Comet Assay: A genotoxicological technique for measuring DNA damage in an individual cell using single-cell gel electrophoresis. Cell DNA fragments assume a "comet with tail" formation on electrophoresis and are detected with an image analysis system. Alkaline assay conditions facilitate sensitive detection of single-strand damage.DNA Damage: Injuries to DNA that introduce deviations from its normal, intact structure and which may, if left unrepaired, result in a MUTATION or a block of DNA REPLICATION. These deviations may be caused by physical or chemical agents and occur by natural or unnatural, introduced circumstances. They include the introduction of illegitimate bases during replication or by deamination or other modification of bases; the loss of a base from the DNA backbone leaving an abasic site; single-strand breaks; double strand breaks; and intrastrand (PYRIMIDINE DIMERS) or interstrand crosslinking. Damage can often be repaired (DNA REPAIR). If the damage is extensive, it can induce APOPTOSIS.DNA Repair: The reconstruction of a continuous two-stranded DNA molecule without mismatch from a molecule which contained damaged regions. The major repair mechanisms are excision repair, in which defective regions in one strand are excised and resynthesized using the complementary base pairing information in the intact strand; photoreactivation repair, in which the lethal and mutagenic effects of ultraviolet light are eliminated; and post-replication repair, in which the primary lesions are not repaired, but the gaps in one daughter duplex are filled in by incorporation of portions of the other (undamaged) daughter duplex. Excision repair and post-replication repair are sometimes referred to as "dark repair" because they do not require light.Micronucleus Tests: Induction and quantitative measurement of chromosomal damage leading to the formation of micronuclei (MICRONUCLEI, CHROMOSOME-DEFECTIVE) in cells which have been exposed to genotoxic agents or IONIZING RADIATION.Mutagens: Chemical agents that increase the rate of genetic mutation by interfering with the function of nucleic acids. A clastogen is a specific mutagen that causes breaks in chromosomes.Mutagenicity Tests: Tests of chemical substances and physical agents for mutagenic potential. They include microbial, insect, mammalian cell, and whole animal tests.DNA-Formamidopyrimidine Glycosylase: A DNA repair enzyme that is an N-glycosyl hydrolase with specificity for DNA-containing ring-opened N(7)-methylguanine residues.DNA Breaks, Single-Stranded: Interruptions in one of the strands of the sugar-phosphate backbone of double-stranded DNA.Gamma Rays: Penetrating, high-energy electromagnetic radiation emitted from atomic nuclei during NUCLEAR DECAY. The range of wavelengths of emitted radiation is between 0.1 - 100 pm which overlaps the shorter, more energetic hard X-RAYS wavelengths. The distinction between gamma rays and X-rays is based on their radiation source.DNA Breaks, Double-Stranded: Interruptions in the sugar-phosphate backbone of DNA, across both strands adjacently.Ataxia Telangiectasia Mutated Proteins: A group of PROTEIN-SERINE-THREONINE KINASES which activate critical signaling cascades in double strand breaks, APOPTOSIS, and GENOTOXIC STRESS such as ionizing ultraviolet A light, thereby acting as a DNA damage sensor. These proteins play a role in a wide range of signaling mechanisms in cell cycle control.Deoxyguanosine: A nucleoside consisting of the base guanine and the sugar deoxyribose.Antimutagenic Agents: Agents that reduce the frequency or rate of spontaneous or induced mutations independently of the mechanism involved.Oxidative Stress: A disturbance in the prooxidant-antioxidant balance in favor of the former, leading to potential damage. Indicators of oxidative stress include damaged DNA bases, protein oxidation products, and lipid peroxidation products (Sies, Oxidative Stress, 1991, pxv-xvi).DNA Breaks: Interruptions in the sugar-phosphate backbone of DNA.DNA: A deoxyribonucleotide polymer that is the primary genetic material of all cells. Eukaryotic and prokaryotic organisms normally contain DNA in a double-stranded state, yet several important biological processes transiently involve single-stranded regions. DNA, which consists of a polysugar-phosphate backbone possessing projections of purines (adenine and guanine) and pyrimidines (thymine and cytosine), forms a double helix that is held together by hydrogen bonds between these purines and pyrimidines (adenine to thymine and guanine to cytosine).Methyl Methanesulfonate: An alkylating agent in cancer therapy that may also act as a mutagen by interfering with and causing damage to DNA.Ultraviolet Rays: That portion of the electromagnetic spectrum immediately below the visible range and extending into the x-ray frequencies. The longer wavelengths (near-UV or biotic or vital rays) are necessary for the endogenous synthesis of vitamin D and are also called antirachitic rays; the shorter, ionizing wavelengths (far-UV or abiotic or extravital rays) are viricidal, bactericidal, mutagenic, and carcinogenic and are used as disinfectants.Meteoroids: Any solid objects moving in interplanetary space that are smaller than a planet or asteroid but larger than a molecule. Meteorites are any meteoroid that has fallen to a planetary surface. (From McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)Lymphocytes: White blood cells formed in the body's lymphoid tissue. The nucleus is round or ovoid with coarse, irregularly clumped chromatin while the cytoplasm is typically pale blue with azurophilic (if any) granules. Most lymphocytes can be classified as either T or B (with subpopulations of each), or NATURAL KILLER CELLS.Cell Cycle Proteins: Proteins that control the CELL DIVISION CYCLE. This family of proteins includes a wide variety of classes, including CYCLIN-DEPENDENT KINASES, mitogen-activated kinases, CYCLINS, and PHOSPHOPROTEIN PHOSPHATASES as well as their putative substrates such as chromatin-associated proteins, CYTOSKELETAL PROTEINS, and TRANSCRIPTION FACTORS.Dose-Response Relationship, Radiation: The relationship between the dose of administered radiation and the response of the organism or tissue to the radiation.Micronuclei, Chromosome-Defective: Defective nuclei produced during the TELOPHASE of MITOSIS or MEIOSIS by lagging CHROMOSOMES or chromosome fragments derived from spontaneous or experimentally induced chromosomal structural changes.Cosmic Dust: Finely divided solid matter with particle sizes smaller than a micrometeorite, thus with diameters much smaller than a millimeter, moving in interplanetary space. (NASA Thesaurus, 1994)Radiation Tolerance: The ability of some cells or