Olive Moment Research Articles

DNA damage in workers exposed to mineral oils

Mineral oils, untreated or mildly treated, have been classified in group 1 as a potential source of cancer by the International Agency for Research on Cancer (IARC). Although numerous studies have implicated metalworking fluids (MWFs) as human carcinogens, toxicology data regarding the mechanism of carcinogenicity are limited. This study is intended to examine the systemic effects of machining workers’ exposure to MWFs. The potential toxicity of mineral oils was investigated in 65 lathe workers compared to controls (66 men). The occupational exposure was measured by the National Institute for Occupational Safety and Health (NIOSH) 5026. The DNA damage has been examined by the comet assay method. According to the field assessments, the time-weighted average (TWA) exposure to mineral oil mist was 7.67 ± 3.21 mg/m3. A comet assay of peripheral blood cells showed that tail length (TL) and olive moment (OM) were significantly higher in the exposed group (p < 0.05). A multiple logistic regression analysis revealed that, within subjects with over 10 years of exposure, the odds ratio of worker with high TL, percent of DNA in tail, OM, and tail moment (TM) were 1.68, 1.41, 1.71, and 2.71, respectively. DNA strand break in exposed workers was associated with higher exposure time in years. Mineral oil toxicity could be altered in the presence of by-products and impurities. For a better understanding of genotoxicity, further studies are required.

Abstract 4810: HIF1α is involved in radiation-induced DNA damage repair and anti-apoptotic pathways in pancreatic ductal adenocarcinoma

Abstract Purpose/Objective(s): Pancreatic ductal adenocarcinoma (PDAC) is characterized by an unusually high degree of hypoxia in the tumor microenvironment, which activates hypoxic signaling pathways orchestrated by hypoxia-inducible factor 1α (HIF1α). It has been shown previously that HIF1α significantly increases PDAC radioresistance and growth, though its mechanism of action is unknown. Here, we test the hypothesis that HIF1α confers these traits in PDAC through involvement in DNA damage repair and KRAS signaling in vitro. Materials/Methods: We performed studies with the KPC cell line (KrasLSL-G12D/+;TP53LSL-R172H/+). We used CRISPR-Cas9, a genome editing tool, to generate a HIF1α knockout (KO) cell line. Hypoxic microenvironments were induced using 100uM of the oxygen scavenger CoCl2. Radioresistance and double-strand DNA damage was evaluated and compared between wild-type (WT) and KO cells after treatment with increasing doses of ionizing radiation using neutral comet assays. Tumor signaling pathways were mapped and analyzed using western blot and co-immunoprecipitation. The KRASG12D inhibitor, BI-2852, was obtained from Boehringer-Ingelheim. Cancer cells were evaluated for growth and migration using cell proliferation assays and scratch wound assays following treatment with 50 uM BI-2852. Bioinformatic analysis was conducted using TCGA (The Cancer Genome Atlas) data; high and low expression were defined as samples with mRNA expression z-scores ≥ 1 and ≤ -1, respectively, Results: HIF1α KO cells demonstrated significantly increased radioresistance compared to WT cells following 8 and 10 Gy of radiation in hypoxic conditions (t-test, P&amp;lt;0.05). The olive moment of KO cells treated with the IC50 radiation dose (4 Gy) was significantly higher than that of WT cells (t-test, P&amp;lt;0.0001), suggesting that the DNA damage repair was upregulated by HIF1α. In addition, WT cells demonstrated higher tumor proliferation (t-test, P&amp;lt;0.01) and migration (t-test, P&amp;lt;0.05) compared to KO cells under hypoxia, suggesting HIF1α promoted tumor growth. WT cells treated with BI-2852 demonstrated significantly lower levels of TP53; using coimmunoprecipitation, we show that TP53 was likely degraded through Mdm2. HIF1α KO was necessary to induce high levels of apoptosis in cells treated with BI-2852. Our results suggest that HIF1α may act synergistically with KRAS to promote tumor survival through anti-apoptotic pathways that target TP53 for degradation. TCGA data also suggest that tumors with high expression of HIF1α is are associated with significantly poorer prognosis coimpared to tumors with low expression of HIF1α (Logrank test, P=0.003). Conclusion: Overall, this study demonstrates that HIF1α promotes DNA damage repair in PDAC and reveals a HIF1α/KRAS/p53 signaling axis. Future studies will test anti-HIF1α and anti-KRAS combination therapies in PDAC mouse models. Citation Format: Kevin J. Tu, Sanjit K. Roy, Amit Sawant, Hem D. Shukla. HIF1α is involved in radiation-induced DNA damage repair and anti-apoptotic pathways in pancreatic ductal adenocarcinoma. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4810.

