Integrative proteomic and microRNA analysis of primary human coronary artery endothelial cells exposed to low-dose gamma radiation.

High doses of ionising radiation significantly increase the risk of cardiovascular disease (CVD), the vascular endothelium representing one of the main targets. Whether radiation doses lower than 500 mGy induce cardiovascular damage is controversial. The aim of this study was to investigate radiation-induced expression changes on protein and microRNA (miRNA) level in primary human coronary artery endothelial cells after a single 200 mGy radiation dose (Co-60). Using a multiplex gel-based proteomics technology (2D-DIGE), we identified 28 deregulated proteins showing more than ±1.5-fold expression change in comparison with non-exposed cells. A great majority of the proteins showed up-regulation. Bioinformatics analysis indicated "cellular assembly and organisation, cellular function and maintenance and molecular transport" as the most significant radiation-responsive network. Caspase-3, a central regulator of this network, was confirmed to be up-regulated using immunoblotting. We also analysed radiation-induced alterations in the level of six miRNAs known to play a role either in CVD or in radiation response. The expression of miR-21 and miR-146b showed significant radiation-induced deregulation. Using miRNA target prediction, three proteins found differentially expressed in this study were identified as putative candidates for miR-21 regulation. A negative correlation was observed between miR-21 levels and the predicted target proteins, desmoglein 1, phosphoglucomutase and target of Myb protein. This study shows for the first time that a low-dose exposure has a significant impact on miRNA expression that is directly related to protein expression alterations. The data presented here may facilitate the discovery of low-dose biomarkers of radiation-induced cardiovascular damage.

Simulated and measured dose response characteristics of detectors used for CT dosimetry.

A CT-SD16 semiconductor detector was calibrated in terms of the computed tomography (CT) air kerma index for the integration length L = 100 mm, C(K,PMMA,100), in the cylindrical CT head and body dosimetry phantoms using a DCT10 pencil ionization chamber as a reference instrument. Using IEC RQT 9 120-kV x-ray radiation quality and 25-62.5 mm nominal beam widths free in air, a C(K,PMMA,100)(DCT10)/C(K,PMMA,100)(CT-SD16) ratio of 0.97 was observed, while in the centre of 300 mm long CT head and body dosimetry phantoms, C(K,PMMA,100)(DCT10)/C(K,PMMA,100)(CT-SD16) ratios ranged from 1.02 to 1.09. Using IEC RQT 8-10 radiation qualities free in air, Monte Carlo simulated dose response characteristics of CT-SD16 and DCT10 were comparable with those obtained from the measurements. Simulations were also used to determine C(K,PMMA,100)(DCT10) in the centre of the CT head and body phantoms. At IEC RQT 9 and 25-62.5 mm nominal beam widths, the relative values of the simulated dose agreed with the measured values within 2-10% for the head and body phantoms, respectively. A k(q) correction factor between dose measurements in the phantom and free in air was determined for the model of DCT10 and for the real detectors using measured C(K,PMMA,100) data. Simulations were performed using the EGSnrc CAVRZ code.

Low-dose irradiation causes rapid alterations to the proteome of the human endothelial cell line EA.hy926.

High doses of ionising radiation damage the heart by an as yet unknown mechanism. A concern for radiological protection is the recent epidemiological data indicating that doses as low as 100-500 mGy may induce cardiac damage. The aim of this study was to identify potential molecular targets and/or mechanisms involved in the pathogenesis of low-dose radiation-induced cardiovascular disease. The vascular endothelium plays a pivotal role in the regulation of cardiac function and is therefore a potential target tissue. We report here that low-dose radiation induced rapid and time-dependent changes in the cytoplasmic proteome of the human endothelial cell line EA.hy926. The proteomes were investigated at 4 and 24 h after irradiation at two different dose rates (Co-60 gamma ray total dose 200 mGy; 20 mGy/min and 190 mGy/min) using 2D-DIGE technology. Differentially expressed proteins were identified, after in-gel trypsin digestion, by MALDI-TOF/TOF tandem mass spectrometry, and peptide mass fingerprint analyses. We identified 15 significantly differentially expressed proteins, of which 10 were up-regulated and 5 down-regulated, with more than ±1.5-fold difference compared with unexposed cells. Pathways influenced by the low-dose exposures included the Ran and RhoA pathways, fatty acid metabolism and stress response.

