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Keywords = SCGE assay

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12 pages, 1068 KiB  
Article
Evaluation of Sperm DNA Fragmentation Using Two Different Methods: TUNEL via Fluorescence Microscopy, and Flow Cytometry
by Katerina Chatzimeletiou, Alexandra Fleva, Theodoros-Thomas Nikolopoulos, Maria Markopoulou, Glykeria Zervakakou, Kyriakos Papanikolaou, George Anifandis, Anastasia Gianakou and Grigoris Grimbizis
Medicina 2023, 59(7), 1313; https://doi.org/10.3390/medicina59071313 - 15 Jul 2023
Cited by 8 | Viewed by 6085
Abstract
Background and Objectives: Sperm DNA fragmentation refers to any break in one or both of the strands of DNA in the head of a sperm. The most widely used methodologies for assessing sperm DNA fragmentation are the sperm chromatin structure assay (SCSA), [...] Read more.
Background and Objectives: Sperm DNA fragmentation refers to any break in one or both of the strands of DNA in the head of a sperm. The most widely used methodologies for assessing sperm DNA fragmentation are the sperm chromatin structure assay (SCSA), the sperm chromatin dispersion assay (SCD), the single-cell gel electrophoresis assay (SCGE–comet), and the terminal-deoxynucleotidyl-transferase (TdT)-mediated dUTP nick end labelling (TUNEL) assay. The aim of this study was to compare the efficiency and sensitivity of the analysis of sperm DNA fragmentation using TUNEL via fluorescence microscopy, and flow cytometry. Materials and Methods: Semen samples were collected and analyzed for standard characteristics using light microscopy, and for sperm DNA fragmentation using both TUNEL via fluorescence microscopy, and flow cytometry. Results: There were no significant differences in the values of the sperm DNA fragmentation index (DFI) obtained when the analysis was performed using TUNEL or flow cytometry (p = 0.543). Spearman’s correlation analysis revealed a significant negative correlation between sperm motility (%) and sperm DNA fragmentation (p < 0.01), as well as between sperm concentration and sperm DNA fragmentation (p < 0.05). The Mann–Whitney U test showed no significant difference in the DFI among couples with repeated implantation failure (RIF) and miscarriages (p = 0.352). Conclusions: Both methods (TUNEL via fluorescence microscopy, and flow cytometry) have a high efficiency and sensitivity in accurately detecting sperm DNA fragmentation, and can be effectively used to assess male fertility. Full article
(This article belongs to the Section Obstetrics and Gynecology)
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15 pages, 687 KiB  
Article
Effect of Selected Micro- and Macroelements and Vitamins on the Genome Stability of Bovine Embryo Transfer Recipients following In Vitro Fertilization
by Ewa Wójcik, Katarzyna Kępka and Mateusz Skup
Animals 2023, 13(6), 1056; https://doi.org/10.3390/ani13061056 - 14 Mar 2023
Cited by 4 | Viewed by 2608
Abstract
Genome instability can lead to a wide variety of diseases. Many endogenous and exogenous factors influence the level of damage to genetic material. Genome integrity depends on factors such as the fidelity of DNA replication, normal DNA organization in the chromosomes, and repair [...] Read more.
Genome instability can lead to a wide variety of diseases. Many endogenous and exogenous factors influence the level of damage to genetic material. Genome integrity depends on factors such as the fidelity of DNA replication, normal DNA organization in the chromosomes, and repair mechanisms. Genome stability influences fertility, embryonic development, and the maintenance of pregnancy. In the case of in vitro fertilization, it can be an important factor determining the success of the procedure. The aim of the study was to assess the stability of the genomes of recipient cows following in vitro fertilization using cytogenetic tests and to analyze the effects of selected vitamins and micro- and macroelements on genome integrity. Genome stability was analyzed using the sister chromatid exchange, fragile site, and comet assays. The material for analysis was peripheral blood from 20 Holstein-Friesian heifers that were embryo transfer recipients. The effect of selected micro- and macroelements and vitamins on the genome stability of the cows was analyzed. Folic acid was shown to significantly influence the level of damage identified using the SCE, FS, and SCGE assays, while iron affected SCE and SCGE results, and zinc affected FS. Full article
(This article belongs to the Collection Advances in Cattle Breeding, Genetics and Genomics)
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42 pages, 1258 KiB  
Review
An Overview of Comet Assay Application for Detecting DNA Damage in Aquatic Animals
by Nan Jiang, Saima Naz, Yulin Ma, Qudrat Ullah, Muhammad Zahoor Khan, Jiaqi Wang, Xuan Lu, Dun-Zhu Luosang, Sadia Tabassum, Ahmad Manan Mustafa Chatha and Wang-Dui Basang
Agriculture 2023, 13(3), 623; https://doi.org/10.3390/agriculture13030623 - 5 Mar 2023
Cited by 36 | Viewed by 14449
Abstract
This review discusses several research studies that employed comet assay to evaluate the environmental impact of genotoxins in aquatic environments. It focuses on in vivo and in situ studies of aquatic animals. New chemicals are being added each year to the existing burden [...] Read more.