Dietary administration of probiotic Bacillus coagulans and Mentha piperita can protect histological architecture and DNA damage in Catla catla (Hamilton, 1822)

The present experimental investigation was executed to evaluate the combined effect of probiotic bacterium Bacillus coagulans along with herb Mentha piperita on histological architecture; and extent of DNA damage after challenge trial with pathogenic bacterium Aeromonas hydrophila in Catla catla. In this experiment, six dietary treatments in triplicate were formulated; C0, M3 and M6 in which M. piperita was supplemented at 0, 3, 6 g kg−1 of feed, while in dietary treatments CP, MP3 and MP6; supplementation of probiotic bacterium B. coagulans at the rate of 3000 CFU g−1 was done along with different inclusion level of M. piperita (0, 3, 6 g kg−1 of feed). Fishes were fed for 60 days on different dietary treatments. Growth performance, histological study and extent of DNA damage using comet assay was examined. Significantly (p < 0.05) high growth % gain in BW was observed in treatment MP6 followed by treatment M6. The histological analysis clearly revealed that dietary M. piperita without or with probiotic bacterium can protect the histoarchitecture of intestinal layers and liver even after challenge with A. hydrophila. Analysis of various comet parameters (tail length, olive moment, tail DNA, head DNA) affirmed that M. piperita supplemented diet can reduce the extent of DNA damage in fishes after challenge trial with A. hydrophila, however dietary supplementation of probiotic bacterium along with M. piperita can reduce the DNA damage up to larger extent confirming its combined protective effect of supplemented diets.

Clinical Investigation of Leukocyte DNA Damage in COVID-19 Patients

This prospective cross-sectional study aimed to evaluate leukocyte DNA damage in coronavirus disease (COVID-19) patients. In this study, 50 COVID-19-positive patients attending the Erzurum City Hospital Internal Medicine Outpatient Clinic and 42 control group patients were included. DNA damage was detected in living cells through leukocyte isolation in 50 COVID-19-positive patients using the comet assay method. DNA tail/head (olive) moments were evaluated and compared. White blood cells (WBC), red blood cells (RBC), hemoglobin (HGB), neutrophils (NEU), lymphocytes (LYM), eosinophils (EO), monocytes (MONO), basophils (BASO), platelets (PLT), and the neutrophil/lymphocyte ratio (NLR) were analyzed. The RBC, lymphocyte, eosinophil, and monocyte means were significantly higher in the control group (p < 0.05), whereas the HGB and neutrophile means were significantly higher in the study group (p < 0.05). There were significant negative correlations between COVID-19 and RBC (r = -0.863), LYM (r = -0.542), EO (r = -0.686), and MONO (r = -0.385). Meanwhile, there were significant positive correlations between COVID-19 and HGB (r = 0.863), NEU (r = 0.307), tail moment (r = 0.598), and olive moment (r = 0.582). Both the tail and olive moment mean differences were significantly higher in the study group, with higher ranges (p < 0.05). COVID-19 infection caused statistically significant increases in both the tail and olive damage percentage in patients, causing DNA damage. Lastly, the NLR rate was associated with the presence and progression of COVID-19.

