scholarly journals Antigenotoxic Effect ofTrametesspp. Extracts against DNA Damage on Human Peripheral White Blood Cells

2015 ◽  
Vol 2015 ◽  
pp. 1-10 ◽  
Author(s):  
Aleksandar Knežević ◽  
Lada Živković ◽  
Mirjana Stajić ◽  
Jelena Vukojević ◽  
Ivan Milovanović ◽  
...  
Keyword(s):  
Dna Damage ◽  
Blood Cells ◽  
Conventional Medicine ◽  
White Blood Cells ◽  
Dpph Assay ◽  
Strand Breaks ◽  
Dna Migration ◽  
Dna Strand ◽  
Peripheral White Blood Cells ◽  
Antigenotoxic Effect

Trametesspecies have been used for thousands of years in traditional and conventional medicine for the treatment of various types of diseases. The goal was to evaluate possible antigenotoxic effects of mycelium and basidiocarp extracts of selectedTrametesspecies and to assess dependence on their antioxidant potential.Trametes versicolor, T. hirsuta, andT. gibbosawere the species studied. Antigenotoxic potentials of extracts were assessed on human peripheral white blood cells with basidiocarp and mycelium extracts of the species. The alkaline comet test was used for detection of DNA strand breaks and alkali-labile sites, as well as the extent of DNA migration. DPPH assay was used to estimate antioxidative properties of extracts. Fruiting body extracts ofT. versicolorandT. gibbosaas well asT. hirsutaextracts, except that at 20.0 mg/mL, were not genotoxic agents.T. versicolorextract had at 5.0 mg/mL the greatest antigenotoxic effect in both pre- and posttreatment of leukocytes. The mycelium extracts of the three species had no genotoxic activity and significant antigenotoxic effect against H2O2-induced DNA damage, both in pre- and posttreatment. The results suggest that extracts of these three species could be considered as strong antigenotoxic agents able to stimulate genoprotective response of cells.

scholarly journals DNA damage in human whole blood caused by radiopharmaceuticals evaluated by the comet assay

Mutagenesis ◽  
2019 ◽  
Vol 34 (3) ◽  
pp. 239-244 ◽  
Author(s):  
Heinz H Schmeiser ◽  
Karl-Rudolf Muehlbauer ◽  
Walter Mier ◽  
Ann-Christin Baranski ◽  
Oliver Neels ◽  
...  
Keyword(s):  
Dna Damage ◽  
Comet Assay ◽  
Dna Strand Breaks ◽  
Strand Breaks ◽  
Human Whole Blood ◽  
Time Points ◽  
Dna Migration ◽  
Activity Concentrations ◽  
Positron Emitters ◽  
Dna Strand

Abstract Radiopharmaceuticals used for diagnosis or therapy induce DNA strand breaks, which may be detectable by single-cell gel electrophoresis (called comet assay). Blood was taken from patients before and at different time points after treatment with radiopharmaceuticals; blood cells were investigated by the comet assay using the percentage of DNA in the tail as the critical parameter. Whereas [225Ac]Ac-prostate-specific membrane antigen (PSMA)-617 alpha therapy showed no difference relative to the blood sample taken before treatment, beta therapy with [177Lu]Lu-PSMA-617 3 h post-injection revealed a small but significant increase in DNA strand breaks. In blood of patients who underwent positron emission tomography (PET) with either [18F]2-fluor-2-deoxy-D-glucose (FDG) or [68Ga]Ga-PSMA-11, an increase of DNA migration determined by the comet assay was not found when analysed at different time points (2–70 min) after intravenous tracer injection. Human whole blood was incubated with the targeted clinically relevant therapeutic radiopharmaceuticals [225Ac]Ac-PSMA-617, [177Lu]Lu-PSMA-617 and [90Y]Y-DOTA(0)-Phe(1)-Tyr(3)-octreotide (DOTA-TOC) at different activity concentrations (kBq/ml) for 5 days and then analysed by the comet assay. DNA damage increased with higher concentrations of all radiolabeled compounds tested. [177Lu]Lu-PSMA-617 caused higher blood cell radiotoxicity than equal activity concentrations of [90Y]Y-DOTA-TOC. Likewise, whole human blood was exposed to the positron emitters [18F]FDG and [68Ga]Ga-PSMA-11 in vitro for 24 h with activity concentrations ranging between 5 and 40 MBq/ml. The same activity concentration dependent elevated DNA migration was observed for both compounds although decay energies are different. This study demonstrated that the amount of DNA damage detected by the comet assay in whole human blood is similar among different positron emitters and divergent by a factor of 200 between alpha particles and beta radiation.

