Exposure to ionizing radiation and hyperthermia results in important biological consequences, e.g. cell death, chromosomal aberrations, mutations, and DNA strand breaks. There is good evidence that the nucleus, specifically cellular DNA, is the principal target for radiation-induced cell lethality. DNA double-strand breaks (DSBs) are considered to be the most serious type of DNA damage induced by ionizing radiation. On the other hand, verifiable mechanisms which can lead to heat-induced cell death are damage to the plasma membrane and/or inactivation of heat-labile proteins caused by protein denaturation and subsequent aggregation. Recently, several reports have suggested that DSBs can be induced after hyperthermia because heat-induced phosphorylated histone H2AX (γ-H2AX) foci formation can be observed in several mammalian cell lines. In mammalian cells, DSBs are repaired primarily through two distinct and complementary mechanisms: non-homologous end joining (NHEJ), and homologous recombination (HR) or homology-directed repair (HDR). DNA-dependent protein kinase (DNA-PK) and ataxia-telangiectasia mutated (ATM) are key players in the initiation of DSB repair and phosphorylate and/or activate many substrates, including themselves. These phosphorylated substrates have important roles in the functioning of cell cycle checkpoints and in cell death, as well as in DSB repair. Apoptotic cell death is a crucial cell suicide mechanism during development and in the defense of homeostasis. If DSBs are unrepaired or misrepaired, apoptosis is a very important system which can protect an organism against carcinogenesis. This paper reviews recently obtained results and current topics concerning the role of DNA-PK and ATM in heat- or radiation-induced apoptotic cell death.

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Artemis is a structure-specific endonuclease when associated with and phosphorylated by DNA-dependent protein kinase catalytic subunit. This structure-specific endonuclease is responsible for the resolution of hairpin coding ends in V(D)J recombination. In DNA double-strand break repair, Artemis is implicated in the end-processing step of the non-homologous end-joining (NHEJ) pathway. Recently, we have demonstrated that the involvement of Artemis in NHEJ depends on the type of DNA damage. Interestingly, recent evidence suggests that the end-processing activity is not the only function of Artemis. Indeed, Artemis is rapidly phosphorylated by ataxia telangiectasia mutated in response to DNA damage, and such phosphorylation of Artemis appears to be involved in the regulation of cell cycle checkpoints. These findings suggest that Artemis is a multifunctional protein participating in the maintenance of genome integrity at two distinct levels; one at the end processing step of NHEJ, and the other at the signaling pathway of cell cycle regulation. Therefore, understanding Artemis function may give us profound insights into the DNA repair network. In this review, we summarize the functions and regulation of Artemis.

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The effects of topical application of recombinant human epidermal growth factor (rhEGF) on wound healing and the recurrence of radiodermatitis were assessed in the irradiated skin of BALB/c Nu/Nu mice. Mice irradiated with 45 Gy of radiation were divided into 5 groups and treated with 10, 50, and 100 μg/g rhEGF ointment, vehicle alone, or no treatment (control) for 6 months. Wounds were observed initially in all groups and complete healing time (HT₁₀₀) for initial wound repair did not differ significantly among groups. However, the rate of recurrence over 6 months was significantly lower in the EGF-treated groups than in the control group (p < 0.05). Histological examination showed that treatment with the optimum dose of EGF (50 μg/g) accelerated normal wound healing when compared with the higher dose of EGF (100 μg/g), vehicle alone, or no treatment, with the latter group showing irregular epidermal thickness, poor definition of epidermis and dermis, and unstable dermal structure. Collagen distribution was also significantly increased in mice treated with 50 μg/g rhEGF (p < 0.05) compared with the control or vehicle-treated group. Taken together, these results indicate that treatment with exogenous EGF (50 μg/g dose) can enhance radiation-induced wound repair while preserving structural tissue stability and preventing the recurrence of radiodermatitis.