Novel benzylidene benzofuranone analogues as potential anticancer agents: design, synthesis and in vitro evaluation based on CDK2 inhibition assays

The classical anticancer agents do not have their efficacy on inhibiting the G2 phase of the cell cycle. There are a very few reports available on drugs that work at G2 phase. Flavopiridol is one such drug candidate. In the current study, we sought to make analogues of flavopiridol. Still, the conditions used during their synthesis were unfavourable for the formation of flavopiridol and led to the generation of benzofuranones. In the present work, a new series of benzylidene benzofuranones were designed, synthesized and evaluated for their antioxidant, anti-colorectal cancer activity. Molecular docking, MMGBSA and molecular dynamics studies were conducted to assess their binding affinity at the active site of CDK2. Based on the cytotoxicity exhibited by test compounds, the compound NISOA4 (from isopropyl series) was further selected for mechanistic anticancer studies on HCT 116 cell lines. The compound selected was evaluated by comet assay, DNA fragmentation assay, cell cycle analysis, apoptosis detection by annexin FITC, semi-quantitative RTPCR based gene expression studies and FRET assay on the target CDK2/Cyclin A. Compound NISOA4 exhibited marked olive moments in comet assay studies. The apoptotic DNA fragmentation for compound NISOA4 demonstrated a marked change in the DNA fragmentation. The compound exhibited cell cycle arrest at G2/M phase at both the test concentrations. Apoptosis induction was observed at both the test concentrations and the compound was found to be a potent proapoptotic agent. It exhibited marked inhibition for the CDK2 gene expression and did not show any effect on CyclinA gene expression. However, the compound NISOA4 along with other analogues showed appreciable inhibition for the CDK2/Cyclin A target enzyme.

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.

Abstract P2058: LncRNA PCAT19 Limits Angiogenesis And Safeguards Quiescent Endothelial DNA By Preventing Uncontrolled Phosphorylation Of Replication Protein A2

Background: Long noncoding RNAs (lncRNA) provide an additional layer of molecular control in pathways crucial for vascular development and cardiovascular disease. We set out to identify novel endothelial lncRNAs that could be exploited to treat vascular disease. Methods and Results: We identified the endothelial cell-specific long noncoding RNA (lncRNA) PCAT19, which contributes to the endothelial proliferation-quiescence switch and acts as a safeguard for the endothelial genome during quiescence. PCAT19 was upregulated with endothelial cell confluence and quiescence. Knockdown of PCAT19 promoted endothelial proliferation and angiogenic sprouting. Conversely, PCAT19 overexpression reduced endothelial proliferative capacity and angiogenic sprouting. Using a cardiac organoid model system, we observed a markedly denser vascular network after the removal of PCAT19. RNA-sequencing after PCAT19 knockdown revealed multiple differentially regulated cell cycle genes. An antisense-oligonucleotide pulldown of PCAT19 followed by mass spectrometry identified multiple DNA damage response and cell cycle-related proteins as PCAT19 interaction partners; among them was the full DNA replication protein A complex. PCAT19 knockdown was found to sensitise DNA to damage, as measured by heightened p53 and γH2AX levels as well as a positive TUNEL signal and longer comet tail olive moments. Mechanistically, PCAT19 limited the phosphorylation of RPA2 on the serine 33 (S33) residue by inhibiting RPA2 interaction with the ATR kinase. Conclusions: In healthy vessels, endothelial cells maintain a stable, differentiated and growth-arrested phenotype. Upon injury, a rapid phenotypic switch facilitates proliferation to restore tissue perfusion. PCAT19 was identified as a highly enriched endothelial lncRNA that acts as a dynamic guardian of the endothelial genome, promotes cellular longevity and facilitates rapid switching to proliferation. Therapeutic targeting of PCAT19 could potentially alter the course of vascular disease development and progression by modulating endothelial growth and angiogenic sprouting.

