma}-rays exposure of lymphocytes of a small group of patients who developed thyroid carcinoma after radiotherapy for a primary tumor. Because of its practical advantages, the alkaline comet assay offers the opportunity to question the role of DNA strand beaks rejoining capacity of the individual in the radiation induced carcinogenesis of thyroid tumors. This preliminary study of a small group of patients with therapeutic irradiation at childhood for a primary tumor indicates that, at the time of blood sampling, lymphocytes of some of these patients demonstrated reduced rejoining capacity. These results suggest that the comet assay might help to distinguish a subgroup of individuals at risk for radiation induced genomic instability and encourage further investigation. (authors)

  1. Nonradioactive telomerase activity assay by microchip electrophoresis: privileges to the classical gel electrophoresis assay.

    Science.gov (United States)

    Zhelev, Zhivko; Bakalova, Rumiana; Ewis, Ashraf; Ohba, Hideki; Ishikawa, Mitsuru; Baba, Yoshinobu

    2005-08-01

    The present study accents on the privileges of microchip-based electrophoresis to the conventional gel electrophoresis in separation of telomerase repeat amplification protocol/polymerase chain reaction (PCR) ladder products obtained in telomerase-catalyzed reaction in cancer cells. We try to clarify the interpretation of the results obtained by both electrophoretic procedures and to avoid misinterpretation as a result of PCR-dependent artefacts.

  2. Reliability of plant root comet assay in comparison with human leukocyte comet assay for assessment environmental genotoxic agents.

    Science.gov (United States)

    Reis, Gabriela Barreto Dos; Andrade-Vieira, Larissa Fonseca; Moraes, Isabella de Campos; César, Pedro Henrique Souza; Marcussi, Silvana; Davide, Lisete Chamma

    2017-08-01

    Comet assay is an efficient test to detect genotoxic compounds based on observation of DNA damage. The aim of this work was to compare the results obtained from the comet assay in two different type of cells extracted from the root tips from Lactuca sativa L. and human blood. For this, Spent Pot Liner (SPL), and its components (aluminum and fluoride) were applied as toxic agents. SPL is a solid waste generated in industry from the aluminum mining and processing with known toxicity. Three concentrations of all tested solutions were applied and the damages observed were compared to negative and positive controls. It was observed an increase in the frequency of DNA damage for human leukocytes and plant cells, in all treatments. On human leukocytes, SPL induced the highest percentage of damage, with an average of 87.68%. For root tips cells of L. sativa the highest percentage of damage was detected for aluminum (93.89%). Considering the arbitrary units (AU), the average of nuclei with high levels of DNA fragmentation was significant for both cells type evaluated. The tested cells demonstrated equal effectiveness for detection of the genotoxicity induced by the SPL and its chemical components, aluminum and fluoride. Further, using a unique method, the comet assay, we proved that cells from root tips of Lactuca sativa represent a reliable model to detect DNA damage induced by genotoxic pollutants is in agreement of those observed in human leukocytes as model. So far, plant cells may be suggested as important system to assess the toxicological risk of environmental agents. Copyright © 2017 Elsevier Inc. All rights reserved.

  3. Identification of irradiated refrigerated pork with the DNA comet assay

    Energy Technology Data Exchange (ETDEWEB)

    Araujo, M.M. E-mail: villavic@net.ipen.br; Marin-Huachaca, N.S.; Mancini-Filho, J. E-mail: jmancini@usp.br; Delincee, H.; Villavicencio, A.L.C.H. E-mail: henry.delincee@bfe.uni-karlsruhe.de

    2004-10-01

    Food irradiation can contribute to a safer and more plentiful food supply by inactivating pathogens, eradicating pests and by extending shelf-life. Particularly in the case of pork meat, this process could be a useful way to inactivate harmful parasites such as Trichinella and Taenia solium. Ionizing radiation causes damage to the DNA of the cells (e.g. strand breaks), which can be used to detect irradiated food. Microelectrophoresis of single cells ('Comet Assay') is a simple and rapid test for DNA damage and can be used over a wide dose range and for a variety of products. Refrigerated pork meat was irradiated with a {sup 60}Co source, Gammacell 220 (A.E.C.L.) installed in IPEN (Sao Paulo, Brazil). The doses given were 0, 1.5, 3.0 and 4.5 kGy for refrigerated samples. Immediately after irradiation the samples were returned to the refrigerator (6 deg. C). Samples were kept in the refrigerator after irradiation. Pork meat was analyzed 1, 8 and 10 days after irradiation using the DNA 'Comet Assay'. This method showed to be an inexpensive and rapid technique for qualitative detection of irradiation treatment.

  4. Genotoxic effects of bismuth (III oxide nanoparticles by comet assay

    Directory of Open Access Journals (Sweden)

    Reecep Liman

    2015-06-01

    Full Text Available Bismuth oxide is one of the important transition metal oxides and it has been intensively studied due to their peculiar characteristics (semiconductor band gap, high refractive index, high dielectric permittivity, high oxygen conductivity, resistivity, photoconductivity and photoluminescence etc.. Therefore, it is used such as microelectronics, sensor technology, optical coatings, transparent ceramic glass manufacturing, nanoenergetic gas generator, biosensor for DNA hybridization, potential immobilizing platforms for glucose oxidase and polyphenol oxidase, fuel cells, a additive in paints, an astringent in a variety of medical creams and topical ointments, and for the determination of heavy metal ions in drinking water, mineral water and urine. In addition this, Bismuth (III oxide nanoparticles (BONPs are favorable for the biomolecules adsorption than regular sized particles because of their greater advantages and novel characteristics (much higher specific surface, greater surface free energy, and good electrochemical stability etc.. Genotoxic effects of BONPs were investigated on the root cells of Allium cepa by Comet assay. A. cepa roots were treated with the aqueous dispersions of BONPs at 5 different concentrations (12.5, 25, 50, 75, and 100 ppm for 4 h. A significant increase in DNA damage was also observed at all concentrations of BONPs except 12.5 ppm by Comet assay. The results were also analyzed statistically by using SPSS for Windows; Duncan’s multiple range test was performed. These result indicate that BONPs exhibit genotoxic activity in A. cepa root meristematic cells.

  5. Radio response of human lymphocytes pretreated with boron and gadoliniums assessed by the, comet assay

    International Nuclear Information System (INIS)

    Kim, J. K.; Park, T. W.; Cebulska-Wasiewska, A.; Nili, M.

    2009-01-01

    Boron and gadolinium are among the nuclides that hold a unique property of being a neutron capture therapy agent. Neutron beams have often a considerable portion of gamma rays with fast neutrons. Gamma rays, as beam contaminants, can cause considerable damage to normal tissues even if such tissues do contain high boron concentrations. Materials and Methods: The modification of radio response in human lymphocytes pretreated with boron or gadolinium compound was studied by assessing the DNA damage using single cell gel electrophoresis, the comet assay. The lymphocytes from the human peripheral blood were irradiated with 0, 1, 2 and 4 Gy of gamma rays from a 60 Co isotopic source with or without pretreatment of boron or gadolinium compound for 10 minutes at 4 d egree C . Post-irradiation procedures included slide preparation, cell-lysing, unwinding and electrophoresis, neutralization, staining, and analytic steps, gel electrophoresis. Results: The results indicate that pretreatment with boron compound (50 n M or 250 n M of 10 B) is effective in reducing the radiosensitivity of the lymphocyte DNA. Conversely, pretreatment with gadolinium compound (50 n M) led to a dose-dependent increase in the radiosensitivity, most prominently with a dose of 4 Gy (P<0.001). Furthermore, when the lymphocytes were pretreated with a Combined mixture (1:1) of boron (250 n M) and gadolinium (50 n M) compounds, the reduced radiosensitivity was also observed.

  6. DNA damage in birds after the mining waste spill in southwestern Spain: a Comet assay evaluation.

    Science.gov (United States)

    Pastor, N; López-Lázaro, M; Tella, J L; Baos, R; Forrero, M G; Hiraldo, F; Cortés, F

    2001-01-01

    In April 1998, an ecological disaster resulting from a massive toxic spill of mining acid waste rich in heavy metals posed a serious threat to the Doñana National Park in southwestern Spain. This especially important protected area is the nesting and breeding site for many endangered bird species; white storks (Ciconia ciconia) and black kites (Milvus migrans) are considered the more representative. The suitability of the Comet assay as a biomarker for genotoxic analysis in environmental biomonitoring has been recently validated in studies using different sentinel organisms such as fish, amphibians, rodents, or mollusks. Birds preying on a variety of invertebrate and vertebrate species in the marshlands are appropriate for evaluating the potential deleterious effects of the toxic spill on wildlife of the Dofiana area. Our study on wetland birds high on the aquatic trophic chain sampled within a few months after the toxic spill in the area around Doñana National Park has shown the accumulation of heavy metals. Fourteen months after the mine waste spill, blood samples from white storks and kites collected in the neighborhood of the park and from control birds at reference areas for comparison were examined by fluorescence image analysis after lymphocyte isolation, and by subsequent alkaline single-cell gel (SCG) electrophoresis, known as the Comet assay. Our results indicate that the exposed birds had a significantly increased level of genotoxic damage compared with control animals from noncontaminated locations.

  7. Alkaline gel electrophoresis assay to detect DNA strand breaks and repair mechanisms in Escherichia coli

    Energy Technology Data Exchange (ETDEWEB)

    Mattos, Jose Carlos Pelielo de; Motta, Ellen Serri da; Oliveira, Marcia Betania Nunes de; Dantas, Flavio Jose da Silva; Araujo, Adriano Caldeira de [Universidade do Estado do Rio de Janeiro (UERJ), RJ (Brazil). Dept. de Biofisica e Biometria. Lab. de Radio e Fotobiologia]. E-mail: jcmattos@uerj.br

    2008-12-15

    Reactive oxygen species (ROS) can induce lesions in different cellular targets, including DNA. Stannous chloride (SnCl{sub 2}) is a ROS generator, leading to lethality in Escherichia coli (E. coli), with the base excision repair (BER) mechanism playing a role in this process. Many techniques have been developed to detect genotoxicity, as comet assay, in eukaryotic cells, and plasmid DNA agarose gel electrophoresis. In this study, an adaptation of the alkaline gel electrophoresis method was carried out to ascertain the induction of strand breaks by SnCl{sub 2} in bacterial DNA, from E. coli BER mutants, and its repair pathway. Results obtained show that SnCl{sub 2} was able to induce DNA strand breaks in all strains tested. Moreover, endonuclease IV and exonuclease III play a role in DNA repair. On the whole, data has shown that the alkaline gel electrophoresis assay could be used both for studying DNA strand breaks induction and for associated repair mechanisms. (author)

  8. DNA-repair measurements by use of the modified comet assay

    DEFF Research Database (Denmark)

    Godschalk, Roger W L; Ersson, Clara; Riso, Patrizia

    2013-01-01

    The measurement of DNA-repair activity by extracts from cells or tissues by means of the single-cell gel electrophoresis (comet) assay has a high potential to become widely used in biomonitoring studies. We assessed the inter-laboratory variation in reported values of DNA-repair activity...... on substrate cells that had been incubated with Ro19-8022 plus light to generate oxidatively damaged DNA. Eight laboratories assessed the DNA-repair activity of three cell lines (i.e. one epithelial and two fibroblast cell lines), starting with cell pellets or with cell extracts provided by the coordinating...... laboratory. There was a large inter-laboratory variation, as evidenced by the range in the mean level of repair incisions between the laboratory with the lowest (0.002incisions/10(6)bp) and highest (0.988incisions/10(6)bp) incision activity. Nevertheless, six out of eight laboratories reported the same cell...

  9. Inter-laboratory variation in DNA damage using a standard comet assay protocol

    DEFF Research Database (Denmark)

    Forchhammer, Lykke; Ersson, Clara; Loft, Steffen

    2012-01-01

    There are substantial inter-laboratory variations in the levels of DNA damage measured by the comet assay. The aim of this study was to investigate whether adherence to a standard comet assay protocol would reduce inter-laboratory variation in reported values of DNA damage. Fourteen laboratories ...

  10. 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

  11. Estimates of DNA strand breakage in bottlenose dolphin (Tursiops truncatus leukocytes measured with the Comet and DNA diffusion assays

    Directory of Open Access Journals (Sweden)

    Adriana Díaz

    2009-01-01

    Full Text Available The analysis of DNA damage by mean of Comet or single cell gel electrophoresis (SCGE assay has been commonly used to assess genotoxic impact in aquatic animals being able to detect exposure to low concentrations of contaminants in a wide range of species. The aims of this work were 1 to evaluate the usefulness of the Comet to detect DNA strand breakage in dolphin leukocytes, 2 to use the DNA diffusion assay to determine the amount of DNA strand breakage associated with apoptosis or necrosis, and 3 to determine the proportion of DNA strand breakage that was unrelated to apoptosis and necrosis. Significant intra-individual variation was observed in all of the estimates of DNA damage. DNA strand breakage was overestimated because a considerable amount (~29% of the DNA damage was derived from apoptosis and necrosis. The remaining DNA damage in dolphin leukocytes was caused by factors unrelated to apoptosis and necrosis. These results indicate that the DNA diffusion assay is a complementary tool that can be used together with the Comet assay to assess DNA damage in bottlenose dolphins.

  12. Hemocytes of zebra mussels (Dreissena polymorpha are relevant cells for the monitoring of environmental genotoxicity by the comet assay.

    Directory of Open Access Journals (Sweden)

    Marc Bonnard

    2015-06-01

    Full Text Available The measure of DNA integrity by the single cell gel electrophoresis (SCGE or comet assay is especially recommended for its sensitivity and its capacity for detecting different types of damages. Therefore, it has been applied in environmental genotoxicity in a variety of organisms. It appears today necessary to define both reference and threshold levels of DNA damage, for their application in in situ biomonitoring. However, little is known about the influence of both biological (sex, reproduction status or external (temperature… confounding factors on the measure of DNA damage by the comet assay. These variables need to be taken into account if the robustness of the assay is to be established (Jha, 2008. In the zebra mussel Dreissena polymorpha (recommended as a sentinel species in the evaluation of freshwater quality the measure of DNA damage by the comet assay is mainly performed on hemocytes, which are circulating cells involved in key physiological functions such as immunity, homeostasis, detoxication…. This communication will present and discuss results from an innovative study about the variability of the baseline level of DNA damage in hemocytes of mussels encaged for one year in the canal de l’Aisne à la Marne (Reims, according to their sex and their reproductive status. The sensitivity and the suitability of hemocytes in the evaluation of environmental genotoxicity will also be discussed, referring to observations during a 6 month-exposure of mussels in mesocosms to environmentally realistic concentrations of carbamazepine.

  13. A simple and novel modification of comet assay for determination of bacteriophage mediated bacterial cell lysis.

    Science.gov (United States)

    Khairnar, Krishna; Sanmukh, Swapnil; Chandekar, Rajshree; Paunikar, Waman

    2014-07-01

    The comet assay is the widely used method for in vitro toxicity testing which is also an alternative to the use of animal models for in vivo testing. Since, its inception in 1984 by Ostling and Johansson, it is being modified frequently for a wide range of application. In spite of its wide applicability, unfortunately there is no report of its application in bacteriophages research. In this study, a novel application of comet assay for the detection of bacteriophage mediated bacterial cell lysis was described. The conventional methods in bacteriophage research for studying bacterial lysis by bacteriophages are plaque assay method. It is time consuming, laborious and costly. The lytic activity of bacteriophage devours the bacterial cell which results in the release of bacterial genomic material that gets detected by ethidium bromide staining method by the comet assay protocol. The objective of this study was to compare efficacy of comet assay with different assay used to study phage mediated bacterial lysis. The assay was performed on culture isolates (N=80 studies), modified comet assay appear to have relatively higher sensitivity and specificity than other assay. The results of the study showed that the application of comet assay can be an economical, time saving and less laborious alternative to conventional plaque assay for the detection of bacteriophage mediated bacterial cell lysis. Copyright © 2014 Elsevier B.V. All rights reserved.

  14. Genotoxicity determinations of coriander drop and extract of Coriander Sativum cultured fibroblast of rat embryo by comet assay

    International Nuclear Information System (INIS)

    Heibatullah, K.; Marzieh, P.; Arefeh, I.; Ebrahim, M.

    2008-01-01

    The single cell gel electrophoresis (SCGE) or comet assay is a quick, simple and sensitive technique for measuring DNA damage in cell nucleus. It is well known that medicinal herbs play an important role in the life of human beings, thus it is essential to determine their safety as public health is concerned. In this study the genotoxicity of Coriander drop, herbal pharmaceutical product, and the extract of Coriander sativum were examined in cultured fibroblast of rat embryo using comet assay. The thirteen to fifteen days old rat embryos were lysed with tripsin and after certain steps it was centrifuged and then cultured. After three to five passages, different concentrations of each product were applied to the fibroblasts. Lysing, electrophoresis, neutralization and staining were carried out. Finally the slides were analyzed with fluorescence microscope. In the test groups the results indicated that coriander drop at different doses showed some fragmentation of DNA but this damage as a result was deemed to be not significant. However, in the case of Coriander sativum extract the results showed no mutagenic effects in comparison with the positive control group (p<0.05). In conclusion, these herbal products did not show any magnetic effect according to our test, but further genotoxicity assays are recommended. (author)

  15. DNA Comet Assay and Changes in Microflora Load as Screening Methods to Detect Irradiated Food in Egypt

    International Nuclear Information System (INIS)

    Hammad, A.A.; Abo El Nour, S.A.; Ibrahim, H.M.; Osman, M.E.; Abo El- Nasr, A.

    2014-01-01

    In the present study the microgel electrophoresis of single cells (DNA Comet Assay), and changes in microflora load were applied to detect irradiation treatment of strawberries and fresh-deboned chicken produced in Egypt. Strawberry samples were irradiated at 1.0, 2.0, 3.0 and 4.0 kGy, stored at 4 degree C±1 and analyzed at 0 and 7 days post.-irradiation. Fresh- deboned chicken meat samples were exposed to 2.0, 3.0, 4.0 and 5.0 kGy, stored at 4 degree C±1 and analyzed at 0, 7, 14 and 21 days post-irradiation. After electrophoresis performance, the accridine orange stain slides were seen under fluorescent microscope and the DNA comets were evaluated by photographic and image analysis. Changes in microflora load of irradiated samples were also evaluated. In all irradiated samples, the DNA fragments stretched or migrated out of the cells towards the anode of the agrose gel and appeared as a “comets” with tail. Whereas, DNA comets of all non-irradiated samples were almost intact, round without tail or had very short tail. Values of DNA % in tails and the tail length increased with increasing irradiation dose and storage times. The DNA comet assay could successfully be used to detect radiation treatment of strawberry and deboned-chicken meat samples up to 7 and 21 days post-irradiation, respectively. The absence of gram-negative bacteria and enterobacteriaceae group as well as the very low count of fungi (mostly yeasts) might be considered another evidence of radiation treatment of strawberries and fresh-deboned chicken.

  16. Assessment of gamma ray-induced DNA damage in Lasioderma serricorne using the comet assay

    International Nuclear Information System (INIS)

    Kameya, Hiromi; Miyanoshita, Akihiro; Imamura, Taro; Todoriki, Setsuko

    2012-01-01

    We attempted a DNA comet assay under alkaline conditions to verify the irradiation treatment of pests. Lasioderma serricorne (Fabricius) were chosen as test insects and irradiated with gamma rays from a 60 Co source at 1 kGy. We conducted the comet assay immediately after irradiation and over time for 7 day. Severe DNA fragmentation in L. serricorne cells was observed just after irradiation and the damage was repaired during the post-irradiation period in a time-dependent manner. The parameters of the comet image analysis were calculated, and the degree of DNA damage and repair were evaluated. Values for the Ratio (a percentage determined by fluorescence in the damaged area to overall luminance, including intact DNA and the damaged area of a comet image) of individual cells showed that no cells in the irradiated group were included in the Ratio<0.1 category, the lowest grade. This finding was observed consistently throughout the 7-day post-irradiation period. We suggest that the Ratio values of individual cells can be used as an index of irradiation history and conclude that the DNA comet assay under alkaline conditions, combined with comet image analysis, can be used to identify irradiation history. - Highlights: ► We investigated the DNA comet assay to verify the irradiation of pests. ► Ratio and Tail Moment were higher in irradiated groups than in the control group. ► The DNA comet assay can be used to identify irradiation history.

  17. Comet assay in the detection of irradiated garlic

    International Nuclear Information System (INIS)

    Villavicencio, Anna Lucia C.H.; Marin-Huachaca, Nelida Simona; Romanelli, Maria Fernanda; Delincee, Henry

    2002-01-01

    The increased claim for fresh produce has forced a consensus between nations to pay more attention to the phytosanitary regulations. Inhibition of sprouting of bulbs and tubers by applying ionising radiation is authorised by the National Food Codes in Brazil. The availability of methods for detection of irradiated food will contribute to increase consumers' confidence. A quick and simple screening test to indicate whether a food product has been irradiated or not was utilised in this study. The DNA comet assay was applied to verify whether garlic imported from China had been irradiated or not. This test has already been adopted as a European Standard (EN 13784), for detection of irradiated food. Non-irradiated control samples of garlic and garlic treated with maleic hydrazide were compared with garlic samples irradiated in our department. The unirradiated samples exhibited only limited DNA migration. If samples were irradiated, an increased DNA fragmentation was observed which permitted the discrimination between non-irradiated and irradiated samples. Since the garlic samples from China showed only very limited DNA fragmentation, they were deemed non-irradiated. Thus, this simple screening test was shown to be successful for identification of an irradiation treatment. (author)

  18. A buccal cell model comet assay: Development and evaluation for human biomonitoring and nutritional studies

    Energy Technology Data Exchange (ETDEWEB)

    Szeto, Y.T. [Department of Health Technology and Informatics, Hong Kong Polytechnic University, Kowloon, Hong Kong (China); School of Health Sciences, Macao Polytechnic Institute, Macao (China); Benzie, I.F.F. [Department of Health Technology and Informatics, Hong Kong Polytechnic University, Kowloon, Hong Kong (China)]. E-mail: iris.benzie@inet.polyu.edu.hk; Collins, A.R. [Department of Nutrition, University of Oslo, Oslo (Norway); Choi, S.W. [Department of Health Technology and Informatics, Hong Kong Polytechnic University, Kowloon, Hong Kong (China); Cheng, C.Y. [Department of Health Technology and Informatics, Hong Kong Polytechnic University, Kowloon, Hong Kong (China); Yow, C.M.N. [Department of Health Technology and Informatics, Hong Kong Polytechnic University, Kowloon, Hong Kong (China); Tse, M.M.Y. [School of Nursing, Hong Kong Polytechnic University, Kowloon, Hong Kong (China)

    2005-10-15

    The comet assay is a widely used biomonitoring tool for DNA damage. The most commonly used cells in human studies are lymphocytes. There is an urgent need to find an alternative target human cell that can be collected from normal subjects with minimal invasion. There are some reports of buccal cells, collected easily from the inside of the mouth, being used in studies of DNA damage and repair, and these were of interest. However, our preliminary studies following the published protocol showed that buccal cells sustained massive damage and disintegrated at the high pH [O. Ostling, K.J. Johanson. Microelectrophoretic study of radiation-induced DNA damages in individual mammalian cells. Biochem. Biophys. Res. Commun. 123 (1984) 291-298] used, but that at lower pH were extremely resistant to lysis, an essential step in the comet assay. Therefore, the aims of this study were to develop a protocol than enabled buccal cell lysis and DNA damage testing in the comet assay, and to use the model to evaluate the potential use of the buccal cell model in human biomonitoring and nutritional study. Specifically, we aimed to investigate intra- and inter-individual differences in buccal cell DNA damage (as strand breaks), the effect of in vitro exposure to both a standard oxidant challenge and antioxidant treatment, as well as in situ exposure to an antioxidant-rich beverage and supplementation-related effects using a carotenoid-rich food. Successful lysis was achieved using 0.25% trypsin for 30 min followed by proteinase K (1 mg/ml) treatment for 60 min. When this procedure was performed on cells pre-embedded in agarose on a microscope slide, followed by electrophoresis (in 0.01 M NaOH, 1 mM EDTA, pH 9.1, 18 min at 12 V), a satisfactory comet image was obtained, though inter-individual variation was quite wide. Pre-lysis exposure of cells to a standard oxidant challenge (induced by H{sub 2}O{sub 2}) increased DNA strand breaks in a dose related manner, and incubation of cells in

  19. A buccal cell model comet assay: Development and evaluation for human biomonitoring and nutritional studies

    International Nuclear Information System (INIS)

    Szeto, Y.T.; Benzie, I.F.F.; Collins, A.R.; Choi, S.W.; Cheng, C.Y.; Yow, C.M.N.; Tse, M.M.Y.

    2005-01-01

    The comet assay is a widely used biomonitoring tool for DNA damage. The most commonly used cells in human studies are lymphocytes. There is an urgent need to find an alternative target human cell that can be collected from normal subjects with minimal invasion. There are some reports of buccal cells, collected easily from the inside of the mouth, being used in studies of DNA damage and repair, and these were of interest. However, our preliminary studies following the published protocol showed that buccal cells sustained massive damage and disintegrated at the high pH [O. Ostling, K.J. Johanson. Microelectrophoretic study of radiation-induced DNA damages in individual mammalian cells. Biochem. Biophys. Res. Commun. 123 (1984) 291-298] used, but that at lower pH were extremely resistant to lysis, an essential step in the comet assay. Therefore, the aims of this study were to develop a protocol than enabled buccal cell lysis and DNA damage testing in the comet assay, and to use the model to evaluate the potential use of the buccal cell model in human biomonitoring and nutritional study. Specifically, we aimed to investigate intra- and inter-individual differences in buccal cell DNA damage (as strand breaks), the effect of in vitro exposure to both a standard oxidant challenge and antioxidant treatment, as well as in situ exposure to an antioxidant-rich beverage and supplementation-related effects using a carotenoid-rich food. Successful lysis was achieved using 0.25% trypsin for 30 min followed by proteinase K (1 mg/ml) treatment for 60 min. When this procedure was performed on cells pre-embedded in agarose on a microscope slide, followed by electrophoresis (in 0.01 M NaOH, 1 mM EDTA, pH 9.1, 18 min at 12 V), a satisfactory comet image was obtained, though inter-individual variation was quite wide. Pre-lysis exposure of cells to a standard oxidant challenge (induced by H 2 O 2 ) increased DNA strand breaks in a dose related manner, and incubation of cells in Trolox

  20. Comets

    International Nuclear Information System (INIS)

    Hughes, D.W.

    1982-01-01

    Comets are objects of considerable fascination and this paper reviews the present knowledge of the physical structure of the cometary nucleus, coma and tail, the orbits of comets in the Solar System, the proposed mechanisms of cometary origin, the decay processes suffered by comets, and the ways in which they can be observed from Earth and by spacecraft. (author)

  1. 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.

  2. Fluorescent in situ hybridization on comets: FISH comet.

    Science.gov (United States)

    Shaposhnikov, Sergey; El Yamani, Naouale; Collins, Andrew R

    2015-01-01

    The DNA in eukaryotic cells is organized into loop domains that represent basic structural and functional units of chromatin packaging. The comet assay, a sensitive method for monitoring DNA damage and repair, involves electrophoresis of nucleoids comprising supercoiled DNA attached to the nuclear matrix. Breaks in the DNA relax the supercoiling and allow DNA loops to expand, and on electrophoresis to move towards the anode, giving the appearance of a comet tail. We use fluorescent in situ hybridization (FISH) to investigate the structure of the chromatin within comet preparations and to study specific DNA sequences within comets. In this chapter we describe our FISH comets protocols, deal with some technical questions and outline the theory. FISH with comets should be useful to researchers interested in the structural organization of DNA and chromatin, the localization of DNA damage, and the kinetics of repair of damage.

  3. Using a medium-throughput comet assay to evaluate the global DNA methylation status of single cells

    Directory of Open Access Journals (Sweden)

    Angelique eLewies

    2014-07-01

    Full Text Available The comet assay is a simple and cost effective technique, commonly used to analyse and quantify DNA damage in individual cells. The versatility of the comet assay allows introduction of various modifications to the basic technique. The difference in the methylation sensitivity of the isoschizomeric restriction enzymes HpaII and MspI are used to demonstrate the ability of the comet assay to measure the global DNA methylation level of individual cells when using cell cultures. In the experiments described here, a medium-throughput comet assay and methylation sensitive comet assay are combined to produce a methylation sensitive medium-throughput comet assay to measure changes in the global DNA methylation pattern in individual cells under various growth conditions.

  4. Cytotoxicity and genotoxicity assessment of Euphorbia hirta in MCF-7 cell line model using comet assay.

    Science.gov (United States)

    Ping, Kwan Yuet; Darah, Ibrahim; Chen, Yeng; Sasidharan, Sreenivasan

    2013-09-01

    To evaluate the cytotoxicity and genotoxicity activity of Euphorbia hirta (E. hirta) in MCF-7 cell line model using comet assay. The cytotoxicity of E. hirta extract was investigated by employing brine shrimp lethality assay and the genotoxicity of E. hirta was assessed by using Comet assay. Both toxicity tests exhibited significant toxicity result. In the comet assay, the E. hirta extract exhibited genotoxicity effects against MCF-7 DNA in a time-dependent manner by increasing mean percentage of DNA damage. The extract of E. hirta showed significant toxicity against brine shrimp with an LC₅₀ value of 620.382 µg/mL (24 h). Comparison with positive control potassium dichromate signifies that cytotoxicity exhibited by the methanol extract might have moderate activity. The present work confirmed the cytotoxicity and genotoxicity of E. hirta. However, the observed toxicity of E. hirta extracts needs to be confirmed in additional studies.

  5. Testing the genotoxicity of some perfumes with high diethylphthalate (DEP levels using comet assay

    Directory of Open Access Journals (Sweden)

    Iman Al-Saleh

    2015-05-01

    Full Text Available The presence of phthalates in perfumes has gained recently some attention since these chemicals are added sometimes intentionally as a fixative. Our previous study tested 47 branded perfumes sold in Saudi market and found 68% of tested samples had diethylphthalate (DEP, above reported threshold limit of 1 ppm. Of these phthalates, DEP was found to have the highest mean value (1621.63 ppm. These results enticed us to test the potential genotoxicity of 7 brands in which their DEP contents were in the range of 1.06 ppm to 23649.3 ppm in human TK-6 cells using single cell gel electrophoresis assay (comet assay. Cells were exposed to 400 µl perfume for 2 hrs, at room temperature. Tail moment was (TM used as a metric measure for DNA damage and 25 cells per sample were determined by image analysis software. Four perfumes with DEP above 40 ppm produced significant DNA damage in TK-6 cells with TM of 54.27 ± 1.04, n=500 compare to the other 3 perfumes with DEP < 2 ppm (21.94 ± 1.09, n=300, untreated cells (2.99 ± 0.17, n=250 and cells induced with ethanol (33.76 ± 1.4, n=75 or methanol (23.82 ± 1.84, n=100 which are the vehicles used in perfume's preparations. The results suggest that DEP in perfumes might be one of the ingredients that provoked DNA damage; however, further investigation is required confirming our observation.

  6. Assessment of electron beam-induced DNA damage in larvae of chestnut weevil, Curculio sikkimensis (Heller) (Coleoptera: Curculionidae) using comet assay

    Energy Technology Data Exchange (ETDEWEB)

    Todoriki, Setsuko [Radiation and Information Technology Laboratory, National Food Research Institute, Tsukuba, Ibaraki 305-8642 (Japan)]. E-mail: setsuko@nfri.affrc.go.jp; Hasan, Mahbub [Laboratory for Stored Product Protection, Department of Zoology, Rajshahi University, Rajshahi 6205 (Bangladesh); Miyanoshita, Akihiro [Radiation and Information Technology Laboratory, National Food Research Institute, Tsukuba, Ibaraki 305-8642 (Japan); Imamura, Taro [Radiation and Information Technology Laboratory, National Food Research Institute, Tsukuba, Ibaraki 305-8642 (Japan); Hayashi, Toru [Radiation and Information Technology Laboratory, National Food Research Institute, Tsukuba, Ibaraki 305-8642 (Japan)

    2006-02-15

    Effect of electron beam treatment on DNA damage in mature larvae of chestnut weevil Curculio sikkimensis (Heller) was assessed using single-cell gel electrophoresis (DNA comet assay). Electrons at acceleration voltages of 0 (control), 300, 750, 1000, and 1500 kV at radiation doses of 1 and 4 kGy were used. Electron beam-treated chestnut larvae showed typical DNA fragmentation, compared with cells from non-treated ones which showed a more intact DNA. Investigations using the comet assay showed that the parameters including tail length, tail moment, olive tail moment as well as the quota of DNA damage at both the doses were significantly larger than the control batch larvae. Thus, this technique could contribute to analytical identification of an effective disinfestation and quarantine treatment.

  7. DNA damage and repair assessed by comet assay in workers exposed to lead in a battery recycling.

    Directory of Open Access Journals (Sweden)

    Mahara Valverde

    2015-05-01

    In addition to these findings, DNA damage determined by comet assay was sensible to reflect lead exposure levels related to specific activities inside this factory. Human biomonitoring studies through comet assay could be robust when additional biomarkers are determined at time.

  8. Results of the International Validation of the in vivo rodent alkaline comet assay for the detection of genotoxic carcinogens: Individual data for 1,2-dibromoethane, p-anisidine, and o-anthranilic acid in the 2nd step of the 4th phase Validation Study under the JaCVAM initiative.

    Science.gov (United States)

    Takasawa, Hironao; Takashima, Rie; Narumi, Kazunori; Kawasako, Kazufumi; Hattori, Akiko; Kawabata, Masayoshi; Hamada, Shuichi

    2015-07-01

    As part of the Japanese Center for the Validation of Alternative Methods (JaCVAM)-initiative International Validation Study of an in vivo rat alkaline comet assay, we examined 1,2-dibromoethane (DBE), p-anisidine (ASD), and o-anthranilic acid (ANT) to investigate the effectiveness of the comet assay in detecting genotoxic carcinogens. Each of the three test chemicals was administered to 5 male Sprague-Dawley rats per group by oral gavage at 48, 24, and 3h before specimen preparation. Single cells were collected from the liver and glandular stomach at 3h after the final dosing, and the specimens prepared from these two organs were subjected to electrophoresis under alkaline conditions (pH>13). The percentage of DNA intensity in the comet tail was then assessed using an image analysis system. A micronucleus (MN) assay was also conducted using these three test chemicals with the bone marrow (BM) cells collected from the same animals simultaneously used in the comet assay, i.e., combination study of the comet assay and BM MN assay. A genotoxic (Ames positive) rodent carcinogen, DBE gave a positive result in the comet assay in the present study, while a genotoxic (Ames positive) non-carcinogen, ASD and a non-genotoxic (Ames negative) non-carcinogen, ANT showed negative results in the comet assay. All three chemicals produced negative results in the BM MN assay. While the comet assay findings in the present study were consistent with those obtained from the rodent carcinogenicity studies for the three test chemicals, we consider the positive result in the comet assay for DBE to be particularly meaningful, given that this chemical produced a negative result in the BM MN assay. Therefore, the combination study of the comet assay and BM MN assay is a useful method to detect genotoxic carcinogens that are undetectable with the BM MN assay alone. Copyright © 2015 Elsevier B.V. All rights reserved.

  9. An ECVAG inter-laboratory validation study of the comet assay

    DEFF Research Database (Denmark)

    Ersson, Clara; Møller, Peter; Forchhammer, Lykke

    2013-01-01

    The alkaline comet assay is an established, sensitive method extensively used in biomonitoring studies. This method can be modified to measure a range of different types of DNA damage. However, considerable differences in the protocols used by different research groups affect the inter-laboratory...

  10. Comet assay as a human biomonitoring tool: application in occupational exposure to antineoplastic drugs

    Directory of Open Access Journals (Sweden)

    Carina Ladeira

    2015-05-01

    Occupational exposure to antineoplastic drugs is associated with genotoxic effects, although comet assay analyzed parameters were higher in exposed comparing with controls, were not significant. Also the study of the susceptibility biomarkers did not show statistical significant differences, the small size of our sample hampered the finding of a possible association, let alone a causality relationship.

  11. Genotoxicity of cadmium in marine diatom Chaetoceros tenuissimus using the alkaline Comet assay

    Digital Repository Service at National Institute of Oceanography (India)

    Desai, S.R.; Verlecar, X.N.; Nagarajappa; Goswami, U.

    Genotoxic effects of Cadmium on phytoplankton Chaetoceros tenuissimus have been evaluated using DNA damage by comet assay. Cadmium concentrations ranging from 2.4 to 10 mg/L were used to evaluate the effects. Results showed that as the concentration...

  12. In vivo Comet assay – statistical analysis and power calculations of mice testicular cells

    DEFF Research Database (Denmark)

    Hansen, Merete Kjær; Sharma, Anoop Kumar; Dybdahl, Marianne

    2014-01-01

    -97-5, CAS no. 85-28-9, CAS no. 13674-87-8, CAS no. 43100-38-5 and CAS no. 60965-26-6. Testicular cells were examined using the alkaline version of the Comet assay and the DNA damage was quantified as % tail DNA using a fully automatic scoring system. From the raw data 23 summary statistics were examined...

  13. How to best freeze liver samples to perform the in vivo mammalian alkaline comet assay

    Directory of Open Access Journals (Sweden)

    José Manuel Enciso Gadea

    2015-06-01

    None of the different methods used was capable of giving good results, except immersing the liver samples in liquid nitrogen, followed by Jackson’s et al. (2013 thawing protocol, suggesting that the thawing process may be as critical as the freezing process. To sum up, these results highlight the importance of deepening the possibility to perform the comet assay with frozen tissue.

  14. Analysis of lipoproteins by capillary zone electrophoresis in microfluidic devices: Assay development and surface roughness measurements

    NARCIS (Netherlands)

    Weiller, Bruce H.; Ceriotti, Laura; Shibata, Takayuki; Rein, Dietrich; Roberts, Matthew A.; Lichtenberg, Jan; German, J. Bruce; De Rooij, Nico F.; Verpoorte, Elisabeth

    2002-01-01

    The development of a new assay for lipoproteins by capillary electrophoresis in fused-silica capillaries and in glass microdevices is described in this paper. The separation of low-density (LDL) and high-density (HDL) lipoproteins by capillary zone electrophoresis is demonstrated in fused-silica

  15. Identification of radiation treatment of foods using novel technique of 'DNA comet assay'

    International Nuclear Information System (INIS)

    Khan, A.A.; Khan, H.M.; Wasim, M.A.

    2005-01-01

    Treatment of food using ionizing radiation is being progressively used in many countries to inactivate food pathogens, to eradicate pests and to extend shelf life thereby contributing to safer and more plentiful food supply. Food control agencies throughout the world need some reliable, simple and rapid methods for the detection foods to ensure free choice of consumer and to enforce labeling. The DNA comet assay offers great potential as a rapid tool to screen irradiated and unirradiated samples of several kinds of foods. In the present study samples of fresh and frozen beef has investigated for the detection of irradiation treatment. The samples were subjected to radiation doses of 0,4.5 and 7 KGy and were stored in freezer before analysis. The cells were extracted into cold PBS solutions, embedded into agarose gel on microscope slides, lysed and eletrophoressed at a voltage of 2V/cm for 2 minutes. The fragmented DNA as a irradiation treatment was stretched in the gel producing the dose dependent comets. These comets were visible using a simple transmission microscope after silver staining. The controlled and irradiation samples of meat were clearly distinguishable on the basis of the stained patterns of DNA in from of round or conical intact cells for unirradiated samples or in from of comets for irradiated samples. It is therefore concluded that DNA comet Assay offers a potential to screen unirradiated and irradiated meat samples. (author)

  16. Detection of radiation treatment of meat by novel techniques of DNA comet assay

    International Nuclear Information System (INIS)

    Khan, A.A.; Khan, H.M.

    2002-01-01

    Treatment of food to ionizing radiation is being progressively used in many countries to inactivate food pathogens, to eradicate pests and to extend shelf life; thereby contributing to safer and more plentiful food supply. Food control agencies throughout the world need some reliable, simple and rapid methods for detection of irradiated foods to ensure free choice of consumer and to enforce labeling. The DNA comet assay offers great potential as a rapid tool to screen irradiated and unirradiated samples of several kinds of foods. In the present study, frozen beef has been investigated for detection of irradiation treatment. The samples were subjected to radiation doses of 0,4,5 and 7.0 kGy and were stored in freezer before analysis. The cells were extracted into cold PBS solutions, embedded into the agarose gel on microscope slides, lysed and electrophoressed at a voltage of 2v/cm for 2 min. The fragmented DNA as a result of irradiation treatment was stretched in the gel producing the dose dependent comets. These comets were visible using a simple transmission microscope after silver staining. The controlled and irradiated samples of meat were clearly distinguishable on the basis of the stained patterns of DNA in form of round or conical intact cells for unirradiated samples or in form of comets for irradiated samples. It is therefore, concluded that 'DNA Comet Assay' offers a potential to screen unirradiated and irradiated meat samples. (author)

  17. Chromatin in fractal globule state: evidence from comet assay

    Directory of Open Access Journals (Sweden)

    Afanasieva K. S.

    2015-04-01

    Full Text Available At higher order levels chromatin is organized into loops that appear as a result of contacts between distant loci. The aim of this work was to investigate the length distribution of the DNA loops in nucleoids obtained after lysis of either whole cells or isolated cell nuclei. Methods. We used single cell gel electrophoresis to analyze the kinetics of the DNA loop migration from the two nucleoid types. Results. The kinetics of the DNA exit was found to have specific features for the two types of nucleoids. At the same time, in both cases, the DNA amount in the electrophoretic track depends linearly on the length of the longest loops in the track. Conclusions. We have concluded that for the loops up to ~ 100 kb the length distribution appears to be consistent with the fractal globule organization.

  18. Genotoxicity of cadmium chloride in the marine gastropod Nerita chamaeleon using comet assay and alkaline unwinding assay.

    Science.gov (United States)

    Sarkar, Anupam; Bhagat, Jacky; Ingole, Baban S; Rao, Durga P; Markad, Vijaykumar L

    2015-02-01

    This paper presents an evaluation of the genotoxic effects of cadmium chloride (CdCl2 ) on marine gastropod, Nerita chamaeleon following the technique of comet assay and the DNA alkaline unwinding assay (DAUA). In this study, the extent of DNA damage in gill cells of N. chamaeleon was measured after in vivo exposure to four different concentrations (10, 25, 50, and 75 µg/L) of CdCl2 . In vitro exposure of hydrogen peroxide (H2 O2 ; 1, 10, 25, and 50 µM) of the gill cells showed a significant increase in the percentage tail DNA, Olive tail moment, and tail length (TL). Significant changes in percentage tail DNA by CdCl2 exposure were observed in all exposed groups of snails with respect to those in control. Exposure to 75 µg/L of CdCl2 produced significant decrease in DNA integrity as measured by DAUA at all duration with respect to control. In vivo exposure to different concentrations of CdCl2 (10, 25, 50, and 75 µg/L) to N. chamaeleon showed considerable increase in DNA damage as observed by both alkaline comet assay and the DAUA. The extent of DNA damage in marine gastropods determined by the application of alkaline comet assay and DAUA clearly indicated the genotoxic responses of marine gastropod, N. chamaeleon to a wide range of cadmium concentration in the marine environment. © 2013 Wiley Periodicals, Inc.

  19. DNA alkylation lesions and their repair in human cells: modification of the comet assay with 3-methyladenine DNA glycosylase (AlkD).

    Science.gov (United States)

    Hašplová, Katarína; Hudecová, Alexandra; Magdolénová, Zuzana; Bjøras, Magnar; Gálová, Eliška; Miadoková, Eva; Dušinská, Mária

    2012-01-05

    3-methyladenine DNA glycosylase (AlkD) belongs to a new family of DNA glycosylases; it initiates repair of cytotoxic and promutagenic alkylated bases (its main substrates being 3-methyladenine and 7-methylguanine). The modification of the comet assay (single cell gel electrophoresis) using AlkD enzyme thus allows assessment of specific DNA alkylation lesions. The resulting baseless sugars are alkali-labile, and under the conditions of the alkaline comet assay they appear as DNA strand breaks. The alkylating agent methyl methanesulfonate (MMS) was used to induce alkylation lesions and to optimize conditions for the modified comet assay method with AlkD on human lymphoblastoid (TK6) cells. We also studied cellular and in vitro DNA repair of alkylated bases in DNA in TK6 cells after treatment with MMS. Results from cellular repair indicate that 50% of DNA alkylation is repaired in the first 60 min. The in vitro repair assay shows that while AlkD recognises most alkylation lesions after 60 min, a cell extract from TK6 cells recognises most of the MMS-induced DNA adducts already in the first 15 min of incubation, with maximum detection of lesions after 60 min' incubation. Additionally, we tested the in vitro repair capacity of human lymphocyte extracts from 5 individuals and found them to be able to incise DNA alkylations in the same range as AlkD. The modification of the comet assay with AlkD can be useful for in vitro and in vivo genotoxicity studies to detect alkylation damage and repair and also for human biomonitoring and molecular epidemiology studies. Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

  20. OXIDATIVE EFFECTS OF DIFFERENT DIETARY FATS ON MOUSE DNA DETECTED BY COMET ASSAY

    Directory of Open Access Journals (Sweden)

    Romana Marinšek Logar

    2000-06-01

    Full Text Available Here we report on the introduction of the Comet assay for the in vivo testing of oxidative effects of different polyunsaturated dietary fats on mouse nuclear DNA. Five groups of mice were fed with the diet of the same composition but with different fat supplementation: 1st group - oleic acid rich sunflower oil, 2 nd group - sunflower oil, 3rd gruop - rape oil, 4th group - lard, 5th group - control group. Fats with different degrees of unsaturation have caused different degrees of mice DNA damage. The highest degree of DNA damage (2.72 was found in the group fed with oleic acid rich sunflower oil. The lowest degree of DNA damage was found in the control group. The results obtained indicate that Comet assay is sensitive enough to differentiate the quality of nutritional fats from the view of free radicals formation.

  1. Silver (nano)materials cause genotoxicity in Enchytraeus crypticus, as determined by the comet assay

    DEFF Research Database (Denmark)

    Maria, Vera L; Ribeiro, Maria João; Guilherme, Sofia

    2018-01-01

    d the effects were either reduced or repaired, whereas Ag NM300K caused higher genotoxicity after 7d for the lowest concentration (EC20), highlighting a potential non-monotone dose-response effect. Overall, the comet assay showed the power to discriminate effects between materials and also toxicity...... not been used; hence, the goals of the current study included the optimization of the in vivo alkaline comet assay in Enchytraeus crypticus. Further, the effect of silver nanomaterial (Ag NM300K, dispersed, 15 nm) was tested and compared with AgNO3 . Hydrogen peroxide (H2 O2 ) was used as a positive...... control. The various steps were optimized. The fully detailed standard operating procedure (SOP) is presented here. Ag materials caused genotoxicity, this being differentiated for the nano and non-nano forms. AgNO3 caused genotoxicity after 3 days (d) of exposure in a dose related manner, although after 7...

  2. Lack of genotoxicity of formocresol, paramonochlorophenol, and calcium hydroxide on mammalian cells by comet assay.

    Science.gov (United States)

    Ribeiro, Daniel Araki; Marques, Mariângela Esther Alencar; Salvadori, Daisy Maria Fávero

    2004-08-01

    Formocresol, paramonochlorophenol, and calcium hydroxide are widely used in dentistry because of their antibacterial activities in root canal disinfection. However, the results of genotoxicity studies using these materials are inconsistent in literature. The goal of this study was to examine the genotoxic potential of formocresol, paramonochlorophenol, and calcium hydroxide using mouse lymphoma cells and human fibroblasts cells in vitro by the comet assay. Data were assessed by Kruskal-Wallis nonparametric test. The results showed that all compounds tested did not cause DNA damage for the tail moment or tail intensity parameters. These findings suggest that formocresol, paramonochlorophenol, and calcium hydroxide do not promote DNA damage in mammalian cells and that the comet assay is a suitable tool to investigate genotoxicity.

  3. The DNA comet assay and the germination test in detection of food treated by ionizing radiation

    International Nuclear Information System (INIS)

    Huachaca, Nelida Simona Marin

    2002-01-01

    Two methods of irradiated food detection, one biochemical, the comet assay and, other biological, the germination test, were applied in bovine meat and fruit samples. The comet assay detects the damage on DNA caused by ionizing radiation. The germination test evaluates the sensitivity to radiation of seeds as for germination ability, shooting and, rooting. The samples were irradiated in gamma font and electron accelerator. For bovine meat samples, the doses were 0.0; 2.5; 4.5 e 7.0 kGy at chilled condition and, 0.0; 2.5; 4.5; 7.0 e 8.5 kGy at frozen conditions. For fruit samples such as melon, watermelon, apple, orange, papaya and, tomato, the doses were: 0.0; 0.5; 0.75; 1.0; 2.0 e 4.0 kGy. The differences between the gamma rays and the electron beam effects on extent of DNA migration and, on shooting and rooting, showed to be similar. The comet assay, under neutral conditions, permitted to discriminate between irradiated and unirradiated bovine meat samples, until one month of storage. Also, it was possible to distinguish, by the comet assay, the control sample with regard to irradiated fruit, at doses as low as 0,5 kGy. In the germination test, the root length was the best parameter to discriminate irradiated and unirradiated samples of melon, watermelon and tomato, while the germination percent was the best parameter for apple and orange. (author)

  4. Evaluation of the neutral comet assay for detection of alpha-particle induced DNA-double-strand-breaks; Evaluation des Comet Assays bei neutralem pH zur Detektion von α-Partikel induzierten DNA-Doppelstrangbruechen

    Energy Technology Data Exchange (ETDEWEB)

    Hofbauer, Daniela

    2010-10-20

    Aim of this study was to differentiate DNA-double-strand-breaks from DNA-single-strand-breaks on a single cell level, using the comet assay after α- and γ-irradiation. Americium-241 was used as a alpha-irradiation-source, Caesium-137 was used for γ-irradiation. Because of technical problems with both the neutral and alkaline comet assay after irradiation of gastric cancer cells and human lymphocytes, no definite differentiation of DNA-damage was possible.

  5. Genotoxicity evaluation of dental restoration nanocomposite using comet assay and chromosome aberration test.

    Science.gov (United States)

    Musa, Marahaini; Ponnuraj, Kannan Thirumulu; Mohamad, Dasmawati; Rahman, Ismail Ab

    2013-01-11

    Nanocomposite is used as a dental filling to restore the affected tooth, especially in dental caries. The dental nanocomposite (KelFil) for tooth restoration used in this study was produced by the School of Dental Sciences, Universiti Sains Malaysia, Malaysia and is incorporated with monodispersed, spherical nanosilica fillers. The aim of the study was to determine the genotoxic effect of KelFil using in vitro genotoxicity tests. The cytotoxicity and genotoxicity of KelFil was evaluated using MTT assay, comet assay and chromosome aberration tests with or without the addition of a metabolic activation system (S9 mix), using the human lung fibroblast cell line (MRC-5). Concurrent negative and positive controls were included. In the comet assay, no comet formation was found in the KelFil groups. There was a significant difference in tail moment between KelFil groups and positive control (p < 0.05). Similarly, no significant aberrations in chromosomes were noticed in KelFil groups. The mitotic indices of treatment groups and negative control were significantly different from positive controls. Hence, it can be concluded that the locally produced dental restoration nanocomposite (KelFil) is non-genotoxic under the present test conditions.

  6. The use of the comet assay in the study of human nutrition and cancer.

    Science.gov (United States)

    Wasson, Gillian R; McKelvey-Martin, Valerie J; Downes, C Stephen

    2008-05-01

    The influence of diet on carcinogenesis is a hugely complex area; not only is the consumption of major dietary factors such as meat, fat and fruits and vegetables associated with increased or decreased risk of a range of cancers but also an increasing number of specific nutrients such as vitamins, minerals and phytochemicals are being proposed as the next 'superfoods' to combat the development of cancer. As well as epidemiological studies to determine the association of these dietary factors with cancer risk, it is also essential to investigate the underlying mechanisms through which these factors may causally influence carcinogenesis. The comet assay provides a relatively simple, cheap and rapid method to examine DNA damage and repair and is, therefore, an ideal biomarker for the study of the effects of nutrition on cancer. This review focuses on the use of the comet assay in studies involving human subjects or human cell lines, which investigate the effects of various nutrients on biomarkers relevant to carcinogenesis, and discusses the potential of the comet assay and its various modifications for use as cancer-related biomarkers suitable for use in nutritional studies.

  7. Effects of photopolymerisation on genotoxicity of composite adhesives in the comet assay

    Directory of Open Access Journals (Sweden)

    Dačić Stefan

    2016-01-01

    Full Text Available Certain in vivo studies have shown that the application of adhesives directly onto the open pulp or on a thin layer of dentin causes inflammation and pulpal abscesses. This reaction is related to toxic effects of monomers from adhesives. It has been confirmed that after proper illumination the adhesives become less toxic. The aim of the study was to examine genotoxicity of non-polymerised, partly polymerised and polymerised adhesives on isolated human lymphocytes using the alkaline Comet assay. Adper Single bond2 and Adper Easy One/3M ESPE adhesive photopolymerisation was performed by Elipar Highlight 3M ESPE halogen lamp for 0, 10 and 40 sec, at final concentrations of 100, 200, 500 and 1000 μg/mL. With both adhesives, photopolymerisation at 0 and 10 seconds showed statistically significant increase in DNA damage in comparision to the negative control (solvent. On the other hand, after 40 seconds of photopolymerisation of both adhesives in all tested concentrations, the degree of DNA damage in Comet assay had no significant difference (P>0.05, χ2 test compared to the negative control. Therefore, only the 40 seconds of photopolymerisation prevented genotoxic effects of both adhesives in the Comet assay. [Projekat Ministarstva nauke Republike Srbije, br. III46002 i br. OI173012

  8. Genotoxicity evaluation of dental restoration nanocomposite using comet assay and chromosome aberration test

    Science.gov (United States)

    Musa, Marahaini; Thirumulu Ponnuraj, Kannan; Mohamad, Dasmawati; Rahman, Ismail Ab

    2013-01-01

    Nanocomposite is used as a dental filling to restore the affected tooth, especially in dental caries. The dental nanocomposite (KelFil) for tooth restoration used in this study was produced by the School of Dental Sciences, Universiti Sains Malaysia, Malaysia and is incorporated with monodispersed, spherical nanosilica fillers. The aim of the study was to determine the genotoxic effect of KelFil using in vitro genotoxicity tests. The cytotoxicity and genotoxicity of KelFil was evaluated using MTT assay, comet assay and chromosome aberration tests with or without the addition of a metabolic activation system (S9 mix), using the human lung fibroblast cell line (MRC-5). Concurrent negative and positive controls were included. In the comet assay, no comet formation was found in the KelFil groups. There was a significant difference in tail moment between KelFil groups and positive control (p treatment groups and negative control were significantly different from positive controls. Hence, it can be concluded that the locally produced dental restoration nanocomposite (KelFil) is non-genotoxic under the present test conditions.

  9. Genotoxicity evaluation of dental restoration nanocomposite using comet assay and chromosome aberration test

    International Nuclear Information System (INIS)

    Musa, Marahaini; Ponnuraj, Kannan Thirumulu; Mohamad, Dasmawati; Rahman, Ismail Ab

    2013-01-01

    Nanocomposite is used as a dental filling to restore the affected tooth, especially in dental caries. The dental nanocomposite (KelFil) for tooth restoration used in this study was produced by the School of Dental Sciences, Universiti Sains Malaysia, Malaysia and is incorporated with monodispersed, spherical nanosilica fillers. The aim of the study was to determine the genotoxic effect of KelFil using in vitro genotoxicity tests. The cytotoxicity and genotoxicity of KelFil was evaluated using MTT assay, comet assay and chromosome aberration tests with or without the addition of a metabolic activation system (S9 mix), using the human lung fibroblast cell line (MRC-5). Concurrent negative and positive controls were included. In the comet assay, no comet formation was found in the KelFil groups. There was a significant difference in tail moment between KelFil groups and positive control (p < 0.05). Similarly, no significant aberrations in chromosomes were noticed in KelFil groups. The mitotic indices of treatment groups and negative control were significantly different from positive controls. Hence, it can be concluded that the locally produced dental restoration nanocomposite (KelFil) is non-genotoxic under the present test conditions. (paper)

  10. Evaluation of the neutral comet assay for detection of alpha-particle induced DNA-double-strand-breaks

    International Nuclear Information System (INIS)

    Hofbauer, Daniela

    2010-01-01

    Aim of this study was to differentiate DNA-double-strand-breaks from DNA-single-strand-breaks on a single cell level, using the comet assay after α- and γ-irradiation. Americium-241 was used as a alpha-irradiation-source, Caesium-137 was used for γ-irradiation. Because of technical problems with both the neutral and alkaline comet assay after irradiation of gastric cancer cells and human lymphocytes, no definite differentiation of DNA-damage was possible.

  11. The use of comet assay to assess DNA integrity of boar spermatozoa following liquid preservation at 5 degrees C and 16 degrees C.

    Directory of Open Access Journals (Sweden)

    J Strzezek

    2004-03-01

    Full Text Available The comet assay, under neutral conditions, allows the assessment of DNA integrity influenced by sperm ageing, which is manifested in DNA double-strand breaks. Here, we attempted to use a modified neutral comet assay test (single-cell gel electrophoresis, to our knowledge for the first time, to assess DNA integrity of boar spermatozoa during liquid storage for 96 h at 5 degrees C and 16 degrees C. In this comet assay protocol we used 2% beta-mercaptoethanol prior to the lysis procedure, to aid in removing nuclear proteins. Ejaculates from 3 boars (designated A, C and G were diluted with a standard semen extender, Kortowo-3 (K-3, which was supplemented with lipoprotein fractions extracted from hen egg yolk (LPFh or ostrich egg yolk (LPFo. Irrespective of the extender type, the percentage of comet-detected spermatozoa with damaged DNA increased gradually during prolonged storage at 5 degrees C and 16 degrees C. Spermatozoa stored in K-3 extender exhibited elevated levels of DNA damage at both storage temperatures. Significant differences in DNA damage among the boars were more pronounced during storage in LPF-based extenders at 5 degrees C: spermatozoa of boars A and G were less susceptible to DNA damage. The percent of tail DNA in comets was lower in LPF-based extenders, and there were individual variations among the boars. We observed that changes in DNA integrity were dependent on the extender type and storage temperature. A higher level of DNA instability was observed in K-3 extended semen compared with K-3/LPFh or K-3/LPFo extended semen during storage at 5 degrees C. No significant difference in the level of DNA damage between K-3/LPFh and K-3/LPFo was observed. It seems that a long-term storage can affect genomic integrity of boar spermatozoa. The modified neutral comet assay can be used to detect low levels of DNA damage in boar spermatozoa during liquid preservation. Therefore, screening for sperm DNA damage may be used as an additional

  12. Evaluation through comet assay of DNA damage induced in human lymphocytes by alpha particles. Comparison with protons and Co-60 gamma rays

    International Nuclear Information System (INIS)

    Di Giorgio, M.; Kreiner, A. J.; Schuff, J. A.; Vallerga, M. B.; Taja, M. R.; Lopez, F. O.; Alvarez, D. E.; Saint Martin, G.; Burton, A.; Debray, M. E.; Kesque, J. M.; Somacal, H.; Stoliar, P.; Valda, A.; Davidson, J.; Davidson, M.; Ozafran, M. J.; Vazquez, M. E.

    2004-01-01

    Several techniques with different sensitivity to single-strand breaks and/or double strand breaks were applied to detect DNA breaks generated by high LET particles. Tests that assess DNA damage in single cells might be the appropriate tool to estimate damage induced by particles, facilitating the assessment of heterogeneity of damage in a cell population. The microgel electrophoresis (comet) assay is a sensitive method for measuring DNA damage in single cells. The objective of this work was to evaluate the proficiency of comet assay to assess the effect of high LET radiation on peripheral blood lymphocytes, compared to protons and Co-60 gamma rays. Materials and methods: Irradiations of blood samples were performed at TANDAR laboratory (Argentina). Thin samples of human peripheral blood were irradiated with different doses (0-2.5 Gy) of 20.2 MeV helium-4 particles in the track segment mode, at nearly constant LET. Data obtained were compared with the effect induced by a MeV protons and Co-60 gamma rays. Alkaline comet assay was applied. Comets were quantified by the Olive tail moment. Distribution of the helium-4 particle and protons were evaluated considering Poisson distribution in lymphocyte nuclei. The mean dose per nucleus per particle result 0.053 Gy for protons and 0.178 Gy for helium-4 particles. When cells are exposed to a dose of 0.1 Gy, the hit probability model predicts that 43% of the nuclei should have experienced and alpha traversal while with protons, 85% of the nuclei should be hit. The experimental results show a biphasic response for helium-4 particles (0.1 Gy), indicating the existence of two subpopulations: unhit and hit. Distributions of tail moment as a function of fluence and experimental dose for comets induced by helium-4 particles, protons and Co-60 gamma rays were analyzed. With helium-4 irradiations, lymphocyte nuclei show an Olive tail moment distribution flattened to higher tail moments a dose increase. However, for irradiations with

  13. Comet assay as a procedure for detecting possible genotoxicity induced by non-ionizing radiation

    Directory of Open Access Journals (Sweden)

    Zsuzsanna Nemeth

    2015-05-01

    In our laboratory we use comet assay for testing genotoxicity of non-ionizing radiation for more than ten years. In the experiments we use whole blood samples (human or dog, cell lines (e.g. H295R cell line or 3 dimensional in vitro skin tissue (epidermis models. In our protocol a slightly modified alkaline Comet assay method of Singh et al. (1988 is used. On our poster there will be presented a brief summary of our experiments with exposure to different types of radiation (ELF, RF, and intermediate frequency. In our protocols the non-ionizing radiation was often combined with ionizing radiation to see whether the non-ionizing radiation can influence the repair of the DNA damage induced by ionizing radiation. For the evaluation of the slides mainly Komet 4.0 image analysis system software (Kinetic Imaging, Liverpool, UK was used, but as we got familiarized with other methods for slide evaluation like grading the comets by visual scoring into 5 categories or the CaspLab software, the comparison of these three methods will be also presented.

