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.

Immune status of people living in the Tande-Tande sub-village (Indonesia), an area with high indoor radon concentration.

On Earth, there are significant variations in terms of exposure to naturally occurring radiation among different areas. Radon, a naturally-occurring radioactive gas that is the primary cause of lung cancer in nonsmokers and the second most prevalent cause among smokers, poses a considerable risk. Indoor radon, in particular, constitutes the most substantial source of natural radiation to which individuals are exposed. This study assessed the immune status of a population chronically exposed to high indoor radon concentration in Indonesia. Fifty-seven subjects from the Tande-Tande sub-village (high indoor radon concentration area) were compared to fifty-three participants living in the Topoyo village (low concentration area). We contrasted the immunological conditions of these two populations by measuring levels of tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), interleukin-4 (IL-4), and IL-10 in serum. Moreover, we also measured levels of the nuclear factor kappa B (NF-κB), superoxide dismutase (SOD), glutathione peroxidase (GPX), and protein kinase B in its phosphorylated (pAkt) and non-phosphorylated form (Akt) in peripheral blood mononuclear cells (PBMCs) of a subset of participants (31 from each population). TNF-α, IFN-γ, and IL-4 levels in Tande-Tande sub-village inhabitants were significantly lower than those in the control group living in the Topoyo village (p = 0.001, p = 0.017, and p = 0.002). The concentration of IL-10 also tended to be lower in people living in the high indoor radon concentration area, but it did not differ significantly between Tande-Tande sub-village inhabitants and Topoyo inhabitants (p = 0.106). Protein levels of NF-κB, pAkt, and Akt in Tande-Tande sub-village inhabitants also did not differ significantly between Tande-Tande sub-village inhabitants and Topoyo inhabitants (p = 0.234, p = 0.210, and p = 0.657). Similarly, activities of SOD and GPX did not differ significantly between the two populations (p = 0.569 and p = 0.949). Overall, despite their chronic exposure to high indoor radon concentrations, our study revealed no increase in the levels of TNF-α, IFN-γ, IL-10, IL-4, SOD, and GPX in the inhabitants of Tande-Tande sub-village compared with people living in the Topoyo village. Furthermore, our study demonstrated no activation in the Akt pathway, as indicated by the pAkt/Akt ratio observed in PBMC lysates of individuals residing in the Tande-Tande sub-village.

γ-H2AX and phospho-ATM enzyme-linked immunosorbent assays as biodosimetry methods for radiation exposure assessment: a pilot study.

In the event of a radiological incident, a fast and accurate biological dosimetry (biodosimetry) method for evaluating people who have been potentially exposed to ionising radiation is crucial. Among the many biodosimetry methods available, the immunodetection of phosphorylated H2AX (γ-H2AX) stands as a promising method to be used in the triage of patients exposed to radiation. Currently, the most common way to measure γ-H2AX levels is through fluorescence microscopy. In this pilot study, we assessed the feasibility of using an enzyme-linked immunosorbent assay (ELISA) for quantifying γ-H2AX for biodosimetry purposes. Moreover, the usefulness of measuring phosphorylated ATM (pATM) levels through ELISA for biodosimetry was also evaluated. Blood samples were obtained from three male donors (38 y) and were irradiated with 60Co (0, 1, 2 and 6 Gy). Peripheral blood mononuclear cells (PBMCs) were isolated and lysed before measuring γ-H2AX, total H2AX protein and pATM using ELISA kits. The dicentric chromosome assay (DCA) using whole blood was also performed for comparison. Data from all donors at each dose were pooled before statistical analysis. The ratio of γ-H2AX/total H2AX and pATM levels increased in a radiation-dose-dependent manner. The average γ-H2AX/total H2AX ratios were 0.816 ± 0.219, 0.830 ± 0.685, 1.276 ± 1.151 and 1.606 ± 1.098, whereas the average levels of pATM were 59.359 ± 3.740, 63.366 ± 0.840, 66.273 ± 2.603 and 69.936 ± 4.439, in PBMCs exposed to 0, 1, 2 and 6 Gy, respectively. The linear-quadratic dose-response calibration curve for DCA was Y = 0.0017 (±0.0010) + 0.0251 (±0.0142) × D + 0.0342 (±0.0039) × D2  $\boldsymbol{Y}=\mathbf{0.0017}\left(\pm \mathbf{0.0010}\right)+\mathbf{0.0208}\left(\pm \mathbf{0.0218}\right)\times \boldsymbol{D}+\mathbf{0.0350}\left(\pm \mathbf{0.0050}\right)\times{\boldsymbol{D}}^{\mathbf{2}}$. Overall, despite a large variability in the ratio of γ-H2AX/total H2AX among donors, the present study revealed the suitability of using the ratio of γ-H2AX/total H2AX and pATM for biodosimetry. Still, more research with a larger group of subjects is necessary to construct a reliable calibration curve for the ratio of γ-H2AX/total H2AX and pATM levels for biodosimetry.

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.

