Comet Tail Length Research Articles

Abstract Mo036: DNA Damage and Repair in Patients Undergoing Myocardial Perfusion Single-Photon Emission Computed Tomography

Background: As medical imaging has continuously grown, patient exposure to ionizing radiation and the risks associated with it are important issues. Myocardial perfusion single-photon emission computed tomography (MP-SPECT) is one of the most frequently performed noninvasive cardiac imaging tests, generating concern about the potentially excessive radiation exposure for patients, including the cumulative doses of lifetime radiation. even though the individual levels of radiation exposure are considered low. Aim: To evaluate DNA damage and DNA repair markers after MP-SPECT. Methods: Thirty-two patients undergoing Tc-99m sestamibi MP-SPECT (mean injected Tc-99m sestamibi dose: 15.1 mCi) were studied. Peripheral blood was collected before radiotracer injection at rest and 60-90 minutes after injection. The comet assay (single-cell gel electrophoresis) was performed with blood lymphocytes to detect DNA strand breaks, which appear as DNA fragmentation (the comet tail; Figure1). Images of fluorescence microscopy were analyzed using the Comet Assay Software Project Lab (CASPLab 1.2.3 beta 2). Three parameters were evaluated: the percentage of DNA in the comet tail, tail length, and tail moment (the measure of migratory DNA and number of fragments). Quantitative PCR (qPCR) was performed to evaluate the expression of five genes related to signaling pathways in response to DNA damage and repair (ATM, ATR, BRCA1, CDKN1A, XPC). Data are expressed as medians, and Wilcoxon or Mann-Whitney tests were employed for statistical analysis. A p-value <0.05 was considered statistically significant. Results: Figure 2 shows pre- and post- radiation exposure comet assay parameters. After radiotracer injection, there was a 10.7% increase of the DNA percentage in the tail (pre-exposure median 2.185 vs post-exposure median 2.418, p<0.0065). Tail length increased 6.7% (15.0 pre-exposure vs 16.0 post-exposure, p<0.0034). Tail moment increased 27.5% (0.3177 pre-exposure vs 0.4050 post-exposure, p<0.0001). qPCR did not reveal increased expression of any gene after radiation exposure. Conclusion: An increase in DNA damage was detected after a single radiotracer injection for MP-SPECT, yet unaccompanied by activation of repair genes. This may suggest that DNA damage is not large enough to trigger intense repair responses and might not result in biologically significant DNA damage.

DNA damage and repair in patients undergoing myocardial perfusion single-photon emission computed tomography

As patient exposure to ionizing radiation from medical imaging and its risks are continuing issues, this study aimed to evaluate DNA damage and repair markers after myocardial perfusion single-photon emission computed tomography (MPS). Thirty-two patients undergoing Tc-99m sestamibi MPS were studied. Peripheral blood was collected before radiotracer injection at rest and 60–90 min after injection. The comet assay (single-cell gel electrophoresis) was performed with peripheral blood cells to detect DNA strand breaks. Three descriptors were evaluated: the percentage of DNA in the comet tail, tail length, and tail moment (the product of DNA tail percentage and tail length). Quantitative PCR (qPCR) was performed to evaluate the expression of five genes related to signaling pathways in response to DNA damage and repair (ATM, ATR, BRCA1, CDKN1A, and XPC). Mann–Whitney’s test was employed for statistical analysis; p < 0.05 was considered significant. Mean Tc-99m sestamibi dose was 15.1 mCi. After radiotracer injection, comparing post-exposure to pre-exposure samples of each of the 32 patients, no statistically significant differences of the DNA percentage in the tail, tail length or tail moment were found. qPCR revealed increased expression of BRCA1 and XPC, without any significant difference regarding the other genes. No significant increase in DNA strand breaks was detected after a single radiotracer injection for MPS. There was activation of only two repair genes, which may indicate that, in the current patient sample, the effects of ionizing radiation on the DNA were not large enough to trigger intense repair responses, suggesting the absence of significant DNA damage.