Simulated and measured spectral characteristics of 137Cs reference radiation beams.

137Cs sources are widely used in calibrations of radiation protection dosimeters and doserate meters. In these calibrations, conversion coefficients from the air kerma to various dose equivalent quantities are needed. These conversion quantities are functions of photon energy. In this work, Monte Carlo simulations and measurements are used for quantifying the energy distributions of the photon fluence from two 137Cs sources used for calibration purposes. The results indicate that the standard set of conversion coefficients, obtained from a monoenergetic spectrum, do not sufficiently take into account the scattered radiation (most of which is scattered by the source itself): For the large (14x15.6 mm) and small (3.5x3 mm) 137Cs sources, the simulated values for the conversion coefficients H'(0.07)/Ka and H*(10)/Ka (which were nearly equal for the both quantities) showed around 1.0 and 1.5% deviations from the ISO standard value 1.20 (for the large and small sources, respectively). Similarly, the conversion coefficient Hp(10)/Ka showed around 1.5 and 2.5% deviations from the ISO standard value 1.21 for the large and small 137Cs sources, respectively, whereas Hp(0.07)/Ka had the values 1.22 and 1.23. The amplitude of the variation of the conversion coefficients due to varying 137Cs source size did not exceed 1%. Thus, the overall uncertainty (with coverage factor k=2) due to varying source size for the average values of the conversion coefficients can be estimated as low as 0.6%.

Portable TL dosemeter--ESD phantom combination for chest and lumbar spine radiography.

A thermoluminescence dosemeter (TLD)-entrance surface dose (ESD) phantom combination was calibrated in terms of air-kerma in IEC RQR X-ray radiation qualities between 50 and 150 kV. The ESD phantom was designed and constructed as a part of the work. With the combination, air-kermas were measured for four radiological examinations (two chest and two lumbar spine examinations in two hospitals), and converted to ESDa using Monte-Carlo calculated BSF data tabulated for different tube voltages, filtrations and beam diameters at 1 m distance. The results agreed with the ionisation-chamber measurements within the reported overall uncertainty of the TLD method. In the calibration, the ESD phantom can be replaced by the ISO water slab phantom unaffecting the reported overall uncertainty. Backscatter-related parameters for the ISO water slab phantom and the newly designed ESD phantom were determined for the IEC RQR qualities used in the secondary standard dosimetry laboratory of STUK at 1 m distance, including an approximate Hp(10)/Hp(0.07) ratio.

Determination of conversion factors from air kerma to operational dose equivalent quantities for low-energy X-ray spectra.

The conversion coefficients from air kerma to ICRU operational dose equivalent quantities for STUK's realisation of the X-radiation qualities N-15 to N-60 of the ISO narrow (N) spectrum series were determined by utilising X-ray spectrum measurements. The pulse-height spectra were measured using a planar high-purity germanium spectrometer and unfolded to fluence spectra using Monte Carlo generated data of the spectrometer response. To verify the measuring and unfolding method, the first and second half-value layers and the air kerma rate were calculated from the fluence spectra and compared with the values measured using an ionisation chamber. For each radiation quality, the spectrum was characterised by the parameters given in ISO 4037-1. The conversion coefficients from the air kerma to the ICRU operational quantities Hp(10), Hp(0.07), H'(0.07) and H*(10) were calculated using monoenergetic conversion coefficients at zero angle of incidence. The results are discussed with respect to ISO 4037-4, and compared with published results for low-energy X-ray spectra.

Design, spectrum measurements and simulations for a 238Pu alpha-particle irradiator for bystander effect and genomic instability experiments.

Design, spectrum measurements and simulations for an alpha-particle irradiator for bystander effect and genomic instability experiments are presented. Measured alpha-particle energy spectra were used to confirm the characteristics of the source of the irradiator specified by the manufacturer of the source. The spectra were measured in vacuum with a high-resolution spectrometer and simulated with an AASI Monte Carlo code. As a next step, we simulated alpha-particle energy spectra at the target plane of the irradiator for three different source-to-target distances. In these simulations, helium was used as the medium between the source and the exit window of the irradiator; its pressure and temperature corresponded to those of the ambient air. Mean energies and full-widths at half-maximum (FWHM) were calculated for the three different helium gas tracks.