Abstract 1630: Development of a clinically relevant in vitro model of chemo-radioresistant cervical cancer cells for insights into treatment resistance

Abstract Resistance to therapeutic treatment is a common phenomenon that contributes to relapse in several cancers including cervical that is the fourth most common cancer in women across the world. Acquired resistance to chemotherapy, radiotherapy or their combination contributes to a majority of relapse cases which suggests the need to develop models that can be used for preliminary screening of drugs or decipher the associated signaling mechanisms contributing to this resistance. Various chemo-resistant, as well as radio-resistant models, have been established as in vitro models. In cervical cancer treatment, often, radiation therapy is accompanied by the addition of radio-sensitizers such as cisplatin that increase its treatment efficiency. Hence, a combined chemo-radio-resistant in vitro model seems more relevant clinically in cervical cancer. Therefore, we designed our study to establish a cellular model that is resistant to a combined treatment of chemotherapy and radiotherapy. A chemo-radio resistant cervical cancer cell line (HeLa) was developed in vitro by a fractionated exposure of a combination of cisplatin and megavoltage X-rays. Briefly, HeLa cells were sensitized with cisplatin (0.1µM increase at every fraction) prior to radiation exposure (2Gy). The cells were treated once in 5 days and eventually after 25 fractions, delivered in 20 weeks, the cells were made resistant up to a total dose of 2.5µM cisplatin + 50Gy. These cells were further characterized and compared with radioresistant HeLa cells as well as parental cells. For this purpose, proliferation assay, clonogenic survival assay, cell cycle analysis and apoptosis assay were performed. The resistivity was confirmed by comet assay and Gamma-H2AX staining. Interestingly, an increase in cell proliferation and clonogenic survival rates was observed in chemo-radio-resistant cells. FACS analysis revealed that the proportion of cells in G2/M phase increased and apoptotic rates declined in chemo-radio-resistant cells. Further, changes in comet tails, olive moment and Gamma-H2AX foci formation in chemo-radio-resistant groups confirmed resistivity. These cells were also analyzed for their metastatic properties and results suggest that induced chemo-radio-resistant cells are more tumorigenic as compared to only radioresistant cells as well as the parental ones. In conclusion, the establishment of a chemo-radio-resistant cell line may serve as a model to study signalling mechanisms, drug screening and develop novel drugs. Further studies are underway to induce the chemo-radio resistance in other cell lines by a similar method. Citation Format: Sanchita Khurana, Surbhi Singla, Reena Sharma, Bhavana Rai, Shalmoli Bhattacharyya. Development of a clinically relevant in vitro model of chemo-radioresistant cervical cancer cells for insights into treatment resistance [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1630.

DNA damage in hepatopancreas cells of Haliotis discus hannai induced by P-xylene

In order to examine the toxic effects of p-xylene on the hepatopancreas of Haliotis discus hannai, a subchronic toxicity test was carried out for 21 days by setting 4 concentrations (0.5, 1.0, 1.5, and 2.0 mg·L-1) and the control group. DNA damage in hepatopancreas cells of H. discus hannai was analyzed by comet assay. The CASP analysis software was used to analyze the damage indices, including tail rate, tail length, percentage of tail DNA, and olive moment. The results showed that DNA damage in hepatopancreas cells occured in all exposed groups, with different degrees of damage. The DNA damage of hepatopancreas cells was aggravated with the increase of exposure concentration. High concentration of p-xylene could induce apoptosis, showing a certain dose-damage effect. Short-term exposure to medium concentration of p-xylene could cause DNA damage in hepatopancreas cells of H. discus hannai, and the damage degree increased with the extension of exposure time, showing a certain time-damage effect. However, the damage indices of DNA decreased after prolonged exposure, which might be related to the DNA repair mechanism of the cell and the metabolic mechanisms of the detoxification system. In conclusion, p-xylene could induce oxidative damage in hepatopancreas cells of H. discus hannai, resulting in DNA fragmentation. Prolonged exposure to high concentration of p-xylene could lead to apoptosis.

Genotoxic impact of di-n-butyl phthalate on DNA: A comparative study of three generations in the neuronal tissue of Wistar rats.