  14. Application of the comet assay in erythrocytes of Oreochromis niloticus (Pisces: a methodological comparison

    Directory of Open Access Journals (Sweden)

    Cintya A. Christofoletti

    2009-01-01

    Full Text Available The present study applied the comet assay to erythrocytes of Oreochromis niloticus with the aim of improving protocols to detect DNA damage in these cells, by using two distinct pHs (pH = 12.1 and pH > 13 and evaluating whether there is a correspondence between silver and ethidium bromide staining. Comets were visually examined and, the frequency of cells with and without damage was obtained, as well as the distribution of classes and scores. By using the Kruskal-Wallis test, our results revealed that pH 12.1 is more effective, although both pHs can be used. Our findings also suggest that silver staining can substitute ethidium bromide, an expensive and highly toxic stain that requires specific equipment for examination.

  15. DNA damage in grasshoppers' larvae--comet assay in environmental approach.

    Science.gov (United States)

    Augustyniak, Maria; Orzechowska, Helena; Kędziorski, Andrzej; Sawczyn, Tomasz; Doleżych, Bogdan

    2014-02-01

    The comet assay that provides a quantitative measure of the DNA-strand breaks may be used for assessing the 'genotoxic potential' of the environment. Young adults of Chorthippus brunneus (Orthoptera), collected at three sites in Southern Poland, differing in the level of pollution, particularly with heavy metals: Pilica (reference), Olkusz (moderately polluted) and Szopienice (heavily polluted) - were allowed to mate under laboratory conditions that were free from any pollution. Egg-pods were collected and, after diapause, brain cells from one-day old larvae were used for the comet assay. We compared the level of DNA damage in the larvae originating from these sites and also measured time-dependent DNA repair after single 10min. application of H2O2 (20μM final concentration). The DNA damage was relatively low in larval cells irrespectively of the site pollution their parents came from. However, measured comet parameters - tail DNA content (TDNA), tail length (TL), and olive tail moment (OTM) - were significantly higher in larvae originating from the Szopienice site than in those from the reference site. Incubation of cells with H2O2 resulted in significantly higher values of the comet parameters in the insects from all the study sites with the highest ones observed in the offspring of grasshoppers from Szopienice. Moreover, DNA repair, following the treatment, did not occur in the latter group. These data contribute to almost unexplored subject of genotoxic effects of environmental pollutants in insects. They are discussed in the light of the concept of adaptive strategies in energy allocation depending on the level of biotope pollution. Copyright © 2013 Elsevier Ltd. All rights reserved.

  16. Capillary electrophoresis methods for microRNAs assays: A review

    Energy Technology Data Exchange (ETDEWEB)

    Ban, Eunmi; Song, Eun Joo, E-mail: ejsong@kist.re.kr

    2014-12-10

    Highlights: • A review of CE analysis of miRNAs. • Summary of developments and applications of CE systems in miRNA studies. • Applications and development of microchip-based CE for rapid analysis of miRNA. - Abstract: MicroRNAs (miRNAs) are short noncoding RNAs that conduct important roles in many cellular processes such as development, proliferation, differentiation, and apoptosis. In particular, circulating miRNAs have been proposed as biomarkers for cancer, diabetes, cardiovascular disease, and other illnesses. Therefore, determination of miRNA expression levels in various biofluids is important for the investigation of biological processes in health and disease and for discovering their potential as new biomarkers and drug targets. Capillary electrophoresis (CE) is emerging as a useful analytical tool for analyzing miRNA because of its simple sample preparation steps and efficient resolution of a diverse size range of compounds. In particular, CE with laser-induced fluorescence detection is a promising and relatively rapidly developing tool with the potential to provide high sensitivity and specificity in the analysis of miRNAs. This paper covers a short overview of the recent developments and applications of CE systems in miRNA studies in biological and biomedical areas.

  17. Individual sensitivity to radiations and DNA repair proficiency: the comet assay contribution

    International Nuclear Information System (INIS)

    Alapetite, C.

    1998-01-01

    Some are hereditary syndromes demonstrate high cancer risk and hypersensitivity in response to exposures to agents such as ultraviolet or ionising radiation, and are characterized by a defective processing of DNA damage. They highlight the importance of the individual risk associated to exposures. The comet assay, a simple technique that detects DNA strand breaks, requires few cells and allows examination of DNA repair capacities in established cell lines, in blood samples or biopsies. The assay has been validated on cellular systems with known repair defects such as xeroderma pigmentosum defective in nucleotide excision repair, on mutant rodent cell lines defective in DNA single strand breaks rejoining (XRCC5/Ku80 and XRCC7/DNAPKcs) (neutral conditions). This assay does not allow to distinguish a defective phenotype in ataxia telangiectasia cells. It shows in homozygous mouse embryo fibroblasts Brca2-/- an impaired DNA double strand break rejoining. Simplicity, rapidity and sensitivity of the alkaline comet assay allow to examine the response of lymphocytes. It has been applied to the analysis of the role of DNA repair in the pathogenesis of collagen diseases, and the involvement of individual DNA repair proficiency in the thyroid tumorigenesis induced in some patients after therapeutic irradiation at childhood has been questioned. Preliminary results of these studies suggest that this type of approach could help for adapting treatment modalities and surveillance in subgroups of patients defective in DNA repair process. It could also have some incidence in the radioprotection field. (author)

  18. Detection of hypoxic fractions in murine tumors by comet assay: Comparison with other techniques

    International Nuclear Information System (INIS)

    Hu, Q.; Kavanagh, M.C.; Newcombe, D.

    1995-01-01

    The alkaline comet assay was used to detect the hypoxic fractions of murine tumors. A total of four tumor types were tested using needle aspiration biopsies taken immediately after a radiation dose of 15 Gy. Initial studies confirmed that the normalized tail moment, a parameter reflecting single-strand DNA breaks induced by the radiation, was linearly related to radiation dose. Further, it was shown that for a mixed population (1:1) of cells irradiated under air-breathing or hypoxic conditions, the histogram of normal tail moment values obtained from analyzing 400 cells in the population had a double peak which, when fitted with two Gaussian distributions, gave a good estimate of the proportion of the two subpopulations. For the four tumor types, the means of the calculated hypoxic fractions from four or five individual tumors were 0.15 ± 0.04 for B16F1, 0.08 ± 0.04 for KHT-LP1, 0.17 ± 0.04 for RIF-1 and 0.04 ± 0.01 for SCCVII. Analysis of variance showed that the hypoxic fraction in KHT-LP1 tumors is significantly lower than those of the other three tumors (P = 0.026) but that there is no significant difference in hypoxic fraction between B16F1, RIF-1 and SCCVII tumors (P = 0.574). Results from multiple samples taken from each of five RIF-1 tumors showed that the intertumor heterogeneity of hypoxic fractions was greater than that within the same tumor. The mean hypoxic fraction obtained using the comet assay for the four tumor types was compared with the hypoxic fraction determined by the clonogenic assay, or median pO 2 values, or [ 3 H]misonidazole binding in the same tumor types. The values of hypoxic fraction obtained with the comet assay were two to four times lower than those measured by the paired survival method. Preliminary results obtained with a dose of 5 Gy were consistent with those obtained using 15 Gy. These results suggest the further development of the comet assay for clinical studies. 21 refs., 7 figs., 5 tabs

  19. The Comet Assay and its applications in the field of ecotoxicology: a mature tool that continues to expand its perspectives

    Science.gov (United States)

    de Lapuente, Joaquín; Lourenço, Joana; Mendo, Sónia A.; Borràs, Miquel; Martins, Marta G.; Costa, Pedro M.; Pacheco, Mário

    2015-01-01

    Since Singh and colleagues, in 1988, launched to the scientific community the alkaline Single Cell Gel Electrophoresis (SCGE) protocol, or Comet Assay, its uses and applications has been increasing. The thematic areas of its current employment in the evaluation of genetic toxicity are vast, either in vitro or in vivo, both in the laboratory and in the environment, terrestrial or aquatic. It has been applied to a wide range of experimental models: bacteria, fungi, cells culture, arthropods, fishes, amphibians, reptiles, mammals, and humans. This document is intended to be a comprehensive review of what has been published to date on the field of ecotoxicology, aiming at the following main aspects: (i) to show the most relevant experimental models used as bioindicators both in the laboratory and in the field. Fishes are clearly the most adopted group, reflecting their popularity as bioindicator models, as well as a primary concern over the aquatic environment health. Amphibians are among the most sensitive organisms to environmental changes, mainly due to an early aquatic-dependent development stage and a highly permeable skin. Moreover, in the terrestrial approach, earthworms, plants or mammalians are excellent organisms to be used as experimental models for genotoxic evaluation of pollutants, complex mix of pollutants and chemicals, in both laboratory and natural environment. (ii) To review the development and modifications of the protocols used and the cell types (or tissues) used. The most recent developments concern the adoption of the enzyme linked assay (digestion with lesion-specific repair endonucleases) and prediction of the ability to repair of oxidative DNA damage, which is becoming a widespread approach, albeit challenging. For practical/technical reasons, blood is the most common choice but tissues/cells like gills, sperm cells, early larval stages, coelomocytes, liver or kidney have been also used. (iii) To highlight correlations with other biomarkers

  20. Protective effects of aqueous and ethanolic extracts of Portulaca oleracea L. aerial parts on H2O2-induced DNA damage in lymphocytes by comet assay.

    Science.gov (United States)

    Behravan, Javad; Mosafa, Fatemeh; Soudmand, Negar; Taghiabadi, Elahe; Razavi, Bibi Marjan; Karimi, Gholamreza

    2011-09-01

    The comet assay is a standard method for measuring DNA damage. In this study, the protective effects of ethanolic and aqueous extracts of Portulaca oleracea L. (P. oleracea) on human lymphocyte DNA lesions were evaluated with the comet assay. Lymphocytes were isolated from blood samples taken from healthy volunteers. Human lymphocytes were incubated in H(2)O(2) (50,100, and 200 μM), aqueous extract (0.05, 0.1, 0.5, 1, and 2.5mg/ml), and ethanolic extracts (0.05, 0.1, 0.5, 1, and 2.5mg/ml) of P. oleraceae aerial parts alone with a combination of H(2)O(2) (100 μM) with either 1 or 2.5mg/ml of both extracts at 4°C for 30 minutes. The extent of DNA migration was measured using the alkaline single cell gel electrophoresis approach assay, and DNA damage was expressed as percentage tail DNA. We found that the aqueous extract of P. oleracea significantly inhibited DNA damage, while there was no effect of the ethanolic extract. These data suggest that the aqueous extract of P. oleracea can prevent oxidative DNA damage to human lymphocytes, which is likely due to antioxidant constituents in the extract. Copyright © 2011. Published by Elsevier B.V.

  1. The Comet Assay and its applications in the field of ecotoxicology: a mature tool that continues to expand its perspectives

    Directory of Open Access Journals (Sweden)

    Joaquin ede Lapuente

    2015-06-01

    Full Text Available Since Singh and colleagues, in 1988, launched to the scientific community the alkaline Single Cell Gel Electrophoresis (SCGE protocol, or Comet Assay, its uses and applications has been increasing. The thematic areas of its current employment in the evaluation of genetic toxicity are vast, either in vitro or in vivo, both in the laboratory and in the environment, terrestrial or aquatic. It has been applied to a wide range of experimental models: bacteria, fungi, cells culture, arthropods, fishes, amphibians, reptiles, mammals and humans. This document is intended to be a comprehensive review of what has been published to date on the field of ecotoxicology, aiming at the following main aspects: (i to show the most relevant experimental models used as bioindicators both in the laboratory and in the field. Fishes are clearly the most adopted group, reflecting their popularity as bioindicator models, as well as a primary concern over the aquatic environment health. Amphibians are among the most sensitive organisms to environmental changes, mainly due to an early aquatic-dependent development stage and a highly permeable skin. Moreover, in the terrestrial approach, earthworms, plants or mammalians are excellent organisms to be used as experimental models for genotoxic evaluation of pollutants, complex mix of pollutants and chemicals, in both laboratory and natural environment. (ii To review the development and modifications of the protocols used and the cell types (or tissues used. The most recent developments concern the adoption of the enzyme linked assay (digestion with lesion-specific repair endonucleases and prediction of the ability to repair of oxidative DNA damage, which is becoming a widespread approach, albeit challenging. For practical/technical reasons, blood is the most common choice but tissues/cells like gills, sperm cells, early larval stages, coelomocytes, liver or kidney have been also used. (iii To highlight correlations with other

  2. Assessment of DNA damage in blood lymphocytes of bakery workers by comet assay.

    Science.gov (United States)

    Kianmehr, Mojtaba; Hajavi, Jafar; Gazeri, Javad

    2017-09-01

    The comet assay is widely used in screening and identification of genotoxic effects of different substances on people in either their working or living environment. Exposure to fuel smoke leads to DNA damage and ultimately different types of cancer. Using a comet assay, the present study aimed to assess peripheral blood lymphocyte DNA damage in people working in bakeries using natural gas, kerosene, diesel, or firewood for fuel compared to those in the control group. The subjects of this study were 55 people in total who were divided into four experimental groups, each of which comprised of 11 members (based on the type of fuel used), and one control group comprised of 11 members. Using CometScore, the subjects' peripheral blood lymphocytes were examined for DNA damage. All bakers, that is, experimental subjects, showed significantly greater peripheral blood lymphocyte DNA damage compared to the individuals in the control group. There was greater peripheral blood lymphocyte DNA damage in bakers who had been using firewood for fuel compared to those using other types of fuel to such an extent that tail moments (µm) for firewood-burning bakers was 4.40 ± 1.98 versus 1.35 ± 0.84 for natural gas, 1.85 ± 1.33 for diesel, and 2.19 ± 2.20 for kerosene. The results indicated that burning firewood is the greatest inducer of peripheral blood lymphocytes DNA damage in bakers. Nonetheless, there was no significant difference in peripheral blood lymphocyte DNA damage among diesel and kerosene burning bakers.

  3. Evaluation of Sinos River water genotoxicity using the comet assay in fish.

    Science.gov (United States)

    Scalon, M C S; Rechenmacher, C; Siebel, A M; Kayser, M L; Rodrigues, M T; Maluf, S W; Rodrigues, M A S; Silva, L B

    2010-12-01

    The Sinos River, in southern Brazil, is polluted by industrial discharges and untreated urban wastes. Fish genotoxicity biomarkers are valuable parameters for environmental risk assessment. In this study, we used the comet assay to detect genotoxicity due to multiple sources of pollution in the peripheral blood of a native fish species (Hyphessobrycon luetkenii). In addition, we analysed possible DNA damage from aluminum, lead, chromium, copper, nickel, iron and zinc contamination. Water samples were collected seasonally from three sampling sites and the fish were assessed under laboratory conditions. Water chemical analysis showed an increased level of aluminum and iron in most of the samples at sites 2 and 3, located in the middle and lower river course, respectively. The index of DNA damage assessed by the comet assay demonstrated no significant differences in different seasons or at the different sampling sites, while the frequency of cells with DNA damage was higher in water samples collected at sites 1 and 2 during the spring season. None of the metals studied seems to be associated with the increase in the frequency of cells with DNA damage observed during the spring season. The results of this study indicate that the Sinos River is contaminated with substances that are genotoxic to fish, including the waters near the river spring.

  4. The agricultural impact of pesticides on Physalaemus cuvieri tadpoles (Amphibia: Anura ascertained by comet assay

    Directory of Open Access Journals (Sweden)

    Macks W. Gonçalves

    2017-10-01

    Full Text Available ABSTRACT Amphibians inhabiting agricultural areas are constantly exposed to large amounts of chemicals, which reach the aquatic environment during the rainy season through runoff, drainage, and leaching. We performed a comet assay on the erythrocytes of tadpoles found in the surroundings of agricultural fields (soybean and corn crops, where there is an intense release of several kinds of pesticides in different quantities. We aimed to detect differences in the genotoxic parameters between populations collected from soybeans and cornfields, and between them and tadpoles sampled from non-agricultural areas (control group. Tadpoles collected from ponds located at soybean fields had significantly more DNA damage, followed by tadpoles collected from cornfields. In contrast, animals sampled from non-agricultural areas had the lowest incidence of DNA damage. In addition, we found a negative correlation between the parameters of the comet assay and the area of the ponds surrounding soybean. This correlation indicates a possible dilution effect in the concentration of pesticides. Finally, Physalaemus cuvieri Fitzinger, 1826 seems to be a good bioindicator for detecting the genotoxic effects of field agricultural insecticides; therefore, we suggest that this species should be used in environmental biomonitoring studies, since it is common and abundant where it occurs.

  5. Assessment of genotoxic damage by the comet assay in white storks (Ciconia ciconia) after the Doñana Ecological Disaster.

    Science.gov (United States)

    Pastor, N; López-Lázaro, M; Tella, J L; Baos, R; Hiraldo, F; Cortés, F

    2001-05-01

    Single cell gel electrophoresis, the so-called "Comet" assay, was performed as a genotoxicity test in white storks sampled in an area heavily contaminated after the ecological disaster in south western Spain. This disaster occurred as a consequence of a massive toxic spillage of acid waste rich in heavy metals that impacted on the Doñana National Park. The importance of this protected area as a breeding and wintering site for many endangered bird species makes this analysis of DNA damage of special interest. Our results clearly show that white storks born in the contaminated area 1 year after the toxic spill bear a high burden of genetic damage as compared with control individuals. The possible implications for future survival as well as reproductive rate are discussed.

  6. DNA damage detected by the alkaline comet assay in the liver of mice after oral administration of tetrachloroethylene

    DEFF Research Database (Denmark)

    Cederberg, H.; Henriksson, J.; Binderup, Mona-Lise

    2010-01-01

    Induction of DNA damage in the liver and kidney of male CD1 mice was studied by means of the alkaline Comet assay after oral administration of tetrachloroethylene at the doses of 1000 and 2000 mg/kg/day. A statistically significant dose-related increase in tail intensity was established in hepato......Induction of DNA damage in the liver and kidney of male CD1 mice was studied by means of the alkaline Comet assay after oral administration of tetrachloroethylene at the doses of 1000 and 2000 mg/kg/day. A statistically significant dose-related increase in tail intensity was established...

  7. Photogenotoxicity of skin phototumorigenic fluoroquinolone antibiotics detected using the comet assay.

    Science.gov (United States)

    Reavy, H J; Traynor, N J; Gibbs, N K

    1997-09-01

    The fluoroquinolone (FQ) antibiotics photosensitize human skin to solar UV radiation and are reported to photosensitize tumor formation in mouse skin. As tumor initiation will not occur without genotoxic insult, we examined the potential of ciprofloxacin, lomefloxacin, fleroxacin, BAYy3118 (a recently developed monofluorinated quinolone) and a nalidixic acid to photosensitize DNA damage in V79 hamster fibroblasts in vitro. Cells were exposed to 37.5 kJ/m2 UVA (320-400 nm; glass filtered Sylvania psoralen + UVA (PUVA) tubes; calibrated Waldmann radiometer) at 4 degrees C in the presence of FQ and immediately afterwards embedded in agarose, lysed and placed in an electrophoretic field at pH 12. Under these denaturing conditions, the presence of DNA single-strand breaks (SSB), alkali-labile sites (ALS) and double-strand breaks (DSB) can be visualized as DNA migrating away from the nucleus (characteristic "comet" appearance) after staining with a specific fluorochrome. At FQ concentrations that induced minimal loss of cell viability (neutral red uptake assay) the compounds tested induced comets with a rank order of BAYy3118 > norfloxacin > ciprofloxacin > lomefloxacin > fleroxacin > nalidixic acid. If cells were incubated after treatment for 1 h at 37 degrees C, the comet score decreased, suggesting efficient removal of SSB/ALS/DSB. Addition of the DNA polymerase(alpha) inhibitor, aphidicolin, to cells treated with either ciprofloxacin alone or ciprofloxacin + UVA resulted in an accumulation of SSB due to the endo/exonuclease steps of excision repair. We have demonstrated that the FQ are photogenotoxic in mammalian cells but the FQ-photosensitized SSB are efficiently repaired. Preliminary evidence that ciprofloxacin photosensitizes the formation of DNA lesions warranting excision repair may indicate production of more mutagenic lesions.

  8. Assessment of status of three water bodies in Serbia based on tissue metal and metalloid concentration (ICP-OES) and genotoxicity (comet assay).

    Science.gov (United States)

    Sunjog, Karolina; Kolarević, Stoimir; Kračun-Kolarević, Margareta; Višnjić-Jeftić, Željka; Skorić, Stefan; Gačić, Zoran; Lenhardt, Mirjana; Vasić, Nebojša; Vuković-Gačić, Branka

    2016-06-01

    Metals and metalloids are natural components of the biosphere, which are not produced per se by human beings, but whose form and distribution can be affected by human activities. Like all substances, they are a contaminant if present in excess compared to background levels and/or in a form that would not normally occur in the environment. Samples of liver, gills, gonads and muscle from European chub, Squalius cephalus, were analyzed for Al, As, B, Ba, Cr, Cu, Fe, Hg, Mn, Mo, Sr and Zn using inductively coupled plasma optical emission spectrometry (ICP-OES) to highlight the importance of tissue selection in monitoring research. The comet assay or single cell gel electrophoresis (SCGE) was selected as an in vivo genotoxicity assay, a rapid and sensitive method for measuring genotoxic effects in blood, liver and gills of the European chub. Microscopic images of comets were scored using Comet IV Computer Software (Perceptive Instruments, UK). The objective of our study was to investigate two reservoirs, Zlatar and Garasi, and one river, Pestan by: (i) determining and comparing metal and metalloid concentrations in sediment, water and tissues of European chub: liver, gills, muscle and gonads (ii) comparing these findings with genotoxicity of water expressed through DNA damage of fish tissues. A clear link between the level of metals in water, sediment and tissues and between metal and genotoxicity levels at examined sites was not found. This suggests that other xenobiotics (possibly the organic compounds), contribute to DNA damage. Copyright © 2016 Elsevier Ltd. All rights reserved.

  9. Genotoxic effects of boric acid and borax in zebrafish, Danio rerio using alkaline comet assay.

    Science.gov (United States)

    Gülsoy, Nagihan; Yavas, Cüneyd; Mutlu, Özal

    2015-01-01

    The present study is conducted to determine the potential mechanisms of Boron compounds, boric acid (BA) and borax (BX), on genotoxicity of zebrafish Danio rerio for 24, 48, 72 and 96-hours acute exposure (level:1, 4, 16, 64 mg/l BA and BX) in semi-static bioassay experiment. For that purpose, peripheral erythrocytes were drawn from caudal vein and Comet assay was applied to assess genotoxicity. Acute (96 hours) exposure and high concentrations of boric acid and borax increases % tail DNA and Olive tail moment. Genotoxicity was found for BA as concentration-dependent and BX as concentration and time dependent manner. In general, significant effects (P borax-induced genotoxicity in fish.

  10. Measuring the effects of pesticides on occupationally exposed humans with the comet assay.

    Science.gov (United States)

    Piperakis, S M; Kontogianni, K; Siffel, C; Piperakis, M M

    2006-08-01

    In this study we examine the effects of a mixture of pesticides on occupationally exposed agricultural workers. The study was performed on 149 people, 84 agricultural workers and 65 healthy men from the same area, who served as the control group. The exposed group was divided into a subgroup with 65 individuals moderately exposed (39 men and 26 women) and a highly exposed subgroup consisted of 19 men. The statistical analysis of the comet assay results showed that there were no significant differences in basal DNA damage between pesticide-exposed workers and the control group nor between moderately and highly exposed ones. In addition, exposure of peripheral blood lymphocytes to hydrogen peroxide or gamma-irradiation led to a similar degree of DNA damage and subsequent repair for all the studied populations. Copyright 2006 Wiley Periodicals, Inc.

  11. Biomonitoring with the comet assay of Greek greenhouse workers exposed to pesticides.

    Science.gov (United States)

    Piperakis, S M; Petrakou, E; Tsilimigaki, S; Sagnou, M; Monogiudis, E; Haniotakis, G; Karkaseli, H; Sarikaki, E

    2003-01-01

    The pesticides in use in Greek greenhouses include a number of agents known to be mutagens and carcinogens. In the present study, we evaluated whether occupational exposure of agricultural workers to a complex mixture of pesticides resulted in a significant increase in DNA damage in human peripheral blood lymphocytes (PBLs). A total of 116 healthy individuals were divided into groups based on exposure to pesticides, smoking status, and gender. Alkaline comet assays performed on PBLs from these individuals indicated no statistically significant differences in basal DNA damage between the study groups. In addition, exposure of PBLs to a dose of hydrogen peroxide led to a similar degree of DNA damage and subsequent repair for all the study populations. The results of the study indicate that the agricultural workers who participated in this study had no detectable increase in DNA damage or alteration in the cellular response to DNA damage. Copyright 2003 Wiley-Liss, Inc.

  12. The Comet Assay for the Evaluation of Genotoxic Potential of Landfill Leachate

    Directory of Open Access Journals (Sweden)

    Kamila Widziewicz

    2012-01-01

    Full Text Available Genotoxic assessment of landfill leachate before and after biological treatment was conducted with two human cell lines (Me45 and NHDF and Daphnia magna somatic cells. The alkali version of comet assay was used to examine genotoxicity of leachate by DNA strand breaks analysis and its repair dynamics. The leachate samples were collected from Zabrze landfill, situated in the Upper Silesian Industrial District, Poland. Statistically significant differences (Kruskal-Wallice ANOVA rank model were observed between DNA strand breaks in cells incubated with leachate before and after treatment (P<0.001. Nonparametric Friedman ANOVA confirmed time-reliable and concentration-reliable cells response to leachate concentration. Examinations of chemical properties showed a marked decrease in leachate parameters after treatment which correlate to reduced genotoxicity towards tested cells. Obtained results demonstrate that biological cotreatment of leachate together with municipal wastewater is an efficient method for its genotoxic potential reduction; however, treated leachate still possessed genotoxic character.

  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. 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

  15. Detection of DNA damage by comet fluorescence in situ hybridization.

    Science.gov (United States)

    Schlörmann, Wiebke; Glei, Michael

    2012-01-01

    Comet fluorescence in situ hybridization (Comet-FISH) is a useful method to detect overall and region-specific DNA damage in individual cells. Two well-established methods are combined, the Comet assay (single cell gel electrophoresis) and fluorescence in situ hybridization (FISH). The Comet assay is the method of choice for the detection of DNA damage. With the alkaline version the influence of specific substances such as water pollutants or ingredients of food on individual cells can be easily measured. The Comet assay involves the embedding of cells in agarose on microscopic slides, lysis of cells, and separation of DNA via electrophoresis. Damaged DNA migrates from the nucleus (head of the comet) forming a tail. The percentage of DNA in the tail correlates with the degree of DNA strand breaks (DNA damage). The combination of FISH with the Comet assay uses labeled probes which hybridize specifically to selected DNA sequences. This allows the detection of specific DNA damage or repair capacity in single cells. Here we present exemplarily the Comet-FISH method by detection of DNA damage using hydrogen peroxide as a genotoxic model substrate.

  16. DNA Strand Breaks in Mitotic Germ Cells of Caenorhabditis elegans Evaluated by Comet Assay.

    Science.gov (United States)

    Park, Sojin; Choi, Seoyun; Ahn, Byungchan

    2016-03-01

    DNA damage responses are important for the maintenance of genome stability and the survival of organisms. Such responses are activated in the presence of DNA damage and lead to cell cycle arrest, apoptosis, and DNA repair. In Caenorhabditis elegans, double-strand breaks induced by DNA damaging agents have been detected indirectly by antibodies against DSB recognizing proteins. In this study we used a comet assay to detect DNA strand breaks and to measure the elimination of DNA strand breaks in mitotic germline nuclei of C. elegans. We found that C. elegans brc-1 mutants were more sensitive to ionizing radiation and camptothecin than the N2 wild-type strain and repaired DNA strand breaks less efficiently than N2. This study is the first demonstration of direct measurement of DNA strand breaks in mitotic germline nuclei of C. elegans. This newly developed assay can be applied to detect DNA strand breaks in different C. elegans mutants that are sensitive to DNA damaging agents.

  17. Evaluation the urban atmospheric conditions in different cities using comet and micronuclei assay in Tradescantia pallida.

    Science.gov (United States)

    Sposito, Juliana Caroline Vivian; Crispim, Bruno do Amaral; Romãn, Amanda Izadora; Mussury, Rosilda Mara; Pereira, Joelson Gonçalves; Seno, Leonardo Oliveira; Grisolia, Alexeia Barufatti

    2017-05-01

    In the present study, genotoxicity and mutagenicity were investigated in Tradescantia pallida exposed to vehicular traffic at different sites in a high-altitude tropical climate. During March, May, July, September, and November 2014, a comet assay and micronucleus bioassays were conducted on young inflorescences and leaves of T. pallida collected from twelve towns in the southern region of Mato Grosso do Sul with different amounts of vehicular traffic. Weather parameters (temperature, relative humidity and rainfall) were measured and vehicles were counted to determine traffic levels in each town. A higher frequency of genotoxic and mutagenic damage was observed in the municipality of Dourados. The highest frequency of genetic damage was observed in September and November according to both assays. Relative humidity and rainfall were inversely proportional to the frequency of genetic damage in T. pallida during the collection period. Based on these results, we conclude that the bioassays are efficient for assessing the effects of vehicular traffic in these towns with respect to weather conditions over time. These bioassays can be applied to identify risk areas, which are determined by climatic conditions and air pollutants released. Copyright © 2017 Elsevier Ltd. All rights reserved.

  18. Comparison of DNA comet assay and germination test (half-embryo-test) in gamma-irradiated cherry seeds

    International Nuclear Information System (INIS)

    Todoroki, Setsuko; Hayashi, Toru

    2002-01-01

    Cherry fruits were irradiated with gamma-rays at doses up to 200Gy (effective dose for disinfestation of codling moth), and DNA strand break in seed embryos was investigated by using alkaline comet assay. Immediately after irradiation (≥100Gy), DNA from embryos produced comets with a long and wide tail due to fragmentation. In control cells, DNA relaxed and produced comet with very short tail (with few strand break). After 72h storage, DNA from fruits irradiated at 200 Gy showed comets with little tail and tail moment of comets was same as un-irradiated control. These results indicate that the strand breaks of DNA caused by irradiation in fresh seed embryo are repaired during storage. On the contrary, the ability of germination lost by irradiation did not restored, a dose of 100Gy and more retarded shoot elongation. In cherries irradiated at 100Gy, the shooting percentage was less than 50% at 4th day after incubation. Germination test (Half embryo test) can be discriminate between irradiated and un-irradiated cherries. (author)

  19. Comparison of DNA comet assay and germination test (half-embryo-test) in gamma-irradiated cherry seeds

    International Nuclear Information System (INIS)

    Todoroki, S.; Hayashi, T.

    2002-01-01

    Cherry fruits were irradiated with gamma-rays at doses up to 200Gy (effective dose for disinfestation of codling moth), and DNA strand break in seed embryos was investigated by using alkaline comet assay. Immediately after irradiation (>-100Gy), DNA from embryos produced comets with a long and wide tail due to fragmentation. In control cells, DNA relaxed and produced comet with very short tail (with few strand break). After 72h storage, DNA from fruits irradiated at 200 Gy showed comets with little tail and tail moment of comets was same as un-irradiated control. These results indicate that the strand breaks of DNA caused by irradiation in fresh seed embryo are repaired during storage. On the contrary, the ability of germination lost by irradiation did not restored, a dose of 100Gy and more retarded shoot elongation. In cherries irradiated at 100Gy, the shooting percentage was less than 50% at 4th day after incubation. Germination test (Half embryo test) can be discriminate between irradiated and un-irradiated cherries

  20. Genotoxicity of nano/microparticles in in vitro micronuclei, in vivo comet and mutation assay systems

    Directory of Open Access Journals (Sweden)

    Fukumori Nobutaka

    2009-09-01

    Full Text Available Abstract Background Recently, manufactured nano/microparticles such as fullerenes (C60, carbon black (CB and ceramic fiber are being widely used because of their desirable properties in industrial, medical and cosmetic fields. However, there are few data on these particles in mammalian mutagenesis and carcinogenesis. To examine genotoxic effects by C60, CB and kaolin, an in vitro micronuclei (MN test was conducted with human lung cancer cell line, A549 cells. In addition, DNA damage and mutations were analyzed by in vivo assay systems using male C57BL/6J or gpt delta transgenic mice which were intratracheally instilled with single or multiple doses of 0.2 mg per animal of particles. Results In in vitro genotoxic analysis, increased MN frequencies were observed in A549 cells treated with C60, CB and kaolin in a dose-dependent manner. These three nano/microparticles also induced DNA damage in the lungs of C57BL/6J mice measured by comet assay. Moreover, single or multiple instillations of C60 and kaolin, increased either or both of gpt and Spi- mutant frequencies in the lungs of gpt delta transgenic mice. Mutation spectra analysis showed transversions were predominant, and more than 60% of the base substitutions occurred at G:C base pairs in the gpt genes. The G:C to C:G transversion was commonly increased by these particle instillations. Conclusion Manufactured nano/microparticles, CB, C60 and kaolin, were shown to be genotoxic in in vitro and in vivo assay systems.

  1. Evaluation of DNA Damage in Common Carp (Cyprinus carpio L. by Comet Assay for Determination of Possible Pollution in Lake Mogan (Ankara

    Directory of Open Access Journals (Sweden)

    İsmet Çok

    2011-01-01

    Full Text Available Contamination of the aquatic environment with various concentrations of pollutants results in unexpected threats to humans and wildlife. The consequences of exposure and metabolism of pollutants/xenobiotics, especially carcinogens and mutagens, can be suitably assessed by investigating severe events, such as DNA damage; for example, DNA adducts and DNA strand breaks. One of the commonly used techniques to detect DNA damage in aquatic organisms is single-cell gel electrophoresis (comet assay. This study was carried out using Cyprinus carpio in order to identify the possible pollution in Lake Mogan, near Ankara, Turkey, where the city's sewer system and pesticides used in agriculture are believed to be the common causes of pollution. From the comet assay, the tail length (μm, tail intensity (%, and tail moment values of fish caught from Lake Mogan were found to be 31.10 ± 10.39, 7.77 ± 4.51, 1.50 ± 1.48, respectively, whereas for clean reference sites they were found to be 22.80 ± 1.08, 3.47 ± 1.59, 0.40 ± 0.51, respectively. The values are statistically different from each other (p < 0.0001, p < 0.0001, and p < 0.0013, respectively. These results indicate that Lake Mogan may be polluted with substances that have genotoxic effects and constitute an early warning for the lake system. Further detailed research is needed to establish the source of the pollution and the chemicals responsible.

  2. Assessing genotoxicity of diuron on Drosophila melanogaster by the wing-spot test and the wing imaginal disk comet assay.

    Science.gov (United States)

    Peraza-Vega, Ricardo I; Castañeda-Sortibrán, América N; Valverde, Mahara; Rojas, Emilio; Rodríguez-Arnaiz, Rosario

    2017-05-01

    The aim of this study was to evaluate the genotoxicity of the herbicide diuron in the wing-spot test and a novel wing imaginal disk comet assay in Drosophila melanogaster. The wing-spot test was performed with standard (ST) and high-bioactivation (HB) crosses after providing chronic 48 h treatment to third instar larvae. A positive dose-response effect was observed in both crosses, but statistically reduced spot frequencies were registered for the HB cross compared with the ST. This latter finding suggests that metabolism differences play an important role in the genotoxic effect of diuron. To verify diuron's ability to produce DNA damage, a wing imaginal disk comet assay was performed after providing 24 h diuron treatment to ST and HB third instar larvae. DNA damage induced by the herbicide had a significantly positive dose-response effect even at very low concentrations in both strains. However, as noted for the wing-spot test, a significant difference between strains was not observed that could be related to the duration of exposure between both assays. A positive correlation between the comet assay and the wing-spot test was found with regard to diuron genotoxicity.

  3. The DNA 'comet assay' as a rapid screening technique to control irradiated food.

    Science.gov (United States)

    Cerda, H; Delincée, H; Haine, H; Rupp, H

    1997-04-29

    The exposure of food to ionizing radiation is being progressively used in many countries to inactivate food pathogens, to eradicate pests, and to extend shelf-life, thereby contributing to a safer and more plentiful food supply. To ensure free consumer choice, irradiated food will be labelled as such, and to enforce labelling, analytical methods to detect the irradiation treatment in the food product itself are desirable. In particular, there is a need for simple and rapid screening methods for the control of irradiated food. The DNA comet assay offers great potential as a rapid tool to detect whether a wide variety of foodstuffs have been radiation processed. In order to simplify the test, the agarose single-layer set-up has been chosen, using a neutral protocol. Interlaboratory blind trials have been successfully carried out with a number of food products, both of animal and plant origin. This paper presents an overview of the hitherto obtained results and in addition the results of an intercomparison test with seeds, dried fruits and spices are described. In this intercomparison, an identification rate of 95% was achieved. Thus, using this novel technique, an effective screening of radiation-induced DNA fragmentation is obtained. Since other food treatments also may cause DNA fragmentation, samples with fragmented DNA suspected to have been irradiated should be analyzed by other validated methods for irradiated food, if such treatments which damage DNA cannot be excluded.

  4. The DNA `comet assay` as a rapid screening technique to control irradiated food

    Energy Technology Data Exchange (ETDEWEB)

    Cerda, H. [Department of Radioecology, The Swedish University of Agricultural Sciences, Uppsala (Sweden); Delincee, H. [Institute of Nutritional Physiology, Federal Research Centre for Nutrition, Karlsruhe (Germany); Haine, H. [Campden and Chorleywood Food Research Association, Chipping Campden, Gloucestershire (United Kingdom); Rupp, H. [Swiss Federal Office of Public Health, Section of Food Chemistry, Berne (Switzerland)

    1997-04-29

    The exposure of food to ionizing radiation is being progressively used in many countries to inactivate food pathogens, to eradicate pests, and to extend shelf-life, thereby contributing to a safer and more plentiful food supply. To ensure free consumer choice, irradiated food will be labelled as such, and to enforce labelling, analytical methods to detect the irradiation treatment in the food product itself are desirable. In particular, there is a need for simple and rapid screening methods for the control of irradiated food. The DNA comet assay offers great potential as a rapid tool to detect whether a wide variety of foodstuffs have been radiation processed. In order to simplify the test, the agarose single-layer set-up has been chosen, using a neutral protocol. Interlaboratory blind trials have been successfully carried out with a number of food products, both of animal and plant origin. This paper presents an overview of the hitherto obtained results and in addition the results of an intercomparison test with seeds, dried fruits and spices are described. In this intercomparison, an identification rate of 95% was achieved. Thus, using this novel technique, an effective screening of radiation-induced DNA fragmentation is obtained. Since other food treatments also may cause DNA fragmentation, samples with fragmented DNA suspected to have been irradiated should be analyzed by other validated methods for irradiated food, if such treatments which damage DNA cannot be excluded.

  5. The DNA 'comet assay' as a rapid screening technique to control irradiated food

    International Nuclear Information System (INIS)

    Cerda, H.; Delincee, H.; Haine, H.; Rupp, H.

    1997-01-01

    The exposure of food to ionizing radiation is being progressively used in many countries to inactivate food pathogens, to eradicate pests, and to extend shelf-life, thereby contributing to a safer and more plentiful food supply. To ensure free consumer choice, irradiated food will be labelled as such, and to enforce labelling, analytical methods to detect the irradiation treatment in the food product itself are desirable. In particular, there is a need for simple and rapid screening methods for the control of irradiated food. The DNA comet assay offers great potential as a rapid tool to detect whether a wide variety of foodstuffs have been radiation processed. In order to simplify the test, the agarose single-layer set-up has been chosen, using a neutral protocol. Interlaboratory blind trials have been successfully carried out with a number of food products, both of animal and plant origin. This paper presents an overview of the hitherto obtained results and in addition the results of an intercomparison test with seeds, dried fruits and spices are described. In this intercomparison, an identification rate of 95% was achieved. Thus, using this novel technique, an effective screening of radiation-induced DNA fragmentation is obtained. Since other food treatments also may cause DNA fragmentation, samples with fragmented DNA suspected to have been irradiated should be analyzed by other validated methods for irradiated food, if such treatments which damage DNA cannot be excluded

  6. Radiomodifying effect of resveratrol in human rhabdomyosarcoma (RD) cell culture applying the comet assay

    International Nuclear Information System (INIS)

    Magalhaes, Vanessa D.; Rogero, Sizue O.; Vieira, Daniel P.; Okazaki, Kayo; Rogero, Jose R.; Cruz, Aurea S.

    2013-01-01

    Cancer is considered a worldwide public health problem. Resveratrol is a defense polyphenol, synthesized naturally by a wide variety of plants according to response of ultraviolet radiation (UV) exposition or according to mechanical stress resulting of pathogens or chemical and physical agents. In vines this substance is found in elevated concentration. Thus, resveratrol is present in grape juice and wines, especially red wine. Red wines are the best dietary source of resveratrol.The protective effects performed by resveratrol during the process of cell damage, produced by oxidative effects of free radicals, are anti-inflammatory, anti-platelet and anti-carcinogenic activity, prevent or inhibit degenerative diseases, decrease incidence of cardiovascular diseases. Moreover, resveratrol is considered as a cell radioprotector. On the other hand, in some elevated concentrations resveratrol is considered as a radiosensitizing compound. The aim of this work was study in vitro the radiomodifying effect of resveratrol in human rhabdomyosarcoma (RD) cells applying the comet assay to evaluate the cellular damage and its repair capacity. In this study RD cells culture was irradiated by gamma radiation at 50 Gy and 100 Gy doses and the used resveratrol concentrations was from 15 μM to 60 μM. The protective and radioprotective effects were observed at 15 μM and 30 μM resveratrol concentrations. The resveratrol concentration of 60 μM showed cytotoxic effect to RD tumor cells and with gamma radiation presence this concentration showed no statistically significant radiosensitizing effects. (author)

  7. DNA damage levels in electronics workers in Southern China: A micro-whole blood comet assay.

    Science.gov (United States)

    Zhao, Zhiqiang; Xing, Xiumei; Ou, Xiaoyan; Liu, Xinxia; Zhou, Ridong; Zhang, Huimin; Yang, Linqing; Zhuang, Zhixiong; Su, Xiaolin; Lu, Yao; Jiang, Jun; Yang, Yarui; Cui, Dong; He, Yun

    2017-10-01

    We evaluated DNA damage levels of different categories of workers exposed to hazards inside electronics factories in Southern China. To find out the most dangerous risk factor, a cross-sectional study was conducted on a total of 584 exposed subjects and 138 controls in an electronics factory in Southern China, where the electronics industry is prevalent. The exposed hazards included isopropanol (IPO), lead, noise, video display terminals (VDT), lead in a high-temperature (high-temp) environment, and IPO in a high-temp environment. DNA damage detection was performed by the micro-whole blood comet assay using peripheral blood. DNA damage levels were estimated by percent tail DNA (%T). Linear regression models were used to test DNA damage differences between exposed groups and control group with adjustments for potential confounding factors. The level of DNA damage was more significant in both lead in a high-temp and IPO in a high-temp environment groups than in that of the controls (pIPO, lead, noise, VDT environment and controls. In conclusion, we identified potential risk factors for DNA damage to electronics workers. Special attention should be paid to workers exposed to IPO and lead in a high-temp environment. Copyright © 2017 Elsevier B.V. All rights reserved.

  8. Radiomodifying effect of resveratrol in human rhabdomyosarcoma (RD) cell culture applying the comet assay

    Energy Technology Data Exchange (ETDEWEB)

    Magalhaes, Vanessa D.; Rogero, Sizue O.; Vieira, Daniel P.; Okazaki, Kayo; Rogero, Jose R., E-mail: van.biologa@gmail.com [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil); Cruz, Aurea S., E-mail: aurcruz@ial.sp.gov.br [Instituto Adolfo Lutz (IAL-SP), Sao Paulo, SP (Brazil)

    2013-07-01

    Cancer is considered a worldwide public health problem. Resveratrol is a defense polyphenol, synthesized naturally by a wide variety of plants according to response of ultraviolet radiation (UV) exposition or according to mechanical stress resulting of pathogens or chemical and physical agents. In vines this substance is found in elevated concentration. Thus, resveratrol is present in grape juice and wines, especially red wine. Red wines are the best dietary source of resveratrol.The protective effects performed by resveratrol during the process of cell damage, produced by oxidative effects of free radicals, are anti-inflammatory, anti-platelet and anti-carcinogenic activity, prevent or inhibit degenerative diseases, decrease incidence of cardiovascular diseases. Moreover, resveratrol is considered as a cell radioprotector. On the other hand, in some elevated concentrations resveratrol is considered as a radiosensitizing compound. The aim of this work was study in vitro the radiomodifying effect of resveratrol in human rhabdomyosarcoma (RD) cells applying the comet assay to evaluate the cellular damage and its repair capacity. In this study RD cells culture was irradiated by gamma radiation at 50 Gy and 100 Gy doses and the used resveratrol concentrations was from 15 μM to 60 μM. The protective and radioprotective effects were observed at 15 μM and 30 μM resveratrol concentrations. The resveratrol concentration of 60 μM showed cytotoxic effect to RD tumor cells and with gamma radiation presence this concentration showed no statistically significant radiosensitizing effects. (author)

  9. Comet assay comparison of different Corbicula fluminea (Mollusca tissues for the detection of genotoxicity

    Directory of Open Access Journals (Sweden)

    Janaina Rigonato

    2005-09-01

    Full Text Available The comet assay was used to study the sensitivity of the widely distributed freshwater bivalve mollusk Corbicula fluminea to the DNA-damaging alkylating-agent methylmethane sulfonate (MMS. This study was undertaken to ascertain if C. fluminea is a good bioindicator of pollutants in aquatic environments and identify which C. fluminea tissue is most effective and practical for genotoxicity studies. The mollusks were exposed to 0.6, 1.2 or 2.4 X 10-4 M MMS for 40 min and their hemolymph, gill tissue and digestive gland tissue assessed for the level of DNA damage and the time needed for the tissues to recovery. Regression analysis showed a direct linear dose-response relationship between MMS concentration and the number of damaged cells for hemolymph and digestive gland tissue but a quadratic relationship for gill tissue, which made the interpretation the gill tissue results difficult. The basal level of DNA damage to gill tissue was very high, possibly because gill is the organs most directly exposed to environmental toxins and mutagenic agents. Although all three types of tissue produced useful results, hemolymph and digestive gland tissue produced more reproducible and reliable results. Hemolymph was the best sample type in that it was easy to obtain and handle, while gill tissue required more manipulation to obtain cell suspensions. Our results indicate that C. fluminea is an optimal bioindicator for the determination genotoxic contaminants in aquatic environments.

  10. Genotoxicity of oxy-PAHs to Japanese medaka (Oryzias latipes) embryos assessed using the comet assay.

    Science.gov (United States)

    Dasgupta, Subham; Cao, Austin; Mauer, Brittany; Yan, Beizhan; Uno, Seiichi; McElroy, Anne

    2014-12-01

    Polycyclic aromatic hydrocarbons (PAHs) have long been recognized as important environmental toxicants. Despite a plethora of information on the fate and effects of parent PAHs, relatively little is known about the environmental fate and toxicity of ketone- and quinone-substituted PAH oxidation products (termed oxy-PAHs), particularly in the aquatic environment. This study begins to fill that gap using embryos of the Japanese medaka (Oryzias latipes) as a model species. The genotoxic potential of two environmentally relevant oxy-PAHs, acenaphthenequinone and 7,12-benz[a]anthracenquinone, was assessed using the comet assay. We found that both oxy-PAHs could cause significant increases in DNA damage after only 48 h of exposure at the lowest concentrations tested (5 μg/L). Comparisons of the genotoxic potential between these oxy-PAHs and their corresponding parent PAHs (acenaphthene and benz[a]anthracene) and a well-known mutagenic PAH, benzo[a]pyrene, indicated similar potencies among all five of these compounds, particularly after longer (7 day) exposures. This study demonstrates the mutagenic potential of oxy-PAHs to an in vivo fish embryo model and points out the need for further study of their environmental occurrence and biologic effects.

  11. Comet assay in the detection of irradiated garlic; Teste do cometa na deteccao de alho irradiado

    Energy Technology Data Exchange (ETDEWEB)

    Villavicencio, Anna Lucia C.H.; Marin-Huachaca, Nelida Simona; Romanelli, Maria Fernanda [Instituto de Pesquisas Energeticas e Nucleares (IPEN), Sao Paulo, SP (Brazil)]. E-mail: villavic@net.ipen.br; Delincee, Henry [Federal Research Centre for Nutrition - BFE, Karlsruhe (Germany)]. E-mail: henry.delincee@bfe.uni-karlsruhe.de

    2002-07-01

    The increased claim for fresh produce has forced a consensus between nations to pay more attention to the phytosanitary regulations. Inhibition of sprouting of bulbs and tubers by applying ionising radiation is authorised by the National Food Codes in Brazil. The availability of methods for detection of irradiated food will contribute to increase consumers' confidence. A quick and simple screening test to indicate whether a food product has been irradiated or not was utilised in this study. The DNA comet assay was applied to verify whether garlic imported from China had been irradiated or not. This test has already been adopted as a European Standard (EN 13784), for detection of irradiated food. Non-irradiated control samples of garlic and garlic treated with maleic hydrazide were compared with garlic samples irradiated in our department. The unirradiated samples exhibited only limited DNA migration. If samples were irradiated, an increased DNA fragmentation was observed which permitted the discrimination between non-irradiated and irradiated samples. Since the garlic samples from China showed only very limited DNA fragmentation, they were deemed non-irradiated. Thus, this simple screening test was shown to be successful for identification of an irradiation treatment. (author)

  12. DNA comet assay to identify different freezing temperatures of irradiated liver chicken

    International Nuclear Information System (INIS)

    Duarte, Renato C.; Mozeika, Michel A.; Fanaro, Gustavo B.; Villavicencio, Anna L.C.H.; Marchioni, Eric

    2009-01-01

    The cold chain is a succession of steps which maintain the food at low temperature. The thawed food never be frozen again and the best solution being to consume it quickly to avoid the microorganism growth which causes decay and nutrients damage. One of most important point is that freezing process, unlike irradiation, do not destroy microorganisms, only inactive them as long as they remain in a frozen state. The Comet Assay is an original test used to detect irradiated foods that's recognize the DNA damage and can then be used to control the overall degradation of the food and in a certain extend to evaluate the damage caused by irradiation, different forms of freeze and storage time on liver chicken cells. Different freezing temperatures were used, deep freeze -196 deg C and slow freeze -10 deg C. Samples were irradiated in a 60 Co irradiator with 1.5, 3.0 and 4.5 kGy radiation doses. Fast freezing technique induces a low percent of DNA degradation comparing to slow freezing technique. This procedure could be a good choose to chicken freezing processing. (author)

  13. New comprehensive denaturing-gradient-gel-electrophoresis assay for KRAS mutation detection applied to paraffin-embedded tumours

    NARCIS (Netherlands)

    Hayes, VM; Westra, JL; Verlind, E; Bleeker, W; Plukker, JT; Hofstra, RMW; Buys, CHCM

    2000-01-01

    A comprehensive mutation detection assay is presented for the entire coding region and all splice site junctions of the KRAS oncogene. The assay is based on denaturing gradient gel electrophoresis and applicable to archival paraffin-embedded tumour material. All KRAS amplicons are analysed within

  14. 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

  15. The impact of lymphocyte isolation on induced DNA damage in human blood samples measured by the comet assay.

    Science.gov (United States)

    Bausinger, Julia; Speit, Günter

    2016-09-01

    The comet assay is frequently used in human biomonitoring for the detection of exposure to genotoxic agents. Peripheral blood samples are most frequently used and tested either as whole blood or after isolation of lymphocytes (i.e. peripheral blood mononuclear cells, PBMC). To investigate a potential impact of lymphocyte isolation on induced DNA damage in human blood samples, we exposed blood ex vivo to mutagens with different modes of genotoxic action. The comet assay was performed either directly with whole blood at the end of the exposure period or with lymphocytes isolated directly after exposure. In addition to the recommended standard protocol for lymphocyte isolation, a shortened protocol was established to optimise the isolation procedure. The results indicate that the effects of induced DNA strand breaks and alkali-labile sites induced by ionising radiation and alkylants, respectively, are significantly reduced in isolated lymphocytes. In contrast, oxidative DNA base damage (induced by potassium bromate) and stable bulky adducts (induced by benzo[a]pyrene-7,8-dihydrodiol-9,10-epoxide; BPDE) seem to be less affected. Our findings suggest that in vivo-induced DNA damage might also be reduced in isolated lymphocytes in comparison with the whole blood depending of the types of DNA damage induced. Because only small genotoxic effects can generally be expected in human biomonitoring studies with the comet assay after occupational and environmental exposure to genotoxic agents, any loss might be relevant and should be avoided. The possibility of such effects and their potential impact on variability of comet assay results in human biomonitoring should be considered when performing or evaluating such kind of studies. © The Author 2016. Published by Oxford University Press on behalf of the UK Environmental Mutagen Society. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  16. Development of cultures of the marine sponge Hymeniacidon perleve for genotoxicity assessment using the alkaline comet assay.

    Science.gov (United States)

    Akpiri, Rachael U; Konya, Roseline S; Hodges, Nikolas J

    2017-12-01

    Sponges are a potential alternative model species to bivalves in pollution biomonitoring and environmental risk assessment in the aquatic ecosystem. In the present study, a novel in vivo exposure sponge culture model was developed from field-collected and cryopreserved sponge (Hymeniacidon perleve) cells to investigate the genotoxic effects of environmentally relevant metals in the laboratory. Sponge cell aggregates were cultured and exposed to noncytotoxic concentrations (0-0.4 mg/L) of cadmium chloride, nickel chloride, and sodium dichromate as quantified by the reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and DNA-strand breaks assessed by the comet assay. Reactive oxygen species (ROS) formation was quantified by oxidation of 2',7'-dichlorofluorescin diacetate in sponge cell aggregates exposed to the same concentrations of Cd, Cr, and Ni. There was a statistically significant (p metals investigated. To the best of our knowledge, we have utilized for the first time the alkaline comet assay to detect DNA-strand breaks in marine sponge cells and demonstrated that exposure to noncytotoxic concentrations of Cd, Cr, and Ni for 12 h results in a concentration-dependent increase in DNA damage and levels of ROS production. In conclusion, we have developed a novel in vivo model based on culture of cryopreserved sponge cells that is compatible with the alkaline comet assay. Genotoxicity in marine sponges measured by the comet assay technique may be a useful tool for biomonitoring research and risk assessment in aquatic ecosystems. Environ Toxicol Chem 2017;36:3314-3323. © 2017 SETAC. © 2017 SETAC.

  17. Evaluation of γ-radiation-induced DNA damage in two species of bivalves and their relative sensitivity using comet assay

    Energy Technology Data Exchange (ETDEWEB)

    Praveen Kumar, M.K., E-mail: here.praveen@gmail.com [Department of Zoology, Goa University, Goa 403206 (India); Shyama, S.K., E-mail: skshyama@gmail.com [Department of Zoology, Goa University, Goa 403206 (India); Sonaye, B.S. [Department of Radiation Oncology, Goa Medical College, Goa (India); Naik, U Roshini; Kadam, S.B.; Bipin, P.D.; D’costa, A. [Department of Zoology, Goa University, Goa 403206 (India); Chaubey, R.C. [Radiation Biology and Health Science Division, Bhabha Atomic Research Centre, Mumbai (India)

    2014-05-01

    Highlights: • Possible genotoxic effect of accidental exposure of aquatic fauna to γ radiation. • Relative sensitivity of bivalves to γ radiation is also analyzed using comet assay. • γ radiation induced significant genetic damage in both the species of bivalves. • P. malabarica and M. casta exhibited a similar level of sensitivity to γ radiation. • Comet assay may be used as a biomarker for the environmental biomonitoring. - Abstract: Ionizing radiation is known to induce genetic damage in diverse groups of organisms. Under accidental situations, large quantities of radioactive elements get released into the environment and radiation emitted from these radionuclides may adversely affect both the man and the non-human biota. The present study is aimed (a) to know the genotoxic effect of gamma radiation on aquatic fauna employing two species of selected bivalves, (b) to evaluate the possible use of ‘Comet assay’ for detecting genetic damage in haemocytes of bivalves as a biomarker for environmental biomonitoring and also (c) to compare the relative sensitivity of two species of bivalves viz. Paphia malabarica and Meretrix casta to gamma radiation. The comet assays was optimized and validated using different concentrations (18, 32 and 56 mg/L) of ethyl methanesulfonate (EMS), a direct-acting reference genotoxic agent, to which the bivalves were exposed for various times (24, 48 and 72 h). Bivalves were irradiated (single acute exposure) with 5 different doses (viz. 2, 4, 6, 8 and 10 Gy) of gamma radiation and their genotoxic effects on the haemocytes were studied using the comet assay. Haemolymph was collected from the adductor muscle at 24, 48 and 72 h of both EMS-exposed and irradiated bivalves and comet assay was carried out using standard protocol. A significant increase in DNA damage was observed as indicated by an increase in % tail DNA damage at different concentrations of EMS and all the doses of gamma radiation as compared to controls in

  18. Evaluation of γ-radiation-induced DNA damage in two species of bivalves and their relative sensitivity using comet assay

    International Nuclear Information System (INIS)

    Praveen Kumar, M.K.; Shyama, S.K.; Sonaye, B.S.; Naik, U Roshini; Kadam, S.B.; Bipin, P.D.; D’costa, A.; Chaubey, R.C.

    2014-01-01

    Highlights: • Possible genotoxic effect of accidental exposure of aquatic fauna to γ radiation. • Relative sensitivity of bivalves to γ radiation is also analyzed using comet assay. • γ radiation induced significant genetic damage in both the species of bivalves. • P. malabarica and M. casta exhibited a similar level of sensitivity to γ radiation. • Comet assay may be used as a biomarker for the environmental biomonitoring. - Abstract: Ionizing radiation is known to induce genetic damage in diverse groups of organisms. Under accidental situations, large quantities of radioactive elements get released into the environment and radiation emitted from these radionuclides may adversely affect both the man and the non-human biota. The present study is aimed (a) to know the genotoxic effect of gamma radiation on aquatic fauna employing two species of selected bivalves, (b) to evaluate the possible use of ‘Comet assay’ for detecting genetic damage in haemocytes of bivalves as a biomarker for environmental biomonitoring and also (c) to compare the relative sensitivity of two species of bivalves viz. Paphia malabarica and Meretrix casta to gamma radiation. The comet assays was optimized and validated using different concentrations (18, 32 and 56 mg/L) of ethyl methanesulfonate (EMS), a direct-acting reference genotoxic agent, to which the bivalves were exposed for various times (24, 48 and 72 h). Bivalves were irradiated (single acute exposure) with 5 different doses (viz. 2, 4, 6, 8 and 10 Gy) of gamma radiation and their genotoxic effects on the haemocytes were studied using the comet assay. Haemolymph was collected from the adductor muscle at 24, 48 and 72 h of both EMS-exposed and irradiated bivalves and comet assay was carried out using standard protocol. A significant increase in DNA damage was observed as indicated by an increase in % tail DNA damage at different concentrations of EMS and all the doses of gamma radiation as compared to controls in

  19. Photons bring light into DNA repair: the comet assay and laser microbeams for studying photogenotoxicity of drugs and ageing.