tissues to survive lethal doses of IONIZING RADIATION. Tolerance depends on the species, cell type, and physical and chemical variables, including RADIATION-PROTECTIVE AGENTS and RADIATION-SENSITIZING AGENTS.Hydrogen Peroxide: A strong oxidizing agent used in aqueous solution as a ripening agent, bleach, and topical anti-infective. It is relatively unstable and solutions deteriorate over time unless stabilized by the addition of acetanilide or similar organic materials.Tumor Suppressor Protein p53: Nuclear phosphoprotein encoded by the p53 gene (GENES, P53) whose normal function is to control CELL PROLIFERATION and APOPTOSIS. A mutant or absent p53 protein has been found in LEUKEMIA; OSTEOSARCOMA; LUNG CANCER; and COLORECTAL CANCER.DNA Fragmentation: Splitting the DNA into shorter pieces by endonucleolytic DNA CLEAVAGE at multiple sites. It includes the internucleosomal DNA fragmentation, which along with chromatin condensation, are considered to be the hallmarks of APOPTOSIS.DNA-Binding Proteins: Proteins which bind to DNA. The family includes proteins which bind to both double- and single-stranded DNA and also includes specific DNA binding proteins in serum which can be used as markers for malignant diseases.Apoptosis: One of the mechanisms by which CELL DEATH occurs (compare with NECROSIS and AUTOPHAGOCYTOSIS). Apoptosis is the mechanism responsible for the physiological deletion of cells and appears to be intrinsically programmed. It is characterized by distinctive morphologic changes in the nucleus and cytoplasm, chromatin cleavage at regularly spaced sites, and the endonucleolytic cleavage of genomic DNA; (DNA FRAGMENTATION); at internucleosomal sites. This mode of cell death serves as a balance to mitosis in regulating the size of animal tissues and in mediating pathologic processes associated with tumor growth.Cell Survival: The span of viability of a cell characterized by the capacity to perform certain functions such as metabolism, growth, reproduction, some form of responsiveness, and adaptability.Cell Cycle: The complex series of phenomena, occurring between the end of one CELL DIVISION and the end of the next, by which cellular material is duplicated and then divided between two daughter cells. The cell cycle includes INTERPHASE, which includes G0 PHASE; G1 PHASE; S PHASE; and G2 PHASE, and CELL DIVISION PHASE.Neon: Neon. A noble gas with the atomic symbol Ne, atomic number 10, and atomic weight 20.18. It is found in the earth's crust and atmosphere as an inert, odorless gas and is used in vacuum tubes and incandescent lamps.Histones: Small chromosomal proteins (approx 12-20 kD) possessing an open, unfolded structure and attached to the DNA in cell nuclei by ionic linkages. Classification into the various types (designated histone I, histone II, etc.) is based on the relative amounts of arginine and lysine in each.Protein-Serine-Threonine Kinases: A group of enzymes that catalyzes the phosphorylation of serine or threonine residues in proteins, with ATP or other nucleotides as phosphate donors.Checkpoint Kinase 2: Enzyme activated in response to DNA DAMAGE involved in cell cycle arrest. The gene is located on the long (q) arm of chromosome 22 at position 12.1. In humans it is encoded by the CHEK2 gene.Spermatozoa: Mature male germ cells derived from SPERMATIDS. As spermatids move toward the lumen of the SEMINIFEROUS TUBULES, they undergo extensive structural changes including the loss of cytoplasm, condensation of CHROMATIN into the SPERM HEAD, formation of the ACROSOME cap, the SPERM MIDPIECE and the SPERM TAIL that provides motility.Ethyl Methanesulfonate: An antineoplastic agent with alkylating properties. It also acts as a mutagen by damaging DNA and is used experimentally for that effect.Cell Line, Tumor: A cell line derived from cultured tumor cells.Tumor Suppressor Proteins: Proteins that are normally involved in holding cellular growth in check. Deficiencies or abnormalities in these proteins may lead to unregulated cell growth and tumor development.Leukemia L5178: An experimental lymphocytic leukemia of mice.DNA Adducts: The products of chemical reactions that result in the addition of extraneous chemical groups to DNA.Reactive Oxygen Species: Molecules or ions formed by the incomplete one-electron reduction of oxygen. These reactive oxygen intermediates include SINGLET OXYGEN; SUPEROXIDES; PEROXIDES; HYDROXYL RADICAL; and HYPOCHLOROUS ACID. They contribute to the microbicidal activity of PHAGOCYTES, regulation of signal transduction and gene expression, and the oxidative damage to NUCLEIC ACIDS; PROTEINS; and LIPIDS.Antioxidants: Naturally occurring or synthetic substances that inhibit or retard the oxidation of a substance to which it is added. They counteract the harmful and damaging effects of oxidation in animal tissues.DNA Glycosylases: A family of DNA repair enzymes that recognize damaged nucleotide bases and remove them by hydrolyzing the N-glycosidic bond that attaches them to the sugar backbone of the DNA molecule. The process called BASE EXCISION REPAIR can be completed by a DNA-(APURINIC OR APYRIMIDINIC SITE) LYASE which excises the remaining RIBOSE sugar from the DNA.Electrophoresis, Agar Gel: Electrophoresis in which agar or agarose gel is used as the diffusion medium.Radiation: Emission or propagation of acoustic waves (SOUND), ELECTROMAGNETIC ENERGY waves (such as LIGHT; RADIO WAVES; GAMMA RAYS; or X-RAYS), or a stream of subatomic particles (such as ELECTRONS; NEUTRONS; PROTONS; or ALPHA PARTICLES).X-Rays: Penetrating electromagnetic radiation emitted when the inner orbital electrons of an atom are excited and release radiant energy. X-ray wavelengths range from 1 pm to 10 nm. Hard X-rays are the higher energy, shorter wavelength X-rays. Soft x-rays or Grenz rays are less energetic and longer in wavelength. The short wavelength end of the X-ray spectrum overlaps the GAMMA RAYS wavelength range. The distinction between gamma rays and X-rays is based on their radiation source.7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide: 7,8,8a,9a-Tetrahydrobenzo(10,11)chryseno (3,4-b)oxirene-7,8-diol. A benzopyrene derivative with carcinogenic and mutagenic activity.Cell Line: Established cell cultures that have the potential to propagate indefinitely.Dose-Response Relationship, Drug: The relationship between the dose of an administered drug and the response of the organism to the drug.Cells, Cultured: Cells propagated in vitro in special media conducive to their growth. Cultured cells are used to study developmental, morphologic, metabolic, physiologic, and genetic processes, among others.4-Nitroquinoline-1-oxide: A potent mutagen and carcinogen. This compound and its metabolite 