IJCM_265A: Cellular Consequences of Fruit Juice Metallurgy: A Closer Look at Heavy Metal Influence

Background: The leaching of heavy metals from packaging materials into packed juices can have several adverse effects on the quality and safety of the products and potentially pose health risks to consumers. Objective: The research aimed to investigate the cytotoxic effects of heavy metals specifically vanadium, cadmium, arsenic, and mercury, which are commonly found in packaging materials of marketable fruit juices which are leached into juices itself (orange and mixed fruit juices stored in cans, leak- proof bags, and tetraPaks). Methodology: The study utilized cell lines, namely HepG2 (human liver cancer) and HeK293 (human kidney), to assess the impact of these heavy metals on cellular viability, DNA damage, mitochondrial function, and cell cycle progression. Flow cytometry was used to analyse cell cycle alterations. Results: The MTT assay showcased dose-dependent decrease in cell viability with increasing concentrations of heavy metals. Annexin V Assay determined V, Hg and As had necrotic death pattern while Cd had apoptotic death pattern for both the cell lines in presence of heavy metal digests. Cells treated with heavy metals, and the resulting DNA damage observed to be high after quantified using comet tail length, olive tail movement, and tail moment measurements. Mitochondrial function assays in the presence of heavy metals revealed that increased mitochondrial membrane potential, mitochondrial mass, and reactive oxygen species production. ROS generation assessed to understand the impact of heavy metals on cellular health and oxidative stress. Conclusion: This study shed light on the potential cytotoxic effects of heavy metals found in fruit juices. These findings contribute to our understanding of the potential risks associated with heavy metal exposure from consumer products and emphasize the importance of regulatory measures to ensure food safety and human health.

Comparison of DNA damage in granulosa cells of women undergoing controlled ovarian stimulation in in vitro fertilization protocols with the recombinant human follicle-stimulating hormones Corneumon®, Gonal-F®, Pergoveris® and Puregon®: a randomized trial.

To compare the DNA damage in granulosa cells (GCs) of women undergoing ovarian-stimulated cycles with four widely used recombinant human follicle-stimulating hormones (rhFSH) in in vitro fertilization (IVF) protocols (Corneumon®, Gonal-F®, Pergoveris® and Puregon®). A randomized trial was carried out at a Mexican hospital. GCs were isolated from 18 women with infertility undergoing assisted reproductive techniques (ART). Four controlled ovarian stimulation (COS) protocols including Corneumon®, Gonal-F®, Pergoveris® or Puregon® were used. GCs DNA damage was assessed by the Comet assay. Two parameters were measured: comet tail length (CTL), and Olive tail moment (OTM, the percentage of DNA in the tail multiplied by the distance between the center of the tail and head). Use of the different hrFSH in COS caused variable and statistically significant levels of DNA damage in GCs of infertile women. CTL was similar in the Corneumon® and Pergoveris® groups (mean values of 48.73 and 55.18, respectively) and Corneumon® CTL was significantly lower compared to the Gonal-F® and Puregon® groups (mean values of 61.98 and 91.17, respectively). Mean OTM values were significantly lower in Corneumon® and Pergoveris® groups, compared to Gonal-F® and Puregon® groups (25.59, 27.35, 34.76, and 47.27, respectively). Use of Corneumon® and Pergoveris® in COS caused statistically significantly lower levels of DNA damage in GCs of infertile women undergoing ART, which could potentially correlate with better reproductive outcomes.