    Science.gov (United States)

    Greulich, Karl Otto

    2011-03-01

    This contribution reviews recent applications of micromanipulation, by UV photons, in DNA repair and ageing research as well as in the evaluation of the phototoxicity of drugs. In some cases, micromanipulation is combined with the comet assay, a technique, which allows a direct view on DNA damages. It is shown that, in humans, the sensitivity of DNA to UV induced damage and its subsequent repair is surprisingly stable up to high age and that drugs which are usually non-toxic induce DNA damage when irradiated in parallel by UV irradiation. Using the immune fluorescent comet assay, IFCA, a variant of the comet assay, direct comparison of the effects of ionizing (137) Cs radiation with those of localized UV radiation is possible. When a laser microbeam is used to damage DNA in a cell nucleus with high temporal and spatial resolution, it can be observed directly how repair molecules accumulate (are recruited) at the site of damage. Comparison of the recruitment speed allows establishing an order of DNA repair events. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  20. Factors influencing heterogeneity of radiation-induced DNA-damage measured by the alkaline comet assay

    International Nuclear Information System (INIS)

    Seidel, Clemens; Lautenschläger, Christine; Dunst, Jürgen; Müller, Arndt-Christian

    2012-01-01

    To investigate whether different conditions of DNA structure and radiation treatment could modify heterogeneity of response. Additionally to study variance as a potential parameter of heterogeneity for radiosensitivity testing. Two-hundred leukocytes per sample of healthy donors were split into four groups. I: Intact chromatin structure; II: Nucleoids of histone-depleted DNA; III: Nucleoids of histone-depleted DNA with 90 mM DMSO as antioxidant. Response to single (I-III) and twice (IV) irradiation with 4 Gy and repair kinetics were evaluated using %Tail-DNA. Heterogeneity of DNA damage was determined by calculation of variance of DNA-damage (V) and mean variance (Mvar), mutual comparisons were done by one-way analysis of variance (ANOVA). Heterogeneity of initial DNA-damage (I, 0 min repair) increased without histones (II). Absence of histones was balanced by addition of antioxidants (III). Repair reduced heterogeneity of all samples (with and without irradiation). However double irradiation plus repair led to a higher level of heterogeneity distinguishable from single irradiation and repair in intact cells. Increase of mean DNA damage was associated with a similarly elevated variance of DNA damage (r = +0.88). Heterogeneity of DNA-damage can be modified by histone level, antioxidant concentration, repair and radiation dose and was positively correlated with DNA damage. Experimental conditions might be optimized by reducing scatter of comet assay data by repair and antioxidants, potentially allowing better discrimination of small differences. Amount of heterogeneity measured by variance might be an additional useful parameter to characterize radiosensitivity

  1. Improved single-cell gel electrophoresis assay for detecting DNA damage in Eisenia foetida.

    Science.gov (United States)

    Di Marzio, Walter D; Saenz, María E; Lemière, Sebastien; Vasseur, Paule

    2005-12-01

    The earthworm (Eisenia foetida) is an attractive sentinel species for detecting genotoxicity in soil. In this study, an improved single-cell gel electrophoresis (SCGE) assay was developed for detecting DNA damage in the coelomocytes (lymphocytes) of earthworms. Coelomocytes were obtained from the coelomic fluid using a modified extrusion medium that did not include the mucolytic agent guaiacol. The extruded coelomocytes contained at least three types of cells: eleocytes (75% of the total), amoebocytes, and granulocytes. The DNA migration parameters were determined for untreated cells of each type in order that the assay could be performed with minimum inter- and intra-individual variation. In addition, lysis time was reduced to 10 min, and only one neutralization step was used. DNA damage was detected in isolated eleocytes treated with hydrogen peroxide and cadmium, and in eleocytes from earthworms exposed for up to 21 days to soil containing polycyclic aromatic hydrocarbons. The SCGE assay using earthworm eleocytes appears to be a sensitive biomarker for evaluating exposure to genotoxic compounds.

  2. The alkaline comet assay as a biomarker of primary DNA damage in peripheral blood leukocytes of nuclear medicine personnel

    International Nuclear Information System (INIS)

    Kopjar, N.; Garaj-Vrhovac, V.

    2003-01-01

    The aim of this study was to assess whether occupational exposure to chronic low doses of ionizing radiation in nuclear medicine departments may lead to genotoxicity. The alkaline comet assay was selected as a bio-marker of exposure to evaluate the levels of primary DNA damage in peripheral blood leukocytes of exposed and corresponding control subjects. Statistically significant differences were found between comet tail length and tail moment values measured in leukocytes from the exposed and control groups. Within exposed group significant inter-individual differences in DNA damage were assessed, indicating different genome sensitivity. In majority of exposed subjects the levels of DNA damage were in positive correlation with the duration of occupational exposure, while the influences of age and dosimeter readings could be excluded. However, the levels of primary DNA damage detected both in control and exposed subjects were significantly influenced by smoking. The present study indicates the possibility of genotoxic risks related to occupational exposure in nuclear medicine departments. Therefore, the exposed personnel should carefully apply the radiation protection procedures to minimize, as low as possible, radiation exposure to avoid possible genotoxic effects. According to results obtained, the alkaline comet assay could be usefully applied as a sensitive additional bio-marker in the regular health screening of workers occupationally exposed to low doses of ionizing radiation. (authors)

  3. Construction of dose-effect curves by irradiating peripheral blood mononuclear cells analyzed with the alkaline comet assay

    International Nuclear Information System (INIS)

    Rueda, Lenin; Rosales Jaime; Leon, Kety; Espinoza, Marco

    2015-01-01

    The suitability of the alkaline comet assay to detect levels of DNA damage in human monocytes and lymphocytes under radiation was tested. For this purpose, four dose-effect curves by irradiation of cells isolated from blood samples of 4 young, health and non-smoker donors. A gamma ray source of 60 Co was used and doses of 0, 2.5, 5 and 7 Gy were delivered to blood cells. The results show good correlation (R2 = 0.973, SD = 0.009) between the radiation dose and DNA damage as measured by the parameter Tail Intensity (percentage of DNA in the tail of the 'comet'). Furthermore, we observe the variability in predicting DNA damage by Gy (7.59, SD = 1.5). Potential use of this technique in the biological dosimetry of ionizing radiations is being evaluated. (author)

  4. Comparative evaluation of the in vitro micronucleus test and the alkaline single cell gel electrophoresis assay for the detection of DNA damaging agents: genotoxic effects of cobalt powder, tungsten carbide and cobalt-tungsten carbide.

    Science.gov (United States)

    Van Goethem, F; Lison, D; Kirsch-Volders, M

    1997-08-01

    Although it is well known that micronuclei may arise from either DNA breakage leading to acentric chromosome fragments or from chromosome/chromatid lagging in anaphase, the ratio between the amount of DNA breakage induced and the frequency of micronuclei expressed in the following interphase is unclear. With the development of the alkaline single cell gel electrophoresis assay, which measures single strand and/or double strand breaks in a cell by cell approach, it is new possible to address this question at the cellular level. We therefore compared the genotoxic potential of pure cobalt powder (Co) and a cobalt-containing alloy, cobalt-tungsten carbide (WC-Co), involved in specific lung disorders, in parallel with the alkaline single cell gel electrophoresis (SCGE) assay (comet assay) and the cytokinesis-blocked micronucleus (MN) test, both carried out in vitro on isolated human leukocytes. The comet assay indicated that the WC-Co mixture produced a higher level of DNA damage than Co alone; WC alone was not able to induce a dose-dependent DNA breakage effect as was seen for Co and WC-Co. Results from the MN test confirmed these observations. It was clear that the clastogenic property of Co-containing dust is significantly enhanced when the Co metal is mixed with WC and suggested that their physicochemical characteristics may act as one of the important parameters responsible for the increased incidence of lung cancers observed in the population of hard metal workers. In agreement with data obtained in the same laboratory on liposoluble chemicals (PCBs and chlorinated aliphatic hydrocarbons) and from the literature, the results indicate that both the comet assay and the micronucleus test were able to detect differences in the genotoxic potential of the compounds studied. Although the micronucleus test seemed to be less sensitive to assess a synergistic DNA damaging potential of the mixture involved, it detects chromosomal aberrations (chromosome/genome mutations

  5. Hepatocytes, rather than leukocytes reverse DNA damage in vivo induced by whole body y-irradiation of mice, as shown by the alkaline comet assay

    Directory of Open Access Journals (Sweden)

    JUANA PINCHEIRA

    2008-01-01

    Full Text Available DNA damage repair was assessed in quiescent (G0 leukocytes and in hepatocytes of mice, after 1 and 2 hours recovery from a single whole body y-irradiation with 0.5, 1 or 2 Gy. Evaluation of single-strand breaks (SSB and alkali-labile sites together were carried out by a single-cell electrophoresis at pH>13.0 (alkaline comet assay. In non-irradiated (control mice, the constitutive, endogenous DNA damage (basal was around 1.5 times higher in leukocytes than in hepatocytes. Irradiation immediately increased SSB frequency in both cell types, in a dose-dependent manner. Two sequential phases took place during the in vivo repair of the radio-induced DNA lesions. The earliest one, present in both hepatocytes and leukocytes, further increased the SSB frequency, making evident the processing of some primary lesions in DNA bases into the SSB repair intermediates. In a second phase, SSB frequency decreased because of their removal. In hepatocytes, such a frequency regressed to the constitutive basal level after 2 hours recovery from either 0.5 orí Gy. On the other hand, the SSB repair phase was specifically abrogated in leukocytes, at the doses and recovery times analyzed. Thus, the efficiency of in vivo repair of radio-induced DNA damage in dormant cells (lymphocytes is quite different from that in hepatocytes whose low proliferation activity accounts only for cell renewal.

  6. Usefulness of capillary electrophoresis-based multiplex PCR assay for species-specific identification of Candida spp.

    Science.gov (United States)

    Mallus, F; Martis, S; Serra, C; Loi, G; Camboni, T; Manzin, A

    2013-02-15

    The study evaluated the performances of a commercial multiplex PCR assay, the Seegene Seeplex STI Master Panel 3, for Candida spp. identification. Eighty clinical strains of Candida spp. were identified with this system and a homemade multiplex PCR assay. The results were also compared with those obtained with two phenotypic methods. The study provided a preliminary evaluation of a multiplex assay from Seegene that uses capillary electrophoresis as the detection of amplified products. The Seeplex assay was found to be a rapid and useful method for identifying large numbers of yeast isolates in the clinical laboratory context. Copyright © 2012 Elsevier B.V. All rights reserved.

  7. The alkaline comet assay used in evaluation of genotoxic damage of drinking water disinfection by-products (bromoform and chloroform

    Directory of Open Access Journals (Sweden)

    Messaouda Khallef

    2015-06-01

    Full Text Available The alkaline comet assay (pH 12.3 is a useful method for monitoring genotoxic effects of environmental pollutants in the root nuclei of Allium cepa and various plants; it allows the detection of single- and double-strand breaks, incomplete excision-repair sites and cross-links. It has been introduced to detect even small changes in DNA structure. It is a technically simple, highly sensitive, fast and economic test which detects in vitro and in vivo genotoxicity (DNA integrity and packing mode in any cell types examined, and requires just a few cells for its execution (Liman et al., 2011; Yıldız et al., 2009. Chloroform and bromoform are the most important trihalomethanes found in drinking water. Different concentrations of bromoform (25, 50, 75and 100µg/ml and chloroform (25, 50, 100 and 200 µg/ml were introduced to onion tuber roots. Distilled water was used as a negative control and methyl methansulfonate (MMS-10 µg/ml as positive control. All obtained data were subjected to statistical analyses by using SPSS 15.0 for Windows software. For comparison purposes, Duncan multiple range tests using one-way analysis of variance (ANOVA were employed and p<0.05 was accepted as the test of significance. Comet assay results showed that DNA damage was significant at p <0.05 for the different concentrations of chloroform and bromoform compared to the negative control which has a damage rate equal to 3.5 ± 0.7 and the positive control which has damage rate equal to 13.5 ± 2.12. The exposure of root tip cells to these disinfection by-products increases DNA damage. All concentrations examined in this study of bromoform and chloroform cause significant harm, which could be due to DNA damage induced by oxidative stress. The measurement of DNA damage in the nuclei of higher plant tissues is a new area of study with SCGE. This assay could be incorporated into in situ monitoring of atmosphere, water and soil: the comet assay allows a fast detection without

  8. Evaluation of sperm DNA quality in men presenting with testicular cancer and lymphoma using alkaline and neutral Comet assays.

    Science.gov (United States)

    Kumar, K; Lewis, S; Vinci, S; Riera-Escamilla, A; Fino, M-G; Tamburrino, L; Muratori, M; Larsen, P; Krausz, C

    2018-01-01

    Despite more cancers in young men over the past two decades, improvements in therapies give a greater chance to live full lives following treatment. Sperm genomic quality is variable following cancer diagnosis, so its assessment is important if sperm cryopreservation is being considered. Here, we evaluated DNA damage using two DNA damage assays: an alkaline and for the first time, a neutral Comet assays in men presenting with testicular cancer (n = 19 for alkaline and 13 for neutral group) and lymphoma (n = 13 for alkaline and 09 for neutral group) compared with fertile donors (n = 20 for alkaline and 14 for neutral group). No significant differences were observed in any semen analysis parameters. In contrast, sperm DNA damage was higher in men with testicular cancer than in donors as assessed by both the alkaline (12.4% vs. 37.4%, p alkaline (35.0% vs. 12.4%) and neutral (10.7% against 7.5% (p alkaline and neutral Comet assays is more sensitive than semen analysis in detecting impaired sperm quality in men presenting with cancer. It may provide a useful adjunct when considering storage prior to cancer investigations and assisted reproductive techniques (ART)-based treatment. © 2017 American Society of Andrology and European Academy of Andrology.

  9. The comet assay as a rapid test in biomonitoring occupational exposure to DNA-damaging agents and effect of confounding factors

    DEFF Research Database (Denmark)

    Møller, P; Knudsen, Lisbeth E.; Loft, S

    2000-01-01

    Within the last decade, the comet assay has been used with increasing popularity to investigate the level of DNA damage in terms of strand breaks and alkaline labile sites in biomonitoring studies. The assay is easily performed on WBCs and has been included in a wide range of biomonitoring studies...... appeared to have less power than the positive studies. Also, there were poor dose-response relationships in many of the biomonitoring studies. Many factors have been reported to produce effects by the comet assay, e.g., age, air pollution exposure, diet, exercise, gender, infection, residential radon...... be used as criteria for the selection of populations and that data on exercise, diet, and recent infections be registered before blood sampling. Samples from exposed and unexposed populations should be collected at the same time to avoid seasonal variation. In general, the comet assay is considered...

  10. Effect of chronic low dose natural radiation in human peripheral blood mononuclear cells: Evaluation of DNA damage and repair using the alkaline comet assay

    Energy Technology Data Exchange (ETDEWEB)

    Kumar, P.R. Vivek, E-mail: prvkumar06@gmail.com [Low Level Radiation Research Laboratory, Radiation Biology and Health Sciences Division, Bio-Science Group, Bhabha Atomic Research Centre, IRE Campus, Beach Road, Kollam 691 001, Kerala (India); Seshadri, M. [Low Level Radiation Research Section, Radiation Biology and Health Sciences Division, Bio-Science Group, Bhabha Atomic Research Centre, Trombay, Mumbai 400 085 (India); Jaikrishan, G. [Low Level Radiation Research Laboratory, Radiation Biology and Health Sciences Division, Bio-Science Group, Bhabha Atomic Research Centre, IRE Campus, Beach Road, Kollam 691 001, Kerala (India); Das, Birajalaxmi [Low Level Radiation Research Section, Radiation Biology and Health Sciences Division, Bio-Science Group, Bhabha Atomic Research Centre, Trombay, Mumbai 400 085 (India)

    2015-05-15

    Highlights: • Effect of chronic low dose natural radiation in radio adaptive response studied. • PBMCs of subjects from NLNRA and HLNRA were challenged with gamma radiation. • DNA damage and repair in PBMCs was compared using the alkaline comet assay. • Significant reduction in DNA damage in subjects of high dose group from HLNRA noted. • Probable induction of an in vivo radio adaptive response in subjects from HLNRA. - Abstract: This study investigates whether peripheral blood mononuclear cells (PBMCs) from inhabitants of Kerala in southwest India, exposed to chronic low dose natural radiation in vivo (>1 mSv year{sup −1}), respond with a radioadaptive response to a challenging dose of gamma radiation. Toward this goal, PBMCs isolated from 77 subjects from high-level natural radiation areas (HLNRA) and 37 subjects from a nearby normal level natural radiation area (NLNRA) were challenged with 2 Gy and 4 Gy gamma radiation. Subjects from HLNRA were classified based on the mean annual effective dose received, into low dose group (LDG) and high dose group (HDG) with mean annual effective doses of 2.69 mSv (N = 43, range 1.07 mSv year{sup −1} to 5.55 mSv year{sup −1}) and 9.62 mSv (N = 34, range 6.07 mSv year{sup −1} to17.41 mSv year{sup −1}), respectively. DNA strand breaks and repair kinetics (at 7 min, 15 min and 30 min after 4 Gy) were evaluated using the alkaline single cell gel electrophoresis (comet) assay. Initial levels of DNA strand breaks observed after either a 2 Gy or a 4 Gy challenging dose were significantly lower in subjects of the HDG from HLNRA compared to subjects of NLNRA (2 Gy, P = 0.01; 4 Gy, P = 0.02) and LDG (2 Gy P = 0.01; 4 Gy, P = 0.05). Subjects of HDG from HLNRA showed enhanced rejoining of DNA strand breaks (HDG/NLNRA, P = 0.06) during the early stage of repair (within 7 min). However at later times a similar rate of rejoining of strand breaks was observed across the groups (HDG, LDG and NLNRA). Preliminary results from

  11. Aptamer-based microchip electrophoresis assays for amplification detection of carcinoembryonic antigen.

    Science.gov (United States)

    Pan, Li; Zhao, Jingjin; Huang, Yong; Zhao, Shulin; Liu, Yi-Ming

    2015-10-23

    Carcinoembryonic antigen (CEA) as one of the most widely used tumor markers is used in the clinical diagnosis of colorectal, pancreatic, gastric, and cervical carcinomas. We developed an aptamer-based microchip electrophoresis assay technique for assaying CEA in human serum for cancer diagnosis. The magnetic beads (MBs) are employed as carriers of double strand DNA that is formed by an aptamer of the target and a complementary DNA of the aptamer. After the aptamer in the MB-dsDNA conjugate binds with the target, the complementary DNA was released from the MB-dsDNA conjugate. The released complementary DNA hybridizes with a fluorescein amidite (FAM) labeled DNA, and forms a DNA duplex, which triggers the selective cleavage of FAM labeled DNA by nicking endonuclease Nb.BbvCI, and generating a FAM labeled DNA segment. The released complementary DNA hybridizes with another FAM labeled DNA, resulting in a continuous cleavage of FAM labeled DNA, and the generation of large numbers of FAM labeled DNA segments. In MCE laser induced fluorescence detection (LIF), the FAM labeled DNA segment is separated and detected. The linear range for CEA was 130 pg/ml-8.0 ng/ml with a correlation coefficient of 0.9916 and a detection limit of 68 pg/ml. The CEA concentration in the serum samples from healthy subjects was found to be in the range 1.3 ng/ml to 3.2 ng/ml. The CEA concentration in the samples from cancer patients was found to be >15 ng/ml. This method may become a useful tool for rapid analysis of CEA and other tumor markers in biomedical analysis and clinical diagnosis. Copyright © 2015 Elsevier B.V. All rights reserved.

  12. Genetic Alterations in Pesticide Exposed Bolivian Farmers: An evaluation by analysis of chromosomal aberrations and the comet assay.

    Science.gov (United States)

    Jørs, Erik; Gonzáles, Ana Rosa; Ascarrunz, Maria Eugenia; Tirado, Noemi; Takahashi, Catharina; Lafuente, Erika; Dos Santos, Raquel A; Bailon, Natalia; Cervantes, Rafael; O, Huici; Bælum, Jesper; Lander, Flemming

    2007-11-12

    Pesticides are of concern in Bolivia because of increasing use. Frequent intoxications have been demonstrated due to use of very toxic pesticides, insufficient control of distribution and sale and little knowledge among farmers of protective measures and hygienic procedures. Questionnaires were applied and blood tests taken from 81 volunteers from La Paz County, of whom 48 were pesticide exposed farmers and 33 non-exposed controls. Sixty males and 21 females participated with a mean age of 37.3 years (range 17-76). Data of exposure and possible genetic damage were collected and evaluated by well known statistical methods, controlling for relevant confounders. To measure genetic damage chromosomal aberrations and the comet assay analysis were performed. Pesticide exposed farmers had a higher degree of genetic damage compared to the control group. The number of chromosomal aberrations increased with the intensity of pesticide exposure. Females had a lower number of chromosomal aberrations than males, and people living at altitudes above 2500 metres seemed to exhibit more DNA damage measured by the comet assay. Bolivian farmers showed signs of genotoxic damage, probably related to exposure to pesticides. Due to the potentially negative long term health effects of genetic damage on reproduction and the development of cancer, preventive measures are recommended. Effective control with imports and sales, banning of the most toxic pesticides, education and information are possible measures, which could help preventing the negative effects of pesticides on human health and the environment.

  13. Genetic Alterations in Pesticide Exposed Bolivian Farmers: An evaluation by analysis of chromosomal aberrations and the comet assay

    Directory of Open Access Journals (Sweden)

    Erik Jørs

    2007-01-01

    Full Text Available Background: Pesticides are of concern in Bolivia because of increasing use. Frequent intoxications have been demonstrated due to use of very toxic pesticides, insufficient control of distribution and sale and little knowledge among farmers of protective measures and hygienic procedures.Method: Questionnaires were applied and blood tests taken from 81 volunteers from La Paz County, of whom 48 were pesticide exposed farmers and 33 non-exposed controls. Sixty males and 21 females participated with a mean age of 37.3 years (range 17–76. Data of exposure and possible genetic damage were collected and evaluated by well known statistical methods, controlling for relevant confounders. To measure genetic damage chromosomal aberrations and the comet assay analysis were performed.Results: Pesticide exposed farmers had a higher degree of genetic damage compared to the control group. The number of chromosomal aberrations increased with the intensity of pesticide exposure. Females had a lower number of chromosomal aberrations than males, and people living at altitudes above 2500 metres seemed to exhibit more DNA damage measured by the comet assay.Conclusions: Bolivian farmers showed signs of genotoxic damage, probably related to exposure to pesticides. Due to the potentially negative long term health effects of genetic damage on reproduction and the development of cancer, preventive measures are recommended. Effective control with imports and sales, banning of the most toxic pesticides, education and information are possible measures, which could help preventing the negative effects of pesticides on human health and the environment.

  14. [Comet assay of DNA fragmentation: modification of silver staining for obtaining permanent preparations].

    Science.gov (United States)

    Kamins'kyĭ, V O; Lutsyk, M D; Stoĭka, R S

    2005-01-01

    Modification of comet analysis is proposed for obtaining permanent preparations by DNA staining with silver compounds. The sensitivity of staining is similar to that observed at the treatment by ethidium bromide and other fluorochromes. The advantages of the method are stability of slides and possibility of their reinvestigation by light microscopy. The method does not need expensive fluorescent microscope and lacks contacting with carcinogenic compounds and UV light irradiation.

  15. Comparison of the Freelite serum free light chain (SFLC) assay with serum and urine electrophoresis/immunofixation and the N Latex FLC assay.

    Science.gov (United States)

    Sasson, S C; McGill, K; Wienholt, L; Carr, A; Brown, D A; Kelleher, A D; Breit, S N; Sewell, W A

    2015-10-01

    Few reports have compared available serum free light chain (SFLC) assays. Here, a retrospective audit of the Freelite SFLC assay compared results to electrophoresis (EP)/immunofixation (IFX) and the N Latex FLC assay.A total of 244 samples collected over 3.5 months were studied using the Freelite and N Latex FLC nephelometry assays. Results were compared with serum and/or urine EP/IFX. The precision and linearity of the N Latex FLC assay was examined.Detectable paraprotein by serum or urine EP/IFX was present in 94% of samples with kappa and 100% with lambda FLC restriction. The correlation between the assays was higher for kappa (rho = 0.97) than lambda (rho = 0.89) especially when lambda results were above the upper limit of normal (rho = 0.62). Agreement in the categorical diagnosis as measured by the Cohen's kappa statistic was good (0.70). The N Latex FLC assay displayed good precision and linearity. In discordant samples the Freelite and N Latex FLC assays had equivalent agreement with IFX.Traditional methods of EP/IFX detected paraproteins in the majority of cases. Correlation between the Freelite and N Latex FLC assay is better for kappa than lambda FLC. The two assays are not entirely equivalent. Care should be taken by interpreting physicians and laboratories considering switching assays.

  16. DNA single- and double-strand breaks by alkaline- and immuno-comet assay in lymphocytes of workers exposed to styrene.

    Science.gov (United States)

    Fracasso, Maria Enrica; Doria, Denise; Carrieri, Mariella; Bartolucci, Giovanni Battista; Quintavalle, Sonia; De Rosa, Edoardo

    2009-02-25

    Occupational exposure to styrene was studied in 34 workers employed in the production of fiberglass-reinforced plastic sheets and compared to 29 unexposed healthy controls. We evaluated genotoxic effects induced by occupational styrene exposure in lymphocytes by alkaline version of the comet assay to detect single-strand breaks (SSBs), DNA oxidation products (formamido pyrimidine glycosilase (Fpg)- and endonuclease (Endo III)-sensitive sites) and DNA repair kinetics studies, as well as the neutral version of comet assay for DNA double-strand breaks (DSBs). An innovative aspect of this study was the use of immuno-comet assay, a new technique that recognizes DSBs with specific antibody by DAPI/FITC method. The battery of parameters included markers of external and internal exposure. Exposed workers showed significant high levels of SSBs (p<0.0001) and DSBs (p<0.0001) in neutral- and immuno-comet assay. A drastic decrease in DNA repair activity as compared to controls was observed (180 min vs. 35 min). Styrene workplace concentration significantly correlated with alkaline comet parameters (TM, p=0.013; TI, p=0.008), in negative with TL (p=0.022), and with DNA-base oxidation (TM Endo III, p=0.048 and TI Endo III, p=0.028). There was a significant negative correlation between urinary metabolites (MA+PGA) and TM Endo III (p=0.032) and TI Endo III (p=0.017).

  17. Double Stranded Sperm DNA Breaks, Measured by Comet Assay, Are Associated with Unexplained Recurrent Miscarriage in Couples without a Female Factor

    Science.gov (United States)

    Ribas-Maynou, Jordi; García-Peiró, Agustín; Fernandez-Encinas, Alba; Amengual, Maria José; Prada, Elena; Cortés, Pilar; Navarro, Joaquima; Benet, Jordi

    2012-01-01

    It is known that sperm samples from recurrent pregnancy loss (RPL) couples have an increase in their sperm DNA fragmentation (SDF), but no studies have been performed in order to identify differences between single stranded SDF (ssSDF) and double stranded SDF (dsSDF) in these patients. This could be relevant because the type of DNA damage could have different effects. Semen samples were classified attending their clinical status: 25 fertile donors and 20 RPL patients with at least two unexplained first trimester miscarriages. SDF was analysed using alkaline and neutral Comet assay, SCD test and pulsed-field gel electrophoresis (PFGE), and ROC analysis including data from 105 more infertile patients (n = 150) was performed to establish predictive threshold values. SDF for alkaline and neutral Comet, and the SCD test was analysed in these categories of individuals. Data revealed the presence of two subgroups within fertile donors. The values obtained were 21.10±9.13, 23.35±10.45 and 12.31±4.31, respectively, for fertile donors with low values for both ssSDF and dsSDF; 27.86±12.64, 80.69±12.67 and 12.43±5.22, for fertile donors with low ssSDF and high dsSDF; and 33.61±15.50, 84.64±11.28 and 19.28±6.05, for unexplained RPL patients, also showing a low ssSDF and high dsSDF profile. This latter profile was seen in 85% of unexplained RPL and 33% of fertile donors, suggesting that it may be associated to a male risk factor for undergoing RPL. ROC analysis regarding recurrent miscarriage set the cut-off value at 77.50% of dsDNA SDF. PFGE for low ssSDF and high dsSDF profile samples and positive controls treated with DNase, to induce dsDNA breaks, showed a more intense band of about 48 kb, which fits the toroid model of DNA compaction in sperm, pointing out that some nuclease activity may be affecting their sperm DNA in RPL patients. This work identifies a very specific SDF profile related to the paternal risk of having RPL. PMID:23028579

  18. Assessment of genotoxicity and depuration of anthracene in the juvenile coastal fish Trachinotus carolinus using the comet assay

    Directory of Open Access Journals (Sweden)

    Fabio Matsu Hasue

    2013-12-01

    Full Text Available In the environment, anthracene is characterized as being persistent, bioaccumulative and toxic to aquatic organisms. Biotransformation of xenobiotic substances, such as anthracene, produces reactive oxygen species that may induce DNA strand breaks. The aim of the present study was to evaluate the DNA damage in juvenile T. carolinus exposed to different concentrations (8, 16 and 32 µg.L-1 of anthracene for 24 h in the dark then subsequently allowed to depurate in clean water for different periods of time (48, 96 or 144 h using the comet assay. Our results show that anthracene is genotoxic to T. carolinus and that DNA damage was dose- and depuration/time- dependent. Anthracenegenotoxicity was observed in all experimental concentrations. Depuration seemed to be more efficient in fish exposed to thelowest anthracene concentration and maintained in clean water for 96 h.

  19. Evaluation of the genotoxic potential of the Casearia sylvestris extract on HTC and V79 cells by the comet assay.

    Science.gov (United States)

    Maistro, E L; Carvalho, J C T; Mantovani, M S

    2004-06-01

    Casearia sylvestris is common in tropical America growing wild in Brazil in the states of Amazonas and São Paulo. Its leaves are used in Brazilian folk medicine for several diseases. The present investigation was carried out to examine the genotoxic effects of a C. sylvestris crude ethanolic extract on Hepatoma Tissue Culture (HTC cells) of Rattus norvegicus and Chinese hamster V79 cells in culture, using the comet assay. For the genotoxic evaluation the cells were treated with three different concentrations (0.5, 1 and 2 mg/ml) of extract prepared from a 25 mg/ml aqueous solution. The positive control was cyclophosphamide for HTC cells and methyl methanesulfonate for V79 cells. The duration of the treatment was 2 h. The results showed that the extract of C. sylvestris presented no genotoxic effects and not modified effect inducing DNA damage by alkylating agents cyclophosphamide and methyl methanesulfonate in HTC and V 79 cells respectively.

  20. The use of comet assay in measuring DNA damage and repair efficiency in child, adult, and old age populations.

    Science.gov (United States)

    Piperakis, S M; Kontogianni, K; Karanastasi, G; Iakovidou-Kritsi, Z; Piperakis, M M

    2009-02-01

    In the present study, we used the Comet assay to estimate basal DNA damage in three distinct populations aged 5-10, 40-50, and 60-70 years old. The DNA damage induced by hydrogen peroxide and gamma-irradiation in the lymphocytes of these populations, as well as their repair activity, was also studied. Finally, we measured apoptosis and necrosis after the effect of these agents. Our results indicate that the older population (60-70 years old) showed higher basal levels of DNA damage and was more sensitive to the effects of the DNA-damaging agents than the adult one (40-50 years old), who, in turn, was more sensitive than the younger population (5-10 years old). A decline of the repair efficiency with age to the DNA damage induced by the two agents was also observed. Apoptosis and necrosis were also affected by age.

  1. Mutagenicity and antimutagenicity of (−-hinokinin a trypanosomicidal compound measured by Salmonella microsome and comet assays

    Directory of Open Access Journals (Sweden)

    Resende Flávia Aparecida

    2012-10-01

    Full Text Available Abstract Background The dibenzylbutyrolactone lignan (−-hinokinin (HK was derived by partial synthesis from (−-cubebin, isolated from the dry seeds of the pepper, Piper cubeba. Considering the good trypanosomicidal activity of HK and recalling that natural products are promising starting points for the discovery of novel potentially therapeutic agents, the aim of the present study was to investigate the (anti mutagenic∕ genotoxic activities of HK. Methods The mutagenic∕ genotoxic activities were evaluated by the Ames test on Salmonella typhimurium strains TA98, TA97a, TA100 and TA102, and the comet assay, so as to assess the safe use of HK in the treatment of Chagas’ disease. The antimutagenic ∕antigenotoxic potential of HK were also tested against the mutagenicity of a variety of direct and indirect acting mutagens, such as 4- nitro-o-phenylenediamine (NOPD, sodium azide (SA, mitomycin C (MMC, benzo[a]pyrene (B[a]P, aflatoxin B1 (AFB1, 2-aminoanthracene (2-AA and 2-aminofluorene (2-AF, by the Ames test, and doxorubicin (DXR by the comet assay. Results The mutagenicity∕genotoxicity tests showed that HK did not induce any increase in the number of revertants or extent of DNA damage, demonstrating the absence of mutagenic and genotoxic activities. On the other hand, the results on the antimutagenic potential of HK showed a strong inhibitory effect against some direct and indirect-acting mutagens. Conclusions Regarding the use of HK as an antichagasic drug, the absence of mutagenic effects in animal cell and bacterial systems is encouraging. In addition, HK may be a new potential antigenotoxic ∕ antimutagenic agent from natural sources. However, the protective activity of HK is not general and varies with the type of DNA damage-inducing agent used.

  2. 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.

  3. Monitoring of DNA breakage in embryonic stages of the African catfish Clarias gariepinus (Burchell, 1822) after exposure to lead nitrate using alkaline comet assay

    NARCIS (Netherlands)

    Osman, A.G.M.; Mekkawy, Imam A.; Verreth, J.A.J.; Wuertz, S.; Kloas, W.; Kirschbaum, Frank

    2008-01-01

    Increasing lead contamination in Egyptian ecosystems and high lead concentrations in food items have raised concern for human health and stimulated studies on monitoring ecotoxicological impact of lead-caused genotoxicity. In this work, the alkaline comet assay was modified for monitoring DNA strand

  4. Measurement of DNA base and nucleotide excision repair activities in mammalian cells and tissues using the comet assay - A methodological overview

    Czech Academy of Sciences Publication Activity Database

    Azqueta, A.; Langie, S. A. S.; Slyšková, Jana; Collins, A. R.

    2013-01-01

    Roč. 12, č. 11 (2013), s. 1007-1010 ISSN 1568-7864 Grant - others:EU FP6(XE) LSHB-CT-2006-037575 Institutional support: RVO:68378041 Keywords : comet assay * base excision repair * nucleotide excision repair Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.362, year: 2013

  5. Genotoxic Potential of Two Herbicides and their Active Ingredients Assessed with Comet Assay on a Fish Cell Line, Epithelioma Papillosum Cyprini (EPC)

    DEFF Research Database (Denmark)

    Syberg, Kristian; Rank, Jette; Jensen, Klara

    2013-01-01

    The aim of this study was to optimize the epithelioma papillosum cyprini (EPC) cell line handling procedure for the comet assay to investigate the genotoxic potential of widely used pesticides. The effects of various media and handling of the EPC cell line were examined. Results indicated...

  6. Evaluation of drinking water treatment combined filter backwash water recycling technology based on comet and micronucleus assay.

    Science.gov (United States)

    Chen, Ting; Xu, Yongpeng; Liu, Zhiquan; Zhu, Shijun; Shi, Wenxin; Cui, Fuyi

    2016-04-01

    Based on the fact that recycling of combined filter backwash water (CFBW) directly to drinking water treatment plants (WTP) is considered to be a feasible method to enhance pollutant removal efficiency, we were motivated to evaluate the genotoxicity of water samples from two pilot-scale drinking water treatment systems, one with recycling of combined backwash water, the other one with a conventional process. An integrated approach of the comet and micronucleus (MN) assays was used with zebrafish (Danio rerio) to investigate the water genotoxicity in this study. The total organic carbon (TOC), dissolved organic carbon (DOC), and trihalomethane formation potential (THMFP), of the recycling process were lower than that of the conventional process. All the results showed that there was no statistically significant difference (P>0.05) between the conventional and recycling processes, and indicated that the genotoxicity of water samples from the recycling process did not accumulate in 15 day continuous recycling trial. It was worth noting that there was correlation between the concentrations of TOC, DOC, UV254, and THMFPs in water and the DNA damage score, with corresponding R(2) values of 0.68, 0.63, 0.28, and 0.64. Nevertheless, both DNA strand breaks and MN frequency of all water samples after disinfection were higher than that of water samples from the two treatment units, which meant that the disinfection by-products (DBPs) formed by disinfection could increase the DNA damage. Both the comet and MN tests suggest that the recycling process did not increase the genotoxicity risk, compared to the traditional process. Copyright © 2015. Published by Elsevier B.V.

  7. Genotoxicity assessment of biphasic calcium phosphate of modified porosity on human dental pulp cells using Ames and Comet assays.

    Science.gov (United States)

    Wahab, Nur Fathin Alia Che; Kannan, Thirumulu Ponnuraj; Mahmood, Zuliani; Rahman, Ismail Ab; Ismail, Hanafi

    2018-03-01

    Biphasic Calcium Phosphate (BCP) with a ratio of 20/80 Hydroxyapatite (HA)/Beta-tricalcium phosphate (β-TCP) promotes the differentiation of human dental pulp cells (HDPCs). In the current study, the genotoxicity of locally produced BCP of modified porosity (65%) with a mean pore size of 300micrometer (μm) was assessed using Comet and Ames assays. HDPCs were treated with BCP extract at three different inhibitory concentrations which were obtained based on cytotoxicity test conducted with concurrent negative and positive controls. The tail moment of HDPCs treated with BCP extract at all three concentrations showed no significant difference compared to negative control (p>0.05), indicating that BCP did not induce DNA damage to HDPCs. The BCP was evaluated using five tester strains of Salmonella typhimurium TA98, TA100, TA102, TA1537 and TA1538. Each strain was incubated with BCP extract with five different concentrations in the presence and absence of metabolic activation system (S9) mix. Concurrently, negative and positive controls were included. The average number of revertant colonies per plate treated with the BCP extract was less than double as compared to the number of revertant colonies in negative control plate and no dose-related increase was observed. Results from both assays suggested that the BCP of modified porosity did not exhibit any genotoxic effect under the present test conditions. Copyright © 2017 Elsevier Ltd. All rights reserved.

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

    Science.gov (United States)

    Hussain, Bilal; Sultana, Tayyaba; Sultana, Salma; Masoud, Muhammad Shahreef; Ahmed, Zubair; Mahboob, Shahid

    2018-02-01

    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 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.

  9. Induction and repair of DNA cross-links induced by sulfur mustard in the A-549 cell line followed by a comet assay.

    Science.gov (United States)

    Jost, Petr; Svobodova, Hana; Stetina, Rudolf

    2015-07-25

    Sulfur mustard is a highly toxic chemical warfare agent with devastating impact on intoxicated tissues. DNA cross-links are probably the most toxic DNA lesions induced in the cell by sulfur mustard. The comet assay is a very sensitive method for measuring DNA damage. In the present study using the A-549 lung cell line, the comet assay protocol was optimized for indirect detection of DNA cross-links induced by sulfur mustard. The method is based on the additional treatment of the assayed cells containing cross-links with the chemical mutagen, styrene oxide. Alkali-labile adducts of styrene oxide cause DNA breaks leading to the formation of comets. A significant dose-dependent reduction of DNA migration of the comet's tail was found after exposing cells to sulfur mustard, indicative of the amount of sulfur mustard induced cross-links. The remarkable decrease of % tail DNA could be observed as early as 5min following exposure to sulfur mustard and the maximal effect was found after 30min, when DNA migration was reduced to the minimum. Sulfur mustard preincubated in culture medium without cells lost its ability to induce cross-links and had a half-life of about 15min. Pre-incubation longer than 30min does not lead to a significant increase in cross-links when applied to cells. However, the amount of cross-links is decreased during further incubation due to repair. The current modification of the comet assay provides a useful tool for detecting DNA cross-links induced by sulfur mustard and could be used for detection of other DNA cross-linking agents such as chemotherapeutic drugs. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  10. Application of the microbiological method DEFT/APC and DNA comet assay to detect ionizing radiation processing of minimally processed vegetables

    International Nuclear Information System (INIS)

    Araujo, Michel Mozeika

    2008-01-01

    Marketing of minimally processed vegetables (MPV) are gaining impetus due to its convenience, freshness and apparent healthy. However, minimal processing does not reduce pathogenic microorganisms to safe levels. Food irradiation is used to extend the shelf life and inactivation of food-borne pathogens, Its combination with minimal processing could improve the safety and quality of MPV. Two different food irradiation detection methods, a biological, the DEFT/APC, and another biochemical, the DNA Comet Assay were applied to MPV in order to test its applicability to detect irradiation treatment. DEFT/APC is a microbiological screening method based on the use of the direct epi fluorescent filter technique (DEFT) and the aerobic plate count (APC). DNA Comet Assay detects DNA damage due to ionizing radiation. Samples of lettuce, chard, watercress, dandelion, kale, chicory, spinach, cabbage from retail market were irradiated O.5 kGy and 1.0 kGy using a 60 Co facility. Irradiation treatment guaranteed at least 2 log cycle reduction for aerobic and psychotropic microorganisms. In general, with increasing radiation doses, DEFT counts remained similar independent of irradiation processing while APC counts decreased gradually. The difference of the two counts gradually increased with dose increment in all samples. It could be suggested that a DEFT/APC difference over 2.0 log would be a criteria to judge if a MPV was treated by irradiation. DNA Comet Assay allowed distinguishing non-irradiated samples from irradiated ones, which showed different types of comets owing to DNA fragmentation. Both DEFT/APC method and DNA Comet Assay would be satisfactorily used as a screening method for indicating irradiation processing. (author)

  11. In vivo assessment of DNA damage and protective effects of extracts from Miconia species using the comet assay and micronucleus test.

    Science.gov (United States)

    Serpeloni, Juliana Mara; Bisarro dos Reis, Mariana; Rodrigues, Juliana; Campaner dos Santos, Lourdes; Vilegas, Wagner; Varanda, Eliana A; Dokkedal, Anne L; Cólus, Ilce Mara S

    2008-11-01

    The genus Miconia comprises approximately 1000 species belonging to the Melastomataceae family. Several crude plant extracts from Miconia and their isolated compounds have shown biological activities, such as analgesic and anti-neoplastic action; however, no studies concerning their effects on DNA are available. The present study aimed to evaluate, in vivo, the genotoxic and mutagenic effects of four species of plants from Miconia genus using the comet assay and micronucleus test. Their possible protective effects were also evaluated in experiments associating the plant extracts with cyclophosphamide (CPA). The methanolic extracts of Miconia albicans, Miconia cabucu, Miconia rubiginosa, Miconia stenostachya and the chloroformic extract of M. albicans were investigated. For genotoxic and mutagenic evaluations, three concentrations were tested, 200, 400 and 540 mg/kg body weight (bw), based on the solubility limit of the extract in distilled water. For the protective effects, only the highest dose was evaluated against 40 mg/kg bw of CPA. Blood was removed from mice tails pre- (T0) and post-treatment (T1-30 h) for the micronucleus test and 24 h post-treatment for the comet assay. The Student's t-test was used to compare data obtained at T0 and T1, the analysis of variance-Tukey test was used to compare between groups in the micronucleus test and the Kruskal-Wallis and Dunn's test were used to compare different groups in the comet assay. All the extracts induced alterations in DNA migration (comet assay); however, no mutagenic effect was observed in the micronucleus assay. All extracts showed a protective effect against CPA in both assays. Our study showed that the use of crude extracts could be more advantageous than the use of isolated compounds. The interaction between phytochemicals in the extracts showed efficacy in reducing mutagenicity and improving the protective effects.

  12. Isolation of acetylated bile acids from the sponge Siphonochalina fortis and DNA damage evaluation by the comet assay.

    Science.gov (United States)

    Patiño Cano, Laura P; Bartolotta, Susana A; Casanova, Natalia A; Siless, Gastón E; Portmann, Erika; Schejter, Laura; Palermo, Jorge A; Carballo, Marta A

    2013-10-01

    From the organic extracts of the sponge Siphonochalina fortis, collected at Bahía Bustamante, Chubut, Argentina, three major compounds were isolated and identified as deoxycholic acid 3, 12-diacetate (1), cholic acid 3, 7, 12-triacetate (2) and cholic acid, 3, 7, 12-triacetate. (3). This is the first report of acetylated bile acids in sponges and the first isolation of compound 3 as a natural product. The potential induction of DNA lesions by the isolated compounds was investigated using the comet assay in lymphocytes of human peripheral blood as in vitro model. The results showed that the administration of the bile acid derivatives would not induce DNA damages, indicating that acetylated bile acids are nontoxic metabolites at the tested concentrations. Since the free bile acids were not detected, it is unlikely that the acetylated compounds may be part of the sponge cells detoxification mechanisms. These results may suggest a possible role of acetylated bile acids as a chemical defense mechanism, product of a symbiotic relationship with microorganisms, which would explain their seasonal and geographical variation, and their influence on the previously observed genotoxicity of the organic extract of S. fortis. Copyright © 2013 Elsevier Inc. All rights reserved.

  13. Bioanalysis for biocatalysis: multiplexed capillary electrophoresis-mass spectrometry assay for aminotransferase substrate discovery and specificity profiling.

    Science.gov (United States)

    Mironov, Gleb G; St-Jacques, Antony D; Mungham, Alexander; Eason, Matthew G; Chica, Roberto A; Berezovski, Maxim V

    2013-09-18

    In this work, we introduce an entirely automated enzyme assay based on capillary electrophoresis coupled to electrospray ionization mass spectrometry termed MINISEP-MS for multiple interfluent nanoinjections-incubation-separation-enzyme profiling using mass spectrometry. MINISEP-MS requires only nanoliters of reagent solutions and uses the separation capillary as a microreactor, allowing multiple substrates to be assayed simultaneously. The method can be used to rapidly profile the substrate specificity of any enzyme and to measure steady-state kinetics in an automated fashion. We used the MINISEP-MS assay to profile the substrate specificity of three aminotransferases (E. coli aspartate aminotransferase, E. coli branched-chain amino acid aminotransferase, and Bacillus sp. YM-1 D-amino acid aminotransferase) for 33 potential amino acid substrates and to measure steady-state kinetics. Using MINISEP-MS, we were able to recapitulate the known substrate specificities and to discover new amino acid substrates for these industrially relevant enzymes. Additionally, we were able to measure the apparent K(M) and k(cat) parameters for amino acid donor substrates of these aminotransferases. Because of its many advantages, the MINISEP-MS assay has the potential of becoming a useful tool for researchers aiming to identify or create novel enzymes for specific biocatalytic applications.

  14. Differences in quantification of DNA double-strand breaks assessed by 53BP1/γH2AX focus formation assays and the comet assay in mammalian cells treated with irradiation and N-acetyl-L-cysteine

    International Nuclear Information System (INIS)

    Kurashige, Tomomi; Shimamura, Mika; Nagayama, Yuji

    2016-01-01

    The biological effect of ionizing radiation (IR) on genomic DNA is thought to be either direct or indirect; the latter is mediated by IR induction of free radicals and reactive oxygen species (ROS). This study was designed to evaluate the effect of N-acetyl-L-cysteine (NAC), a well-known ROS-scavenging antioxidant, on IR induction of genotoxicity, cytotoxicity and ROS production in mammalian cells, and aimed to clarify the conflicting data in previous publications. Although we clearly demonstrate the beneficial effect of NAC on IR-induced genotoxicity and cytotoxicity (determined using the micronucleus assay and cell viability/clonogenic assays), the data on NAC's effect on DNA double-strand break (DSB) formation were inconsistent in different assays. Specifically, mitigation of IR-induced DSBs by NAC was readily detected by the neutral comet assay, but not by the γH2AX or 53BP1 focus assays. NAC is a glutathione precursor and exerts its effect after conversion to glutathione, and presumably it has its own biological activity. Assuming that the focus assay reflects the biological responses to DSBs (detection and repair), while the comet assay reflects the physical status of genomic DNA, our results indicate that the comet assay could readily detect the antioxidant effect of NAC on DSB formation. However, NAC's biological effect might affect the detection of DSB repair by the focus assays. Our data illustrate that multiple parameters should be carefully used to analyze DNA damage when studying potential candidates for radioprotective compounds

  15. 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)

    Marcos Vinícius M. Ferraro

    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.

  16. Capillary electrophoresis with electrochemiluminescent detection for highly sensitive assay of genetically modified organisms.

    Science.gov (United States)

    Guo, Longhua; Yang, Huanghao; Qiu, Bin; Xiao, Xueyang; Xue, Linlin; Kim, Donghwan; Chen, Guonan

    2009-12-01

    A capillary electrophoresis coupled with electrochemiluminescent detection system (CE-ECL) was developed for the detection of polymerase chain reaction (PCR) amplicons. The ECL luminophore, tris(1,10-phenanthroline) ruthenium(II) (Ru(phen)(3)(2+)), was labeled to the PCR primers before amplification. Ru(phen)(3)(2+) was then introduced to PCR amplicons by PCR amplification. Eventually, the PCR amplicons were separated and detected by the homemade CE-ECL system. The detection of a typical genetically modified organism (GMO), Roundup Ready Soy (RRS), was shown as an example to demonstrate the reliability of the proposed approach. Four pairs of primers were amplified by multiple PCR (MPCR) simultaneously, three of which were targeted on the specific sequence of exogenous genes of RRS, and another was targeted on the endogenous reference gene of soybean. Both the conditions for PCR amplification and CE-ECL separation and detection were investigated in detail. Results showed that, under the optimal conditions, the proposed method can accurately identifying RRS. The corresponding limit of detection (LOD) was below 0.01% with 35 PCR cycles.

  17. Use of single cell gel electrophoresis assays for the detection of DNA-protective effects of dietary factors in humans: recent results and trends.

    Science.gov (United States)

    Hoelzl, Christine; Knasmüller, Siegfried; Misík, Miroslav; Collins, Andrew; Dusinská, Maria; Nersesyan, Armen

    2009-01-01

    This article summarises the results of human dietary intervention trials employing the comet assay (single cell gel electrophoresis, SCGE), which have been published in the last few years (i.e., between 2005 and 2008) and describes new trends and developments as well as current problems concerning the design of intervention trials and the interpretation of the results. Most new studies were carried out with complex plant derived foods and juices; only a few were conducted with individual food constituents. With specific vegetables, for example with water cress and Brussels sprouts, potent antioxidant effects were observed; also coffee caused a protective effect and it is notable that it was more effective than consumption of a diet containing increased levels of fruits and vegetables. Interesting recent developments include the development of protocols which enable us to monitor protection towards genotoxic chemicals contained in the human diet, and it was shown in preliminary studies that alterations of the activities of drug metabolising enzymes by dietary factors lead to altered sensitivity of lymphocytes against DNA damage caused by certain dietary carcinogens. Another novel approach is the development of methods to monitor the effects of dietary factors on DNA repair. The development of protocols for experiments with exfoliated buccal cells is another potentially valuable innovation. The adequate experimental design of SCGE trials is still a matter of debate and the evaluation of the available data shows that there is an urgent need to develop guidelines concerning the number of participants, sampling periods, duration of trials, use of placebos, and definition of adequate run-in and wash-out phases. Recent studies showed that the results of dietary studies could be biased by factors such as age, sex, body mass index and life style habits and by seasonal effects. Another still unsolved problem is the interpretation of the results of SCGE trials in regard to

  18. An investigation of the potential effect of sperm nuclear vacuoles in human spermatozoa on DNA fragmentation using a neutral and alkaline Comet assay.

    Science.gov (United States)

    Pastuszek, E; Kiewisz, J; Skowronska, P; Liss, J; Lukaszuk, M; Bruszczynska, A; Jakiel, G; Lukaszuk, K

    2017-03-01

    Presence of vacuoles and degree of sperm DNA damage are considered to be the basic factors used for the assessment of sperm fertilization capacity. We aimed to investigate the link between these two parameters. According to our knowledge, this is the first study where the Comet assay was used to assess the degree of DNA fragmentation of sperm categorized by Motile Sperm Organelle Morphology Examination (MSOME) Grades. Semen samples from 10 patients were assessed. Spermatozoa were graded into four MSOME groups according to the Vanderzwalmen's criteria. A total of 3930 motile spermatozoa were selected one-by-one using an inverted microscope and transferred onto two different slides. The degree of DNA fragmentation was analyzed by alkaline and neutral Comet assay. Results of the neutral Comet assay showed that Grade I spermatozoa (absence of vacuoles) presented significantly lower dsDNA fragmentation level (mean: 3.13 ± 1.17%) than Grade II (maximum of two small vacuoles; mean: 10.34 ± 2.65%), Grade III (more than two small vacuoles or at least one large vacuole; mean: 23.88 ± 8.37%), and Grade IV (large vacuoles associated with abnormal head shapes or other abnormalities; mean: 36.94 ± 7.78%; p  0.05). Probably, the vacuoles may be responsible for double strand DNA breaks rather than single strand DNA breaks (only 2.39% spermatozoa in MSOME Grade II, 1.76% in III, and 3.16% in IV has single strand breaks). The results demonstrate that lower MSOME grading correlates with lower sperm DNA fragmentation. Therefore, the observation of sperm nuclear vacuoles using real-time optical microscopy without precise DNA fragmentation examination is not sufficient for optimal sperm selection for intracytoplasmic sperm injection. © 2017 American Society of Andrology and European Academy of Andrology.

  19. Comet assay to determine genetic damage by the use of ivermectin in zebu cows (Bos taurus indicus

    Directory of Open Access Journals (Sweden)

    Donicer Montes-Vergara

    2017-05-01

    Full Text Available Objective. The objective of the work was evaluate the damage genetic caused by the use of ivermectin (IVM in cows zebu to concentrations of 1% and 3.15% through the test comet. Material and methods. 15 cows, were taken with age between 3 and 4 years old, average weight of 350 kg, body condition between 3 and 3.5. Three experimental groups with five animals per group, which were exposed to the concentration of IVM to 1% to 3.15% more group control (without application of IVM were used. Animal blood sample was performed by venipuncture jugular or medial flow with vacutainer® needle, extracting 8 ml of blood. The blood samples it was collected at 9, 18 and 27 days post-treatment. Results. The display of the comets is made by using fluorescence microscope, the cells were evaluated by means of visual log and the Comet image software. Evidenced the presence of nuclei with DNA migration in all analyzed plates. The values of classification of comets indicate cells with high levels of damage (grade 3: cells with high damage. The rate of DNA damage of the treatment to 1% to 3.15% was significant, to relate to the control group. Conclusions. The results obtained in this study demonstrate the likely genotoxic potential of the use of IVM in cattle.

  20. Chromatin structure influence of DNA damage measurements by four assays: pulsed- and constant-field gel electrophoresis, DNA precipitation and non-denaturing filter elution

    International Nuclear Information System (INIS)

    Wlodek, D.; Olive, P.L.

    1996-01-01

    The of elution of DNA during non-denaturing filter elution (NFE) often correlates with cell sensitivity to radiation. The elution rate is influenced by two cellular factors: chromatin structure and the number of DNA-strand breaks (DSBs) produced in an intact cell by ionizing radiation. To determine which of the above factors is relevant to cell radiosensitivity, four assays were used to measure induction of DNA damage in three cell lines varying in radiosensitivity (V79, CHO, and L5178Y-R). Each of the assays, neutral filter elution (NFE), DNA precipitation, constant (CFGE) and pulsed field gel electrophoresis (PFGE) have different physical basis for DNA damage measurement and might be differently affected by chromatin structure. Three of the methods used to measure DNA double-strand breaks gave different results: NFE was dependent on cell type and location of DNA relative to the replication fork, gel electrophoresis was independent of cell type but was affected by proximity to the replication fork, and the precipitation assay was independent of both cell type and replication status. Pulsed field gel electrophoresis produced the same results and constant field gel electrophoresis for 3 cell lines examined. Only NFE showed differences in sensitivity which correlated with cell survival following irradiation. The results suggest that three is the same initial amount of DSBs in cells from all three lines and that the sensitivity to radiation is determined by some additional factors, probably chromatin structure. (author). 18 refs, 5 figs

  1. First genotoxicity study of Paraná river water from Argentina using cells from the clam Corbicula fluminea (Veneroida Corbiculidae and Chinese hamster (Cricetulus griseus Rodentia, Cricetidae K1 cells in the comet assay

    Directory of Open Access Journals (Sweden)

    Jacqueline D. Caffetti

    2008-01-01

    Full Text Available High concentrations of xenobiotics from urban and industrial wastes have contributed to the contamination of many aquatic environments. We used the comet assay to evaluate the genotoxic potential of water collected from the River Paraná, which receives a great deal of waste, at three points (Puerto Piray, Eldorado and Montecarlo in the Misiones Province of Argentina. The in vivo comet assay used 40 freshwater clams (Corbicula fluminea while the in vitro comet assay used Chinese hamster (Cricetulus griseus K1 cell (CHO-K1 cultures with the mutagen ethyl methanesulfonate (EMS as the positive control and phosphate buffered saline (PBS as the negative control. Both assays showed statistically significant differences between the three sampling sites in relation to the negative control, the results of this preliminary study indicating that at these three sites water from the Paraná River presents genotoxic potential.

  2. Determination and assay validation of the bioactive sesquiterpene lactone xanthatin isolated from Xanthium cavanillesii using capillary electrophoresis.

    Science.gov (United States)

    Favier, L S; Acosta, G; Gomez, M R; María, A O; Tonn, C E

    2006-12-01

    This study describes the development and validation of a method for quantification of the antiulcer experimental drug xanthatin in tablets by capillary electrophoresis (CE). Solid oral dosage forms based on xanthatin were designed and assayed on rats. A CE methodology was developed; the parameters evaluated were: background electrolyte composition, concentration and pH, applied voltage and sample preparation. The method was validated in terms of range of linearity, limits of detection (LOD) and quantification (LOQ), accuracy, precision and selectivity and then applied to the pharmaceutical dosage forms. Xanthatin determination was carried out in less than 3 min with a 20 mM sodium tetraborate buffer, pH 9.20. Drug concentration per tablet found was 2.97 +/- 0.2 mg. Calibration plots were linear over at least three orders of magnitude of analyte concentrations, LOD and LOQ were 7.6 and 26 microg mL(-1) respectively. For accuracy evaluation a recovery test was performed, the values being better than 98.6%. With respect to precision, the results obtained were better than 1.02 RSD% (repeatability) and 1.54% (intermediate precision). After the manufacturing process the resulting tablets were biologically active. The methodology developed is useful, simple and rapid for xanthatin determination in tablets.

  3. Expression of Inflammatory and Cell Death Program Genes and Comet DNA Damage Assay Induced by Escherichia coli in Layer Hens

    Science.gov (United States)

    Mehaisen, Gamal M. K.; Eshak, Mariam G.; El Sabry, M. I.; Abass, Ahmed O.

    2016-01-01

    Modern methods of industrial poultry and egg production systems involve stressful practices that stimulate Escherichia coli (E. coli) activity causing endotoxic shock. This investigation was conducted to evaluate the expression of pro-inflammatory cytokines and cell death program genes and DNA damage induced by E. coli in the brain and liver tissues of laying hens. A total of two hundred and ten H&N brown layer hens with 20 week age, were used in this research. First, preliminary experiments were designed (60 hens in total) to establish the optimal exposure dose of E. coli and to determine the nearest time of notable response to be used in the remainder studies of this research. At 35-wk of age, 150 hens were randomly assigned into 2 groups with 3 replicates of 25 birds each; the first group was injected in the brachial wing vein with 107 E. coli colony/hen, while the second group was injected with saline and served as a control. The body temperature and plasma corticosterone concentration were measured 3 hr after injection. Specimens of liver and brain were obtained from each group and the gene expression of p38 mitogen-activated protein kinase, interlukin-1β (IL-1β), tumor necrosis factor alpha (TNF-α), Bax, and caspase-3 genes were measured by quantitative real-time PCR. DNA damage in the brain and liver tissues were also measured by comet assay. Hens treated with E. coli showed significant (P<0.05) increase of body temperature and plasma corticosterone (42.6°C and 14.5 ng/ml, respectively) compared to the control group (41.1°C and 5.5 ng/ml, respectively). Additional remarkable over-inflammation gene expression of p38, IL-1β and TNF-α.genes were also detected in the brain (2.2-fold, 2.0-fold and 3.3-fold, respectively) and the liver (2.1-fold, 1.9-fold and 3.0-fold, respectively) tissues of the infected chickens. It is also important to note that hens injected with E. coli showed an increase in DNA damage in the brain and liver cells (P<0.05). These

  4. Comet assay with gill cells of Mytilus galloprovincialis end point tools for biomonitoring of water antibiotic contamination: Biological treatment is a reliable process for detoxification.

    Science.gov (United States)

    Mustapha, Nadia; Zouiten, Amina; Dridi, Dorra; Tahrani, Leyla; Zouiten, Dorra; Mosrati, Ridha; Cherif, Ameur; Chekir-Ghedira, Leila; Mansour, Hedi Ben

    2016-04-01

    This article investigates the ability of Pseudomonas peli to treat industrial pharmaceuticals wastewater (PW). Liquid chromatography-mass spectrometry (MS)/MS analysis revealed the presence, in this PW, of a variety of antibiotics such as sulfathiazole, sulfamoxole, norfloxacine, cloxacilline, doxycycline, and cefquinome.P. peli was very effective to be grown in PW and inducts a remarkable increase in chemical oxygen demand and biochemical oxygen demand (140.31 and 148.51%, respectively). On the other hand, genotoxicity of the studied effluent, before and after 24 h of shaking incubation with P. peli, was evaluated in vivo in the Mediterranean wild mussels Mytilus galloprovincialis using comet assay for quantification of DNA fragmentation. Results show that PW exhibited a statistically significant (p< 0.001) genotoxic effect in a dose-dependent manner; indeed, the percentage of genotoxicity was 122.6 and 49.5% after exposure to 0.66 ml/kg body weight (b.w.); 0.33 ml/kg b.w. of PW, respectively. However, genotoxicity decreased strongly when tested with the PW obtained after incubation with P. peli We can conclude that using comet assay genotoxicity end points are useful tools to biomonitor the physicochemical and biological quality of water. Also, it could be concluded that P. peli can treat and detoxify the studied PW. © The Author(s) 2013.

  5. DNA damage induced by coal dust, fly and bottom ash from coal combustion evaluated using the micronucleus test and comet assay in vitro.