This review discusses several research studies that employed comet assay to evaluate the environmental impact of genotoxins in aquatic environments. It focuses on in vivo and in situ studies of aquatic animals. New chemicals are being added each year to the existing burden of toxic substances in the environment. Excessive agricultural and industrial activities adversely affect biodiversity, threatening the survival of species in a particular habitat, as well as posing disease risks to humans. Some of the chemicals, e.g., pesticides and heavy metals, may be genotoxic to the sentinel species and/or to non-target species, causing deleterious effects in somatic or germ cells. Comet assay is a quick, sensitive, and low-cost technique for detecting DNA strand breakage. However, the comet assay has much more to offer than being an assay for testing DNA strand breaks in animal organs. The use of repair enzymes increases the range of DNA lesions that can be detected with the assay. Comparing data from studies that employed different approaches, such as empirical scoring or comet tail lengths, comet assay is one of the challenging techniques to be utilized in environmental studies. The relative amount of DNA in the comet tail indicates DNA break intensity. The assay has been modified to detect various base alterations by including the digestion of nucleoids with a lesion-specific endonuclease. The determination of DNA damage in these indicator species using the comet test would thus offer information on the genotoxic potential of their habitat at an early stage. This would enable intervention techniques to prevent or mitigate adverse health impacts in sentinel animals and humans. Full article
(This article belongs to the Special Issue Sustainable Aquaculture: Current Perspectives and Future Challenges)
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39 pages, 16985 KiB  
Article
Boron Compounds Exhibit Protective Effects against Aluminum-Induced Neurotoxicity and Genotoxicity: In Vitro and In Vivo Study
by Hasan Turkez, Serkan Yıldırım, Elvan Sahin, Mehmet Enes Arslan, Bugrahan Emsen, Ozlem Ozdemir Tozlu, Gonca Alak, Arzu Ucar, Abdulgani Tatar, Ahmet Hacimuftuoglu, Mevlut Sait Keles, Fatime Geyikoglu, Muhammed Atamanalp, Fatih Saruhan and Adil Mardinoglu
Toxics 2022, 10(8), 428; https://doi.org/10.3390/toxics10080428 - 28 Jul 2022
Cited by 29 | Viewed by 4868
Abstract
Genetic, neuropathological and biochemical investigations have revealed meaningful relationships between aluminum (Al) exposure and neurotoxic and hematotoxic damage. Hence, intensive efforts are being made to minimize the harmful effects of Al. Moreover, boron compounds are used in a broad mix of industries, from [...] Read more.
