Normal Level Natural Radiation Areas Research Articles

Non-linear dose response of DNA double strand breaks in response to chronic low dose radiation in individuals from high level natural radiation areas of Kerala coast

BackgroundThe human population living in high level natural radiation areas (HLNRAs) of Kerala coast provide unique opportunities to study the biological effects of low dose and low dose rate ionizing radiation below 100 mGy. The level of radiation in this area varies from < 1.0 to 45 mGy/year. The areas with ≤ 1.50 mGy/year are considered as normal level natural radiation areas (NLNRA) and > 1.50 mGy/year, as high level natural radiation areas (HLNRA). The present study evaluated dose response relationship between DNA double strand breaks (DSBs) and background radiation dose in individuals residing in Kerala coast. Venous blood samples were collected from 200 individuals belonging to NLNRA (n = 50) and four dose groups of HLNRA; 1.51-5.0 mGy/year (n = 50), 5.01-10.0 mGy/year (n = 30), 10.01-15.0 mGy/year (n = 33), > 15.0 mGy/year (n = 37) with written informed consent. The mean dose of NLNRA and four HLNRA dose groups studied are 1.21 ± 0.21 (range: 0.57–1.49), 3.02 ± 0.95 (range: 1.57–4.93), 7.43 ± 1.48 (range: 5.01–9.75), 12.22 ± 1.47 (range: 10.21–14.99), 21.64 ± 6.28 (range: 15.26–39.88) mGy/year, respectively. DNA DSBs were quantified using γH2AX as a marker, where foci were counted per cell using fluorescence microscopy.ResultsOur results revealed that the frequency of γH2AX foci per cell was 0.090 ± 0.051 and 0.096 ± 0.051, respectively in NLNRA and HLNRA individuals, which were not significantly different (t198 = 0.33; P = 0.739). The frequency of γH2AX foci was observed to be 0.090 ± 0.051, 0.096 ± 0.051, 0.076 ± 0.036, 0.087 ± 0.042, 0.108 ± 0.046 per cell, respectively in different dose groups of ≤ 1.50, 1.51-5.0, 5.01-10.0, 10.01-15.0, > 15.0mGy/year (ANOVA, F4,195 = 2.18, P = 0.072) and suggested non-linearity in dose response. The frequency of γH2AX foci was observed to be 0.098 ± 0.042, 0.078 ± 0.037, 0.084 ± 0.042, 0.099 ± 0.058, 0.097 ± 0.06 and 0.114 ± 0.033 per cell in the age groups of ≤ 29, 30–34, 35–39, 40–44, 45–49 and ≥ 50 years, respectively (ANOVA, F5,194 = 2.17, P = 0.059), which suggested marginal influence of age on the baseline of DSBs. Personal habits such as smoking (No v/s Yes: 0.092 ± 0.047 v/s 0.093 ± 0.048, t198 = 0.13; P = 0.895) and drinking alcohol (No v/s Yes: 0.096 ± 0.052 v/s 0.091 ± 0.045, t198 = 0.62; P = 0.538) did not show any influence on DSBs in the population.ConclusionThe present study did not show any increase in DSBs in different dose groups of HLNRA compared to NLNRA, however, it suggested a non-linear dose response between DNA DSBs and chronic low dose radiation.

Evaluation of natural chronic low dose radiation exposure on telomere length and transcriptional response of shelterin complex in individuals residing in Kerala coast, India

