Evaluation of DNA damage in rat lymphocytes exposed to tulathromycin in vitro

Tulathromycin is a relatively new semi-synthetic macrolide antibiotic, a member of the triamilide group, approved primarly for the treatment of respiratory diseases in cattle and swine. Various genotoxicological studies indicated that tulathromycin is not genotoxic, but no available published data originate from the single-cell gel electrophoresis (Comet) assay. Therefore, the objective of this study was to examine whether it can induce primary DNA damage using in vitro Comet assay in isolated rat lymphocytes. Lymphocytes were treated with a broad spectrum of tulathromycin concentrations (from 1 to 100 ? M) and co-treatment with an antioxidant, catalase (100 IU/mL and 500 IU/mL) was performed. The highest concentrations of tulathromycin (50 and 100 ? M) caused significant increase of DNA damage in rat lymphocytes and catalase did not significantly reduce the DNA-damaging effect of tulathromycin. The results of this study indicate that tulathromycin induces genotoxic effects at high concentrations, that catalase does not exert protective effect in this case.

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In vitro genotoxic effects of the insecticide deltamethrin in human peripheral blood leukocytes: DNA damage (‘comet’ assay) in relation to the induction of sister-chromatid exchanges and micronuclei

Toxicology ◽

10.1016/s0300-483x(98)00097-3 ◽

1998 ◽

Vol 130 (2-3) ◽

pp. 129-139 ◽

Cited By ~ 83

Author(s):

Milena Villarini ◽

Massimo Moretti ◽

Rossana Pasquini ◽

Giuseppina Scassellati-Sforzolini ◽

Cristina Fatigoni ◽

...

Keyword(s):

Dna Damage ◽

Comet Assay ◽

Sister Chromatid ◽

Human Peripheral Blood ◽

Sister Chromatid Exchanges ◽

Peripheral Blood Leukocytes ◽

Genotoxic Effects ◽

Blood Leukocytes ◽

Chromatid Exchanges

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Genotoxic effects of nanosized and fine TiO2

Human & Experimental Toxicology ◽

10.1177/0960327109105163 ◽

2009 ◽

Vol 28 (6-7) ◽

pp. 339-352 ◽

Cited By ~ 146

Author(s):

GCM Falck ◽

HK Lindberg ◽

S. Suhonen ◽

M. Vippola ◽

E. Vanhala ◽

...

Keyword(s):

Dna Damage ◽

Comet Assay ◽

Micronucleus Test ◽

Genotoxic Effects ◽

Bronchial Epithelial ◽

Dose Dependent ◽

Dose Dependent Effect ◽

Fine Tio2 ◽

In Vitro Genotoxicity

The in-vitro genotoxicity of nanosized TiO2 rutile and anatase was assessed in comparison with fine TiO2 rutile in human bronchial epithelial BEAS 2B cells using the single-cell gel electrophoresis (comet) assay and the cytokinesis-block micronucleus test. BEAS 2B cells were exposed to eight doses (1—100 μg/cm2) of titanium(IV) oxide nanosized rutile (>95%, <5% amorphous SiO2 coating; 10 × 40 nm), nanosized anatase (99.7%; <25 nm), or fine rutile (99.9%; <5 μm) for 24, 48, and 72 h. Fine rutile reduced cell viability at lower doses than nanosized anatase, which was more cytotoxic than nanosized rutile. In the comet assay, nanosized anatase and fine rutile induced DNA damage at several doses with all treatment times. Dose-dependent effects were seen after the 48- and 72-h treatments with nanosized anatase and after the 24-, 48- (in one out of two experiments), and 72-h treatments (one experiment) with fine rutile. The lowest doses inducing DNA damage were 1 μg/cm2 for fine rutile and 10 μg/cm 2 for nanosized anatase. Nanosized rutile showed a significant induction in DNA damage only at 80 μg/cm2 in the 24-h treatment and at 80 and 100 μg/ cm2 in the 72-h treatment (with a dose-dependent effect). Only nanosized anatase could elevate the frequency of micronucleated BEAS 2B cells, producing a significant increase at 10 and 60 μg/cm 2 after the 72-h treatment (no dose-dependency). At increasing doses of all the particles, MN analysis became difficult due to the presence of TiO2 on the microscopic slides. In conclusion, our studies in human bronchial epithelial BEAS 2B cells showed that uncoated nanosized anatase TiO2 and fine rutile TiO2 are more efficient than SiO 2-coated nanosized rutile TiO2 in inducing DNA damage, whereas only nanosized anatase is able to slightly induce micronuclei.

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The Investigation of DNA Damage Induced by Adrenaline in Human Lymphocytes in Vitro/Ispitivanja Oštećenja DNK Izazvanih Adrenalinom U Limfocitima Čoveka in Vitro

Acta veterinaria ◽

10.2478/acve-2014-0027 ◽

2014 ◽

Vol 64 (3) ◽

pp. 281-292 ◽

Cited By ~ 5

Author(s):

Radaković Milena ◽

Đelić Ninoslav ◽

Stevanović Jevrosima ◽

Anđelković Marko ◽

Kolarević Stoimir ◽

...