scholarly journals Atmospheric pollution in Kosovo is associated with increased DNA damage in the human population

Biomonitoring ◽  
2016 ◽  
Vol 2 (1) ◽  
Author(s):  
Avdulla J. Alija ◽  
Fisnik Asllani ◽  
Ismet D. Bajraktari ◽  
Andrew Richard Collins ◽  
Shemsedin Dreshaj ◽  
...  
Keyword(s):  
Dna Damage ◽  
Blood Cells ◽  
Power Plants ◽  
White Blood Cells ◽  
Control Site ◽  
Alkaline Comet Assay ◽  
Potential Threat ◽  
Strand Breaks ◽  
Unpolluted Area ◽  
Polluted Sites

Abstract: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. White blood cells were isolated, and DNA damage was analyzed by the alkaline comet assay. Significantly higher levels of DNA damage (strand breaks) were found in white blood cells from subjects living in the polluted areas compared with residents of the unpolluted city, indicating a potential threat to human health.

Measurement of DNA damage with the comet assay in high-prevalence diseases: current status and future directions

Mutagenesis ◽  
2019 ◽  
Author(s):  
Peter Møller ◽  
Helga Stopper ◽  
Andrew R Collins
Keyword(s):  
Dna Damage ◽  
Comet Assay ◽  
White Blood Cells ◽  
Dna Strand Breaks ◽  
Chronic Obstructive ◽  
Case Control Studies ◽  
Obstructive Pulmonary Disease ◽  
Strand Breaks ◽  
Artery Disease ◽  
Dna Strand

Abstract The comet assay is widely used in studies on genotoxicity testing, human biomonitoring and clinical studies. The simple version of the assay detects a mixture of DNA strand breaks and alkali-labile sites; these lesions are typically described as DNA strand breaks to distinguish them from oxidatively damaged DNA that are measured with the enzyme-modified comet assay. This review assesses the association between high-prevalence diseases in high-income countries and DNA damage measured with the comet assay in humans. The majority of case–control studies have assessed genotoxicity in white blood cells. Patients with coronary artery disease, diabetes, kidney disease, chronic obstructive pulmonary disease and Alzheimer’s disease have on average 2-fold higher levels of DNA strand breaks compared with healthy controls. Patients with coronary artery disease, diabetes, kidney disease and chronic obstructive pulmonary disease also have 2- to 3-fold higher levels of oxidatively damaged DNA in white blood cells than controls, although there is not a clear difference in DNA damage levels between the different diseases. Case–control studies have shown elevated levels of DNA strand breaks in patients with breast cancer, whereas there are only few studies on colorectal and lung cancers. At present, it is not possible to assess if these neoplastic diseases are associated with a different level of DNA damage compared with non-neoplastic diseases.

scholarly journals Repair of DNA Strand Breaks by the Overlapping Functions of Lesion-Specific and Non-Lesion-Specific DNA 3′ Phosphatases

2001 ◽  
Vol 21 (21) ◽  
pp. 7191-7198 ◽  
Author(s):  
John R. Vance ◽  
Thomas E. Wilson
Keyword(s):  
Dna Damage ◽  
Oxidative Dna Damage ◽  
Synthetic Lethality ◽  
Dna Strand Breaks ◽  
Triple Mutant ◽  
Phosphatase Domain ◽  
Strand Breaks ◽  
Alternative Pathways ◽  
Processing Enzymes ◽  
Dna Strand

