Somatic Mutation And Recombination Test Research Articles

Investigation of the Genotoxic Effect of Fluoxetine Hydrochloride in Drosophila melanogaster

This study aimed to determine the potential genotoxic effect of fluoxetine hydrochloride (FLX-HCl), an antidepressant commonly used for treating depression, using Somatic Mutation and Recombination Test (SMART). Third-¬instar Drosophila melanogaster larvae transheterozygous for the mutations multiple wing hair (mwh) and flare (flr3) were chronically fed in a medium containing different concentrations of FLX-HCl (0.1, 0.5, 1, and 2 mg/mL) in the experimental group. Distilled water, 0.1 mM ethyl methane sulfonate (EMS), and 2% dimethyl sulfoxide (DMSO) were used in negative, positive, and solvent control groups, respectively. The survival percentages were calculated by determining the number of individuals surviving when the larvae completed their development in the experimental and control groups. In all application groups, the wings of 40 individuals with both normal and serrate wing phenotypes were examined under a microscope, and genetic changes were evaluated by counting the mutant clones in the wings. The data obtained show that 1 and 2 mg/mL concentrations of FLX-HCl caused toxic effects in D. melanogaster individuals. Additionally, FLX-HCl showed a negative genotoxic effect at 0.1 mg/mL concentration, insignificant at 0.5 mg/mL concentration, and positive at 1 and 2 mg/mL concentrations in terms of total mutation evaluation and clone induction frequency in D. melanogaster individuals.

Antigenotoxic and Lifespan-Promoting Effects of Flavoured Kombuchas on Drosophila melanogaster

Research background. Kombucha is a fermented beverage with several health benefits; however, to improve its antioxidant activity, new raw materials consisting of hop, madimak, and hawthorn were included in the present study. Experimental approach. The Drosophila Somatic Mutation and Recombination Test (SMART) was used to evaluate the antigenotoxic potential of black tea kombucha and three-flavoured kombuchas (hop, madimak, and hawthorn) against H2O2- and K2Cr2O7-induced genotoxicity. Furthermore, a lifespan assay was performed to assess the effects of kombuchas on Drosophila longevity. Results and conclusions. According to the results obtained from the SMART assay, hop-flavoured kombucha attenuated genotoxicity induced by H2O2, and madimak-flavoured kombucha reduced genotoxicity induced by H2O2 and K2Cr2O7. Black tea kombucha and hop-flavoured kombucha prolonged the lifespan of Drosophila after H2O2 and K2Cr2O7 exposure. Novelty and scientific contribution. Hop-flavoured kombucha is a promising antioxidant that protects the genome and extends the lifespan of Drosophila. This study sheds light on novel beverages that can combat aging and protect against genotoxicity.

EFFECT OF BITTER MELON AGAINST CISPLATIN AND VALPROIC ACID-INDUCED GENOTOXICITY IN Drosophila melanogaster Meigen

In human body, which is exposed to number of chemicals and physical agents in daily life, malfunctions may occur from time to time in detoxification mechanisms that will prevent the harmful effects of genotoxic agents, whose negative effects cannot be eliminated. In this study, the genoprotective effects of bitter melon (Momordica charantia L.) seed and fruit extracts against the genotoxic potential of the antineoplastic agent cisplatin and the antiepileptic drug valproic acid in 72±4 hours old transheterozygous larvae of the fruit fly Drosophila melanogaster Meigen were determined by the Somatic Mutation and Recombination Test (SMART). The results showed that the mutation frequencies approached the negative control values by inhibiting the formation of chemical agent-induced mutant clones in all doses in the experimental groups in which plant extracts were applied at doses of 1.25, 2.5 ve 5 mg/mL, while the mutation frequency increased only in the groups that applied cisplatin and valproic acid. These results show that co-administration of cisplatin and valproic acid reduces the genotoxic effect of M. charantia.