Di-n-butyl phthalate (DBP), one of the plasticizers, is considered a ubiquitous environmental contaminant due to its widespread application in personal-care products and serves as a raw material in many industries for the generation of many plastic products. Several scientific investigations have shown that DBP caused embryotoxicity and cognitive impairments. However, there is less understanding of the genotoxic potential of DBP in neuronal tissue when exposure happens continuously for several generations. The present study was undertaken to investigate the impact of DBP on the nucleic acids of neuronal tissue in one-month-old rats by performing a comet assay and biochemical analyses. By oral gavage, the parental generation (F0) was administered DBP (500mg/kg/day) during gestation (GD6-20) and lactation, and exposures were continued for three consecutive generations until the pups were grown to one-month-old. The oxidative stress assessments carried out in discrete brain regions isolated from one-month-old rats (F1-F3) following DBP exposure indicated significant inhibition in the levels of antioxidant enzymes (superoxide dismutase and catalase) while oxidant status (malondialdehyde) was elevated significantly. The extent of DNA damage using the comet assay, as measured by the olive moment, tail DNA percentage and tail length, was greater in DBP-treated rats compared with the control group, but RNA/DNA content decreased significantly. The results of this study suggested a strong link between oxidative stress and genetic integrity in the neuronal tissue of rats exposed to DBP generationally. To summarise, DBP exposure during pregnancy caused oxidative stress, which resulted in genetic instability in specific discrete brain regions of the third generation.

EFFECT OF LUTEIN INTERVENTION IN ALLEVIATING THE POST-RADIATION EFFECTS OF ELECTRON BEAM RADIATION

Ionizing radiations are harmful since they generate free radicals which through a series of reactions may cause damage to genetic material, imbalance in the antioxidant system, or lead to death. Lutein; a carotenoid compound because of its various medicinal properties has been chosen to evaluate its protection against radiation-induced damages. Swiss albino mice were divided into 8 groups of 6 mice each. Irradiation groups received whole-body radiation of 6Gy using an electron beam accelerator. Mice were fed with water/gallic acid/10% dimethyl sulfoxide (DMSO) (w/v)/ lutein respectively. The mice were sacrificed on the 16th day; whole blood was collected in 2% ethylene diamine tetraacetic acid (EDTA) tubes by cardiac puncture method for hematological studies and comet assay. The organs like the brain, lungs, and liver were dissected. Organ homogenates were prepared to perform the antioxidant assay. The femur of the leg was removed and flushed for micronucleus assay. Lutein post-treatment showed significantly increased superoxide dismutase activity in the lung homogenate in comparison to the radiation control. A significant increase in the levels of glutathione was observed in lutein post-treatment when compared to the lutein control in lung homogenate. Also in the lung homogenate of lutein post-treatment, a significant decrease in the levels of malondialdehyde was observed in comparison to the lutein control, and a similar effect was observed in the vehicle control groups. In the comet assay, it was observed that tail moment decreased significantly in lutein post-treatment when compared to its control group whereas no significant changes were observed with %DNA in tail and olive moment. Lutein treatment post-radiation has increased the polychromatic erythrocytes (PCE)/ (PCE+ normochromic erythrocytes (NCE) ratio. Significant changes were observed in the lutein post-treatment group with respect to antioxidant status. Micronucleus assay reveals that lutein treatment post-radiation increases cell multiplication. These results indicate a potent mitigator effect of lutein against radiation-induced antioxidant changes in vivo.