    Science.gov (United States)

    Matzenbacher, Cristina Araujo; Garcia, Ana Letícia Hilario; Dos Santos, Marcela Silva; Nicolau, Caroline Cardoso; Premoli, Suziane; Corrêa, Dione Silva; de Souza, Claudia Telles; Niekraszewicz, Liana; Dias, Johnny Ferraz; Delgado, Tânia Valéria; Kalkreuth, Wolfgang; Grivicich, Ivana; da Silva, Juliana

    2017-02-15

    Coal mining and combustion generating huge amounts of bottom and fly ash are major causes of environmental pollution and health hazards due to the release of polycyclic aromatic hydrocarbons (PAH) and heavy metals. The Candiota coalfield in Rio Grande do Sul, is one of the largest open-cast coal mines in Brazil. The aim of this study was to evaluate genotoxic and mutagenic effects of coal, bottom ash and fly ash samples from Candiota with the comet assay (alkaline and modified version) and micronucleus test using the lung fibroblast cell line (V79). Qualitative and quantitative analysis of PAH and inorganic elements was carried out by High Performance Liquid Chromatography (HPLC) and by Particle-Induced X-ray Emission (PIXE) techniques respectively. The samples demonstrated genotoxic and mutagenic effects. The comet assay modified using DNA-glicosilase formamidopirimidina (FPG) endonuclease showed damage related to oxidative stress mechanisms. The amount of PAHs was higher in fly ash followed by pulverized coal. The amount of inorganic elements was highest in fly ash, followed by bottom ash. It is concluded that the samples induce DNA damage by mechanisms that include oxidative stress, due to their complex composition, and that protective measures have to be taken regarding occupational and environmental hazards. Copyright © 2016 Elsevier B.V. All rights reserved.

  6. Genetic damage induced by trophic doses of lead in the neotropical fish Hoplias malabaricus (Characiformes, Erythrinidae as revealed by the comet assay and chromosomal aberrations

    Directory of Open Access Journals (Sweden)

    Marta Margarete Cestari

    2004-01-01

    Full Text Available The effects of clastogenic or mutagenic agents have rarely been studied in neotropical fish species exposed to contaminated water. In this study, the genetic damage caused by lead in the widely distributed South American fish, Hoplias malabaricus, was assessed using the comet (SCGE assay and by testing for chromosomal aberrations. Eighteen specimens were acclimatized to laboratory conditions and then chronically exposed to contaminated food by feeding prey (Cyprinus sp. injected intraperitoneally with doses of inorganic lead adjusted to give a contamination level of 21 mg of Pb2+.g-1 net weight of H. malabaricus. Three fish were sampled for chromosomal analysis after four doses (18 days and another three after eight doses (41 days of lead and the results then compared with three untreated controls kept under lead-free conditions. An additional six treated fish and three controls were sampled for the comet assay after 13 doses (64 days. Exposure to lead significantly increased the frequency of chromosomal aberrations and the frequency of tailed cell nuclei, the latter indicating DNA damage. These results show that H. malabaricus is a useful biological model for screening the clastogenic effects of lead and possibly other xenobiotics. The genetic damage seen here illustrates the need to investigate the potential effects of heavy metals on fish species in South America.

  7. Detection of genotoxicity of water from an urbanized stream, in Corbicula fluminea (Mollusca) (in vivo) and CHO-K1 cells (in vitro) using comet assay.

    Science.gov (United States)

    Rigonato, Janaina; Mantovani, Mário Sérgio; Jordão, Berenice Quinzani

    2010-07-01

    The comet assay was utilized to investigate the quality of water from seven locations along the Cambé Stream, in vivo (Corbicula fluminea hemolymph), in vitro (CHO-K1 cells), in situ, and in laboratory studies. The Cambé Stream basin (Londrina, PR, Brazil) is almost completely urbanized and receives different forms of industrial and domestic runoff. The data indicated the occurrence of DNA damage in cells examined in vivo and in vitro, shown by the significant increase in frequencies of cells with DNA damage after exposure to water from all seven locations used in the study. Our results strongly suggest the presence of genotoxic agent(s) in water at all of the sampled locations, demonstrated by elevated numbers of cells with DNA damaged in field and laboratory tests. In all of the places sampled, domestic sewage influence appeared to be one important cause for the introduction of xenobiotics, environmental genotoxins, and pollutants into the water. Thus, the comet assay applied in these cell systems was able to detect adverse environmental conditions, proving to be a very adequate short-term test and should be included in batteries of tests utilized in the monitoring of aquatic environments.

  8. Free radical-scavenging activity and DNA damaging potential of auxins IAA and 2-methyl-IAA evaluated in human neutrophils by the alkaline comet assay.

    Science.gov (United States)

    Salopek-Sondi, Branka; Piljac-Zegarac, Jasenka; Magnus, Volker; Kopjar, Nevenka

    2010-01-01

    Auxins, of which indole-3-acetic acid (IAA) is the most widespread representative, are plant hormones. In addition to plants, IAA also naturally occurs in humans in micromolar concentrations. In the presence of peroxidase, indolic auxins are converted to cytotoxic oxidation products and have thus been proposed for use in gene-directed enzyme/prodrug tumor therapy. Since data on the genotoxicity of IAA and its derivatives are not consistent, here we investigate the early DNA damaging effects (2-h treatment) of the auxins, IAA, and 2-methyl-indole-3-acetic acid (2-Me-IAA) by the alkaline comet assay and compare them with their free radical-scavenging activity measured by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. Human neutrophils are chosen as the test system since they possess inherent peroxidase activity. The results of the comet assay indicate an increase in DNA damage in a dose-dependent manner up to 1.00 mM of both auxins. Generally, IAA applied in the same concentration had greater potential to damage DNA in human neutrophils than did 2-Me-IAA. The genotoxicities of the two examined auxins are negatively correlated with their antioxidant activities, as measured by the DPPH assay; 2-Me-IAA showed a higher antioxidant capacity than did IAA. We assume that differences in the molecular structure of the tested auxins contributed to differences in their metabolism, in particular, with respect to interactions with peroxidases and other oxidative enzymes in neutrophils. However, the exact mechanisms have to be elucidated in future studies. (c) 2010 Wiley Periodicals, Inc.

  9. DNA damage induced by indirect and direct acting mutagens in catalase-deficient transgenic tobacco Cellular and acellular Comet assays

    Czech Academy of Sciences Publication Activity Database

    Gichner, Tomáš

    2003-01-01

    Roč. 535, - (2003), s. 187-193 ISSN 1383-5718 R&D Projects: GA ČR GA521/02/0400 Institutional research plan: CEZ:AV0Z5038910 Keywords : Hydrogen peroxide * Single-cell gel electrophoresis * Nicotiana tabacum Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 1.748, year: 2003

  10. Cytokinesis-blocked micronucleus cytome and comet assays in peripheral blood lymphocytes of workers exposed to lead considering folate and vitamin B12 status.

    Science.gov (United States)

    Minozzo, Renato; Deimling, Luiz Irineu; Santos-Mello, Renato

    2010-03-29

    In human biomonitoring, factors such as age, gender, smoking habit and alcohol consumption are usually considered and identified as agents that exert an impact on genotoxicity biomarkers. However, as a rule, factors like micronutrient status are not considered in biomonitoring studies. This paper reports on genotoxic damage in Pb-exposed workers using data obtained from the cytokinesis-block micronucleus (CBMN) cytome assay and alkaline comet assay with silver staining, considering folate and vitamin B12 nutritional status. Analysis of the results showed that the Pb-exposed group presented a 24-fold higher Pb content in the blood compared to controls. The Pb-exposed workers presented significantly greater micronuclei (MNi) counts (Z=7.9583; p<0.0001) and DNA damage, assessed by the comet assay (DF: Z=7.7056; p<0.0001 and DI: Z=7.4749; p<0.0001), but no correlation with Pb blood concentrations were detected. These two groups did not differ significantly concerning folate and vitamin B12 levels. Regarding folate, it is possible that this similarity is associated with folic acid flour-enrichment, introduced in Brazil in 2004 to prevent neural tube defects. The mean folate level obtained in the Pb-exposed group was 6.18ng/ml. The subgroup of individuals with serum folate levels equal to or higher than the mean presented significantly higher MNi (Z=2.3776; p=0.017) and nucleoplasmic bridge (NPB) frequencies (Z=1.9850; p=0.047) in peripheral blood lymphocytes. A significant positive correlation was observed between the age of Pb-exposed workers and MNi frequencies (Gamma=0.3328; p<0.001), NPBs (Gamma=0.1832; p<0.042) and DNA damage assessed by the comet assay (DF: Gamma=0.1764; p=0.035 and DI: Gamma=0.11852; p=0.028). These findings suggest that high folate levels alone do not guarantee protection against genotoxic damage. Moreover, folic acid supplementation should be studied using more efficient approaches to determine safe amounts and potential deleterious effects

  11. Observing comets

    CERN Document Server

    James, Nick

    2003-01-01

    Since comet Shoemaker-Levy collided with the planet Jupiter with stupendous force in 1994 there has been an upsurge of amateur interest in comets Most comets are first discovered by amateur astronomers because there are so many amateurs looking for them, and techniques and instruments have improved dramatically in the past few years After a short but detailed introduction to the comets themselves Nick James and Gerald North describe comet hunting, photographing and imaging comets, and digital image processing The use of computers for orbital calculations and even helping to discover new comets is given a full chapter, as are advanced techniques including comet photometry and spectroscopy This comprehensive book has an accompanying CD-ROM and is at once a "primer" for comet hunters and a reference text for more advanced amateur astronomers

  12. Use of the Comet test and micronucleus assay on human white blood cells for in vitro assessment of genotoxicity induced by different drinking water disinfection protocols.

    Science.gov (United States)

    Maffei, Francesca; Buschini, Annamaria; Rossi, Carlo; Poli, Paola; Forti, Giorgio Cantelli; Hrelia, Patrizia

    2005-08-01

    Surface water disinfection can lead to the formation of mutagenic/carcinogenic by-products derived from reactions with naturally occurring inorganic compounds. We investigated the feasibility and potential usefulness of an integrated approach to genotoxicity analysis of drinking water. The approach employed the Comet and micronucleus (MN) assays to evaluate the DNA and chromosomal damage produced by water extracts in human blood cells. Surface water samples from Lago Trasimeno (Italy) were collected in different seasons (July 2000, October 2000, February 2001, and June 2001), and samples were disinfected with sodium hypochloride (NaClO), chlorine dioxide (ClO(2)), or peracetic acid (PAA). Extracts of untreated and treated water were incubated with primary human leukocytes. The Comet assay revealed both strong seasonal variations and differences between samples processed by the three disinfection protocols. The three disinfectants increased the genotoxicity of the water collected in July 2000 and October 2000, with PAA producing the greatest amount of DNA damage. Extracts of raw water collected in February 2001 produced so much DNA damage that the relative genotoxic potentials of the three disinfectants could not be evaluated. No increase in MN frequency was detected in any of the samples. The multi-endpoint MN assay indicated, however, that our study samples (especially the sample collected in the February 2001) were cytotoxic. We conclude that this integrated approach to genotoxicity assessment may be useful both for the quality control of raw drinking water and to help compare the potential health risks associated with alternative disinfection processes. Copyright 2005 Wiley-Liss, Inc.

  13. An alkaline comet assay study on the antimalarial drug atovaquone in human peripheral blood lymphocytes: a study based on clinically relevant concentrations.

    Science.gov (United States)

    Dinter, Domagoj; Gajski, Goran; Garaj-Vrhovac, Vera

    2013-01-01

    Atovaquone, a hydroxynaphthoquinone, is an anti-parasite drug, selectively targeting the mitochondrial respiratory chain of malaria parasite. It is used for both the treatment and prevention of malaria, usually in a fixed combination with proguanil. Although atovaquone has not often been associated with severe adverse reactions in the recommended dosages and has a relatively favorable side effect profile, the present study was undertaken to evaluate its cytogenotoxic potential towards human peripheral blood lymphocytes. Two different concentrations of atovaquone found in plasma when used in fixed-dose combination with proguanile hydrochloride were used with and without S9 metabolic activation: 2950 ng ml(-1) used for prophylactic treatment and 11 800 ng ml(-1) used in treatment of malaria. The results showed that lymphocyte viability was not affected after the treatment, suggesting that atovaquone was not cytotoxic in the given concentrations. With the alkaline comet assay we demonstrated that in human peripheral blood lymphocytes no significant changes in comet parameters occurred after the treatment. There were no differences in tested parameters with the addition of S9 metabolic activation, indicating that atovaquone either has no metabolite or it is not toxic in the given concentrations. Since no effects were observed after the treatment, it is to be concluded that atovaquone is safe from the aspect of genototoxicity in the recommended dosages. Copyright © 2011 John Wiley & Sons, Ltd.

  14. Differential genotoxicity of ethyl methanesulphonate, N-ethyl-N-nitrosourea and maleic hydrazide in tobacco seedlings based on data of the Comet assay and two recombination assays

    Czech Academy of Sciences Publication Activity Database

    Gichner, Tomáš

    2003-01-01

    Roč. 538, - (2003), s. 171-179 ISSN 1383-5718 R&D Projects: GA ČR GA521/02/0400; GA MŠk LN00B030 Institutional research plan: CEZ:AV0Z5038910 Keywords : DNA damage * Nicotiana tabacum * Single cell gel electrophoresis (SCGE) Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 1.748, year: 2003

  15. Multiplex polymerase chain reaction-capillary gel electrophoresis: a promising tool for GMO screening--assay for simultaneous detection of five genetically modified cotton events and species.

    Science.gov (United States)

    Nadal, Anna; Esteve, Teresa; Pla, Maria

    2009-01-01

    A multiplex polymerase chain reaction assay coupled to capillary gel electrophoresis for amplicon identification by size and color (multiplex PCR-CGE-SC) was developed for simultaneous detection of cotton species and 5 events of genetically modified (GM) cotton. Validated real-time-PCR reactions targeting Bollgard, Bollgard II, Roundup Ready, 3006-210-23, and 281-24-236 junction sequences, and the cotton reference gene acp1 were adapted to detect more than half of the European Union-approved individual or stacked GM cotton events in one reaction. The assay was fully specific (labeling in one step. Thus, it is an easy and inexpensive tool for initial screening, to be complemented with quantitative assays if necessary.

  16. Desenvolvimento e validação de técnica quantitativa de análise de imagem para avaliação do teste do cometa corado pela prata Development and validation of a quantitative image analysis method to evaluate comet assay (silver staining

    Directory of Open Access Journals (Sweden)

    Gabrielli Brianezi

    2009-08-01

    Full Text Available INTRODUÇÃO: O ensaio do cometa ou técnica da eletroforese de células isoladas é largamente empregado para avaliação de danos e reparo do DNA em células individuais. O material pode ser corado por técnicas de fluorescência ou por sal de prata. Este último apresenta vantagens técnicas, como o tipo de microscópio utilizado e a possibilidade de armazenamento das lâminas. A análise dos cometas pode ser feita de modo visual, porém há a desvantagem da subjetividade dos resultados, que pode ser minimizada por análise digital automatizada. OBJETIVOS: Desenvolvimento e validação de método de análise digital de cometas corados por sal de prata. MÉTODOS: Cinquenta cometas foram fotografados de maneira padronizada e impressos em papel. Além de medidas manualmente, essas imagens foram classificadas em cinco categorias por três avaliadores, antes e depois de pré-processadas automaticamente pelo software ImageJ 1.38x. As estimativas geradas pelos avaliadores foram comparadas quanto sua correlação e reprodutibilidade. Em seguida, foram desenvolvidos algoritmos de análise digital das medidas, com base em filtros estatísticos de mediana e de mínimo. Os valores obtidos foram comparados com os estimados manual e visualmente após o pré-processamento. RESULTADOS: As medidas manuais das imagens pré-processadas apresentaram maior correlação intraclasse do que as imagens preliminares. Os parâmetros automatizados apresentaram alta correlação com as medidas manuais pré-processadas, sugerindo que este sistema aumenta a objetividade da análise, podendo ser utilizado na estimativa dos parâmetros dos cometas. CONCLUSÃO: A presente análise digital proposta para o teste do cometa corado pela prata mostrou-se factível e de melhor reprodutibilidade que a análise visual.BACKGROUND: Comet assay or single cell gel electrophoresis is a useful and widely applied technique for the assessment of DNA damage and repair in individual cells

  17. A neutral glyoxal gel electrophoresis method for the detection and semi-quantitation of DNA single-strand breaks.

    Science.gov (United States)

    Pachkowski, Brian; Nakamura, Jun

    2013-01-01

    Single-strand breaks are among the most prevalent lesions found in DNA. Traditional electrophoretic methods (e.g., the Comet assay) used for investigating these lesions rely on alkaline conditions to denature DNA prior to electrophoresis. However, the presence of alkali-labile sites in DNA can result in the introduction of additional single-strand breaks upon alkali treatment during DNA sample processing. Herein, we describe a neutral glyoxal gel electrophoresis assay which is based on alkali-free DNA denaturation and is suitable for qualitative and semi-quantitative analyses of single-strand breaks in DNA isolated from different organisms.

  18. Great Comets

    Science.gov (United States)

    Burnham, Robert

    2000-05-01

    Spectacular and mysterious objects that come and go in the night sky, comets have dwelt in our popular culture for untold ages. As remnants from the formation of the Solar system, they are objects of key scientific research and space missions. As one of nature's most potent and dramatic dangers, they pose a threat to our safety--and yet they were the origin of our oceans and perhaps even life itself. This beautifully illustrated book tells the story of the biggest and most awe-inspiring of all comets: those that have earned the title "Great." Robert Burnham focuses on the Great comets Hyakutake in 1996 and Hale-Bopp in 1997, which gripped attention worldwide because, for many, they were the first comets ever seen. He places these two recent comets in the context of their predecessors from past ages, among them the famous Comet Halley. Great Comets explains the exciting new discoveries that have come from these magnificent objects and profiles the spaceprobes to comets due for launch in the next few years. The book even takes a peek behind Hollywood's science-fiction fantasies to assess the real risks humanity faces from potential impacts of both comets and asteroids. For everyone interested in astronomy, this exciting book reveals the secrets of the Great Comets and provides essential tools for keeping up to date with comet discoveries in the future. Robert Burnham has been an amateur astronomer since the mid-1950s. He has been a senior editor of Astronomy magazine (1986-88) and is the author of many books and CD-ROMS, including Comet Hale-Bopp: Find and Enjoy the Great Comet and Comet Explorer.

  19. Human MLPA Probe Design (H-MAPD: a probe design tool for both electrophoresis-based and bead-coupled human multiplex ligation-dependent probe amplification assays

    Directory of Open Access Journals (Sweden)

    Hatchwell Eli

    2008-09-01

    Full Text Available Abstract Background Multiplex ligation-dependent probe amplification (MLPA is an efficient and reliable technique for gene dosage analysis. Currently MLPA can be conducted on two platforms: traditional electrophoresis-based, and FlexMAP bead-coupled. Since its introduction in 2002, MLPA has been rapidly adopted in both clinical and research situations. However, MLPA probe design is a time consuming process requiring many steps that address multiple criteria. There exist only one or two commercial software packages for traditional electrophoresis-based MLPA probe design. To our knowledge, no software is yet available that performs bead-coupled MLPA probe design. Results We have developed H-MAPD, a web-based tool that automates the generation and selection of probes for human genomic MLPA. The software performs physical-chemical property tests using UNAFold software, and uniqueness tests using the UCSC genome browser. H-MAPD supports both traditional electrophoresis-based assays, as well as FlexMAP bead-coupled MLPA. Conclusion H-MAPD greatly reduces the efforts for human genomic MLPA probe design. The software is written in Perl-CGI, hosted on a Linux server, and is freely available to non-commercial users.

  20. Genotoxicity of a thiosulfonate compound derived from Allium sp. intended to be used in active food packaging: In vivo comet assay and micronucleus test.

    Science.gov (United States)

    Mellado-García, Pilar; Puerto, María; Prieto, Ana I; Pichardo, Silvia; Martín-Cameán, Ana; Moyano, Rosario; Blanco, Alfonso; Cameán, Ana M

    2016-04-01

    Components of Allium species have antimicrobial and antioxidant properties. A commercial Allium sp. extract (Proallium AP(®)), of which the main constituent is propyl thiosulphinate oxide (PTSO), is being used in the development of active food packaging. In previous in vitro genotoxicity studies, PTSO, in the presence of metabolic activation, increased the appearance of micronuclei (MN). We assessed the genotoxicity PTSO in rats following oral administration (doses: 5.5, 17.4, and 55mg/kg). The comet assay in liver and stomach (OECD 489) and the MN assay in bone marrow (OECD 474) were carried out. After necropsy, histopathological examinations of the liver and the stomach were performed. The results revealed no in vivo genotoxicity and the histopathological analysis showed only slight modifications, such as increased glycogen storage in the liver and a degenerative process in stomach, with vacuolization of cell membranes, only at the highest dose. Therefore, the present work confirms that this compound is not genotoxic and could be considered as a natural alternative to synthetic preservatives used in the food packaging industry. Copyright © 2016 Elsevier B.V. All rights reserved.

  1. Assay of phenolic compounds in red wine by on-line combination of capillary isotachophoresis with capillary zone electrophoresis

    Czech Academy of Sciences Publication Activity Database

    Hamoudová, R.; Urbánek, Marek; Pospíšilová, M.; Polášek, M.

    2004-01-01

    Roč. 1032, 1-2 (2004), s. 281-287 ISSN 0021-9673 R&D Projects: GA MZd NL7689 Grant - others:GA FR VŠ(CZ) G6/2995/2003 Institutional research plan: CEZ:AV0Z4031919 Keywords : wine * food analysis * isotachophoresis * capillary zone electrophoresis Subject RIV: CB - Analytical Chemistry, Separation Impact factor: 3.359, year: 2004

  2. Detection of Phakopsora pachyrhizi by polymerase chain reaction (PCR) and use of germination test and DNA comet assay after e-beam processing in soybean

    International Nuclear Information System (INIS)

    Villavicencio, A.L.C.H.; Fanaro, G.B.; Araujo, M.M.; Aquino, S.; Silva, P.V.; Mancini-Filho, J.

    2007-01-01

    Soybean harvest is the main agribusiness in Brazil, which is the second largest exporter in the world and has a revenue of billions of dollars. Asian dust is caused by the fungus Phakopsora pachyrhizi and its dissemination is difficult to control, since it occurs through wind dispersion. Actually P. pachyrhizi is found in different parts of the world. Electron beam treatment could be an alternative process to minimize these losses, especially for the grains exportation industry. Besides the possibility of being disconnected when not in use, this source does not need to be reloaded, is easily available and, streamlines the process and reduces logistics costs. The present work aims to identify, by the polymerase chain reaction technique (PCR), the P. pachyrhizi fungus presence in the irradiated soybeans and the possibility to use radiation treatment as a sanitary alternative. Doses 0, 1.0, 2.0, 5.0, 6.0, 7.0, 8.0, 9.0 and 10.0 kGy (IPEN-CNEN/SP Electron Accelerator) were applied and two fast-screening methods were performed: DNA comet assay (for the detection of DNA damage) and germination test (for the measurement of roots inhibition). These tests are very easy to carry out and measure damage response depending on radiation dose

  3. In vivo genotoxicity evaluation of atrazine and atrazine-based herbicide on fish Carassius auratus using the micronucleus test and the comet assay.

    Science.gov (United States)

    Cavas, Tolga

    2011-06-01

    Atrazine is a selective triazine herbicide used to control broadleaf and grassy weeds mainly in corn, sorghum, sugarcane, pineapple, and other crops, and in conifer reforestation planting fields. It has been showed that atrazine is one of the most frequently detected pesticides in agricultural streams and rivers, over the past two decades. Although the toxic properties of atrazine are well known, the data on the genotoxic effects of atrazine on aquatic organisms are rather scarce. Thus, in the present study we aimed to evaluate the genotoxic effects of atrazine and an atrazine-based herbicide (Gesaprim®) on a model fish species Carassius auratus L., 1758, (Pisces: Cyprinidae) using the micronucleus test and the comet assay in peripheral blood erythrocytes. Fish were exposed to 5, 10 and 15 μg/L atrazine and to its commercial formulation for 2, 4 and 6 days. Ethyl methane sulfonate (EMS) at a single dose of 5 mg/L was used as positive control. Our results revealed significant increases in the frequencies of micronuclei and DNA strand breaks in erythrocytes of C. auratus, following exposure to commercial formulation of atrazine and thus demonstrated the genotoxic potential of this pesticide on fish. Copyright © 2011 Elsevier Ltd. All rights reserved.

  4. DNA damage in isolated rat hepatocytes exposed to C.I. pigment orange 5 and C.I. pigment yellow 12 by the alkaline comet assay

    DEFF Research Database (Denmark)

    Møller, P; Wallin, Håkan; Grunnet, N

    1998-01-01

    , and C.I. pigment orange 5 is a naphthol-azo pigment. The pigments are virtually insoluble in aqueous solutions, and they have not been tested extensively for toxicological effects. C.I. pigment orange 5 increased the levels of DNA damage at 5 microg/ml (P pigment yellow 12 at 20 microg....../ml (P effect of incubation time (20, 40, and 80 min) of the same concentrations of the pigments was tested. The levels of DNA damage were increased up to 80 min. Both pigments produced DNA damage that was in the same range as the food carcinogen 2-amino-3,8-dimethylimidazo[4,5-f......The induction of DNA damage by commonly used printing ink pigments, C.I. pigment orange 5 (C.I. 12075) and C.I. pigment yellow 12 (C.I. 21090), was investigated in freshly isolated rat hepatocytes with the comet assay. C.I. pigment yellow 12 is a 3,3'-dichlorobenzidine-based diarylide pigment...

  5. Zinc-induced DNA damage and the distribution of metals in the brain of grasshoppers by the comet assay and micro-PIXE.

    Science.gov (United States)

    Augustyniak, Maria; Juchimiuk, Jolanta; Przybyłowicz, Wojciech J; Mesjasz-Przybyłowicz, Jolanta; Babczyńska, Agnieszka; Migula, Paweł

    2006-11-01

    The distribution and concentration of selected elements by PIXE method and DNA damage using comet assay in brains of 1st instars of grasshoppers Chorthippus brunneus from unpolluted (Pilica) and polluted (Olkusz) site, additionally exposed to various doses of zinc during diapause or after hatching, were measured. We tried to assess the degree of possible pre-adaptation of the insects to heavy metals and evaluate the utility of these parameters in estimation of insect exposure to industrial pollutants. Additionally, the mechanism of zinc toxicity for grasshopper brains was discussed. We observed the correlation between experimental zinc dose, zinc contents in the brain and DNA damage in neuroblasts, but only in groups exposed to lower zinc concentration. For higher zinc concentration the amount of the metal in brain and DNA damage remained at the control level. Some site-related differences in DNA damage between grasshoppers from Pilica and Olkusz were observed during short-term exposure (after hatching). Significant increase in the calcium contents in the brain, proportional to zinc concentration in sand, was also observed, especially in the offsprings from Olkusz. The results may be the basis for further searching for molecular mechanisms of defense against heavy metals in insects living in polluted habitats.

  6. Studying the synergistic damage effects induced by 1.8 GHz radiofrequency field radiation (RFR) with four chemical mutagens on human lymphocyte DNA using comet assay in vitro

    International Nuclear Information System (INIS)

    Wang Baohong; He Jiliang; Jin Lifen; Lu Deqiang; Zheng Wei; Lou Jianlin; Deng Hongping

    2005-01-01

    The aim of this investigation was to study the synergistic DNA damage effects in human lymphocytes induced by 1.8 GHz radiofrequency field radiation (RFR, SAR of 3 W/kg) with four chemical mutagens, i.e. mitomycin C (MMC, DNA crosslinker), bleomycin (BLM, radiomimetic agent), methyl methanesulfonate (MMS, alkylating agent), and 4-nitroquinoline-1-oxide (4NQO, UV-mimetic agent). The DNA damage of lymphocytes exposed to RFR and/or with chemical mutagens was detected at two incubation time (0 or 21 h) after treatment with comet assay in vitro. Three combinative exposure ways were used. Cells were exposed to RFR and chemical mutagens for 2 and 3 h, respectively. Tail length (TL) and tail moment (TM) were utilized as DNA damage indexes. The results showed no difference of DNA damage indexes between RFR group and control group at 0 and 21 h incubation after exposure (P > 0.05). There were significant difference of DNA damage indexes between MMC group and RFR + MMC co-exposure group at 0 and 21 h incubation after treatment (P 0.05). The experimental results indicated 1.8 GHz RFR (SAR, 3 W/kg) for 2 h did not induce the human lymphocyte DNA damage effects in vitro, but could enhance the human lymphocyte DNA damage effects induced by MMC and 4NQO. The synergistic DNA damage effects of 1.8 GHz RFR with BLM or MMS were not obvious

  7. Protective effect of dry olive leaf extract in adrenaline induced DNA damage evaluated using in vitro comet assay with human peripheral leukocytes.

    Science.gov (United States)

    Cabarkapa, Andrea; Zivković, Lada; Zukovec, Dijana; Djelić, Ninoslav; Bajić, Vladan; Dekanski, Dragana; Spremo-Potparević, Biljana

    2014-04-01

    Excessive release of stress hormone adrenaline is accompanied by generation of reactive oxygen species which may cause disruption of DNA integrity leading to cancer and age-related disorders. Phenolic-rich plant product dry olive leaf extract (DOLE) is known to modulate effects of various oxidants in human cells. The aim was to evaluate the effect of commercial DOLE against adrenaline induced DNA damage in human leukocytes by using comet assay. Peripheral blood leukocytes from 6 healthy subjects were treated in vitro with three final concentrations of DOLE (0.125, 0.5, and 1mg/mL) for 30 min at 37°C under two different protocols, pretreatment and post-treatment. Protective effect of DOLE was assessed from its ability to attenuate formation of DNA lesions induced by adrenaline. Compared to cells exposed only to adrenaline, DOLE displayed significant reduction (Padrenaline genotoxicity. Results indicate genoprotective and antioxidant properties in dry olive leaf extract, strongly supporting further explorations of its underlying mechanisms of action. Copyright © 2014 Elsevier Ltd. All rights reserved.

  8. Detection of Phakopsora pachyrhizi by polymerase chain reaction (PCR) and use of germination test and DNA comet assay after e-beam processing in soybean

    Energy Technology Data Exchange (ETDEWEB)

    Villavicencio, A.L.C.H. [Instituto de Pesquisas Energeticas e Nucleares-IPEN-CNEN/SP, Centro de Tecnologia das Radiacoes, Lab. de Deteccao de Alimentos Irradiados, Cidade Universitaria, Av. Prof. Lineu Prestes 2242, Butanta CEP 05508-910, Sao Paulo (Brazil)], E-mail: villavic@ipen.br; Fanaro, G.B.; Araujo, M.M.; Aquino, S.; Silva, P.V. [Instituto de Pesquisas Energeticas e Nucleares-IPEN-CNEN/SP, Centro de Tecnologia das Radiacoes, Lab. de Deteccao de Alimentos Irradiados, Cidade Universitaria, Av. Prof. Lineu Prestes 2242, Butanta CEP 05508-910, Sao Paulo (Brazil); Mancini-Filho, J. [Faculdade de Ciencias Farmaceuticas-FCF/USP, Departamento de Alimentos e Nutricao Experimental, Lab. de Lipides, Sao Paulo (Brazil)

    2007-11-15

    Soybean harvest is the main agribusiness in Brazil, which is the second largest exporter in the world and has a revenue of billions of dollars. Asian dust is caused by the fungus Phakopsora pachyrhizi and its dissemination is difficult to control, since it occurs through wind dispersion. Actually P. pachyrhizi is found in different parts of the world. Electron beam treatment could be an alternative process to minimize these losses, especially for the grains exportation industry. Besides the possibility of being disconnected when not in use, this source does not need to be reloaded, is easily available and, streamlines the process and reduces logistics costs. The present work aims to identify, by the polymerase chain reaction technique (PCR), the P. pachyrhizi fungus presence in the irradiated soybeans and the possibility to use radiation treatment as a sanitary alternative. Doses 0, 1.0, 2.0, 5.0, 6.0, 7.0, 8.0, 9.0 and 10.0 kGy (IPEN-CNEN/SP Electron Accelerator) were applied and two fast-screening methods were performed: DNA comet assay (for the detection of DNA damage) and germination test (for the measurement of roots inhibition). These tests are very easy to carry out and measure damage response depending on radiation dose.

  9. 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.

  10. The DNA comet assay and the germination test in detection of food treated by ionizing radiation; Teste do cometa e teste de germinacao na deteccao do tratamento de alimentos com a radiacao ionizante

    Energy Technology Data Exchange (ETDEWEB)

    Huachaca, Nelida Simona Marin

    2002-07-01

    Two methods of irradiated food detection, one biochemical, the comet assay and, other biological, the germination test, were applied in bovine meat and fruit samples. The comet assay detects the damage on DNA caused by ionizing radiation. The germination test evaluates the sensitivity to radiation of seeds as for germination ability, shooting and, rooting. The samples were irradiated in gamma font and electron accelerator. For bovine meat samples, the doses were 0.0; 2.5; 4.5 e 7.0 kGy at chilled condition and, 0.0; 2.5; 4.5; 7.0 e 8.5 kGy at frozen conditions. For fruit samples such as melon, watermelon, apple, orange, papaya and, tomato, the doses were: 0.0; 0.5; 0.75; 1.0; 2.0 e 4.0 kGy. The differences between the gamma rays and the electron beam effects on extent of DNA migration and, on shooting and rooting, showed to be similar. The comet assay, under neutral conditions, permitted to discriminate between irradiated and unirradiated bovine meat samples, until one month of storage. Also, it was possible to distinguish, by the comet assay, the control sample with regard to irradiated fruit, at doses as low as 0,5 kGy. In the germination test, the root length was the best parameter to discriminate irradiated and unirradiated samples of melon, watermelon and tomato, while the germination percent was the best parameter for apple and orange. (author)

  11. Toxicity evaluation of water samples collected near a hospital waste landfill through bioassays of genotoxicity piscine micronucleus test and comet assay in fish Astyanax and ecotoxicity Vibrio fischeri and Daphnia magna.

    Science.gov (United States)

    Erbe, Margarete Casagrande Lass; Ramsdorf, Wanessa Algarte; Vicari, Taynah; Cestari, Marta Margarete

    2011-03-01

    In this study, we analyzed samples of water from a river and a lake located near a hospital waste landfill with respect to physico-chemical parameters and conducted bioassays of ecotoxicity using Vibrio fischeri and Daphnia magna, which are species commonly used to evaluate the water toxicity. We also evaluated damage to the genetic material of fish (Astyanax sp. B) that were exposed (96 h) to water from these two sites that were located near the tank ditch, using the alkaline comet assay and the piscine micronucleus test. Parameters including aluminum, manganese, biochemical oxygen demand, sulfide, conductivity, phenol, total coliforms and Escherichia coli counts, were above acceptable levels that have been established in environmental legislation. However, the toxicity bioassays that we carried out in Vibrio fischeri and Daphnia magna and the piscine micronucleus test in fish showed no immediate risk due to acute effects. Based on the results of the comet assay, however, it was possible to detect damage to genetic material in fish that were acutely exposed in the laboratory to water samples from the river and lake that are located near the trench septic tank. Thus, our results suggest that tests beyond those usually employed to test water toxicity, such as the comet assay we used in the fish, are required to assess the toxicity of water with greater accuracy.

  12. Effects of motexafin gadolinium on DNA damage and X-ray-induced DNA damage repair, as assessed by the Comet assay

    International Nuclear Information System (INIS)

    Donnelly, Erling T.; Liu Yanfeng; Paul, Tracy K.; Rockwell, Sara

    2005-01-01

    Purpose: To investigate the effects of motexafin gadolinium (MGd) on the levels of reactive oxygen species (ROS), glutathione (GSH), and DNA damage in EMT6 mouse mammary carcinoma cells. The ability of MGd to alter radiosensitivity and to inhibit DNA damage repair after X-ray irradiation was also evaluated. Methods and Materials: Reactive oxygen species and GSH levels were assessed by 2,7-dichlorofluorescein fluorescence flow cytometry and the Tietze method, respectively. Cellular radiosensitivity was assessed by clonogenic assays. Deoxyribonucleic acid damage and DNA damage repair were assessed in plateau-phase EMT6 cells by the Comet assay and clonogenic assays. Results: Cells treated with 100 μmol/L MGd plus equimolar ascorbic acid (AA) had significantly increased levels of ROS and a 58.9% ± 3.4% decrease in GSH levels, relative to controls. Motexafin gadolinium plus AA treatment increased the hypoxic, but not the aerobic, radiosensitivity of EMT6 cells. There were increased levels of single-strand breaks in cells treated with 100 μmol/L MGd plus equimolar AA, as evidenced by changes in the alkaline tail moment (MGd + AA, 6 h: 14.7 ± 1.8; control: 2.8 ± 0.9). The level of single-strand breaks was dependent on the length of treatment. Motexafin gadolinium plus AA did not increase double-strand breaks. The repair of single-strand breaks at 2 h, but not at 4 h and 6 h, after irradiation was altered significantly in cells treated with MGd plus AA (MGd + AA, 2 h: 15.8 ± 3.4; control: 5.8 ± 0.6). Motexafin gadolinium did not alter the repair of double-strand breaks at any time after irradiation with 10 Gy. Conclusions: Motexafin gadolinium plus AA generated ROS, which in turn altered GSH homeostasis and induced DNA strand breaks. The MGd plus AA-mediated alteration of GSH levels increased the hypoxic, but not aerobic, radiosensitivity of EMT6 cells. Motexafin gadolinium altered the kinetics of single-strand break repair soon after irradiation but did not

  13. Studying the synergistic damage effects induced by 1.8 GHz radiofrequency field radiation (RFR) with four chemical mutagens on human lymphocyte DNA using comet assay in vitro.

    Science.gov (United States)

    Baohong, Wang; Jiliang, He; Lifen, Jin; Deqiang, Lu; Wei, Zheng; Jianlin, Lou; Hongping, Deng

    2005-10-15

    The aim of this investigation was to study the synergistic DNA damage effects in human lymphocytes induced by 1.8 GHz radiofrequency field radiation (RFR, SAR of 3 W/kg) with four chemical mutagens, i.e. mitomycin C (MMC, DNA crosslinker), bleomycin (BLM, radiomimetic agent), methyl methanesulfonate (MMS, alkylating agent), and 4-nitroquinoline-1-oxide (4NQO, UV-mimetic agent). The DNA damage of lymphocytes exposed to RFR and/or with chemical mutagens was detected at two incubation time (0 or 21 h) after treatment with comet assay in vitro. Three combinative exposure ways were used. Cells were exposed to RFR and chemical mutagens for 2 and 3h, respectively. Tail length (TL) and tail moment (TM) were utilized as DNA damage indexes. The results showed no difference of DNA damage indexes between RFR group and control group at 0 and 21 h incubation after exposure (P>0.05). There were significant difference of DNA damage indexes between MMC group and RFR+MMC co-exposure group at 0 and 21 h incubation after treatment (PRFR+4NQO co-exposure group at 0 and 21 h incubation after treatment was observed (PRFR+BLM co-exposure groups and RFR+MMS co-exposure groups was not significantly increased, as compared with corresponding BLM and MMS groups (P>0.05). The experimental results indicated 1.8 GHz RFR (SAR, 3 W/kg) for 2h did not induce the human lymphocyte DNA damage effects in vitro, but could enhance the human lymphocyte DNA damage effects induced by MMC and 4NQO. The synergistic DNA damage effects of 1.8 GHz RFR with BLM or MMS were not obvious.

  14. A study of DNA damage in buccal cells of consumers of well- and/or tap-water using the comet assay: Assessment of occupational exposure to genotoxicants.

    Science.gov (United States)

    Vazquez Boucard, Celia; Lee-Cruz, Larisa; Mercier, Laurence; Ramírez Orozco, Martín; Serrano Pinto, Vania; Anguiano, Gerardo; Cazares, Linette; Díaz, Daniel

    2017-10-01

    Because of concerns that natural aquifers in the region of Todos Santos (Baja California Sur, Mexico) might be contaminated by organochlorine pesticides and heavy metals, a case-control study was conducted among consumers and non-consumers of well- and/or tap-water to determine risks to human health. This study was based on a genotoxic evaluation of buccal cells using the Comet assay technique. Levels of DNA damage in the consumers group were significantly higher than those of the control group. However, occupational exposure to genotoxicants showed to be the critical factor rather than water consumption. Taking into account the professions of well- and/or tap-water consumers, agricultural workers exposed directly (those who fumigated) or indirectly (those not involved in fumigating) to agrochemicals showed greater genetic damage than controls. This difference persisted even when age, and whether the person smoked or consumed alcoholic drinks were considered. These factors were not associated with the level of genetic damage observed. Chemical analyses of organochlorine pesticides and heavy metals were carried out to evaluate the water quality of wells, faucets, and surface water of canals consumed by the population and/or used for irrigation. High concentrations of α and β endosulfan were detected in water of surface canals. Although our inventory of agrochemicals employed in the region showed the use of products considered carcinogenic and/or mutagenic, they were not detected by the analytical techniques used. Heavy metals (arsenic, mercury, and lead) were detected in water of some wells used for irrigation and human consumption. Environ. Mol. Mutagen. 58:619-627, 2017. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

  15. Genotoxicity effects of nano bioactive glass and Novabone bioglass on gingival fibroblasts using single cell gel electrophoresis (comet assay: An in vitro study

    Directory of Open Access Journals (Sweden)

    Mohammad Tavakoli

    2012-01-01

    Conclusion: The findings of this study have demonstrated that novel nano bioactive glass had no genotoxicity in concentrations lower than 4 mg/ml. Nanoparticles have a higher surface area in comparison to microparticles and thus, the amount and rate of ion release for nanoparticles are extremely higher. This difference is the main reason for the different genotoxicity of nano bioactive glass and micro Novabone bioglass in the concentrations higher than 4 mg/ml.

  16. Oligoclonal Pattern/Abnormal Protein Bands in Post-Treatment Plasma Cell Myeloma Patients: Implications for Protein Electrophoresis and Serum Free Light Chain Assay Results.

    Science.gov (United States)

    Singh, Gurmukh

    2017-08-01

    The impact of autologous stem cell transplantation (ASCT) in plasma cell myeloma patients on the frequency, quality, and timing of oligoclonal pattern in serum protein electrophoresis/immunofixation electrophoresis (SPEP/SIFE) and serum free light chain assay (SFLCA) was evaluated. Laboratory results and clinical data for 251 patients with plasma cell myeloma, who had SPEP/SIFE and/or SFLCA performed between January 2010 and December 2016 were reviewed. The results for SPEP/SIFE and SFLCA were compared in patients with ASCT to those without ASCT. The implications of oligoclonal pattern in interpretation of SPEP/SIFE and SFLCA - κ/λ ratio were addressed. In 251 patients, a total of 3,134 observations, of either SPEP/SIFE and/or SFLCA, were reviewed. One hundred fifty-nine patients received ASCT. The incidence of oligoclonal patterns was significantly higher after ASCT. More than half of the oligoclonal patterns developed in the first year after transplantation. In 13 of the 84 patients with lambda chain restricted plasma cell myeloma, the κ/λ ratio was kappa dominant in the presence of oligoclonal pattern. There was no reversal of κ/λ ratio in patients with kappa chain restricted plasma cell myelomas. ASCT is associated with significantly higher incidence of oligoclonal patterns than with chemotherapy alone. The presence of oligoclonal patterns has the potential to interfere with the interpretation of SPEP/SIFE and ascertainment of complete remission. At a minimum, the oligoclonal pattern caused an incorrect kappa dominant κ/λ ratio in 15.5% of patients with lambda chain restricted plasma cell myeloma. If a similar rate were to be applied to the 167 kappa chain myeloma patients, about 26 of these would have displayed an erroneous kappa chain dominant κ/λ ratio. The presence of oligoclonal pattern further degrades the performance of already dubious SFLCA. The need for recording the location of monoclonal spike in SPEP/SIFE and higher resolution protein

  17. Comparison of a direct enzymatic assay and polyacrylamide tube gel electrophoresis for measurement of small, dense low-density lipoprotein cholesterol.

    Science.gov (United States)

    Vanavanan, Somlak; Srisawasdi, Pornpen; Rochanawutanon, Mana; Kerdmongkol, Jirapa; Kroll, Martin H

    2015-01-01

    Small, dense low-density lipoprotein cholesterol (sdLDL-C) has been linked to the progression of cardiovascular disease. We compared two methods for determination of sdLDL-C, a direct enzymatic (ENZ) method and a polyacrylamide tube gel electrophoresis (PGE) assay, and investigated the associations of both sdLDL-C measurements with metabolic syndrome. We analyzed 242 patient sera for sdLDL and atherosclerosis-related markers. The PGE method separates the intermediate-density lipoprotein particles into three midbands (MID-A to MID-C) and the LDL particles into seven subfractions (LDL1 to LDL7); the sdLDL-PGE result is calculated as the sum of cholesterol concentrations from LDL3 to LDL7. The regression equation for sdLDL-C was [ENZmmol/L]=0.779[PGE]+0.67, r=0.713. ENZ showed higher sdLDL-C concentrations than PGE (0.86±0.33 vs. 0.24±0.32 mmol/L); however, the difference was not associated with sdLDL-C concentration (p=0.290). sdLDL-C, as measured with the enzymatic assay, exhibited significant positive correlations with very-low-density lipoprotein, MID-C, MID-B, and LDL2 (all p0.600). The ENZ and PGE methods yielded similar patterns of correlation between sdLDL-C, and atherosclerosis-related markers. Using logistic regression, sdLDL-ENZ and apolipoprotein B were identified as significant predictors of metabolic syndrome (p<0.03). The ENZ assay for sdLDL-C correlated well with the PGE method. The ENZ method measures a broader range of atherogenic lipoprotein particles than PGE and has the potential to identify subjects with vascular risk, thus contributing in directing specific interventions for cardiovascular prevention.

  18. Use of RNA amplification and electrophoresis for studying virus aerosol collection efficiency and their comparison with plaque assays.

    Science.gov (United States)

    Jiang, Xiao; Pan, Maohua; Hering, Susanne V; Lednicky, John A; Wu, Chang-Yu; Fan, Z Hugh

    2016-10-01

    The spread of virus-induced infectious diseases through airborne routes of transmission is a global concern for economic and medical reasons. To study virus transmission, it is essential to have an effective aerosol collector such as the growth tube collector (GTC) system that utilizes water-based condensation for collecting virus-containing aerosols. In this work, we characterized the GTC system using bacteriophage MS2 as a surrogate for a small RNA virus. We investigated using RNA extraction and reverse transcription- polymerase chain reaction (RT-PCR) to study the total virus collection efficiency of the GTC system. Plaque assays were also used to enumerate viable viruses collected by the GTC system compared to that by a commercially available apparatus, the SKC® Biosampler. The plaque assay counts were used to enumerate viable viruses whereas RT-PCR provides a total virus count, including those viruses inactivated during collection. The effects of relative humidity (RH) and other conditions on collection efficiency were also investigated. Our results suggest that the GTC has a collection efficiency for viable viruses between 0.24 and 1.8% and a total virus collection efficiency between 18.3 and 79.0%, which is 1-2 orders of magnitude higher than that of the SKC® Biosampler. Moreover, higher RH significantly increases both the viable and total collection efficiency of the GTC, while its effect on the collection efficiency of the SKC® Biosampler is not significant. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  19. Genotoxicity of carbon tetrachloride and the protective role of essential oil of Salvia officinalis L. in mice using chromosomal aberration, micronuclei formation, and comet assay.

    Science.gov (United States)

    Diab, Kawthar Ae; Fahmy, Maha A; Hassan, Zeinab M; Hassan, Emad M; Salama, Adel B; Omara, Enayat A

    2018-01-01

    The present work was conducted to evaluate the genotoxic effect of carbon tetrachloride (CCl 4 ) in mouse bone marrow and male germ cells. The safety and the modulating activity of sage (Salvia officinalis L.) essential oil (SEO) against the possible genotoxic effect of CCl 4 were also evaluated. A combination of in vivo mutagenic endpoints was included: micronucleus (MN), apoptosis using dual acridine orange/ethidium bromide (AO/EB) staining, comet assay, chromosomal aberrations (CAs), and sperm abnormalities. Histological examination of testis tissues was also studied. The extracted SEO was subjected to gas chromatography-mass spectrometry (GC-MS) for identifying its chemical constituents. Safety/genotoxicity of SEO was determined after two consecutive weeks (5 days/week) from oral treatment with different concentrations (0.1, 0.2, and 0.4 mL/kg). For assessing genotoxicity of CCl 4 , both acute (once) and subacute i.p. treatment for 2 weeks (3 days/week) with the concentrations 1.2 mL/kg (for acute) and 0.8 mL/kg (for subacute) were performed. For evaluating the protective role of SEO, simultaneous treatment with SEO plus CCl 4 was examined. In sperm abnormalities, mice were treated with the subject materials for five successive days and the samples were collected after 35 days from the beginning of treatment. Based on GC-MS findings, 22 components were identified in the chromatogram of SEO. The results demonstrated that the three concentrations of SEO were safe and non-genotoxic in all the tested endpoints. Negative results were also observed in bone marrow after acute and subacute treatment with CCl 4. In contrast, CCl 4 induced testicular DNA damage as evidenced by a significant increase of CAs in primary spermatocytes, sperm abnormalities, and histological distortion of testis. A remarkable reduction in these cells was observed in groups treated with SEO plus CCl 4 especially with the two higher concentrations of SEO. In conclusion, SEO is safe and

  20. 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.

  1. Whence comets?

    Science.gov (United States)

    A'Hearn, Michael F

    2006-12-15

    Recent advances in cometary science have indicated the importance of mixing of materials in the disk where the planets of our solar system formed. Now, the results from the Stardust Discovery Mission unambiguously show that even more extensive and earlier mixing of the material took place, raising new challenges for theories of the protoplanetary disk and the formation of comets.

  2. Biomonitoring of the genotoxic potential of aqueous extracts of soils and bottom ash resulting from municipal solid waste incineration, using the comet and micronucleus tests on amphibian (Xenopus laevis) larvae and bacterial assays (Mutatox and Ames tests).

    Science.gov (United States)

    Mouchet, F; Gauthier, L; Mailhes, C; Jourdain, M J; Ferrier, V; Triffault, G; Devaux, A

    2006-02-15

    The management of contaminated soils and wastes is a matter of considerable human concern. The present study evaluates the genotoxic potential of aqueous extracts of two soils (leachates) and of bottom ash resulting from municipal solid waste incineration (MSWIBA percolate), using amphibian larvae (Xenopus laevis). Soil A was contaminated by residues of solvents and metals and Soil B by polycyclic aromatic hydrocarbons and metals. MSWIBA was predominantly contaminated by metals. Two genotoxic endpoints were analysed in circulating erythrocytes taken from larvae: clastogenic and/or aneugenic effects (micronucleus induction) after 12 days of exposure and DNA-strand-breaking potency (comet assay) after 1 and 12 days of exposure. In addition, in vitro bacterial assays (Mutatox and Ames tests) were carried out and the results were compared with those of the amphibian test. Physicochemical analyses were also taken into account. Results obtained with the amphibians established the genotoxicity of the aqueous extracts and the comet assay revealed that they were genotoxic from the first day of exposure. The latter test could thus be considered as a genotoxicity-screening tool. Although genotoxicity persisted after 12 days' exposure, DNA damage decreased overall between days 1 and 12 in the MSWIBA percolate, in contrast to the soil leachates. Bacterial tests detected genotoxicity only for the leachate of soil A (Mutatox). The results confirm the ecotoxicological relevance of the amphibian model and underscore the importance of bioassays, as a complement to physico-chemical data, for risk evaluation.

  3. Genotoxicity and mutagenicity of water contaminated with tannery effluents, as evaluated by the micronucleus test and comet assay using the fish Oreochromis niloticus and chromosome aberrations in onion root-tips

    Directory of Open Access Journals (Sweden)

    Silvia Tamie Matsumoto

    2006-01-01

    Full Text Available Cytotoxicity of metals is important because some metals are potential mutagens able to induce tumors in humans and experimental animals. Chromium can damage DNA in several ways, including DNA double strand breaks (DSBs which generate chromosomal aberrations, micronucleus formation, sister chromatid exchange, formation of DNA adducts and alterations in DNA replication and transcription. In our study, water samples from three sites in the Córrego dos Bagres stream in the Franca municipality of the Brazilian state of São Paulo were subjected to the comet assay and micronucleus test using erythrocytes from the fish Oreochromis niloticus. Nuclear abnormalities of the erythrocytes included blebbed, notched and lobed nuclei, probably due to genotoxic chromium compounds. The greatest comet assay damage occurred with water from a chromium-containing tannery effluent discharge site, supporting the hypothesis that chromium residues can be genotoxic. The mutagenicity of the water samples was assessed using the onion root-tip cell assay, the most frequent chromosomal abnormalities observed being: c-metaphases, stick chromosome, chromosome breaks and losses, bridged anaphases, multipolar anaphases, and micronucleated and binucleated cells. Onion root-tip cell mutagenicity was highest for water samples containing the highest levels of chromium.

  4. Kinetics of DNA strand breaks and protection by antioxidants in UVA- or UVB-irradiated HaCaT keratinocytes using the single cell gel electrophoresis assay.

    Science.gov (United States)

    Lehmann, J; Pollet, D; Peker, S; Steinkraus, V; Hoppe, U

    1998-03-01

    The aim of this study was to characterize the genotoxic action of UVA and UVB in human keratinocytes by application of the single cell gel electrophoresis assay (SCGE assay). Dose dependence of DNA damage, the time course of its repair, and the influence of cellular antioxidant status were assessed. Irradiation with UVA or UVB both resulted in a dose-dependent increase in the level of DNA damage. A time course study to evaluate the repair kinetics in keratinocytes irradiated with 5 J/cm2 UVA revealed an immediate occurrence of DNA effects which subsequently disappeared within about 1 h, indicating removal of DNA lesions. This rapid repair of DNA damage is consistent with the observation that 5 J/cm2 UVA did not impair cellular viability. In contrast, exposure to 15 mJ/cm2 UVB resulted in a prolonged repair of DNA damage which lasted about 25 h. Thus, the repair kinetics of UVA- and UVB-induced DNA damage clearly differed from each other, implicating the induction of different types of DNA lesions by UVA and UVB. Neither a pretreatment with Mg-ascorbyl phosphate or D,L-alpha-tocopherol, nor depletion of endogenous glutathione altered cellular sensitivity to UVB. In contrast, the DNA damaging effects of UVA could be counteracted by a pretreatment with these antioxidants. These observations confirm that the UVA-induced effects on DNA are related to radical mediated strand breaks and DNA lesions forming alkali-labile sites. The UVB-induced effects mainly occur as a consequence of excision repair-related strand breaks. The observed repair kinetics of DNA lesions and the influence of cellular antioxidant status may help to elucidate protective mechanisms against the carcinogenic effects of UV radiation present in sunlight.

  5. Audit of Use and Overuse of Serum Protein Immunofixation Electrophoresis and Serum Free Light Chain Assay in Tertiary Health Care: A Case for Algorithmic Testing to Optimize Laboratory Utilization.

    Science.gov (United States)

    Heaton, Christopher; Vyas, Shikhar G; Singh, Gurmukh

    2016-04-01

    Overuse of laboratory tests is a persistent issue. We examined the use and overuse of serum immunofixation electrophoresis and serum free light chain assays to develop an algorithm for optimizing utilization. A retrospective review of all tests, for investigation of monoclonal gammopathies, for all patients who had any of these tests done from April 24, 2014, through July 25, 2014, was carried out. The test orders were categorized as warranted or not warranted according to criteria presented in the article. A total of 237 patients were tested, and their historical records included 1,503 episodes of testing for one or more of serum protein electrophoresis, serum immunofixation electrophoresis, and serum free light chain assays. Only 46% of the serum immunofixation and 42% serum free light chain assays were warranted. Proper utilization, at our institution alone, would have obviated $64,182.95/year in health care costs, reduced laboratory cost of reagent alone by $26,436.04/year, and put $21,904.92/year of part B reimbursement at risk. Fewer than half of the serum immunofixation and serum free light chain assays added value. The proposed algorithm for testing should improve utilization. Risk to part B billing may be a disincentive to reducing test utilization. © American Society for Clinical Pathology, 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  6. Bargain Electrophoresis.

    Science.gov (United States)

    Maderia, Vitor M. C.; Pires, Euclides M. V.

    1986-01-01

    Discusses the value of electrophoresis in the fields of protein chemistry and biochemistry. Describes how to build an inexpensive electrophoresis setup for use in either research or teaching activities. Details the construction of both the separating device and the power supply. (TW)

  7. Hemoglobin electrophoresis

    Science.gov (United States)

    ... page: //medlineplus.gov/ency/article/003639.htm Hemoglobin electrophoresis To use the sharing features on this page, please enable JavaScript. Hemoglobin is a protein that carries oxygen in the blood. Hemoglobin electrophoresis measures the levels of the different types of ...

  8. TCA precipitation and ethanol/HCl single-step purification evaluation: One-dimensional gel electrophoresis, bradford assays, spectrofluorometry and Raman spectroscopy data on HSA, Rnase, lysozyme - Mascots and Skyline data

    Directory of Open Access Journals (Sweden)

    Balkis Eddhif

    2018-04-01

    Full Text Available The data presented here are related to the research paper entitled “Study of a Novel Agent for TCA Precipitated Proteins Washing - Comprehensive Insights into the Role of Ethanol/HCl on Molten Globule State by Multi-Spectroscopic Analyses” (Eddhif et al., submitted for publication [1]. The suitability of ethanol/HCl for the washing of TCA-precipitated proteins was first investigated on standard solution of HSA, cellulase, ribonuclease and lysozyme. Recoveries were assessed by one-dimensional gel electrophoresis, Bradford assays and UPLC-HRMS. The mechanistic that triggers protein conformational changes at each purification stage was then investigated by Raman spectroscopy and spectrofluorometry. Finally, the efficiency of the method was evaluated on three different complex samples (mouse liver, river biofilm, loamy soil surface. Proteins profiling was assessed by gel electrophoresis and by UPLC-HRMS.

  9. Misleading presentation of haemoglobin electrophoresis data | Adu ...

    African Journals Online (AJOL)

    Haemoglobinopathies are common in sub-Saharan Africa. As such haemoglobin electrophoresis are required to inform clinical decision making. However, haemoglobin electrophoresis is an assay that detects protein at either alkaline or acidic pH. Such assays do not interrogate gene sequences but rather the product of a ...

  10. misleading presentation of haemoglobin electrophoresis data

    African Journals Online (AJOL)

    SUMMARY. Haemoglobinopathies are common in sub-Saharan Africa. As such haemoglobin electrophoresis are required to in- form clinical decision making. However, haemoglobin electrophoresis is an assay that detects protein at either alka- line or acidic pH. Such assays do not interrogate gene sequences but rather ...

  11. Transient DNA damage induced by high-frequency electromagnetic fields (GSM 1.8 GHz) in the human trophoblast HTR-8/SVneo cell line evaluated with the alkaline comet assay

    International Nuclear Information System (INIS)

    Franzellitti, Silvia; Valbonesi, Paola; Ciancaglini, Nicola; Biondi, Carla; Contin, Andrea; Bersani, Ferdinando; Fabbri, Elena

    2010-01-01

    One of the most controversial issue regarding high-frequency electromagnetic fields (HF-EMF) is their putative capacity to affect DNA integrity. This is of particular concern due to the increasing use of HF-EMF in communication technologies, including mobile phones. Although epidemiological studies report no detrimental effects on human health, the possible disturbance generated by HF-EMF on cell physiology remains controversial. In addition, the question remains as to whether cells are able to compensate their potential effects. We have previously reported that a 1-h exposure to amplitude-modulated 1.8 GHz sinusoidal waves (GSM-217 Hz, SAR = 2 W/kg) largely used in mobile telephony did not cause increased levels of primary DNA damage in human trophoblast HTR-8/SVneo cells. Nevertheless, further investigations on trophoblast cell responses after exposure to GSM signals of different types and durations were considered of interest. In the present work, HTR-8/SVneo cells were exposed for 4, 16 or 24 h to 1.8 GHz continuous wave (CW) and different GSM signals, namely GSM-217 Hz and GSM-Talk (intermittent exposure: 5 min field on, 10 min field off). The alkaline comet assay was used to evaluate primary DNA damages and/or strand breaks due to uncompleted repair processes in HF-EMF exposed samples. The amplitude-modulated signals GSM-217 Hz and GSM-Talk induced a significant increase in comet parameters in trophoblast cells after 16 and 24 h of exposure, while the un-modulated CW was ineffective. However, alterations were rapidly recovered and the DNA integrity of HF-EMF exposed cells was similar to that of sham-exposed cells within 2 h of recovery in the absence irradiation. Our data suggest that HF-EMF with a carrier frequency and modulation scheme typical of the GSM signal may affect the DNA integrity.

  12. Transient DNA damage induced by high-frequency electromagnetic fields (GSM 1.8 GHz) in the human trophoblast HTR-8/SVneo cell line evaluated with the alkaline comet assay.

    Science.gov (United States)

    Franzellitti, Silvia; Valbonesi, Paola; Ciancaglini, Nicola; Biondi, Carla; Contin, Andrea; Bersani, Ferdinando; Fabbri, Elena

    2010-01-05

    One of the most controversial issue regarding high-frequency electromagnetic fields (HF-EMF) is their putative capacity to affect DNA integrity. This is of particular concern due to the increasing use of HF-EMF in communication technologies, including mobile phones. Although epidemiological studies report no detrimental effects on human health, the possible disturbance generated by HF-EMF on cell physiology remains controversial. In addition, the question remains as to whether cells are able to compensate their potential effects. We have previously reported that a 1-h exposure to amplitude-modulated 1.8 GHz sinusoidal waves (GSM-217 Hz, SAR=2 W/kg) largely used in mobile telephony did not cause increased levels of primary DNA damage in human trophoblast HTR-8/SVneo cells. Nevertheless, further investigations on trophoblast cell responses after exposure to GSM signals of different types and durations were considered of interest. In the present work, HTR-8/SVneo cells were exposed for 4, 16 or 24h to 1.8 GHz continuous wave (CW) and different GSM signals, namely GSM-217 Hz and GSM-Talk (intermittent exposure: 5 min field on, 10 min field off). The alkaline comet assay was used to evaluate primary DNA damages and/or strand breaks due to uncompleted repair processes in HF-EMF exposed samples. The amplitude-modulated signals GSM-217 Hz and GSM-Talk induced a significant increase in comet parameters in trophoblast cells after 16 and 24h of exposure, while the un-modulated CW was ineffective. However, alterations were rapidly recovered and the DNA integrity of HF-EMF exposed cells was similar to that of sham-exposed cells within 2h of recovery in the absence irradiation. Our data suggest that HF-EMF with a carrier frequency and modulation scheme typical of the GSM signal may affect the DNA integrity.

