Evaluation of the action of LPSF/GQ-16 on cytokines and PPAR-γ gene expression after in vitro irradiation of peripheral blood mononuclear cells.

PURPOSE: Patients submitted to radiotherapy (RT) may present in their healthy tissues surrounding the treated tumor, some typical acute inflammatory reactions induced by ionizing radiation (IR). The manifestation of inflammatory processes is a result of exacerbation of the immune system, as a response to radiation exposure, and this can be a limiting factor for RT protocols. To counteract this, some thiazolidinediones, such as LPSF/GQ-16, may be useful for modulating the patient's radioinduced inflammatory response in normal tissues. In this context, the present work aims to evaluate the activity of LPSF/GQ-16 on the levels of cytokines and the expression of the gene PPARγ in mononuclear cells irradiated in vitro, to analyze the immunomodulatory activity of the molecule and its action on radiomitigation. MATERIALS AND METHODS: For this, blood samples from eight donors were collected and irradiated with 2 Gy, then the PBMC (peripheral blood mononuclear cells) were cultured and treated with LPSF/GQ-16. The levels of cytokines TNF-α, IFN-γ, IL-2 and IL-4 were quantified by CBA, while the genes of TNF-α, IFN-γ and PPARγ were analyzed by RT-PCR. RESULTS: LPSF/GQ-16 significantly reduced the expression of proinflammatory cytokines (IFN-γ and TNF-α) in irradiated and nonirradiated groups. There was no significant reduction of anti-inflammatory cytokines (IL-2 and IL-4) by LPSF/GQ-16. The mRNA expression of PPAR-γ, IFN-γ and TNF-α in the presence of LPSF/GQ-16 was higher in the nonirradiated sample. CONCLUSION: LPSF/GQ-16 showed effective activity after irradiation, with an important immunomodulatory activity in irradiated PBMCs.

α Espec-222 as a variant to the radon monitoring method using a solid state nuclear track.

Radon is important in indoor radiometry, where radiological impact is relevant. The improvement of low-cost methods makes it possible to maximize the monitored areas. Thus, a simple and robust monitoring device based on SSNTD CR-39 was designed. Detector conditions were standardized to reduce operational errors and increase productivity. The calibration factor by the slope method allowed efficiency greater than 93%. The monitoring system is satisfactory in terms of operation and performance, suitable for a wide range of radon monitoring situations.

Micronucleus assay for predicting side effects of radiotherapy for cervical cancer.

Radiotherapy (RT) is an important treatment for cervical cancer. The quality of life of patients undergoing RT may be compromised during and following treatment by nausea, diarrhea, vomiting, burns, erythema and fistula. Cytokinesis-block micronucleus (CBMN) assays may be useful for predicting adverse effects of RT for cancer. The CBMN test is easy to perform and is reproducible for screening subjects exposed to ionizing radiation. We investigated the use of the frequency of micronuclei (MN) from peripheral blood samples, irradiated in vitro, as a possible biomarker to predict the side effects of RT in patients with cervical cancer. We used 10 patients with cervical cancer receiving RT and chemotherapy. We found a strong relation between the frequency of MN and the appearance of acute side effects of RT for cervical cancer. We suggest that the methodology presented here may be useful for predicting side effects of RT for patients affected by cervical cancer and who have undergone chemotherapy.

Study of the Potential Radiomitigator Effect of Quercetin on Human Lymphocytes.

Several substances of synthetic and natural origin have been studied to determine their ability to protect the body from damage caused by ionizing radiation. Among these substances, quercetin has been shown to be a naturally occurring molecule with high radioprotective and radiomitigator potential due to its antioxidant properties. The objective of this work was to ascertain the potential radiomitigator effect of quercetin on chromosome aberration yield in lymphocytes of in vitro-irradiated human peripheral blood. At first, the DPPH (2,2-diphenyl-1-picryl-hydrazyl) radical capture test was performed to determine the antioxidant activity of quercetin and to select the concentrations to be tested. The blood was irradiated at doses of 2.5, 3.5, and 4.5 Gy and lymphocytes were cultured with quercetin at preselected concentrations of 37.5 and 75 µM. Then, the slides were prepared for scoring unstable chromosome aberrations (dicentrics, rings, and fragments). The results showed that the lymphocytes irradiated and later exposed to quercetin presented a lower frequency of chromosomal alterations compared to the control sample which was irradiated and not exposed to quercetin. The results suggest a potential radiomitigator effect of the flavonoid quercetin on human lymphocytes exposed, in vitro, to ionizing radiation. This effect may be related to decrease in the release of cytokines (INF-γ, PGE2, IL-1ß, IL6, IL-8) involved in the proinflammatory processes as well as downregulation of NF-kB and reduction of expression TGF-ß.

Evaluation of Pb-210 in urine and frequency of micronuclei in exfoliated cells as indicators of exposure to cigarettes.

This study aimed at analyzing the frequency of micronuclei (MN) in exfoliated cells as well as the levels of Pb-210 in urine samples to evaluate the association between the smoking habit and toxic stress of transitional epithelial cells. The frequency of MN was scored from Giemsa-stained slides while exchange resin and beta counting techniques were employed to measure the concentrations of this radioisotope. Urine samples of smokers had levels of Pb-210 up to 158.65 mBq L-1. For nonsmokers, the median was below the detection limit (45 mBq L-1). The analyses of mononucleated cells showed a significant increase of the frequency of MN in smokers when compared to nonsmokers. Statistical tests showed a tight relation between the cigarette consumption and the increase of the frequency of MN, rather than with the levels of Pb-210 present in smoke particles. The results indicate the usefulness of the methodology for the evaluation of human health risks related to chronic contamination with Pb-210.

Image processing in biodosimetry: A proposal of a generic free software platform.

The scoring of chromosome aberrations is the most reliable biological method for evaluating individual exposure to ionizing radiation. However, microscopic analyses of chromosome human metaphases, generally employed to identify aberrations mainly dicentrics (chromosome with two centromeres), is a laborious task. This method is time consuming and its application in biological dosimetry would be almost impossible in case of a large scale radiation incidents. In this project, a generic software was enhanced for automatic chromosome image processing from a framework originally developed for the Framework V project Simbio, of the European Union for applications in the area of source localization from electroencephalographic signals. The platforms capability is demonstrated by a study comparing automatic segmentation strategies of chromosomes from microscopic images.

p53 protein expression levels as bioindicator of individual exposure to ionizing radiation by flow cytometry.

Ionizing radiation (IR) can cause various lesions in DNA, which induce the increase of p53 expression levels in order to repair radiation induced damage. Thus, the correlation between the increase of p53 expression and an irradiation may constitute a fast and powerful method of individual monitoring in cases of accidental or suspected exposures to IR. In this context, the aim of this research was to evaluate changes in lymphocyte p53 expression levels, based on flow cytometry, after in vitro irradiation of peripheral blood samples. For the measurement of such expression levels of p53 protein, an investigation was carried out in order to establish a methodology of analysis based on flow cytometry. Hence, relationships among levels of expression of p53 protein with the absorbed dose have been verified. The results presented in this report emphasized flow cytometry as an important tool for the fast evaluation of p53 protein expression levels as bioindicator of individual exposure to acute ionizing radiation.