The high level natural radiation areas (HLNRA) of Kerala coast provide unique opportunity to study the biological effect of chronic low dose ionizing radiation (LDIR) on human population below 100 mSv. The radiation level in this area varies from < 1.0–45 mGy /year due to patchy distribution of monazite in the sand, which contains 232Th (8–10%), 238U (0.3%), and their decay products. Telomere length attrition has been correlated to DNA damage due to genotoxic agents. The objective of the present study is to evaluate the effect of natural chronic LDIR exposure on telomere length and transcriptional response of telomere specific and DNA damage repair genes in peripheral blood mononuclear cells (PBMCs) of individuals from normal level natural radiation areas (NLNRA) and HLNRA of Kerala coast, southwest India. Blood samples were collected from 71 random male donors (24–80 years) from NLNRA (≤1.50 mGy/year; N = 19) and two HLNRA dose groups [1.51–10 mGy/year (N = 17); > 10 mGy/year, (N = 35)]. Genomic DNA was isolated from PBMCs and relative telomere length (RTL) was determined using real time q-PCR. Radio-adaptive response (RAR) study was carried out in PBMCs of 40 random males from NLNRA (N = 20) and HLNRA (>10 mGy/year; N = 20), where PBMCs were given a challenged dose of 2.0 Gy gamma radiation at 4 h. Transcriptional profile of telomere specific (TRF1, TRF2, POT1, TIN2, TPP1, RAP1), DNA damage response (RAD17, ATM, CHEK1) and base excision repair pathway (BER) (OGG1, XRCC1, NTH1, NEIL1, MUTYH, MBD4) genes were analysed at basal level and after a challenge dose of 2.0 Gy at 4 h. Our results did not show any significant effect of chronic LDR on RTL among the individuals from NLNRA and two HLNRA groups (p = 0.195). However, influence of age on RTL was clearly evident among NLNRA and HLNRA individuals. At basal level, TRF1, TRF2, TIN2, MBD4, NEIL1 and RAD17 showed significant up-regulation, whereas XRCC1 was significantly down regulated in HLNRA individuals. After a challenge dose of 2.0 Gy, significant transcriptional up-regulation was observed at telomere specific (TRF2, POT1) and BER (MBD4, NEIL1) genes in HLNRA individuals as compared to NLNRA suggesting their role in RAR. In conclusion, elevated level of natural chronic LDR exposure did not have any adverse effect on telomere length in Kerala coast. Significant transcriptional response at TRF2, MBD4 and NEIL1 at basal level and with a challenge dose of 2.0 Gy suggested their active involvement in efficient repair and telomere maintenance in individuals from HLNRA of Kerala coast.

Evaluation of Spontaneous DNA Damage Using the Alkaline Comet Assay in Lymphocyte Cells of Humans Living in the High Level Natural Radiation Area of Mamuju, Indonesia

To evaluate the biological impacts of high background radiation exposures that are represented by spontaneous deoxyribonucleic acid (DNA) damage, an evaluation on lymphocyte cells from residents of Mamuju, West Sulawesi, Indonesia was tested. The mean annual dose received by individuals in this area is about 10.40 mSv. Of the 177 adult subjects studied, 102 were from high-level natural radiation areas of Mamuju and 75 subjects were from a nearby normal-level natural radiation area. Both areas are similar in living situations. DNA strand breaks and other parameters of study and control group were determined using a standardized comet assay. Our results showed that chronic low-level natural radiation had resulted in significantly higher (p&lt;0.001) DNA damage based on the three parameters of the assay (tail length, tail DNA, and tail moment) compared to those of control. There was a positive correlation between the level of DNA damage and age, where people aged 40 years and older had a higher level of DNA damage than those under 40 year. The level of DNA damage was also found to be higher in females compared to that of males. It was concluded that chronic exposure to natural radiation in Mamuju had induced spontaneous DNA damage in human cells after long-term exposure which was dependent on age and sex.

Evaluation of risk due to chronic low dose ionizing radiation exposure on the birth prevalence of congenital heart diseases (CHD) among the newborns from high-level natural radiation areas of Kerala coast, India

BackgroundThe human population residing in monazite bearing Kerala coast are exposed to chronic low dose and low dose rate external gamma radiation due to Th232 deposits in its beach sand. The radiation level in this area varies from < 1.0 to 45.0 mGy/year. This area serves as an ideal source for conducting large-scale epidemiological studies for assessing risk of low dose and low dose rate radiation exposure on human population. The areas with a dose level of ≤1.50 mGy/year are considered as normal level natural radiation areas (NLNRAs) and areas with > 1.50 mGy/year, as high level natural radiation areas (HLNRAs). HLNRAs were further stratified into three dose groups of 1.51-3.0 mGy/year, 3.01-6.00 mGy/year and > 6.0 mGy/year. The present study evaluates the effects of chronic low dose radiation (LDR) exposure on the birth prevalence of Congenital Heart Diseases (CHD) among the live newborns monitored in hospital based prospective study from NLNRAs and HLNRAs of Kerala coast, India.MethodologyConsecutive newborns were monitored from two hospital units located in the study area for congenital malformations. Referred CHD cases among the newborns screened were confirmed by conducting investigations such as pulse oximetry, chest X-ray, electrocardiogram and echocardiogram etc.ResultsAmong the newborns screened, 289 CHDs were identified with a frequency of 1.49‰ among 193,634 livebirths, which constituted 6.03% of overall malformations and 16.29% of major malformations. Multiple logistic regression analysis suggested that the risk of CHD among the newborns of mothers from HLNRAs with a dose group of 1.51-3.0 mGy/year was significantly lower as compared to NLNRA (OR = 0.72, 95% CI: 0.57-0.92), whereas it was similar in HLNRA dose groups of 3.01-6.00 mGy/year (OR = 0.55, 95% CI: 0.31-1.00) and ≥ 6.0 mGy/year (OR = 0.96, 95% CI: 0.50-1.85). The frequency of CHDs did not show any radiation dose related increasing trend. However, a significant (P = 0.005) reduction was observed in the birth prevalence of CHDs among the newborns from HLNRA (1.28‰) as compared to NLNRA (1.79‰).ConclusionChronic LDR exposure did not show any increased risk on the birth prevalence of CHDs from high-level natural radiation areas of Kerala coast, India. No linear increasing trend was observed with respect to different background dose groups. The frequency of CHD was observed to be 1.49 per 1000 livebirths, which was similar to the frequency of severe CHD rate reported elsewhere in India and was much less than the reported frequency of 9 per thousand.