  13. Application of the microbiological method DEFT/APC and DNA comet assay to detect ionizing radiation processing of minimally processed vegetables; Aplicacao do metodo microbiologico DEFT/APC e do teste do cometa na deteccao do tratamento com radiacao ionizante de hortalicas minimamente processadas

    Energy Technology Data Exchange (ETDEWEB)

    Araujo, Michel Mozeika

    2008-07-01

    Marketing of minimally processed vegetables (MPV) are gaining impetus due to its convenience, freshness and apparent healthy. However, minimal processing does not reduce pathogenic microorganisms to safe levels. Food irradiation is used to extend the shelf life and inactivation of food-borne pathogens, Its combination with minimal processing could improve the safety and quality of MPV. Two different food irradiation detection methods, a biological, the DEFT/APC, and another biochemical, the DNA Comet Assay were applied to MPV in order to test its applicability to detect irradiation treatment. DEFT/APC is a microbiological screening method based on the use of the direct epi fluorescent filter technique (DEFT) and the aerobic plate count (APC). DNA Comet Assay detects DNA damage due to ionizing radiation. Samples of lettuce, chard, watercress, dandelion, kale, chicory, spinach, cabbage from retail market were irradiated O.5 kGy and 1.0 kGy using a {sup 60} Co facility. Irradiation treatment guaranteed at least 2 log cycle reduction for aerobic and psychotropic microorganisms. In general, with increasing radiation doses, DEFT counts remained similar independent of irradiation processing while APC counts decreased gradually. The difference of the two counts gradually increased with dose increment in all samples. It could be suggested that a DEFT/APC difference over 2.0 log would be a criteria to judge if a MPV was treated by irradiation. DNA Comet Assay allowed distinguishing non-irradiated samples from irradiated ones, which showed different types of comets owing to DNA fragmentation. Both DEFT/APC method and DNA Comet Assay would be satisfactorily used as a screening method for indicating irradiation processing. (author)

  14. Monomorphism in humans and sequence differences among higher primates for a sequence tagged site (STS) in homeo box cluster 2 as assayed by denaturing gradient electrophoresis

    Energy Technology Data Exchange (ETDEWEB)

    Ruano, G.; Ruddle, F.H.; Kidd, K.K. (Yale Univ., New Haven, CT (United States)); Gray, M.R. (Tufts Univ., Boston, MA (United States)); Miki, Tetsuro (Osaka Univ. (Japan)); Ferguson-Smith, A.C. (Inst. of Animal Physiology and Genetics Research, Cambridge (United Kingdom))

    1990-03-11

    The human homeo box cluster 2 (HOX2) contains genes coding for DNA binding proteins involved in developmental control and is highly conserved between mouse and man. The authors have applied in concert the Polymerase Chain Reaction (PCR) and Denaturing Gradient Electrophoresis (DGE) to amplify defined primate HOX2 segments and to detect sequence differences among them. They have sequenced a PstI fragment 4 kb upstream from HOX 2.2 and synthesized primers delimiting both halves of 630 bp segment within it PCR on various unrelated humans and SC-PCR on chimpanzee, gorilla, orangutan and gibbon yielded products of the same length for each primer pair.

  15. Disintegration of comet nuclei

    Science.gov (United States)

    Ksanfomality, Leonid V.

    2012-02-01

    The breaking up of comets into separate pieces, each with its own tail, was seen many times by astronomers of the past. The phenomenon was in sharp contrast to the idea of the eternal and unchangeable celestial firmament and was commonly believed to be an omen of impending disaster, especially for comets with tails stretching across half the sky. It is only now that we have efficient enough space exploration tools to see comet nuclei and even - in the particular case of small comet Hartley-2 in 2010 - to watch their disintegration stage. There are also other suspected candidates for disintegration in the vast family of comet nuclei and other Solar System bodies.

  16. Physics of comets

    CERN Document Server

    Krishna Swamy, K S

    1997-01-01

    The study of Comet Halley in 1986 was a tremendous success for cometary science. In March of that year, six spacecrafts passed through Comet Halley as close as 600 km from the nucleus and made the in situ measurements of various kinds. These space missions to Comet Halley and that of the ICE spacecraft to Comet Giacobini-Zinner combined with studies, both ground-based and above the atmosphere, have increased our knowledge of cometary science in a dramatic way.This new edition of Physics of Comets incorporates these new and exciting findings. The emphasis of the book is on the physical processe

  17. Visually observing comets

    CERN Document Server

    Seargent, David A J

    2017-01-01

    In these days of computers and CCD cameras, visual comet observers can still contribute scientifically useful data with the help of this handy reference for use in the field. Comets are one of the principal areas for productive pro-amateur collaboration in astronomy, but finding comets requires a different approach than the observing of more predictable targets. Principally directed toward amateur astronomers who prefer visual observing or who are interested in discovering a new comet or visually monitoring the behavior of known comets, it includes all the advice needed to thrive as a comet observer. After presenting a brief overview of the nature of comets and how we came to the modern understanding of comets, this book details the various types of observations that can usefully be carried out at the eyepiece of a telescope. Subjects range from how to search for new comets to visually estimating the brightness of comets and the length and orientation of tails, in addition to what to look for in comet heads a...

  18. Study of DNA damage induction and repair capacity of fresh and cryopreserved lymphocytes exposed to H2O2 and gamma-irradiation with the alkaline comet assay.

    Science.gov (United States)

    Visvardis, E E; Tassiou, A M; Piperakis, S M

    1997-01-31

    The alkaline SCGE assay was evaluated for use with cryopreserved lymphocytes in order to obtain results similar to the freshly isolated ones. The induction of DNA damage as well as the repair capacity of gamma-rays and H2O2 exposed cryopreserved human lymphocytes was found to be the same to that of the freshly isolated. Human lymphocytes (fresh or cryopreserved) responded differently to the effects of gamma-irradiation if compared to the H2O2 treatment. The distribution of DNA damage among gamma-irradiated lymphocytes was more homogeneous compared to H2O2, both in freshly isolated and in cryopreserved cells. 2.4 micrograms/ml phytohemagglutinin at the start of a 2-h incubation in RPMI of cryopreserved samples gave similar DNA repair and distribution patterns to the 2-h post-exposure incubation of freshly isolated lymphocytes. H2O2-induced DNA damage was not repaired completely. However, the repair of gamma-rays-induced DNA damage was more efficient. These findings confirm the different mode of action of the two agents on the induction of DNA damage, as well as, the different response of the lymphocytes' DNA repair system.

  19. Ammonia abundances in comets

    Science.gov (United States)

    Wyckoff, S.; Tegler, S.; Engel, L.

    The emission band strengths of the NH2 bands of Comets Halley, Hartley-Good, Thiele, and Borrelly were measured to determine the NH2 column densities for the comets. Production rates obtained using the Haser and vectorial models are in agreement within the observational errors, suggesting that a simple two-step decay model may be used to approximate the NH2 distribution in a comet's coma. Ammonia-to-water abundance ratios from 0.01 to 0.4 percent were found for the four comets. The ratio in Comet Halley is found to be Q(NH3)/Q(H2O) = 0.002 + or - 0.001. No significant difference in the ammonia abundance was found before or after perihelion in Comet Halley.

  20. The pursuit of the genome instability by comet assay (single cell gel electrophoresis) in patients with cancer of the cavum in western Algerian; La recherche de l'instabilite genomique par le test des cometes (single cell gel electrophoresis) chez les malades atteints d'un cancer du cavum dans l'Ouest algerien

    Energy Technology Data Exchange (ETDEWEB)

    Boukerche, A.; Dali-Youcef, A.F. [Service de Radiotherapie, Oran (Algeria); Bouali-Youcef, Y. [Laboratoire d' Immunologie, Oran (Algeria); Mehadji, M. [Service d' ORL, Oran (Algeria); Chenal, C. [Rennes-1 Univ., UMR CNRS 6853, 35 (France)

    2007-11-15

    The analysis of results has shown a constitutional genome instability among the patients with a cavum cancer with a defect in DNA repair where some exogenous factors ( Epstein-Barr virus, EBV) seem play an important part. (N.C.)

  1. Special Report: Chemistry of Comets.

    Science.gov (United States)

    A'Hearn, Michael F.

    1984-01-01

    Discusses the chemistry of comets. How comets provide clues to the birth of the solar system, photolytic reactions on comets involving water, chemical modeling, nuclear chemistry, and research findings are among the areas considered. (JN)

  2. 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 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/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

  3. Evaluation of the internalization kinetics of the radiopharmaceutical 99mTc-N2S2-Tat(49-57)Lys3-Bn with diagnostic purposes, using comet assay

    International Nuclear Information System (INIS)

    Luna G, M. A.

    2011-01-01

    Gastrin-rea leasing peptide receptors (GRP-r) are over expressed in breast and prostate cancer cells. Bombesin (Bn) binds specifically and strongly to GRP-r and this is the base for to label the Bn with radionuclides by gamma rays. Tat (49-57) is a peptide that across the cell membrane easily so that, when it is conjugated to different proteins, it can works as a Trojan horse, facilitating the drug internalization to the cells. The radiopharmaceutical 99m Tc-N 2 S 2 -Tat(49-57)-Lys 3 -Bn was prepared for diagnosis and therapy at early stage of breast cancer. The objective of this study was to determine the role of Tat in the internalization kinetics of radiopharmaceuticals measured by DNA damage induced by means of comet assay. Human lymphocytes were treated with the following protocols: a) Tat-Bn, b) 99m Tc-Bn, or c) 99m Tc-N 2 S 2 -Tat(49-57)-Lys 3 -Bn, also an untreated group was conformed. The internalization was evaluated at 0, 5, 10, 15, 30 and 60 min after exposure with three repetitions each one, and for radiopharmaceuticals with 2.9, 6.6, 9.0 and 14.8 MBq activities. DNA damage was scored in 100 cells per time and treatment, as tail length and tail moment. A Kruskal-Wallis variance analysis with p≤ 0.05 was applied for comparison between treatments. The results showed that the damage caused by 99m Tc-N 2 S 2 -Tat(49-57)-Lys 3 -Bn is significantly higher than that caused by 99m Tc-Bn and Tat-Bn, showing that Tat favors the internalization of the radiopharmaceutical. (Author)

  4. Comet assay analysis of repair of DNA strand breaks in normal and deficient human cells exposed to radiations and chemicals. Evidence for a repair pathway specificity of DNA ligation

    Energy Technology Data Exchange (ETDEWEB)

    Nocentini, S. [Institut Curie de Biologie, Paris (France)

    1995-11-01

    The induction and resealing of DNA strand breaks in a cell line with a proven defect in DNA ligase I, 46BR, and in two Bloom`s syndrome cell lines. YBL6 and GM 1492, were compared to those observed in normal human 1BR/3 fibroblasts after treatment with a variety of genotoxic agents whose lesions are processed by different repair pathways. This analysis was performed using the single-cell gel electrophoresis assay. The three types of cells were found to have similar capabilities to recognize and incise ultraviolet photoproducts and also demonstrated similar amounts of DNA breaks immediately after {gamma} irradiation. During post-treatment incubation, 46BR cells showed a marked DNA re-ligation defect after ultraviolet radiation damage, GM 1492 cells demonstrated a highly reduced DNA joining ability after relatively high doses of ultraviolet radiation, and YBL6 cells were particularly affected in DNA re-ligation after damage by 4-nitroquinoline-1-oxide. The two Bloom`s syndrome cell lines and 46BR cells had a nearly normal ability to reseal breaks resulting from {gamma} irradiation or treatment with xanthine plus xanthine oxidase. These findings suggest that different DNA ligases may be involved in different DNA repair pathways in human cells. 60 refs., 7 figs.

  5. Changing Speed of Comets

    Science.gov (United States)

    Follows, Mike

    2003-01-01

    It is shown that highly elliptical orbits, such as those of comets, can be explained well in terms of energy rather than forces. The principle of conservation of energy allows a comet's velocity to be calculated at aphelion and perihelion. An example asks students to calculate whether they can run fast enough to escape from a small asteroid.…

  6. Radiobiological study on DNA strand breaks and repair using single cell gel electrophoresis

    International Nuclear Information System (INIS)

    Ikushima, Takaji

    1994-01-01

    Single cell gel electrophoresis (SCGE) provides a novel method to measure DNA damage in individual cells and more importantly, to assess heterogeneity in response within a mixed population of cells. Cells embedded in agarose are lysed, subjected to electrophoresis, stained with a fluorescent DNA-specific dye, and viewed under a fluorescence microscope. Damaged cells display 'comets', broken DNA migrating farther to the anode in the electric field. We have previously used this technique to quantify DNA damage induced by moderate doses of low and high LET radiations in cultured Chinese hamster cells. The assay has been optimized in terms of lysing and electrophoresis conditions, and applied to analyse the DNA strand breaks, their repair kinetics and heterogeneity in response in individual Chinese hamster cells exposed to gamma-rays, and to KUR thermal neutrons with and without 10 B or to KEK PF monochromatic soft X-rays as well as to a radio-mimetic agent, neocarzinostatin. The DNA double-strand breaks induced by boron-neutron captured reactions were repaired at a slower rate, but a heterogeneity in response might not contribute to the difference. The neocarzinostatin-induced DNA damage were efficiently repaired in a dose-dependent fashion. The initial amount of gamma-ray induced DNA double-strand breaks was not significantly altered in cells pre-exposed to very low adapting dose. (author)

  7. DNA Comet Giemsa Staining for Conventional Bright-Field Microscopy

    Directory of Open Access Journals (Sweden)

    Andreyan Osipov

    2014-04-01

    Full Text Available This study was undertaken to evaluate the compatibility of Giemsa staining protocol with the comet assay. We showed, for the first time, that DNA comets can be visualized and analyzed using Giemsa staining. We generated DNA damage dose response curves for human peripheral blood lymphocytes exposed to X-ray radiation using the comet assay with either SybrGreen I or Giemsa stain. The dose response curves were fitted by linear regressions (R2 > 0.977. The SybrGreen I results showed only ~1.2-fold higher slope coefficient (method sensitivity compared to the Giemsa results. The unexpectedly high sensitivity of Giemsa staining for the comet assay is due to the Romanowsky-Giemsa effect, the stain photo-stability and the higher resolution of bright-field imaging compared to fluorescence imaging. Our results demonstrate that Giemsa staining can effectively be used for measuring DNA damage by the comet assay. The low cost and availability of Giemsa stain makes this method affordable for any low budget research and will facilitate new applications of the comet assay in biology and medicine.

  8. DNA Comet Giemsa Staining for Conventional Bright-Field Microscopy

    Science.gov (United States)

    Osipov, Andreyan; Arkhangelskaya, Ekaterina; Vinokurov, Alexei; Smetanina, Nadezhda; Zhavoronkov, Alex; Klokov, Dmitry

    2014-01-01

    This study was undertaken to evaluate the compatibility of Giemsa staining protocol with the comet assay. We showed, for the first time, that DNA comets can be visualized and analyzed using Giemsa staining. We generated DNA damage dose response curves for human peripheral blood lymphocytes exposed to X-ray radiation using the comet assay with either SybrGreen I or Giemsa stain. The dose response curves were fitted by linear regressions (R2 > 0.977). The SybrGreen I results showed only ~1.2-fold higher slope coefficient (method sensitivity) compared to the Giemsa results. The unexpectedly high sensitivity of Giemsa staining for the comet assay is due to the Romanowsky-Giemsa effect, the stain photo-stability and the higher resolution of bright-field imaging compared to fluorescence imaging. Our results demonstrate that Giemsa staining can effectively be used for measuring DNA damage by the comet assay. The low cost and availability of Giemsa stain makes this method affordable for any low budget research and will facilitate new applications of the comet assay in biology and medicine. PMID:24727376

  9. Application of single cell gel electrophoresis in post-evaluation of organism radiation damage

    International Nuclear Information System (INIS)

    Jiang Lin; Mu Wanjun; Liu Guoping; Xu Yunshu; Luo Shunzhong; Gao Qingxiang

    2009-01-01

    The transient irradiation-caused DNA damage in the human peripheral blood lymphocytes,mouse peripheral blood lymphocytes and alive mouse irradiated by α-ray and γ-ray was investigated, and the single cell gel electrophoresis(SCGE, Comet Assay) was used to detect the extent of DNA damage. On this basis, the dose-effect curve and the evaluating method for radiant after-effect were established, the absorbed dose of alive mouse A irradiated by γ-rays was computed. The results indicate that not only the dose-effect can be described by using SCGE, but also the dose-computed after organism irradiated by radiant rays is achieved with it, and SCGE may be used as a new biological dosimeter. (authors)

  10. Application of single cell gel electrophoresis in post-evaluation of organism radiation damage

    International Nuclear Information System (INIS)

    Jiang Lin; Mu Wanjun; Liu Guoping; Xu Yunshu; Gao Qingxiang

    2007-01-01

    The immediate irradiation-caused DNA damage in the human Peripheral Blood Lymphocytes, mouse Peripheral Blood Lymphocytes and alive mouse irradiated by α-Rays and γ-rays was investigated, and the single cell gel electrophoresis(SCGE, Comet Assay) was used to detect the extent of DNA damage. On this base, the dose-effect curve and the evaluating method for radiant aftereffect were established, the absorbed does of alive mouse A irradiated by γ-rays was computed. The results indicated that not only the does-effect could be described by using SCGE, but also the does-computed after organism irradiated by radiant rays was achieved with it, and SCGE might be used as a new biological dosimeter. (authors)

  11. Realm of the comets

    International Nuclear Information System (INIS)

    Weissman, P.R.

    1987-01-01

    Studies of Jovian perturbations of the orbits of long-period comets led to the concept of the Oort cloud of 180 billion comets at 50,000-150,000 AU from the sun. Several comets are induced to move toward the sun every million years by the passage of a star at a distance of a few light years. The location of the cloud has since been revised to 20,000-100,000 AU, and comets are now accepted as remnant material fron the proto-solar system epoch. The galactic disk and random, close-passing stars may also cause rare, large perturbations in the orbits of the cloud comets, sending large numbers of comets through the inner solar system. The resulting cometary storm is a candidate cause for the wholesale extinction of dinosaurs in the Cretaceous-Terniary transition due to large number of planetesimals, or one large comet, striking the earth, in a short period of time. The IRAS instruments have detected similar clouds of material around other stars

  12. Comets in UV

    Science.gov (United States)

    Shustov, B.; Sachkov, M.; Gómez de Castro, A. I.; Vallejo, J. C.; Kanev, E.; Dorofeeva, V.

    2018-04-01

    Comets are important "eyewitnesses" of Solar System formation and evolution. Important tests to determine the chemical composition and to study the physical processes in cometary nuclei and coma need data in the UV range of the electromagnetic spectrum. Comprehensive and complete studies require additional ground-based observations and in situ experiments. We briefly review observations of comets in the ultraviolet (UV) and discuss the prospects of UV observations of comets and exocomets with space-borne instruments. A special reference is made to the World Space Observatory-Ultraviolet (WSO-UV) project.

  13. Comets: looking ahead.

    Science.gov (United States)

    A'Hearn, Michael F

    2017-07-13

    We outline the key questions about comets that must be answered in order to understand cometary formation in the context of the protoplanetary disc and the role of comets in the formation and evolution of the solar system. We then discuss the new understanding of comets from Rosetta and from other recent advances, including work presented by others at the discussion meeting. Finally, we suggest some key directions for future projects to better address the above questions.This article is part of the themed issue 'Cometary science after Rosetta'. © 2017 The Author(s).

  14. DIRBE Comet Trails

    Science.gov (United States)

    Arendt, Richard G.

    2015-01-01

    Re-examination of the COBE DIRBE data reveals the thermal emission of several comet dust trails.The dust trails of 1P/Halley, 169P/NEAT, and 3200 Phaethon have not been previously reported.The known trails of 2P/Encke, and 73P/Schwassmann-Wachmann 3 are also seen. The dust trails have 12 and 25 microns surface brightnesses of comet, but others are best seen at high ecliptic latitudes as the Earth passes over or under the dust trail. All these comets have known associations with meteor showers. This re-examination also reveals one additional comet and 13 additional asteroids that had not previously been recognized in the DIRBE data.

  15. William Herschel and Comets

    Science.gov (United States)

    Sullivan, Woodruff

    2018-01-01

    I examine the observational and theoretical researches of William Herschel on 21 comets that he observed over the period 1781 to 1812. Herschel's focus, unlike most contemporaries, was on their physical structure, not their orbits. He forged a strong connection between comets and his nebulae with a scheme of cometary "maturation" (1812) involved a comet traveling from star to star after its central "planetary body'; was born from gravitational collapse of a nebula. During close passages of a star, the comet brightened and lost mass from its atmosphere; at other times, when between stars, it encountered nebulae and was rejuvenated by picking up more mass. Laplace soon adopted these ideas to improve his nebula hypothesis for solar system formation.

  16. Genotoxicity of three food processing contaminants in transgenic mice expressing human sulfotransferases 1A1 and 1A2 as assessed by the in vivo alkaline single cell gel electrophoresis assay.

    Science.gov (United States)

    Høie, Anja Hortemo; Svendsen, Camilla; Brunborg, Gunnar; Glatt, Hansruedi; Alexander, Jan; Meinl, Walter; Husøy, Trine

    2015-10-01

    The food processing contaminants 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), 5-hydroxymethylfurfural (HMF) and 2,5 dimethylfuran (DMF) are potentially both mutagenic and carcinogenic in vitro and/or in vivo, although data on DMF is lacking. The PHIP metabolite N-hydroxy-PhIP and HMF are bioactivated by sulfotransferases (SULTs). The substrate specificity and tissue distribution of SULTs differs between species. A single oral dose of PhIP, HMF or DMF was administered to wild-type (wt) mice and mice expressing human SULT1A1/1A2 (hSULT mice). DNA damage was studied using the in vivo alkaline single cell gel electrophoresis (SCGE) assay. No effects were detected in wt mice. In the hSULT mice, PhIP and HMF exposure increased the levels of DNA damage in the liver and kidney, respectively. DMF was not found to be genotoxic. The observation of increased DNA damage in hSULT mice compared with wt mice supports the role of human SULTs in the bioactivation of N-hydroxy-PhIP and HMF in vivo. © 2015 The Authors. Environmental and Molecular Mutagenesis Published by Wiley Periodicals, Inc.

  17. Localization of specific sequences and DNA single-strand breaks in individual UV-A-irradiated human lymphocytes by COMET FISH

    Science.gov (United States)

    Bock, Claudia; Rapp, Alexander; Dittmar, Heike; Monajembashi, Shamci; Greulich, Karl-Otto

    1999-01-01

    The COMET assay, a single cell electrophoresis technique which allows to separate electrophoretically fractionated DNA according to size has been combined with fluorescence in situ hybridization (FISH) which allows to localize specific genes or gene regions. This combination (COMET FISH) allows the detection of DNA single strand breaks in specific regions of the genome of human lymphocytes at the single cell level. Various types of DNA probes, e.g. centromere-, (alpha) - satellite-, telomere-, whole chromosome-, single copy- and region specific DNA probes have been used to investigate whether the UV-A induced DNA single strand breaks are distributed randomly all over the human genome or induced at specific sites ('hot spots'). In the investigated human peripheral blood lymphocytes all but one centromere reveal low sensitivity for UV-A irradiation (500 kJ/m2), while telomeres are randomly distributed over COMET heads and tails. The human chromosome 1 is fractionated by irradiation, but remains in the COMET head, indicating an only moderate degree of fractionation. Among three tested single copy probes, c- myc, p53 and p58, the p53 gene located on chromosome 17p13.1 and the p58 gene (1p36) appear to be located in UV-A stable regions of the human genome in 95% of 65 investigated lymphocytes. In contrast, the c-myc proto-oncogene (8q24) is found in the COMET tail in 90% of the 27 investigated lymphocytes and thus appears to be more sensitive to UV-A irradiation.

  18. Comets in Australian Aboriginal Astronomy

    Science.gov (United States)

    Hamacher, Duane W.; Norris, Ray P.

    2011-03-01

    We present 25 accounts of comets from 40 Australian Aboriginal communities, citing both supernatural perceptions of comets and historical accounts of historically bright comets. Historical and ethnographic descriptions include the Great Comets of 1843, 1861, 1901, 1910, and 1927. We describe the perceptions of comets in Aboriginal societies and show that they are typically associated with fear, death, omens, malevolent spirits, and evil magic, consistent with many cultures around the world. We also provide a list of words for comets in 16 different Aboriginal languages.

  19. Protein electrophoresis - urine

    Science.gov (United States)

    ... medlineplus.gov/ency/article/003589.htm Urine protein electrophoresis test To use the sharing features on this page, please enable JavaScript. The urine protein electrophoresis (UPEP) test is used to estimate how much ...

  20. Serum globulin electrophoresis

    Science.gov (United States)

    ... medlineplus.gov/ency/article/003544.htm Serum globulin electrophoresis To use the sharing features on this page, please enable JavaScript. The serum globulin electrophoresis test measures the levels of proteins called globulins ...

  1. Jupiter Laser Facility - COMET Laser

    Data.gov (United States)

    Federal Laboratory Consortium — COMET has 4 beam configurations with uncompressed pulse lengths from 500 ps to 6 ns, compressed pulses to 0.5 ps, and beam energies up to 20 J. COMET can fire every...

  2. Discovering the Nature of Comets.

    Science.gov (United States)

    Whipple, Fred L.

    1986-01-01

    "The Mystery of Comets" by Dr. Fred Whipple provides an introduction to the modern picture of comets and his personal reminiscences of how his model of comets came to be. An adaptation of several sections of the book is presented. (JN)

  3. Term Projects on Interstellar Comets

    Science.gov (United States)

    Mack, John E.

    1975-01-01

    Presents two calculations of the probability of detection of an interstellar comet, under the hypothesis that such comets would escape from comet clouds similar to that believed to surround the sun. Proposes three problems, each of which would be a reasonable term project for a motivated undergraduate. (Author/MLH)

  4. DNA staining with the fluorochromes EtBr, DAPI and YOYO-1 in the comet assay with tobacco plants after treatment with ethyl methanesulphonate, hyperthermia and DNase-I

    Czech Academy of Sciences Publication Activity Database

    Gichner, Tomáš; Mukherjee, A.; Velemínský, Jiří

    2006-01-01

    Roč. 605, 1-2 (2006), s. 17-21 ISSN 1383-5718 R&D Projects: GA ČR GA521/05/0500 Institutional research plan: CEZ:AV0Z50380511 Keywords : DNA migration * Nicotiana tabacum * Single-cell gel electrophoresis Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.122, year: 2006

  5. Death of a comet

    CERN Multimedia

    Hawkes, N

    2000-01-01

    The comet Linear dissolved as it made its closest approach to the sun on July 25th. The first stages of its breakup had been witnessed by the Hubble telescope when it threw off a piece of its crust (3 paragraphs).

  6. Comet radar explorer

    Science.gov (United States)

    Farnham, Tony; Asphaug, Erik; Barucci, Antonella; Belton, Mike; Bockelee-Morvan, Dominique; Brownlee, Donald; Capria, Maria Teresa; Carter, Lynn; Chesley, Steve; Farnham, Tony; Gaskell, Robert; Gim, Young; Heggy, Essam; Herique, Alain; Klaasen, Ken; Kofman, Wlodek; Kreslavsky, Misha; Lisse, Casey; Orosei, Roberto; Plaut, Jeff; Scheeres, Dan

    The Comet Radar Explorer (CORE) is designed to perform a comprehensive and detailed exploration of the interior, surface, and inner coma structures of a scientifically impor-tant Jupiter family comet. These structures will be used to investigate the origins of cometary nuclei, their physical and geological evolution, and the mechanisms driving their spectacular activity. CORE is a high heritage spacecraft, injected by solar electric propulsion into orbit around a comet. It is capable of coherent deep radar imaging at decameter wavelengths, high resolution stereo color imaging, and near-IR imaging spectroscopy. Its primary objective is to obtain a high-resolution map of the interior structure of a comet nucleus at a resolution of ¿100 elements across the diameter. This structure shall be related to the surface geology and morphology, and to the structural details of the coma proximal to the nucleus. This is an ideal complement to the science from recent comet missions, providing insight into how comets work. Knowing the structure of the interior of a comet-what's inside-and how cometary activity works, is required before we can understand the requirements for a cryogenic sample return mission. But more than that, CORE is fundamental to understanding the origin of comets and their evolution in time. The mission is made feasible at low cost by the use of now-standard MARSIS-SHARAD reflec-tion radar imaging hardware and data processing, together with proven flight heritage of solar electric propulsion. Radar flight heritage has been demonstrated by the MARSIS radar on Mars Express (Picardi et al., Science 2005; Plaut et al., Science 2007), the SHARAD radar onboard the Mars Reconnaissance Orbiter (Seu et al., JGR 2007), and the LRS radar onboard Kaguya (Ono et al, EPS 2007). These instruments have discovered detailed subsurface structure to depths of several kilometers in a variety of terrains on Mars and the Moon. A reflection radar deployed in orbit about a comet

  7. Comets in Indian Scriptures

    Science.gov (United States)

    Das Gupta, P.

    2016-01-01

    The Indo-Aryans of ancient India observed stars and constellations for ascertaining auspicious times in order to conduct sacrificial rites ordained by the Vedas. Naturally, they would have sighted comets and referred to them in the Vedic texts. In Rigveda (circa 1700-1500 BC) and Atharvaveda (circa 1150 BC), there are references to dhumaketus and ketus, which stand for comets in Sanskrit. Rigveda speaks of a fig tree whose aerial roots spread out in the sky (Parpola 2010). Had this imagery been inspired by the resemblance of a comet's tail with long and linear roots of a banyan tree (ficus benghalensis)? Varahamihira (AD 550) and Ballal Sena (circa AD 1100-1200) described a large number of comets recorded by ancient seers, such as Parashara, Vriddha Garga, Narada, and Garga, to name a few. In this article, we propose that an episode in Mahabharata in which a radiant king, Nahusha, who rules the heavens and later turns into a serpent after he kicked the seer Agastya (also the star Canopus), is a mythological retelling of a cometary event.

  8. Halley's Comet: A Bibliography.

    Science.gov (United States)

    Freitag, Ruth S., Comp.

    Included in this bibliography are over 3,200 references to publications on Halley's Comet, its history, orbital motion, and physical characteristics, meteor streams associated with it, preparations for space missions to study it in 1986, and popular reaction to its appearances. Also cited are a few papers that, although they devote little…

  9. Splitting of Comets

    Indian Academy of Sciences (India)

    Home; Journals; Resonance – Journal of Science Education; Volume 7; Issue 1. Splitting of Comets. Utpal Mukhopadhyay. General Article Volume 7 Issue 1 January 2002 pp 11-22. Fulltext. Click here to view fulltext PDF. Permanent link: http://www.ias.ac.in/article/fulltext/reso/007/01/0011-0022. Keywords. Cometary ...

  10. Comet 2001 Q2

    Czech Academy of Sciences Publication Activity Database

    Pravec, Petr; Kušnirák, Peter; Bouma, R. J.; Raymundo, P. M.

    č. 7687 (2001), s. 1 ISSN 0081-0304 R&D Projects: GA ČR GA205/99/0255 Institutional research plan: CEZ:AV0Z1003909 Keywords : comets * astrometry Subject RIV: BN - Astronomy, Celestial Mechanics, Astrophysics

  11. Asteroids, meteorites, and comets

    CERN Document Server

    Elkins-Tanton, Linda T

    2010-01-01

    Asteroids, Comets, and Meteorites provides students, researchers, and general readers with the most up-to-date information on this fascinating field. From the days of the dinosaurs to our modern environment, this book explores all aspects of these cosmic invaders.

  12. DRBE comet trails

    Energy Technology Data Exchange (ETDEWEB)

    Arendt, Richard G., E-mail: Richard.G.Arendt@nasa.gov [CREST/UMBC, Code 665, NASA/GSFC, Greenbelt, MD 20771 (United States)

    2014-12-01

    Re-examination of the Cosmic Background Explorer Diffuse Infrared Background Experiment (DIRBE) data reveals the thermal emission of several comet dust trails. The dust trails of 1P/Halley, 169P/NEAT, and 3200 Phaethon have not been previously reported. The known trails of 2P/Encke and 73P/Schwassmann–Wachmann 3 are also seen. The dust trails have 12 and 25 μm surface brightnesses of <0.1 and <0.15 MJy sr{sup −1}, respectively, which is <1% of the zodiacal light intensity. The trails are very difficult to see in any single daily image of the sky, but are evident as rapidly moving linear features in movies of the DIRBE data. Some trails are clearest when crossing through the orbital plane of the parent comet, but others are best seen at high ecliptic latitudes as the Earth passes over or under the dust trail. All these comets have known associations with meteor showers. This re-examination also reveals 1 additional comet and 13 additional asteroids that had not previously been recognized in the DIRBE data.

  13. A Search of Reactivated Comets

    Science.gov (United States)

    Ye, Quan-Zhi

    2017-05-01

    Dormant or near-dormant short-period comets can unexpectedly regain the ability to eject dust. In many known cases, the resurrection is short-lived and lasts less than one orbit. However, it is possible that some resurrected comets can remain active in later perihelion passages. We search the archival images of various facilities to look for these “reactivated” comets. We identify two candidates, 297P/Beshore and 332P/Ikeya-Murakami, both of which were found to be inactive or weakly active in the previous orbit before their discovery. We derive a reactivation rate of ˜ 0.007 {{comet}}-1 {{orbit}}-1, which implies that typical short-period comets only become temporarily dormant a few times or less. Smaller comets are prone to rotational instability and may undergo temporary dormancy more frequently. Next generation high-cadence surveys may find more reactivation events of these comets.

  14. Cometography a catalog of comets

    CERN Document Server

    Kronk, Gary W; Seargent, David A J

    2017-01-01

    Cometography is a multi-volume catalog of every comet observed from ancient times up to the 1990s, when the internet took off as a medium of scientific record. It uses the most reliable orbits known to determine the distances from the Earth and Sun at the time of discovery and last observation, as well as the largest and smallest angular distance to the Sun, most northerly and southerly declination, closest distance to the Earth, and other details, to enable the reader to understand each comet's physical appearance. Volume 6, the final volume in the catalog, covers the observations and pertinent calculations for every comet seen between 1983 and 1993. The comets are listed in chronological order, with complete references to publications relating to each comet and physical descriptions of each comet's development throughout its apparition. Cometography is the definitive reference on comets through the ages, for astronomers and historians of science.

  15. Radiosensitivity evaluation of Human tumor cell lines by single cell gel electrophoresis

    International Nuclear Information System (INIS)

    Zhang Yipei; Cao Jia; Wang Yan; Du Liqing; Li Jin; Wang Qin; Fan Feiyue; Liu Qiang

    2011-01-01

    Objective: To explore the feasibility of determining radiosensitivity of human tumor cell lines in vitro using single cell gel electrophoresis (SCGE). Methods: Three human tumor cell lines were selected in this study, HepG 2 , EC-9706 and MCF-7. The surviving fraction (SF) and DNA damage were detected by MTT assay, nested PCR technique and comet assay respectively. Results: MTT assay: The SF of HepG 2 and EC-9706 after irradiated by 2, 4 and 8 Gy was lower significantly than that of MCF-7, which showed that the radiosensitivity of HepG 2 and EC-9706 was higher than that of MCF-7. But there was no statistical difference of SF between HepG 2 and EC-9706. SCGE: The difference of radiosensitivity among these three tumor cell lines was significant after 8 Gy γ-ray irradiation. Conclusion: The multi-utilization of many biological parameter is hopeful to evaluate the radiosensitivity of tumor cells more objectively and exactly. (authors)

  16. Automatic analysis of silver-stained comets by CellProfiler software.

    Science.gov (United States)

    González, J E; Romero, I; Barquinero, J F; García, O

    2012-10-09

    The comet assay is one of the most widely used methods to evaluate DNA damage and repair in eukaryotic cells. The comets can be measured by software, in a semi-automatic or automatic process. In this paper, we apply the CellProfiler open-source software for automatic analysis of comets from digitized images, reporting the percentage of tail DNA. A side-by-side comparison of CellProfiler with CASP software demonstrated good agreement between the two packages. Our work demonstrates that automatic measurement of silver-stained comets with open-source software is possible, providing significant time savings. © 2012 Elsevier B.V. All rights reserved.

  17. 67P, Singing Comet

    Science.gov (United States)

    Smirnova, Ekaterina

    2017-04-01

    I would like to propose to present a short science-art-music collaboration film called "67P, Singing Comet" (5:27 min). If time of the session will allow, prior to the film I would like to make a slide show introduction to this project, highlighting the inspiration - the mission Rosetta by the European Space Agency (ESA) - and the artistic collaboration that took place in creating this piece. Inspired by the ESA Rosetta mission to the comet 67P, Ekaterina Smirnova (artist and project director, New York), Lee Mottram (clarinetist, Wales), Takuto Fukuda (composer, Japan) and Brian Hekker (video editor, New York) collaborated to create a unique atmospheric piece. Water and the origins of life throughout the Universe (specifically the Earth) is an element of the mission and the focus of Ekaterina's artistic vision. Ekaterina literally and figuratively paints a sensory assemblage using a combination of synthetic and natural elements to shape this artistic creation. To paint her watercolor works she is using a replica of the water found on the comet and implementing her own heartbeat into the music to create a recognizable inward sound of life. The Electro-Acoustic composition by Takuto Fukuda features an electronically manipulated performance by clarinetist Lee Mottram. The piece ceremoniously begins with reverberant bursts of low-register atonal bells transporting the listener to their ethereal inner origins of body and mind. The imagination takes the experience to an unknown destination as it gains speed gliding through the visual and audible textures of space and time. The comet's water similarly reacts with an ebb and flow thawing ice to potentially give life a chance as it is thrust along an orbit around the Sun. Near then far from the heat the comet forms frozen particles from vapors as it reaches it's furthest stretches creating an aerodynamic tail of icicles that slowly dissipate in a cycle that repeats itself until the comet's ultimate collision with an

  18. Detecting irradiation of seeds using microgel electrophoresis (a collaborative trial)

    International Nuclear Information System (INIS)

    Cerda, H.; Haine, H.E.; Jones, J.L.

    1995-06-01

    Preservation of certain foods by irradiation is permitted in the United Kingdom. However, all irradiated foods must be labelled as such, to ensure consumer choice. To help enforce labelling, a variety of methods have been developed for distinguishing between irradiated and non-irradiated foods. In preliminary trials, microgel electrophoresis -a simple method of assessing DNA damage - has shown considerable promise in this respect. This report describes microgel electrophoresis, and details results obtained in a blind trial carried out in collaboration with the Swedish University of Agricultural Sciences. Microgel electrophoresis facilitates analysis of the leakage of DNA from cells extracted from food material. In irradiated samples, the DNA is fragmented and will leak from cells in an electric current. This leakage can be seen as a 'comet' when the stained gel is viewed with a microscope. The size and shape of the comet can be used to estimate the irradiation dose administered to the sample. In non-irradiated samples the DNA is less fragmented, will tend not to leak from the cells and will not form a comet. (author)

  19. CINE: Comet INfrared Excitation

    Science.gov (United States)

    de Val-Borro, Miguel; Cordiner, Martin A.; Milam, Stefanie N.; Charnley, Steven B.

    2017-08-01

    CINE calculates infrared pumping efficiencies that can be applied to the most common molecules found in cometary comae such as water, hydrogen cyanide or methanol. One of the main mechanisms for molecular excitation in comets is the fluorescence by the solar radiation followed by radiative decay to the ground vibrational state. This command-line tool calculates the effective pumping rates for rotational levels in the ground vibrational state scaled by the heliocentric distance of the comet. Fluorescence coefficients are useful for modeling rotational emission lines observed in cometary spectra at sub-millimeter wavelengths. Combined with computational methods to solve the radiative transfer equations based, e.g., on the Monte Carlo algorithm, this model can retrieve production rates and rotational temperatures from the observed emission spectrum.

  20. Diatoms in comets

    Science.gov (United States)

    Hoover, R.; Hoyle, F.; Wallis, M. K.; Wickramasinghe, N. C.

    1986-01-01

    The fossil record of the microscopic algae classified as diatoms suggests they were injected to earth at the Cretaceous boundary. Not only could diatoms remain viable in the cometary environment, but also many species might replicate in illuminated surface layers or early interior layers of cometary ice. Presumably they reached the solar system on an interstellar comet as an already-evolved assemblage of organisms. Diatoms might cause color changes to comet nuclei while their outgassing decays and revives around highly elliptical orbits. Just as for interstellar absorption, high-resolution IR observations are capable of distinguishing whether the 10-micron feature arises from siliceous diatom material or mineral silicates. The 10-30-micron band and the UV 220-nm region can also provide evidence of biological material.

  1. Comet Radar Explorer

    Science.gov (United States)

    Asphaug, Erik; CORE Science Team

    2010-10-01

    Comet Radar Explorer (CORE) is a low cost mission that uses sounding radar to image the 3D internal structure of the nucleus of Jupiter-family comet (JFC) Tempel 2. Believed to originate in the Kuiper Belt, JFCs are among the most primitive bodies in the inner solar system. CORE operates a 5 and 15 MHz Radar Reflection Imager from close orbit about the nucleus of Tempel 2, obtaining a dense network of echoes that are used to map its interior dielectric contrasts to high resolution (ង m) and resolve the dielectric constants to  m throughout the 16x8x9 km nucleus. The resulting clear images of internal structure and composition reveal how the nucleus was formed and how it has evolved. Radiometric tracking of the spacecraft orbit results in an interior mass distribution that constrains the radar-based models of interior composition. High-resolution visible and infrared color images provide the surface and exterior boundary conditions for interior models and hypotheses. They present the geology and morphology of the nucleus surface at meter-scales, and also the time-evolving activity, structure and composition of the inner coma. By making deep connections from interior to exterior, the data CORE provides will answer fundamental questions about the earliest stages of planetesimal evolution and planet formation, and lay the foundation for a comet nucleus sample return mission. CORE is led by Prof. Erik Asphaug of the University of California, Santa Cruz and is managed by JPL. It benefits from key scientific and payload contributions by ASI and CNES. The international science team has been assembled on the basis of their key involvement in past and ongoing missions to comets, and in Mars radar missions, and for their expertise in radar data analysis.

  2. Comets and their origin the tools to decipher a comet

    CERN Document Server

    Meierhenrich, Uwe

    2014-01-01

    Divided into two parts, the first four chapters of Comets and their Origin refer to comets and their formation in general, describing cometary missions, comet remote observations, astrochemistry, artificial comets, and the chirality phenomenon.The second part covers the cometary Rosetta mission, its launch, journey, scientific objectives, and instrumentations, as well as the landing scenario on a cometary nucleus. Along the way, the author presents general questions concerning the origin of terrestrial water and the molecular beginnings of lifeon Earth, as well as how the instruments used on

  3. Disc electrophoresis and related techniques of polyacrylamide gel electrophoresis

    National Research Council Canada - National Science Library

    Maurer, H. R

    1971-01-01

    ..., enzymes, antingens and radioactively labelled materials, and detailed treatments of micro disc electrophoresis, preparative polyacrylamide gel electrophoresis and many other techniques for special problems...

  4. [Risk factors and impacts on prognosis of ultrasound lung comets in patients undergoing hemodialysis].

    Science.gov (United States)

    Wang, Z L; Liu, J Y; Zhou, C J; Wang, M B; Wang, H Y; Xu, Y

    2017-12-26

    Objective: To investigate the risk factors of ultrasound lung comets and its impact on the survivals of patients undergoing hemodialysis. Methods: One hundred and forty-two patients on hemodialysis (Male 76, female 66) were divided into three groups according to the score of lung comets (mild: ≤14 comets; moderate: 15 to 30 comets; severe: >30 comets). Seventy-two healthy subjects examined by lung ultrasound serve as a control. Pulmonary artery systolic pressure and left ventricular ejection fraction (LVEF) were assessed by Doppler ultrasonography. High-sensitivity C-reactive protein (hsCRP) and tumor necrosis factor (TNF)-α were measured by the automatic analyzer and enzyme-linked immunosorbent assay. Results: With the increasing age of the patients, lung comets scores increased ( P comets score was positively related to multiple clinical variables including New York Heart Association (NYHA) classification ( P =0.023), hsCRP ( P =0.042), TNF-α ( P comets score was independent risk factor for death ( P =0.001). In Kaplan-Meier survival analysis, the risk of all-cause mortality increased in parallel with lung comets score, and patients with lung comets score (>30 comets) were at highest risk of death among all three groups (log-rank test χ(2)=12.73, P =0.001). Conclusion: Lung comets is associated with inflammation, pulmonary artery systolic pressure/volume overload and heart function. Lung comets score represents the alterations of heart function and it may serve as a powerful predictor of all-cause mortality for hemodialysis patients.

  5. Technical manual for COMET

    International Nuclear Information System (INIS)

    Song, Jin Ho; Kwon, Young Min; Kim, Taek Mo; Lee, Sang Jong; Jeong, Hae Yong

    1996-07-01

    The purpose of this report is to provide a description for a COMET computer code which is to be used in the analysis of mass and energy releases during post-blowdown phase of LOCA. The mass and energy data re to be used as input data for the containment functional design. This report contains a brief description of analytical models and guidelines for the usage of the computer code. This computer code is to be used for both cold leg and hot leg break analyses. A verification analyses are performed for Ulchin 3 and 4 cold and hot leg break. 11 figs (Author)

  6. Physics of comets

    International Nuclear Information System (INIS)

    Swamy, K.S.K.

    1986-01-01

    This book deals systematically with the physics of comets in the light of our present knowledge about various aspects of cometary phenomena and the problems that require further attention. The basic aspects, methods and models that have been used extensively at the present time for the interpretation of cometary observations are fully discussed. Contents: General Introduction; Orbital Dynamics; Atomic Physics; Spectra; Spectra of Coma; Gas Production Rates; Dust Tail; Light Scattering Theory; Nature of the Dust Particle; Ion Tails; Nucleus; Origins; Problems and Prospects

  7. Inside look at Halley's comet

    International Nuclear Information System (INIS)

    Beatty, J.K.

    1986-01-01

    The 1985-1986 emergence of Halley's comet, the first since the advent of the space age, was explored by a variety of spacecraft. The Vega 1, launched by the USSR together with the Eastern-block alliance, passed 5523 miles from the comet's nucleus at 7:20:06 Universal time. It indicated that the comet was about 300 miles closer to the sun than had been predicted. The Japanese spacecraft, Suisei, was created to map the distribution of neutral hydrogen atoms outside Halley's visible coma. Its pictures indicated that the comet's output of water varied between 25 and 60 tons per second. Five days after the Vega 2's passage through the comet, the Giotto (sponsored by the European Space Agency) probe appeared. Giotto's close approach took place 3.1 minutes after midnight UT on March 14th; the craft had passed 376 miles from its target. Giotto's data indicated that the nucleus was bigger than expected, and that the comet was composed primarily of water, CO2 and N2. The Vegas and Giotto found that as the solar wind approaches Halley, it slows gradually and the solar magnetic lines embedded in the wind begin to pile up. Pick-up ions, from the comet's halo of neutral hydrogen, were found in this solar wind. Sensors on the Vega spacecraft found a variety of plasma waves propagating inside the bow wave. In order to synthesize all the results, a conference on the exploration of Halley's comet will be held this October

  8. Detecting active comets with SDSS

    Energy Technology Data Exchange (ETDEWEB)

    Solontoi, Michael; Ivezic, Zeljko; /Washington U., Seattle, Astron. Dept.; West, Andrew A.; /MIT, MKI; Claire, Mark; /Washington U., Seattle, Astron. Dept.; Juric, Mario; /Princeton U. Observ.; Becker, Andrew; Jones, Lynne; /Washington U., Seattle, Astron. Dept.; Hall, Patrick B.; /York U., Canada; Kent, Steve; /Fermilab; Lupton, Robert H.; /Princeton U. Observ.; Quinn, Tom; /Washington U., Seattle, Astron. Dept. /Princeton U. Observ.

    2010-12-01

    Using a sample of serendipitously discovered active comets in the Sloan Digital Sky Survey (SDSS), we develop well-controlled selection criteria for greatly increasing the efficiency of comet identification in the SDSS catalogs. After follow-up visual inspection of images to reject remaining false positives, the total sample of SDSS comets presented here contains 19 objects, roughly one comet per 10 million other SDSS objects. The good understanding of selection effects allows a study of the population statistics, and we estimate the apparent magnitude distribution to r {approx} 18, the ecliptic latitude distribution, and the comet distribution in SDSS color space. The most surprising results are the extremely narrow range of colors for comets in our sample (e.g. root-mean-square scatter of only {approx}0.06 mag for the g-r color), and the similarity of comet colors to those of jovian Trojans. We discuss the relevance of our results for upcoming deep multi-epoch optical surveys such as the Dark Energy Survey, Pan-STARRS, and the Large Synoptic Survey Telescope (LSST), and estimate that LSST may produce a sample of about 10,000 comets over its 10-year lifetime.

  9. Where Do Comets Come From?

    Science.gov (United States)

    Van Flandern, Tom

    1982-01-01

    Proposes a new origin for comets in the solar system, namely, that comets originated in the breakup of a body orbiting the sun in or near the present location of the asteroid belt in the relatively recent past. Predictions related to the theory are discussed. (Author/JN)

  10. Comet or Asteroid?

    Science.gov (United States)

    1997-11-01

    When is a minor object in the solar system a comet? And when is it an asteroid? Until recently, there was little doubt. Any object that was found to display a tail or appeared diffuse was a comet of ice and dust grains, and any that didn't, was an asteroid of solid rock. Moreover, comets normally move in rather elongated orbits, while most asteroids follow near-circular orbits close to the main plane of the solar system in which the major planets move. However, astronomers have recently discovered some `intermediate' objects which seem to possess properties that are typical for both categories. For instance, a strange object (P/1996 N2 - Elst-Pizarro) was found last year at ESO ( ESO Press Photo 36/96 ) which showed a cometary tail, while moving in a typical asteroidal orbit. At about the same time, American scientists found another (1996 PW) that moved in a very elongated comet-type orbit but was completely devoid of a tail. Now, a group of European scientists, by means of observations carried out at the ESO La Silla observatory, have found yet another object that at first appeared to be one more comet/asteroid example. However, continued and more detailed observations aimed at revealing its true nature have shown that it is most probably a comet . Consequently, it has received the provisional cometary designation P/1997 T3 . The Uppsala-DLR Trojan Survey Some time ago, Claes-Ingvar Lagerkvist (Astronomical Observatory, Uppsala, Sweden), in collaboration with Gerhard Hahn, Stefano Mottola, Magnus Lundström and Uri Carsenty (DLR, Institute of Planetary Exploration, Berlin, Germany), started to study the distribution of asteroids near Jupiter. They were particularly interested in those that move in orbits similar to that of Jupiter and which are located `ahead' of Jupiter in the so-called `Jovian L4 Lagrangian point'. Together with those `behind' Jupiter, these asteroids have been given the names of Greek and Trojan Heroes who participated in the famous Trojan war

  11. Multiwavelength Observations of Recent Comets

    Science.gov (United States)

    Milam, Stefanie N.; Charnley, Steven B.; Gicquel, Adeline; Cordiner, Martin; Kuan, Yi-Jehng; Chuang, Yo-Ling; Villanueva, Geronimo; DiSanti, Michael A.; Bonev, Boncho P.; Remijan, Anthony J.; hide

    2013-01-01

    Comets provide important clues to the physical and chemical processes that occurred during the formation and early evolution of the Solar System, and could also have been important for initiating prebiotic chemistry on the early Earth. Comets are comprised of molecular ices, that may be pristine inter-stellar remnants of Solar System formation, along with high-temperature crystalline silicate dust that is indicative of a more thermally varied history in the protosolar nebula. Comparing abundances of cometary parent volatiles, and isotopic fractionation ratios, to those found in the interstellar medium, in disks around young stars, and between cometary families, is vital to understanding planetary system formation and the processing history experienced by organic matter in the so-called interstellar-comet connection. We will present a comparison of molecular abundances in these comets to those observed in others, supporting a long-term effort of building a comet taxonomy based on composition.

  12. Asteroids, Comets, Meteors 2014

    Science.gov (United States)

    Muinonen, K.; Penttilä, A.; Granvik, M.; Virkki, A.; Fedorets, G.; Wilkman, O.; Kohout, T.

    2014-08-01

    Asteroids, Comets, Meteors focuses on the research of small Solar System bodies. Small bodies are the key to understanding the formation and evolution of the Solar System, carrying signals from pre-solar times. Understanding the evolution of the Solar System helps unveil the evolution of extrasolar planetary systems. Societally, small bodies will be important future resources of minerals. The near-Earth population of small bodies continues to pose an impact hazard, whether it be small pieces of falling meteorites or larger asteroids or cometary nuclei capable of causing global environmental effects. The conference series entitled ''Asteroids, Comets, Meteors'' constitutes the leading international series in the field of small Solar System bodies. The first three conferences took place in Uppsala, Sweden in 1983, 1985, and 1989. The conference is now returning to Nordic countries after a quarter of a century. After the Uppsala conferences, the conference has taken place in Flagstaff, Arizona, U.S.A. in 1991, Belgirate, Italy in 1993, Paris, France in 1996, Ithaca, New York, U.S.A. in 1999, in Berlin, Germany in 2002, in Rio de Janeiro, Brazil in 2005, in Baltimore, Maryland, U.S.A. in 2008, and in Niigata, Japan in 2012. ACM in Helsinki, Finland in 2014 will be the 12th conference in the series.

  13. Evaluation of genotoxic effect of prozac (fluoxetine without and with addition of vitamins A and C by means of the comet assay in culture of CHO-K1 cells

    Directory of Open Access Journals (Sweden)

    Noélle Giacomini Lemos

    2005-12-01

    Full Text Available The fluoxetine, commercially named Prozac, is efficient against depression and anxiety, with lower risk of collateral effects. However, the possible genotoxic effects are still unknown. The use of vitamins as protectors against damages on cells and DNA has been evaluated, mainly for vitamins A and C. Furthermore, the associative effect of vitamins with several medicines demands studies. The evaluations of genotoxic effect of Prozac and vitamins A and C protective effect were carried out in culture of Chinese hamster ovary cells, CHO-K1, by means of the comet test. The Prozac was used, in liquid formulation, diluted in 5µg, 1µg and 0.2 µg/mL of culture medium. The vitamins were used, in liquid formulation, at the concentrations of 3µg and 880,5 µg/mL of culture medium to vitamins A and C, respectively. The treatments were carried out during 1 hour. The obtained data demonstrated that only the highest concentration of Prozac (5 µg is genotoxic and both vitamins A and C reduced such genotoxicity. The data suggest a follow-up on patients who use Prozac and the possibility of vitamins A and C association in order to minimize the collateral genotoxic effects.

  14. Protein electrophoresis - serum

    Science.gov (United States)

    ... this page: //medlineplus.gov/ency/article/003540.htm Protein electrophoresis - serum To use the sharing features on ... JavaScript. This lab test measures the types of protein in the fluid (serum) part of a blood ...

  15. Evaluating the potential genotoxicity of phthalates esters (PAEs) in perfumes using in vitro assays.

    Science.gov (United States)

    Al-Saleh, Iman; Al-Rajudi, Tahreer; Al-Qudaihi, Ghofran; Manogaran, Pulicat

    2017-10-01

    We previously reported high levels of phthalate esters (PAEs) added as solvents or fixatives in 47 brands of perfumes. Diethyl phthalate was the most abundant compound (0.232-23,649 ppm), and 83.3% of the perfumes had levels >1 ppm, the threshold limit cited by a Greenpeace investigation. All samples had dimethyl phthalate levels higher than its threshold limit of 0.1 ppm, and 88, 38, and 7% of the perfumes had benzyl butyl phthalate, di(2-ethylhexyl) phthalate, and dibutyl phthalate levels, respectively, above their threshold limits. The role of PAEs as endocrine disruptors has been well documented, but their effect on genotoxic behavior has received little attention. We used in vitro single-cell gel electrophoresis (comet) and micronucleus (MN) assays with human lymphoblastoid TK6 cells to evaluate the genotoxic potency of 42 of the same perfumes and to determine its association with PAEs. All perfumes induced more DNA damage than a negative control (NEG), ≥ 90% of the samples caused more damage than cells treated with the vehicles possibly used in perfume's preparations such as methanol (ME) and ethanol (ET), and 11.6% of the perfumes caused more DNA damage than a positive control (hydrogen peroxide). Chromosome breakage expressed as MN frequency was higher in cells treated with 71.4, 64.3, 57.1, and 4.8% of the perfumes than in NEG, cells treated with ME or ET, and another positive control (x-rays), respectively. The genotoxic responses in the comet and MN assays were not correlated. The comet assay indicated that the damage in TK6 cells treated with five PAEs at concentrations of 0.05 and 0.2 ppm either individually or as a mixture did not differ significantly from the damage in cells treated with the perfumes. Unlike the comet assay, the sensitivity of the MN assay to PAEs was weak at both low and high concentrations, and MN frequencies were generally low. This study demonstrates for the first time the possible contribution of PAEs in perfumes to DNA

  16. Determination of the Mutagenicity Potential of Supermint Herbal Medicine by Single Cell Gel Electrophoresis in Rat Hepatocytes

    Directory of Open Access Journals (Sweden)

    Zivar Amanpour

    2012-08-01

    Full Text Available Purpose: The increasing use of herbal drugs and their easy availability have necessitated the use of mutagenicity test to analyze their toxicity and safety. The aim of this study was to evaluate the genotoxicity of Supermint herbal medicine in DNA breakage of rat hepatocytes in comparison with sodium dichromate by single cell gel electrophoresis technique or comet assay. Methods: Hepatocytes were prepared from male wistar rats and were counted and kept in a bioreactor for 30 minutes. Then cells were exposed to the Supermint herbal medicine at doses of 125, 250 and 500 μl/ml. Buffer 4 (incubation buffer and sodium dichromate were used as negative and positive control for one hour respectively. Then cell suspension with low melting point agarose were put on precoated slides and covered with agarose gel. Then lysing, electrophoresis, neutralization and staining were carried out. Finally the slides were analyzed with fluorescence microscope. The parameter under this analysis was the type of migration which was determined according to Kobayashi pattern. Results: With increased dose of Supermint herbal medicine the DNA damage was slightly increased (P<0001. Conlusion: In overall compared to the positive control significant differences is observed which convinced that the crude extract of Supermint in vitro did not have mutagenic effect.

  17. EPOXI at comet Hartley 2.

    Science.gov (United States)

    A'Hearn, Michael F; Belton, Michael J S; Delamere, W Alan; Feaga, Lori M; Hampton, Donald; Kissel, Jochen; Klaasen, Kenneth P; McFadden, Lucy A; Meech, Karen J; Melosh, H Jay; Schultz, Peter H; Sunshine, Jessica M; Thomas, Peter C; Veverka, Joseph; Wellnitz, Dennis D; Yeomans, Donald K; Besse, Sebastien; Bodewits, Dennis; Bowling, Timothy J; Carcich, Brian T; Collins, Steven M; Farnham, Tony L; Groussin, Olivier; Hermalyn, Brendan; Kelley, Michael S; Kelley, Michael S; Li, Jian-Yang; Lindler, Don J; Lisse, Carey M; McLaughlin, Stephanie A; Merlin, Frédéric; Protopapa, Silvia; Richardson, James E; Williams, Jade L

    2011-06-17

    Understanding how comets work--what drives their activity--is crucial to the use of comets in studying the early solar system. EPOXI (Extrasolar Planet Observation and Deep Impact Extended Investigation) flew past comet 103P/Hartley 2, one with an unusually small but very active nucleus, taking both images and spectra. Unlike large, relatively inactive nuclei, this nucleus is outgassing primarily because of CO(2), which drags chunks of ice out of the nucleus. It also shows substantial differences in the relative abundance of volatiles from various parts of the nucleus.

  18. COnsortium of METabolomics Studies (COMETS)

    Science.gov (United States)

    The COnsortium of METabolomics Studies (COMETS) is an extramural-intramural partnership that promotes collaboration among prospective cohort studies that follow participants for a range of outcomes and perform metabolomic profiling of individuals.

  19. Lectin affinity electrophoresis.

    Science.gov (United States)

    Kobayashi, Yuka

    2014-01-01

    An interaction or a binding event typically changes the electrophoretic properties of a molecule. Affinity electrophoresis methods detect changes in the electrophoretic pattern of molecules (mainly macromolecules) that occur as a result of biospecific interactions or complex formation. Lectin affinity electrophoresis is a very effective method for the detection and analysis of trace amounts of glycobiological substances. It is particularly useful for isolating and separating the glycoisomers of target molecules. Here, we describe a sensitive technique for the detection of glycoproteins separated by agarose gel-lectin affinity electrophoresis that uses antibody-affinity blotting. The technique is tested using α-fetoprotein with lectin (Lens culinaris agglutinin and Phaseolus vulgaris agglutinin)-agarose gels.

