Quantification of histone H2AX phosphorylation in white blood cells induced by ex vivo gamma irradiation of whole blood by both flow cytometry and foci counting as a dose estimation in rapid triage.

A quick, reliable, and reproducible biological assay to distinguish individuals with possible life-threatening risk following radiological or nuclear incidents remains a quest in biodosimetry. In this paper, we examined the use of a γ-H2AX assay as an early dose estimation for rapid triage based on both flow cytometry and image analyses. In the experiment, whole blood from 11 donors was irradiated ex vivo inside a water phantom by gamma rays from Co-60 at 0.51 Gy/min. After the lysis of red blood cells, the white blood cells were collected for immunofluorescence labeling of γ-H2AX, CD45, and nuclear stained for signal collection and visualization. Analysis by flow cytometry showed that the relative γ-H2AX intensities of lymphocytes and granulocytes increased linearly with absorbed doses from 0 to 6 Gy with a large variation among individuals observed above 2 Gy. The relative γ-H2AX intensities of lymphocytes assessed by two different laboratories were highly correlated (ICC = 0.979). Using confocal microscopic images, γ-H2AX foci were observed to be discretely distributed inside the nuclei and to increase proportionally with doses from 0 to 2 Gy, whereas large plagues of merged foci appeared at 4 and 6 Gy, resulting in the saturation of foci counts above 4 Gy. The number of total foci per cell as well as the number of foci per plane were significantly different at 0 vs 1 and 2 vs 4 Gy doses (p < 0.01). Blind tests at 0.5 Gy and 1 Gy doses showed that dose estimation by flow cytometry had a mean absolute difference of less than 0.5 Gy from the actual value. In conclusion, while flow cytometry can provide a dose estimation with an uncertainty of 0.5 Gy at doses ≤ 1 Gy, foci counting can identify merged foci that are prominent at doses ≥ 4 Gy.

Occupational health risks among trichloroethylene-exposed workers in a clock manufacturing factory.

Trichloroethylene (TCE) is an important volatile organic compound once widely used in industry throughout the world. Occupational exposure to TCE can cause a number of health hazards such as allergic reactions and genetic damage. The purpose of this study was to evaluate occupational exposure to TCE, by analysis of the air in the breathing zone and of urine from workers employed in a clock manufacturing factory. A subjective symptom survey was conducted by using a self-administered questionnaire to evaluate the health hazards. Micronucleus (MN) frequency, based on the cytokinesis-block micronucleus assay (CBMN) in peripheral blood lymphocytes, (PBLs) was used as a biomarker for chromosome damage. A total of 244 participants, including 171 workers occupationally exposed to TCE and 73 non-exposed control employees, working mainly in office jobs in the same factory, were enrolled in this study. Analyses of airborne TCE concentrations in the workplace, and of urinary trichloroacetic acid (TCA) of the workers and controls, were performed by Gas Chromatography-Electron Capture Detector (GC-ECD) using the modified headspace technique. The average concentration of TCE in the workplace breathing zone was 27.83 ± 6.02 ppm. The average level of urinary TCA of the exposed workers and controls was 14.84 ± 1.62, 2.95 ± 0.28 mg/L. The frequency of MN/1000BN was 7.029 ± 0.39, significantly higher than for those in the control group (3.57 ± 0.31, p = 0.001). According to multiple linear regression analysis, the results indicated that urinary TCA levels correlated with the increased MN in exposed workers (r = 0.285, p < 0.001). The prevalence rate of subjective symptoms in the exposed group was 9.61-11.76 times higher than the rate of the non-exposed group (p < 0.001). It was found that skin (29.6%) and respiratory symptoms (21.1%) were the most frequent among the exposed workers. In conclusion, these results indicate that increased micronucleus frequency is associated with occupational trichloroethylene exposure. The use of TCE in the factory is threatening workers' health.