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Purpose: To elucidate the mechanism underlying the in vivo radioprotection activity by Zn-containing, heat-treated Saccharomyces cerevisiae yeast (Zn-yeast). Materials and Methods: Zn-yeast suspension was administered into C3H/He mice immediately after whole body irradiation (WBI) at 7.5 Gy. Bone marrow was extracted from the mice 6 hours after irradiation and analyzed on a microarray. Expression changes in the candidate responsive genes differentially expressed in treated mice were re-examined by qRT-PCR. The bone marrow was also examined pathologically at 6 h, 3, 7, and 14 days postirradiation. Results: Thirty-six genes, including Edn1 and Agpt2, were identified as candidate responsive genes in irradiated mouse bone marrow treated with Zn-yeast by showing a greater than three-fold change compared with control (no irradiation and no Zn-yeast) mice. The expressions of Cdkn1a, Bax, and Ccng, which are well known as radioresponsive genes, were upregulated in WBI mice and Zn-yeast treated WBI mice. Pathological examination showed the newly formed microvessels lined with endothelial cells, and small round hematopoietic cells around vessels in bone marrow matrix of mice administered with Zn-yeast after WBI, while whole-body irradiated mice developed fatty bone marrow within 2 weeks after irradiation. Conclusion: This study identified a possible mechanism for the postirradiation protection conferred by Zn-yeast. The protective effect of Zn-yeast against WBI is related to maintaining the bone marrow microenvironment, including targeting endothelial cells and cytokine release.

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Vascular endothelial cells are very sensitive to ionizing radiation, and it is important to develop effective prevent agents and measures in radiation exposure protection. In the present study, the protective effects of atorvastatin on irradiated human umbilical vein endothelial cells (HUVEC) and the possible mechanisms were explored. Cultured HUVEC were treated by atorvastatin at a final concentration of 10 μ mol/ml for 10 minutes, and then irradiated at a dose of 2 Gy or 25 Gy. Twenty-four hours after irradiation, apoptosis of HUVEC was monitored by flow cytometry, and the expression of thrombomodulin (TM) and protein C activation in HUVEC was respectively assessed by flow cytometry and spectrophotometry. After treatment with atorvastatin for 24 h, the rate of cell apoptosis decreased by 6% and 16% in cells irradiated with 2 Gy and 25 Gy, respectively. TM expression increased by 77%, 59%, and 61% in untreated cells, 2 Gy irradiation-treated cells, and 25 Gy irradiation-treated cells, respectively. The protein C levels in 2 Gy and 25 Gy irradiation-treated cells were reduced by 23% and 34% when compared with untreated cells, but up-regulated by 79% and 76% when compared with cells which were irradiated and treated with atorvastatin. In conclusion, these data indicate that atorvastatin exerts protective effects on irradiated HUVEC by reducing apoptosis by up-regulating TM expression and enhancing protein C activation in irradiated HUVEC.

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This study was to investigate whether rhEGF protects radiation induced intestine injury without compromising antitumor effect of radiation in murine system. A radiation induced intestinal injury model was established in mice by whole body irradiation. Using this model, 4 groups were set; control, rhEGF (100 μg/kg intraperitoneally), radiation (10 Gy), and a combination (rhEGF and radiation). The level of apoptosis and proliferation were analyzed by TUNEL assay and proliferation cell nuclear antigen (PCNA) immunohistochemical staining, respectively, as well as observation of survival and body weight change. A tumor growth delay assay was performed using murine syngeneic tumors; one radioresistant tumor, HCa-I and one radiosensitive tumor, MCa-K. In the radiation induced intestinal injury model, the 10 Gy group had significantly more weight loss with less number of crypt cells and higher apoptosis than the 8 Gy group. Using 10 Gy model, radioprotective effect of rhEGF was tested. Addition of rhEGF improved not only the body weight loss but also survival following radiation. It also induced suppression of apoptosis as well as increase of PCNA expression and recovery of villi. rhEGF did not enhance the tumor growth after radiation exposure in the tested tumors. These findings suggest that combination of exogenous rhEGF and radiation can be a new anticancer strategy by protecting radiation-induced intestinal injury without alleviating antitumor effect of radiation.