Genotoxic and histopathological alterations in rats exposed to herbal liquors

The increase in consumption of herbal liquors in Nigeria is a cause for alarm. These are consumed with the mis-conception that they are without toxic effects. The aim of this study is to investigate the genotoxic and histopathological alterations in rats exposed to herbal liquors. Female rats were exposed to herbal liquors for 6 weeks. The histopathological and genotoxic evaluation were done to assess extent of damage. Pathological examination revealed incidences of aggregates chronic inflammatory cell infiltrates in the heart of Jedi treated group while the heart of the other groups had no abnormalities. The histologic sections of the kidney tissue revealed congested vessels while the lung showed reduction in air filled alveolar spaces with infiltration of alveoli and interstitium by aggregates of inflammatory cells indnadicating moderate to severe pulmonary inflammation. Histologic sections of lung tissue in rats treated with herbal liquors reveals congestion of pulmonary vessels and interstitial hemorrhages. Genotoxic evaluation of rat lymphocytes exposed to herbal liquors via comet assay shows that rats administered with the different herbal liquors developed significant (p &lt; 0.05) as revealed in the % DNA in tail, % DNA in head, olive moment, tail length and tail moment which indicates the presence of DNA strand breaks and a marker for oxidative DNA damage. This result reveals that herbal liquors contain substances that produce reactive oxygen species that have pathological effect on certain organs as well as inducing DNA strand breaks that could compromise the integrity of the DNA which can lead to mutation.

Effect of occupational exposure to antineoplastic drugs on DNA damage in nurses: a cross-sectional study

BackgroundAlthough the therapeutic effect of antineoplastic drugs is incontestable, these agents can also potentially act as carcinogens, mutagens and/or teratogens in people. The aim of this study was to assess...

Association of DNA damage with vitamin D and hair heavy metals of obese women

BackgroundObesity has been linked to DNA damage. The modifiable risk factors may modulate the impact of obesity on DNA damage.ObjectiveThis study aimed to assess DNA damage and its association with dietary nutrient, serum 25-hydroxyvitamin D (25(OH)D) and concentration of hair heavy metals of obese and non-obese women.MethodA case–control study was conducted involving 134 women aged between 20 and 50 years. Serum 25(OH)D, fasting glucose, and lipid profile were assessed. Indicators of DNA damage such as percentage of tail DNA, tail moment, tail olive moment, tail intensity and tail length were measured using an alkaline-comet assay. Concentrations of hair heavy metals were quantified using inductively coupled plasma–mass spectrometry (ICP–MS). Participants' daily energy, macro, and micronutrient intake were collected using the Food Frequency Questionnaire.ResultsMean values of serum 25(OH)D was 31.8 ± 0.9 nmol/L. 96.3% of participants were vitamin D deficiency (< 50 nmol/L). The mean BMI was 26.3 ± 0.5 kg/m2. Half of the participants (50.7%) have a high frequency of DNA strand breaks. Mean concentration of hair heavy metals (mg/kg) were 0.1 ± 0.03 (arsenic), 0.2 ± 0.1 (cadmium), 1.0 ± 0.4 (mercury), 2.8 ± 0.8 (lead),and 6.2 ± 0.4 (chromium). There was no significant difference for the mean of serum 25(OH)D, indicators of DNA damage, concentrations of hair heavy metals and dietary nutrients between obese and non-obese groups (p > 0.05). Obese women with serum 25(OH)D level of ≥ 31 nmol/L had a significantly lower tail moment (p = 0.029) and tail olive moment (p = 0.031); thus, indicating less DNA damage. Additionally, obese women with hair chromium concentration of ≥ 5.88 mg/kg had a significantly higher tail moment (p = 0.047), indicating more DNA damage.ConclusionDNA damage among obese women correlated with serum 25(OH)D and hair chromium.

In Vivo Genotoxicity Assessment of Gold Nanoparticles of Different Doses by Comet Assays

Gold nanoparticles were synthesis by green method using the soaked orange peels (citrus Sinensis) as reducing and stabilizing agent. The gold nanoparticles were diagnosed using the approved method, including the transmission electron microscope and the visible UV spectroscopy. Thus, the results of the diagnosis showed the formation of spherical particles with a size of 36 nanometers. Their biological effects on the bone marrow cells of albino mice were studied. Male albino mice were used in this study and randomly divided into seven groups. The first group was the control group injected with the physiological solution (Normal Saline) and the other six groups were injected with different doses of the solution of the gold nanoparticles (1, 2, 4, 6, 8 and 10) Mg/kg. The effect of gold nanoparticles on the DNA damage by bone marrow cells was studied using comet assay according to the following criteria: (DNA in Head%, Comet Length, Tail Length, DNA in Tail%, Olive Moment), the results showed a significant decrease (P?0.01) in the rate of DNA in Head%, while it showed a significant increase in Comet Length, Tail Length, DNA in Tail%, Olive Moment compared to the control group and all Doses studied. The effect of gold nanoparticles was directly proportional to the increase in the studied doses. The dose was 10 mg/kg, which was the highest toxicity, followed by the dose of 8 mg/kg, then the rest of the doses was descending.