Keyword(s):

Reactive Oxygen Species ◽

Dna Damage ◽

Human Lymphocytes ◽

Reactive Oxygen ◽

Potential Contribution ◽

Oxygen Species ◽

Genotoxic Effects ◽

Isolated Human Lymphocytes ◽

Primary Dna Damage

Abstract Adrenaline is a neurotransmitter and hormone that plays an important role in physiological regulatory mechanisms. The objective of this study was to assess primary DNA damage in isolated human lymphocytes exposed to adrenaline using the in vitro comet assay. Dose-response of human lymphocytes was determined at concentration range of adrenaline from 0.01 μM to 300 μM for various treatment times (1h, 2h, 4h and 24h). The obtained results showed that adrenaline induced DNA damage at concentration range from 5 μM to 300 μM after 1h, 2h and 4h of treatment. The slightest DNA damage was observed after 24 h of adrenaline treatment - only the highest concentrations of adrenaline (150 μM and 300 μM) caused increased level of DNA damage. In order to evaluate the potential contribution of reactive oxygen species (ROS) in adrenaline-induced DNA damage we used antioxidants catalase (100 IU/mL and 500 IU/mL) and quercetin (100 μM and 500 μM). Co-treatment of lymphocytes with adrenaline (300 μM) and antioxidants for 1 h, significantly reduced the quantity of DNA in the comet tails. Therefore, it can be concluded that adrenaline exhibits genotoxic effects mainly through induction of reactive oxygen species and that some of the DNA damage is repaired during the first four hours following the treatment with adrenaline.

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In vitro assessment of the genotoxicity of ethyl paraoxon in newborns and adults

Human & Experimental Toxicology ◽

10.1191/0960327105ht534oa ◽

2005 ◽

Vol 24 (6) ◽

pp. 319-324 ◽

Cited By ~ 8

Author(s):

K Islas-González ◽

C González-Horta ◽

B Sánchez-Ramírez ◽

E Reyes-Aragón ◽

M Levario-Carrillo

Keyword(s):

Comet Assay ◽

Study Groups ◽

Major Effect ◽

Blood Samples ◽

Dna Migration ◽

In Vitro Assessment ◽

High Concentrations ◽

And Control ◽

Primary Dna Damage

This in vitro experiment measured the genotoxic effects of ethyl paraoxon, the active metabolite of ethyl parathion. To assess genotoxicity, we used the micronuclei (MN) technique by blocking cytokinesis, and the ‘comet’ assay. We cultured peripheral blood samples from healthy adults and umbilical cord blood samples from four clinically healthy newborns to identify the frequency of MN. After 48 hours, we added the following ethyl paraoxon concentrations to the cultures: 0.0, 0.075, 0.100, 0.160, and 0.200 μg/mL. For the comet assay, following Singh's technique, we treated the blood samples for 2 hours with similar doses of the metabolite. The comet assay results, at a concentration of 0.075 μg/mL, showed that ethyl paraoxon causes a greater DNA migration that followed a dose-response pattern, a greater intensity being observed in lymphocytes from newborns. A comparison of the treatment and control groups indicated that only the 0.200 μg/mL concentration produced a slight increase in MN. In conclusion, our study identified primary DNA damage due to ethyl paraoxon, with a major effect on newborn lymphocytes, as well as an effect on the frequency of MN in the study groups at high concentrations only.

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Assessment of genotoxic effects in one-and two-cell mouse embryos in vivo and in vitro using comet assay

Nauchno-prakticheskii zhurnal «Medicinskaia genetika» ◽

10.25557/2073-7998.2020.09.77-78 ◽

2020 ◽

pp. 77-78

Author(s):

К.Л. Плигина ◽

А.К. Жанатаев ◽

Е.А. Анисина ◽

Н.О. Даугель-Дауге ◽

А.Д. Дурнев

Keyword(s):

Dna Damage ◽

Comet Assay ◽

Mitomycin C ◽

Mouse Embryos ◽

Methyl Methanesulfonate ◽

Genotoxic Effects

Разработана методология оценки первичных повреждений ДНК в одно- и двухклеточных зародышах мышей методом ДНК-комет. Применимость разработанной методологии для оценки генотоксичности в зародышевых клетках in vivo и in vitro подтверждена в экспериментах с модельными генотоксикантами метилметансульфонатом, диоксидином, этопозидом и митомицином С. A methodology for evaluating DNA damage in one - and two-cell mouse embryos using the comet assay has been developed. The applicability of the developed methodology for assessing genotoxicity in vivo and in vitro was confirmed in experiments with model genotoxicants - methyl methanesulfonate, dioxidine, etoposide and mitomycin C.

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