Evaluation of the influence of chronic low-dose radiation on DNA repair gene polymorphisms [XRCC1, XRCC3, PRKDC (XRCC7), LIG1, NEIL1] in individuals from normal and high level natural radiation areas of Kerala Coast

Background: Single Nucleotide Polymorphisms (SNPs) at DNA repair genes are considered as potential biomarkers of radio-sensitivity. The coastal belt of Kerala in south west India has a patchy distribution of monazite in its beach sand that contains Th-232 and its decay products. Thus, radiation levels in this area vary from <1.0mGy to 45.0mGy/year. The areas with external gamma radiation dose >1.5mGy/year are considered as High-Level Natural Radiation Areas (HLNRA) and ≤ 1.5mGy/year are Normal Level Natural Radiation Area (NLNRA).Objective: In the present study, an attempt was made to evaluate the influence of chronic low dose radiation exposure on DNA repair gene polymorphisms in NLNRA and HLNRA population of Kerala coast.Materials and methods: Genomic DNA was isolated from venous blood samples of 246 random, healthy individuals (NLNRA, N = 104; HLNRA, N = 142) and genotyping of five SNPs such as X-ray repair cross complementing 1(XRCC1 Arg399Gln), X-ray repair cross complementing 3 (XRCC3 Thr241Met], Protein kinase, DNA-activated, catalytic subunit (PRKDC) (X-ray repair cross-complementing group 7, XRCC7 G/T), nei like DNA glycosylase 1 (NEIL1 G/T) and DNA ligase 1 (LIG1 A/C) was carried out using PCR based restriction fragment length polymorphism (PCR-RFLP) followed by silver staining.Results: Our results showed no significant difference in genotype frequencies in HLNRA vs NLNRA at three of the five SNPs studied i.e. XRCC1 Arg399Gln (χ2(2) = 5.85, p = .054), XRCC3 Thr241Met (χ2(1) = 0.71, p = .339), PRKDC (XRCC7 G/T) (χ2(2) = 3.72, p = .156), whereas significant difference was observed at NEIL1 G/T (χ2(2) =8.71, p = .013) and LIG1 A/C (χ2(2) = 7.66, p = .022). The odds of heterozygote to homozygote genotypes in HLNRA relative to NLNRA at XRCC1 Arg399Gln (OR = 1.96, 95% CI: 1.13–3.40), XRCC3 Thr241Met (OR = 0.73, 95% CI: 0.41–1.31), PRKDC (XRCC7 G/T), (OR = 0.81; 95% CI: 0.48–1.38), NEIL1 G/T (OR = 0.54; 95% CI: 0.31–0.96) and LIG1 A/C (OR = 1.62; 95% CI: 0.97–2.69) was also not significantly different in HLNRA vs NLNRA, except at XRCC1 and NEIL1.Conclusion: The genotype frequencies at three of these SNPs i.e. XRCC1 Arg399Gln, XRCC3 Thr241Met and PRKDC (XRCC7 G/T) were similar, whereas NEIL1 G/T and LIG1 A/C showed significant difference between HLNRA and NLNRA population. However, further research using more number of SNPs in a larger cohort is required in this study area.