Detection of DNA Damage in Fish using Comet Assay

Heavy metals have an enduring presence, risky characteristics, and the propensity to accumulate in the environment. This is why heavy metal toxics are widely acknowledged as harmful environmental pollutants. Heavy metals damage both aquatic and terrestrial ecosystems, posing a major risk to the environment and human health. Four freshwater fish species namely Labeo rohita, Catla catla, Hypophthalmichthys molitrix, and Ctenopharyngodon idella were the focus of this investigation. This study investigated the potential genotoxic effects of lead (Pb), copper (Cu), and cadmium (Cd) on the above fish species through the application of comet assay test. The fish were exposed to these metals at four distinct concentrations (19%, 24%, 31%, and 50% of the LC50) over the course of 40 days. All four fish species were exposed to metals to varying degrees, according to the genetic damage index, cumulative tail length of comets, and the proportion of damaged cells. In contrast to Catla catla, Hypophthalmichthys molitrix had the highest prevalence of DNA damage. The current study suggests that the presence of these particular metals in Pakistan's aquatic ecosystems may have an adverse effect on the DNA of the country's fish species. Metals cause damage to DNA in fibroblast cells through distinct mechanisms when present in water, air, and soil. Comet assay test has a remarkable sensitivity that helps to identify extremely low amounts of DNA damage. Out of the four fish species, Ctenopharyngodon idella showed higher levels of damaged cells, a higher genetic damage index, and a cumulative comet tail length as compared to others. All four fish species experienced a significant increase in DNA damage, genetic damage index, and comet tail length at 50% concentration of metals LC50.

The optimum concentration of soya-lecithin for replacing egg yolk in INRA-82-modified semen extenders improved the cryopreserved semen quality and DNA integrity of Arabian stallions

The cryopreservation of stallion spermatozoa varies greatly between animals. Different equine semen extender supplements were invented to maximize the post-thaw semen quality. Soy lecithin is one of those additives that contain plant origin phospholipids used to replace egg yolk to minimize disease transmission. This study aimed to determine which concentrations of soy lecithin can replace egg-yolk in modified INRA-82 semen extenders for the cryopreservation of horse semen. Semen was collected from six Arabian stallions and extended with modified INRA-82 semen extender supplemented with 0.0, 1.25, 2.50, and 5.0% soya lecithin. Frozen-thawed sperm motility was measured at 0, 1, 2, and 3 h. Viability index, sperm plasma membrane integrity (SPMI), and acrosome integrity in addition to the comet assay non-fragmented DNA, head DNA, tail DNA, tail length, and tail moment were statistically analyzed. The addition of 2.5% soya lecithin had the highest post-thaw sperm motility at 0, 1, 2 h (P &lt; 0.001) and 3 h (P &lt; 0.0001), the optimum viability index (P &lt; 0.0001), SPMI (P &lt; 0.05), acrosome integrity (P &lt; 0.0001), non-fragmented DNA (P &lt; 0.01), DNA in the comet head (P &lt; 0.05), minimum DNA in the comet tail (P &lt; 0.05), shortest comet tail length (P &lt; 0.0001), and lowest comet tail moment (P &lt; 0.01). Soy lecithin at concentrations other than 2.5% deteriorated all post-thaw semen parameters. Soy lecithin (2.5%) can replace egg yolk in the modified INRA-82 semen extender for the cryopreservation of good-quality stallion semen.

Waste anesthetic gases have a significant association with deoxyribonucleic acid (DNA) damage: A systematic review and meta-analysis of 2,732 participants

IntroductionOperating room workers are at risk of experiencing adverse effects due to occupational exposure to waste anesthetic gases (WAGs). One of the consequences of long-term WAGs exposure is the probability of developing deoxyribonucleic acid (DNA) damage. This systematic review investigated the link between WAGs and DNA damage in operating room workers. MethodsPubMed, Science Direct, ProQuest, Scopus, and EbscoHost, as well as hand-searching, were used to find literature on the relationship between WAGs and DNA damage. Three independent reviewers independently assessed the study's quality. Meta-analysis was conducted for several DNA damage indicators, such as comet assay (DNA damage score, tail's length, tail's DNA percentage), micronuclei formation, and total chromosomal aberration. ResultsThis systematic review included 29 eligible studies (2732 participants). The majority of the studies used a cross-sectional design. From our meta-analysis, which compared the extent of DNA damage in operating room workers to the unexposed group, operating room workers exposed to WAGs had a significantly higher DNA damage indicator, including DNA damage score, comet tail's length, comet tail's DNA percentage, micronuclei formation, and total chromosomal aberration (p < 0.05) than non-exposed group. ConclusionWaste anesthetic gases have been found to significantly impact DNA damage indicators in operating room personnel, including comet assay, micronuclei development, and chromosomal aberration. To reduce the impact of exposure, hospital and operating room personnel should take preventive measures, such as by adapting scavenger method.