  20. Capillary electrophoresis of diuretics.

    Science.gov (United States)

    Riekkola, M L; Jumppanen, J H

    1996-05-31

    The review surveys the application of capillary electrophoresis to the screening, identification and determination of diuretics and probenecid. The number of publications is still limited, but the studies already published clearly show that capillary zone electrophoresis (CZE) and micellar electrokinetic chromatography are excellent alternatives for the investigation of diuretics. High accuracy identifications of diuretics and probenecid, even in urine samples, can be obtained when CZE is used with the marker techniques. This review paper has been written from the viewpoint of practical use and some hints are given for future CE studies on diuretics.

  1. Detection of telomerase activity using microchip electrophoresis.

    Science.gov (United States)

    Karasawa, Koji; Arakawa, Hidetoshi

    2015-07-01

    Telomerase participates in malignant transformation or immortalization of cells and thus has attracted attention as an anticancer drug target and diagnostic tumor marker. The telomeric repeat amplification protocol (TRAP) and improved TRAP methods (TRAP-fluorescence, TRAP-hybridization, etc.) are widely used forms of this telomerase assay. However, these approaches generally employ acrylamide gel electrophoresis after amplification of telomeric repeats by polymerase chain reaction (PCR), making these TRAP methods time consuming and technically demanding. In this study we developed a novel telomerase assay using microchip electrophoresis for rapid and highly sensitive detection of telomerase activity in cancer cells. The mixed gel of 0.8% hydroxypropyl methylcellulose (HPMC) and 0.3% polyethylene oxide (PEO) with SYBR Gold (fluorescent reagent) was used for microchip electrophoresis. As a result, the product amplified by a telomerase-positive cell could be measured in one cell per assay and detected with high reproducibility (CV=0.67%) in the short time of 100s. Copyright © 2015 Elsevier B.V. All rights reserved.

  2. DNA ELECTROPHORESIS AT SURFACES

    Energy Technology Data Exchange (ETDEWEB)

    RAFAILOVICH, MIRIAM; SOKOLOV, JONATHAN; GERSAPPE, DILIP

    2003-09-01

    During this year we performed two major projects: I. We developed a detailed theoretical model which complements our experiments on surface DNA electrophoresis. We found that it was possible to enhance the separation of DNA chains by imposing a chemical nanoscale pattern on the surface. This approach utilized the surface interaction effect of the DNA chains with the substrate and is a refinement to our previous method in which DNA chains were separated on homogeneous flat surfaces. By introducing the nano-patterns on the surface, the conformational changes of DNA chains of different lengths can be amplified, which results in the different friction strengths with the substrate surface. Our results also show that, when compared to the DNA electrophoresis performed on homogeneous flat surfaces, nanopatterned surfaces offer a larger window in choosing different surface interactions to achieve separation. II. In collaboration with a large international manufacturer of skin care products we also embarked on a project involving photo toxicity of titanium dioxide nanoparticles, which are a key ingredient in sunscreen and cosmetic lotions. The results clearly implicated the nanoparticles in catalyzing damage to chromosomal DNA. We then used this knowledge to develop a polymer/anti-oxidant coating which prevented the photocatalytic reaction on DNA while still retaining the UV absorptive properties of the nanoparticles. The standard gel electrophoresis was not sufficient in determining the extent of the DNA damage. The conclusions of this study were based predominantly on analysis obtained with the surface electrophoresis method.

  3. Tabulation of comet observations.

    Science.gov (United States)

    1993-04-01

    Concerning comets: 1980 IV Černis-Petrauskas, 1985 XVII Hartley-Good, 1985 XIII P/Giacobini-Zinner, 1985 XIX Thiele, 1986 III P/Halley, 1988 I Ichimura, 1988 V Liller, 1988 XV Machholz, 1989 X P/Brorsen-Metcalf, 1989 XIX Okazaki-Levy-Rudenko, 1989 XXII Aarseth-Brewington, 1990 V Austin, 1990 XVII Tsuchiya-Kiuchi, 1990 XX Levy, 1991 I P/Metcalf-Brewington, 1991 XXIV Shoemaker-Levy, 1991l Helin-Lawrence, 1991a1 Shoemaker-Levy, 1991g1 Zanotta-Brewington, 1991h1 Mueller, 1992d Tanaka-Machholz, 1992h Spacewatch, 1992l P/Giclas, 1992m P/Wolf, 1992n P/Faye, 1992p P/Brewington, 1992q Helin-Lawrence, 1992s P/Ciffréo, 1992t P/Swift-Tuttle, 1992u P/Väisälä, 1992x P/Schaumasse, 1992y Shoemaker, 1992a1 Ohshita, 1993a Mueller, 1993d Mueller, 1993e P/Shoemaker-Levy 9, P/Encke, P/Schwassmann-Wachmann 1.

  4. The mass disruption of Oort cloud comets.

    Science.gov (United States)

    Levison, Harold F; Morbidelli, Alessandro; Dones, Luke; Jedicke, Robert; Wiegert, Paul A; Bottke, William F

    2002-06-21

    We have calculated the number of dormant, nearly isotropic Oort cloud comets in the solar system by (i) combining orbital distribution models with statistical models of dormant comet discoveries by well-defined surveys and (ii) comparing the model results to observations of a population of dormant comets. Dynamical models that assume that comets are not destroyed predict that we should have discovered approximately 100 times more dormant nearly isotropic comets than are actually seen. Thus, as comets evolve inward from the Oort cloud, the majority of them must physically disrupt.

  5. Genotoxicity of indium tin oxide by comet test

    Directory of Open Access Journals (Sweden)

    İbrahim Hakkı Ciğerci

    2015-06-01

    Full Text Available Indium tin oxide (ITO is used for liquid crystal display (LCDs, electrochromic displays, flat panel displays, field emission displays, touch or laptop computer screens, cell phones, energy conserving architectural windows, defogging aircraft and automobile windows, heat-reflecting coatings to increase light bulb efficiency, gas sensors, antistatic window coatings, wear resistant layers on glass, nanowires and nanorods because of its unique properties of high electrical conductivity, transparency and mechanical resistance.Genotoxic effects of ITO were investigated on the root cells of Allium cepa by Comet assay. A. cepa roots were treated with the aqueous dispersions of ITO at 5 different concentrations (12.5, 25, 50, 75, and 100 ppm for 4 h. A significant increase in DNA damage was a observed at all concentrations of ITO by Comet assay. These result indicate that ITO exhibit genotoxic activity in A. cepa root meristematic cells.

  6. An Evaluation of Quantitative PCR Assays (TaqMan® and SYBR Green for the Detection of Babesia bigemina and Babesia bovis, and a Novel Fluorescent-ITS1-PCR Capillary Electrophoresis Method for Genotyping B. bovis Isolates

    Directory of Open Access Journals (Sweden)

    Bing Zhang

    2016-09-01

    Full Text Available Babesia spp. are tick-transmitted haemoparasites causing tick fever in cattle. In Australia, economic losses to the cattle industry from tick fever are estimated at AUD$26 Million per annum. If animals recover from these infections, they become immune carriers. Here we describe a novel multiplex TaqMan qPCR targeting cytochrome b genes for the identification of Babesia spp. The assay shows high sensitivity, specificity and reproducibility, and allows quantification of parasite DNA from Babesia bovis and B. bigemina compared to standard PCR assays. A previously published cytochrome b SYBR Green qPCR was also tested in this study, showing slightly higher sensitivity than the Taqman qPCRs but requires melting curve analysis post-PCR to confirm specificity. The SYBR Green assays were further evaluated using both diagnostic submissions and vaccinated cattle (at 7, 9, 11 and 14 days post-inoculation showed that B. bigemina can be detected more frequently than B. bovis. Due to fewer circulating parasites, B. bovis detection in carrier animals requires higher DNA input. Preliminary data for a novel fluorescent PCR genotyping based on the Internal Transcribed Spacer 1 region to detect vaccine and field alleles of B. bovis are described. This assay is capable of detecting vaccine and novel field isolate alleles in a single sample.

  7. Comet showers and Nemesis, the death star

    International Nuclear Information System (INIS)

    Hills, J.G.

    1984-01-01

    The recently proposed hypothesis that the periodic extinctions of terrestrial species are the result of comet showers catalyzed by a hypothetical distant solar companion, Nemesis, a tale of global death by comet bombardment of the earth, is discussed

  8. Practical capillary electrophoresis

    CERN Document Server

    Weinberger, Robert

    2000-01-01

    In the 1980s, capillary electrophoresis (CE) joined high-performance liquid chromatography (HPLC) as the most powerful separation technique available to analytical chemists and biochemists. Published research using CE grew from 48 papers in the year of commercial introduction (1988) to 1200 in 1997. While only a dozen major pharmaceutical and biotech companies have reduced CE to routine practice, the applications market is showing real or potential growth in key areas, particularly in the DNA marketplace for genomic mapping and forensic identification. For drug development involving small molecules (including chiral separations), one CE instrument can replace 10 liquid chromatographs in terms of speed of analysis. CE also uses aqueous rather than organic solvents and is thus environmentally friendlier than HPLC. The second edition of Practical Capillary Electrophoresis has been extensively reorganized and rewritten to reflect modern usage in the field, with an emphasis on commercially available apparatus and ...

  9. Analysis of electrophoresis performance

    Science.gov (United States)

    Roberts, G. O.

    1984-01-01

    The SAMPLE computer code models electrophoresis separation in a wide range of conditions. Results are included for steady three dimensional continuous flow electrophoresis (CFE), time dependent gel and acetate film experiments in one or two dimensions and isoelectric focusing in one dimension. The code evolves N two dimensional radical concentration distributions in time, or distance down a CFE chamber. For each time or distance increment, there are six stages, successively obtaining the pH distribution, the corresponding degrees of ionization for each radical, the conductivity, the electric field and current distribution, and the flux components in each direction for each separate radical. The final stage is to update the radical concentrations. The model formulation for ion motion in an electric field ignores activity effects, and is valid only for low concentrations; for larger concentrations the conductivity is, therefore, also invalid.

  10. CATASTROPHIC DISRUPTION OF COMET ISON

    Energy Technology Data Exchange (ETDEWEB)

    Keane, Jacqueline V.; Kleyna, Jan T.; Riesen, Timm-Emmanuel; Meech, Karen J. [Institute for Astronomy, University of Hawaii, 2680 Woodlawn Drive, Honolulu, HI 96822 (United States); Milam, Stefanie N.; Charnley, Steven B. [Astrochemistry Laboratory, NASA GSFC, MS 690, Greenbelt, MD 20771 (United States); Coulson, Iain M. [Joint Astronomy Center, 660 North Aohoku Place, Hilo, HI 96720 (United States); Sekanina, Zdenek [Jet Propulsion Laboratory, California Institute of Technology, 4800 Oak Grove Drive, Pasadena, CA 91109 (United States); Kracht, Rainer, E-mail: keane@ifa.hawaii.edu [Ostlandring 53, D-25335 Elmshorn, Schleswig-Holstein (Germany)

    2016-11-10

    We report submillimeter 450 and 850 μ m dust continuum observations for comet C/2012 S1 (ISON) obtained at heliocentric distances 0.31–0.08 au prior to perihelion on 2013 November 28 ( r {sub h} = 0.0125 au). These observations reveal a rapidly varying dust environment in which the dust emission was initially point-like. As ISON approached perihelion, the continuum emission became an elongated dust column spread out over as much as 60″ (>10{sup 5} km) in the anti-solar direction. Deconvolution of the November 28.04 850 μ m image reveals numerous distinct clumps consistent with the catastrophic disruption of comet ISON, producing ∼5.2 × 10{sup 10} kg of submillimeter-sized dust. Orbital computations suggest that the SCUBA-2 emission peak coincides with the comet's residual nucleus.

  11. Catastrophic Disruption of Comet ISON

    Science.gov (United States)

    Keane, Jacqueline V.; Milam, Stefanie N.; Coulson, Iain M.; Kleyna, Jan T.; Sekanina, Zdenek; Kracht, Rainer; Riesen, Timm-Emmanuel; Meech, Karen J.; Charnley, Steven B.

    2016-01-01

    We report submillimeter 450 and 850 microns dust continuum observations for comet C/2012 S1 (ISON) obtained at heliocentric distances 0.31-0.08 au prior to perihelion on 2013 November 28 (rh?=?0.0125 au). These observations reveal a rapidly varying dust environment in which the dust emission was initially point-like. As ISON approached perihelion, the continuum emission became an elongated dust column spread out over as much as 60? (greater than 10(exp 5) km in the anti-solar direction. Deconvolution of the November 28.04 850 microns image reveals numerous distinct clumps consistent with the catastrophic disruption of comet ISON, producing approximately 5.2?×?10(exp 10) kg of submillimeter-sized dust. Orbital computations suggest that the SCUBA-2 emission peak coincides with the comet's residual nucleus.

  12. Hummingbird Comet Nucleus Analysis Mission

    Science.gov (United States)

    Kojiro, Daniel; Carle, Glenn C.; Lasher, Larry E.

    2000-01-01

    Hummingbird is a highly focused scientific mission, proposed to NASA s Discovery Program, designed to address the highest priority questions in cometary science-that of the chemical composition of the cometary nucleus. After rendezvous with the comet, Hummingbird would first methodically image and map the comet, then collect and analyze dust, ice and gases from the cometary atmosphere to enrich characterization of the comet and support landing site selection. Then, like its namesake, Hummingbird would carefully descend to a pre-selected surface site obtaining a high-resolution image, gather a surface material sample, acquire surface temperature and then immediately return to orbit for detailed chemical and elemental analyses followed by a high resolution post-sampling image of the site. Hummingbird s analytical laboratory contains instrumentation for a comprehensive molecular and elemental analysis of the cometary nucleus as well as an innovative surface sample acquisition device.

  13. CO2 Orbital Trends in Comets

    Science.gov (United States)

    Kelley, Michael; Feaga, Lori; Bodewits, Dennis; McKay, Adam; Snodgrass, Colin; Wooden, Diane

    2014-12-01

    Spacecraft missions to comets return a treasure trove of details of their targets, e.g., the Rosetta mission to comet 67P/Churyumov-Gerasimenko, the Deep Impact experiment at comet 9P/Tempel 1, or even the flyby of C/2013 A1 (Siding Spring) at Mars. Yet, missions are rare, the diversity of comets is large, few comets are easily accessible, and comet flybys essentially return snapshots of their target nuclei. Thus, telescopic observations are necessary to place the mission data within the context of each comet's long-term behavior, and to further connect mission results to the comet population as a whole. We propose a large Cycle 11 project to study the long-term activity of past and potential future mission targets, and select bright Oort cloud comets to infer comet nucleus properties, which would otherwise require flyby missions. In the classical comet model, cometary mass loss is driven by the sublimation of water ice. However, recent discoveries suggest that the more volatile CO and CO2 ices are the likely drivers of some comet active regions. Surprisingly, CO2 drove most of the activity of comet Hartley 2 at only 1 AU from the Sun where vigorous water ice sublimation would be expected to dominate. Currently, little is known about the role of CO2 in comet activity because telluric absorptions prohibit monitoring from the ground. In our Cycle 11 project, we will study the CO2 activity of our targets through IRAC photometry. In conjunction with prior observations of CO2 and CO, as well as future data sets (JWST) and ongoing Earth-based projects led by members of our team, we will investigate both long-term activity trends in our target comets, with a particular goal to ascertain the connections between each comet's coma and nucleus.

  14. Infrared techniques for comet observations

    Science.gov (United States)

    Hanner, Martha S.; Tokunaga, Alan T.

    1991-01-01

    The infrared spectral region (1-1000 microns) is important for studies of both molecules and solid grains in comets. Infrared astronomy is in the midst of a technological revolution, with the development of sensitive 2D arrays leading to IR cameras and spectrometers with vastly improved sensitivity and resolution. The Halley campaign gave us tantalizing first glimpses of the comet science possible with this new technology, evidenced, for example, by the many new spectral features detected in the infrared. The techniques of photometry, imaging, and spectroscopy are reviewed in this chapter and their status at the time of the Halley observations is described.

  15. Species Identification and Strain Typing of Staphylococcus agnetis and Staphylococcus hyicus Isolates from Bovine Milk by Use of a Novel Multiplex PCR Assay and Pulsed-Field Gel Electrophoresis.

    Science.gov (United States)

    Adkins, P R F; Middleton, J R; Calcutt, M J; Stewart, G C; Fox, L K

    2017-06-01

    Staphylococcus hyicus and Staphylococcus agnetis are two coagulase-variable staphylococcal species that can be isolated from bovine milk and are difficult to differentiate. The objectives of this study were to characterize isolates of bovine milk origin from a collection that had previously been characterized as coagulase-positive S. hyicus based on phenotypic species identification methods and to develop a PCR-based method for differentiating S. hyicus , S. agnetis , and Staphylococcus aureus Isolates ( n = 62) were selected from a previous study in which milk samples were collected from cows on 15 dairy herds. Isolates were coagulase tested and identified to the species level using housekeeping gene sequencing. A multiplex PCR to differentiate S. hyicus , S. agnetis , and S. aureus was developed. Pulsed-field gel electrophoresis was conducted to strain type the isolates. Based on gene sequencing, 44/62 of the isolates were determined to be either S. agnetis ( n = 43) or S. hyicus ( n = 1). Overall, 88% (37/42) of coagulase-positive S. agnetis isolates were found to be coagulase positive at 4 h. The herd-level prevalence of coagulase-positive S. agnetis ranged from 0 to 2.17%. Strain typing identified 23 different strains. Six strains were identified more than once and from multiple cows within the herd. Three strains were isolated from cows at more than one time point, with 41 to 264 days between samplings. These data suggest that S. agnetis is likely more prevalent on dairy farms than S. hyicus Also, some S. agnetis isolates in this study appeared to be contagious and associated with persistent infections. Copyright © 2017 American Society for Microbiology.

  16. Comet Mineralogy as Inferred from Infrared Spectra of Comets

    Science.gov (United States)

    Wooden, Diane H.

    2006-01-01

    For most comets, infrared (IR) spectroscopy (remote sensing) is the method through which we diagnose the mineralogy and size distribution of dust in their comae. The shape and contrast of the IR spectral features depend on the particle size: optically active minerals (absorbing of visible and near-IR solar photons) and submicron solid grains or highly porous (> 90% vacuum) grains primarily contribute to the shapes of the observed resonances. Comet mineralogies typically are determined by fitting thermal emission models of ensembles of discrete mineral grains to observed IR spectral energy distributions. The absorptivities (Q-abs) and scattering efficiencies (Q-scat) of the discrete mineral grains are computed using Mie scattering, Maxwell-Garnet mixing, Discrete Dipole Approximation, and Multi-Layered Sphere codes. These techniques when applied to crystalline minerals, specifically olivine (Mg_x, Fe_1-x)2 Si04, x>0.9, require the use of ellipsoidal shaped particles with elongated axial ratios or hollow spheres to produce the shapes of the resonances observed both from comet comae and laboratory samples. The wavelength positions of the distinct resonances from submicron-radii crystalline silicates, as well as their thermal equilibrium temperatures, constrain the crystalline olivine to have a relatively high Mg-content (x>0.9, or Fo>90). Only resonances computed for submicron Mg-rich crystalline olivine and crystalline orthopyroxene match the observed IR spectral features. However, this has led to the interpretation that micron-radii and larger crystals are absent from comet comae. Furthermore, the mass fraction of silicate crystals is dependent upon whether just the submicron portion of the size distribution is being compared or the submicron crystals compare to the aggregates of porous amorphous silicates that are computationally tractable as porous spheres. We will discuss the Deep Impact results as examples of these challenges to interpreting mid-IR spectra of

  17. The COMET Sleep Research Platform.

    Science.gov (United States)

    Nichols, Deborah A; DeSalvo, Steven; Miller, Richard A; Jónsson, Darrell; Griffin, Kara S; Hyde, Pamela R; Walsh, James K; Kushida, Clete A

    2014-01-01

    The Comparative Outcomes Management with Electronic Data Technology (COMET) platform is extensible and designed for facilitating multicenter electronic clinical research. Our research goals were the following: (1) to conduct a comparative effectiveness trial (CET) for two obstructive sleep apnea treatments-positive airway pressure versus oral appliance therapy; and (2) to establish a new electronic network infrastructure that would support this study and other clinical research studies. The COMET platform was created to satisfy the needs of CET with a focus on creating a platform that provides comprehensive toolsets, multisite collaboration, and end-to-end data management. The platform also provides medical researchers the ability to visualize and interpret data using business intelligence (BI) tools. COMET is a research platform that is scalable and extensible, and which, in a future version, can accommodate big data sets and enable efficient and effective research across multiple studies and medical specialties. The COMET platform components were designed for an eventual move to a cloud computing infrastructure that enhances sustainability, overall cost effectiveness, and return on investment.

  18. Comet C/2001 J1

    Czech Academy of Sciences Publication Activity Database

    Pravec, Petr; Helin, E.; Lawrence, K.; Kotková, Lenka; Tichá, J.; Tichý, M.

    č. 7623 (2001), s. 1 ISSN 0081-0304 R&D Projects: GA ČR GA205/99/0255 Institutional research plan: CEZ:AV0Z1003909 Keywords : comets * astrometry Subject RIV: BN - Astronomy, Celestial Mechanics, Astrophysics

  19. Modeling Formaldehyde Emission in Comets

    Science.gov (United States)

    Disanti, M. A.; Reuter, D. C.; Bonev, B. P.; Mumma, M. J.; Villanueva, G. L.

    Modeling fluorescent emission from monomeric formaldehyde (H2CO) forms an integral part of our overall comprehensive program of measuring the volatile composition of comets through high-resolution (RP ~ 25,000) infrared spectroscopy using CSHELL at the IRTF and NIRSPEC at Keck II. The H2CO spectra contain lines from both the nu1 (symmetric CH2 stretch) and nu5 (asymmetric CH2 stretch) bands near 3.6 microns. We have acquired high-quality spectra of twelve Oort cloud comets, and at least six of these show clear emission from H2CO. We also detected H2CO with NIRSPEC in one Jupiter Family comet, 9P/Tempel 1, during Deep Impact observations. Our H2CO model, originally developed to interpret low-resolution spectra of comets Halley and Wilson (Reuter et al. 1989 Ap J 341:1045), predicts individual line intensities (g-factors) as a function of rotational temperature for approximately 1300 lines having energies up to approximately 400 cm^-1 above the ground state. Recently, it was validated through comparison with CSHELL spectra of C/2002 T7 (LINEAR), where newly developed analyses were applied to obtain robust determinations of both the rotational temperature and abundance of H2CO (DiSanti et al. 2006 Ap J 650:470). We are currently in the process of extending the model to higher rotational energy (i.e., higher rotational quantum number) in an attempt to improve the fit to high-J lines in our spectra of C/T7 and other comets. Results will be presented, and implications discussed.Modeling fluorescent emission from monomeric formaldehyde (H2CO) forms an integral part of our overall comprehensive program of measuring the volatile composition of comets through high-resolution (RP ~ 25,000) infrared spectroscopy using CSHELL at the IRTF and NIRSPEC at Keck II. The H2CO spectra contain lines from both the nu1 (symmetric CH2 stretch) and nu5 (asymmetric CH2 stretch) bands near 3.6 microns. We have acquired high-quality spectra of twelve Oort cloud comets, and at least six of

  20. Multiplexed Western Blotting Using Microchip Electrophoresis.

    Science.gov (United States)

    Jin, Shi; Furtaw, Michael D; Chen, Huaxian; Lamb, Don T; Ferguson, Stephen A; Arvin, Natalie E; Dawod, Mohamed; Kennedy, Robert T

    2016-07-05

    Western blotting is a commonly used protein assay that combines the selectivity of electrophoretic separation and immunoassay. The technique is limited by long time, manual operation with mediocre reproducibility, and large sample consumption, typically 10-20 μg per assay. Western blots are also usually used to measure only one protein per assay with an additional housekeeping protein for normalization. Measurement of multiple proteins is possible; however, it requires stripping membranes of antibody and then reprobing with a second antibody. Miniaturized alternatives to Western blot based on microfluidic or capillary electrophoresis have been developed that enable higher-throughput, automation, and greater mass sensitivity. In one approach, proteins are separated by electrophoresis on a microchip that is dragged along a polyvinylidene fluoride membrane so that as proteins exit the chip they are captured on the membrane for immunoassay. In this work, we improve this method to allow multiplexed protein detection. Multiple injections made from the same sample can be deposited in separate tracks so that each is probed with a different antibody. To further enhance multiplexing capability, the electrophoresis channel dimensions were optimized for resolution while keeping separation and blotting times to less than 8 min. Using a 15 μm deep × 50 μm wide × 8.6 cm long channel, it is possible to achieve baseline resolution of proteins that differ by 5% in molecular weight, e.g., ERK1 (44 kDa) from ERK2 (42 kDa). This resolution allows similar proteins detected by cross-reactive antibodies in a single track. We demonstrate detection of 11 proteins from 9 injections from a single Jurkat cell lysate sample consisting of 400 ng of total protein using this procedure. Thus, multiplexed Western blots are possible without cumbersome stripping and reprobing steps.

  1. ISO's analysis of Comet Hale-Bopp

    Science.gov (United States)

    1997-03-01

    The European Space Agency's Infrared Space Observatory ISO inspected Comet Hall-Bopp during the spring and autumn of 1996. The need to keep ISO's telescope extremely cold restricts the spacecraft's pointing in relation to the Sun and the Earth and it ruled out observations at other times. The analyses of the 1996 observations are not yet complete, but already they give new insight into the nature of comets. Comet Hale-Bopp is believed to be a large comet with a nucleus up to 40 kilometres wide. It was discovered in July 1995 by two American astronomers working independently, Alan Hale and Thomas Bopp. At that time, the comet was a billion kilometres away from the Sun, but 200 times brighter than Halley's Comet was, when at a comparable distance. Comet Hale-Bopp will make its closest approach to the Earth on 22 March, and its closest approach to the Sun (perihelion) on 1 April 1997. Some scientific results from ISO The discovery of Comet Hale-Bopp occurred before ISO's launch in November 1995. When first observed by ISO in March and April 1996, the comet was still 700 million kilometres from the Sun, and almost as far from the Earth and ISO. With its privileged view of infrared wavebands inaccessible from the Earth's surface, ISO's photometer ISOPHOT discovered that carbon dioxide was an important constituent of the comet's emissions of vapour.ISOPHOT measured the temperature of the dust cloud around Comet Hale-Bopp. In March 1996, when the comet was still more than 700 million kilometres from the Sun, the dust cloud was at minus 120 degrees C. When ISOPHOT made similar observations in October 1996, the comet was 420 million kilometres from the Sun, and the dust cloud had warmed to about minus 50 degrees C. Intensive observations of Comet Hale-Bopp were also made by ISO's Short-Wave Spectrometer SWS, the Long-Wave Spectrometer LWS, and the ISOPHOT spectrometer PHOT-S. Results are due for publication at the end of March. They will give details about the composition

  2. Comet Halley: nucleus and jets

    International Nuclear Information System (INIS)

    Sagdeev, R.Z.; Avanesov, G.A.; Barinov, I.V.

    1986-06-01

    The VEGA-1 and VEGA-2 spacecrafts made their closest approach to Comet Halley on 6 and 9 March, respectively. In this paper results of the onboard imaging experiment are discussed. The nucleus of the comet was clearly identifyable as an irregularly shaped object with overall dimensions of (16+-1)x(8+-1)x(8+-1) km. The nucleus rotates around its axis which is nearly perpendicular to the orbital plane, with a period of 53+-2 hours. Its albedo is only 0.04+-002. Most of the jet features observed during the second fly-by were spatially reconstructed. These sources form a quasi-linear structure on the surface. The dust above the surface is shown to be optically thin except certain specific dust jets. Brightness features on the surface are clearly seen. Correlating the data with other measurements it is concluded that the dirty snow-ball model probably has to be revised. (author)

  3. Rationalization of Comet Halley's periods

    Science.gov (United States)

    Belton, Michael J. S.

    1990-01-01

    The sense of long axis orientation of Comet Halley during the Vega 1 encounter must be reversed from that deduced by Sagdeev et al. (1986) in order to harmonize the comet nucleus' Vega/Giotto-observed orientations with periodicities extracted from time-series brightness data. It is also demonstrated that Vega/Giotto observations can be satisfied by either a 2.2- or 3.7-day long-axis free precession period. A novel Fourier algorithm is used to reanalyze five independent data sets; strong evidence is adduced for periods harmonically related to a 7.4-day period. The preferred candidate models for Halley's nuclear rotation are characterized by a long-axis precession period of 3.7 days.

  4. Applications of the comet assay in particle toxicology

    DEFF Research Database (Denmark)

    Møller, Peter; Hemmingsen, Jette Gjerke; Jensen, Ditte Marie

    2015-01-01

    Exposure to ambient air particles is associated with elevated levels of DNA strand breaks (SBs) and endonuclease III, formamidopyrimidine DNA glycosylase (FPG) and oxoguanine DNA glycosylase-sensitive sites in cell cultures, animals and humans. In both animals and cell cultures, increases in SB...... to be associated with generation of FPG-sensitive sites in cell cultures, while large differences in SB generation between studies have been noted. Single-dose airway exposure to nanosized carbon black and multi-walled carbon nanotubes in animal models seems to be associated with elevated DNA damage levels in lung...

  5. DNA repair in plants studied by comet assay

    Directory of Open Access Journals (Sweden)

    Karel J Angelis

    2015-06-01

    Fig. 2B. Effect of mutation and of inhibitors of PARP1 on SSB repair kinetics. SSBs induced by 1 hr treatment with 2 mM MMS in atparp1 (red and in Arabidopsis wt in presence of 3 mM 3-aminobenzamide (3-ABA, turquoise and 10 μM HsPARP1 specific AG14361 (green inhibitors. (Angelis and Kozák, unpublished data

  6. Solar wind stagnation near comets

    International Nuclear Information System (INIS)

    Galeev, A.A.; Cravens, T.E.; Gombosi, T.I.

    1983-03-01

    The nature of the solar wind flow near comets is examined analytically. In particular, the typical values for the stagnation pressure and magnetic barrier strength are estimated, taking into account the magnetic field line tension and the charge exchange cooling of the mass loaded solar wind. Knowledge of the strength of the magnetic barrier is required in order to determine the location of the contact discontinuity which separates the contaminated solar wind plasma and the outflowing plasma of the cometary ionosphere. (author)

  7. [Interpopulation defferences in parameters of hemocyte DNA-comets of snail Lymnaea stagnalis from regions with the different environmental load].

    Science.gov (United States)

    Koneva, O Iu

    2013-01-01

    The research of hemocytes of snail Lymnaea stagnalis from regions with different environmental load has been carried out by means of DNA-comet assay. Significant interpopulation distinctions in parameters of hemocytes DNA comets, and also significant differences of sensitivity of hemocyte genetic matherial in snails form different ecological zones to the influence of external damaging factors (in particular, heavy metals) have been revealed by means of the software analysis of hemocyte DNA-comet images. Since the two populations of mollusks are characterized by high genetic identity, the different levels of proliferative processes in hemocytes of snail Lymnaea stagnalis from different ecological zones (that we revealed using the comet assay) may act as an indicator of the intensity of damaging effects and environmental quality.

  8. Comet coma sample return instrument

    Science.gov (United States)

    Albee, A. L.; Brownlee, Don E.; Burnett, Donald S.; Tsou, Peter; Uesugi, K. T.

    1994-01-01

    The sample collection technology and instrument concept for the Sample of Comet Coma Earth Return Mission (SOCCER) are described. The scientific goals of this Flyby Sample Return are to return to coma dust and volatile samples from a known comet source, which will permit accurate elemental and isotopic measurements for thousands of individual solid particles and volatiles, detailed analysis of the dust structure, morphology, and mineralogy of the intact samples, and identification of the biogenic elements or compounds in the solid and volatile samples. Having these intact samples, morphologic, petrographic, and phase structural features can be determined. Information on dust particle size, shape, and density can be ascertained by analyzing penetration holes and tracks in the capture medium. Time and spatial data of dust capture will provide understanding of the flux dynamics of the coma and the jets. Additional information will include the identification of cosmic ray tracks in the cometary grains, which can provide a particle's process history and perhaps even the age of the comet. The measurements will be made with the same equipment used for studying micrometeorites for decades past; hence, the results can be directly compared without extrapolation or modification. The data will provide a powerful and direct technique for comparing the cometary samples with all known types of meteorites and interplanetary dust. This sample collection system will provide the first sample return from a specifically identified primitive body and will allow, for the first time, a direct method of matching meteoritic materials captured on Earth with known parent bodies.

  9. Halley comet, implication on the origin

    International Nuclear Information System (INIS)

    Festou, M.C.

    1990-01-01

    One will first give a rapid description of the different parts that compose a comet coma. Then one will describe the spectrum of comets from the UV to the IR regions with special emphasis on how information relative to the physico-chemistry of comet atmospheres can be retrieved. Our basic knowledge about the composition of comets before 1985 will be summarized and the input of the 1985-86 observing campaign of comet Halley will be shown (in situ, ground-based and space borne observations). One will see then that the chemical composition of comets appears as of today completely compatible with a formation from pre-solar matter that condensed inside the solar system [fr

  10. EVIDENCE FOR COMET STORMS IN METEORITE AGES

    Energy Technology Data Exchange (ETDEWEB)

    Perlmutter, S.; Muller, R.A.

    1987-10-01

    Clustering of cosmic-ray exposure ages of H chondritic meteorites occurs at 7 {+-} 3 and 30 {+-} 6 Myr ago. There is independent evidence that comet storms have occurred at the same times, based on the fossil record of family and genus extinctions, impact craters and glass, and geomagnetic reversals. We suggest that H chondrites were formed by the impact of shower comets on asteroids. The duration of the most recent comet shower was {le} 4 Myr, in agreement with storm theory.

  11. Cometary Volatiles and the Origin of Comets

    Science.gov (United States)

    A'Hearn, Michael F.; Feaga, Lori M.; Keller, H. Uwe; Kawakita, Hideyo; Hampton, Donald L.; Kissel, Jochen; Klaasen, Kenneth P.; McFadden, Lucy A.; Meech, Karen J.; Schultz, Peter H.; hide

    2012-01-01

    We describe recent results on the CO/C02/H2O composition of comets and compare these with models of the protoplanetary disk. We argue that the cometary observations require reactions on grain surfaces to convert CO to CO2 and also require formation between the CO and CO2 snow lines. This then requires very early mixing of cometesimals in the protoplanetary disk analogous to the mixing described for the asteroid belt by Walsh and Morbidelli. We suggest that most comets formed in the region of the giant planets. the traditional source of the Oort-cloud comets but not of the Jupiter-family comets

  12. Advances in capillary electrophoresis and the implications for drug discovery.

    Science.gov (United States)

    Ouimet, Claire M; D'amico, Cara I; Kennedy, Robert T

    2017-02-01

    Many screening platforms are prone to assay interferences that can be avoided by directly measuring the target or enzymatic product. Capillary electrophoresis (CE) and microchip electrophoresis (MCE) have been applied in a variety of formats to drug discovery. CE provides direct detection of the product allowing for the identification of some forms of assay interference. The high efficiency, rapid separations, and low volume requirements make CE amenable to drug discovery. Areas covered: This article describes advances in capillary electrophoresis throughput, sample introduction, and target assays as they pertain to drug discovery and screening. Instrumental advances discussed include integrated droplet microfluidics platforms and multiplexed arrays. Applications of CE to assays of diverse drug discovery targets, including enzymes and affinity interactions are also described. Expert opinion: Current screening with CE does not fully take advantage of the throughputs or low sample volumes possible with CE and is most suitable as a secondary screening method or for screens that are inaccessible with more common platforms. With further development, droplet microfluidics coupled to MCE could take advantage of the low sample requirements by performing assays on the nanoliter scale at high throughput.

  13. Denaturing gradient gel electrophoresis

    International Nuclear Information System (INIS)

    Kocherginskaya, S.A.; Cann, I.K.O.; Mackie, R.I.

    2005-01-01

    It is worthwhile considering that only some 30 species make up the bulk of the bacterial population in human faeces at any one time based on the classical cultivation-based approach. The situation in the rumen is similar. Thus, it is practical to focus on specific groups of interest within the complex community. These may be the predominant or the most active species, specific physiological groups or readily identifiable (genetic) clusters of phylogenetically related organisms. Several 16S rDNA fingerprinting techniques can be invaluable for selecting and monitoring sequences or phylogenetic groups of interest and are described below. Over the past few decades, considerable attention was focussed on the identification of pure cultures of microbes on the basis of genetic polymorphisms of DNA encoding rRNA such as ribotyping, amplified fragment length polymorphism and randomly amplified polymorphic DNA. However, many of these methods require prior cultivation and are less suitable for use in analysis of complex mixed populations although important in describing cultivated microbial diversity in molecular terms. Much less attention was given to molecular characterization of complex communities. In particular, research into diversity and community structure over time has been revolutionized by the advent of molecular fingerprinting techniques for complex communities. Denaturing or temperature gradient gel electrophoresis (DGGE/TGGE) methods have been successfully applied to the analysis of human, pig, cattle, dog and rodent intestinal populations

  14. Comet Dust After Deep Impact

    Science.gov (United States)

    Wooden, Diane H.; Harker, David E.; Woodward, Charles E.

    2006-01-01

    When the Deep Impact Mission hit Jupiter Family comet 9P/Tempel 1, an ejecta crater was formed and an pocket of volatile gases and ices from 10-30 m below the surface was exposed (A Hearn et aI. 2005). This resulted in a gas geyser that persisted for a few hours (Sugita et al, 2005). The gas geyser pushed dust grains into the coma (Sugita et a1. 2005), as well as ice grains (Schulz et al. 2006). The smaller of the dust grains were submicron in radii (0-25.3 micron), and were primarily composed of highly refractory minerals including amorphous (non-graphitic) carbon, and silicate minerals including amorphous (disordered) olivine (Fe,Mg)2SiO4 and pyroxene (Fe,Mg)SiO3 and crystalline Mg-rich olivine. The smaller grains moved faster, as expected from the size-dependent velocity law produced by gas-drag on grains. The mineralogy evolved with time: progressively larger grains persisted in the near nuclear region, having been imparted with slower velocities, and the mineralogies of these larger grains appeared simpler and without crystals. The smaller 0.2-0.3 micron grains reached the coma in about 1.5 hours (1 arc sec = 740 km), were more diverse in mineralogy than the larger grains and contained crystals, and appeared to travel through the coma together. No smaller grains appeared at larger coma distances later (with slower velocities), implying that if grain fragmentation occurred, it happened within the gas acceleration zone. These results of the high spatial resolution spectroscopy (GEMINI+Michelle: Harker et 4. 2005, 2006; Subaru+COMICS: Sugita et al. 2005) revealed that the grains released from the interior were different from the nominally active areas of this comet by their: (a) crystalline content, (b) smaller size, (c) more diverse mineralogy. The temporal changes in the spectra, recorded by GEMIM+Michelle every 7 minutes, indicated that the dust mineralogy is inhomogeneous and, unexpectedly, the portion of the size distribution dominated by smaller grains has

  15. Spectrophotometry of 25 comets - Post-Halley updates for 17 comets plus new observations for eight additional comets

    Science.gov (United States)

    Newburn, Ray L., Jr.; Spinrad, Hyron

    1989-01-01

    The best possible production figures within the current post-Halley framework and available observations are given for H2O, O(1D), CN, C3, C2 and dust in 25 comets. Of these, the three objects with the smallest mixing ratios of all minor species have moderate to little or no dust and appear 'old'. Comets with large amounts of CN are very dusty, and there is a clear correlation of CN with dust, although comets with little or no dust still have some CN. Thus, CN appears to have at least two sources, dust and one or more parent gases. Also, the C2/CN production ratio changes continuously with heliocentric distance in every comet considered, suggesting that C2 production may be a function of coma density as well as parental abundance. Dust production ranges from essentially zero in Comet Sugano-Saigusa-Fujikawa up to 67,000 kg/s for Halley on March 14, 1986.

  16. Amino Acids from a Comet

    Science.gov (United States)

    Cook, Jamie Elisla

    2009-01-01

    NASA's Stardust spacecraft returned samples from comet 81P/Wild 2 to Earth in January 2006. Examinations of the organic compounds in cometary samples can reveal information about the prebiotic organic inventory present on the early Earth and within the early Solar System, which may have contributed to the origin of life. Preliminary studies of Stardust material revealed the presence of a suite of organic compounds including several amines and amino acids, but the origin of these compounds (cometary- vs. terrestrial contamination) could not be identified. We have recently measured the carbon isotopic ratios of these amino acids to determine their origin, leading to the first detection of a coetary amino acid.

  17. DNA typing by capillary electrophoresis

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, N.

    1997-10-08

    Capillary electrophoresis is becoming more and more important in nucleic acid analysis including DNA sequencing, typing and disease gene measurements. This work summarized the background of DNA typing. The recent development of capillary electrophoresis was also discussed. The second part of the thesis showed the principle of DNA typing based on using the allelic ladder as the absolute standard ladder in capillary electrophoresis system. Future work will be focused on demonstrating DNA typing on multiplex loci and examples of disease diagnosis in the on-line format of PCR-CE. Also capillary array electrophoresis system should allow high throughput, fast speed DNA typing. Only the introduction and conclusions for this report are available here. A reprint was removed for separate processing.

  18. Ex-dormant comets that come back to life: a search of reactivated comets

    Science.gov (United States)

    Quan-Zhi, YE

    2017-10-01

    Dormant or near-dormant short-period comets can unexpectedly regain the ability to eject dust. In many known cases, the resurrection is short-lived and lasts less than one orbit. However, it is possible that some resurrected comets can remain active in later perihelion passages. Here we conduct a search in the archival images of various facilities to look for these “reactivated” comets. We identify two candidates, 297P/Beshore and 332P/Ikeya-Murakami, both of which were found to be inactive or weakly active in the previous orbit before their discovery. We derive a reactivation rate of 0.007 events per comet per orbit, which implies that typical short-period comets only become temporarily dormant a few times or less. Smaller comets are prone to rotational instability and may undergo temporary dormancy more frequently. Next generation high-cadence surveys may find more reactivation events of these comets.

  19. Comet mission hopes to uncover Earth's origins

    CERN Multimedia

    Henderson, M

    2004-01-01

    "A European spacecraft that will hunt down a comet in search of clues to the origin of life on Earth will blast off tomorrow from the Kourou spaceport in French Guiana. The Rosetta probe will take 12 years to catch up with Churyumov-Gerasimenko before becoming the first spacecraft to make a soft, controlled landing on a comet's nucleus" (1 page).

  20. Meteoroid Streams from Sunskirter Comet Breakup

    Science.gov (United States)

    Jenniskens, P. M.

    2012-12-01

    In its first year of operations, the CAMS project (Cameras for Allsky Meteor Surveillance) has measured 47,000 meteoroid orbits at Earth, including some that pass the Sun as close as 0.008 AU. The population density increases significantly above perihelion distance q = 0.037 AU. Meteoroid streams are known with q about 0.1 AU. The Sun has a profound effect on comets that pass at 0.04-0.16 AU distance, called the sunskirter comets. SOHO and STEREO see families of small comets called the Marsden and Kracht groups. Sunlight is efficiently scattered by small 10-m sized fragments, making those fragments visible even when far from Earth. These comet groups are associated with meteor showers on Earth, in particular the Daytime Arietids and Delta Aquariids. All are related to 96P/Machholz, a highly inclined short-period (5.2 year) Jupiter family comet that comes to within 0.12 AU from the Sun, the smallest perihelion distance known among numbered comets. The proximity of the Sun speeds up the disintegration process, providing us a unique window on this important decay mechanism of Jupiter family comets and creating meteoroid streams. These are not the only sunskirting comets, however. In this presentation, we will present CAMS observations of the complete low-q meteoroid population at Earth and review their association with known parent bodies.

  1. RFP for the Comet Nuclei Tour (CONTOUR)

    DEFF Research Database (Denmark)

    Jørgensen, John Leif; Madsen, Peter Buch; Betto, Maurizio

    1999-01-01

    This document describes the ASC Star Tracker (performance, functionality, requirements etc.) to The Johns Hopkins University - Applied Physics Laboratory for their Comet Nuclei TOUR (CONTOUR) Program.......This document describes the ASC Star Tracker (performance, functionality, requirements etc.) to The Johns Hopkins University - Applied Physics Laboratory for their Comet Nuclei TOUR (CONTOUR) Program....

  2. The Geology of Comet 19/P Borrelly

    Science.gov (United States)

    Britt, D. T.; Boice, D. C; Buratti, B. J.; Hicks, M. D.; Nelson, R. M.; Oberst, J.; Sandel, B. R.; Soderblom, L. A.; Stern, S. A.; Thomas, N.

    2002-01-01

    The Deep Space One spacecraft flew by Comet 19P/Borrelly on September 22, 2001 and returned a rich array of imagery with resolutions of up to 48 m/pixel. These images provide a window into the surface structure, processes, and geological history of a comet. Additional information is contained in the original extended abstract.

  3. Phobos and Deimos: A Potential Comet Connection

    Science.gov (United States)

    Lee, P.

    2017-07-01

    Phobos and Deimos are similar in size and spectrum to an exceptionally large and spectrally uncommon asteroid among near-Earth objects: 3552 Don Quixote, a D-type NEO that's a comet. The martian moons' potential link to comets is discussed.

  4. Comets and How to Observe Them

    CERN Document Server

    Schmude, Richard

    2010-01-01

    Comets have inspired wonder, excitement and even fear ever since they were first observed. They contain material from early in the life of the Solar System, held in deep-freeze. This makes them key in our understanding of the formation and evolution of many Solar System bodies. Recent ground- and space-based observations have changed much in our understanding of comets. Comets and How to Observe Them gives a summary of our current knowledge and describes how amateur astronomers can contribute to the body of scientific knowledge of comets. This book contains many practical examples of how to construct comet light-curves, measure how fast a comet’s coma expands, and determine the rotation period of the nucleus. All these examples are illustrated with drawings and photographs.

  5. Origin of Short-Perihelion Comets

    Science.gov (United States)

    Guliyev, A. S.

    2011-01-01

    New regularities for short-perihelion comets are found. Distant nodes of cometary orbits of Kreutz family are concentrated in a plane with ascending node 76 and inclination 267 at the distance from 2 up to 3 a.u. and in a very narrow interval of longitudes. There is a correlation dependence between q and cos I concerning the found plane (coefficient of correlation 0.41). Similar results are received regarding to cometary families of Meyer, Kracht and Marsden. Distant nodes of these comets are concentrated close three planes (their parameters are discussed in the article) and at distances 1.4; 0.5; 6 a.u. accordingly. It is concluded that these comet groups were formed as a result of collision of parent bodies with meteoric streams. One more group, consisting of 7 comets is identified. 5 comet pairs are selected among sungrazers.

  6. Oort spike comets with large perihelion distances

    Science.gov (United States)

    Królikowska, Małgorzata; Dybczyński, Piotr A.

    2017-12-01

    The complete sample of large-perihelion nearly-parabolic comets discovered during the period 1901-2010 is studied, starting with their orbit determination. Next, an orbital evolution that includes three perihelion passages (previous-observed-next) is investigated in which a full model of Galactic perturbations and perturbations from passing stars is incorporated. We show that the distribution of planetary perturbations suffered by actual large-perihelion comets during their passage through the Solar system has a deep, unexpected minimum around zero, which indicates a lack of 'almost unperturbed' comets. Using a series of simulations we show that this deep well is moderately resistant to some diffusion of the orbital elements of the analysed comets. It seems reasonable to assert that the observed stream of these large-perihelion comets experienced a series of specific planetary configurations when passing through the planetary zone. An analysis of the past dynamics of these comets clearly shows that dynamically new comets can appear only when their original semimajor axes are greater than 20 000 au. On the other hand, dynamically old comets are completely absent for semimajor axes longer than 40 000 au. We demonstrate that the observed 1/aori-distribution exhibits a local minimum separating dynamically new from dynamically old comets. Long-term dynamical studies reveal a wide variety of orbital behaviour. Several interesting examples of the action of passing stars are also described, in particular the impact of Gliese 710, which will pass close to the Sun in the future. However, none of the obtained stellar perturbations is sufficient to change the dynamical status of the analysed comets.

  7. Comet Hyakutake's close encounter with the Sun

    Science.gov (United States)

    1996-05-01

    LASCO is a joint project between NRL (USA), the Max Planck Institut für Aeronomie (Germany), the Laboratoire d'Astronomie Spatiale (France), and the School of Physics and Space Research at the University of Birmingham (UK). "Such observations require a special instrument in space to suppress the glare of the Sun and reveal the comet and its tails", said Dr. Guenter Brueckner, NRL's principal investigator for LASCO. Scattering of sunlight in the Earth's atmosphere prevented good views from the ground during the comet's "perihelion passage" when it was closest to the Sun. The orbital period of Comet Hyakutake has been estimated to be 10,000 years. Hyakutake is called a "new" comet because it was not seen when, and if, it last visited the solar system. As Hyakutake approaches the Sun, it is being heated enormously. If this is the first visit of the comet, it could be broken into pieces, according to scientists. Images captured by the LASCO instrument have shown that this did not happen when the comet was in LASCO's field of view, which is approximately the size of the constellation Orion. "Comet Hyakutake could have passed through the solar system many times before", said Dr. Brueckner, who is also head of the NRL's Solar Physics Branch. "How many times remains a mystery". Hyakutake's orbit carries it back into the so-called "Oort Cloud" a vast collection of billions of comets that is located 1.4 light years away from the solar system. These comets are presumably the remnants of the cloud from which the solar system were formed billions of years ago. When the comet enters the outer atmosphere of the Sun, it begins to react with the Sun's environment and can be used as a "probe" of the solar corona. LASCO images show the head of the comet, and clearly visible are three separate tails that behave differently as Hyakutake swings around the Sun. These tails are made of different materials ; dust of different sizes, perhaps chunks of ice and atomic particles, each of

  8. On the reflectance of dust in comets

    Science.gov (United States)

    Zubko, Evgenij; Videen, Gorden; Shkuratov, Yuriy; Hines, Dean C.

    2017-11-01

    Reflectance of cometary dust is a key parameter used in the characterization of comets. In the literature, the reflectance of single-scattering cometary dust particles is widely assumed to be the same as that of the cometary nucleus. We discuss this assumption and demonstrate its inconsistency with photometric observations of comets, laboratory optical measurements, and numerical simulation of light scattering from single-scattering dust particles and particulate surfaces composed of the same particles. We estimate the reflectance of cometary dust particles using a comprehensive physical model of polarization measured in comets over wide range of phase angle and at different wavelengths in the visible. The model predicts that the reflectance of dust in comets inversely correlates with their maximum of positive polarization Pmax. We find that even the darkest dust particles appearing in comets with the highest Pmax, reflect considerably more incident solar-radiation energy, up to 200%, compared to what is thought for cometary nuclei. We also find that the reflectance retrieved from polarimetry in the visible appears in good quantitative accordance with previous estimations from infrared observations of comets. Our findings suggest that the dust production of comets is currently overestimated and may require revision.

  9. The Comet Halley Handbook: An Observer's Guide. Second Edition.

    Science.gov (United States)

    Yeomans, Donald K.

    This handbook contains information on: (1) the orbit of comet Halley; (2) the expected physical behavior of comet Halley in 1985-1986, considering brightness estimates, coma diameters, and tail lengths; (3) observing conditions for comet Halley in 1985-1986; and (4) observing conditions for the dust tail of comet Halley in 1985-1986. Additional…

  10. Comet Halley Returns. A Teacher's Guide, 1985-1986.

    Science.gov (United States)

    Chapman, Robert D.; Bondurant, R. Lynn, Jr.

    This booklet was designed as an aid for elementary and secondary school teachers. It is divided into two distinct parts. Part I is a brief tutorial which introduces some of the most important concepts about comets. Areas addressed include: the historical importance of Comet Halley; how comets are found and names; cometary orbits; what Comet Halley…

  11. COLOR SYSTEMATICS OF COMETS AND RELATED BODIES

    Energy Technology Data Exchange (ETDEWEB)

    Jewitt, David, E-mail: jewitt@ucla.edu [Department of Earth, Planetary and Space Sciences, UCLA, 595 Charles Young Drive East, Los Angeles, CA 90095-1567 (United States)

    2015-12-15

    Most comets are volatile-rich bodies that have recently entered the inner solar system following long-term storage in the Kuiper belt and the Oort cloud reservoirs. These reservoirs feed several distinct, short-lived “small body” populations. Here, we present new measurements of the optical colors of cometary and comet-related bodies including long-period (Oort cloud) comets, Damocloids (probable inactive nuclei of long-period comets) and Centaurs (recent escapees from the Kuiper belt and precursors to the Jupiter family comets). We combine the new measurements with published data on short-period comets, Jovian Trojans and Kuiper belt objects to examine the color systematics of the comet-related populations. We find that the mean optical colors of the dust in short-period and long-period comets are identical within the uncertainties of measurement, as are the colors of the dust and of the underlying nuclei. These populations show no evidence for scattering by optically small particles or for compositional gradients, even at the largest distances from the Sun, and no evidence for ultrared matter. Consistent with earlier work, ultrared surfaces are common in the Kuiper belt and on the Centaurs, but not in other small body populations, suggesting that this material is hidden or destroyed upon entry to the inner solar system. The onset of activity in the Centaurs and the disappearance of the ultrared matter in this population begin at about the same perihelion distance (∼10 AU), suggesting that the two are related. Blanketing of primordial surface materials by the fallback of sub-orbital ejecta, for which we calculate a very short timescale, is the likely mechanism. The same process should operate on any mass-losing body, explaining the absence of ultrared surface material in the entire comet population.

  12. A Post-Stardust Mission View of Jupiter Family Comets

    Science.gov (United States)

    Zolensky, M.

    2011-01-01

    Before the Stardust Mission, many persons (including the mission team) believed that comet nuclei would be geologically boring objects. Most believed that comet nucleus mineralogy would be close or identical to the chondritic interplanetary dust particles (IDPs), or perhaps contain mainly amorphous nebular condensates or that comets might even be composed mainly of preserved presolar material [1]. Amazingly, the results for Comet Wild 2 (a Jupiter class comet) were entirely different. Whether this particular comet will ultimately be shown to be typical or atypical will not be known for a rather long time, so we describe our new view of comets from the rather limited perspective of this single mission.

  13. A Comet Engulfs Mars: MAVEN Observations of Comet Siding Spring's Influence on the Martian Magnetosphere

    Science.gov (United States)

    Espley, Jared R.; Dibraccio, Gina A.; Connerney, John E. P.; Brain, David; Gruesbeck, Jacob; Soobiah, Yasir; Halekas, Jasper S.; Combi, Michael; Luhmann, Janet; Ma, Yingjuan

    2015-01-01

    The nucleus of comet C/2013 A1 (Siding Spring) passed within 141,000?km of Mars on 19 October 2014. Thus, the cometary coma and the plasma it produces washed over Mars for several hours producing significant effects in the Martian magnetosphere and upper atmosphere. We present observations from Mars Atmosphere and Volatile EvolutioN's (MAVEN's) particles and field's instruments that show the Martian magnetosphere was severely distorted during the comet's passage. We note four specific major effects: (1) a variable induced magnetospheric boundary, (2) a strong rotation of the magnetic field as the comet approached, (3) severely distorted and disordered ionospheric magnetic fields during the comet's closest approach, and (4) unusually strong magnetosheath turbulence lasting hours after the comet left. We argue that the comet produced effects comparable to that of a large solar storm (in terms of incident energy) and that our results are therefore important for future studies of atmospheric escape, MAVEN's primary science objective.

  14. Comet 'Bites the Dust' Around Dead Star

    Science.gov (United States)

    2006-01-01

    [figure removed for brevity, see original site] Infrared Spectrometer Graph This artist's concept illustrates a comet being torn to shreds around a dead star, or white dwarf, called G29-38. NASA's Spitzer Space Telescope observed a cloud of dust around this white dwarf that may have been generated from this type of comet disruption. The findings suggest that a host of other comet survivors may still orbit in this long-dead solar system. The white dwarf G29-38 began life as a star that was about three times as massive as our sun. Its death involved the same steps that the sun will ultimately undergo billions of years from now. According to theory, the G29-38 star became brighter and brighter as it aged, until it bloated up into a dying star called a red giant. This red giant was large enough to engulf and evaporate any terrestrial planets like Earth that happened to be in its way. Later, the red giant shed its outer atmosphere, leaving behind a shrunken skeleton of star, called a white dwarf. If the star did host a planetary system, outer planets akin to Jupiter and Neptune and a remote ring of icy comets would remain. The Spitzer observations provide observational evidence for this orbiting outpost of comet survivors. Astronomers speculate that one such comet was knocked into the inner regions of G29-38, possibly by an outer planet. As the comet approached very close to the white dwarf, it may have been torn apart by the star's tidal forces. Eventually, all that would be left of the comet is a disk of dust. This illustration shows a comet in the process of being pulverized: part of it still exists as a chain of small clumps, while the rest has already spread out into a dusty disk. Comet Shoemaker-Levy 9 broke apart in a similar fashion when it plunged into Jupiter in 1994. Evidence for Comets Found in Dead Star's Dust The graph of data, or spectrum, from NASA's Spitzer Space Telescope indicates that a dead star, or white dwarf, called G29-38, is shrouded by a cloud

  15. A New Orbit for Comet C/1865 B1 (Great Southern Comet of 1865)

    Science.gov (United States)

    Branham, Richard L., Jr.

    2018-04-01

    Comet C/1865 B1 (Great southern comet of 1865), observed only in the southern hemisphere, is one of a large number of comets with parabolic orbits. Given that there are 202 observations in right ascension and 165 in declination it proves possible to calculate a better orbit than that Körber published in 1887, the orbit used in various catalogs and data bases. C/1865 B1's orbit is hyperbolic and statistically distinguishable from a parabola. This object, therefore, cannot be considered an NEO. The comet has a small perihelion distance of 0.026 AU.

  16. A Comet Surface Sample Return System Project

    Data.gov (United States)

    National Aeronautics and Space Administration — The proposed Phase I investigation will focus on the development of spacecraft systems required to obtain a sample from the nucleus of a comet, hermetically seal the...

  17. Laboratory Study on Disconnection Events in Comets

    Science.gov (United States)

    Li, Yan-Fei; Li, Yu-Tong; Wang, Wei-Min; Yuan, Da-Wei; et al.

    2018-01-01

    When comets interacting with solar wind, straight and narrow plasma tails will be often formed. The most remarkable phenomenon of the plasma tails is the disconnection event, in which a plasma tail is uprooted from the comet's head and moves away from the comet. In this paper, the interaction process between a comet and solar wind is simulated by using a laser-driven plasma cloud to hit a cylinder obstacle. A disconnected plasma tail is observed behind the obstacle by optical shadowgraphy and interferometry. Our particle-in-cell simulations show that the diference in thermal velocity between ions and electrons induces an electrostatic field behind the obstacle. This field can lead to the convergence of ions to the central region, resulting in a disconnected plasma tail. This electrostatic field-induced model may be a possible explanation for the disconnection events of cometary tails.

  18. Radio Observations of Organics in Comets

    Science.gov (United States)

    Milam, Stefanie N.; Charnley, Steven B.; Kuan, Yi-Jehng; Chuang, Yo-Ling; Villanueva, Geronimo; Coulson, Iain; Remijan, Anthony J.

    2012-01-01

    A major observational challenge in cometary science is to quantify the extent to which chemical compounds can be linked to either interstellar or nebular chemistry. Recently, there have been complimentary observations from multiple facilities to try to unravel the chemical complexity of comets and their origins. Incorporating results from various techniques can gain further insight into the abundances, production rates, distributions, and formation mechanisms of molecules in these objects [I]. Such studies have provided great detail towards molecules with a-typical chemistries, such as H2CO [2]. We report multiwavelength spectral observations of comets from two dynamical families including the JFC 103P/Hartley 2 and a long period comet C/2009 PI (Garradd) with the Arizona Radio Observatory's SMT and 12-m telescopes, as well as the NRAO Greenbank telescope, and the James Clerk Maxwell Telescope. Multiple parent volatiles (e.g. HCN, CH30H, CO) as well as daughter products (e.g, CS and 01-1) have been detected in these objects. We will present a comparison of molecular abundances in these comets to those observed in others, supporting a long-term effort of building a comet taxonomy based on composition. Previous work has revealed a range of abundances of parent species (from "organics-poor" to "organics-rich") with respect to water among comets [3,4,5], however the statistics are not well constrained.

  19. Primary Separation: 2-D Electrophoresis

    NARCIS (Netherlands)

    Pedreschi Plasencia, R.P.