Frequency of chromosome aberrations among adult male individuals from high and normal level natural radiation areas of Kerala in the southwest coast of India

Chromosome aberration analysis was carried out in peripheral blood lymphocytes of adult male individuals from normal level natural radiation areas (NLNRA, ≤1.5 mGy/year, N = 27) and high level natural radiation areas (HLNRA, >1.5mGy/year, N = 70) of Kerala coast in southwest India. The mean age of individuals from NLNRA and HLNRA was 40.9 ± 9.4 and 43.7 ± 12.4 years, respectively, with an overall mean of 42.9 ± 11.6 (range: 18–80). Whole-blood cultures were set up and about 260 metaphases were scored per individual. The frequency of chromosome aberrations was calculated per 1000 cells. The overall basal frequency of unstable (dicentrics and rings), stable (translocations and inversions) and other (fragments and breaks) aberrations was 1.54 ± 0.25, 4.1 ± 0.40 and 6.66 ± 0.51, respectively. Individuals of NLNRA and HLNRA had statistically similar frequency of unstable (2.11 ± 0.64 v/s 1.39 ± 0.26; RR = 0.66; 95% CI: 0.33-1.33), stable (4.60 ± 0.94 v/s 3.97 ± 0.44; RR = 0.86; 95% CI: 0.55-1.36) and other (7.85 ± 1.23 v/s 6.36 ± 0.56; RR = 0.81; 95% CI: 0.57-1.15) chromosome aberrations. Frequencies of unstable, stable and other chromosome aberrations did not show any dose response after stratification of HLNRA samples into three dose groups (1.51–5.0 mGy/year, 5.01–10 mGy/year and >10.0 mGy/year). Smokers showed an increase in other chromosome aberrations (P < 0.001), but smoking was not associated with unstable and stable aberrations. Alcohol consumption and tobacco chewing had no significant association with any type of chromosome aberrations. In conclusion, chronic low dose radiation prevailing in Kerala coast did not show any significant effect on the basal frequency of chromosome aberrations among the adult population.

Efficient repair of DNA double strand breaks in individuals from high level natural radiation areas of Kerala coast, south-west India

High level natural radiation areas (HLNRA) of Kerala coastal strip (55km long and 0.5km wide) in southwest India exhibit wide variations in the level of background dose (< 1.0–45.0mGy/year) due to thorium deposits in the beach sand. The areas with ≤1.5mGy/year are considered as normal level natural radiation area (NLNRA), whereas areas with >1.5mGy/year are HLNRA. Individuals belonging to HLNRA were stratified into two groups, Low dose group (LDG: 1.51–5.0mGy/year) and high dose group (HDG: >5.0mGy/year). The mean annual dose received by the individuals from NLNRA, LDG and HDG was 1.3±0.1, 2.7±0.9 and 9.4±2.3mGy/year, respectively. Induction and repair of DNA double strand breaks (DSBs) in terms of gamma-H2AX positive cells were analysed in peripheral blood mononuclear cells (PBMCs) using flow cytometry. Induction of DSBs was studied at low (0.25Gy) and high challenge doses (1.0 and 2.0Gy) of gamma radiation in 78 individuals {NLNRA, N=23; HLNRA (LDG, N=21 and HDG, N=34)}. Repair kinetics of DSBs were evaluated in PBMCs of 30 individuals belonging to NLNRA (N=8), LDG (N=7) and HDG (N=15) at low (0.25Gy) and high doses (2.0Gy) of gamma radiation. Transcription profile of DNA damage response (DDR) and DSB repair genes involved in non-homologous end joining (NHEJ) and homologous recombination repair (HRR) pathways was analysed after a challenge dose of 2.0Gy in PBMCs of NLNRA (N=10) and HDG, HLNRA (N=10) group. Our results revealed significantly lower induction and efficient repair of DSBs in HLNRA groups as compared to NLNRA. Transcription profile of DCLRE1C, XRCC4, NBS1 and CDK2 showed significant up-regulation (p≤0.05) in HDG at a challenge dose of 2.0Gy indicating active involvement of DDR and DSB repair pathways. In conclusion, lower induction and efficient repair of DNA DSBs in HLNRA groups is suggestive of an in vivo radio-adaptive response due to priming effect of chronic low dose radiation prevailing in this area.

Radio-adaptive response in peripheral blood lymphocytes of individuals residing in high-level natural radiation areas of Kerala in the southwest coast of India.