Genetic, neuropathological and biochemical investigations have revealed meaningful relationships between aluminum (Al) exposure and neurotoxic and hematotoxic damage. Hence, intensive efforts are being made to minimize the harmful effects of Al. Moreover, boron compounds are used in a broad mix of industries, from cosmetics and pharmaceuticals to agriculture. They affect critical biological functions in cellular events and enzymatic reactions, as well as endocrinal and mineral metabolisms. There are limited dose-related data about boric acid (BA) and other boron compounds, including colemanite (Col), ulexite (UX) and borax (BX), which have commercial prominence. In this study, we evaluate boron compounds’ genetic, cytological, biochemical and pathological effects against aluminum chloride (AlCl3)-induced hematotoxicity and neurotoxicity on different cell and animal model systems. First, we perform genotoxicity studies on in vivo rat bone marrow cells and peripheric human blood cultures. To analyze DNA and chromosome damage, we use single cell gel electrophoresis (SCGE or comet assay) and micronucleus (MN) and chromosome aberration (CA) assays. The nuclear division index (NDI) is used to monitor cytostasis. Second, we examine the biochemical parameters (superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), malondialdehyde (MDA), total antioxidant capacity (TAC) and total oxidative status (TOS)) to determine oxidative changes in blood and brain. Next, we assess the histopathological alterations by using light and electron microscopes. Our results show that Al increases oxidative stress and genetic damage in blood and brain in vivo and in vitro studies. Al also led to severe histopathological and ultrastructural alterations in the brain. However, the boron compounds alone did not cause adverse changes based on the above-studied parameters. Moreover, these compounds exhibit different levels of beneficial effects by removing the harmful impact of Al. The antioxidant, antigenotoxic and cytoprotective effects of boron compounds against Al-induced damage indicate that boron may have a high potential for use in medical purposes in humans. In conclusion, our analysis suggests that boron compounds (especially BA, BX and UX) can be administered to subjects to prevent neurodegenerative and hematological disorders at determined doses. Full article
(This article belongs to the Section Ecotoxicology)
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22 pages, 393 KiB  
Systematic Review
Genotoxicity of Mercury and Its Derivatives Demonstrated In Vitro and In Vivo in Human Populations Studies. Systematic Review
by Juana Sánchez-Alarcón, Mirta Milić, Lilia Patricia Bustamante-Montes, Keila Isaac-Olivé, Rafael Valencia-Quintana and Ninfa Ramírez-Durán
Toxics 2021, 9(12), 326; https://doi.org/10.3390/toxics9120326 - 1 Dec 2021
Cited by 16 | Viewed by 3919
Abstract
Beside partial coverage in three reviews so far (1994, 2009, 2019), there is no review on genotoxic studies dealing with mercury (Hg) and human exposure using the most usual genotoxic assays: sister chromatid exchanges (SCE), chromosomal aberrations (CA), cytochalasin B blocked micronucleus assay [...] Read more.
Beside partial coverage in three reviews so far (1994, 2009, 2019), there is no review on genotoxic studies dealing with mercury (Hg) and human exposure using the most usual genotoxic assays: sister chromatid exchanges (SCE), chromosomal aberrations (CA), cytochalasin B blocked micronucleus assay (CBMN), and single-cell gel electrophoresis (SCGE or alkaline comet assay). Fifty years from the first Hg genotoxicity study and with the Minamata Convention in force, the genotoxic potential of Hg and its derivatives is still controversial. Considering these antecedents, we present this first systematic literature overview of genotoxic studies dealing with Hg and human exposure that used the standard genotoxic assays. To date, there is not sufficient evidence for Hg human carcinogen classification, so the new data collections can be of great help. A review was made of the studies available (those published before the end of October 2021 on PubMed or Web of Science in English or Spanish language) in the scientific literature dealing with genotoxic assays and human sample exposure ex vivo, in vivo, and in vitro. Results from a total of 66 articles selected are presented. Organic (o)Hg compounds were more toxic than inorganic and/or elemental ones, without ruling out that all represent a risk. The most studied inorganic (i)Hg compounds in populations exposed accidentally, occupationally, or iatrogenically, and/or in human cells, were Hg chloride and Hg nitrate and of the organic compounds, were methylmercury, thimerosal, methylmercury chloride, phenylmercuric acetate, and methylmercury hydroxide. Full article
17 pages, 5000 KiB  
Review
Genotoxicity Assessment of Metal-Based Nanocomposites Applied in Drug Delivery
by Sara Cardoso, Classius F. da Silva, Patrícia Severino, Amélia M. Silva, Selma B. Souto, Aleksandra Zielińska, Jacek Karczewski and Eliana B. Souto
Materials 2021, 14(21), 6551; https://doi.org/10.3390/ma14216551 - 1 Nov 2021
Cited by 6 | Viewed by 3189
Abstract
Nanocomposites as drug delivery systems (e.g., metal nanoparticles) are being exploited for several applications in the biomedical field, from therapeutics to diagnostics. Green nanocomposites stand for nanoparticles of biocompatible, biodegradable and non-toxic profiles. When using metal nanoparticles for drug delivery, the question of [...] Read more.