4-HYDROXYAMINOQUINOLINE-1-OXIDE bind to nucleic acids. It inactivates bacteria but not bacteriophage.Casearia: A plant genus of the family FLACOURTIACEAE. Members contain casearins which are clerodane type DITERPENES.DNA Repair Enzymes: Enzymes that are involved in the reconstruction of a continuous two-stranded DNA molecule without mismatch from a molecule, which contained damaged regions.Nuclear Proteins: Proteins found in the nucleus of a cell. Do not confuse with NUCLEOPROTEINS which are proteins conjugated with nucleic acids, that are not necessarily present in the nucleus.Occupational Exposure: The exposure to potentially harmful chemical, physical, or biological agents that occurs as a result of one's occupation.Radiation, Ionizing: ELECTROMAGNETIC RADIATION or particle radiation (high energy ELEMENTARY PARTICLES) capable of directly or indirectly producing IONS in its passage through matter. The wavelengths of ionizing electromagnetic radiation are equal to or smaller than those of short (far) ultraviolet radiation and include gamma and X-rays.HeLa Cells: The first continuously cultured human malignant CELL LINE, derived from the cervical carcinoma of Henrietta Lacks. These cells are used for VIRUS CULTIVATION and antitumor drug screening assays.Mutation: Any detectable and heritable change in the genetic material that causes a change in the GENOTYPE and which is transmitted to daughter cells and to succeeding generations.Phosphorylation: The introduction of a phosphoryl group into a compound through the formation of an ester bond between the compound and a phosphorus moiety.Poly(ADP-ribose) Polymerases: Enzymes that catalyze the transfer of multiple ADP-RIBOSE groups from nicotinamide-adenine dinucleotide (NAD) onto protein targets, thus building up a linear or branched homopolymer of repeating ADP-ribose units i.e., POLY ADENOSINE DIPHOSPHATE RIBOSE.G2 Phase: The period of the CELL CYCLE following DNA synthesis (S PHASE) and preceding M PHASE (cell division phase). The CHROMOSOMES are tetraploid in this point.Alkylating Agents: Highly reactive chemicals that introduce alkyl radicals into biologically active molecules and thereby prevent their proper functioning. Many are used as antineoplastic agents, but most are very toxic, with carcinogenic, mutagenic, teratogenic, and immunosuppressant actions. They have also been used as components in poison gases.Ethylnitrosourea: A nitrosourea compound with alkylating, carcinogenic, and mutagenic properties.Radiation-Protective Agents: Drugs used to protect against ionizing radiation. They are usually of interest for use in radiation therapy but have been considered for other, e.g. military, purposes.Carcinogens: Substances that increase the risk of NEOPLASMS in humans or animals. Both genotoxic chemicals, which affect DNA directly, and nongenotoxic chemicals, which induce neoplasms by other mechanism, are included.Chromosome Aberrations: Abnormal number or structure of chromosomes. Chromosome aberrations may result in CHROMOSOME DISORDERS.Genomic Instability: An increased tendency of the GENOME to acquire MUTATIONS when various processes involved in maintaining and replicating the genome are dysfunctional.Agrochemicals: Chemicals used in agriculture. These include pesticides, fumigants, fertilizers, plant hormones, steroids, antibiotics, mycotoxins, etc.Plant Extracts: Concentrated pharmaceutical preparations of plants obtained by removing active constituents with a suitable solvent, which is evaporated away, and adjusting the residue to a prescribed standard.Melastomataceae: A plant family of the order Myrtales, subclass Rosidae, class Magnoliopsida composed of tropical plants with parallel-nerved leaves.Fibroblasts: Connective tissue cells which secrete an extracellular matrix rich in collagen and other macromolecules.Time Factors: Elements of limited time intervals, contributing to particular results or situations.Mitomycin: An antineoplastic antibiotic produced by Streptomyces caespitosus. It is one of the bi- or tri-functional ALKYLATING AGENTS causing cross-linking of DNA and inhibition of DNA synthesis.Benzo(a)pyrene: A potent mutagen and carcinogen. It is a public health concern because of its possible effects on industrial workers, as an environmental pollutant, an as a component of tobacco smoke.Coke: A residue of coal, left after dry (destructive) distillation, used as a fuel.DNA Replication: The process by which a DNA molecule is duplicated.Cell Nucleus: Within a eukaryotic cell, a membrane-limited body which contains chromosomes and one or more nucleoli (CELL NUCLEOLUS). The nuclear membrane consists of a double unit-type membrane which is perforated by a number of pores; the outermost membrane is continuous with the ENDOPLASMIC RETICULUM. A cell may contain more than one nucleus. (From Singleton & Sainsbury, Dictionary of Microbiology and Molecular Biology, 2d ed)Cell Death: The termination of the cell's ability to carry out vital functions such as metabolism, growth, reproduction, responsiveness, and adaptability.DNA-Activated Protein Kinase: A serine-threonine protein kinase that, when activated by DNA, phosphorylates several DNA-binding protein substrates including the TUMOR SUPPRESSOR PROTEIN P53 and a variety of TRANSCRIPTION FACTORS.GuanineLeukocytes: White blood cells. These include granular leukocytes (BASOPHILS; EOSINOPHILS; and NEUTROPHILS) as well as non-granular leukocytes (LYMPHOCYTES and MONOCYTES).Bromates: Negative ions or salts derived from bromic acid, HBrO3.Struthioniformes: An order of flightless birds comprising the ostriches, which naturally inhabit open, low rainfall areas of Africa.Oxidation-Reduction: A chemical reaction in which an electron is transferred from one molecule to another. The electron-donating molecule is the reducing agent or reductant; the electron-accepting molecule is the oxidizing agent or oxidant. Reducing and oxidizing agents function as conjugate reductant-oxidant pairs or redox pairs (Lehninger, Principles of Biochemistry, 1982, p471).Medical Secretaries: Individuals responsible for various duties pertaining to the medical office routine.Polycyclic Hydrocarbons, Aromatic: A major group of unsaturated cyclic hydrocarbons containing two or more rings. The vast number of compounds of this important group, derived chiefly from petroleum and coal tar, are rather highly reactive and chemically versatile. The name is due to the strong and not unpleasant odor characteristic of most substances of this nature. (From Hawley's Condensed Chemical Dictionary, 12th ed, p96)DNA, Neoplasm: DNA present in neoplastic tissue.Lipid Peroxidation: Peroxidase catalyzed oxidation of lipids using hydrogen peroxide as an electron acceptor.