The present study investigates whether the chronic low-dose radiation exposure induces an in vivo radio-adaptive response in individuals from high-level natural radiation areas (HLNRA) of the Kerala coast. Peripheral blood samples from 54 adult male individuals aged between 26 and 65 years were collected for the study with written informed consent. Each of the whole blood sample was divided into three, one was sham irradiated, second and third was exposed to challenging doses of 1.0 and 2.0 Gy gamma radiation, respectively. Cytokinesis-block micronucleus (CBMN) assay was employed to study the radio-adaptive response. Seventeen individuals were from normal-level natural radiation area (NLNRA ≤1.5 mGy/year) and 37 from HLNRA (> 1.5 mGy/year). Based on the annual dose received, individuals from HLNRA were further classified into low-dose group (LDG, 1.51-5.0 mGy/year, N = 19) and high-dose group (HDG >5.0 mGy/year, N = 18). Basal frequency of micronucleus (MN) was comparable across the three dose groups (NLNRA, LDG and HDG, P = 0.64). Age of the individuals showed a significant effect on the frequency of MN after challenging dose exposures. The mean frequency of MN was significantly lower in elder (>40 years) individuals from HDG of HLNRA as compared to the young (≤40 years) individuals after 1.0 Gy (P < 0.001) and 2.0 Gy (P = 0.002) of challenging doses. However, young and elder individuals within NLNRA and LDG of HLNRA showed similar frequency of MN after the challenging dose exposures. Thus, increased level of chronic low-dose radiation (>5.0 mGy/year) seems to act as a priming dose resulting in the induction of an in vivo radio-adaptive response in elder individuals of the Kerala coast.

Lack of increased DNA double-strand breaks in peripheral blood mononuclear cells of individuals from high level natural radiation areas of Kerala coast in India

The high level natural radiation area (HLNRA) of Kerala is a 55km long and 0.5km wide strip in south west coast of India. The level of background radiation in this area varies from <1.0mGy/year to 45.0mGy/year. It offers unique opportunity to study the effect of chronic low dose/low dose-rate radiation directly on human population. Spontaneous level of DNA double strand breaks (DSBs) was quantified in peripheral blood mononuclear cells of 91 random individuals from HLNRA (N=61, mean age: 36.1±7.43years) and normal level natural radiation area (NLNRA) (N=30, mean age: 35.5±6.35years) using gamma-H2AX as a marker. The mean annual dose received by NLNRA and HLNRA individuals was 1.28±0.086mGy/year and 8.28±4.96mGy/year, respectively. The spontaneous frequency of DSBs in terms of gamma-H2AX foci among NLNRA and HLNRA individuals were 0.095±0.009 and 0.084±0.004 per cell (P=0.22). The individuals from HLNRA were further classified as low dose group (LDG, 1.51–5.0mGy/year, mean dose: 2.63±0.76mGy/year) and high dose group (HDG, >5.0mGy/year, mean dose: 11.04±3.57mGy/year). The spontaneous frequency of gamma-H2AX foci per cell in NLNRA, LDG and HDG was observed to be 0.095±0.009, 0.096±0.008 and 0.078±0.004 respectively. Individuals belonging to HDG of HLNRA showed marginally lower frequency of DSBs as compared to NLNRA and LDG of HLNRA. This could be suggestive of either lower induction or better repair of DSBs in individuals from HDG of HLNRA. The present study indicated that 5.0mGy/year could be a possible threshold dose for DSB induction at chronic low-dose radiation exposure in vivo. However, further studies on DNA damage induction and repair kinetics are required to draw firm conclusions.

Effect of chronic low dose natural radiation in human peripheral blood mononuclear cells: Evaluation of DNA damage and repair using the alkaline comet assay