Nanocomposites as drug delivery systems (e.g., metal nanoparticles) are being exploited for several applications in the biomedical field, from therapeutics to diagnostics. Green nanocomposites stand for nanoparticles of biocompatible, biodegradable and non-toxic profiles. When using metal nanoparticles for drug delivery, the question of how hazardous these “virus-sized particles” can be is posed, due to their nanometer size range with enhanced reactivity compared to their respective bulk counterparts. These structures exhibit a high risk of being internalized by cells and interacting with the genetic material, with the possibility of inducing DNA damage. The Comet Assay, or Single-Cell Gel Electrophoresis (SCGE), stands out for its capacity to detect DNA strand breaks in eukaryotic cells. It has huge potential in the genotoxicity assessment of nanoparticles and respective cells’ interactions. In this review, the Comet assay is described, discussing several examples of its application in the genotoxicity evaluation of nanoparticles commonly administered in a set of routes (oral, skin, inhaled, ocular and parenteral administration). In the nanoparticles boom era, where guidelines for their evaluation are still very limited, it is urgent to ensure their safety, alongside their quality and efficacy. Comet assay or SCGE can be considered an essential tool and a reliable source to achieve a better nanotoxicology assessment of metal nanoparticles used in drug delivery. Full article
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12 pages, 3220 KiB  
Article
In Vitro Probiotic Properties and DNA Protection Activity of Yeast and Lactic Acid Bacteria Isolated from A Honey-Based Kefir Beverage
by Bruna de Oliveira Coelho, Fernanda Fiorda-Mello, Gilberto V. de Melo Pereira, Vanete Thomaz-Soccol, Sudip K. Rakshit, Júlio Cesar de Carvalho and Carlos Ricardo Soccol
Foods 2019, 8(10), 485; https://doi.org/10.3390/foods8100485 - 12 Oct 2019
Cited by 43 | Viewed by 6082
Abstract
The probiotic characteristics of three acid-tolerant microbial strains, viz., Lactobacillus satsumensis LPBF1, Leuconostoc mesenteroides LPBF2 and Saccharomyes cerevisiae LPBF3, isolated from a honey-based kefir functional beverage, were studied following the requirements established by the Food and Agriculture Organization of the United Nation/World Health [...] Read more.
The probiotic characteristics of three acid-tolerant microbial strains, viz., Lactobacillus satsumensis LPBF1, Leuconostoc mesenteroides LPBF2 and Saccharomyes cerevisiae LPBF3, isolated from a honey-based kefir functional beverage, were studied following the requirements established by the Food and Agriculture Organization of the United Nation/World Health Organization (FAO/WHO), including host-associated stress resistance, epithelium adhesion ability, and antimicrobial activity. The three microbial strains tolerated different pH values (2.0, 3.0, 4.0 and 7.0) and bile salt concentrations (0.3% and 0.6%), and survive in the presence of simulated gastric juice, which are conditions imposed by the gastrointestinal tract. In addition, they showed high percentages of hydrophobicity, auto aggregation and anti-pathogenic against Escherichia coli and Staphylococcus aureus, with no hemolytic activity. The protective capacity of human DNA through microbial treatment was investigated by single-cell gel electrophoresis (SCGE) comet assay. The three selected strains showed DNA protection effect against damage caused by hydroxyl radical (H2O2). However, when the S. cerevisiae treatment was applied, the most effective DNA protection index was observed, which can be associated to its high production of extracellular antioxidants as reveled by the 2,2-diphenyl-1-picryl-hydrazylhydrate (DPPH) method. These results indicated that the three selected microbial strains could be useful for preventing oxidative DNA damage and cellular oxidation in food products. As well-adapted microbial cells, the selected strains can be used for production of non-dairy functional beverages, especially for vegans and/or vegetarians and lactose intolerants. Full article
(This article belongs to the Special Issue Milk Alternatives and Non-Dairy Fermented Products)
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20 pages, 1272 KiB  
Article
Vitamin E Modifies High-Fat Diet-Induced Increase of DNA Strand Breaks, and Changes in Expression and DNA Methylation of Dnmt1 and MLH1 in C57BL/6J Male Mice
by Marlene Remely, Franziska Ferk, Sonja Sterneder, Tahereh Setayesh, Tatjana Kepcija, Sylvia Roth, Rahil Noorizadeh, Martina Greunz, Irene Rebhan, Karl-Heinz Wagner, Siegfried Knasmüller and Alexander Haslberger
Nutrients 2017, 9(6), 607; https://doi.org/10.3390/nu9060607 - 14 Jun 2017
Cited by 49 | Viewed by 6858
Abstract
Obesity is associated with low-grade inflammation, increased ROS production and DNA damage. Supplementation with antioxidants might ameliorate DNA damage and support epigenetic regulation of DNA repair. C57BL/6J male mice were fed a high-fat (HFD) or a control diet (CD) with and without vitamin [...] Read more.