... by comet assay, advanced maternal age and semen quality. A systematic meta-analysis of 24 estimates of DNA damage based on a ... PGD is a process in which one or two cells from an embryo on Day 3 or Day 5 are extracted and the cells genetically analyzed. ... "A systematic review and meta-analysis to determine the effect of sperm DNA damage on in vitro fertilization and ... Basic research on the maturation of sperm shows that hyaluronan-binding sperm are more mature and show fewer DNA strand breaks ...
... and Comet Assay have been developed to assess the chemicals' potential to cause DNA damage that may lead to cancer. The ... To further analyze the site of lesion, it was observed that polymerase stopped at the site and adenine was inappropriately ... To assay for genotoxic molecules, researchers assay for DNA damage in cells exposed to the toxic substrates. This DNA damage ... By utilizing the destructive properties of genotoxins treatments aims to induce DNA damage into cancer cells. Any damage done ...
... and Comet Assay have been developed to assess the chemicals' potential to cause DNA damage that may lead to cancer. ... To further analyze the site of lesion, it was observed that polymerase stopped at the site and adenine was inappropriately ... To assay for genotoxic molecules, researchers assay for DNA damage in cells exposed to the toxic substrates. This DNA damage ... Comet assayEdit. See also: Comet assay. Comet assays are one of the most common tests for genotoxicity. The technique involves ...
The damage to the DNA was measured for individual cells using single-gel electrophoresis via the comet assay. Researchers have ... A complete study analyzing the effect of varying sized platinum nanoparticles used both in vivo and in vitro models is used to ... The 15 mg/kg dose of sub 1 nm platinum nanoparticles were found to cause liver damage while the larger particles had no effect ... A nanoparticle can also interact with the cell's DNA or genome to cause genotoxicity. These effects are seen in different ...
... the Comet assay or by the TUNEL assay. Men with sperm motility defects often have high levels of sperm DNA fragmentation. The ... and TUNEL assay are both effective in detecting sperm DNA damage. Using bright-field microscopy, the SCD test appears to be ... is a technique for analyzing the variable lengths of DNA fragments that result from digesting a DNA sample with a restriction ... The DNA pass through a gauge needle several times to physically tear the DNA into fine pieces. French pressure cells pass DNA ...
Swain, U; Subba Rao, K (Aug 2011). "Study of DNA damage via the comet assay and base excision repair activities in rat brain ... Genomic data from about 70,000 women were analyzed to identify protein-coding variation associated with age at natural ... DNA damages and mutations are related because DNA damages often cause errors of DNA synthesis during replication or repair and ... If a DNA repair protein is deficient, unrepaired DNA damages tend to accumulate. Such accumulated DNA damages appear to cause ...
... by comet assay, advanced maternal age and semen quality. A systematic meta-analysis of 24 estimates of DNA damage based on a ... PGD is a process in which one or two cells from an embryo on Day 3 or Day 5 are extracted and the cells genetically analyzed. ... "A systematic review and meta-analysis to determine the effect of sperm DNA damage on in vitro fertilization and ... However, potential factors that may influence pregnancy rates (and live birth rates) in ICSI include level of DNA fragmentation ...
For example, the percentage of sperm with highly damaged DNA, comet extent, DNA break number, and other comet measures has been ... Enzyme-linked immunosorbent assay, Radiolabelled Antiglobulin Assay. One of the most informative and specific tests is ... A semen analysis typically measures the number of sperm per millilitre of ejaculate, and analyzes the morphology (shape) and ... Daily sexual activity increases sperm quality in men minimizing DNA damage in the sperm -because it is speculated to result in ...
Effect of oridonin on radiation-induced DNA damage. (A) Representative images and quantification of DNA comet assays. Comet ... protein levels were analyzed by Western blot. Ten µg of protein was loaded per lane (upper panel). Band intensity was ... DNA damage, and apoptosis assays. In vitro, oridonin enhanced the radiation-induced inhibition of cell growth and clonogenic ... Oridonin Enhances Radiation-Induced Cell Death by Promoting DNA Damage in Non-Small Cell Lung Cancer Cells.. Park H1, Jeong YJ2 ...
... mRNA transport and DNA damage response. In this study, we analyzed pathophysiological alterations associated with ALS related ... The accumulation of DNA damage and the cellular response to DNA damage stressors was more pronounced in post-mitotic mFUS ... mRNA transport and DNA damage response. In this study, we analyzed pathophysiological alterations associated with ALS related ... Moreover, even in non-stressed post-mitotic mFUS motoneurons clear signs of DNA-damage could be detected. In summary, we found ...
8-OHdG production of genomic DNA was determined by ELISA (E). DNA strand breaks were analyzed by comet assays (F); DNA damage ... DNA strand breaks (N) in individual cells were analyzed by comet assays. Genomic DNA was used for analysis of oxidative DNA ... DNA damage extent was analyzed by comet assays, and quantified by the mean tail moment (I).. (J and K) PGC-1α+/+ApoE−/− mice (5 ... comet assays). DNA damage extent was shown as the visual scoring units and tail moment.. (K-P) PGC-1α−/−ApoE−/− MASMs were ...
DNA damage was induced by exposing the cells to 4mM H2O2 for 15 minutes. This damage was then measured by the comet assay. We ... in order to determine the DNA repair rate. The results were analyzed with Tri-Tek Comet Score Software Version 1.5. The DNA ... We measured the DNA repair rate of cells by the Comet Assay, taken at several time points throughout the course of 1 hour. ... is itself fuelled by DNA damage and often by errors made by the DNA repair machinery. DNA repair, using natural sources of ...
