scholarly journals Assessment of DNA Damage in Leukoplakia Patients with Different Degrees of Dysplasia

2015 ◽  
Vol 16 (12) ◽  
pp. 971-976 ◽  
Author(s):  
Mohamed I Hashem ◽  
Zeeshan H Ahmad ◽  
Mohammed A Binmgren ◽  
Sukumaran Anil ◽  
Sahar Bin Huraib

ABSTRACT Background Single cell gel electrophoresis (SCGE) assay also known as comet assay is a rapid and highly sensitive fluorescent molecular technique for detecting various forms of deoxyribonucleic acid (DNA) damage at individual cellular level. Materials and methods The present study was done to detect the extent of DNA damage in oral leukoplakia (OL) and compare with normal individuals. The sample population was obtained from an outpatient clinic of a tertiary teaching dental institute. A total of 36 consecutive patients with leukoplakia and 10 healthy normal volunteers were recruited for the study and assessed for the extent of DNA damage using SCGE following clinical diagnosis and histological grading. Peripheral blood was obtained by venipuncture and SCGE assay was performed. Mean comet tail length was recorded and analyzed statistically to compare the extent of damage in each group. Results The mean comet tail length seen in leukoplakia patients with moderate to severe dysplasia was 1.25 ± 0.14 mm while for the control subjects, it was 0.31 ± 0.10 mm. The difference was statistically significant (p = 0.000). On comparing within the grades of leukoplakia, a progressive trend of increasing tail length was observed with increasing grades of dysplasia. Conclusion Deoxyribonucleic acid damage as measured by SCGE is seen in leukoplakia. A stepwise increase in DNA damage levels from healthy controls, through patients with non-dysplastic epithelium to varying grades of dysplasia has been observed indicating the extent of DNA damage in this high risk group. How to cite this article Vellappally S, Binmgren MA, Huraib SB, Hashem MI, Patil S, Anil S. Assessment of DNA Damage in Leukoplakia Patients with Different Degrees of Dysplasia. J Contemp Dent Pract 2015;16(12):971-976.

2018 ◽  
Vol 58 (2) ◽  
pp. 252 ◽  
Author(s):  
L. Fraser ◽  
Ł. Zasiadczyk ◽  
C. S. Pareek

Assessment of sperm-DNA integrity is a crucial issue in male fertility. In the present study, parameters derived from the image analysis of comets after single-cell gel electrophoresis were used to analyse the types of DNA damage of frozen–thawed boar spermatozoa. Semen, frozen in a cryoprotectant-free extender or in cryoprotectant-based extenders, was analysed for DNA fragmentation and with the following comet tail measures: percentage DNA in comet tail, comet tail length and olive tail moment. The percentages of sperm DNA damage in the comet tails were classified as Type 0 (no DNA damage), Type I (very low DNA damage), Type II (light DNA damage), Type III (medium DNA damage) and Type IV (heavy DNA damage). Sperm motility characteristics and membrane integrity were assessed in the pre-freeze and frozen–thawed semen samples. Assessment of sperm DNA fragmentation and comet tail measures showed marked inter-boar variability following cryopreservation. However, consistent differences among the boars, with respect to cryo-induced sperm DNA damage, were detected by the comet tail length and olive tail moment. Besides Type IV, all types of DNA damage were detected in the cryoprotectant-based extenders. It was found that the frequency of Type II and Type III of DNA damage of frozen–thawed spermatozoa was significantly greater in the cryoprotectant-based and cryoprotectant-free extenders respectively. Deterioration in the quality of the sperm DNA integrity was concomitant with a marked decline in sperm motility characteristics, reduced plasma membrane integrity and higher lipid peroxidation and aspartate aminotransferase activity after cryopreservation. It can be suggested that the comet-assay parameters, coupled with routine laboratory tests, are useful to improve the sperm evaluations of post-thaw quality of semen from individual boars and would offer more comprehensive information for a better understanding of the degree of cryo-induced sperm-DNA damage.