Cytotoxicity and genotoxicity of gold nanorods assisted photothermal therapy against Ehrlich carcinoma in-vivo

AimPreparation of pegylated gold nanorods (PEG-AuNRs) that are capable of converting near infrared (NIR) light into heat. Evaluation of cancer therapeutic efficacy and long-term toxicity of the proposed photothermal therapy in comparison with other conventional modalities. Materials and methodsPrepared PEG-AuNRs were characterized by measuring their absorption spectra, zeta potential, and transmission electron microscope (TEM). Cancer therapeutic efficacy was assessed by monitoring tumor growth, measuring DNA damage and superoxide dismutase (SOD) and malondialdehyde (MDA) levels in addition to examining tumor histopathology. Further analysis concerning the toxicity of all the proposed treatment modalities was also assessed by evaluating the cytotoxicity and genotoxicity in liver and kidney tissues. Key findingsThe results demonstrated that both photothermal therapy (PEG-AuNRs + NIR laser) and chemotherapy (cisplatin) have higher efficacy in diminishing Ehrlich tumor growth with significance DNA damage over the other treatment modalities.Concerning the biosafety issue, mice treated photothermally exhibited lower MDA level and higher SOD activity in liver and kidney tissues compared with other treated groups. DNA damage represented by tail moment and olive moment of kidney tissues exhibited lower values for photothermal treated group and higher values for cisplatin treated group. SignificancePhotothermal therapy (PEG-AuNRs + NIR laser) potentiates higher efficacy in treating Ehrlich tumor with minimum toxicity in comparison with other conventional treatment modalities.

Assessment of the genotoxic potential of a migraine-specific drug by comet and cytokinesis-block micronucleus assays

ABSTRACTBackground: Eletriptan is a migraine-specific drug-containing the triptan group. In terms of drug safety, the present study aimed to investigate the genotoxic potential of eletriptan.Research design & methods: We conducted our study by using the cytokinesis-block micronucleus cytome (CBMN) assay, a comprehensive method for measuring micronucleus formation, and a sensitive method for detecting DNA-strand breaks. In the assay, cytokinesis-block proliferation index and the frequency of micronuclei were evaluated in lymphocytes treated with three different concentrations (1, 10 and 25 µg/ml) of eletriptan for 48 hours. In comet assays, DNA damage was evaluated in leucocytes treated with three different concentrations (1, 10 and 25 µg/ml) of eletriptan for an hour.Results: Eletriptan did not induce cytotoxicity nor any increased micronuclei frequencies. While the comet parameters % DNA in tail, tail moment, and the olive moment was found to be significantly increased at 10 and 25 µg/ml, the cytokinesis-block proliferation index values were not.Conclusion: These findings suggest that eletriptan is non-cytotoxic but potentially weakly genotoxic at higher concentrations (10 and 25 µg/ml).

Lymphocyte DNA damage in children with iron-deficiency anemia: a case–control study