Effect of cigarette smoking on DNA damage of human cumulus cells analyzed by comet assay. Reprod Toxicol. 2005;20:65-71. [ ... Comet assay. DNA single- and double-strand breaks were detected by alkaline and neutral comet assays, respectively, as ... and double-strand DNA damage and subsequent repair.. The principle of the comet assay is based on alterations found in DNA such ... Cigarette smoke-induced DNA damage and repair detected by the comet assay in HPV-transformed cervical cells. Afsoon Moktar,1 ...
... and DNA damage was analyzed by the alkaline comet assay. Significantly higher levels of DNA damage (strand breaks) were found ... 13] Azqueta A., Collins A.R. The essential comet assay: a comprehensive guide to measuring DNA damage and repair, Arch Toxicol ... and comet assay analysis of DNA damage in whole blood, Free Radic Biol Med., 2011, 51, 719-725. Web of ScienceGoogle Scholar ... isolated from simply frozen whole blood can be used in human biomonitoring for DNA damage measurement with the comet assay, ...
To analyze reproductive hormone levels and neutral comet assays for sperm DNA damage, the authors used enzyme immunoassays. ...
The comet assay is an efficient method to detect DNA damage including single and double-stranded DNA breaks. We describe ... "comet tail"). With nucleotide staining, the extent of DNA damage can be quantified by analyzing "comets" formed by this single ... In Vivo Alkaline Comet Assay and Enzyme-modified Alkaline Comet Assay for Measuring DNA Strand Breaks and Oxidative DNA Damage ... In Vivo Alkaline Comet Assay and Enzyme-modified Alkaline Comet Assay for Measuring DNA Strand Breaks and Oxidative DNA Damage ...
Comet assays revealed higher DNA damage in both epididymal and testicular sperm from these mice relative to controls, with ... 100 DNA tails were photographed and analyzed per slide. The length of each tail was measured from the center of the comet to ... Comet assay. Comet assay was performed using the Trevigen Kit (Trevigen, Gaithersburg, MD, USA, cat. no. 4250-050-K) under ... and each tail was categorized into one of four tail types reflecting the severity of DNA damage. The severity of DNA damage ...
... is a sensitive and rapid technique for quantifying and analyzing DNA damage in individual cells. ... The Comet Assay, also called single cell gel electrophoresis (SCGE), ... oxidative base damage, and DNA cross-linking with DNA or protein. The Comet Assay is also used to monitor DNA repair by living ... The Comet Assay can be used to detect DNA damage caused by double strand breaks, single strand breaks, alkali labile sites, ...
Vitamin C significantly decreased ROS, DNA damage, TNF-α, and IL-6. Vitamin C inhibited LPS-induced ROS, DNA damage, TNF-α, IL- ... and IL-6 were analyzed in CAP patients and LPS-stimulated macrophages cells. MH-S cells were transfected with RFP-LC3 plasmids ... Severe CAP patients showed significantly increased ROS, DNA damage, TNF-α, and IL-6. SOD was significantly decreased in severe ... These findings indicated that severe CAP exhibited significantly increased oxidative stress, DNA damage, and proinflammatory ...
Furthermore, with the comet assay, a reduction in DNA damage was observed and statistically confirmed in the greater number of ... The data for the micronucleus and comet assays obtained in three replicate tests were analyzed statistically by ANOVA and by ... The comet assay (Single Cell Gel Test) - A sensitive genotoxicity test for the detection of DNA damage and repair. In: ... Comet test. The results obtained in relation to genotoxicity assessment of the organic extracts of A. blazei in comet assay ( ...
In the comet assay single cells are analyzed with respect to their level of DNA damage. Discrimination of the individual cell ... Reference cells and ploidy in the comet assay Brunborg, Gunnar; Collins, Andrew Richard; Graupner, Anne; Gutzkow, Kristine ... or cell type based on DNA content, with concomitant scoring of the DNA damage, is useful since ... ... Astrometry, Morphology, and Polarimetry of Comet Donati in 1858 Pettersen, Bjørn Ragnvald (Journal article / Tidsskriftartikkel ...
In the comet assay single cells are analyzed with respect to their level of DNA damage. Discrimination of the individual cell ... Reference cells and ploidy in the comet assay  Brunborg, Gunnar; Collins, Andrew Richard; Graupner, Anne; Gutzkow, Kristine ... or cell type based on DNA content, with concomitant scoring of the DNA damage, is useful since ... ...
DNA damage in lens epithelial cells and in lymphocytes analyzed by the comet assay The DNA damage caused by γ-ray (between 0 ... DNA damage and repair were evaluated by alkaline comet assay and γH2AX foci assay. Using the comet assay, we observed a dose- ... Comet assay: DNA damage in lens epithelial cells and lymphocytes after irradiation. a The damage of DNA (as indicated by the ... Examples of lens epithelial cells after irradiation and comet assay. Examples of DNA damage after comet technique in mouse lens ...
Olive P. (1999). DNA damage and repair in individual cells: applications of the comet assay in radiobiology. Int. J. Radiat. ... Table I. DNA damage CHO cells analyzed immediately after exposure to 50 mSv x-rays. Standard error of the mean is indicated ... 9. Collins A. (2004). The comet assay for DNA damage repair. Mol. Biotechnol. 26:249-260. [ Links ]. ... 1). In order to quantify DNA damage from the comets, the measuring method of Collins (2004) was used. Index Damage (ID) was ...
To evaluate the biological mechanisms underlying skin damage caused by fine particulate matter (PM2.5... ... i DNA damage was evaluated by the Comet assay; representative images show comet tails and the graph shows quantification of ... cellular DNA damage. j Lipid peroxidation was analyzed by confocal microscopy after DPPP staining. k Protein oxidation was ... 1h). The Comet assay assessing DNA breakage indicated that PM2.5 increased the presence of cellular DNA tails by 30% compared ...
Single Cell Gel Electrophoresis Assay (Comet Assay) was applied to peripheral blood lymphocytes of five groups of children ... PAH samples were analyzed following US/EPA TO-13-A method. Results indicated the presence in the air of most of the 16 PAHs ... The purpose of this study was to quantify the levels of PAHs in the air at several points in the state, and to analyze their ... relationship with possible damage to DNA on local inhabitants. ... Single Cell Gel Electrophoresis Assay (Comet Assay) was applied ...