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Although the use of small segments in step and shoot IMRT provides better dose distribution, extremely small segments decrease treatment accuracy. The purpose of this study was to determine the optimum minimum segment size (MSS) in two-step optimization in prostate step and shoot IMRT with regard to both planning quality and dosimetric accuracy. The XiO treatment planning system and Oncor Impression Plus were used. Results showed that the difference in homogeneity index (HI), defined as the ratio of maximum to minimum doses for planning target volume, between the MSS 1.0 cm and 1.5 cm plans, and 2.0 cm plans, was 0.1%, and 9.6%, respectively. With regard to V107 of PTV, the volume receiving 107% of the prescribed dose of the PTV, the difference between MSS 1.0 cm and 1.5 cm was 2%. However, the value of the MSS 2.0 cm or greater plans was more than 2.5-fold that of the MSS 1.0 cm plan. With regard to maximum rectal dose, a significant difference was seen between the MSS 1.5 cm and 2.0 cm plans, whereas no significant difference was seen between the MSS 1.0 cm and 1.5 cm plans. Composite plan verification revealed a greater than 5% dose difference between planned and measured dose in many regions with the MSS 1.0 cm plan, but in only limited regions in the MSS 1.5 cm plan. Our data suggest that the MSS should be determined with regard to both planning quality and dosimetric accuracy.

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In the present study, the prostate medium determined from the CT images of 149 patients was developed. The dosimetric parameters such as Λ, g_L(r) and F(r, θ) used in TG-43U1-based calculation for an iodine-125 (¹²⁵I) brachytherapy-source were examined using Monte Carlo code Geant4. Clinical dosimetry parameters such as the D₉₀ were evaluated among a subgroup of 50 randomly selected patients who had been treated with permanent brachytherapy between January 2008 and December 2008 at the Tokyo Medical Center. The results show a slight difference in the dose rate constant Λ (within 1.0%). The radial dose function g_L(r) exhibits a prominent difference in the region over 3 cm, and this difference is maintained within 2.9% in the region close to the source. The calculated values of F(r, θ) for the prostate medium were similar to values for water (within 1%), except in the longitudinal axis. A comparison of D₉₀ values shows a systematic dose overestimation of 2.8 ± 0.7 Gy in water, where the distribution of the differences can be seen with a spread of 1.8 ± 0.3% compared to that in prostate medium. It was concluded that the introduction of any kind of tissue correction for the TG-43U1-based calculation was not necessary to allow for the differences in elemental compositions and densities between water and prostate medium. PACS number: 87.00.00; 87.55.dk; 87.55.K-; 87.56.B-

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This paper provides quantitative structure activity relationship (QSAR) models for predicting the radiosensitization effectiveness of nitroimidazole compounds. A new method, combining a heuristic method and projection pursuit regression, was used to build an advanced QSAR model. Compared to the conventional multi-linear regression model, this model showed better predictive ability and reliability, with the values of regression coefficient (R²) and root mean square error (RMSE) 0.92 and 0.18 for the training set and 0.90 and 0.17 for the test set, respectively. The provided models were useful tools to predict the radiosensitization effectiveness of nitroimidazole compounds. Also, the new finding descriptors derived from this study will help us to facilitate the design of new radiation sensitizers with better activities.

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Deficiencies in repair cells and infection are two of the main factors that can hinder the process of wound healing. In the present study, we investigated the ability of human beta-defensin-2 (hBD2) genetically modified dermal multipotent stem cells (dMSCs) to accelerate the healing irradiated wounds complicated by infections. An hBD2 adenovirus expression vector (Adv-hBD2) was firstly constructed and used to infect dMSCs. The antibacterial activity of the supernatant was determined by Kirby-Bauer method and macrodilution broth assay. Time to complete wound healing, residual percentage of wound area, and the number of bacteria under the scar were measured to assess the effects of Adv-hBD2-infected dMSC transplantation on the healing of irradiated wounds complicated by Pseudomonas aeruginosa infection. Results showed that the supernatant from Adv-hBD2-infected dMSCs had obvious antibacterial effects. Transplantation of Adv-hBD2-infected dMSCs killed bacteria in the wound. The complete wound healing time was 19.8 ± 0.45 days, which was significantly shorter than in the control groups (P < 0.05). From 14 days after transplantation, the residual wound area was smaller in the experimental group than in the control groups (P < 0.05). In conculsion, we found that transplantation of hBD2 genetically modified dMSCs accelerated the healing of wounds complicated by P. aeruginosa infection in whole body irradiated rats.