Context Iron-deficiency anemia (IDA) is a widespread yet one of the most neglected micronutrient deficiency disorder worldwide. Aims The objective was to evaluate lymphocyte DNA damage in children with anemia. Materials and methods This was a prospective case–control study. A total of 80 infants and children aged six months to twelve years were included. Fifty had IDA, whereas 30 were controls. DNA damage scoring using alkaline comet assay was done in all children. Percentage and mean and/or SEM were calculated. Comparison between cases and controls was done using Student’s t test. Analysis of variance test was used to compare the groups within the cases and controls. Pearson correlation coefficient was performed to relate the hemoglobin levels with the DNA damage. Statistical software was used for analysis. Results DNA damage scoring was significant in all the parameters in the iron-deficiency group with respect to the tail length and percentages of DNA in tail and olive moment, with a P value of 0.006, 0.002, and 0.038, respectively. A statistically significant negative correlation was found between hemoglobin levels and percentage of DNA in tail (r=−0.280; P=0.012) as well as olive moment (r=−0.240; P=0.032) Conclusion IDA was associated significantly with lymphocyte DNA damage. A significant negative correlation between hemoglobin levels and percentage of DNA and olive moment was also elucidated. The authors conclude that early intervention is needed in even mild cases of IDA.

A comparative evaluation of the DNA damage in the serum of chronic periodontitis patients with and without diabetes mellitus type II.

Periodontal disease is an immunoinflammatory disease that is initiated by the interaction between microbial plaque and the periodontal tissues. There is very limited data available on the assessment of DNA damage with relation to periodontal diseases. Therefore, a need for a study in this area was felt. To evaluate the DNA damage in the serum of chronic periodontitis patients and chronic periodontitis with diabetes mellitus (DM) type II patients and to compare it with healthy controls, to assess whether periodontitis can have systemic effects beyond the periodontium. This cross-sectional study was conducted involving 150 subjects in the age group of 30-60 years, from October 2010 to May 2015. A blood sample of 5 ml of venous blood was collected from each of the study subjects, from the antecubital vein. Fresh blood was used to assess the DNA damage. The DNA damage was estimated using the alkaline single-cell gel (comet) assay. The DNA damage to the cells was calculated by assessing the percentage of "DNA in tail." The results showed that the values were higher in the periodontitis with diabetes group, as compared to the periodontitis and control group. When the Olive moment was calculated, the values were higher in the periodontitis with diabetes group as compared with the other two groups. Although the values were seen to be higher, it was not statistically significant. The results obtained from this study although statistically insignificant suggest that the DNA damage was higher in chronic periodontitis as compared with healthy control. There was a potentiated difference of the values in patients with DM type II when compared to chronic periodontitis alone.

Anticancer activity of Juniperus procera grown in southwestern region of Saudi Arabia on human oral squamous cell carcinoma cell lines

Background: One of the popular ancient plants used to treat various diseases such as hyperglycemia, hepatitis, jaundice, bronchitis, and pneumonia has been Juniperus procera (JP). JP is abundantly seen in the region of Al-Baha, Saudi Arabia, and is being used as a medicinal plant traditionally by local healers. Objectives: The objective was to evaluate the anticancer properties of JP from Al-Baha region on SCC-9 cells. Materials and Methods: Colorimetric assays such as 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, sulforhodamine B assay, and lactate dehydrogenase (LDH) assay were performed to check the cytotoxicity induced and cell viability. Assays evaluating cellular events such as apoptosis and cell cycle were also performed. Results: MTT assay revealed IC50value of 201.6 μg/ml. The cancer cells primed with increasing concentrations of JP displayed an enhanced emission of LDH at elevated concentrations contrasted to cells which were not treated. The samples of JP pooled fraction (PF) (6–9) treated at 160 μg/ml and 320 μg/ml concentration showed 8.39% and 23.37% and 17.35% and 20.89% in early and late phases of apoptosis, respectively. Sample P (PF 6-9) at 160 μg/ml and 320 μg/ml has induced a G2M phase arrest of up to 21.06% and 26.94%. Deoxyribose nucleic acid damage was compared in tested concentrations of sample JP with untreated control cells in SCC9 cells. SCC9 cells that were treated with sample JP PF 6–9 showed the olive moments 23.22 and 37.30 at concentrations 160 μg/ml and 320 μg/ml, respectively. Studies of gene expression showed that increased concentrations of JP triggered the development of caspases and p53. Conclusion: The bioactive compounds found in JP were effective and potent against the SCC-9 cancer cells.