This study investigates whether peripheral blood mononuclear cells (PBMCs) from inhabitants of Kerala in southwest India, exposed to chronic low dose natural radiation in vivo (>1mSvyear−1), respond with a radioadaptive response to a challenging dose of gamma radiation. Toward this goal, PBMCs isolated from 77 subjects from high-level natural radiation areas (HLNRA) and 37 subjects from a nearby normal level natural radiation area (NLNRA) were challenged with 2Gy and 4Gy gamma radiation. Subjects from HLNRA were classified based on the mean annual effective dose received, into low dose group (LDG) and high dose group (HDG) with mean annual effective doses of 2.69mSv (N=43, range 1.07mSvyear−1 to 5.55mSvyear−1) and 9.62mSv (N=34, range 6.07mSvyear−1 to17.41mSvyear−1), respectively. DNA strand breaks and repair kinetics (at 7min, 15min and 30min after 4Gy) were evaluated using the alkaline single cell gel electrophoresis (comet) assay. Initial levels of DNA strand breaks observed after either a 2Gy or a 4Gy challenging dose were significantly lower in subjects of the HDG from HLNRA compared to subjects of NLNRA (2Gy, P=0.01; 4Gy, P=0.02) and LDG (2Gy P=0.01; 4Gy, P=0.05). Subjects of HDG from HLNRA showed enhanced rejoining of DNA strand breaks (HDG/NLNRA, P=0.06) during the early stage of repair (within 7min). However at later times a similar rate of rejoining of strand breaks was observed across the groups (HDG, LDG and NLNRA). Preliminary results from our study suggest in vivo chronic low-level natural radiation provides an initial exposure that allows an adaptation to a subsequent higher radiation exposure, perhaps through improving DNA repair via an unknown mechanism. Therefore, further investigations would be necessary in this population to understand the biological and health effects of chronic low-level natural radiation exposures.

The effect of high level natural ionizing radiation on expression of PSA, CA19-9 and CEA tumor markers in blood serum of inhabitants of Ramsar, Iran

Since several high level natural radiation areas (HLNRAs) exist on our planet, considerable attention has been drawn to health issues that may develop as the result of visiting or living in such places. City of Ramsar in Iran is an HNLRA, and is a tourist attraction mainly due to its hot spas. However, the growing awareness over its natural radiation sources has prompted widespread scientific investigation at national level. In this study, using an ELISA method, the level of expression of three tumor markers known as carcinoembryonic antigen (CEA), prostate-specific antigen (PSA) and carcino antigen 19-9 (CA19-9) in blood serum of 40 local men of Ramsar (subject group) was investigated and compared to 40 men from the city of Noshahr (control group). Noshahr was previously identified as a normal level natural radiation area (NLNRA) that is some 85 km far from Ramsar. According to statistical analysis, there was a significant difference in the levels of PSA and CA19-9 markers between the two groups (p < 0.001) with those of Ramsar being considerably higher. CEA level did not show any difference. Although some of the volunteers tested positive to the markers, they were in good health as confirmed by the physician. Moreover, the high number of positive markers in Noshahr was considerable. Therefore, future study is needed to further validate this result and to determine the level of positivity to tumor markers in both cities.

Effects of chronic low level radiation in the population residing in the high level natural radiation area in Kerala, India: Employing heritable DNA mutation studies

To study the effect of chronic low level radiation, 4040 meiosis were screened at eight microsatellite and five minisatellite (2485 and 1555 meiosis respectively) marker loci in people residing in high and normal level natural radiation areas of Kerala. Variants in the repeat length of allele were considered as mutants. Mutation rates (expressed as the number of mutations observed in the total number of meiosis) were 6.4×10−3 (16/2485) and 2.6×10−3 (4/1555) at microsatellite and minisatellite respectively. The germline microsatellite mutation frequency of father was 1.78 times higher at 7.52×10−3 (8/1064) compared to 4.22×10−3 (6/1421) of mother (P=0.292, Fisher's Exact two-sided test). The paternal and maternal mutation rates at minisatellite loci were more or less similar at 2.78×10−3 (2/719) and 2.39×10−3 (2/836), respectively (P=1.0, Fisher's Exact two-sided test). Higher but statistically non-significant microsatellite mutation frequency was observed in HLNRA compared to NLNRA (7.25×10−3 vs 3.64×10−3; P=0.547). The apparent increase in the mutation rate of microsatellite loci with the increase in radiation dose was also not statistically significant. All the four minisatellite mutation observed were from HLNRA (1198 meiosis) and no mutation was observed among 357 meiosis screened from NLNRA families. All the markers used in the present study were in the non-coding region and hence mutations in these regions may not cause adverse health effects, but the study is important in understanding the effect of chronic low level radiation.