Obesity is associated with low-grade inflammation, increased ROS production and DNA damage. Supplementation with antioxidants might ameliorate DNA damage and support epigenetic regulation of DNA repair. C57BL/6J male mice were fed a high-fat (HFD) or a control diet (CD) with and without vitamin E supplementation (4.5 mg/kg body weight (b.w.)) for four months. DNA damage, DNA promoter methylation and gene expression of Dnmt1 and a DNA repair gene (MLH1) were assayed in liver and colon. The HFD resulted in organ specific changes in DNA damage, the epigenetically important Dnmt1 gene, and the DNA repair gene MLH1. Vitamin E reduced DNA damage and showed organ-specific effects on MLH1 and Dnmt1 gene expression and methylation. These results suggest that interventions with antioxidants and epigenetic active food ingredients should be developed as an effective prevention for obesity—and oxidative stress—induced health risks. Full article
(This article belongs to the Special Issue Immunology: Nutrition, Exercise and Adiposity Relationships)
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8 pages, 167 KiB  
Article
Genotoxicity of Air Borne Particulates Assessed by Comet and the Salmonella Mutagenicity Test in Jeddah, Saudi Arabia
by Sufian M. ElAssouli, Mohamed H. AlQahtani and Waleed Milaat
Int. J. Environ. Res. Public Health 2007, 4(3), 216-223; https://doi.org/10.3390/ijerph2007030004 - 30 Sep 2007
Cited by 35 | Viewed by 10888
Abstract
Fine airborne respirable particulates less than 10 micrometer (PM10) are considered one of the top environmental public health concerns, since they contain polycyclic aromatic hydrocarbons (PAHs) which are among the major carcinogenic compounds found in urban air. The objective of this study is [...] Read more.
Fine airborne respirable particulates less than 10 micrometer (PM10) are considered one of the top environmental public health concerns, since they contain polycyclic aromatic hydrocarbons (PAHs) which are among the major carcinogenic compounds found in urban air. The objective of this study is to assess the genotoxicity of the ambient PM10 collected at 11 urban sites in Jeddah, Saudi Arabia. The PM10 extractable organic matter (EOM) was examined for its genotoxicity by the single cell gel electrophoresis (SCGE) comet assay and the Salmonella mutagenicity (Ames) test .Gas chromatography-mass spectrometry was used to quantify 16 PAH compounds in four sites. Samples from oil refinery and heavy diesel vehicles traffic sites showed significant DNA damage causing comet in 20-44% of the cells with tail moments ranging from 0.5-2.0 compared to samples from petrol driven cars and residential area, with comet in less than 2% of the cells and tail moments of < 0.02.In the Ames test, polluted sites showed indirect mutagenic response and caused 20-56 rev/ m3, mean while residential and reference sites caused 2-15 rev /m3. The genotoxicity of the EOM in both tests directly correlated with the amount of organic particulate and the PAHs concentrations in the air samples. The PAHs concentrations ranged between 0.83 ng/m3 in industrial and heavy diesel vehicles traffic sites to 0.18 ng /m3 in the residential area. Benzo(ghi)pyrene was the major PAH components and at one site it represented 65.4 % of the total PAHs. Samples of the oil refinery site were more genotoxic in the SCGE assay than samples from the heavy diesel vehicles traffic site, despite the fact that both sites contain almost similar amount of PAHs. The opposite was true for the mutagenicity in the Ames test. This could be due to the nature of the EOM in both sites. These findings confirm the genotoxic potency of the PM10 organic extracts to which urban populations are exposed. Full article
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