Genotoxicity and genomic instability in oral epithelial cells of agricultural workers exposed to pesticides using micronucleus and comet assay in Nineveh, Iraq

In agriculture, pesticides are used to preserve plants, but they might be dangerous for farmers and the environment. The present study aimed to use the comet assay and the micronucleus (MN) test to assess the genotoxic effects on lymphocytes and buccal exfoliation in pesticide-exposed male agricultural workers. The samples were collected from 102 workers having exposure to pesticides (Roundup SL, Weed waster, and paraquat 20% SL) and 100 control individuals (without pesticide exposure) from different Mosul, Iraq, neighbourhoods. With the help of the comet assay and the MN test, exfoliated buccal cells from the individuals were analyzed for DNA damage. Each individual's lymphocytes and epithelial baseline cells had their comet tail length assessed, along with any other nuclear abnormalities such as nuclear buds, karyolysis, karyorrhexis, and binucleate cells. The results showed that the frequency of MN considerably rose in the exposed group, and that group also revealed nuclear anomalies linked to cytotoxic or genotoxic effects. There were significant disparities in the amount of DNA damage between recently exposed employees and controls and recently exposed and followed-up cases. In comparison to controls, there was a considerable increase in the and frequency of cells that migrated in exposed workers. However, it was shown that confounding factors, such as age and the varying length of pesticide exposure, substantially impacted DNA damage. Educational programs for agricultural workers are critical to limit the use of chemicals in agriculture, given the evidence of a genetic risk associated with exposure brought on by the extensive use of pesticides.

Biogenic Synthesis of Cu-Mn Bimetallic Nanoparticles Using Pumpkin Seeds Extract and Their Characterization and Anticancer Efficacy

Cancer is a chronic, heterogeneous illness that progresses through a spectrum of devastating clinical manifestations and remains the 2nd leading contributor to global mortality. Current cancer therapeutics display various drawbacks that result in inefficient management. The present study is intended to evaluate the anticancer potential of Cu-Mn bimetallic NPs (CMBNPs) synthesized from pumpkin seed extract against colon adenocarcinoma cancer cell line (HT-29). The CMBNPs were biosynthesized by continuously stirring an aqueous solution of pumpkin seed extract with CuSO4 and manganese (II) acetate tetrahydrate until a dark green solution was obtained. The characteristic features of biogenic CMBNPs were assessed by UV-visible spectrophotometry (UV-vis), X-ray powder diffraction (XRD), energy-dispersive X-ray (EDX), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). A battery of biological assays, viz. neutral red uptake (NRU) assay, in vitro scratch assay, and comet assay, were performed for anticancer efficacy evaluation. The formation of spherical monodispersed bimetallic nanoparticles with an average size of 50 nm was recorded using TEM. We observed dose-dependent cytotoxicity of CMBNPs in the HT-29 cell line with an IC50 dose of 115.2 µg/mL. On the other hand, CMBNPs did not show significant cytotoxicity against normal cell lines (Vero cells). Furthermore, the treatment of CMBNPs inhibited the migration of cancer cells and caused DNA damage with a significant increase in comet tail length. The results showed substantial anticancer efficacy of CMBNPs against the studied cancer cell line. However, it is advocated that the current work be expanded to different in vitro cancer models so that an in vivo validation could be carried out in the most appropriate cancer model.

An Overview of Comet Assay Application for Detecting DNA Damage in Aquatic Animals

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.

