Genotoxicity of Gamma Radiation Against Lymphocytes of Radiation Workers: The Cytokinesis-Block Micronucleus Assay.

<b>Background and Objective:</b> Gamma irradiation induces genotoxicity, characterized by the formation of extra-nuclear bodies and left behind during the anaphase stage of cell division, often referred to as a micronucleus (MN). The present work aims to monitor exposure to ionizing radiation as a genotoxic agent in the lymphocytes of workers at radiation energy centers. <b>Materials and Methods:</b> The lymphocyte cytokinesis block micronucleus assay used and analyzed the correlation between the Nuclear Division Index (NDI), age, blood type and the number of micronuclei (MN). Blood samples were collected from 20 volunteers in heparin tubes, exposed to 2 Gy gamma rays and cultured <i>in vitro</i>. <b>Results:</b> A significant difference in the number of micronuclei between blood group A and blood groups A, B and AB. The Nuclear Division Index (NDI) value for lymphocytes of radiation energy center workers after gamma radiation was significant (1.74±0.1) but still within the normal range. Neither MN frequency nor NDI values correlated with age, but MN frequency showed a correlation with blood type. <b>Conclusion:</b> The gamma irradiation did not induce a cytostatic effect but proved genotoxic to the lymphocytes of radiation energy center workers. Notably, blood type A demonstrated higher sensitivity to gamma radiation.

Study of Immune Response and Malondialdehyde Levels in Irradiated Rats Supplemented with Curcuma xanthorriza Roxb Extract.

OBJECTIVE: The purpose of this study was to assess the immune response and malondialdehyde levels in irradiated rats supplemented with Curcuma xanthorriza Roxb extract as a candidate for mitigating radiation exposure. METHODS: Twenty-four male Wistar rats were grouped into eight treatment groups, then Curcuma xanthorrhiza Roxb extract was administered orally and irradiated at 6 Gy. Measurement of rats IL-6 and INF-γ was performed using a sandwich ELISA Kit, while the MDA concentration was quantified according to the method of Wills (1971). The statistical test is determined by one way ANOVA test. P-value <0.05 was considered statistically significant. RESULT: The concentration of IL-6 in all groups showed no statistically significant difference (P=0.18). There was an increase in the concentration of IL-6 in the group of rats irradiated with 6 Gy for 7 days and 14 days. Meanwhile, the INF-γ concentration also showed no significant results in all treatment groups (P=0.28). The average of MDA concentration showed a significant difference in the liver and spleen of irradiated rats at 6 Gy for 14 days compared to the control (0.044 nmol/mg vs 0.008 nmol/mg, P=0.03 and 0.032 nmol/mg vs 0.014 nmol/mg, P=0.05, respectively). CONCLUSION: The administration of Curcuma xanthorriza Xorb extract was able to reduce MDA concentrations in the liver and spleen although not statistically significant. In addition, exposure to ionizing radiation at a dose of 6 Gy significantly increased lipid peroxidation in the liver and spleen by 5.5 times and 2.3 times, respectively.

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

OBJECTIVE: 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. METHODS: 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. RESULT: 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. CONCLUSION: The single-nucleotide polymorphism of hOGG1 exon 7 (rs1052133) demonstrated no association with the extent of DNA damage in radiation-exposed workers.

Superoxide dismutase and glutathione peroxidase activities in a population exposed to high indoor radon concentration: a preliminary report.

The objective of this study was to assess the effect of radon exposure on the modulation of endogenous antioxidants in a population chronically exposed to high levels of radon indoors. To do so, we measured the activities of superoxide dismutase (SOD) and glutathione peroxidase (GPX) in peripheral blood mononuclear cells (PBMCs) of people living in an area with high indoor radon concentration (Tande-Tande sub-village, Indonesia). The activities of SOD and GPX in Tande-Tande inhabitants were compared with those in subjects living in the Topoyo village (Indonesia), an area with low indoor radon levels. The activities of SOD and GPX in Tande-Tande sub-village inhabitants did not differ from those in people living in the Topoyo village (0.37 ± 0.021 versus 0.28 ± 0.018 U/mg protein and 8.46 ± 1.48 versus 8.34 ± 1.65 U/mg protein, p > .05). For both populations, there was a significant positive correlation between SOD and GPX activities (p < .001). No significant effects of gender, age, smoking habit, and body mass index on SOD and GPX activities were found for both groups. Although no significant modulation of SOD and GPX activities in PBMCs was detected, further studies should expand the sample size and also assess antioxidant levels in the serum. This study provides a first picture of endogenous antioxidant systems in Tande-Tande sub-village inhabitants, but a more comprehensive analysis, including the measurement of catalase (CAT) activity, might provide additional insight into the effects of chronic exposure to high indoor radon concentrations.