2021 ◽  
Vol 8 (1) ◽  
pp. 105-113
Author(s):  
Darlina Yusuf ◽  
Devita Tetriana ◽  
Tur Rahardjo ◽  
Teja Kisnanto ◽  
Yanti Lusiyanti ◽  
...  

Analyses of DNA Damage in the Patient’s Lymphocyte Cells Post-Radiotherapy Radiotherapy given in high doses to kill cancer cells can also induce DNA damage in surrounding normal cells. The radiation dose is divided into smaller doses called fractionation to decrease the effect of radiation on normal tissue. For this reason, it is necessary to monitor the peripheral blood lymphocytes to evaluate the patient's DNA damage. The alkaline comet test is a simple and sensitive technique for detecting DNA instability. This study involved 11 patients who underwent radiotherapy up to 20 Gy, and 11 healthy subjects as controls. This study aims to see how much DNA damage is caused by a 20 Gy fractionated radiation dose in patients with various cancers. The results showed that the mean frequency of damaged cells in patients was 80.54 ± 12.52% with a mean comet tail length of 49.98 ± 12.93 µm. There was a significant difference in both the frequency of damaged cells and the mean value of the comet tail length against the control group (p < 0.001). It was concluded that high doses of radiation can cause DNA damage to peripheral blood lymphocytes. Radioterapi yang diberikan dalam dosis tinggi untuk mematikan sel kanker juga dapat menginduksi kerusakan DNA pada sel normal di sekitarnya. Dosis radiasi dibagi menjadi dosis yang lebih kecil yang disebut fraksinasi untuk menurunkan efek radiasi pada jaringan normal. Untuk itu perlu pemantauan pada limfosit darah tepi untuk mengevaluasi kerusakan DNA pasien. Uji komet alkali merupakan teknik yang sederhana dan sensitif untuk mendeteksi ketidakstabilan DNA. Penelitian ini melibatkan 11 pasien yang menjalani radioterapi hingga 20 Gy, dan 11 subyek sehat sebagai kontrol. Penelitian ini bertujuan untuk melihat seberapa besar kerusakan DNA akibat dosis radiasi fraksinasi 20 Gy pada pasien dengan variasi kanker. Hasil penelitian menunjukkan bahwa rerata frekuensi sel yang rusak pada pasien 80,54 ± 12,52% dengan rerata panjang ekor komet 49,98 ± 12,93 µm terdapat perbedaan nyata baik pada frekuensi sel yang rusak maupun nilai rerata panjang ekor komet terhadap kelompok kontrol (p < 0,001). Penelitian ini menyimpulkan bahwa radiasi dosis tinggi dapat menyebabkan kerusakan DNA sel limfosit darah tepi.


2013 ◽  
Vol 33 (3) ◽  
pp. 224-229 ◽  
Author(s):  
T Zhang ◽  
Q Zhao ◽  
Y Zhang ◽  
J Ning

The present study investigated the genotoxic effects of flumorph in various organs (brain, liver, spleen, kidney and sperm) of mice. The DNA damage, measured as comet tail length (µm), was determined using the alkaline comet assay. The comet assay is a sensitive assay for the detection of genotoxicity caused by flumorph using mice as a model. Statistically significant increases in comet assay for both dose-dependent and duration-dependent DNA damage were observed in all the organs assessed. The organs exhibited the maximum DNA damage in 96 h at 54 mg/kg body weight. Brain showed maximum DNA damage followed by spleen > kidney > liver > sperm. Our data demonstrated that flumorph had induced systemic genotoxicity in mammals as it caused DNA damage in all tested vital organs, especially in brain and spleen.