Elucidation of the cytogenotoxic potential of vigabatrin and its in silico computer-assisted DNA interaction

Vigabatrin (VGB) is a gammaaminobutyric acid-ergic (GABA-ergic) antiepileptic drug (AED) and is one of 2 approved drugs available to treat infantile spasms (IS). The aim of this study is to elucidate conflicting data on the toxic effects of VGB and to obtain detailed information about its possible cytogenotoxic effects in human lymphocytes. For this purpose, in vitro Chromosomal Aberration (CA), Sister Chromatid Exchange (SCE), Micronucleus (MN) tests, and Comet Assay were performed to determine possible genotoxic and cytotoxic effects of VGB. In addition, the binding energy level of VGB to DNA was determined in silico by molecular docking. The highest concentration (80 μg/ml) of VGB increased the SCE, CA, MN and micronucleated binuclear cell (BNMN) frequency significantly compared to the control after 24 and 48 hours of treatment. In the tail density and tail length parameters, the dose-dependent increase was found to be statistically significant compared to the control. At the 40 and 80 μg/ml concentrations of VGB for 48 hours caused a statistically significant increase in both CA/Cell and AC percentages, while MI and NDI decreased only significantly at the highest concentration (80 µg/ml) causing. In the Comet Assay head density, tail density and tail length parameters, the dose-dependent increase was found to be statistically significant compared to the control. Also, the in silico molecular docking analysis showed that VGB interacts with B-DNA close to the threshold binding energy. The lowest negative free binding energy (ΔG binding) was found as −5.13 kcal/mol. In conclusion, all results are evaluated together, it has been determined that VGB has cytogenotoxic effects in vitro and binds to DNA in silico with significant free binding energy.

Assessment of hOGG1 Genetic Polymorphism (rs1052133) and DNA Damage in Radiation-Exposed Workers

The aim of this study was to assess the effect of radiation exposure, human 8-oxoguanine DNA N-glycosylase-1 (hOGG1) exon 7 genetic polymorphism and confounding factors on DNA damage response. Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) and alkaline Comet assay method were applied to determine the hOGG1 genetic polymorphisms and DNA damage response. A total of 80 participants were enrolled in this study, consisting of 40 radiation-exposed workers as a case group and 40 non-radiation workers as a control group. The genotypes frequencies for controls were Ser/Ser (35%), Ser/Cys (32.5%), and Cys/Cys (32.5%), with frequencies of alleles being 326Ser (0.52) and 326Cys (0.48), whereas the genotypes frequencies for radiation-exposed workers (cases group) were Ser/Ser (17.5%), Ser/Cys (57.5%), and Cys/Cys (25%), with frequencies of alleles being 326Ser (0.46) and 326Cys (0.54). The results indicated that DNA damage response were not significantly higher in the exposed workers than in controls (22.55 ± 6.02 versus 21.72 ± 7.14; P=0.58). The time of exposure has a significantly negative correlation with comet tail length value among radiation workers. In addition, it was found that the DNA damage response was strongly associated with age and time of exposure with a decrease of 0.6 percent (P-value: 0.008) and 0.58 percent (P-value: 0.009), respectively. Whereas gender, smoking habit, and equivalent dose were not correlated with DNA damage. The single-nucleotide polymorphism of hOGG1 exon 7 (rs1052133) demonstrated no association with the extent of DNA damage in radiation-exposed workers.

The Antibacterial and Larvicidal Potential of Bis-(2-Ethylhexyl) Phthalate from Lactiplantibacillus plantarum.

Lactic acid bacteria produce a variety of antibacterial and larvicidal metabolites, which could be used to cure diseases caused by pathogenic bacteria and to efficiently overcome issues regarding insecticide resistance. In the current study, the antibacterial and larvicidal potential of Bis-(2-ethylhexyl) phthalate isolated from Lactiplantibacillus plantarum BCH-1 has been evaluated. Bioactive compounds were extracted by ethyl acetate and were fractionated by gradient column chromatography from crude extract. Based on FT-IR analysis followed by GC-MS and ESI-MS/MS, the active compound was identified to be Bis-(2-ethylhexyl) phthalate. Antibacterial potential was evaluated by disk diffusion against E. coli (12.33 ± 0.56 mm inhibition zone) and S. aureus (5.66 ± 1.00 mm inhibition zone). Larvicidal potency was performed against Culex quinquefasciatus Say larvae, where Bis-(2-ethylhexyl) phthalate showed 100% mortality at 250 ppm after 72 h with LC50 of 67.03 ppm. Furthermore, after 72 h the acetylcholinesterase inhibition was observed as 29.00, 40.33, 53.00, 64.00, and 75.33 (%) at 50, 100, 150, 200, and 250 ppm, respectively. In comet assay, mean comet tail length (14.18 ± 0.28 μm), tail DNA percent damage (18.23 ± 0.06%), tail movement (14.68 ± 0.56 µm), comet length (20.62 ± 0.64 µm), head length (23.75 ± 0.27 µm), and head DNA percentage (39.19 ± 0.92%) were observed at 250 ppm as compared to the control. The current study for the first time describes the promising antibacterial and larvicidal potential of Bis-(2-ethylhexyl) phthalate from Lactiplantibacillus plantarum that would have potential pharmaceutical applications.