Fifty cells per slide were analyzed and scored in triplicate using Comet Assay Software Project (CASP) image analysis software ... Comet Assay for DNA Damage. Single cell gel electrophoresis (SCGE), also known as the alkaline comet assay, was used to measure ... To assess DNA damage products, SH-SY5Y cells were subjected to comet assays. This method measures the ability of damaged DNA to ... G. Wang and K. M. Vasquez, "Impact of alternative DNA structures on DNA damage, DNA repair, and genetic instability," DNA ...
The CometAssay Analysis Software has automated comet acquisition and scoring that can analyze more than half a million comets ... Variability between the CometChip assays are less than 10%, with no overlapping comets requiring manual editing. The well-to- ... DNA Damage and Repair Kits & Reagents. CometChip Brochure. CometAssay System Brochure. CometAssay Analysis Software Brochure. ... Advances in High-Throughput DNA Damage Detection Using CometAssay® and CometChip® Technologies. Register to View Webinar ...
Clonogenic survival assays and comet assays also show that Spy1 expression enhances cell survival after DNA damage. Consistent ... Lysates were analyzed by 10% SDS-PAGE followed by immunoblotting with Myc antisera to detect Myc-Spy1 expression. C, DNA damage ... comet assays demonstrated that Spy1-expressing cells exhibit significantly less damaged DNA after CPT-induced DNA damage. These ... B, quantitation of damaged DNA in mock and Spy1-expressing cells exposed to CPT as measured by the alkaline comet assay. ...
The effects of noise on DNA integrity in all three brain areas were evaluated by using Comet assay. In parallel studies, brain ... hippocampus and cerebellum were analyzed at different time intervals (24 h and 7 days) after noise exposure. Loud noise ... The effects of noise on DNA integrity in all three brain areas were evaluated by using Comet assay. In parallel studies, brain ... Loud noise produced a sudden increase in DNA damage in all the brain areas under investigation. Monoamine levels detected at 7 ...
Products & Services / Cell Stress and DNA Damage / DNA Damage / Comet Assay / Comet Analysis Software / Comet Analysis Software ... It is compatible with 4X images which allows for rapid screening of up to 300 comets in seconds when analyzing CometChip®. ... to automatically locate and score comets to characterize and quantify the degree of DNA damage revealed by the comet assay. ... Alkaline or Neutral Comet. *Measurements for Percent DNA in Tail, Tail Moment, Tail Length, Integrated Cell Intensity, Tail ...
Effect of cigarette smoking on DNA damage of human cumulus cells analyzed by comet assay. Reprod Toxicol 2005;20(1):65-71. ... Polycyclic aromatic hydrocarbon-DNA adducts in human sperm as a marker of DNA damage and infertility. Mutat Res 2003;535(2):155 ... Jeng HA, Pan C-H, Chao M-R, Lin W-Y. Sperm DNA oxidative damage and DNA adducts. Mutat Res 2015;794:75-82. ... Bulky DNA adducts in human sperm associated with semen parameters and sperm DNA fragmentation in infertile men: a cross- ...
Comet Assay - Free download as Powerpoint Presentation (.ppt), PDF File (.pdf), Text File (.txt) or view presentation slides ... is a sensitive and rapid technique for quantifying and analyzing DNA damage in individual cells. This is one of the techniques ... The extent of DNA liberated from the head of the comet is directly proportional to the amount of DNA damage. The Comet Assay ... oxidative base damage, and DNA cross-linking with DNA or protein. The Comet Assay is also used to monitor DNA repair by living ...
  • In both age groups, relationships between endogenous exposure to organochloride substances (polychlorinated biphenyl, hexachlorobenzene, dichlorodiphenyl dichloroethane), metals (cadmium, lead), and urinary metabolites (1-hydroxypyrene, trans-trans muconic acid) versus genotoxic effects (Comet assay and micronuclei in lymphocytes, and urinary 8-hydroxydeoxyguanosine) were investigated. (aacrjournals.org)
  • Several in vivo mutagenicity studies with 1,2-dichloroethane are available in which endpoints such as induction of micronuclei (MN), sister chromatid exchanges (SCE), germ cell mutations, DNA-breakage and sex-linked-recessive-lethal mutations, somatic mutations and recombinations in Drosophila melanogaster were investigated. (europa.eu)
  • Comet assay and BMCyt (micronuclei and nuclear buds) data revealed DNA damage in soybean workers. (thefoodpirate.org)
  • The researchers concluded from testing that when metabolically activated, "PAs produce DNA adducts, DNA cross-linking, DNA breaks, sister chromatid exchange, micronuclei, chromosomal aberrations, gene mutations, and chromosome mutations in vivo and in vitro. (wikipedia.org)
  • The FUS gene codes for a multifunctional RNA/DNA-binding protein that is primarily localized in the nucleus and is involved in cellular processes such as splicing, translation, mRNA transport and DNA damage response. (frontiersin.org)
  • This decisive signaling network includes ATM (ataxia telangiectasia mutated) and ATR (ataxia telangiectasia and Rad3 related), which transduce the DNA damage signal, and the effector UV-DDB (UV-damaged DNA binding protein complex), which elicits a DNA repair response. (asm.org)
  • In order to assess DNA damage associated with exposure to environmental pollution in two polluted sites and one control site in Kosovo, whole blood samples were collected from volunteers in two polluted areas (Kastriot/ Obiliq - lignite-based power plants and lignite mines - and Drenas/Gllogovc - Ferronikeli smelting plant) as well as from Peja, representing an unpolluted area. (degruyter.com)
  • Comet assay responses as indicators of carcinogen exposure. (sigmaaldrich.com)
  • 2017 ) and mitochondrial damage may be a critical part of the pathophysiological mechanisms induced by PM 2.5 exposure. (springer.com)
  • In this study we investigated DNA, neurotransmitters and immune-histochemical alterations induced by exposure to loud noise in three major brain areas (cerebellum, hippocampus, striatum) of Wistar rats. (frontiersin.org)
  • In parallel studies, brain monoamine levels and morphology of nigrostriatal pathways, hippocampus and cerebellum were analyzed at different time intervals (24 h and 7 days) after noise exposure. (frontiersin.org)
