Scientific Officer, Sri Lanka Atomic Energy Board
The aim of this proposal is to assess background radiation in relation to both cytogenetic dosimetry
This study aims to assess background frequency of micronuclei formation and chromosomal aberrations in persons living in Ragama and Norochcholai using biodosimetry. Biodosimetry is a technique used to assess exposure to radiation which is superior to physical dosimetry. It gives a clearer picture of individual variation of susceptibility to radiation as well as the ability to perform biodosimetry for human risk assessment. This study is important to establish baseline levels of micronuclei formation and chromosomal aberrations especially as a nuclear power plant is to be commissioned in nearby India recently. Biodosimetry has been used to detect external and internal exposure to ionising radiations in scenarios of accidental and occupational exposure wherever nuclear power plants are present (IAEA, 2012). This study proposes to study samples of 92 subjects (in the age range of 20-60 years), comprising 46 from Norochcholai (area closest to the power plant) and 46 (matched for gender, age and life style) from Ragama (as a control). Background radiation levels in the atmosphere will be recorded by a Geiger counter with the assistance of the Atomic Energy Authority and trace elements in soil and water samples will be assessed using Gamma spectrometry and X- Ray Flouresence technique, respectively. Venous blood samples will be obtained and processed at the Biodosimetry laboratory at the Faculty of Medicine, University of Kelaniya. 1000 binucleate cells will be assessed and the mean spontaneous micronuclei formation will be estimated. 200 metaphases will be analyzed for chromosomal aberrations. The results of this study can be used as baseline data to monitor exposure to radiation in the selected population in the future.
Abstract: The increasing risk of acute large-scale radiological/nuclear exposures of population underlines the necessity of developing new, rapid and high throughput biodosimetric tools for estimation of received dose and initial triage. We aimed to compare the induction and persistence of different radiation exposure biomarkers in human peripheral blood in vivo. Blood samples of patients with indicated radiotherapy (RT) undergoing partial body irradiation (PBI) were obtained soon before the first treatment and then after 24 h, 48 h, and 5 weeks; i.e. after 1, 2, and 25 fractionated RT procedures. We collected circulating peripheral blood from ten patients with tumor of endometrium (1.8 Gy per fraction) and eight patients with tumor of head and neck (2.0-2.121 Gy per fraction). Incidence of dicentrics and micronuclei was monitored as well as determination of apoptosis and the transcription level of selected radiation-responsive genes. Since mitochondrial DNA (mtDNA) has been reported to be a potential indicator of radiation damage in vitro, we also assessed mtDNA content and deletions by novel multiplex quantitative PCR. Cytogenetic data confirmed linear dose-dependent increase in dicentrics (p < 0.01) and micronuclei (p < 0.001) in peripheral blood mononuclear cells after PBI. Significant up-regulations of five previously identified transcriptional biomarkers of radiation exposure (PHPT1, CCNG1, CDKN1A, GADD45, and SESN1) were also found (p < 0.01). No statistical change in mtDNA deletion levels was detected; however, our data indicate that the total mtDNA content decreased with increasing number of RT fractions. Interestingly, the number of micronuclei appears to correlate with late radiation toxicity (r2 = 0.9025) in endometrial patients suggesting the possibility of predicting the severity of RT-related toxicity by monitoring this parameter. Overall, these data represent, to our best knowledge, the first study providing a multiparametric comparison of radiation biomarkers in human blood in vivo, which have potential for improving biological dosimetry.
Pub.: 24 Feb '18, Pinned: 11 Apr '18
Abstract: For mutagenicity testing, primary lymphocytes or mammalian cell lines are employed. However, the true target for carcinogenic action of mutagenic chemicals may be stem cells. Since hematopoietic cancers induced by chemical agents originate at the hematopoietic stem cell (HSC) stage and since one of the side effects of chemotherapeutic cancer treatment is the induction of secondary tumors, often leukemias, HSC may be a suitable cell system. We compared the sensitivity of HSC with the genotoxicity testing cell line TK6 for chromosomal mutations. HSC were less sensitive than TK6 cells for the genotoxic effects of the model genotoxins and chemotherapeutic agents doxorubicin, vinblastine, methyl methanesulfonate (MMS) and equally sensitive for mitomycin C (MMC). However, loss of viability after mitomycin C treatment was higher in HSC than in TK6 cells. Among the factors that may influence sensitivity for genomic damage, the generation or response to reactive oxygen species (ROS) and the effectiveness of DNA damage response can be discussed. Here we show that HSC can be used in a standard micronucleus test protocol for chromosomal mutations and that their sensitivity was not higher than that of a classical testing cell line.
Pub.: 22 Feb '18, Pinned: 11 Apr '18
Abstract: The aim of the present study was to monitor genotoxic and cytotoxic effect of X-ray on exfoliated buccal mucosa cells and investigate the association between the effects and the accumulated absorbed doses of oral mucosa. 98 participants' buccal mucosa cells were collected before and 10 days after different series of dental radiographs performed. Cytological preparations were successively dyed with the methods of Feulgen and fast-green, and analyzed under a light microscope. Micronuclei (MN)and other cells were scored. Accumulated absorbed dose of buccal mucosa was estimated with the method of anthropomorphic phantom and dosimeter chips. The dose rang was 0.18-3.54 mGy. A significant difference in the rate of MN cell was found before and after X-ray examinations (P = 0.008) as well as in the rates of Pyknotic (p < 0.001) and Karyolytic cell (p = 0.0021). When only the patients whose mucosa absorbed dose is lower than 1 mGy was analyzed, significant differences were not found except for Karyolytic cells (p = 0.0313). There was a correlation between the accumulated does and the change rate (ρ = 0.25, p = 0.0118). The frequency of micronuclei cells in buccal mucosa may be increased when a series of dental radiographs including a CBCT examination was performed.
Pub.: 08 Feb '18, Pinned: 11 Apr '18
Abstract: Lagging chromosomes that arise after chromosome mis-segregation during cell division can be encapsulated within small structures known as micronuclei. A link between whole-chromosome mis-segregation and chromothripsis has been demonstrated via micronuclear chromosome pulverization. Here, we describe methods to efficiently generate micronuclei and examine downstream cell fates, specifically with regard to DNA damage and chromosome pulverization.
Pub.: 23 Mar '18, Pinned: 11 Apr '18