Effects of cysteamine supplementation on cryopreserved buffalo bull semen quality parameters

This work aimed to determine the effect of cysteamine (25, 50, 100 and 200 μM) incorporated during dilution on frozen thawed buffalo semen quality. Semen was collected twice weekly for 7 consecutive weeks from three Egyptian buffalo bulls using an artificial vagina. Semen samples were pooled and extended with a Tris-based extender, cooled, equilibrated and finally frozen in liquid nitrogen. The diluted semen was evaluated for motility, viability, morphology, plasma membrane and DNA integrity, in addition to oxidative stress and in vitro fertilizing capability. The post thaw motility and velocity parameters noticeably increased with different concentrations of cysteamine (mainly 100 μM) during different incubation periods. The post thaw sperm viability and normality significantly (p < 0.05) improved with concentrations of 50 and 100 μM. Plasma membrane integrity substantially increased at all concentrations of cysteamine. Cysteamine reduced alanine aminotransferase (at all concentrations), aspartate aminotransferase (at 25–100 μM), and creatine kinase (at 100 and 200 μM). Cysteamine at a concentration of 100 μM noticeably enhanced the total antioxidant capacity and glutathione peroxidase and decreased nitric oxide production. Cysteamine, at concentrations of 100 and 200 μM, increased the DNA intensity in the comet head (%) and decreased the DNA % in the comet tail. The comet tail length and moment substantially decreased at concentrations of 50–200 μM. Cysteamine did not affect the in vitro fertilizing capability of sperm. In conclusion, cysteamine incorporation (mainly at a concentration of 100 μM) in buffalo semen extender showed varying protective effects on different sperm parameters against cryo-damage; however, it did not affect the in vitro fertilizing capacity of sperm.

Ecogenotoxicological studies for an early toxicity screening and monitoring in Epinephalus chlorostigma and Scamberomorus commerson

The current research work was planned to investigate genotoxicity in fish to screen and monitor the aquatic ecosystem. Epinephalus chlorostigma (Hamour) was collected from the contaminated areas of “Tarut Island” (26.571°N 50.056°E) in the Arabian Gulf near Dammam, Saudi Arabia. DNA fragmentation was detected by Comet assay and Micronucleus assay. Heavy metals' water quality parameters and concentrations (Pb, Cr, Zn, Mn, Cu, Cd, Sn, and Hg) were extensively higher than the WHO permissible limits. They were more than enough to have adverse effects on fish health. The highest DNA fragmentation was observed in E. chlorostigma, indicating its most heightened sensitivity to pollution. E. chlorostigma showed comet head diameters 63.33 ± 2.2, 83.59 ± 3.38, and 66.28 ± 2.13px from S1-S3, respectively. E. chlorostigma erythrocytes showed comet tail lengths as 16.66 ± 1.65, 16.20 ± 1.63, and 19.07 ± 1.81px from S1-S3. DNA damage was found to be 19.14 ± 1.38, 16.38 ± 1.26, and 19.95 ± 1.33 % from S1-S3, respectively. The tail moment was observed as 6.46 ± 0.79, 4.72 ± 0.69, and 7.14 ± 1.08, while the olive moment was recorded as 5.31 ± 0.51, 5.14 ± 0.52, and 6.01 ± 0.49, respectively from S1-S3. The highest frequency for single micronucleus induction, double micronucleus induction, and nuclear abnormalities was observed in E. chlorostigma collected from the polluted site of the study area. This study proposes that this fish species and novel DNA damage assays could be the best tools for toxicity screening and monitoring water bodies.