  • Loud noise exposure lasting 12 h causes immediate DNA, and long-lasting neurotransmitter and immune-histochemical alterations within specific brain areas of the rat. (frontiersin.org)
  • This study aims to investigate whether low-level hexavalent chromium (Cr(VI)) exposure can cause DNA damage in electroplating workers. (biomedcentral.com)
  • Low-level occupational chromium exposure induced DNA damage. (biomedcentral.com)
  • Oxidative stress and DNA damage have been proposed as mechanisms linking pesticide exposure to health effects such as cancer and neurological diseases. (cdc.gov)
  • A pilot study of pesticide applicators and farm workers working in the fruit orchards of Oregon (i.e., apples, pears) was conducted to examine the relationship between organophosphate (OP) pesticide exposure and oxidative stress and DNA damage. (cdc.gov)
  • Titanium dioxide nanoparticles exposure causes toxic effects on the lung and administration of Idebenone offers protection against its damaging effects. (academicjournals.org)
  • The comet assay results indicated that upon exposure to the same dose of chemical mutagen, CRY2 - cells accumulate significantly more unrepaired DNA damage than CRY2 + cells (P = 0.040), suggesting that CRY2 may be important for DNA repair. (biomedcentral.com)
  • Click to enlarge) Subham conducts an ethoxyresorufin-O-deethylase or EROD assay to measure the activity of the detoxifying enzyme CYP1A1 under PAH exposure. (gulfresearchinitiative.org)
  • His work in her lab initially focused on analyzing the impacts of oxygenated polycyclic aromatic hydrocarbon (PAH) exposure on the embryos of Japanese rice fish (also known as the medaka or Japanese killifish Oryzias latipes ). (gulfresearchinitiative.org)
  • The extent of damage, measured as tail moment, was proportional to exposure duration and was reduced by preincubation with sodium azide. (arvojournals.org)
  • EAC risks were further increased among individuals with both net mutagen-induced DNA damages and exposure to gastroesophageal reflux disease or smoking, known risk factors for EAC. (omicsonline.org)
  • Further, the ubiquitous nature of BPA allows continuous exposure of the human genome concurrent with the normal endogenous and exogenous insults to the genome, and this co-exposure may impact the DNA damage response and repair. (jove.com)
  • Here we describe the protocols of using the chip to evaluate DNA damage caused by known genotoxic agents and the cellular repair response followed after exposure. (jove.com)
  • 8-oxo-dG-positive cells and the length of the comet tail increased within 1 h of exposure to H 2 O 2 . (biomedcentral.com)
  • In spite of a continual loss of ΔΨm, DNA fragmentation was reduced 2 h after exposure to H 2 O 2 . (biomedcentral.com)
  • With the increase of ozone exposure concentration, the tail rate of comets was increased gradually, and there was a significant dose-effect relationship. (bvsalud.org)
  • [email protected]#Acute exposure of ozone at low concentrations (0.12 ppm) could lead to DNA damage in the pulmonary cells of rats, while no significant chromosome damage was found even in the group with ozone concentration reached to 4 ppm. (bvsalud.org)
  • [email protected]#To investigate the vascular damage effects and possible mechanism of acute exposure to ozone (O) in male Wistar rats. (bvsalud.org)
  • C) Immunoprecipitation (IP) assays were performed in unperturbed cells or cells recovering from exposure to pulses of HU (0.2 mM, 24 h) at the indicated times after release using an anti-RPA32 antibody. (asm.org)
  • Our results showed that exposure of cells to 5 Gy of XR irradiation alone led to considerable DNA damage, which was significantly reduced by post-irradiation exposure to SMFs. (cancer-colorectal.com)
  • X-ray repair cross complementing-3 (XRCC3) is involved in the homologous recombination pathway of DNA DSB-repair [ 10 ] and the role XRCC3 gene have been most extensively studied due to their influences in the individual sensitivity to radiation exposure and induction of DNA damage. (biomedcentral.com)
  • Winter PM2.5 increased the level of ROS at 2 h and DNA damage (8-oxodG, single- and double stand breaks) was detected after 3 h of exposure. (biomedcentral.com)
  • Our data demonstrate that Sly deficiency is responsible for the sperm DNA damage/chromatin packaging defects observed in mice with NPYq deletions and point to SLY proteins involvement in chromatin reprogramming during spermiogenesis, probably through their effect on the post-meiotic expression of spermiogenic genes. (biologists.org)
  • The formation of γ-H2AX foci is believed to promote effective repair by aiding in the accumulation of checkpoint adaptor proteins and the recruitment of DNA repair machinery such as Brca1 and 53BP1 to damage sites ( 33 , 35 , 54 ). (asm.org)
  • Cells are then lysed in an alkaline pH buffer to remove cellular proteins and allow for DNA unwinding. (bioreliance.com)
  • 1 2 3 Although proteins, lipids, and DNA are all vulnerable, DNA damage is considered to be fundamental in the age-related decline in cellular function. (arvojournals.org)
  • Alterations in gene expression, the redistribution of existing proteins, and the assembly of new protein complexes can be stimulated by a variety of DNA lesions and mismatched DNA base pairs. (jove.com)
  • A number of proteins have been identified that are involved during the DNA damage checkpoint response. (genetics.org)
  • After staining with a DNA-specific fluorescent dye such as ethidium bromide or propidium iodide, the gel is read for amount of fluorescence in head and tail and length of tail. (sigmaaldrich.com)
  • The slides are then neutralised to pH 7, stained with a DNA-specific fluorescent stain and analysed using a microscope with an attached CCD (charge-coupled device - essentially a digital camera) that is connected to a computer with image analysis software. (scribd.com)
  • After rinsing the plates, DNA is marked with a fluorescent dye (e.g. ethidium, bromure, and acridine). (adcis.net)
  • Briefly, cells are embedded in agarose, lysed and submitted to an electric field, before the staining step with a fluorescent DNA binding dye. (bio-protocol.org)
  • Scoring was performed on a platform consisting of a Nikon 50i fluorescent microscope (Nikon, USA) and Comet IV digital imaging software ( Perceptive Instruments , Wiltshire, UK). (scorecomets.com)