Cytogenetic studies on newborns from high and normal level natural radiation areas of Kerala in southwest coast of India

Purpose: To study, characterize and compare chromosome aberrations and karyotype anomalies among newborns from high (> 1.5 mGy/y) and normal (≤ 1.5 mGy/y) level natural radiation areas of monazite-sand bearing southwest coast of Kerala in India.Materials and methods: Cord blood samples from newborns were collected from selected Government hospitals in heparinized vials and cultures were set up employing standard microculture techniques, slides were prepared, coded and stained with giemsa. Well spread metaphases were analyzed for chromosome aberrations and karyotype anomalies.Results: A total of 1,267,788 metaphases from 27,295 newborns of mothers aged 17–45 years (17,298 from high and 9,997 from normal level radiation areas) were analyzed during 1986–2007. Frequencies of dicentrics in high and normal level radiation areas were 1.90 ± 0.14 and 2.01 ± 0.26 per 10,000 cells, respectively (Relative frequency [RF] = 0.94; 95% CI: 0.71–1.26). Karyotype anomalies had a frequency of 5.49‰ and 6.7‰, respectively (RF = 0.82; 95% CI: 0.60–1.12). No dose-related trend was observed in chromosome aberrations or karyotype anomalies.Conclusion: Frequencies of chromosomal aberration and karyotype anomalies between the newborns from the high level natural radiation area (HLNRA) and normal level natural radiation areas (NLNRA) were very similar.

No evidence of telomere length attrition in newborns from high level natural background radiation areas in Kerala coast, south west India

Purpose: The tandemly repeated hexamers (TTAGGG)n present at the telomeric ends protect the human genome from a variety of environmental exposures including ionizing radiation. In order to find out the effect of chronic low dose radiation exposure, we have determined telomere length among newborns from high level natural radiation areas (HLNRA) of the Kerala coast in South west India.Methodology: Umbilical cord blood samples were collected from 128 newborns from HLNRA and 43 newborns from normal level natural radiation areas (NLNRA) and genomic DNA was isolated using a salt precipitation method. The mean telomere length was determined using SYBR green-based real time quantitative Polymerase chain reaction (qPCR), where telomere gene specific (T) and single copy gene specific (S) primers were used. The average of telomere versus single copy gene (T/S) ratio was calculated which was proportional to telomere length of each individual.Results: The mean relative telomere length was found to be 1.03 ± 0.01 (95% CI, 0.99–1.05) and 1.10 ± 0.03 (95% CI, 1.04–1.17) in HLNRA and NLNRA newborns, respectively (P > 0.05). Based on the level of background radiation dose, samples were divided into four groups, i.e., NLNRA: < 1.50 mGy/year and three HNLRA groups: 1.51–3.00 mGy/year, 3.01–5.00 mGy/year, and > 5.00 mGy/year. The mean relative telomere length in these groups were found to be 1.10 ± 0.03 (95% CI, 1.03–1.17), 0.98 ± 0.01 (95% CI, 0.95–1.01), 1.05 ± 0.02 (95% CI, 1.01–1.10) and 1.07 ± 0.03 (95% CI, 1.04–1.10), respectively. No significant difference was observed between the mean telomere length of male and female newborns.Conclusions: The elevated level of natural chronic background radiation prevailing in Kerala coast did not show any significant effect on telomere length of newborns. To our knowledge, this is the first report on newborn telomere length from a high level natural radiation area.

Evaluation of Spontaneous DNA Damage in Lymphocytes of Healthy Adult Individuals from High-Level Natural Radiation Areas of Kerala in India

Inhabitants of the high-level natural radiation areas (>1 mSv year(-1)) of Kerala in southwest India were evaluated for basal damage (spontaneous DNA strand breaks and alkali-labile sites) by the alkaline comet assay and oxidative DNA damage (ENDO III- and hOGG1-sensitive sites) by the enzyme-modified comet assay. Of the 67 adult male subjects studied, 45 were from high-level natural radiation areas and 22 subjects were from a nearby normal-level natural radiation area (≤1 mSv year(-1)). Basal damage due to the age and residential area (normal-level natural radiation area/high-level natural radiation areas) of the donors showed significant interaction (P < 0.001) when all subjects were analyzed using a general linear model (GLM). In subgroup analysis, basal damage increased with age in subjects from the normal-level natural radiation area (P = 0.02), while a significant negative correlation (P = 0.002) was observed in subjects from high-level natural radiation areas. Further, basal damage in elderly subjects from high-level natural radiation areas was significantly (P < 0.001) lower compared to the subjects from the normal-level natural radiation area. Oxidative DNA damage was not influenced by age, smoking habit or residential area in the entire sample. However, in a subgroup analysis, hOGG1-sensitive sites showed a significant increase with age in subjects from high-level natural radiation areas (P = 0.005). ENDO III-sensitive sites increased with natural radiation exposure in subjects from high-level natural radiation areas (P = 0.02), but when stratified according to smoking, a significant increase was observed only in smokers (P = 0.01). To the best of our knowledge, this is the first study on basal and oxidative DNA damage in healthy adults of this population. However, our findings need more validation in a larger study population.

