Sornjarod Oonsiri

LINE-1 methylation status of endogenous DNA double-strand breaks

DNA methylation and the repair of DNA double-strand breaks (DSBs) are important processes for maintaining genomic integrity. Although DSBs can be produced by numerous agents, they also occur spontaneously as endogenous DSBs (EDSBs). In this study, we evaluated the methylation status of EDSBs to determine if there is a connection between DNA methylation and EDSBs. We utilized interspersed repetitive sequence polymerase chain reaction (PCR), ligation-mediated PCR and combined bisulfite restriction analysis to examine the extent of EDSBs and methylation at long interspersed nuclear element-1 (LINE-1) sequences nearby EDSBs. We tested normal white blood cells and several cell lines derived from epithelial cancers and leukemias. Significant levels of EDSBs were detectable in all cell types. EDSBs were also found in both replicating and non-replicating cells. We found that EDSBs contain higher levels of methylation than the cellular genome. This hypermethylation is replication independent and the methylation was present in the genome at the location prior to the DNA DSB. The differences in methylation levels between EDSBs and the rest of the genome suggests that EDSBs are differentially processed, by production, end-modification, or repair, depending on the DNA methylation status.

Optimal sensitometric curves of Kodak EDR2 film for dynamic intensity modulated radiation therapy verification.

Purpose: To investigate the optimal sensitometric curves of extended dose range (EDR2) radiographic film in terms of depth, field size, dose range and processing conditions for dynamic intensity modulated radiation therapy (IMRT) dosimetry verification with 6 MV X-ray beams. Materials and methods: A Varian Clinac 23 EX linear accelerator with 6 MV X-ray beam was used to study the response of Kodak EDR2 film. Measurements were performed at depths of 5, 10 and 15 cm in MedTec virtual water phantom and with field sizes of 2x2, 3x3, 10x10 and 15x15 cm2. Doses ranging from 20 to 450 cGy were used. The film was developed with the Kodak RP X-OMAT Model M6B automatic film processor. Film response was measured with the Vidar model VXR-16 scanner. Sensitometric curves were applied to the dose profiles measured with film at 5 cm in the virtual water phantom with field sizes of 2x2 and 10x10 cm2 and compared with ion chamber data. Scanditronix/Wellhofer OmniProTM IMRT software was used for the evaluation of the IMRT plan calculated by Eclipse treatment planning. Results: Investigation of the reproducibility and accuracy of the film responses, which depend mainly on the film processor, was carried out by irradiating one film nine times with doses of 20 to 450 cGy. A maximum standard deviation of 4.9% was found which decreased to 1.9% for doses between 20 and 200 cGy. The sensitometric curves for various field sizes at fixed depth showed a maximum difference of 4.2% between 2x2 and 15x15 cm2 at 5 cm depth with a dose of 450 cGy. The shallow depth tended to show a greater effect of field size responses than the deeper depths. The sensitometric curves for various depths at fixed field size showed slightly different film responses; the difference due to depth was within 1.8% for all field sizes studied. Both field size and depth effect were reduced when the doses were lower than 450 cGy. The difference was within 2.5% in the dose range from 20 to 300 cGy for all field sizes and depths studied. Dose profiles measured with EDR2 film were consistent with those measured with an ion chamber. The optimal sensitometric curve was acquired by irradiating film at a depth of 5 cm with doses ranging from 20 to 450 cGy with a 3×3 cm2 multileaf collimator. The optimal sensitometric curve allowed accurate determination of the absolute dose distribution. In almost 200 cases of dynamic IMRT plan verification with EDR2 film, the difference between measured and calculated dose was generally less than 3% and with 3 mm distance to agreement when using gamma value verification. Conclusion: EDR2 film can be used for accurate verification of composite isodose distributions of dynamic IMRT when the optimal sensitometric curve has been established.

Measurements of patient's setup variation in intensity-modulated radiation therapy of head and neck cancer using electronic portal imaging device.

Purpose: To measure the interfraction setup variation of patient undergoing intensity-modulated radiation therapy (IMRT) of head and neck cancer. The data was used to define adequate treatment CTV-to-PTV margin. Materials and methods: During March to September 2006, data was collected from 9 head and neck cancer patients treated with dynamic IMRT using 6 MV X-ray beam from Varian Clinac 23EX. Weekly portal images of setup fields which were anterior-posterior and lateral portal images were acquired for each patient with an amorphous silicon EPID, Varian aS500. These images were matched with the reference image from Varian Acuity simulator using the Varis vision software (Version 7.3.10). Six anatomical landmarks were selected for comparison. The displacement of portal image from the reference image was recorded in X (Left-Right, L-R), Y (Superior-Inferior, S-I) direction for anterior field and Z (Anterior-Posterior, A-P), Y (S-I) direction for lateral field. The systematic and random error for individual and population were calculated. Then the population-based margins were obtained. Results: A total of 135 images (27 simulation images and 108 portal images) and 405 match points was evaluated. The systematic error ranged from 0 to 7.5 mm and the random error ranged from 0.3 to 4.8 mm for all directions. The population-based margin ranged from 2.3 to 4.5 mm (L-R), 3.5 to 4.9 mm (S-I) for anterior field and 3.4 to 4.7 mm (A-P), 2.6 to 3.7 mm (S-I) for the lateral field. These margins were comparable to the margin that was prescribed at the King Chulalongkorn Memorial Hospital (5-10 mm) for head and neck cancer. Conclusion: The population-based margin is less than 5 mm, thus the margin provides sufficient coverage for all of the patients.