Intestinal bacteria diversity of suckermouth catfish (Pterygoplichthys pardalis) in the Cd, Hg, and Pb contaminated Ciliwung River, Indonesia.

The contamination of aquatic environments with heavy metals poses a serious threat to fish, potentially leading to diseases or even death. Therefore, there is an urgent need for studies to investigate the adaptability of fish in heavy metal-contaminated environments. Several studies have explored the adaptability of suckermouth catfish (P. pardalis) to survive in the contaminated Ciliwung River. The findings obtained showed that the presence of intestinal bacteria helped these fish overcome the heavy metals in their intestines, thereby enabling the fish to survive. Analysis using the Next Generation Sequencing (NGS) technology has succeeded in identifying diversity of these bacteria in P. pardalis living in the Ciliwung River, which contaminated with Cd (0.3-1.6 ppm in the water & 0.9-1.6 ppm in the sediment), Hg (0.6-2 ppm in the water & 0.6-1.8 ppm in the sediment), and Pb (59.9-73.8 ppm in the water & 26.1-58.6 ppm in the sediment). Diversity index of intestinal bacteria in P. pardalis was relatively high, but it had a negative correlation with the presence of these contaminants. Actinobacteria, Firmicutes, and Proteobacteria were abundant in the intestines of P. pardalis from the upstream to downstream of the river, with an overall abundance range of 15-48%. Furthermore, Mycobacterium along with 6 other genera were identified as core intestinal bacteria. The presence of these bacterial communities in all the samples affected their survival in heavy metals-contaminated rivers. The fish's adaptability to live in this harsh environment indicated that it has the potential to be utilized as a bioremediator of heavy metals in river sediments.

Chromosome aberrations, micronucleus frequency, and catalase concentration in a population chronically exposed to high levels of radon.

PURPOSE: To deepen our knowledge on the effects of high levels of indoor radon exposure, we assessed the frequencies of unstable and stable chromosome aberrations and micronucleus (MN), as well as the concentration of an endogenous antioxidant (catalase, CAT), in blood samples of individuals chronically exposed to high indoor radon concentrations in Indonesia (Tande-Tande sub-village, Mamuju, West Sulawesi). Moreover, we also investigated the occurrence of a radio-adaptive response (RAR) in Tande-Tande sub-village inhabitants using the G2 MN assay. MATERIALS AND METHODS: The frequencies of dicentric (DC), acentric (AF), ring (R), and translocation (Tr) chromosomes in Tande-Tande inhabitants were compared to those in people living in a reference area with low levels of indoor radon levels (Topoyo village, Indonesia). The number of MN per 1000 binucleated cells (BNC) and CAT concentration per total protein was quantified and compared between groups. Lastly, we irradiated (2 Gy) phytohemagglutinin-stimulated samples in vitro and measured the frequency of MN to verify the occurrence of a RAR in Tande-Tande sub-village inhabitants. RESULTS AND CONCLUSION: The frequencies of DC, AF, and Tr did not differ between Tande-Tande inhabitants and control subjects (p = 0.350, 0.521, 0.597). The frequency of MN in Tande-Tande inhabitants was significantly lower than that in the control group (p = 0.006). Similarly, CAT concentration in Tande-Tande inhabitants was also significantly lower than that in the control population (p < 0.001). Significant negative correlations were identified for MN number and CAT concentration versus indoor radon concentration, annual effective dose, or cumulative dose both within groups and when all data were analyzed together. Our findings indicate that, despite the high indoor radon levels, Tande-Tande inhabitants are not under oxidative stress, since this group had lower CAT concentration and MN frequency than those in the control group. The negative correlation between MN frequency and indoor radon concentration, annual effective dose, and cumulative dose suggests the occurrence of an RAR phenomenon in Tande-Tande sub-village inhabitants. This interpretation is also supported by the results of the G2 MN assay, which revealed lower MN frequencies after in vitro irradiation of samples from Tande-Tande sub-village inhabitants than those in samples from the control group (p = 0.0069, for cumulative MN frequency; p = 0.0146, for radiation-induced MN only).

Microalgae Diversity in Interim Wet Storage of Spent Nuclear Fuel in Serpong, Indonesia.

The water quality in the interim wet storage of spent fuel (ISSF) needs to be monitored due to its function as a radiation shield. Water in ISSF pools must be free from microorganisms such as microalgae that live in a radioactive environment. Moreover, particular microalgae are capable of causing corrosion to stainless steel, which is a component of ISSF. Therefore, this study aims to determine the diversity of microalgae in the ISSF and those living in a radioactive environment, which cause corrosion. The microalgae were detected using the diversity and Palmer indices. The sampling of microalgae water was carried out by vertical filtration method at eight sites of ISSF. The results show that the diversity of microalgae (H') was low due to radiation exposure in pool water, hence, only specific species can survive. The evenness (J') of the microalgae was low, causing a high dominance index (C) value. Furthermore, the dominating species, namely, Chlorella sp. (Chlorophyceae), needs to be monitored because it has gamma radioresistance capabilities and can cause the corrosion of stainless steel.

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.