Copper(II) Complex Containing 4-Fluorophenoxyacetic Acid Hydrazide and 1,10-Phenanthroline: A Prostate Cancer Cell-Selective and Low-Toxic Copper(II) Compound.

Prostate Cancer (PCa) is the second leading cause of cancer-related deaths among men worldwide. The treatment of advanced cases is based on chemotherapy, which lacks specificity and efficacy, due to severe side effects and resistance to the traditional drugs. Copper complexes have shown antitumoral efficacy and low toxicity, being considered a promising class of metal-based drugs for the treatment of malignant neoplasms. Thus, the present study aimed to evaluate the cellular effects of a copper(II) complex with 4-fluorophenoxyacetic acid hydrazide and 1,10-phenanthroline (1) on PCa cell lines, as well as the mutagenic/recombinogenic and anticarcinogenic potential of 1 in Drosophila melanogaster. PNT-2 (non-tumorigenic), LNCaP (hormone-responsive PCa) and PC-3 (androgen-independent PCa) cells were cultured, and cytotoxicity was assessed using the MTT assay. The expression levels of the proliferation markers Ki-67 and Cyclin D1 were analyzed by flow cytometry. Furthermore, the Somatic Mutation and Recombination Test (SMART) and the Epithelial Tumor Test (ETT) were performed. Complex 1 was selective to LNCaP cells, significantly reducing Ki-67 and Cyclin D1 expression levels. Sub-toxic concentrations of complex 1 were defined by the toxicity test in D. melanogaster, and no mutagenic/recombinogenic/carcinogenic effects were observed. Anticarcinogenic potential was observed in D. melanogaster, suggesting modulating activity of the complex 1 against Doxorubicin, a drug used as control by its carcinogenic properties. Therefore, complex 1 is a possible starting point for the development of new antitumor agents for the treatment of PCa.

Evaluation of Mutagenicity and Anti-Mutagenicity of Various Bean Milks Using Drosophila with High Bioactivation.

The consumption of a nutritious diet including phytochemicals can minimize mutations as the primary cause of carcinogenesis. Bean consumption supplies calories, minerals and phytochemicals but their anti-mutagenic properties in vivo remain little understood. Hence, the present study aimed to study the mutagenicity and anti-mutagenic properties of five bean milks using the somatic mutation and recombination test (SMART) involving Drosophila with high bioactivation. Milk derived from five bean varieties, namely black bean (Phaseolus vulgaris), red kidney bean (Phaseolus vulgaris), mung bean (Phaseolus aureus), peanut (Arachis hypogaea) and soybean (Glycine max) did not induce DNA mutations in Drosophila with high bioactivation, indicating their genome-safe properties. All bean milks showed anti-mutagenicity against the food-derived mutagen, urethane, in vivo with different degrees of inhibition. In the co-administration study, larvae were treated with each bean milk together with urethane. Soybean milk showed the highest anti-mutagenicity at 27.75%; peanut milk exhibited the lowest at 7.51%. In the pre-feeding study, the larvae received each bean milk followed by urethane. Soybean milk exhibited the highest anti-mutagenic potential, followed by red kidney bean and black bean milks. Total phenolic and antioxidant data revealed that the anti-mutagenicity of both red kidney bean milk and black bean milk might be derived from their phenolic or antioxidant properties; other phytochemicals may contribute to the high anti-mutagenicity observed in soybean milk. Further investigations on the anti-mutagenicity of bean milks against other dietary mutagens are required to develop bean-based products with potent anti-mutagenic properties.