2014 ◽  
Vol 522-524 ◽  
pp. 251-256
Author(s):  
Jun Song Han ◽  
Xin Lu Lv ◽  
Ya Li Gao ◽  
Xiang Gao ◽  
De Qi Xiong

The single cell gel electrophoresis (SCGE) is a rapid and sensitive procedure for measuring strand breaks in DNA. In the present study, sea urchin (Hemicentrotus pulcherrimus) was chosen as the test organism and SCGE was applied to assess DNA damage of its gonadal cells exposed to petroleum hydrocarbon. The gonadal cells of sea urchin had been seriously damaged above 50 mg/L of Water Accommodated Fractions (WAFs), whileas no damages occurred in the lower concentrations. There were good linear relationships between exposure days and DNA damage rate, percentage of DNA in the comet tail (%TDNA) as well as comet tail length (TL).


2011 ◽  
Vol 2011 ◽  
pp. 1-7 ◽  
Author(s):  
Towseef Mohsin Bhat ◽  
M. Y. K. Ansari ◽  
Sana Choudhary ◽  
Rumana Aslam ◽  
Alka

Coexposure to heavy metals occurs in many occupational settings, such as automobile refining shops, pigment, and batteries production. Heavy metals around automobile refining shops were tested for their ability to induce synergistic cytogenetic effects in Trifolium repens L. by using the chromosomal aberrasions (CAs), micronucleus (MN) and comet assay. A significant increase in micronucleus (MN), chromosomal abrations (CAs), percentage of nuclei with comet tails (NCTs), the relative comet tail length (CTL), comet tail DNA (CT, DNA), and tail moment (TM) were observed with increased concentration of three heavy metals, like Cd, Pb, Hg. The present result indicate that exposure of T. repens to soils contaminated by heavy metals around automobile refining shops shows clastogenicity, cytotoxicity, and DNA damage at higher concentrations.


2010 ◽  
Vol 30 (9) ◽  
pp. 1297-1302
Author(s):  
Tao Zhang ◽  
Jiye Hu ◽  
Yuchao Zhang ◽  
Qianfei Zhao ◽  
Jun Ning

JS-118 is an extensively used insecticide in China. The present study investigated the genotoxic effect of JS-118 on whole blood at 24, 48, 72 and 96 h by using alkaline comet assay. Male Kunming mice were given 6.25, 12.5, 25, 50 and 100 mg/kg BW of JS-118 intraperitoneally. A statistically significant increase in all comet parameters indicating DNA damage was observed at 24 h post-treatment ( p < 0.05). A clear concentration-dependent increase of DNA damage was revealed as evident by the OTM (arbitrary units), tail length (µm) and tail DNA (%). From 48 h post-treatment, a gradual decrease in mean comet parameters was noted. By 96 h of post-treatment, the mean comet tail length reached control levels indicating repair of damaged DNA. This study on mice showed different DNA damage depending on the concentration of JS-118 and the period of treatment. The present study provided further information of the potential risk of the genetic damage caused by JS-118.


Author(s):  
Hossam Ebaid ◽  
Mohamed Habila ◽  
Iftekhar Hassan ◽  
Jameel Al-Tamimi ◽  
Mohamed S. Omar ◽  
...  

Background: Hepatotoxicity remains an important clinical challenge. Hepatotoxicity observed in response to toxins and hazardous chemicals may be alleviated by delivery of the curcumin in silver nanoparticles (AgNPs-curcumin). In this study, we examined the impact of AgNPs-curcumin in a mouse model of carbon tetrachloride (CCl4)-induced hepatic injury. Methods: Male C57BL/6 mice were divided into three groups (n=8 per group). Mice in group 1 were treated with vehicle control alone, while mice in Group 2 received a single intraperitoneal injection of 1 ml/kg CCl4 in liquid paraffin (1:1 v/v). Mice in group 3 were treated with 2.5 mg/kg AgNPs-curcumin twice per week for three weeks after the CCl4 challenge. Results: Administration of CCL4 resulted in oxidative dysregulation, including significant reductions in reduced glutathione and concomitant elevations in the level of malondialdehyde (MDA). CCL4 challenge also resulted in elevated levels of serum aspartate transaminase (AST) and alanine transaminase (ALT); these findings were associated with the destruction of hepatic tissues. Treatment with AgNPs-curcumin prevented oxidative imbalance, hepatic dysfunction, and tissue destruction. A comet assay revealed that CCl4 challenge resulted in significant DNA damage as documented by a 70% increase in nuclear DNA tail-length; treatment with AgNPs-curcumin inhibited the CCL4-mediated increase in nuclear DNA tail-length by 34%. Conclusion: Administration of AgNPs-curcumin resulted in significant antioxidant activity in vivo. This agent has the potential to prevent the hepatic tissue destruction and DNA damage that results from direct exposure to CCL4.