  • In this report we demonstrate that Spy1 expression in mammalian cells enhances survival under conditions of genotoxic damage, including treatments of CPT, cisplatin, and HU. (aacrjournals.org)
  • In the present study, we investigated the effects of these flavonoids as antioxidants in chemical systems and plasmid DNA and radioprotectants in bacterial and mammalian cells. (scholarena.co)
  • The tails were evaluated through Comet Assay IV analysis software ( Perceptive Instruments Ltd ) and Camera AVT smart view for, Fire Package version 1.6.2) to measure the total image fluorescence intensity, head intensity, tail intensity, head length and tail length. (scorecomets.com)
  • The accumulation of DNA damage and the cellular response to DNA damage stressors was more pronounced in post-mitotic mFUS motoneurons than in dividing hiPSCs suggesting that mFUS motoneurons accumulate foci of DNA damage, which in turn might be directly linked to neurodegeneration. (frontiersin.org)
  • Consistent with Spy1 having a role in the DNA damage response, endogenous Spy1 protein levels are up-regulated in response to DNA damage induced by camptothecin, cisplatin, or hydroxyurea. (aacrjournals.org)
  • Analysis of subnuclear localization of GFP-tagged BRCA1 demonstrated that mutation of the PP1-binding motif affected BRCA1 redistribution in response to DNA damage. (ijbs.com)
  • BRCA1 is also phosphorylated by checkpoint kinases, including hCds1/Chk2, ATM, and ATR, in response to DNA damage induced by ionizing radiation (IR), UV or chemicals. (ijbs.com)
  • Serum lymphocyte DNA damage and total oxidant status (TOS), total antioxidant status (TAS) and oxidative stress index (OSI) were studied as oxidative stress parameters. (alliedacademies.org)
  • Serum lymphocyte DNA damage levels were 14.97 ± 12.38 AU (arbitrary units) in Group 1 (before analgesia), 13.94 ± 9.28 AU in Group 2 (2 h after analgesia), and 10.57 ± 8.73 AU in Group 3 (12 h after analgesia). (alliedacademies.org)
  • Serum concentrations of ferritin and soluble transferrin receptor were assayed by ELISA. (qxmd.com)
  • While well characterized in vitro, little is known about the time-course of DNA damage response in tumors sampled from individual patients. (mdpi.com)
  • Changes in gene expression induced by miR-183 in HTM cells were evaluated by gene array analysis, confirmed by quantitative-PCR (Q-PCR), and analyzed by MetaCore pathway analysis. (arvojournals.org)
  • In this study, we analyzed pathophysiological alterations associated with ALS related FUS mutations (mFUS) in human induced pluripotent stem cells (hiPSCs) and hiPSC derived motoneurons. (frontiersin.org)
  • DNA damage reflecting certain genotoxic exposures), and early biological effects (e.g., deleterious mutations). (aacrjournals.org)
  • Also, we obtained high success rates in determining the number of CAG repeats in the androgen receptor gene, characterizing tetranucleotide microsatellites in six gene loci, and screening for mutations in the BRCA1/2 genes in a subset of phenol-chloroform DNA extracts. (aacrjournals.org)
  • Brierley EJ, Johnson MA, Lightowlers RN, James OF, Turnbull DM (1998) Role of mitochondrial DNA mutations in human aging: implications for the central nervous system and muscle. (springer.com)
  • Somatic mutations and rearrangements in DNA molecule can lead to development of many degenerative disorders including atherosclerosis, autoimmune diseases, Alzheimer's disease, certain types of diabetes, and aging [ 5 - 9 ]. (intechopen.com)
  • The alteration can have direct or indirect effects on the DNA: the induction of mutations, mistimed event activation, and direct DNA damage leading to mutations. (wikipedia.org)
  • Samples of blood were collected from the females before copulation, after the 15th day of conception and on the 21st day after the lactation period, for the glycaemic and lipid profiles as well as comet assay and micronucleus (MN) test. (bvsalud.org)
  • In the gestation and lactation period, the two fructose doses tested showed DNA damage as observed in the comet assay, which is associated with an increase in dietary intake, body weight, lipid profile and fasting glycaemia in females. (bvsalud.org)
  • Numerous studies have elucidated that populations persistently exposed to As had a significant induction of a strong oxidative stress with generation of free radicals in cells that, in turn, mediates DNA damage as well as lipid peroxidation and decreased glutathione levels. (ijpsdr.com)
  • Sly deficiency has been shown to be the underlying cause of the sperm head anomalies and infertility associated with NPYq gene loss, but it was not known whether it recapitulates sperm DNA damage phenotype. (biologists.org)
  • In a case-control study, the baseline, benzo[a]pyrene diol epoxide (BPDE)-induced, and γ radiation-induced DNA damage were quantified by the Olive tail moment (TM) in PBLs from 172 Caucasian EAC patients and 154 healthy controls who were frequency matched on age and gender. (omicsonline.org)
  • EAC patients tended to have higher DNA damage than controls, as measured by baseline, net BPDEand net γ radiation-induced TM, but the difference was statistically significant only for net BPDE-induced DNA damage (0.88 ± 0.94 vs. 0.62 ± 0.77, P=0.031). (omicsonline.org)
  • Using the 75th percentile TM in the controls as cutoff point, we found that high levels of net BPDE- and γ radiation-induced DNA damage were associated with significantly increased risks of EAC, with adjusted ORs of 2.15 (95% CI, 1.13-4.10) and 2.27 (95% CI, 1.24-4.16), respectively. (omicsonline.org)
  • Our goal was to use a battery of different assays that complemented each other to further understand the role of Ot and Vc in protecting against chemical- and radiation-induced damage. (scholarena.co)
  • In this study, 2'-deoxyguanosine (dG) was exposed to N-(2,6-dimethylphenyl)hydroxylamine (2,6-DMPHA) and N-phenylhydroxylamine (PHA) at pH 7.4 without enzymes and analyzed by liquid chromatography high resolution mass spectrometry (LC-HRMS). (bvsalud.org)
  • The activities of aconitase and fumarase enzymes, which have key roles in energy metabolism, were analyzed in P. sativum cultures after treatment with NIC or NIA. (diva-portal.org)