Determination of the genotoxic effects of various dioxins by <em>Drosophila </em>wing spot test

Dioxins, furans and other polychlorinated biphenyls (coplanar PCBs) are three structural and toxicologically related families of compounds classified as the most toxic synthetic chemical. In this study, we investigated possible genotoxic effects of polychlorinated dibenzo-p-dioxins (PCDDs), man-made environmental contaminant, in Drosophila melanogaster by somatic mutation and recombination test (SMART). In this, we observe the chemical effects on wing phenotype of the transheterozygote flies carrying marker gene. Lethal doses of 2,3,7,8-TCDD, 1,2,3,7,8-PeCDD, 1,2,3,7,8,9-HXCDD, and 1,2,3,4,6,7,8,9-OCDD were determined. Doses of 1×10-7, 2.5×10-7, 5×10-7, and 10×10-7 μg/mL of PCDDs were used. In addition, the observed mutations were classified according to the size and the type of the mutations per wing. Results revealed no significant genotoxic effect of any of the dioxins tested. According to the mechanisms involved in the antigenotoxicity of PCDDs, it is suggested that the observed effects can be linked to the differences in the aryl hydrocarbon receptor (AHR) amino acid sequences in the gene protein of D.melanogaster

Publication trends of somatic mutation and recombination tests research: a bibliometric analysis (1984‒2020).

Human exposure to pollutants has been on the rise. Thus, researchers have been focused on understanding the effect of these compounds on human health, especially on the genetic information by using various tests, among them the somatic mutation and recombination tests (SMARTs). It is a sensitive and accurate method applicable to genotoxicity analysis. Here, a comprehensive bibliometric analysis of SMART assays in genotoxicity studies was performed to assess publication trends of this field. Data were extracted from the Web of Science database and analyzed by the bibliometric tools HistCite, Biblioshiny (RStudio), VOSViewer, and CiteSpace. Results have shown an increase in the last 10 years in terms of publication. A total of 392 records were published in 96 sources mainly from Brazil, Spain, and Turkey. Research collaboration networks between countries and authors were performed. Based on document co-citation, five large research clusters were identified and analyzed. The youngest research frontier emphasized on nanoparticles. With this study, how research trends evolve over years was demonstrated. Thus, international collaboration could be enhanced, and a promising field could be developed.

Study of the modulator effect of oil chia (Salvia hispanica L.) associated with benzo(a)pyrene and doxorubicin hydrochloride

Chia (Salvia hispanica L.) is a Mexican plant belonging to the Lamiaceae family, representing one of the four main seeds cultivated by pre-Columbian peoples, mainly the Aztecs who used it as a medicine and nutritional component. Currently, it has been rediscovered, they are rich in polyunsaturated fatty acids, with antioxidant activity and essential compounds for organism maintenance. Various benefits are attributed to this plant including the reduction of cardiovascular problems, psoriasis, depression, Alzheimer’s, diabetes, arthritis and cancer. Considering the widespread use of chia oil (OC) in today’s society, it was justified to investigate the modulating effect of this oil when combined with benzo(a)pyrene and doxorubicin hydrochloride, using the SMART (Somatic Mutation and Recombination Test), a test able to detect mutation and somatic recombination activities, with loss of heterozygosis, which can be observed in the forms of different trichomes present in the wings of Drosophila melanogaster. The results obtained indicate that chia oil reduced the frequency of mutations by 97% in the number of spots induced by benzo(a)pyrene (BaP) and doxorubicin hydrochloride (DRX), showing an antigenotoxic effect.

In vivo and in silico approaches to assess surface water genotoxicity from Tocantins River, in the cities of Porto Nacional and Palmas, Brazil

The main environmental problem in urban areas, especially in Brazil, is the discharge of untreated sewage. The in vivo Drosophila melanogaster Somatic Mutation and Recombination Test (SMART) was used to assess the genotoxicity of surface waters from three different sites in the Tocantins River, Brazil. The in silico approach was used to search for known and predicted interactions between environmental chemicals found in our samples and Drosophila and human proteins. The genotoxicity tests were performed in standard (ST) and high bioactivation (HB) crosses with samples collected at two periods, the rainy and dry seasons. Mutant spot frequencies found in treatments with unprocessed water from the test sites were compared with the frequencies observed in negative controls. The collection points were represented as sites A, B and C along Tocantins River. Sites A and B are located in Porto Nacional City, whereas site C is located in Palmas City. Considering the rainy season collection, positive responses in the ST cross were observed for sites A and C (89.47% and 85% of recombination, respectively) and in the HB cross for sites A, B and C (88.24%, 84.21% and 82.35% of recombination, respectively). The positive results in the dry season were restricted to sites A and B (88.89% and 85.71% of recombination, respectively) in the HB cross. In accordance with in vivo and in silico results, we hypothesize that ribosomal proteins (RPs) in fruit fly and humans are depleted in cells exposed to heavy metal causing DNA damage and chromosome instability, increasing homologous recombination.