    2013-01-01

    The advancements in two-dimensional electrophoresis (2DE) have consolidated it as a key tool for gel-based proteomics applications. Nowadays, 2DE is extensively applied and it is a useful technique for the simultaneous separation of hundreds to thousands of proteins, analysis, and differential

  20. The Comet Radar Explorer Mission

    Science.gov (United States)

    Asphaug, Erik; Belton, Mike; Bockelee-Morvan, Dominique; Chesley, Steve; Delbo, Marco; Farnham, Tony; Gim, Yonggyu; Grimm, Robert; Herique, Alain; Kofman, Wlodek; Oberst, Juergen; Orosei, Roberto; Piqueux, Sylvain; Plaut, Jeff; Robinson, Mark; Sava, Paul; Heggy, Essam; Kurth, William; Scheeres, Dan; Denevi, Brett; Turtle, Elizabeth; Weissman, Paul

    2014-11-01

    Missions to cometary nuclei have revealed major geological surprises: (1) Global scale layers - do these persist through to the interior? Are they a record of primary accretion? (2) Smooth regions - are they landslides originating on the surface? Are they cryovolcanic? (3) Pits - are they impact craters or sublimation pits, or rooted in the interior? Unambiguous answers to these and other questions can be obtained by high definition 3D radar reflection imaging (RRI) of internal structure. RRI can answer many of the great unknowns in planetary science: How do primitive bodies accrete? Are cometary nuclei mostly ice? What drives their spectacular activity and evolution? The Comet Radar Explorer (CORE) mission will image the detailed internal structure of the nucleus of 10P/Tempel 2. This ~16 x 8 x 7 km Jupiter Family Comet (JFC), or its parent body, originated in the outer planets region possibly millions of years before planet formation. CORE arrives post-perihelion and observes the comet’s waning activity from safe distance. Once the nucleus is largely dormant, the spacecraft enters a ~20-km dedicated Radar Mapping Orbit (RMO). The exacting design of the RRI experiment and the precise navigation of RMO will achieve a highly focused 3D radar reflection image of internal structure, to tens of meters resolution, and tomographic images of velocity and attenuation to hundreds of meters resolution, tied to the gravity model and shape. Visible imagers will produce maps of the surface morphology, albedo, color, texture, and photometric response, and images for navigation and shape determination. The cameras will also monitor the structure and dynamics of the coma, and its dusty jets, allowing their correlation in 3D with deep interior structures and surface features. Repeated global high-resolution thermal images will probe the near-surface layers heated by the Sun. Derived maps of thermal inertia will be correlated with the radar boundary response, and photometry and

  1. Transport and Capture of Comets

    Science.gov (United States)

    Ross, S. D.; Koon, W. S.; Lo, M. W.; Marsden, J. E.

    2001-11-01

    The dynamics of comets and other solar system objects which have a three-body energy close to that of the collinear libration points are known to exhibit a complicated array of behaviors such as rapid transition between the interior and exterior Hill's regions, temporary capture, and collision. The invariant manifold structures of the collinear libration points for the restricted three-body problem, which exist for a range of energies, provide the framework for understanding these transport phenomena from a geometric point of view. In particular, the stable and unstable invariant manifold "tubes" associated to libration point orbits are the phase space conduits transporting material to and from the smaller primary body (e.g., Jupiter), and between primary bodies for separate three-body systems (e.g., Saturn and Jupiter). This point of view has worked well in describing the planar circular restricted three-body problem. The current work seeks to extend the results to three degrees of freedom. This work was supported by the National Science Foundation Grant No. KDI/ATM-9873133 under a contract with the Jet Propulsion Laboratory, NASA.

  2. Comet Tempel 1 Went Back to Sleep

    Science.gov (United States)

    2005-07-01

    Astronomers Having Used ESO Telescopes Start Analysing Unique Dataset on the Comet Following the Deep Impact Mission Ten days after part of the Deep Impact spacecraft plunged onto Comet Tempel 1 with the aim to create a crater and expose pristine material from beneath the surface, astronomers are back in the ESO Offices in Santiago, after more than a week of observing at the ESO La Silla Paranal Observatory. In this unprecedented observing campaign - among the most ambitious ever conducted by a single observatory - the astronomers have collected a large amount of invaluable data on this comet. The astronomers have now started the lengthy process of data reduction and analysis. Being all together in a single place, and in close contacts with the space mission' scientific team, they will try to assemble a clear picture of the comet and of the impact. The ESO observations were part of a worldwide campaign to observe this unique experiment. During the campaign, ESO was connected by phone, email, and videoconference with colleagues in all major observatories worldwide, and data were freely exchanged between the different groups. This unique collaborative spirit provides astronomers with data taken almost around the clock during several days and this, with the largest variety of instruments, making the Deep Impact observing campaign one of the most successful of its kind, and thereby, ensuring the greatest scientific outcome. From the current analysis, it appears most likely that the impactor did not create a large new zone of activity and may have failed to liberate a large quantity of pristine material from beneath the surface. ESO PR Photo 22/05 ESO PR Photo 22/05 Evolution of Comet Tempel 1 (FORS2/VLT) [Preview - JPEG: 400 x 701 pix - 128k] [Normal - JPEG: 800 x 1401 pix - 357k] ESO PR Photo 22/05 Animated Gif Caption: ESO PR Photo 22/05 shows the evolution of Comet Tempel 1 as observed with the FORS2 instrument on Antu (VLT). The images obtained at the VLT show that

  3. Enzyme assays.

    Science.gov (United States)

    Brodelius, P E

    1991-02-01

    The past year or so has seen the development of new enzyme assays, as well as the improvement of existing ones. Assays are becoming more rapid and sensitive as a result of modifications such as amplification of the enzyme product(s). Recombinant DNA technology is now being recognized as a particularly useful tool in the search for improved assay systems.

  4. A GREAT search for Deuterium in Comets

    Science.gov (United States)

    Mumma, Michael

    2013-10-01

    Comets are understood to be the most pristine bodies in the Solar System. Their compositions reflect the chemical state of materials at the very earliest evolutionary stages of the protosolar nebula and, as such, they provide detailed insight into the physical and chemical processes operating in planet-forming disks. Isotopic fractionation ratios of the molecular ices in the nucleus are regarded as signatures of formation processes. These ratios provide unique information on the natal heritage of those ices, and can also test the proposal that Earth's water and other volatiles were delivered by cometary bombardment. Measurement of deuterium fractionation ratios is thus a major goal in contemporary cometary science and the D/H ratio of water - the dominant volatile in comets - holds great promise for testing the formation history of cometary matter. The D/H ratio in cometary water has been measured in only eight comets. Seven were from the Oort Cloud reservoir and the D/H ratio was about twice that of the Earth's oceans. However, the recent Herschel measurement of HDO/H2O in 103P/Hartley-2 (the first from the Kuiper Belt) was consistent with exogenous delivery of Earth's water by comets. Outstanding questions remain: are cometary HDO/H2O ratios consistent with current theories of nebular chemical evolution or with an interstellar origin? Does the HDO/H2O ratio vary substantially among comet populations? Hartley-2 is the only Kuiper Belt comet with measured HDO/H2O, are there comets with similar ratios in the Oort cloud? These questions can only be addressed by measuring HDO/H2O ratios in many more suitable bright comets. We therefore propose to measure the D/H ratio in water in a suitable target-of-opportunity comet by performing observations of HDO and OH with the GREAT spectrometer on SOFIA. A multi-wavelength, ground-based observing campaign will also be conducted in support of the airborne observations.

  5. Abundant Solar Nebula Solids in Comets

    Science.gov (United States)

    Messenger, S.; Keller, L. P.; Nakamura-Messenger, K.; Nguyen, A. N.; Clemett, S.

    2016-01-01

    Comets have been proposed to consist of unprocessed interstellar materials together with a variable amount of thermally annealed interstellar grains. Recent studies of cometary solids in the laboratory have shown that comets instead consist of a wide range of materials from across the protoplanetary disk, in addition to a minor complement of interstellar materials. These advances were made possible by the return of direct samples of comet 81P/Wild 2 coma dust by the NASA Stardust mission and recent advances in microscale analytical techniques. Isotopic studies of 'cometary' chondritic porous interplanetary dust particles (CP-IDPs) and comet 81P/Wild 2 Stardust samples show that preserved interstellar materials are more abundant in comets than in any class of meteorite. Identified interstellar materials include sub-micron-sized presolar silicates, oxides, and SiC dust grains and some fraction of the organic material that binds the samples together. Presolar grain abundances reach 1 weight percentage in the most stardust-rich CP-IDPs, 50 times greater than in meteorites. Yet, order of magnitude variations in presolar grain abundances among CP-IDPs suggest cometary solids experienced significant variations in the degree of processing in the solar nebula. Comets contain a surprisingly high abundance of nebular solids formed or altered at high temperatures. Comet 81P/Wild 2 samples include 10-40 micron-sized, refractory Ca- Al-rich inclusion (CAI)-, chondrule-, and ameboid olivine aggregate (AOA)-like materials. The O isotopic compositions of these refractory materials are remarkably similar to their meteoritic counterparts, ranging from 5 percent enrichments in (sup 16) O to near-terrestrial values. Comet 81P/Wild 2 and CP-IDPs also contain abundant Mg-Fe crystalline and amorphous silicates whose O isotopic compositions are also consistent with Solar System origins. Unlike meteorites, that are dominated by locally-produced materials, comets appear to be composed of

  6. Migration of Interplanetary Dust and Comets

    Science.gov (United States)

    Ipatov, S. I.; Mather, J. C.

    Our studies of migration of interplanetary dust and comets were based on the results of integration of the orbital evolution of 15,000 dust particles and 30,000 Jupiter-family comets (JFCs) [1-3]. For asteroidal and cometary particles, the values of the ratio β between the radiation pressure force and the gravitational force varied from 1000 and 1 microns. The probability of a collision of a dust particle started from an asteroid or JFC with the Earth during a lifetime of the particle was maximum at diameter d ˜100 microns. For particles started from asteroids and comet 10P, this maximum probability was ˜0.01. Different studies of migration of dust particles and small bodies testify that the fraction of cometary dust particles of the overall dust population inside Saturn's orbit is considerable and can be dominant: (1) Cometary dust particles produced both inside and outside Jupiter's orbit are needed to explain the observed constant number density of dust particles at 3-18 AU. The number density of migrating trans-Neptunian particles near Jupiter's orbit is smaller by a factor of several than that beyond Saturn's orbit. Only a small fraction of asteroidal particles can get outside Jupiter's orbit. (2) Some (less than 0.1%) JFCs can reach typical near-Earth object orbits and remain there for millions of years. Dynamical lifetimes of most of the former JFCs that have typical near-Earth object orbits are about 106 -109 yr, so during most of these times they were extinct comets. Such former comets could disintegrate and produce a lot of mini-comets and dust. (3) Comparison of the velocities of zodiacal dust particles (velocities of MgI line) based on the distributions of particles over their orbital elements obtained in our runs [3-4] with the velocities obtained at the WHAM observations shows that only asteroidal dust particles cannot explain these observations, and particles produced by comets, including high-eccentricity comets, are needed for such explanation

  7. Polyacrylamide temperature gradient gel electrophoresis.

    Science.gov (United States)

    Viglasky, Viktor

    2013-01-01

    Temperature Gradient Gel Electrophoresis (TGGE) is a form of electrophoresis in which temperature gradient is used to denature molecules as they move through either acrylamide or agarose gel. TGGE can be applied to analyze DNA, RNA, protein-DNA complexes, and, less commonly, proteins. Separation of double-stranded DNA molecules during TGGE relies on temperature-dependent melting of the DNA duplex into two single-stranded DNA molecules. Therefore, the mobility of DNA reflects not only the size of the molecule but also its nucleotide composition, thereby allowing separation of DNA molecules of similar size with different sequences. Depending on the relative orientation of electric field and temperature gradient, TGGE can be performed in either a parallel or a perpendicular mode. The former is used to analyze multiple samples in the same gel, whereas the later allows detailed analysis of a single sample. This chapter is focused on analysis of DNA by polyacrylamide TGGE using the perpendicular mode.

  8. Ratcheted electrophoresis of Brownian particles

    Energy Technology Data Exchange (ETDEWEB)

    Kowalik, Mikołaj; Bishop, Kyle J. M., E-mail: kjmbishop@engr.psu.edu [Department of Chemical Engineering, The Pennsylvania State University, University Park, Pennsylvania 16802 (United States)

    2016-05-16

    The realization of nanoscale machines requires efficient methods by which to rectify unbiased perturbations to perform useful functions in the presence of significant thermal noise. The performance of such Brownian motors often depends sensitively on their operating conditions—in particular, on the relative rates of diffusive and deterministic motions. In this letter, we present a type of Brownian motor that uses contact charge electrophoresis of a colloidal particle within a ratcheted channel to achieve directed transport or perform useful work against an applied load. We analyze the stochastic dynamics of this model ratchet to show that it functions under any operating condition—even in the limit of strong thermal noise and in contrast to existing ratchets. The theoretical results presented here suggest that ratcheted electrophoresis could provide a basis for electrochemically powered, nanoscale machines capable of transport and actuation of nanoscale components.

  9. Evaluation of the internalization kinetics of the radiopharmaceutical {sup 99m}Tc-N{sub 2}S{sub 2}-Tat(49-57)Lys{sup 3}-Bn with diagnostic purposes, using comet assay; Evaluacion de la cinetica de internalizacion del radiofarmaco {sup 99m}Tc-N{sub 2}S{sub 2}-TAT(49-57)Lys{sup 3}-BN con fines diagnosticos, empleando ensayo cometa

    Energy Technology Data Exchange (ETDEWEB)

    Luna G, M. A.

    2011-07-01

    Gastrin-rea leasing peptide receptors (GRP-r) are over expressed in breast and prostate cancer cells. Bombesin (Bn) binds specifically and strongly to GRP-r and this is the base for to label the Bn with radionuclides by gamma rays. Tat (49-57) is a peptide that across the cell membrane easily so that, when it is conjugated to different proteins, it can works as a Trojan horse, facilitating the drug internalization to the cells. The radiopharmaceutical {sup 99m}Tc-N{sub 2}S{sub 2}-Tat(49-57)-Lys{sup 3}-Bn was prepared for diagnosis and therapy at early stage of breast cancer. The objective of this study was to determine the role of Tat in the internalization kinetics of radiopharmaceuticals measured by DNA damage induced by means of comet assay. Human lymphocytes were treated with the following protocols: a) Tat-Bn, b) {sup 99m}Tc-Bn, or c) {sup 99m}Tc-N{sub 2}S{sub 2}-Tat(49-57)-Lys{sup 3}-Bn, also an untreated group was conformed. The internalization was evaluated at 0, 5, 10, 15, 30 and 60 min after exposure with three repetitions each one, and for radiopharmaceuticals with 2.9, 6.6, 9.0 and 14.8 MBq activities. DNA damage was scored in 100 cells per time and treatment, as tail length and tail moment. A Kruskal-Wallis variance analysis with p{<=} 0.05 was applied for comparison between treatments. The results showed that the damage caused by {sup 99m}Tc-N{sub 2}S{sub 2}-Tat(49-57)-Lys{sup 3}-Bn is significantly higher than that caused by {sup 99m}Tc-Bn and Tat-Bn, showing that Tat favors the internalization of the radiopharmaceutical. (Author)

  10. DNA Sequencing by Capillary Electrophoresis

    Science.gov (United States)

    Karger, Barry L.; Guttman, Andras

    2009-01-01

    Sequencing of human and other genomes has been at the center of interest in the biomedical field over the past several decades and is now leading toward an era of personalized medicine. During this time, DNA sequencing methods have evolved from the labor intensive slab gel electrophoresis, through automated multicapillary electrophoresis systems using fluorophore labeling with multispectral imaging, to the “next generation” technologies of cyclic array, hybridization based, nanopore and single molecule sequencing. Deciphering the genetic blueprint and follow-up confirmatory sequencing of Homo sapiens and other genomes was only possible by the advent of modern sequencing technologies that was a result of step by step advances with a contribution of academics, medical personnel and instrument companies. While next generation sequencing is moving ahead at break-neck speed, the multicapillary electrophoretic systems played an essential role in the sequencing of the Human Genome, the foundation of the field of genomics. In this prospective, we wish to overview the role of capillary electrophoresis in DNA sequencing based in part of several of our articles in this journal. PMID:19517496

  11. MCDONALD OBSERVATORY FAINT COMET SPECTRO-PHOTOMETRIC SURVEY

    Data.gov (United States)

    National Aeronautics and Space Administration — The McDonald Observatory Faint Comet Survey data set presents spectral data from 152 observations of 17 comets taken using the Intensified Dissector Scanner...

  12. SURVEY OF COMET LIGHTCURVES V1.0

    Data.gov (United States)

    National Aeronautics and Space Administration — This is a dataset that compiles the data from various papers, which reported observations of brightness changes in comets that allowed to produce the comet...

  13. Dynamical evolution and disintegration of comets

    Science.gov (United States)

    Kresak, L.

    Current concepts of the origin and evolution of comets are reviewed. The place of their formation from which they have been delivered into the Oort reservoir is still an open problem, but the region of the outermost planets appears most probable. The interplay of stellar and planetary perturbations can be traced by model computations which reveal both the general trends and the variety of individual evolutionary paths. The present structure of the system of comets is controlled by the dynamical evolution of its individual members, limited by their physical aging by disintegration. Where the lifetimes are short, as in the Jupiter family of short-period comets, an equilibrium between elimination and replenishment is established. The role of different destructive processes and the resulting survival times are discussed.

  14. Comets As Objects of High Energy Astrophysics

    Science.gov (United States)

    Ibadov, S.

    2000-10-01

    Strong soft X-ray emission from comet Hyakutake C/1996 B2 was discovered with ROSAT in March 27, 1996 (Lisse et al. 1996, Science 274, 205-209) and the results of a theoretical approach (Ibadov 1990, Icarus 86, 283-288) served as a motive for that observations (Dennerl, Lisse and Truemper 1998, Private Communications). It is now well established that comets emit EUV and X-rays regularly (Dennerl, Englhauser and Truemper 1997, Science 277, 1625-1630; Dennerl 1998, Proc. 16th Int. Conf. Atomic Physics, Windsor, Ontario, Canada). To explain this phenomenon different theoretical models were proposed (Krasnopolsky 1997, Icarus 128, 365-385; Ibadov 1998, Proc. First XMM Workshop, Noordwijk, The Netherlands, and references therein). In the paper the problem of identifying X-ray generation mechanisms in comets will be considered.

  15. The mass disruption of Jupiter Family comets

    Science.gov (United States)

    Belton, Michael J. S.

    2015-01-01

    I show that the size-distribution of small scattered-disk trans-neptunian objects when derived from the observed size-distribution of Jupiter Family comets (JFCs) and other observational constraints implies that a large percentage (94-97%) of newly arrived active comets within a range of 0.2-15.4 km effective radius must physically disrupt, i.e., macroscopically disintegrate, within their median dynamical lifetime. Additional observational constraints include the numbers of dormant and active nuclei in the near-Earth object (NEO) population and the slope of their size distributions. I show that the cumulative power-law slope (-2.86 to -3.15) of the scattered-disk TNO hot population between 0.2 and 15.4 km effective radius is only weakly dependent on the size-dependence of the otherwise unknown disruption mechanism. Evidently, as JFC nuclei from the scattered disk evolve into the inner Solar System only a fraction achieve dormancy while the vast majority of small nuclei (e.g., primarily those with effective radius <2 km) break-up. The percentage disruption rate appears to be comparable with that of the dynamically distinct Oort cloud and Halley type comets (Levison, H.F., Morbidelli, A., Dones, L., Jedicke, R., Wiegert, P.A., Bottke Jr., W.F. [2002]. Science 296, 2212-2215) suggesting that all types of comet nuclei may have similar structural characteristics even though they may have different source regions and thermal histories. The typical disruption rate for a 1 km radius active nucleus is ∼5 × 10-5 disruptions/year and the dormancy rate is typically 3 times less. We also estimate that average fragmentation rates range from 0.01 to 0.04 events/year/comet, somewhat above the lower limit of 0.01 events/year/comet observed by Chen and Jewitt (Chen, J., Jewitt, D.C. [1994]. Icarus 108, 265-271).

  16. The use of 1986 Comet Halley observations for space projects

    Science.gov (United States)

    Kolyuka, Y. F.; Kudryavtsev, S. M.; Tarasov, V. P.; Tikhonov, V. F.

    1987-07-01

    The used observations of comet Halley in the current appearance for the improvement of the comet's orbit for purposes of the projects "Vega" and "Pathfinder" are described. The data on comet observations from different observatories and spacecraft "Vega" are analyzed. The improved orbit parameters are given.

  17. To Catch A Comet...Learning From Halley's.

    Science.gov (United States)

    National Aeronautics and Space Administration, Washington, DC.

    Comet chronicles and stories extend back over thousands of years. A common theme has been that comets are a major cause of catastrophe and tragedy here on earth. In addition, both Aristotle and Ptolemy believed that comets were phenomena within the earth's atmosphere, and it wasn't until the 16th century, when Danish astronomer Tycho Brache…

  18. Capillary electrophoresis theory and practice

    CERN Document Server

    Grossman, Paul D

    1992-01-01

    This book is designed to be a practical guide, used by wide audience, including those new to CE, those more experienced, routine users, those interested in technology development, and those involved with applications research. References have been emphasized to allow the reader to explore the detailed specifics and theoretical foundations.This book draws together the rapidly evolving, diverse, and multidisciplinary subject of capillary electrophoresis (CE). It is designed as a practical guide to be used by a wide audience, including those new to CE as well as more experienced users. T

  19. Spectral analysis of the Chandra comet survey

    NARCIS (Netherlands)

    Bodewits, D.; Christian, D. J.; Torney, M.; Dryer, M.; Lisse, C. M.; Dennerl, K.; Zurbuchen, T. H.; Wolk, S. J.; Tielens, A. G. G. M.; Hoekstra, R.

    Aims. We present results of the analysis of cometary X-ray spectra with an extended version of our charge exchange emission model (Bodewits et al. 2006). We have applied this model to the sample of 8 comets thus far observed with the Chandra X-ray observatory and acis spectrometer in the 300 - 1000

  20. Comet P/2001 T3 (Neat)

    Czech Academy of Sciences Publication Activity Database

    Lawrence, K. J.; Pravdo, S. H.; Helin, E. F.; Pravec, Petr; Kušnirák, Peter; Tichá, J.; Tichý, M.; Jelínek, P.

    č. 7733 (2001), s. 1 ISSN 0081-0304 R&D Projects: GA ČR GA205/99/0255 Institutional research plan: CEZ:AV0Z1003909 Keywords : comets * astrometry Subject RIV: BN - Astronomy, Celestial Mechanics, Astrophysics

  1. Comet P/2001 MD 7 (Linear)

    Czech Academy of Sciences Publication Activity Database

    Blythe, M.; Kotková, Lenka; Marsden, B. G.

    č. 7660 (2001), s. 1 ISSN 0081-0304 R&D Projects: GA ČR GA205/99/0255 Institutional research plan: CEZ:AV0Z1003909 Keywords : comets * astrometry Subject RIV: BN - Astronomy, Celestial Mechanics, Astrophysics

  2. Comet C/2001 Q6 (Neat)

    Czech Academy of Sciences Publication Activity Database

    Pravdo, S. H.; Helin, E. F.; Lawrence, K. J.; Tichý, M.; Kotková, Lenka; Wolf, M.; Balam, D.; Shelus, P. J.

    č. 7698 (2001), s. 1 ISSN 0081-0304 R&D Projects: GA ČR GA205/99/0255 Institutional research plan: CEZ:AV0Z1003909 Keywords : comets * astrometry Subject RIV: BN - Astronomy, Celestial Mechanics, Astrophysics

  3. Comet C/2001 A2 (Linear)

    Czech Academy of Sciences Publication Activity Database

    Pravec, Petr; Kotková, Lenka; Tichý, M.; Kočer, M.

    č. 7564 (2001), s. 1 ISSN 0081-0304 R&D Projects: GA ČR GA205/99/0255 Institutional research plan: CEZ:AV0Z1003909 Keywords : comets * astrometry Subject RIV: BN - Astronomy, Celestial Mechanics, Astrophysics

  4. Comet C/2001 Q1 (Neat)

    Czech Academy of Sciences Publication Activity Database

    Lawrence, K. J.; Helin, E. F.; Pravdo, S. H.; Pravec, Petr; Kušnirák, Peter; Kočer, M.; Spahr, T. B.

    č. 7685 (2001), s. 1 ISSN 0081-0304 R&D Projects: GA ČR GA205/99/0255 Institutional research plan: CEZ:AV0Z1003909 Keywords : comets * astrometry Subject RIV: BN - Astronomy, Celestial Mechanics, Astrophysics

  5. Comet C/2001 A1 (Linear)

    Czech Academy of Sciences Publication Activity Database

    Blythe, M.; Dawson, M.; Kornos, L.; Koleny, P.; Kotková, Lenka; Tichá, J.; Tichý, M.

    č. 7561 (2001), s. 1 ISSN 0081-0304 R&D Projects: GA ČR GA205/99/0255 Institutional research plan: CEZ:AV0Z1003909 Keywords : comets * astrometry Subject RIV: BN - Astronomy, Celestial Mechanics, Astrophysics

  6. COMETARY VOLATILES AND THE ORIGIN OF COMETS

    Energy Technology Data Exchange (ETDEWEB)

    A' Hearn, Michael F.; Feaga, Lori M.; Sunshine, Jessica M.; Besse, Sebastien; Bodewits, Dennis; Farnham, Tony L.; Kelley, Michael S. [Department of Astronomy, University of Maryland, College Park, MD 20742 (United States); Keller, H. Uwe [Institute for Geophysics and Extraterrestrial Physics, Technische Universitaet Braunschweig, D-38106 Braunschweig (Germany); Kawakita, Hideyo [Department of Physics, Kyoto Sangyo University, Kamigamo JP Kita-ku, Kyoto 603-8555 (Japan); Hampton, Donald L. [Geophysical Institute, University of Alaska Fairbanks, 903 Koyukuk Drive, Fairbanks, AK 99775 (United States); Kissel, Jochen [Max-Planck-Institut for Solar System Research, Max-Planck-Strasse 2, D-37191 Katlenburg-Lindau (Germany); Klaasen, Kenneth P.; Yeomans, Donald K. [Jet Propulsion Laboratory/Caltech, Pasadena, CA 91109 (United States); McFadden, Lucy A. [NASA Goddard Space Flight Center, Greenbelt, MD 20771 (United States); Meech, Karen J. [Institute for Astronomy, 2680 Woodlawn Drive, Honolulu, HI 96822 (United States); Schultz, Peter H. [Department of Geological Sciences, Brown University, Providence, RI 02912 (United States); Thomas, Peter C.; Veverka, Joseph [Department of Astronomy, Cornell University, Ithaca, NY 14853 (United States); Groussin, Olivier [Laboratoire d' Astrophysique de Marseille, Universite d' Aix-Marseille and CNRS, UMR7326, 38 rue F. Joliot-Curie, F-13388 Marseille Cedex 13 (France); Lisse, Carey M., E-mail: ma@astro.umd.edu [Space Department, JHU-APL, 11100 Johns Hopkins Road, Laurel, MD 20723 (United States); and others

    2012-10-10

    We describe recent results on the CO/CO{sub 2}/H{sub 2}O composition of comets together with a survey of older literature (primarily for CO/H{sub 2}O) and compare these with models of the protoplanetary disk. Even with the currently small sample, there is a wide dispersion in abundance ratios and little if any systematic difference between Jupiter-family comets (JFCs) and long-period and Halley-type comets (LPCs and HTCs). We argue that the cometary observations require reactions on grain surfaces to convert CO to CO{sub 2} and also require formation of all types of comets in largely, but not entirely, overlapping regions, probably between the CO and CO{sub 2} snow lines. Any difference in the regions of formation is in the opposite direction from the classical picture with the JFCs having formed closer to the Sun than the LPCs. In the classical picture, the LPCs formed in the region of the giant planets and the JFCs formed in the Kuiper Belt. However, these data suggest, consistent with suggestions on dynamical grounds, that the JFCs and LPCs formed in largely overlapping regions where the giant planets are today and with JFCs on average forming slightly closer to the Sun than did the LPCs. Presumably at least the JFCs passed through the scattered disk on their way to their present dynamical family.

  7. Comets and the origin and evolution of life

    CERN Document Server

    McKay, Christopher P

    2006-01-01

    Nine years after the publication of Comets and the Origin and Evolution of Life, one of the pioneering books in Astrobiology, this second edition revisits the role comets may have played in the origins and evolution of life. Recent analyses of Antarctic micrometeorites and ancient rocks in Australia and South Africa, the continuing progress in discovering complex organic macromolecules in comets, protostars and interstellar clouds, new insights into organic synthesis in comets, and numerical simulations of comet impacts on the Earth and other members of the solar system yield a spectacular wea

  8. Conducting polymer electrodes for gel electrophoresis.

    Directory of Open Access Journals (Sweden)

    Katarina Bengtsson

    Full Text Available In nearly all cases, electrophoresis in gels is driven via the electrolysis of water at the electrodes, where the process consumes water and produces electrochemical by-products. We have previously demonstrated that π-conjugated polymers such as poly(3,4-ethylenedioxythiophene (PEDOT can be placed between traditional metal electrodes and an electrolyte to mitigate electrolysis in liquid (capillary electroosmosis/electrophoresis systems. In this report, we extend our previous result to gel electrophoresis, and show that electrodes containing PEDOT can be used with a commercial polyacrylamide gel electrophoresis system with minimal impact to the resulting gel image or the ionic transport measured during a separation.

  9. Conducting polymer electrodes for gel electrophoresis.

    Science.gov (United States)

    Bengtsson, Katarina; Nilsson, Sara; Robinson, Nathaniel D

    2014-01-01

    In nearly all cases, electrophoresis in gels is driven via the electrolysis of water at the electrodes, where the process consumes water and produces electrochemical by-products. We have previously demonstrated that π-conjugated polymers such as poly(3,4-ethylenedioxythiophene) (PEDOT) can be placed between traditional metal electrodes and an electrolyte to mitigate electrolysis in liquid (capillary electroosmosis/electrophoresis) systems. In this report, we extend our previous result to gel electrophoresis, and show that electrodes containing PEDOT can be used with a commercial polyacrylamide gel electrophoresis system with minimal impact to the resulting gel image or the ionic transport measured during a separation.

  10. The Greatest Comets in History Broom Stars and Celestial Scimitars

    CERN Document Server

    Seargent, David A J

    2009-01-01

    Comets have fascinated and awed humankind since ancient times. Of the thousands of comets recorded throughout history, those deemed to have been the most spectacular have been described in the accounts of eyewitnesses and often recorded in official documents. This book introduces you to the greatest of the greats, starting with the comet in 372 B. C. called "Aristotle’s Comet" and ending with the spectacular appearance of McNaught’s Comet in 2007. There is an introductory chapter explaining what comets are and how they are classified, and correcting a few popular misconceptions. Later in the book you will read about the different returns of Halley’s Comet and the Kreutz sungrazing group, often called the kamikaze comets. There is even a chapter on comets that were visible in broad daylight. This book is unique. There are a few books on comets that make passing reference to some of the more famous or spectacular objects of the past, and a few catalogs with long lists of comets. But little detailed and de...

  11. Western blotting using capillary electrophoresis.

    Science.gov (United States)

    Anderson, Gwendolyn J; M Cipolla, Cynthia; Kennedy, Robert T

    2011-02-15

    A microscale Western blotting system based on separating sodium-dodecyl sulfate protein complexes by capillary gel electrophoresis followed by deposition onto a blotting membrane for immunoassay is described. In the system, the separation capillary is grounded through a sheath capillary to a mobile X-Y translation stage which moves a blotting membrane past the capillary outlet for protein deposition. The blotting membrane is moistened with a methanol and buffer mixture to facilitate protein adsorption. Although discrete protein zones could be detected, bands were broadened by ∼1.7-fold by transfer to membrane. A complete Western blot for lysozyme was completed in about one hour with 50 pg mass detection limit from low microgram per milliliter samples. These results demonstrate substantial reduction in time requirements and improvement in mass sensitivity compared to conventional Western blots. Western blotting using capillary electrophoresis shows promise to analyze low volume samples with reduced reagents and time, while retaining the information content of a typical Western blot.

  12. Recent advances of capillary electrophoresis in pharmaceutical analysis.

    Science.gov (United States)

    Suntornsuk, Leena

    2010-09-01

    This review covers recent advances of capillary electrophoresis (CE) in pharmaceutical analysis. The principle, instrumentation, and conventional modes of CE are briefly discussed. Advances in the different CE techniques (non-aqueous CE, microemulsion electrokinetic chromatography, capillary isotachophoresis, capillary electrochromatography, and immunoaffinity CE), detection techniques (mass spectrometry, light-emitting diode, fluorescence, chemiluminescence, and contactless conductivity), on-line sample pretreatment (flow injection) and chiral separation are described. Applications of CE to assay of active pharmaceutical ingredients (APIs), drug impurity testing, chiral drug separation, and determination of APIs in biological fluids published from 2008 to 2009 are tabulated.

  13. Long-period comets with non-gravitational effects

    Science.gov (United States)

    Królikowska, M.

    2004-12-01

    The influence of non-gravitational effects on the motion of long-period comets is analyzed. Investigations were performed for about 60 nearly-parabolic comets discovered during the last thirty years. All three non-gravitational parameters A1, A2, A3 were detected in the motion of 19 comets on the basis of positional observations only. For these comets the standard g(r) function as well as some of its modifications were used to model the ice sublimation from the cometary nucleus. For 14 of 19 comets the orientation of spin axes of the nucleus were also determined. Finally, the magnitude of the non-gravitational effects derived for the long-period comets have been compared with observed the non-gravitational perturbations on periodic comets. Tables 1, 2, 4 and 6 are only available in electronic form at http://www.edpsciences.org

  14. Unveiling the formation and evolution of comets

    Science.gov (United States)

    Lasue, J.; Levasseur-Regourd, A. C.; Botet, R.; Coradini, A.; Desanctis, M. C.; Kofman, W.

    2007-08-01

    Comet nuclei are considered as the most pristine bodies of the solar system and consequently their study sheds an important light on the processes occurring during the initial stages of the solar system formation. The analysis of the porosity and bulk density of such primordial bodies is especially important to understand their capacity to retain volatile components (organics and ices) present in the early solar nebula. Typical tensile strengths deduced for comet nuclei range from below 102N.m-2 from the Deep Impact mission [1] up to 104N.m-2 from the study of comet C/1999 S4 LINEAR breakup [2] and meteoroids [3]. A bulk density of about 350 kg/m3 has been obtained for 9P/Tempel 1 from the Deep Impact mission [4]. Moreover the properties of dust released from the comets strongly confirm such values. Instruments flying-by comet 1P/Halley had discovered the presence of organics, and pointed out the dust low albedo and extremely low density while analyses of Interplanetary Dust Particles collected in the stratosphere and remote spectroscopic observations have indicated that cometary dust consists of an un-equilibrated heterogeneous mixture of organic refractory materials and of amorphous and crystalline silicate minerals [5], as recently confirmed by Stardust [6]. Observations of the solar scattered light, together with elaborate simulations, give an estimation of the mass ratio between silicates and absorbing organics, the size distribution and the structure of the dust particles, suggesting that a fair amount consists in fluffy aggregates built up from submicronic grains [7,8], as recently confirmed by the analysis of dust craters and aerogel tracks on Stardust collector showing for some large particles (up to 100 μm) an extraordinary fluffy structure [9]. Simulations have been developed in our teams to describe the aspects of comet aggregation and evolution that have not been thoroughly explained yet. Particle aggregation simulations taking into account cohesive

  15. ESA Unveils Its New Comet Chaser.

    Science.gov (United States)

    1999-07-01

    The objective is to study one of these primordial objects at close quarters by placing a lander on its surface and chasing, with an orbiter, the comet for millions of kilometres through space. Comets - among the oldest (4.6 billion years!) and last altered objects in the solar system - are regarded as the building blocks from which the planets formed. Thus the Rosetta's discoveries will allow the scientists to learn more about birth and evolution of the planets and about the origin of life on the Earth. The final design of the Rosetta orbiter will be revealed for the first time at the Royal Society in London on 1 July when a 1:4 scale model will be unveiled by ESA's Director of Science, Prof.. Roger Bonnet. (The full size version of the spacecraft is 32 metres across, so large that it would stretch the entire width of a football pitch. Almost 90 of this is accounted for by the giant solar panels which are needed to provide electrical power in the dark depths of the Solar System). "Rosetta is a mission of major scientific importance," said Prof. Bonnet. "It will build on the discoveries made by Giotto and confirm ESA's leading role in the exploration of the Solar System and the Universe as a whole." The timing of this event has been chosen to coincide with the London meeting of the Rosetta Science Working Team and the second Earth flyby of the now non-operational Giotto spacecraft. In addition, the opening of the British Museum's 'Cracking Codes' Exhibition, for which the Rosetta Stone is the centrepiece, is set to take place on 10 July. The Rosetta mission. Rosetta is the third Cornerstone in ESA's 'Horizon 2000' long-term scientific programme. It will be launched by Ariane 5 rocket from Kourou spaceport in French Guiana in January 2003. In order to gain sufficient speed to reach the distant comet, Rosetta will require gravity assists from the Earth (twice) and Mars. After swinging around Mars in May 2005, Rosetta will return to Earth's vicinity in October 2005 and

  16. Adapting capillary gel electrophoresis as a sensitive, high-throughput method to accelerate characterization of nucleic acid metabolic enzymes

    OpenAIRE

    Greenough, Lucia; Schermerhorn, Kelly M.; Mazzola, Laurie; Bybee, Joanna; Rivizzigno, Danielle; Cantin, Elizabeth; Slatko, Barton E.; Gardner, Andrew F.

    2015-01-01

    Detailed biochemical characterization of nucleic acid enzymes is fundamental to understanding nucleic acid metabolism, genome replication and repair. We report the development of a rapid, high-throughput fluorescence capillary gel electrophoresis method as an alternative to traditional polyacrylamide gel electrophoresis to characterize nucleic acid metabolic enzymes. The principles of assay design described here can be applied to nearly any enzyme system that acts on a fluorescently labeled o...

  17. Reducing the Asteroid and Comet Impact Hazard

    Science.gov (United States)

    Crawford, David

    1998-10-01

    Of the 140 impact craters known on the surface of Earth, the most famous was created about 65 million years ago when a 10 km asteroid or comet came down in shallow water near the present day town of Chicxulub, Mexico. With a kinetic energy equivalent to 100 trillion tons of TNT, the impact event lofted enough debris onto globe-straddling trajectories to flash heat much of the surface of the Earth and then darken the skies for several years. Numerous investigations have demonstrated that such an event, which happens, on average, every 100 million years, caused extreme stress on Earth's climate and most likely led to the extinction of many species. Computational simulations demonstrate that more numerous asteroids or comets as small as 1 km in diameter, impacting, on average, every 300,000 years may be globally catastrophic. Indeed, the odds of an individual dying from a relatively frequent 1 km impacting object (about 1 in 10,000) are substantially greater than from the impact of an infrequent dinosaur killer (1 in 1,000,000). What can we do to reduce the hazard from impacting comets and asteroids? First, we should find what's out there with our name on it. Only about 10 percent of the potential Earth-crossing asteroids have been found. Even at the greatly increased detection rate of recent years, it will be several decades before we've found 90 percent of the Earth-crossers. Second, we should learn everything we can about the physical, compositional and mechanical properties of asteroids and comets. A recent computational study demonstrated that weakly bound asteroids (little more than rubble piles) are easier to break than deflect(E. Asphaug, S. J. Ostro, R. S. Hudson, D. J. Scheeres and W. Benz (1998), Nature, Vol. 393, pp. 437-440.). Is this an advantage or disadvantage? Third, we should study potential means of mitigating the hazard by deflecting the object while still in space or evacuating affected regions (such as coastlines) of the Earth. Because the

  18. Supramolecular gel electrophoresis of large DNA fragments.

    Science.gov (United States)

    Tazawa, Shohei; Kobayashi, Kazuhiro; Oyoshi, Takanori; Yamanaka, Masamichi

    2017-10-01

    Pulsed-field gel electrophoresis is a frequent technique used to separate exceptionally large DNA fragments. In a typical continuous field electrophoresis, it is challenging to separate DNA fragments larger than 20 kbp because they migrate at a comparable rate. To overcome this challenge, it is necessary to develop a novel matrix for the electrophoresis. Here, we describe the electrophoresis of large DNA fragments up to 166 kbp using a supramolecular gel matrix and a typical continuous field electrophoresis system. C 3 -symmetric tris-urea self-assembled into a supramolecular hydrogel in tris-boric acid-EDTA buffer, a typical buffer for DNA electrophoresis, and the supramolecular hydrogel was used as a matrix for electrophoresis to separate large DNA fragments. Three types of DNA marker, the λ-Hind III digest (2 to 23 kbp), Lambda DNA-Mono Cut Mix (10 to 49 kbp), and Marker 7 GT (10 to 165 kbp), were analyzed in this study. Large DNA fragments of greater than 100 kbp showed distinct mobility using a typical continuous field electrophoresis system. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  19. The Cutting Edge of Affinity Electrophoresis Technology

    Science.gov (United States)

    Kinoshita, Eiji; Kinoshita-Kikuta, Emiko; Koike, Tohru

    2015-01-01

    Affinity electrophoresis is an important technique that is widely used to separate and analyze biomolecules in the fields of biology and medicine. Both quantitative and qualitative information can be gained through affinity electrophoresis. Affinity electrophoresis can be applied through a variety of strategies, such as mobility shift electrophoresis, charge shift electrophoresis or capillary affinity electrophoresis. These strategies are based on changes in the electrophoretic patterns of biological macromolecules that result from interactions or complex-formation processes that induce changes in the size or total charge of the molecules. Nucleic acid fragments can be characterized through their affinity to other molecules, for example transcriptional factor proteins. Hydrophobic membrane proteins can be identified by means of a shift in the mobility induced by a charged detergent. The various strategies have also been used in the estimation of association/disassociation constants. Some of these strategies have similarities to affinity chromatography, in that they use a probe or ligand immobilized on a supported matrix for electrophoresis. Such methods have recently contributed to profiling of major posttranslational modifications of proteins, such as glycosylation or phosphorylation. Here, we describe advances in analytical techniques involving affinity electrophoresis that have appeared during the last five years. PMID:28248262

  20. Improvement of electrophoresis performance by spectral analysis ...

    African Journals Online (AJOL)

    This paper describes a new design of standard agarose gel electrophoresis procedure for nucleic acids analysis. The electrophoresis was improved by using the real-time spectral analysis of the samples to increase its performance. A laser beam illuminated the analysed sample at wavelength with the highest absorption of ...

  1. The Cutting Edge of Affinity Electrophoresis Technology.

    Science.gov (United States)

    Kinoshita, Eiji; Kinoshita-Kikuta, Emiko; Koike, Tohru

    2015-03-18

    Affinity electrophoresis is an important technique that is widely used to separate and analyze biomolecules in the fields of biology and medicine. Both quantitative and qualitative information can be gained through affinity electrophoresis. Affinity electrophoresis can be applied through a variety of strategies, such as mobility shift electrophoresis, charge shift electrophoresis or capillary affinity electrophoresis. These strategies are based on changes in the electrophoretic patterns of biological macromolecules that result from interactions or complex-formation processes that induce changes in the size or total charge of the molecules. Nucleic acid fragments can be characterized through their affinity to other molecules, for example transcriptional factor proteins. Hydrophobic membrane proteins can be identified by means of a shift in the mobility induced by a charged detergent. The various strategies have also been used in the estimation of association/disassociation constants. Some of these strategies have similarities to affinity chromatography, in that they use a probe or ligand immobilized on a supported matrix for electrophoresis. Such methods have recently contributed to profiling of major posttranslational modifications of proteins, such as glycosylation or phosphorylation. Here, we describe advances in analytical techniques involving affinity electrophoresis that have appeared during the last five years.

  2. Electrophoresis-mass spectrometry probe

    Science.gov (United States)

    Andresen, Brian D.; Fought, Eric R.

    1987-01-01

    The invention involves a new technique for the separation of complex mixtures of chemicals, which utilizes a unique interface probe for conventional mass spectrometers which allows the electrophoretically separated compounds to be analyzed in real-time by a mass spectrometer. This new chemical analysis interface, which couples electrophoresis with mass spectrometry, allows complex mixtures to be analyzed very rapidly, with much greater specificity, and with greater sensitivity. The interface or probe provides a means whereby large and/or polar molecules in complex mixtures to be completely characterized. The preferred embodiment of the probe utilizes a double capillary tip which allows the probe tip to be continually wetted by the buffer, which provides for increased heat dissipation, and results in a continually operating interface which is more durable and electronically stable than the illustrated single capillary tip probe interface.

  3. Capillary electrophoresis of proteins for proteomic studies.

    Science.gov (United States)

    Manabe, T

    1999-10-01

    Analyses of proteins in complex mixtures such as cell lyzates are presently performed mainly by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. For structural analysis, each protein in a spot is digested with proteases and the fragment peptides are subjected to Edman sequencing and/or mass spectrometry. These works aim at the total analysis of proteins in a complex mixture and reconstruction of their cooperative functions. Genomic studies are now being combined with these proteomic studies. This review article focuses on the application of capillary electrophoresis aiming at the total analysis of complex protein systems or structural analysis of each separated protein. From this viewpoint, articles on capillary zone electrophoresis, capillary isoelectric focusing, and sieving SDS capillary electrophoresis are reviewed. Since these techniques of capillary electrophoresis have been thoroughly reviewed previously, papers published in 1997 and 1998 are mainly covered.

  4. Research on pre-staining gel electrophoresis

    International Nuclear Information System (INIS)

    Zhong Ruibo; Liu Yushuang; Zhang Ping; Liu Jingran; Zhao Guofen; Zhang Feng

    2014-01-01

    Background: Gel electrophoresis is a powerful biochemical separation technique. Most biological molecules are completely transparent in the visible region of light, so it is necessary to use staining to show the results after gel electrophoresis, and the general steps of conventional staining methods are time-consuming. Purpose: We try to develop a novel approach to simplify the gel electrophoresis: Pre-Staining Gel Electrophoresis (PSGE), which can make the gel electrophoresis results monitored in real time. Methods: Pre-stain the protein samples with Coomassie Brilliant Blue (CBB) for 30 min before loading the sample into the gel well. Results and Conclusion: PSGE can be successfully used to analyze the binding efficiency of Bovine Serum Albumin (BSA) and amphiphilic polymer via chemical coupling and physical absorption, and the double PSGE also shows a great potential in bio-analytical chemistry. (authors)

  5. Ultrasound lung comets in extravascular fluid diagnostics

    Directory of Open Access Journals (Sweden)

    М. Н. Алехин

    2015-10-01

    Full Text Available The review is devoted to the analysis of ultrasound lung comets (ULCs that may be observed during ultrasound lung examination in cardiac patients in the case of accumulation of extravascular fluid in the lungs. Recent data on the mechanisms of these artefacts, examination and outcome evaluation methods are discussed. Differential diagnostics problems in patients with cardiac and pulmonary genesis of ULCs are also covered.

  6. EXTINCTION OF SPECIES BY PERIODIC COMET SHOWERS

    Energy Technology Data Exchange (ETDEWEB)

    Davis, M.; Hut, P.; Muller, R.A.

    1984-01-01

    A 26-Myr periodicity has recently been seen in the fossil record of extinction in the geological past. At least two of these extinctions are known to be associated with the impact on the Earth of a comet or asteroid with a diameter of a few kilometres. We propose that the periodic events are triggered by an unseen companion to the Sun, travelling in a moderately eccentric orbit, which at its closest approach (perihelion) passes through the 'Oort cloud' of comets which surrounds the Sun. During each passage this unseen solar companion perturbs the orbits of these comets, sending a large number of them (over 1 x 10{sup 9}) into paths which reach the inner Solar System. Several of these hit the Earth, on average, in the following million years. At present the unseen companion should be approximately at its maximum distance from the Sun, {approx}2.4 light yr, and it will present no danger to the Earth until approximately AD 15,000,000.

  7. Comets: Role and importance to exobiology

    Science.gov (United States)

    Delsemme, Armand H.

    1992-01-01

    The transfer of organic compounds from interstellar space to the outskirts of a protoplanetary disk, their accretion into cometary objects, and the transport of the latter into the inner solar system by orbital diffusion throw a new light on the central problem of exobiology. It suggests the existence of a cosmic mechanism, working everywhere, that can supply prebiotic compounds to ubiquitous rocky planets, in search of the proper environment to start life in many places in the Universe. Under the heading of chemistry of the cometary nucleus, the following topics are covered: radial homogeneity of the nucleus; the dust-to-ice ratio; nature of the dust grains; origin of the dust in comets; nature of the volatile fraction; the CO distribution in comet Halley; dust contribution to the volatile fraction; elemental balance sheet of comet Halley; quantitative molecular analysis of the volatile fraction; and isotopic ratios. Under the heading of exogenous origin of carbon on terrestrial planets the following topics are covered: evidence for a high-temperature phase; from planetesimals to planets; a veneer of volatile and organic material; and cometary contribution.

  8. Perihelion evolution of observed new comets implies the dominance of the galactic tide in making Oort cloud comets discernable

    Science.gov (United States)

    Matese, John J.; Lissauer, Jack J.

    2004-08-01

    For an Oort cloud comet to be seen as a new comet, its perihelion must be moved from a point exterior to the loss cylinder boundary to a point interior to observable limits in a single orbit. The galactic tide can do this continuously, in a non-impulsive manner. Near-parabolic comets, with specific angular momentum H∝ √q, will most easily be made observable. Therefore, to reduce the perihelion distance H must decrease. Since weakly perturbed comets are, in general, more numerous than strongly perturbed comets, we can anticipate that new comets made observable by a weak tidal torque will more likely be first observed when their slowly changing perihelion distances are approaching their minimum osculating values under the action of the tide, rather than receding from their minimum values. That is, defining Δ Htide as the vector change due to the galactic tidal torque during the prior orbit, and Hobs as the observed vector, the sign S≡Sign( Hobs·Δ Htide) will more likely be -1 than +1 if a weak galactic tidal perturbation indeed dominates in making comets observable. Using comet data of the highest quality class (1A) for new comets ( a>10,000 AU), we find that 49 comets have S=-1 and 22 have S=+1. The binomial probability that as many or more would exhibit this characteristic if in fact S=∓1 were equally likely is only 0.0009. This characteristic also persists in other long-period comet populations, lending support to the notion that they are dominated by comets recently arrived from the outer Oort cloud. The preponderance of S=-1 also correlates with weakly perturbed (i.e., smaller semimajor axis) new comets in a statistically significant manner. This is strong evidence that the data are of sufficiently high quality and sufficiently free of observational selection effects to detect this unique imprint of the tide.

  9. Investigation on accordance of DNA double-strand break of blood between in vivo and in vitro irradiation using single cell gel electrophoresis

    International Nuclear Information System (INIS)

    Liu Qiang; Jiang Enhai; Li Jin; Tang Weisheng; Wang Zhiquan; Zhao Yongcheng; Fan Feiyue

    2006-01-01

    Objective: To observe the consistency of DNA double-strand break between in vivo and in vitro irradiation, as a prophase study in radiation biodosimetry using single cell gel electrophoresis (SCGE). Methods: Detect DNA double-strand break after whole-body and in vitro radiation in mice lymphocytes using neutral single cell gel electrophoresis. The comet images were processed by CASP software and all the data were analysed by SPSS12.0. Results: There is no difference between in vivo and in vitro irradiation group in HDNA%, TDNA%, CL, TL, TM and OTM. Conclusion: The result of neutral single cell gel electrophoresis shortly after in vitro irradiation can precisely reflect the DNA double-strand break of lymphocytes in whole-body irradiation. (authors)

  10. Capillary electrophoresis and nanomaterials - Part I: Capillary electrophoresis of nanomaterials.

    Science.gov (United States)

    Adam, Vojtech; Vaculovicova, Marketa

    2017-10-01

    Nanomaterials are in analytical science used for a broad range of purposes, covering the area of sample pretreatment as well as separation, detection, and identification of target molecules. This part of the review covers capillary electrophoresis (CE) of nanomaterials and focuses on the application of CE as a method for characterization used during nanomaterial synthesis and modification as well as the monitoring of their properties and interactions with other molecules. The heterogeneity of the nanomaterial family is extremely large. Depending on different definitions of the term Nanomaterial/Nanoparticle, the group may cover metal and polymeric nanoparticles, carbon nanomaterials, liposomes and even dendrimers. Moreover, these nanomaterials are usually subjected to some kind of surface modification or functionalization, which broadens the diversity even more. Not only for purposes of verification of nanomaterial synthesis and batch-to-batch quality check, but also for determination the polydispersity and for functionality characterization on the nanoparticle surface, has CE offered very beneficial capabilities. Finally, the monitoring of interactions between nanomaterials and other (bio)molecules is easily performed by some kind of capillary electromigration technique. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  11. Comparison of comet 81P/Wild 2 dust with interplanetary dust from comets.

    Science.gov (United States)

    Ishii, Hope A; Bradley, John P; Dai, Zu Rong; Chi, Miaofang; Kearsley, Anton T; Burchell, Mark J; Browning, Nigel D; Molster, Frank

    2008-01-25

    The Stardust mission returned the first sample of a known outer solar system body, comet 81P/Wild 2, to Earth. The sample was expected to resemble chondritic porous interplanetary dust particles because many, and possibly all, such particles are derived from comets. Here, we report that the most abundant and most recognizable silicate materials in chondritic porous interplanetary dust particles appear to be absent from the returned sample, indicating that indigenous outer nebula material is probably rare in 81P/Wild 2. Instead, the sample resembles chondritic meteorites from the asteroid belt, composed mostly of inner solar nebula materials. This surprising finding emphasizes the petrogenetic continuum between comets and asteroids and elevates the astrophysical importance of stratospheric chondritic porous interplanetary dust particles as a precious source of the most cosmically primitive astromaterials.

  12. The asteroid-comet continuum from laboratory and space analyses of comet samples and micrometeorites

    Science.gov (United States)

    Engrand, Cecile; Duprat, Jean; Bardin, Noemie; Dartois, Emmanuel; Leroux, Hugues; Quirico, Eric; Benzerara, Karim; Rémusat, Laurent; Dobrică, Elena; Delauche, Lucie; Bradley, John; Ishii, Hope; Hilchenbach, Martin; COSIMA Team

    2015-08-01

    Comets are probably the best archives of the nascent solar system, 4.5 Gyr ago, and their compositions reveal crucial clues on the structure and dynamics of the early protoplanetary disk. Anhydrous minerals (olivine and pyroxene) have been identified in cometary dust for a few decades. Surprisingly, samples from comet Wild2 returned by the Stardust mission in 2006 also contain high temperature mineral assemblages like chondrules and refractory inclusions, which are typical components of primitive meteorites (carbonaceous chondrites - CCs). A few Stardust samples have also preserved some organic matter of comet Wild 2 that share some similarities with CCs. Interplanetary dust falling on Earth originate from comets and asteroids in proportions to be further constrained. These cosmic dust particles mostly show similarities with CCs, which in turn only represent a few percent of meteorites recovered on Earth. At least two (rare) families of cosmic dust particles have shown strong evidences for a cometary origin: the chondritic porous interplanetary dust particles (CP-IDPs) collected in the terrestrial stratosphere by NASA, and the ultracarbonaceous Antarctic Micrometeorites (UCAMMs) collected from polar snow and ice by French and Japanese teams. The Rosetta mission currently carries dust analyzers capable of measuring dust flux, sizes, physical properties and compositions of dust particles from the Jupiter family comet 67P/Churyumov-Gerasimenko (COSIMA, GIADA, MIDAS), as well as gas analyzers (ROSINA, PTOLEMY, COSAC). A growing number of evidences highlights the existence of a continuum between asteroids and comets, already in the early history of the solar system. We will present the implications of the analyses of samples in the laboratory and in space to a better understanding of the early protoplanetary disk.

  13. The persistent coma of Comet P/Schwassmann-Wachmann 1

    International Nuclear Information System (INIS)

    Jewitt, D.

    1990-01-01

    Time-series photometry of Comet P/Schwassmann-Wachmann 1 in both 1987 and 1988 shows that this comet is continually active despite its large heliocentric distance. The observed activity, upon which the famous outbursts of this comet are superposed, may be driven by the sublimation of crystalline water ice at the nucleus surface. A simple model which accounts for both the continuous activity and the sporadic outbursts is suggested. 34 refs

  14. Reservoirs for Comets: Compositional Differences Based on Infrared Observations

    Science.gov (United States)

    Disanti, Michael A.; Mumma, Michael J.

    Tracing measured compositions of comets to their origins continues to be of keen interest to cometary scientists and to dynamical modelers of Solar System formation and evolution. This requires building a taxonomy of comets from both present-day dynamical reservoirs: the Kuiper Belt (hereafter KB), sampled through observation of ecliptic comets (primarily Jupiter Family comets, or JFCs), and the Oort cloud (OC), represented observationally by the long-period comets and by Halley Family comets (HFCs). Because of their short orbital periods, JFCs are subjected to more frequent exposure to solar radiation compared with OC comets. The recent apparitions of the JFCs 9P/Tempel 1 and 73P/Schwassmann-Wachmann 3 permitted detailed observations of material issuing from below their surfaces—these comets added significantly to the compositional database on this dynamical class, which is under-represented in studies of cometary parent volatiles. This chapter reviews the latest techniques developed for analysis of high-resolution spectral observations from ˜2-5 μm, and compares measured abundances of native ices among comets. While no clear compositional delineation can be drawn along dynamical lines, interesting comparisons can be made. The sub-surface composition of comet 9P, as revealed by the Deep Impact ejecta, was similar to the majority of OC comets studied. Meanwhile, 73P was depleted in all native ices except HCN, similar to the disintegrated OC comet C/1999 S4 (LINEAR). These results suggest that 73P may have formed in the inner giant planets' region while 9P formed farther out or, alternatively, that both JFCs formed farther from the Sun but with 73P forming later in time.

  15. DNA electrophoresis through microlithographic arrays

    International Nuclear Information System (INIS)

    Sevick, E.M.; Williams, D.R.M.

    1996-01-01

    Electrophoresis is one of the most widely used techniques in biochemistry and genetics for size-separating charged molecular chains such as DNA or synthetic polyelectrolytes. The separation is achieved by driving the chains through a gel with an external electric field. As a result of the field and the obstacles that the medium provides, the chains have different mobilities and are physically separated after a given process time. The macroscopically observed mobility scales inversely with chain size: small molecules move through the medium quickly while larger molecules move more slowly. However, electrophoresis remains a tool that has yet to be optimised for most efficient size separation of polyelectrolytes, particularly large polyelectrolytes, e.g. DNA in excess of 30-50 kbp. Microlithographic arrays etched with an ordered pattern of obstacles provide an attractive alternative to gel media and provide wider avenues for size separation of polyelectrolytes and promote a better understanding of the separation process. Its advantages over gels are (1) the ordered array is durable and can be re-used, (2) the array morphology is ordered and can be standardized for specific separation, and (3) calibration with a marker polyelectrolyte is not required as the array is reproduced to high precision. Most importantly, the array geometry can be graduated along the chip so as to expand the size-dependent regime over larger chain lengths and postpone saturation. In order to predict the effect of obstacles upon the chain-length dependence in mobility and hence, size separation, we study the dynamics of single chains using theory and simulation. We present recent work describing: 1) the release kinetics of a single DNA molecule hooked around a point, frictionless obstacle and in both weak and strong field limits, 2) the mobility of a chain impinging upon point obstacles in an ordered array of obstacles, demonstrating the wide range of interactions possible between the chain and

  16. Comparison of non-electrophoresis grade with electrophoresis grade BIS in NIPAM polymer gel preparation

    Science.gov (United States)

    Khodadadi, Roghayeh; Khajeali, Azim; Farajollahi, Ali Reza; Hajalioghli, Parisa; Raeisi, Noorallah

    2015-01-01

    Introduction:The main objective of this study was to investigate the possibility of replacing electrophoresis cross-linker with non-electrophoresis N, N′-methylenebisacrylamide (BIS) in N-isopropyl acrylamide (NIPAM) polymer gel and its possible effect on dose response. Methods: NIPAM polymer gel was prepared from non-electrophoresis grade BIS and the relaxation rate (R2) was measured by MR imaging after exposing the gel to gamma radiation from Co-60 source. To compare the response of this gel with the one that contains electrophoresis grade BIS, two sets of NIPAM gel were prepared using electrophoresis and non-electrophoresis BIS and irradiated to different gamma doses. Results: It was found that the dose–response of NIPAM gel made from the non-electrophoresis grade BIS is coincident with that of electrophoresis grade BIS. Conclusion:Taken all, it can be concluded that the non-electrophoresis grade BIS not only is a suitable alternative for the electrophoresis grade BIS but also reduces the cost of gel due to its lower price. PMID:26457250

  17. Hormone assay

    International Nuclear Information System (INIS)

    Eisentraut, A.M.

    1977-01-01

    An improved radioimmunoassay is described for measuring total triiodothyronine or total thyroxine levels in a sample of serum containing free endogenous thyroid hormone and endogenous thyroid hormone bound to thyroid hormone binding protein. The thyroid hormone is released from the protein by adding hydrochloric acid to the serum. The pH of the separated thyroid hormone and thyroid hormone binding protein is raised in the absence of a blocking agent without interference from the endogenous protein. 125 I-labelled thyroid hormone and thyroid hormone antibodies are added to the mixture, allowing the labelled and unlabelled thyroid hormone and the thyroid hormone antibody to bind competitively. This results in free thyroid hormone being separated from antibody bound thyroid hormone and thus the unknown quantity of thyroid hormone may be determined. A thyroid hormone test assay kit is described for this radioimmunoassay. It provides a 'single tube' assay which does not require blocking agents for endogenous protein interference nor an external solid phase sorption step for the separation of bound and free hormone after the competitive binding step; it also requires a minimum number of manipulative steps. Examples of the assay are given to illustrate the reproducibility, linearity and specificity of the assay. (UK)

  18. Studying Short-Period Comets and Long-Period Comets Detected by WISE/NEOWISE

    Science.gov (United States)

    Kramer, Emily A.; Fernández, Yanga R.; Bauer, James M.; Stevenson, Rachel; Mainzer, Amy K.; Grav, Tommy; Masiero, Joseph; Walker, Russell G.; Lisse, Carey M.