Protective effect of green synthesized Selenium Nanoparticles against Doxorubicin induced multiple adverse effects in Swiss albino mice

AimsDoxorubicin (DOX) is a widely used drug against multiple cancers. However, its clinical Use is often restricted due to multiple adverse effects. Recently, Selenium Nanoparticles (SeNPs) are gaining attention due to their low toxicity and higher biocompatibility, making them attractive nanoparticles (NPs) in medical and pharmaceutical sciences. Therefore, the current study aimed to assess if our biosynthesized SeNP from the endophytic fungus Fusarium oxysporum conjugated with DOX could alleviate the DOX-induced adverse effects. Main methodsFor this purpose, we investigated various genotoxic, biochemical, histopathological, and immunohistochemical parameters and finally analyzed the metabolite profile by LC-MS/MS. Key findingsWe observed that DOX causes an increase in reactive oxygen and nitrogen species (ROS, RNS), 8-OHdG, and malondialdehyde (MDA), decreases antioxidant defense systems and reduces BCL-2 expression in cardiac tissue. In addition, a significant increase in DNA damage and alteration in the cytoarchitecture of the liver, kidney, and heart tissues was observed by Comet Tail Length and histopathological studies, respectively. Interestingly, the DOX-SeNP conjugate reduced ROS/RNS, 8-OHdG, and MDA levels in the liver, kidney, and heart tissues. It also restored the antioxidant enzymes and cytoarchitectures of the examined tissues, reduced genotoxicity, and increased the BCL-2 levels. Finally, metabolic profiling showed that DOX reduced the number of cardioprotective metabolites, which DOX-SeNP restored. SignificanceCollectively, the present results describe the protective effect of DOX-conjugated SeNP against DOX-induced toxicities. In conclusion, DOX-SeNP conjugate might be better for treating patients receiving DOX alone. However, it warrants further thorough investigation.

EVALUATION OF CYTOTOXIC AND GENOTOXIC EFFECTS OF SAXAGLIPTIN

Saxagliptin (SAX) is an oral drug that is a hypoglycemic (between an anti-diabetic agent) dipeptidyl peptidase-4 inhibitor. In this study, cytotoxic, genotoxic effects and DNA damage of SAX on human lymphocytes were investigated. For this purpose, Single Cell Gel Electrophoresis (SCGE), Micronucleus (MN) and Mitotic Index (MI) tests were used. Based on the daily doses of SAX, 0.017, 0.035, 0.07, 0.14 µg/mL concentrations used for the study. SAX significantly reduced the MI only at the highest concentration (0.14 µg/mL) for 24 hours, and 0.07- and 0.14-µg/mL concentrations for 48 hours. SAX did not cause a statistically significant change in MN frequency (except for concentration 0.14 µg/mL). In the SCGE test, a statistically significant increase of comet tail length was observed at 0.07- and 0.14-µg/mL concentrations. SAX did not cause a statistically significant change in comet tail moment and tail intensity (except for concentration 0.14 µg/mL). As a result, SAX caused statistically differences in the SCGE, MI and MN tests only at the highest concentrations that are not recommended commercial use (except for tail length, 0.035 µg/mL). When the results of all these studies are evaluated together, it can be said that SAX has no aneugenic, mutagenic and clastogenic effects at daily doses in in vitro studies on human lymphocytes.

Sequential evaluation of DNA damage in patients with head and neck carcinoma receiving radiotherapy