Genoprotective role of purslane methanol extract against somatic mutations induced by bifenthrin, a third generation prethyroid insecticide

In this study, in vitro and in vivo genotoxic effects of Bifenthrin (BIF), an important insecticide used in agricultural production, storage and processing, were investigated. The genoprotective properties of purslane (Portulaca oleracea L.) against the genotoxic effects of BIF were also determined by using the methanol (POmet) extract of this plant. In vivo experiments were performed with somatic mutation and recombination test (SMART) in Drosophila melanogaster. In in vitro studies, human peripheral blood cultures were prepared and different concentrations of BIF were applied to lymphocyte cells in accordance with the procedure of both the micronucleus (MN) and sister chromatid exchange (SCE) assay. The results obtained from all applied tests showed that BIF is genotoxic and induces chromosomal mutations. Later, another experiment was conducted and it was determined that the genotoxic effects of BIF were reduced with POmet (1:1 v/v). This result, which was observed in all in vivo and in vitro tests, shows that purslane plant is a potent radical scavenger. Due to the healing properties of POmet, gas chromatography-mass spectrometry (GS-MS) method was used to determine the components in its content. Some of the components found in the highest ratio in this extract are γ-sitosterol (21.86%), 13-docosenamide, (13.30%), palmitic acid (12.85%), stigmasteol (6.64%), campesterol (5.69%), linoleicacid (5.46%) and 2-methyl-1-hexadecanol (3.88%).

Human Hazard Assessment Using Drosophila Wing Spot Test as an Alternative In Vivo Model for Genotoxicity Testing-A Review.

Genomic instability, one of cancer’s hallmarks, is induced by genotoxins from endogenous and exogenous sources, including reactive oxygen species (ROS), diet, and environmental pollutants. A sensitive in vivo genotoxicity test is required for the identification of human hazards to reduce the potential health risk. The somatic mutation and recombination test (SMART) or wing spot test is a genotoxicity assay involving Drosophila melanogaster (fruit fly) as a classical, alternative human model. This review describes the principle of the SMART assay in conjunction with its advantages and disadvantages and discusses applications of the assay covering all segments of health-related industries, including food, dietary supplements, drug industries, pesticides, and herbicides, as well as nanoparticles. Chemopreventive strategies are outlined as a global health trend for the anti-genotoxicity of interesting herbal extract compounds determined by SMART assay. The successful application of Drosophila for high-throughput screening of mutagens is also discussed as a future perspective.

Synthesis, Characterization, and Optimization of Green Silver Nanoparticles Using Neopestalotiopsis clavispora and Evaluation of Its Antibacterial, Antibiofilm, and Genotoxic Effects