    2014-11-01

    The Wide-field Infrared Survey Explorer (WISE) mission surveyed the sky in four infrared wavelength bands (3.4, 4.6, 12 and 22-micron) between January 2010 and February 2011 [1, 2]. During the mission, WISE serendipitously observed 160 comets, including 21 newly discovered objects. About 89 of the comets observed by WISE displayed a significant dust tail in the 12 and 22-micron (thermal emission) bands, showing a wide range of activity levels and dust morphology. Since the observed objects are a mix of both long-period comets (LPCs) and short-period comets (SPCs), differences in their activity can be used to better understand the thermal evolution that each of these populations has undergone. For the comets that displayed a significant dust tail, we have estimated the sizes and ages of the particles using dynamical models based on the Finson-Probstein method [3, 4]. For a selection of 40 comets, we have then compared these models to the data using a novel tail-fitting method that allows the best-fit model to be chosen analytically rather than subjectively. For comets that were observed multiple times by WISE, the dust tail particle properties were estimated separately, and then compared. We find that the dust tails of both LPCs and SPCs are primarily comprised of ~mm to cm sized particles, which were the result of emission that occurred several months to several years prior to the observations. The LPCs nearly all have strong dust emission close to the comet's perihelion distance, and the SPCs mostly have strong dust emission close to perihelion, but some have strong dust emission well before perihelion. Acknowledgments: This publication makes use of data products from (1) WISE, which is a joint project of UCLA and JPL/Caltech, funded by NASA; and (2) NEOWISE, which is a project of JPL/Caltech, funded by the Planetary Science Division of NASA. EK was supported by a NASA Earth and Space Sciences Fellowship. RS gratefully acknowledges support from the NASA

  19. Electrophoresis in space at zero gravity

    Science.gov (United States)

    Bier, M.; Snyder, R. S.

    1974-01-01

    Early planning for manufacturing operations in space include the use of electrophoresis for purification and separation of biological materials. Greatly simplified electrophoresis apparatus have been flown in the Apollo 14 and 16 missions to test the possibility of stable liquid systems in orbit. Additionally, isoelectric focusing and isotachophoresis are of particular interest as they offer very high resolution and have self-sharpening boundaries. The value of possible space electrophoresis is substantial. For example, present technology permits large fractionation of only a few of blood proteins many fractions, and separated cell populations are needed for research.

  20. Comet 81P/wild 2 under a microscope

    Science.gov (United States)

    Brownlee, D.; Tsou, P.; Aleon, J.; O'D, Alexander; Araki, T.; Bajt, S.; Baratta, G.A.; Bastien, R.; Bland, P.; Bleuet, P.; Borg, J.; Bradley, J.P.; Brearley, A.; Brenker, F.; Brennan, S.; Bridges, J.C.; Browning, N.D.; Brucato, J.R.; Bullock, E.; Burchell, M.J.; Busemann, H.; Butterworth, Anna L.; Chaussidon, M.; Cheuvront, A.; Chi, M.; Cintala, M.J.; Clark, B. C.; Clemett, S.J.; Cody, G.; Colangeli, L.; Cooper, G.; Cordier, P.; Daghlian, C.; Dai, Z.; D'Hendecourt, L.; Djouadi, Z.; Dominguez, G.; Duxbury, T.; Dworkin, J.P.; Ebel, D.S.; Economou, T.E.; Fakra, S.; Fairey, S.A.J.; Fallon, S.; Ferrini, G.; Ferroir, T.; Fleckenstein, H.; Floss, C.; Flynn, G.; Franchi, I.A.; Fries, M.; Gainsforth, Z.; Gallien, J.-P.; Genge, M.; Gilles, M.K.; Gillet, P.; Gilmour, J.; Glavin, D.P.; Gounelle, M.; Grady, M.M.; Graham, G.A.; Grant, P.G.; Green, S.F.; Grossemy, F.; Grossman, L.; Grossman, J.N.; Guan, Y.; Hagiya, K.; Harvey, R.; Heck, P.; Herzog, G.F.; Hoppe, P.; Horz, F.; Huth, J.; Hutcheon, I.D.; Ignatyev, K.; Ishii, H.; Ito, M.; Jacob, D.; Jacobsen, C.; Jacobsen, S.; Jones, S.; Joswiak, D.; Jurewicz, A.; Kearsley, A.T.; Keller, L.P.; Khodja, H.; Kilcoyne, A.L.D.; Kissel, J.; Krot, A.; Langenhorst, F.; Lanzirotti, A.; Le, L.; Leshin, L.A.; Leitner, J.; Lemelle, L.; Leroux, H.; Liu, M.-C.; Luening, K.; Lyon, I.; MacPherson, G.; Marcus, M.A.; Marhas, K.; Marty, B.; Matrajt, G.; McKeegan, K.; Meibom, A.; Mennella, V.; Messenger, K.; Messenger, S.; Mikouchi, T.; Mostefaoui, S.; Nakamura, T.; Nakano, T.; Newville, M.; Nittler, L.R.; Ohnishi, I.; Ohsumi, K.; Okudaira, K.; Papanastassiou, D.A.; Palma, R.; Palumbo, M.E.; Pepin, R.O.; Perkins, D.; Perronnet, M.; Pianetta, P.; Rao, W.; Rietmeijer, F.J.M.; Robert, F.; Rost, D.; Rotundi, A.; Ryan, R.; Sandford, S.A.; Schwandt, C.S.; See, T.H.; Schlutter, D.; Sheffield-Parker, J.; Simionovici, A.; Simon, S.; Sitnitsky, I.; Snead, C.J.; Spencer, M.K.; Stadermann, F.J.; Steele, A.; Stephan, T.

    2006-01-01

    The Stardust spacecraft collected thousands of particles from comet 81P/Wild 2 and returned them to Earth for laboratory study. The preliminary examination of these samples shows that the nonvolatile portion of the comet is an unequilibrated assortment of materials that have both presolar and solar system origin. The comet contains an abundance of silicate grains that are much larger than predictions of interstellar grain models, and many of these are high-temperature minerals that appear to have formed in the inner regions of the solar nebula. Their presence in a comet proves that the formation of the solar system included mixing on the grandest scales.

  1. X-rays from comets - a surprising discovery

    CERN Multimedia

    CERN. Geneva

    2000-01-01

    Comets are kilometre-size aggregates of ice and dust, which remained from the formation of the solar system. It was not obvious to expect X-ray emission from such objects. Nevertheless, when comet Hyakutake (C/1996 B2) was observed with the ROSAT X-ray satellite during its close approach to Earth in March 1996, bright X-ray emission from this comet was discovered. This finding triggered a search in archival ROSAT data for comets, which might have accidentally crossed the field of view during observations of unrelated targets. To increase the surprise even more, X-ray emission was detected from four additional comets, which were optically 300 to 30 000 times fainter than Hyakutake. For one of them, comet Arai (C/1991 A2), X-ray emission was even found in data which were taken six weeks before the comet was optically discovered. These findings showed that comets represent a new class of celestial X-ray sources. The subsequent detection of X-ray emission from several other comets in dedicated observations confir...

  2. Polyacrylamide gel electrophoresis of RNA.

    Science.gov (United States)

    Rio, Donald C; Ares, Manuel; Hannon, Gregory J; Nilsen, Timothy W

    2010-06-01

    Perhaps the most important and certainly the most often used technique in RNA analysis is gel electrophoresis. This technique is generally applicable for RNA detection, quantification, purification by size, and quality assessment. Because RNAs are negatively charged, they migrate toward the anode in the presence of electric current. The gel acts as a sieve to selectively impede the migration of the RNA in proportion to its mass, given that its mass is generally proportional to its charge. Because mass is approximately related to chain length, the length of an RNA is more generally determined by its migration. In addition, topology (i.e., circularity) can affect migration, making RNAs appear longer on the gel than they actually are. Gels are used in a wide variety of techniques, including Northern blotting, primer extension, footprinting, and analyzing processing reactions. They are invaluable as preparative and fractionating tools. There are two common types of gel: polyacrylamide and agarose. For most applications, denaturing acrylamide gels are most appropriate. These gels are extremely versatile and can resolve RNAs from ~600 to RNA-protein complexes, native gels are appropriate. The only disadvantage to acrylamide gels is that they are not suitable for analyzing large RNAs (> or =600 nt); for such applications, agarose gels are preferred. This protocol describes how to prepare, load, and run polyacrylamide gels for RNA analysis.

  3. Evaluation of wheat by polyacrylamide gel electrophoresis

    African Journals Online (AJOL)

    PRECIOUS

    2010-01-11

    04, Inqulab-91 and Rawal-. 87 were evaluated for analysis of variability in seed storage proteins by using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Electrophorogram for each variety were scored ...

  4. Characterization of asphaltenes by nonaqueous capillary electrophoresis

    NARCIS (Netherlands)

    Kok, W.T.; Tüdös, A.J.; Grutters, M.; Shepherd, A.G.

    2011-01-01

    Nonaqueous capillary electrophoresis was used for the separation and characterization of asphaltene samples from different sources. For the separation medium (background electrolyte), mixtures of tetrahydrofuran and a high-permittivity organic solvent could be used. The best results were obtained

  5. High speed single nucleotide polymorphism typing of a hereditary haemochromatosis mutation with capillary array electrophoresis microplates.

    Science.gov (United States)

    Medintz, I; Wong, W W; Sensabaugh, G; Mathies, R A

    2000-07-01

    A single nucleotide polymorphism (SNP) typing assay is developed and evaluated on a microfabricated capillary array electrophoresis system. Using fluorescently labeled allele-specific primers, the S65C (193A-->T) substitution associated with hereditary haemochromatosis in the HFE gene is genotyped. The covalently labeled polymerase chain reaction (PCR) products are separated on a microfabricated radial capillary array electrophoresis microplate using nondenaturing gel media in under two minutes. Detection is accomplished with a laser-excited rotary confocal scanner. The Rox-labeled A-allele specific amplicon (211 bp) is differentiated from the R110-labeled T-allele specific amplicon (201 bp) by both size and color. This study demonstrates the feasibility of using allele-specific PCR with covalently labeled primers for high speed fluorescent SNP typing on microfabricated radial capillary array electrophoresis microplates.

  6. Conducting Polymer Electrodes for Gel Electrophoresis

    OpenAIRE

    Bengtsson, Katarina; Nilsson, Sara; Robinson, Nathaniel D

    2014-01-01

    In nearly all cases, electrophoresis in gels is driven via the electrolysis of water at the electrodes, where the process consumes water and produces electrochemical by-products. We have previously demonstrated that p-conjugated polymers such as poly(3,4-ethylenedioxythiophene) (PEDOT) can be placed between traditional metal electrodes and an electrolyte to mitigate electrolysis in liquid (capillary electroosmosis/electrophoresis) systems. In this report, we extend our previous result to gel ...

  7. DNA-electrophoresis of single cells - a method to screen for irradiated foodstuffs

    International Nuclear Information System (INIS)

    Leffke, A.; Helle, N.; Boegl, K.W.; Schreiber, G.A.

    1993-01-01

    Microelectrophoresis of single cells can be used to detect γ-irradiation over a wide dose range and for a variety of products. It is a simple and rapid test for DNA damages and can be used for screening. The method was tested on cell suspensions of bone marrow and muscle cells from frozen chicken legs, chicken heart, turkey liver, beef and pork irradiated with doses up to 3 kGy. Cell suspensions were prepared by incubation of tissues in EDTA-SDS-buffer at pH 8. Single cell electrophoresis was performed in 0.75% agarose gel. DNA was visualised by silver staining. In unirradiated samples no or only a small amount of DNA penetrated the cell membranes. Cells of irradiated samples appeared like a ''comet'' due to to migration of DNA-fragments out of cell. (orig.)

  8. Comet and meteorite traditions of Aboriginal Australians

    Science.gov (United States)

    Hamacher, Duane W.

    2014-06-01

    This research contributes to the disciplines of cultural astronomy (the academic study of how past and present cultures understand and utilise celestial objects and phenomena) and geomythology (the study of geological events and the formation of geological features described in oral traditions). Of the hundreds of distinct Aboriginal cultures of Australia, many have oral traditions rich in descriptions and explanations of comets, meteors, meteorites, airbursts, impact events, and impact craters. These views generally attribute these phenomena to spirits, death, and bad omens. There are also many traditions that describe the formation of meteorite craters as well as impact events that are not known to Western science.

  9. Workshop on Observations of Recent Comets (1990)

    Science.gov (United States)

    Huebner, Walter F. (Editor); Wehinger, Peter A. (Editor); Rahe, Jurgen (Editor); Konno, Ichishiro (Editor)

    1990-01-01

    Potential interpretations are presented for observations of four comets: Brorsen-Metcalf (1989o), Okazaki-Levy-Rudenko (1989r), Aarseth-Brewington (1989a1), and Austin (1989o1). The relationship of minor species with each other and possible parents as well as with dust are being pursued in a number of investigations. Of particular interest are the abundance ratios of CH4 to CO and NH3 to N2. The need for closer collaboration betwen observing teams and modelers is examined. The need for dust size distribution as a function of cometocentric distance to be analyzed in closer collaboration between observers and modelers is discussed.

  10. Mission to a comet that could save earth

    CERN Multimedia

    Utton, T

    2003-01-01

    Scientists are going to attempt to land a probe on the comet Wirtanen. The GBP640million unmanned craft will travel four billion miles before catching up with the comet Wirtanen and launching a robotic lander called Rosetta, on to its surface (1/2 page).

  11. Dormant Comets in the Near-Earth Asteroid Population

    NARCIS (Netherlands)

    Mommert, Michael; Harris, Alan W.; Mueller, Michael; Hora, Joseph L.; Trilling, David E.; Knight, Matthew; Bottke, William F.; Thomas, Cristina; Delbo', Marco; Emery, Josh P.; Fazio, Giovanni; Smith, Howard A.

    2015-01-01

    The population of near-Earth objects comprises active comets and asteroids, covering a wide range of dynamical parameters and physical properties. Dormant (or extinct) comets, masquerading as asteroids, have long been suspected of supplementing the near-Earth asteroid (NEA) population. We present a

  12. The Formation Age of Comets: Predicted Physical and Chemical Trends

    Science.gov (United States)

    Nuth, J. A., III; Hill, H. G. M.

    2000-01-01

    The chemical composition of a comet has always been considered to be a function of where it formed in the nebula. We suggest that the most important factor in determining a comet's chemistry might actually be when it formed. Specific predictions are presented.

  13. Direct Detection of Extra-Solar Comets is Possible

    OpenAIRE

    Jura, M.

    2005-01-01

    The dust tails of comets similar to Hale-Bopp can scatter as much optical light as does the Earth. Space-based observatories such as the Terrestrial Planet Finder or Darwin that will detect extra-solar terrestrial planets also will be able to detect extra-solar comets.

  14. Ensemble Properties of Comets in the Sloan Digital Sky Survey

    Energy Technology Data Exchange (ETDEWEB)

    Solontoi, Michael; /Adler Planetarium, Chicago; Ivezic, Zeljko; /Washington U., Seattle, Astron. Dept.; Juric, Mario; /Harvard Coll. Observ.; Becker, Andrew C.; /Washington U., Seattle, Astron. Dept.; Jones, Lynne; /Washington U., Seattle, Astron. Dept.; West, Andrew A.; /Boston U.; Kent, Steve; /Fermilab; Lupton, Robert H.; /Princeton U. Observ.; Claire, Mark; /Washington U., Seattle, Astron. Dept.; Knapp, Gillian R.; /Princeton U. Observ.; Quinn, Tom; /Washington U., Seattle, Astron. Dept. /Princeton U. Observ.

    2012-02-01

    We present the ensemble properties of 31 comets (27 resolved and 4 unresolved) observed by the Sloan Digital Sky Survey (SDSS). This sample of comets represents about 1 comet per 10 million SDSS photometric objects. Five-band (u, g, r, i, z) photometry is used to determine the comets colors, sizes, surface brightness profiles, and rates of dust production in terms of the Afp formalism. We find that the cumulative luminosity function for the Jupiter Family Comets in our sample is well fit by a power law of the form N(comets. The resolved comets show an extremely narrow distribution of colors (0.57 {+-} 0.05 in g - r for example), which are statistically indistinguishable from that of the Jupiter Trojans. Further, there is no evidence of correlation between color and physical, dynamical, or observational parameters for the observed comets.

  15. Radioreceptor assays

    International Nuclear Information System (INIS)

    Lapka, R.

    1985-01-01

    Radioreceptor assay (RRA) is an analytical method using the specific interaction of some pharmaceuticals and endogenic substances (ligands) with specific receptors present in certin tissues of living organisms. RRA uses the principle of isotope dilution. The method is described in detail of the preparation of receptors, samples and radioligands, conditions of incubation, the separation of free and bound radioligand, and the mathematical evaluation of RRA. The sensitivity of RRA is measured in units to tens of pg. The specificity of RRA relates to a group of substances with similar pharmacological effect. RRA may be used for identifying neuroleptics, antidepressants, anxiolytics, ergot alkaloids, beta blockers, anticholinergic drugs, certain hormones and neuropeptides. (M.D.)

  16. The exploration of Halley's comet - An example of international cooperation

    Science.gov (United States)

    Rahe, Jurgen H.; Newburn, Ray L., Jr.

    1987-01-01

    The history of international cooperation in studies of comets started with observations in 1577 and 1680, when Tycho Brahe and Newton, respectively, collected position measurements made in different countries to determine the paths of the comets observed. In the fall of 1979, a worldwide Comet Halley watch was proposed. As a result of international cooperation, Comet Halley was explored during its recent appearance from the ground, earth orbit, Venus orbit, interplanetary space, and from within the comet itself. The various activities in space were coordinated by the ESA, the USSR Intercosmos, the Japanese ISAS, and NASA, through the Inter-Agency Consultative Group. The activities of the ground-based observers were coordinated by the International Halley Watch.

  17. THE PHOTODISSOCIATION OF FORMALDEHYDE IN COMETS

    Energy Technology Data Exchange (ETDEWEB)

    Feldman, Paul D., E-mail: pfeldman@jhu.edu [Department of Physics and Astronomy, The Johns Hopkins University, 3400 N. Charles Street, Baltimore, MD 21218 (United States)

    2015-10-20

    Observations of comets in the 905–1180 Å spectral band made with the Far Ultraviolet Spectroscopic Explorer in 2001 and 2004 show unusual features in the fluorescent emissions of CO and H{sub 2}. These include emission from a non-thermal high-J rotational population of CO and solar Lyα induced fluorescence from excited vibrational levels of H{sub 2}, both of which are attributed to the photodissociation of formaldehyde. In this paper we model the large number of observed H{sub 2} lines and demonstrate the dependence of the pumping on the heliocentric velocity of the comet and the solar line profiles. We also derive the rotational and vibrational populations of H{sub 2} and show that they are consistent with the results of laboratory studies of the photodissociation of H{sub 2}CO. In addition to the principal series of H i and O i, the residual spectrum is found to consist mainly of the Rydberg series of C i multiplets from which we derive the mean carbon column abundance in the coma. Fluorescent emissions from N i and N{sub 2} are also searched for.

  18. Rapid monitoring of autolysis process of proteases by capillary electrophoresis.

    Science.gov (United States)

    Chen, Xiu-Lan; Shun, Cai-Yun; Zhang, Yu-Zhong; Gao, Pei-Ji

    2003-10-01

    A protease, MCP-01, produced by a deep-sea psychrotrophic strain of Pseudoaltermonas sp. SM9913 was purified and its autolysis reaction at 20 degrees C-50 degrees C was monitored by capillary electrophoresis. Capillary electrophoresis provides a rapid assay because the degree and state of autolysis of protease MCP-01 could be observed within 6 min. The autolysis rate increased as the temperature rose in the tested range. After 30 min incubation at 30 degrees C, 77% of MCP-01 autolyzed into peptides. However, its activity for the hydrolysis of casein was reduced by only 4%. The rate of loss of activity of MCP-01 was thus slower than that of autolysis of MCP-01 at 30 degrees C. Similar results were obtained when MCP-01 was incubated at 20 degrees C, 40 degrees C and 50 degrees C. Large peptides produced by autolysis of MCP-01 therefore still have catalytic activity. When these large peptides autolyzed further into smaller peptides, the enzyme conformation that retained its catalytic activity was destroyed and activity was lost.

  19. Microchip analysis of lithium in blood using moving boundary electrophoresis and zone electrophoresis

    NARCIS (Netherlands)

    Vrouwe, E.X.; Lüttge, Regina; Olthuis, Wouter; van den Berg, Albert

    The determination of inorganic cations in blood plasma is demonstrated using a combination of moving boundary electrophoresis (MBE) and zone electrophoresis. The sample loading performed under MBE conditions is studied with the focus on the quantitative analysis of lithium. A concentration

  20. Comet Hunters: A Citizen Science Project to Search for Comets in the Main Asteroid Belt

    Science.gov (United States)

    Hsieh, Henry H.; Schwamb, Megan Elizabeth; Zhang, Zhi-Wei; Chen, Ying-Tung; Wang, Shiang-Yu; Lintott, Chris

    2016-10-01

    Fully automated detection of comets in wide-field surveys remains a challenge, as even highly successful comet-finding surveys like Pan-STARRS rely on a combination of both automated flagging algorithms and vetting by human eyes. To take advantage of the long-noted superiority of the human eye over computer algorithms in certain types of pattern recognition, particularly when dealing with a range of target morphologies of interest, we have created a citizen science website with the aim of allowing the general public to aid in the search for active asteroids, which are objects that occupy dynamically asteroidal orbits yet exhibit comet-like dust emission due to sublimation, impact disruption, rotational destabilization, or other effects. Located at comethunters.org, the Comet Hunters website was built using the Zooniverse Project Builder (https://www.zooniverse.org/lab), and displays images of known asteroids obtained either from archival data obtained between 1999 and 2014 by the Suprime-Cam wide-field imager mounted on the 8-m Subaru telescope on Mauna Kea in Hawaii, or more contemporary data obtained by the Hyper Suprime-Cam (HSC) wide-field imager also on the Subaru Telescope as part of the ongoing HSC Subaru Strategic Program (SSP) survey. By using observations from such a large-aperture telescope, most of which have never been searched for solar system objects, much less cometary ones, we expect that volunteers should be able to make genuinely scientifically significant discoveries, and also provide valuable insights into the potential and challenges of searching for comets in the LSST era. To date, over 13,000 registered volunteers have contributed 350,000 classifications. We will discuss the design and construction of the Comet Hunters website, and also discuss early results from the project.This work uses data generated via the Zooniverse.org platform, development of which was supported by a Global Impact Award from Google, and by the Alfred P. Sloan

  1. Analysis of RNA by capillary electrophoresis.

    Science.gov (United States)

    Skeidsvoll, J; Ueland, P M

    1996-09-01

    Analytical parameters known to be important for the separation of DNA by capillary electrophoresis, including gel polymer concentration, electrical field strength and temperature, were investigated and optimized for the analysis of RNA molecules from 100 to 2000 bases. Denaturation, essential to obtain uniform and identifiable peaks, was accomplished by heating the sample in 80% formamide prior to electrophoresis and the presence of 2-8 M urea in the electrophoresis buffer. Efficient separations were obtained over a wide range of electrical field strengths and temperatures using the gel polymer hydroxypropylmethylcellulose (HPMC) as separation matrix. Low HPMC concentrations (RNA (> 1000 bases) whereas higher HPMC concentrations were required for optimal separation of low molecular mass RNA. An optimized system was applicable for the separation of the predominating RNA populations (small RNA of 60-300 bases (as a group of unseparated peaks), 18S and 28S rRNA) in total RNA from a human glioma cell line. This is the first systematic investigation of electrophoresis of higher molecular mass RNA in capillaries, and motivates further studies to transfer electrophoresis of RNA to the capillary format.

  2. Glycosaminoglycan blotting and detection after electrophoresis separation.

    Science.gov (United States)

    Volpi, Nicola; Maccari, Francesca

    2015-01-01

    Separation of glycosaminoglycans (GAGs) by electrophoresis and their characterization to the microgram level are integral parts of biochemical research. Their blotting on membranes after electrophoresis offers the advantage to perform further analysis on single separated species such as identification with antibodies and/or recovery of single band. A method for the blotting and immobilizing of several nonsulfated and sulfated complex GAGs on membranes made hydrophilic and positively charged by cationic detergent after their separation by conventional agarose-gel electrophoresis is illustrated. This approach to the study of these complex macromolecules utilizes the capacity of agarose-gel electrophoresis to separate single species of polysaccharides from mixtures and the membrane technology for further preparative and analytical uses. Nitrocellulose membranes are derivatized with the cationic detergent cetylpyridinium chloride (CPC) and mixtures of GAGs are capillary blotted after their separation in agarose-gel electrophoresis. Single purified species of variously sulfated polysaccharides are transferred on derivatized membranes with an efficiency of 100 % and stained with alcian blue (irreversible staining) and toluidine blue (reversible staining). This enables a lower amount limit of detection of 0.1 μg. Nonsulfated polyanions, for example hyaluronic acid (HA), may also be transferred to membranes with a limit of detection of approximately 0.1-0.5 μg after irreversible or reversible staining. The membranes may be stained with reversible staining and the same lanes used for immunological detection or other applications.

  3. Detection of miRNA in cell cultures by using microchip electrophoresis with a fluorescence-labeled riboprobe.

    Science.gov (United States)

    Yamamura, Shohei; Yatsushiro, Shouki; Yamaguchi, Yuka; Abe, Kaori; Shinohara, Yasuo; Kataoka, Masatoshi

    2012-01-01

    The analysis of a microRNA (miRNA), miR-222 isolated from the PC12 cell line, was performed by use of the ribonuclease (RNase) protection assay, cyanine 5 (Cy5)-labeled miR-222 riboprobe, and a Hitachi SV1210 microchip electrophoresis system, which can be used to evaluate the integrity of total RNA. The fluorescence intensity corresponding to the protected RNA fragment increased in a dose-dependent manner with respect to the complementary-strand RNA. More highly sensitive detection of miRNA by microchip electrophoresis than by conventional method using fluorescence-labeled riboprobe could be obtained in 180 s. An obvious increase in miR-222 expression induced by nerve growth factor in PC12 cells could be observed. These results clearly indicate the potential of microchip electrophoresis for the analysis of miRNA using RNase protection assay with a fluorescence-labeled riboprobe.

  4. Perspectives on Comets, Comet-like Asteroids, and Their Predisposition to Provide an Environment That Is Friendly to Life.

    Science.gov (United States)

    Bosiek, Katharina; Hausmann, Michael; Hildenbrand, Georg

    2016-04-01

    In recent years, studies have shown that there are many similarities between comets and asteroids. In some cases, it cannot even be determined to which of these groups an object belongs. This is especially true for objects found beyond the main asteroid belt. Because of the lack of comet fragments, more progress has been made concerning the chemical composition of asteroids. In particular, the SMASSII classification establishes a link between the reflecting spectra and chemical composition of asteroids and meteorites. To find clues for the chemical structure of comets, the parameters of all known asteroids of the SMASSII classification were compared to those of comet groups like the Encke-type comets, the Jupiter-family comets, and the Halley-type comets, as well as comet-like objects like the damocloids and the centaurs. Fifty-six SMASSII objects similar to comets were found and are categorized as comet-like asteroids in this work. Aside from the chemistry, it is assumed that the available energy on these celestial bodies plays an important role concerning habitability. For the determination of the available energy, the effective temperature was calculated. Additionally, the size of these objects was considered in order to evaluate the possibility of a liquid water core, which provides an environment that is more likely to support processes necessary to create the building blocks of life. Further study of such objects could be notable for the period of the Late Heavy Bombardment and could therefore provide important implications for our understanding of the inner workings of the prebiotic evolution within the Solar System since the beginning.

  5. P126-T A Rapid and Sensitive Method for RNA Quality Determination on Capillary Electrophoresis Systems

    OpenAIRE

    Shah, A. B.; Karudapuram, S.; Joe, L. K.; Hauser, J.; Huynh, A. T.; Pothini, S.; Samsani, S.; Vennemeyer, E. S.; Briggs, J.; Wenz, M.; Waldron, C.; Buckelew, A.; Carver, C.; Bass, S.; Baumgartner, P.

    2007-01-01

    RNA quality is directly correlated to the success of various applications, such as microarray or real time qPCR-based gene expression analyses, cDNA library construction, Northern analyses, and RNAse protection assays, which utilize RNA samples from various organisms, tissues, cell lines, and precious biological samples. We present a novel method for examining RNA integrity and purity using capillary electrophoresis that is more cost-effective and scalable than current standard methods. This ...

  6. Preparative electrophoresis of industrial fission product solutions

    International Nuclear Information System (INIS)

    Tret, Joel

    1971-07-01

    The aim of this work is to contribute to the development of the continuous electrophoresis technique while studying its application in the preparative electrophoresis of industrial fission product solutions. The apparatus described is original. It was built for the purposes of the investigation and proved very reliable in operation. The experimental conditions necessary to maintain and supervise the apparatus in a state of equilibrium are examined in detail; their stability is an important factor, indispensable to the correct performance of an experiment. By subjecting an industrial solution of fission products to preparative electrophoresis it is possible, according to the experimental conditions, to prepare carrier-free radioelements of radiochemical purity (from 5 to 7 radioelements): 137 Cs, 90 Sr, 141+144 Ce, 91 Y, 95 Nb, 95 Zr, 103+106 Ru. (author) [fr

  7. Denaturation and electrophoresis of RNA with formaldehyde.

    Science.gov (United States)

    Rio, Donald C

    2015-02-02

    Electrophoretic size fractionation can be used to denature and separate large mRNA molecules (0.5-10 kb) on formaldehyde-containing agarose gels. Formaldehyde contains a carbonyl group that reacts to form Schiff bases with the imino or amino groups of guanine, adenine, and cytosine. These covalent adducts prevent normal base pairing and maintain the RNA in a denatured state. Because these adducts are unstable, formaldehyde must be present in the gel to maintain the RNA in the denatured state. This protocol describes the preparation of an agarose gel with formaldehyde and its setup in a horizontal electrophoresis apparatus. RNA samples are prepared and denatured in a solution of formamide and formaldehyde and, with 0.5- to 10-kb size markers, subjected to electrophoresis through the gel. Following electrophoresis, the gel is stained to visualize RNA markers or rRNA using one of several different types of stains. © 2015 Cold Spring Harbor Laboratory Press.

  8. COMET SHOWERS ARE NOT INDUCED BY INTERSTELLAR CLOUDS

    Energy Technology Data Exchange (ETDEWEB)

    Morris, D.E.

    1985-11-01

    Encounters with interstellar clouds (IC) have been proposed by Rampino and Stothers as a cause of quasi-periodic intense comet showers leading to earth impacts, in order to explain the periodicity in marine mass extinctions found by Raup and Sepkoski. The model was described further, criticized and defended. The debate has centered on the question of whether the scale height of the clouds is small enough (in comparison to the amplitude of the oscillation of the solar system about the plane of the Galaxy) to produce a modulation in the rate of encounters. We wish to point out another serious, we believe fatal, defect in this model - the tidal fields of ICs are not strong enough to produce intense comet showers leading to earth impacts by bringing comets of the postulated inner Oort cloud into earth crossing orbits, except possibly during very rare encounters with very dense clouds. We will show that encounters with abundant clouds of low density cannot produce comet showers; cloud density N > 10{sup 3} atoms cm{sup -3} is needed to produce an intense comet shower leading to earth impacts. Furthermore, the tidal field of a dense cloud during a distant encounter is too weak to produce such showers. As a consequence, comet showers induced by ICs will be far less frequent than showers caused by passing stars. This conclusion is independent of assumptions about the radial distribution of comets in the inner Oort cloud.

  9. 21 CFR 864.7455 - Fetal hemoglobin assay.

    Science.gov (United States)

    2010-04-01

    ... hemoglobin polypeptide chains). The hemoglobin determination may be made by methods such as electrophoresis... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Fetal hemoglobin assay. 864.7455 Section 864.7455...) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Hematology Kits and Packages § 864.7455 Fetal hemoglobin...

  10. Outbursting Comet P/2010 V1 (Ikeya-Murakami): A Miniature Comet Holmes

    Science.gov (United States)

    Ishiguro, Masateru; Jewitt, David; Hanayama, Hidekazu; Usui, Fumihiko; Sekiguchi, Tomohiko; Yanagisawa, Kenshi; Kuroda, Daisuke; Yoshida, Michitoshi; Ohta, Kouji; Kawai, Nobuyuki; Miyaji, Takeshi; Fukushima, Hideo; Watanabe, Jun-ichi

    2014-05-01

    The short-period comet P/2010 V1 (Ikeya-Murakami, hereafter "V1") was discovered visually by two amateur astronomers. The appearance of the comet was peculiar, consisting of an envelope, a spherical coma near the nucleus and a tail extending in the anti-solar direction. We investigated the brightness and the morphological development of the comet by taking optical images with ground-based telescopes. Our observations show that V1 experienced a large-scale explosion between UT 2010 October 31 and November 3. The color of the comet was consistent with the Sun (g' - R C = 0.61 ± 0.20, R C - I C = 0.20 ± 0.20, and B - R C = 0.93 ± 0.25), suggesting that dust particles were responsible for the brightening. We used a dynamical model to understand the peculiar morphology, and found that the envelope consisted of small grains (0.3-1 μm) expanding at a maximum speed of 500 ± 40 m s-1, while the tail and coma were composed of a wider range of dust particle sizes (0.4-570 μm) and expansion speeds 7-390 m s-1. The total mass of ejecta is ~5 × 108 kg and kinetic energy ~5 × 1012 J. These values are much smaller than in the historic outburst of 17P/Holmes in 2007, but the energy per unit mass (1 × 104 J kg-1) is comparable. The energy per unit mass is about 10% of the energy released during the crystallization of amorphous water ice suggesting that crystallization of buried amorphous ice can supply the mass and energy of the outburst ejecta.

  11. Outbursting comet P/2010 V1 (Ikeya-Murakami): A miniature comet Holmes

    Energy Technology Data Exchange (ETDEWEB)

    Ishiguro, Masateru [Department of Physics and Astronomy, Seoul National University, Gwanak, Seoul 151-742 (Korea, Republic of); Jewitt, David [Department of Earth, Planetary and Space Sciences, University of California at Los Angeles, 595 Charles Young Drive East, Los Angeles, CA 90095-1567 (United States); Hanayama, Hidekazu; Miyaji, Takeshi; Fukushima, Hideo; Watanabe, Jun-ichi [Ishigakijima Astronomical Observatory, National Astronomical Observatory of Japan, Ishigaki, Okinawa 907-0024 (Japan); Usui, Fumihiko [Department of Astronomy, Graduate School of Science, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033 (Japan); Sekiguchi, Tomohiko [Department of Teacher Training, Hokkaido University of Education, 9 Hokumon, Asahikawa 070-8621 (Japan); Yanagisawa, Kenshi; Kuroda, Daisuke [Okayama Astrophysical Observatory, National Astronomical Observatory of Japan, Asaguchi, Okayama 719-0232 (Japan); Yoshida, Michitoshi [Hiroshima Astrophysical Science Center, Hiroshima University, 1-3-1 Kagamiyama, Higashi-Hiroshima, Hiroshima 739-8526 (Japan); Ohta, Kouji [Department of Astronomy, Kyoto University, Kyoto 606-8502 (Japan); Kawai, Nobuyuki [Department of Physics, Tokyo Institute of Technology, 2-12-1 Ookayama, Meguro-ku, Tokyo 152-8551 (Japan)

    2014-05-20

    The short-period comet P/2010 V1 (Ikeya-Murakami, hereafter {sup V}1{sup )} was discovered visually by two amateur astronomers. The appearance of the comet was peculiar, consisting of an envelope, a spherical coma near the nucleus and a tail extending in the anti-solar direction. We investigated the brightness and the morphological development of the comet by taking optical images with ground-based telescopes. Our observations show that V1 experienced a large-scale explosion between UT 2010 October 31 and November 3. The color of the comet was consistent with the Sun (g' – R {sub C} = 0.61 ± 0.20, R {sub C} – I {sub C} = 0.20 ± 0.20, and B – R {sub C} = 0.93 ± 0.25), suggesting that dust particles were responsible for the brightening. We used a dynamical model to understand the peculiar morphology, and found that the envelope consisted of small grains (0.3-1 μm) expanding at a maximum speed of 500 ± 40 m s{sup –1}, while the tail and coma were composed of a wider range of dust particle sizes (0.4-570 μm) and expansion speeds 7-390 m s{sup –1}. The total mass of ejecta is ∼5 × 10{sup 8} kg and kinetic energy ∼5 × 10{sup 12} J. These values are much smaller than in the historic outburst of 17P/Holmes in 2007, but the energy per unit mass (1 × 10{sup 4} J kg{sup –1}) is comparable. The energy per unit mass is about 10% of the energy released during the crystallization of amorphous water ice suggesting that crystallization of buried amorphous ice can supply the mass and energy of the outburst ejecta.

  12. Numerical models of Oort Cloud formation and comet delivery

    Science.gov (United States)

    Kaib, Nathan A.

    I use a newly designed numerical algorithm to simulate the dynamics of the Oort Cloud. The processes I model are the formation of the cloud, the current delivery of comets to the planetary region, and long-period comet production during comet showers. Concerning the cloud's formation, I find that the Sun's birth environment dramatically affects the structure of the inner Oort Cloud as well as the amount of material trapped in this region. In addition, the structure of this reservoir is also sensitive to the Sun's orbital history in the Milky Way. This raises the possibility that constraining our inner Oort Cloud's properties can constrain the Sun's dynamical history. In this regard, I use my simulations of comet delivery to better understand what the population of comets passing through the planetary region can tell us about the inner Oort Cloud. I find that the inner Oort Cloud (rather than the scattered disk) dominates the production of planet-crossing TNOs with perihelia beyond 15 AU and semimajor axes greater than a few hundred AU. My results indicate that two objects representing this population (2000 00 67 and 2006 SQ 372 ) have already been detected, and the detection of many analogous objects can constrain the inner Oort Cloud. In addition, these simulations of comet delivery also demonstrate that, contrary to previous understanding, the inner Oort Cloud is a significant and perhaps the dominant source of known long-period comets. This result can be used to place the first observationally motivated upper limit on the inner Oort Cloud's population. Finally, with this maximum population value, I use my comet shower simulations to show that comet showers are unlikely to be responsible for more than one minor extinction event since the Cambrian Explosion.

  13. Comet LINEAR C/1999 S4 - an absolutely well-behaved comet before breakup

    Science.gov (United States)

    Peschke, S. B.; Lisse, C. M.; Fernandez, Y. R.; Ressler, M.; Stickel, M.; Kaminski, C.; Golish, B.

    2000-10-01

    We present results from infrared imaging of comet LINEAR C/1999 S4 on June 17 - 19, 2000 (pre-breakup), using the near-IR camera NSFCAM and the mid-IR camera MIRLIN at the 3m NASA/IRTF. Images and multi-wavelength spectroscopy were obtained in the zJHK'L'MNQ bands, and were used to create a 1.0 - 25 μ m SED of the comet's dust and nucleus. The coma's contribution at each wavelength was modeled using spatial fitting (Fernandez 1999, PhD thesis; Lisse et al. 1999, Icarus 140, 189). The resulting comatic and nuclear SEDs were then modeled using modified Mie theory (Lisse et al. 1998, ApJ 496, 971) and the standard nuclear thermal models (Lebofsky and Spencer 1989, Asteroids II, 128), respectively. We report the resulting dust PSD, mass loss rate, and albedo, as well as the nuclear radius, and we compare these results to those obtained by others from optical data both before and after the comet's breakup in late July 2000.

  14. Application of Microchip Electrophoresis for Clinical Tests

    Science.gov (United States)

    Yatsushiro, Shouki; Kataoka, Masatoshi

    Microchip electrophoresis has recently attracted much attention in the field of nuclear acid analysis due to its high efficiency, ease of operation, low consumption of samples and reagents, and relatively low costs. In addition, the analysis has expanded to an analytical field like not only the analysis of DNA but also the analysis of RNA, the protein, the sugar chain, and the cellular function, etc. In this report, we showed that high-performance monitoring systems for human blood glucose levels and α-amylase activity in human plasma using microchip electrophoresis.

  15. Undergraduate physics laboratory: Electrophoresis in chromatography paper

    Science.gov (United States)

    Hyde, Alexander; Batishchev, Oleg

    2015-12-01

    An experiment studying the physical principles of electrophoresis in liquids was developed for an undergraduate laboratory. We have improved upon the standard agarose gel electrophoresis experimental regime with a straightforward and cost-effective procedure, in which drops of widely available black food coloring were separated by electric field into their dye components on strips of chromatography paper soaked in a baking soda/water solution. Terminal velocities of seven student-safe dyes were measured as a function of the electric potential applied along the strips. The molecular mobility was introduced and calculated by analyzing data for a single dye. Sources of systematic and random errors were investigated.

  16. Contact Charge Electrophoresis: Fundamentals and Microfluidic Applications.

    Science.gov (United States)

    Bishop, Kyle J M; Drews, Aaron M; Cartier, Charles A; Pandey, Shashank; Dou, Yong

    2018-01-31

    Contact charge electrophoresis (CCEP) uses steady electric fields to drive the oscillatory motion of conductive particles and droplets between two or more electrodes. In contrast to traditional forms of electrophoresis and dielectrophoresis, CCEP allows for rapid and sustained particle motions driven by low-power dc voltages. These attributes make CCEP a promising mechanism for powering active components for mobile microfluidic technologies. This Feature Article describes our current understanding of CCEP as well as recent strategies to harness it for applications in microfluidics and beyond.

  17. Mars Reconnaissance Orbiter Navigation Strategy for the Comet Siding Spring Encounter

    Science.gov (United States)

    Menon, Premkumar R.; Wagner, Sean V.; Martin-Mur, Tomas J.; Jefferson, David C.; Ardalan, Shadan M.; Chung, Min-Kun J.; Lee, Kyong J.; Schulze, William B.

    2015-01-01

    Comet Siding Spring encountered Mars on October 19, 2014 at a distance of about 140,500 km - the nearest comet flyby of a planet in recorded history. Mars Reconnaissance Orbiter (MRO) was able to detect the comet, gather science data, and capture images of the comet as it approached Mars. To help protect MRO from the incoming comet particles, two propulsive maneuvers were performed to position the spacecraft behind Mars at the arrival time of the expected peak particle fluency. This paper documents the strategy that the MRO Navigation Team executed to mitigate risk from the comet particles while allowing scientific observations of the comet flyby.

  18. A new proxy of comet impacts?

    Science.gov (United States)

    Kasatkina, E.; Shumilov, O.; Lukina, N.

    Environmental consequences of impacts by comets and asteroids depend on their sizes. The impact of 10 km sized asteroid killed off the dinosaurs 65 million years ago. A comet impact in AD 536 (>500 m diameter) caused a global climatic downturn. An impact of Tunguska-sized (50 m diameter) object (TO) could devastate a 2000 km -2 forest area, roughly the size of Moscow or New York. Among some misteries of the Tunguska event an important role plays the cause of the accelerated tree growth after 1908 in the catastrophe area. The main explanation is that an accelerated growth of old trees is a result of decrease of the level of competition due to falling of trees. We analyzed tree rings in larch samples collected in the Taymir region of Northern Siberia (72N; 105E) at a distance of about 1500 km to the North from the Tunguska catastrophe epicenter (61N; 102E). It was found that tree ring growth is 150% higher in 1908. We also analyzed all available Siberian tree-ring series (more than 100). It was found that there is a considerable increase of tree ring growth in 1908 over a huge area of Siberia (60N-75N; 60E-110E). It is obvious that the above mentioned interpretation could hardly explain the effect observed. Most likely the tree growth was stimulated by the TO matter spreaded over a large territory of Eurasia and introduced into the soil. It is generally believed that meteorites and comets delivered large amounts of organic to the early Earth. Another hypothesis is connected to NO produced during the Tunguska event. The cometary (probably fragment of the comet Enke) tail started to influence the Earth atmosphere several days before the fall of the meteorite causing optical anomalies observed mainly to the West from the place of TO explosion (Western Siberia, European Russia, Northern Europe). It is interesting that we've discovered nearly the similar, but smaller (about 50%) increase of tree ring growth connected to the Chulym bolide impact in 1984. The Chulym bolide

  19. Workshop on observations of recent comets (1990)

    Energy Technology Data Exchange (ETDEWEB)

    Huebner, W.F.; Wehinger, P.A.; Rahe, J.; Konno, I.

    1990-01-01

    Potential interpretations are presented for observations of four comets: Brorsen-Metcalf (1989o), Okazaki-Levy-Rudenko (1989r), Aarseth-Brewington (1989a1), and Austin (1989o1). The relationship of minor species with each other and possible parents as well as with dust are being pursued in a number of investigations. Of particular interest are the abundance ratios of CH{sub 4} to CO and NH{sub 3} to N{sub 2}. The need for closer collaboration betwen observing teams and modelers is examined. The need for dust size distribution as a function of cometocentric distance to be analyzed in closer collaboration between observers and modelers is discussed.

  20. A lateral electrophoretic flow diagnostic assay.

    Science.gov (United States)

    Lin, Robert; Skandarajah, Arunan; Gerver, Rachel E; Neira, Hector D; Fletcher, Daniel A; Herr, Amy E

    2015-03-21

    Immunochromatographic assays are a cornerstone tool in disease screening. To complement existing lateral flow assays (based on wicking flow) we introduce a lateral flow format that employs directed electrophoretic transport. The format is termed a "lateral e-flow assay" and is designed to support multiplexed detection using immobilized reaction volumes of capture antigen. To fabricate the lateral e-flow device, we employ mask-based UV photopatterning to selectively immobilize unmodified capture antigen along the microchannel in a barcode-like pattern. The channel-filling polyacrylamide hydrogel incorporates a photoactive moiety (benzophenone) to immobilize capture antigen to the hydrogel without a priori antigen modification. We report a heterogeneous sandwich assay using low-power electrophoresis to drive biospecimen through the capture antigen barcode. Fluorescence barcode readout is collected via a low-resource appropriate imaging system (CellScope). We characterize lateral e-flow assay performance and demonstrate a serum assay for antibodies to the hepatitis C virus (HCV). In a pilot study, the lateral e-flow assay positively identifies HCV+ human sera in 60 min. The lateral e-flow assay provides a flexible format for conducting multiplexed immunoassays relevant to confirmatory diagnosis in near-patient settings.

  1. Comet P/Machholtz and the Quadrantid meteor stream

    International Nuclear Information System (INIS)

    Mcintosh, B.A.

    1990-01-01

    Attention is drawn to the suggestive similarities between the calculated perturbation behavior of Comet P/Machholtz 1986 VIII, on the one hand, and on the other those of the Quadrantid, Delta Aquarid, and Arietid meteor streams. There appears to be adequate evidence for the formation by the Comets P/Machholtz and 1491-I, together with the three meteor streams, of a related complex controlled by Jupiter's gravitational perturbations; there is no comparably compelling information, however, bearing on the questions of parent-offspring or sibling relationships among these comets and meteor streams. 13 refs

  2. Development in electrophoresis: instrumentation for two-dimensional gel electrophoresis of protein separation and application of capillary electrophoresis in micro-bioanalysis

    Energy Technology Data Exchange (ETDEWEB)

    Xu, Aoshuang [Iowa State Univ., Ames, IA (United States)

    2008-01-01

    This dissertation begins with a general introduction of topics related to this work. The following chapters contain three scientific manuscripts, each presented in a separate chapter with accompanying tables, figures, and literature citations. The final chapter summarizes the work and provides some prospective on this work. This introduction starts with a brief treatment of the basic principles of electrophoresis separation, followed by a discussion of gel electrophoresis and particularly polyacrylamide gel electrophoresis for protein separation, a summary of common capillary electrophoresis separation modes, and a brief treatment of micro-bioanalysis application of capillary electrophoresis, and ends with an overview of protein conformation and dynamics.

  3. Adapting capillary gel electrophoresis as a sensitive, high-throughput method to accelerate characterization of nucleic acid metabolic enzymes.

    Science.gov (United States)

    Greenough, Lucia; Schermerhorn, Kelly M; Mazzola, Laurie; Bybee, Joanna; Rivizzigno, Danielle; Cantin, Elizabeth; Slatko, Barton E; Gardner, Andrew F

    2016-01-29

    Detailed biochemical characterization of nucleic acid enzymes is fundamental to understanding nucleic acid metabolism, genome replication and repair. We report the development of a rapid, high-throughput fluorescence capillary gel electrophoresis method as an alternative to traditional polyacrylamide gel electrophoresis to characterize nucleic acid metabolic enzymes. The principles of assay design described here can be applied to nearly any enzyme system that acts on a fluorescently labeled oligonucleotide substrate. Herein, we describe several assays using this core capillary gel electrophoresis methodology to accelerate study of nucleic acid enzymes. First, assays were designed to examine DNA polymerase activities including nucleotide incorporation kinetics, strand displacement synthesis and 3'-5' exonuclease activity. Next, DNA repair activities of DNA ligase, flap endonuclease and RNase H2 were monitored. In addition, a multicolor assay that uses four different fluorescently labeled substrates in a single reaction was implemented to characterize GAN nuclease specificity. Finally, a dual-color fluorescence assay to monitor coupled enzyme reactions during Okazaki fragment maturation is described. These assays serve as a template to guide further technical development for enzyme characterization or nucleoside and non-nucleoside inhibitor screening in a high-throughput manner. © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.

  4. Inhibition study of alanine aminotransferase enzyme using sequential online capillary electrophoresis analysis.

    Science.gov (United States)

    Liu, Lina; Chen, Yuanfang; Yang, Li

    2014-12-15

    We report the study of several inhibitors on alanine aminotransferase (ALT) enzyme using sequential online capillary electrophoresis (CE) assay. Using metal ions (Na(+) and Mg(2+)) as example inhibitors, we show that evolution of the ALT inhibition reaction can be achieved by automatically and simultaneously monitoring the substrate consumption and product formation as a function of reaction time. The inhibition mechanism and kinetic constants of ALT inhibition with succinic acid and two traditional Chinese medicines were derived from the sequential online CE assay. Our study could provide valuable information about the inhibition reactions of ALT enzyme. Copyright © 2014 Elsevier Inc. All rights reserved.

  5. Screening for mutations in the uroporphyrinogen decarboxylase gene using denaturing gradient gel electrophoresis

    DEFF Research Database (Denmark)

    Christiansen, L; Ged, C; Hombrados, I

    1999-01-01

    , confirming the heterogeneity of the underlying genetic defects of these diseases. We have established a denaturing gradient gel electrophoresis (DGGE) assay for mutation detection in the UROD gene, enabling the simultaneous screening for known and unknown mutations. The established assay has proved able...... to exon skipping, and a 2-bp deletion (415-416delTA) resulting in a frameshift and the introduction of a premature stop codon. Heterologous expression and enzymatic studies of the mutant proteins demonstrate that the three mutations leading to shortening or truncation of the UROD protein have no residual...

  6. THE PLASMA ENVIRONMENT IN COMETS OVER A WIDE RANGE OF HELIOCENTRIC DISTANCES: APPLICATION TO COMET C/2006 P1 (MCNAUGHT)

    International Nuclear Information System (INIS)

    Shou, Y.; Combi, M.; Gombosi, T.; Toth, G.; Jia, Y.-D.; Rubin, M.

    2015-01-01

    On 2007 January 12, comet C/2006 P1 (McNaught) passed its perihelion at 0.17 AU. Abundant remote observations offer plenty of information on the neutral composition and neutral velocities within 1 million kilometers of the comet nucleus. In early February, the Ulysses spacecraft made an in situ measurement of the ion composition, plasma velocity, and magnetic field when passing through the distant ion tail and the ambient solar wind. The measurement by Ulysses was made when the comet was at around 0.8 AU. With the constraints provided by remote and in situ observations, we simulated the plasma environment of Comet C/2006 P1 (McNaught) using a multi-species comet MHD model over a wide range of heliocentric distances from 0.17 to 1.75 AU. The solar wind interaction of the comet at various locations is characterized and typical subsolar standoff distances of the bow shock and contact surface are presented and compared to analytic solutions. We find the variation in the bow shock standoff distances at different heliocentric distances is smaller than the contact surface. In addition, we modified the multi-species model for the case when the comet was at 0.7 AU and achieved comparable water group ion abundances, proton densities, plasma velocities, and plasma temperatures to the Ulysses/SWICS and SWOOPS observations. We discuss the dominating chemical reactions throughout the comet-solar wind interaction region and demonstrate the link between the ion composition near the comet and in the distant tail as measured by Ulysses

  7. THE PLASMA ENVIRONMENT IN COMETS OVER A WIDE RANGE OF HELIOCENTRIC DISTANCES: APPLICATION TO COMET C/2006 P1 (MCNAUGHT)

    Energy Technology Data Exchange (ETDEWEB)

    Shou, Y.; Combi, M.; Gombosi, T.; Toth, G. [Department of Atmospheric, Oceanic and Space Sciences, University of Michigan, Ann Arbor, MI (United States); Jia, Y.-D. [IGPP, and EPSS, University of California, Los Angeles, CA 90095 (United States); Rubin, M. [Physikalisches Institut, University of Bern, Sidlerstrasse. 5, CH-3012 Bern (Switzerland)

    2015-08-20

    On 2007 January 12, comet C/2006 P1 (McNaught) passed its perihelion at 0.17 AU. Abundant remote observations offer plenty of information on the neutral composition and neutral velocities within 1 million kilometers of the comet nucleus. In early February, the Ulysses spacecraft made an in situ measurement of the ion composition, plasma velocity, and magnetic field when passing through the distant ion tail and the ambient solar wind. The measurement by Ulysses was made when the comet was at around 0.8 AU. With the constraints provided by remote and in situ observations, we simulated the plasma environment of Comet C/2006 P1 (McNaught) using a multi-species comet MHD model over a wide range of heliocentric distances from 0.17 to 1.75 AU. The solar wind interaction of the comet at various locations is characterized and typical subsolar standoff distances of the bow shock and contact surface are presented and compared to analytic solutions. We find the variation in the bow shock standoff distances at different heliocentric distances is smaller than the contact surface. In addition, we modified the multi-species model for the case when the comet was at 0.7 AU and achieved comparable water group ion abundances, proton densities, plasma velocities, and plasma temperatures to the Ulysses/SWICS and SWOOPS observations. We discuss the dominating chemical reactions throughout the comet-solar wind interaction region and demonstrate the link between the ion composition near the comet and in the distant tail as measured by Ulysses.

  8. Contemporary sample stacking in analytical electrophoresis

    Czech Academy of Sciences Publication Activity Database

    Šlampová, Andrea; Malá, Zdeňka; Pantůčková, Pavla; Gebauer, Petr; Boček, Petr

    2013-01-01

    Roč. 34, č. 1 (2013), s. 3-18 ISSN 0173-0835 R&D Projects: GA ČR GAP206/10/1219 Institutional support: RVO:68081715 Keywords : biological samples * stacking * trace analysis * zone electrophoresis Subject RIV: CB - Analytical Chemistry, Separation Impact factor: 3.161, year: 2013

  9. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS ...

    African Journals Online (AJOL)

    Four strains of eri, Samia cynthia ricini Lepidoptera: Saturniidae that can be identified morphologically and maintained at North East Institute of Science and Technology, Jorhat were characterized based on their protein profile by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and DNA by random ...

  10. Preparing size markers for gel electrophoresis.

    Science.gov (United States)

    Nilsen, Timothy W

    2013-12-01

    Here we present two simple methods for preparing radiolabeled size markers for gel electrophoresis. The first procedure describes the generation of an RNA marker ladder by the alkaline hydrolysis of (32)P 5'- or 3'-end-labeled RNA. The second procedure describes the labeling of DNA fragments produced by digestion of pBR322 with the restriction enzyme MspI.

  11. Contemporary sample stacking in analytical electrophoresis

    Czech Academy of Sciences Publication Activity Database

    Malá, Zdeňka; Šlampová, Andrea; Křivánková, Ludmila; Gebauer, Petr; Boček, Petr

    2015-01-01

    Roč. 36, č. 1 (2015), s. 15-35 ISSN 0173-0835 R&D Projects: GA ČR(CZ) GA13-05762S Institutional support: RVO:68081715 Keywords : biological samples * stacking * trace analysis * zone electrophoresis Subject RIV: CB - Analytical Chemistry, Separation Impact factor: 2.482, year: 2015

  12. Electrophoresis test prevalence, requesting patterns, yield and ...

    African Journals Online (AJOL)

    Most of the appropriate SPE test requests were from clinical haematology ... 1 Division of Chemical Pathology, Department of Pathology, National Health Laboratory Service and Faculty of Medicine and Health Sciences,. Stellenbosch ... electrophoresis (IFE)) in a South African (SA) pathology laboratory setting are limited.

  13. DNA DAMAGE QUANTITATION BY ALKALINE GEL ELECTROPHORESIS.

    Energy Technology Data Exchange (ETDEWEB)

    SUTHERLAND,B.M.; BENNETT,P.V.; SUTHERLAND, J.C.

    2004-03-24

    Physical and chemical agents in the environment, those used in clinical applications, or encountered during recreational exposures to sunlight, induce damages in DNA. Understanding the biological impact of these agents requires quantitation of the levels of such damages in laboratory test systems as well as in field or clinical samples. Alkaline gel electrophoresis provides a sensitive (down to {approx} a few lesions/5Mb), rapid method of direct quantitation of a wide variety of DNA damages in nanogram quantities of non-radioactive DNAs from laboratory, field, or clinical specimens, including higher plants and animals. This method stems from velocity sedimentation studies of DNA populations, and from the simple methods of agarose gel electrophoresis. Our laboratories have developed quantitative agarose gel methods, analytical descriptions of DNA migration during electrophoresis on agarose gels (1-6), and electronic imaging for accurate determinations of DNA mass (7-9). Although all these components improve sensitivity and throughput of large numbers of samples (7,8,10), a simple version using only standard molecular biology equipment allows routine analysis of DNA damages at moderate frequencies. We present here a description of the methods, as well as a brief description of the underlying principles, required for a simplified approach to quantitation of DNA damages by alkaline gel electrophoresis.

  14. Fractionation of liver proteins by preparative electrophoresis.

    Science.gov (United States)

    Fountoulakis, M; Juranville, J-F; Tsangaris, G; Suter, L

    2004-02-01

    Proteomics offers unique possibilities to investigate changes in the levels and modifications of proteins involved in the pathomechanisms of diseases and toxic events. However, search for potential drug targets and disease or toxicity markers is limited by the fact that mainly the high-abundance, hydrophilic proteins are visualized in two-dimensional gels. Here we studied the enrichment of rat liver cytosolic proteins by preparative electrophoresis. Preparative electrophoresis was performed with the PrepCell apparatus in the presence of 0.1% lithium dodecyl sulfate. Lithium dodecyl sulfate was exchanged against agents compatible with isoelectric focusing prior to the two-dimensional gel electrophoresis. Proteins were identified from two-dimensional gels by matrix-assisted laser desorption ionization time-of-flight mass specrometry. Low- and middle-size proteins and low-abundance proteins, which had not been found before, were enriched by preparative electrophoresis. The present study represents a contribution of proteomics in the quantification of differences in the levels of low-abundance liver proteins in toxicity studies.

  15. CRUDE PROTEIN ELECTROPHORESIS OF SEEDS OF TEN

    African Journals Online (AJOL)

    A A Essiett

    Seeds of mature fruits of ten species of Solanum were collected from the gardens near the screen house, Botany. Department, Obafemi Awolowo University, Ile Ife, Osun State, Nigeria. Crude seed proteins were extracted from them and characterised using polyacrylamide gel electrophoresis. Inter and intra specific ...

  16. The Meteoroid Fluence at Mars Due to Comet Siding Spring

    Science.gov (United States)

    Moorhead, Althea V.

    2014-01-01

    Long-period comet C/2013 A1 (Siding Spring) is headed for a close encounter with Mars on 2014 Oct 19. A collision between the comet and the planet has been ruled out, but the comets coma may envelop Mars and its man-made satellites. We present an analytic model of the dust component of cometary comae that describes the spatial distribution of cometary dust and meteoroids and their size distribution. If the coma reaches Mars, we estimate a total incident particle fluence on the planet and its satellites of 0.01 particles per square meter. We compare our model with numerical simulations, data from past comet missions, and recent Siding Spring observations.

  17. IHW COMET HALLEY INFRARED SPECTRA, V2.0

    Data.gov (United States)

    National Aeronautics and Space Administration — This data set presents the infrared spectroscopy of Comet 1P/Halley archived with the Infrared Studies Network (IRSN) of the International Halley Watch (IHW). 59...

  18. IHW AMATEUR SPECTROGRAMS OF COMET 1P/HALLEY

    Data.gov (United States)

    National Aeronautics and Space Administration — Report of spectrograms of comet 1P/Halley taken by amateur astronomers and submitted as part of the International Halley Watch Amateur Network. No digital data are...

  19. IHW COMET HALLEY REDUCED SPECTROSCOPIC OBSERVATIONS V1.0

    Data.gov (United States)

    National Aeronautics and Space Administration — NASA's International Halley Watch (IHW) has created a Comet Halley Archive. The collection of data spans the full wavelength range as submitted by scientists to the...

  20. IHW COMET HALLEY INFRARED POLARIMETRY V1.0

    Data.gov (United States)

    National Aeronautics and Space Administration — NASA's International Halley Watch (IHW) has created a Comet Halley Archive. The collection of data spans the full wavelength range as submitted by scientists to the...