2017 ◽  
Vol 37 (9) ◽  
pp. 901-908 ◽  
Author(s):  
AM Samir ◽  
LA Rashed

Aim: The aim of this work was to investigate the relationships between aluminium levels, oxidative status and DNA damage in workers occupationally exposed to aluminium. Subjects and methods: This study was conducted in a secondary aluminium smelter. It included 96 male workers occupationally exposed to aluminium fume and dust compared to 96 male nonexposed individuals. Full history and clinical examination were done for all participants. Laboratory investigations in the form of serum aluminium, total antioxidant capacity (TAC), urinary 8-hydroxy-2′-deoxyguanosine (8-OHdG) and comet assay test were performed. Results: Serum aluminium level ranged from 4 to 30 µg/L of median: 10 µg/L; urinary 8-OHdG ranged from 2.7 to 17.2 ng/mg creatinine of median: 7.6 ng/mg creatinine; comet tail length (CTL) ranged from 19.7 to 50.5 µm of median: 45 µm, were statistically significantly increased in the exposed group compared to nonexposed group. In exposed workers, a statistically significant positive correlations were found between serum aluminium level and urinary 8-OHdG ( r = 0.75, p < 0.001); aluminium level and CTL ( r = 0.71, p < 0.001); and urinary 8-OHdG and CTL ( r = 0.71, p < 0.001). There was a statistically significant negative correlation between serum aluminium and TAC ( r = −0.76, p < 0.001). Conclusion: Occupational exposure to aluminium in secondary aluminium smelters was related to the induction of oxidative stress and DNA damage. This may promote the development of adverse health hazards in the exposed workers


2019 ◽  
Vol 2 (1) ◽  
pp. 39-47
Author(s):  
Faiza Ambreen ◽  
Muhammad Javed

The present study was undertaken to examine the DNA damage in peripheral blood erythrocytes of Cyprinus carpio under the binary exposure of bifenthrin and chlorpyrifos by using single cell gel electrophoresis (SCGE). Limited efforts have been made to study the genotoxic effect for long duration period. Therefore, the present investigation was aimed to assess the genotoxicity of pesticide mixture to the freshwater carp, Cyprinus carpio at sub-lethal concentration exposure (33% LC50). At first 96-hr LC50 value of pesticide, the mixture was determined for Cyprinus carpio in a static system and then sub-lethal concentration was calculated and fish was exposed to this sub-lethal concentration of the mixture in glass aquaria for 70 days (five fortnights) at constant laboratory conditions. Peripheral blood erythrocytes were taken on a fortnightly basis for the time-dependent DNA damage assessment in-terms of percentage of damaged cells, genetic damage index and a cumulative tail length of comets. Concentration-dependent increase in the percentage of DNA damaged cells were observed up to a 4th fortnight, followed by a slight decrease in the 5th fortnight. Similarly, statistically significant time-dependent DNA damage was observed in terms of percentage of damaged cells, genetic damage index and a cumulative tail length of comets in treated fish (at 33% of LC50) as compared to control groups. The results supported the use of SCGE for evaluating the toxicity of pollutants which may be used as part of environmental monitoring programs.


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