Abstract Silver nanoparticles (AgNPs) have been used in a variety of biomedical applications in the last two decades, including antimicrobial, anti-inflammatory, and anticancer treatments. The present study highlights the extracellular synthesis of silver nanoparticles AgNPs using Neopestalotiopsis clavispora MH244410.1 and its antibacterial, antibiofilm, and genotoxic properties. Locally isolated N. clavispora MH244410.1 was identified by Internal transcribed spacer (ITS) sequences of nuclear ribosomal DNA. Optimization of synthesized AgNPs was performed by using various parameters (pH (2, 4, 7, 9 and 12), temperature (25, 35 and 45 °C), and substrate concentration (0.05, 0.1, 0.15, 0.2 and 0.25 mM)). After 72 hours of incubation in dark conditions, the best condition for the biosynthesis of AgNPs was determined as 0.25 mM metal concentration at pH 12 and 35 °C. Fungal synthesized AgNPs were characterized via spectroscopic and microscopic techniques such as Fouirer Transform Infrared Spectrophotometer (FTIR), UV-Visible Spectroscopy, and Transmission Electron Microscopy (TEM). The average size of the AgNPs was determined less than 60 nm using the TEM and Zetasizer measurement system (measured in purity water suspension). The characteristic peak of AgNPs was observed at ~414 nm from UV-Vis results. Antibacterial and genotoxic activity of synthesized AgNPs (0.1, 1, and 10 ppm) were also determined by using the agar well diffusion method and in vivo Somatic Mutation and Recombination Test (SMART) in Drosophila melanogaster. AgNPs exhibited potential antimicrobial activity against all the tested bacteria (Bacillus subtilis, Staphylococcus aureus, and Pseudomonas aeruginosa) except Escherichia coli in a dose-dependent manner. AgNPs did not induce genotoxicity in the Drosophila SMART assay. 79.33, 65.47, and 41.95% inhibition of biofilms formed by P. aeruginosa were observed at 10, 1, and 0.1 ppm of AgNPs, respectively. The overall results indicate that N. clavispora MH244410.1 is a good candidate for novel applications in biomedical research.

Modulating effect of DL-kavain on the mutagenicity and carcinogenicity induced by doxorubicin in Drosophila melanogaster

ABSTRACT Kavain, kavalactone, present in Piper methysticum exhibits anticonvulsive, analgesic, anxiolytic, antiepileptic, antithrombotic, anti-inflammatory and antioxidant properties. Given its importance, the aim of the present study was to assess (1) the mutagenic and carcinogenicity of kavain administered alone and (2) the antimutagenic and anticarcinogenic potential when administered simultaneously with the chemotherapeutic drug doxorubicin (DXR) using the Somatic Mutation and Recombination Test (SMART) and Epithelial Tumor Test (ETT) using Drosophila melanogaster as a model system. Third-stage larvae from a standard (ST) and high metabolic bioactivation (HB) crosses were treated with different kavain concentrations (32, 64 or 128 μg/ml), alone or in conjunction with DXR (0.125 mg/ml). In ST descendants, kavain produced no significant mutagenic or recombinogenic effects. In the HB cross, mutagenic activity was observed at kavain concentrations of 64 and 128 μg/ml. In the DXR and kavain co-treatment, a modulating effect of the DXR-mediated mutagenic response dependent upon the concentration was detected in both crosses. In ETT, no marked carcinogenic or anticarcinogenic activity was noted for kavain. However, when kavain was combined with DXR synergistic induction of tumors by the chemotherapeutic drug occurred indicating that kavain enhanced the carcinogenic action of DXR.

Mutagenic and genotoxic activities of Phospholipase A2 Bothropstoxin-I from Bothrops jararacussu in Drosophila melanogaster and human cell lines

Phospholipase A2 Bothropstoxin-I (PLA2 BthTX-I) is a myotoxic Lys49-PLA2 from Bothrops jararacussu snake venom. In order to evaluate the DNA damage caused by BthTX-I, we used the Somatic Mutation and Recombination Test (SMART) in Drosophila melanogaster and Comet assay in HUVEC and DU-145 cells. For SMART, different concentrations of BthTX-I (6.72 to 430 μg/mL) were used and no significant changes in the survival rate were observed. Significant frequency of mutant spots was observed for the ST cross at the highest concentration of BthTX-I due to recombinogenic activity. In the HB cross, BthTX-I increased the number of mutant spots at intermediate concentrations, being 53.75 μg/mL highly mutagenic and 107.5 μg/mL predominantly recombinogenic. The highest concentrations were neither mutagenic nor recombinogenic, which could indicate cytotoxicity in the wing cells of D. melanogaster. In vitro, all BthTX-I concentrations (1 to 50 μg/mL) induced decrease in HUVEC cell viability, as well as in DU-145 cells at concentrations of 10, 25, and 50 μg/mL. The comet assay showed that in HUVEC and DU-145 cells, all BthTX-I concentrations promoted increase of DNA damage. Further studies should be performed to elucidate the mechanism of action of PLA2 BthTX-I and its possible use in therapeutic strategies against cancer.