ABSTRACT In Saccharomyces cerevisiae, the apurinic/apyrimidinic (AP) endonucleases Apn1 and Apn2 act as alternative pathways for the removal of various 3′-terminal blocking lesions from DNA strand breaks and in the repair of abasic sites, which both result from oxidative DNA damage. Here we demonstrate that Tpp1, a homologue of the 3′ phosphatase domain of polynucleotide kinase, is a third member of this group of redundant 3′ processing enzymes. Unlike Apn1 and Apn2, Tpp1 is specific for the removal of 3′ phosphates at strand breaks and does not possess more general 3′ phosphodiesterase, exonuclease, or AP endonuclease activities. Deletion ofTPP1 in an apn1 apn2 mutant background dramatically increased the sensitivity of the double mutant to DNA damage caused by H2O2 and bleomycin but not to damage caused by methyl methanesulfonate. The triple mutant was also deficient in the repair of 3′ phosphate lesions left by Tdp1-mediated cleavage of camptothecin-stabilized Top1-DNA covalent complexes. Finally, the tpp1 apn1 apn2 triple mutation displayed synthetic lethality in combination with rad52, possibly implicating postreplication repair in the removal of unrepaired 3′-terminal lesions resulting from endogenous damage. Taken together, these results demonstrate a clear role for the lesion-specific enzyme, Tpp1, in the repair of a subset of DNA strand breaks.

scholarly journals Engineered Nanoparticles Induce DNA Damage in Primary Human Skin Cells, Even at Low Doses

Nano LIFE ◽  
2014 ◽  
Vol 04 (01) ◽  
pp. 1440001 ◽  
Author(s):  
Amelia A. Romoser ◽  
Michael F. Criscitiello ◽  
Christie M. Sayes
Keyword(s):  
Dna Damage ◽  
Stress Protein ◽  
Dermal Fibroblasts ◽  
Engineered Nanoparticles ◽  
Ros Generation ◽  
Heme Oxygenase 1 ◽  
Dna Double Strand Breaks ◽  
Biological Reduction ◽  
Strand Breaks ◽  
Dna Strand

It is well documented that various particulate matter — either incidental or engineered — are known to generate reactive oxygen species (ROS) in living cells. In circumstances where these reactive species are generated, antioxidant production is often increased. This balance in the biological reduction/oxidation (a.k.a. redox) state within the cell has not been thoroughly studied in exposures involving engineered nanoparticles. However, nanoparticle exposure has been postulated to induce a DNA damage cascade. In this study, we examined primary human dermal fibroblasts (HDF) exposed to three different, but commonly used engineered nanoparticles (i.e., cerium dioxide ( CeO 2), titanium dioxide ( TiO 2) and zinc oxide ( ZnO )) in an attempt to determine the potential DNA damaging effects through the analysis of ROS generation, relevant protein upregulation response and single and double DNA strand breaks. Cell death was most elevated with exposure to ZnO , followed by TiO 2 and CeO 2. ROS generation was measured at 1 h, 6 h and 24 h after exposure to particles via a cell-based DCFH-DA (2′, 7′-dichlorfluorescein-diacetate) assay and indicated that ZnO generated the most significant amount of ROS. ZnO also caused upregulation of oxidative stress protein, heme oxygenase-1 and phosphorylation of p38; whereas CeO 2 caused upregulation of superoxide dismutase. Results from the comet assay indicated that ZnO triggered significant DNA damage in cells at relatively low dosing concentrations (20 ppm). Immunocytochemistry with ZnO -treated cells revealed notable DNA double strand breaks evidenced by a marked increase in the presence of γ-H2AX foci. This finding was also indicated by western blot, as well as cell cycle arrest by the phosphorylation of cyclin-dependent kinase 1. These data suggest that the three particle-types induce different degrees of DNA damage. And, of the three particle-types tested, exposure to ZnO nanoparticles may cause the most significant DNA damage.

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