Development of Dose-Response Calibration Curve for Dicentric Chromosome Induced by X-Rays.

Chromosome aberration is a biomarker that has been used as a standard tool in biological dosimetry (biodosimetry) of individuals after exposure to ionizing radiation. It is based mainly on the induction of dicentric chromosomes - one of the radiation-induced biological effects, in order to correlate them with radiation dose. In this study, a dose calibration curve for X-rays was generated by using the dicentric assay and by fitting the data to both Chromosomal Aberration Calculation Software and Dose Estimate programs to compare the output of each method. Peripheral blood samples from four nonsmoker healthy donors were irradiated with various doses ranging from 0 to 4 Gy with 250 kV or 122 keV X-rays at a dose rate of 0.17 Gy/min. The irradiated blood was cultured, harvested, and analyzed according to the standard procedure as described by the International Atomic Energy Agency with slight modifications. The dose-response calibration data for dicentrics were fitted with the linear-quadratic model (Ydic = 0.03987D2 + 0.00651D). The dose-response calibration curve obtained in this research was comparable to other estimations with similar radiation quality and dose rates. The results in this research convinced us in sustaining a biodosimetry using a dose-response calibration curve in our laboratory.

Study of γ-H2AX as DNA double strand break biomarker in resident living in high natural radiation area of Mamuju, West Sulawesi.

High expression of phospho histone γ-H2AX, a sensitive marker of double stranded DNA damage, is believed to be an indication of defective DNA repair pathway or genomic instability that may cause mutations and ultimately cancer. DNA damage can be caused by ionizing radiation exposure. Beside in medical treatment/diagnosis or industry, ionizing radiation exposure can also be found in naturally in regions of high natural back ground radiation. In this study we collect the blood from 45 volunteers living in Mamuju, a region with highest natural radiation in Indonesia (dose of ∼7 mSv/year). Subjects were grouped as high natural background area (HNBA) (n = 37) and control area (n = 8). The expression γ-H2AX foci were evaluated by one of researcher fluorescence microscope examination. Our results show that the average foci numbers per cell were in the normal range. While not statistical different, the average of γ-H2AX foci in exposed area higher in the exposed compared to the control area, 0.31 versus 0.13 (p > 0.05), respectively. Moreover, there was also no statistical difference of average γ-H2AX foci between man and woman, old and young people in exposed and control area (p > 0.05). In this preliminary study we find that γ-H2AX foci (and thus DNA double strand break) frequency in residents living in the HNBA of Mamuju, West Sulawesi, show a trend towards higher (albeit not significant) average values relative to the control area. More research is needed to further scrutinize these observations.

Establishment of a Dose-response Curve for X-ray-Induced Micronuclei in Human Lymphocytes.

The cytokinesis-block micronucleus assay in peripheral blood lymphocytes is an established technique for biodosimetry. The aim of this project was to generate a X-ray induced micronuclei (MN) curve for peripheral blood lymphocytes taken from five healthy donors. The blood samples were irradiated with X-rays of 122 KeV at a dose rate of 0.652 Gy/min to doses of 0.5, 1, 2, 3, and 4 Gy. The blood samples were then cultured for 72 h at 37°C and processed following the International Atomic Energy Agency standard procedure with slight modifications. The result showed that the yields of MN frequencies were increased with the increase of radiation dose. Reconstruction of the relationship of MN with dose was fitted to a linear-quadratic model using Chromosome Aberration Calculation Software version 2.0. Due to their advantages, mainly, the dependence on radiation dose and dose rate, despite their limitation, these curves will be useful as alternative method for in vitro dose reconstruction and can support the preparedness for public or occupational radiation overexposure and protection. The results reported here also give us confidence to apply the obtained calibration curve of MN for future biological dosimetry requirements in Indonesia.