Investigation of the Genotoxicity of Tatrazine and Anti-Genotoxicity of Some Plants Extracts in Drosophila Melanogaster Using the Smart Test

Food colorings are the food additives, which are used for improving the appearance of food and beverages. In the present study, genotoxic effects of Tatrtazine (E 102), is an synthetic food coloring, was investigated in vivo using the wing spot test, SMART (somatic mutation and recombination test), in Drosophila melanogaster Meigen. The food coloring at 25‰, 50‰ and 75‰ concentrations were added to food mediums of Drosophila during the larval stage and the numbers of mutant wing spots were evaluated using the SMART. Negative control medium was prepared with distilled water, while positive control medium was prepared with 1 mM EMS (ethyl methane sulfonate). According to results obtained from SMART, Tartrazine demonstrated significant results in trans-heterozygous flies (mwh/flr3) for inducing the mutant wing spots compared to control groups at 25 mg/ml, 50 mg/ml, and 75 mg/ml exposure concentrations. On the other hand, Tartrazine yielded significant results for inducing the mutant wing spots in balancer-heterozygous flies (mwh/TM3) at 25 mg/ml, 50 mg/ml, and 75 mg/ml exposure concentrations.The numbers of mutant wing spots were increased by the food coloring depending on the concentration (Chi-Square test; df=3, P<0.001). It was also determined that the numbers of mutant wing spots were significantly higher than the flies in the negative control medium and it suggests that Tartrazine has genotoxic effect. However, these numbers were less than the flies in the positive control medium; the data indicate that genotoxic effect of the food coloring was not as much as the EMS.On the other hand, the concentrations of Tartrazine were used in the mediums together the different plant extracts to determine whether they have anti-genotoxic effects against the food coloring. The following plant extracts were added to mediums at 100 mg/ml concentration with the food coloring was: Hypericum perforatum L., (St John’s wort), Silybum marianum (L.) Gaertn. (milk thistle), and Lavandula stoechas L. (topped lavender). When the mutant wing spots were compared to the control groups showed that the three plant extracts have no effect to reduce numbers of mutant wing spots in trans-heterozygous flies (mwh/flr3) and in balancer-heterozygous flies (mwh/TM3) and thus, have no anti-genotoxic effect against the food coloring used experimental treatments (Kruskal-Wallis test; df=2, P>0.05). Keywords: Genotoxicity, anti-genotoxicity, Tartrazine, Hypericum perforatum L., Silybum marianum (L.) Gaertn., Lavandula stoechas L., Drosophila melanogaster Meigen, SMART. DOI: 10.7176/JHMN/89-04 Publication date: May 31 st 2021

Genotoxicity and cytotoxicity evaluation of two thallium compounds using the Drosophila wing somatic mutation and recombination test

Thallium (Tl) is a heavy and toxic metal and a byproduct of several human activities, such as cement production, mining, and coal combustion. Thallium is found in fruits, vegetables, and animal fodder with high Tl contamination; therefore, it is an environmental pollution issue and a toxicological contamination problem for human beings and other organisms when exposed to it. The mutagenic potential of Tl and its compounds is controversial, and there are few in vivo studies on its effects. We conducted the animal bioassay Drosophila wing somatic mutation and recombination test (SMART) to test for genotoxicity and assessed the genotoxic effects of Tl acetate (TlCH3COO) and Tl sulfate (Tl2SO4) on Drosophila melanogaster. Third instar larvae from the SMART standard cross (ST) were fed Tl acetate [0.2, 2, 20, 200, 600 and 1200 μM] and Tl sulfate [0.2, 2, 20, 200, and 600 μM]. Hexavalent chromium [CrO3, 500 μM] served as the positive control, and Milli-Q water served as the negative control. Only the high Tl2SO4 [600 μM] concentration resulted in genotoxicity with 87.6% somatic recombination, and both salts disrupted cell division of wing imaginal disc cells, showing the expected cytotoxic effects. Genotoxic risks due to high metal levels by bioaccumulation of Tl+1 or its compounds require further evaluation with other in vivo and in vitro assays.