Radioadaptive responses induced in lymphocytes of the inhabitants in Ramsar, Iran

Ramsar, a city in northern Iran, has among the highest levels of natural radiation known to exist in an inhabited area. Twenty-two residents of high-level natural radiation areas and 33 residents from an adjacent normal-level natural radiation area participated in this study. In the first phase of the experiment, 15 healthy residents from high-level natural radiation areas and 30 healthy inhabitants of a nearby normal-level natural radiation area were studied. In the second phase, seven healthy residents with cumulative lifetime doses of up to 10 Sv were studied for assessing the induction of adaptive response in each study participant and obtaining complementary data. Cultured cells were given a challenge dose of either 2 Gy (first phase of the study) or 1.5 Gy (second phase of the study) of Co-60 gamma radiation. Overall data showed a significant radioadaptive response in the residents of high-level natural radiation areas. Results obtained in the second phase of the study showed that five out of seven inhabitants exhibited a reduction in induced chromosomal aberrations following exposure to a 1.5 Gy challenge dose of gamma radiation. As the cumulative dose increased from a few hundred mGy to 1 Gy, the magnitude of the induced adaptive response increased linearly.

Child cancer risk in high-background radiation areas

To study the effect of chronic low level radiation, 4040 meiosis were screened at eight microsatellite and five minisatellite (2485 and 1555 meiosis respectively) marker loci in people residing in high and normal level natural radiation areas of Kerala. Variants in the repeat length of allele were considered as mutants. Mutation rates (expressed as the number of mutations observed in the total number of meiosis) were 6.4 × 10−3 (16/2485) and 2.6 × 10−3 (4/1555) at microsatellite and minisatellite respectively. The germline microsatellite mutation frequency of father was 1.78 times higher at 7.52 × 10−3 (8/1064) compared to 4.22 × 10−3 (6/1421) of mother (P = 0.292, Fisher's Exact two-sided test). The paternal and maternal mutation rates at minisatellite loci were more or less similar at 2.78 × 10−3 (2/719) and 2.39 × 10−3 (2/836), respectively (P = 1.0, Fisher's Exact two-sided test). Higher but statistically non-significant microsatellite mutation frequency was observed in HLNRA compared to NLNRA (7.25 × 10−3 vs 3.64 × 10−3; P = 0.547). The apparent increase in the mutation rate of microsatellite loci with the increase in radiation dose was also not statistically significant. All the four minisatellite mutation observed were from HLNRA (1198 meiosis) and no mutation was observed among 357 meiosis screened from NLNRA families. All the markers used in the present study were in the non-coding region and hence mutations in these regions may not cause adverse health effects, but the study is important in understanding the effect of chronic low level radiation.

Genetic Monitoring of the Human Population from High-Level Natural Radiation Areas of Kerala on the Southwest Coast of India. II. Incidence of Numerical and Structural Chromosomal Aberrations in the Lymphocytes of Newborns

Cytogenetic studies using cord blood samples from newborns from high-level natural radiation areas of the Kerala coast in Southwest India have been in progress since 1986. A total of 963,940 metaphases from 10,230 newborns have been screened for various types of chromosomal aberrations. Comparison of 8,493 newborns (804,212 cells) from high-level natural radiation areas (dose rate >1.5 mGy/year) and 1,737 newborns (159,728 cells) from normal-level natural radiation areas (</=1.5 mGy/year) did not show any significant difference in the frequency of dicentrics, translocations, inversions or other types of aberrations known to be associated with radiation exposure. The cytogenetic studies were continued for constitutional anomalies using karyotype analysis, and scoring of 16,169 newborns has been completed. The overall frequency of constitutional anomalies was 4.95 +/- 0.55 per 1,000 newborns, which is comparable to the incidence reported in the literature. Within the limitations of sample size, the frequencies of total autosomal and sex aneuploids as well as structural anomalies were comparable between the high-level and normal-level natural radiation areas. A striking observation was the presence of rogue cells, the rarely occurring metaphases with a high level of chromosomal damage, which have not been reported previously among newborns.