Background: Head and neck cancers account for about 30% of all cancers in India. Studies showed that there is an increased primary DNA damage even before the commencement of any modality of treatment in cancer patients which is further increased by the treatment. Chemo-radiation induced DNA damage is not repaired so effectively in patients with carcinoma which might pave way for secondary carcinoma. Aims and Objectives: The aim of this study was to assess the degree of DNA damage by comet assay technique in patients with head and neck carcinoma receiving radiotherapy. The degree of DNA damage was compared according to the age, gender, and associated risk factors of the patients. Materials and Methods: 35 patients with Stages II, III, and IVA, histopathologically confirmed Squamous cell carcinoma of head and neck with Karnofsky Performance Status &gt;70 attending radiotherapy OPD for treatment were included in this study.1 ml of heparinized blood was collected from the study participants during various doses of radiation treatment. All the samples were processed immediately and analyzed for DNA damage by single cell gel electrophoresis assay - Comet assay technique. Results: The comet length parameter, head diameter, and tail length were found to be increased when compared to baseline sample. The percentage of DNA in head parameter of post-RT sample was decreased when compared to baseline sample All these findings are indicative of DNA damage following radiotherapy. Conclusion: Patients with locally advanced head and neck carcinoma following radiotherapy showed a sequential increase in the DNA damage. The co-existing risk factors and old age may increase the baseline DNA damage in the patients with head and neck cancers.

Polycyclic aromatic hydrocarbons, long non-coding RNA expression, and DNA damage in coke oven workers.

Exposure to polycyclic aromatic hydrocarbons (PAHs) was associated with DNA damage, while the roles of long non-coding RNAs (lncRNAs) in the associations were unclear. We aimed to assess the association of lncRNA NR_024564 with urinary monohydroxy PAHs (OH-PAHs) and DNA damage among 332 coke oven workers. We determined 12 OH-PAHs by gas chromatography-mass spectrometry, and the expression level of NR_024564 by droplet digital RT-PCR and DNA damage by the comet assay. In total participants, we found that NR_024564 was not significantly associated with OH-PAHs or comet parameters. However, among workers with ≥ 20 working years, multiple OH-PAHs including urinary 1-hydroxyphenanthrene (1-OHPh), 2-OHPh, 3-OHPh, 9-OHPh, 1‑hydroxypyrene, and total PAH metabolites were related to increased comet parameters. Moreover, NR_024564 was significantly associated with 2-OHPh and four comet parameters. Each 1% increase in 2-OHPh was associated with 0.35% reduction (95% CI: 0.16%, 0.55%) in NR_024564 (P-FDR = 0.005), and 2-OHPh was marginally interacted with working years in relation to NR_024564 decrease. Also, each 1% increment of NR_024564 was related to 0.04-0.13% decrease of Olive tail moment, percent DNA in the comet tail, tail length, and tail moment (all P-FDR < 0.05). Furthermore, low NR_024564 level combined with high levels of 1-OHPh and 2-OHPh or ≥ 20 working years was positively associated with the comet parameters among the total participants. Our results indicated that NR_024564 might be linked to the adverse associations of PAHs with the DNA damage of coke oven workers who worked for ≥ 20years.

HPRT gene mutation, sister chromatid exchange and comet assay in peripheral lymphocytes of radiation laborers at Al-Amal Cancer Hospital in Baghdad

A Study performed on 30 Iraqi radiation Labors had been exposed tо a low dose tо ionizing radiation at Al-Amal Cancer Hospital in Baghdad the use of three genetic endpoints. Covered 12 females and 18 males between the aged tо (22-57) years. In addition to 20 Healthy individuals from the population, dwelling Baghdad, covered, 7 females and 13 males, aged (19 - 55) years which are non-smokers, non- alcoholic. The cytogenetic analysis, including, HPRT gene mutation¸ comet assay and SCE have been carried out on peripheral blood lymphocytes of labors and control groups. The HPRT gene mutation assay showed a significant increase (p&lt;0.05) in the radiation labors. There were found a significant increase (p&lt;0.05) in comet tail length and tail moment values in thе human lymphocyte in these radiation Labors compared with thе control group. Also the SCE in human lymphocyte for radiation Laborers was significantly (p&lt;0.05) higher than in a control group. Results obtained confirmed the cytogenetic analysis; including, HPRT gene mutation¸ comet assay and SCE are useful as biodosimetric markers for the detection of human exposure to low doses of ionizing radiation. It reflects the simultaneous exposure and actual levels of DNA damage present in radiological laborers' peripheral blood leukocytes by detecting temporary DNA damage and / or repair activity.