Adverse biological effects of ingested polystyrene microplastics using Drosophila melanogaster as a model in vivo organism

ABSTRACT The abundant presence and extensive use of polystyrene microplastics (PSMPs) has recently become a serious environmental concern, as impact of exposure to these substances on human health remains unknown. While in vitro studies yield data on adverse effect of PSMPs, in vivo approaches are more relevant for risk assessment. Drosophila melanogaster is one of the most genetically and experimentally accessible model organisms used in biology as an in vivo model. D. melanogaster was selected as a representative in vivo model organism to examine the genotoxic potential of PSMPs at 5 concentrations of three different sizes namely 4, 10, or 20 µm. In particular, the wing somatic mutation and recombination test (SMART), a scalable, time-efficient in vivo assay developed to study genotoxicity of various compounds in a rapid manner at low costs was used. The third–instar Drosophila larvae were exposed to PSMPs through food at 5 concentrations ranging from 0.01-10 mM. Viability (lethality), larval length, morphological deformations, locomotor activity (climbing behavior), and genotoxic effects were the end-points measured. Exposure to PSMPs at 4, 10, or 20 µm produced significant morphological defects, impaired climbing behavior, and genotoxicity as evidenced by the SMART test demonstrating induction of somatic recombination. Significant increases were observed in the frequency of total spots, suggesting that PSMPs might induce genotoxic activity predominantly via initiation of somatic DNA recombination in a concentration-dependent manner.

Somatic mutations and recombination test in Drosophila melanogaster used for investigating the genotoxicity of some food additives

In the present study, the effects of several food colorings, namely (Ponceau 4R (E 124), Tartrazine (E 102), and Pea green (E 102-E 133), were investigated in vivo using the wing spot test, SMART (somatic mutation and recombination test), in Drosophila melanogaster. Food colorings are the food additives, which are used for improving the appearance of food and beverages. In SMART, multiple wing hair (mwh), flare (flr3), and beaded serrate (BdS) marker genes on the third-largest chromosome of Drosophila are used. The genotoxic effects of the food colorings on the imaginal disc cells that will develop into the wing spot cells during the embryonic development of Drosophila heterozygous larvae and the genotypic changes caused by mutation or recombination in somatic cells also play a role in the formation of mutant spots in the wings. Classes by mutant clones are as follows: small single spots containing 1-2 mwh, large single spots containing ≥3 mwh or ≥4 flr3, and twin spots containing adjacent mwh and flr3 cells (GRAF et al., 1984). Negative control medium was prepared with distilled water, while positive control medium was prepared with 1 mM EMS (ethyl methane sulfonate). According to results obtained from SMART, Ponceau 4R, Tartrazine, and Pea green demonstrated significant results in trans-heterozygous flies (mwh/flr3) for inducing the mutant wing spots compared to control groups at 25 mg/ml, 50 mg/ml, and 75 mg/ml exposure concentrations. On the other hand, Ponceau 4R, Tartrazine, and Pea green yielded significant results for inducing the mutant wing spots in balancer-heterozygous flies (mwh/TM3) at 25 mg/ml, 50 mg/ml, and 75 mg/ml exposure concentrations. The numbers of mutant wing spots were increased by all three colorings depending on the concentration (X2= df=3, P<0.001). It was also determined that these numbers were significantly higher than the flies in the negative control medium and it suggests that these food colorings have genotoxic effects. However, the numbers of mutant wing spot were less than the flies in the positive control medium; this finding indicates that genotoxic effects of the food colorings were not as much as the EMS.

Investigation of The Genotoxicity of Ponceau 4R in Drosophila Melanogaster Using the Smart Test and Effects of Some Plants Extracts Against the DNA Damage

In this study, genotoxic effects of Ponceau 4R (E 124) was investigated in vivo using Drosophila melanogaster Meigen in the laboratory experiments. The food coloring in 25‰, 50‰ and 75‰ concentrations were added to food medium of Drosophila during the larval stage and the mutant wing spot numbers were evaluated using the wing spot test, namely SMART (somatic mutation and recombination test). The food colourings are the food additives which are used for increasing the appearance of food and beverages. In SMART, mwh (multiple wing hair), flr3 (flare) and BdS (beader serrate) marker genes on the third largest chromosome of Drosophila are used. Genotoxic effects of the Ponceau 4R in the imaginal disc cells that will develop into the wing spot cells during the embryonic development of Drosophila heterozygous larvae and the genotypic changes caused by mutation or recombination in somatic cells also play a role in the formation of mutant spots in the wings (GRAF et al., 1984). The food coloring which is daily used in our many types of food induced mutant wing spots depend on the concentration in trans-heterozygous flies (mwh/flr3) and in balancer-heterozygous flies (mwh/TM3) in the medium (Chi-Square test; df=3, P<0.001) and the spots were significantly higher than the flies were fed in the medium prepared with distilled water used as negative control showing that the Ponceau 4R has mutagenic effect. However, the mutant spots were less than the flies were fed in the medium prepared with 1 mM EMS (ethyl methane sulfonate) used as positive control showing that mutagenic effect of the colouring was not as much as the EMS. On the other hand, the concentrations of the colouring were used in the mediums together the different plant extracts to determinate whether they have antigenotoxic effects against the colouring. The following plant extracts were added to mediums in 10% concentration with the colouring were: Hypericum perforatum L., (St John’s wort), Silybum marianum (L.) Gaertn. (milk thistle), and Lavandula stoechas L. (topped lavender). The mutant wing spots were compared to control groups showed that the three plant extracts have no effect to reduce the mutant spots in trans-heterozygous flies (mwh/flr3) and in balancer-heterozygous flies and thus have no antigenotoxic effect against the food colouring used experimental treatments (Kruskal-Wallis test; df=2, P>0.05) Keywords: Challenge Genotoxicity, antigenotoxicity, Ponceau 4R, Hypericum perforatum L., Silybum marianum (L.) Gaertn., Lavandula stoechas L., Drosophila melanogaster Meigen, SMART. DOI: 10.7176/JHMN/87-07 Publication date: March 31 st 2021

An Evaluation of the Genotoxic Effects of Electromagnetic Radiation at 900 MHz, 1800 MHz, and 2100 MHz Frequencies with a SMART Assay in Drosophila melanogaster

ABSTRACT With the development of today’s technology, the electromagnetic radiation spread by mobile phones and base stations is also rapidly increasing, and this causes serious concerns about the environment and human health. The Drosophila model organism is widely used in genetic toxicology studies because its genome is highly similar to the genes identified in human diseases. In this study, the genotoxic effects of radiofrequency electromagnetic radiation were evaluated by the wing Somatic Mutation and Recombination Test (SMART) in Drosophila melanogaster at 900 MHz, 1800 MHz, and 2100 MHz. The SMART method is based on the observation of genetic changes occurring in the trichomes of the Drosophila wings appearing as mutant clones under the microscope. Throughout the study, total clone parameters were evaluated by exposing the Drosophila larvae to electromagnetic fields for two, four, and six hours per day for two days. As a result of the study, it was observed that the number of mutant clones was statistically increased according to the negative control group in all applications except for the six-hour application at 1800 MHz.