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The purpose of this study is to compare three-dimensional conformal radiotherapy (3D-CRT) plans in a setting of salvage radiotherapy after radical prostatectomy (RP) and to simulate whether dose escalation is possible with the most adequate 3D-CRT technique. This study included consecutive 10 patients underwent salvage radiotherapy (RT) for biochemical relapse of prostate cancer after RP. Normal structures included the rectum, bladder, and femoral head. For each patient, four different treatment plans including four fields RT (4F-RT), dynamic conformal arc radiotherapy (DCAT), six fields RT (6F-RT), and DCAT with rectum hollow-out technique (DCAT-HO), were created to entire the prostate bed. The parameters of the maximum and mean doses received by organs at risk (OAR), target coverage, dose homogeneity for the planning target volume (PTV) were compared. All plans were considered to be clinically tolerable for PTV coverage and dose homogeneity. The rectum sparing at the high dose area for DCAT-HO was considered to be the most superior to those for other three techniques by comparison of the dose delivered to a 1%, 5%, and 10% volume of the rectum. In the simulation of dose escalation to 70 Gy with DCAT-HO, OAR met a requirement of the dose-volume constraints. However, in the simulation of dose escalation to 72 Gy, the rectum that receives 60 to 65 Gy and bladder that receives 65 Gy exceeded the optimal dose-volume constraints. DCAT-HO was considered to be one of the most appropriate techniques in 3D-CRT if dose escalation to 70 Gy might be needed in a setting of salvage RT after RP in the future.

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World Health Organization (WHO) grade 3 glioma is one of the common brain tumors and has three main histological subtypes, including anaplastic astrocytoma (AA), anaplastic oligoastrocytoma (AOA) and anaplastic oligodendroglioma (AO). However, most previous studies have considered AOA and AO as one group because of the difficult differential diagnosis between AOA and AO. Therefore the prognostic difference among patients with these histological subtypes has been unclear. In this study, 68 patients with histologically proven WHO grade 3 glioma, consecutively received postoperative radiotherapy at the Gunma University Hospital, Japan, between 1983 and 2005, were investigated to assess the impact of histological subtype on the survival. The number of AA, AOA and AO patients was 41, 16 and 11, respectively. The mean and median follow-up periods were 72 and 48 months, respectively. The number of patients treated with gross total resection, partial resection and biopsy was 14, 38 and 16, respectively. The mean and median radiation doses were 58 ± 5 Gy and 60 Gy, respectively. The 5-year overall survival rates of AA, AOA and AO were 21%, 38% and 80%, and median survival period were 16 months, 58 months and not reached, respectively. Univariate analysis showed that the histological subtype (P < 0.01) and extent of surgery (P < 0.01) were significant prognostic factors for survival. Selective comparison showed that overall survival of patients with AA was significantly worse than for those with AOA (P = 0.01) and AO (P < 0.01). The overall survival of patients with AO was better than for those with AOA; however, the difference was not statistically significant (P = 0.14). Multivariate analysis demonstrated that histological subtype, age and extent of surgery were the significant independent variable for survival (P < 0.01, P < 0.01 and P = 0.04). In our study, histological subtype was one of the most important prognostic factors of WHO grade 3 glioma.

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Concurrent chemotherapy with radiotherapy (CCRT) has been applied for the treatment of advanced stage of head and neck cancer patients. However CCRT is associated with several complications including mucositis, dermatitis, stomatitis, etc. This study was conducted to evaluate the therapeutic effect of systemically administrated recombinant human epidermal growth factor (rhEGF) in CCRT-induced oral mucositis in a mouse model. Oral mucositis was induced in male BALB/c mice through combination treatment with cisplatin (11 mg/kg, i.p.) and irradiation (17 Gy) of the head and neck area. rhEGF (1.0 mg/kg/day for consecutive 3 days) was administered systemically, and the therapeutic effect was determined by histological evaluation of the oral mucosa. To elucidate optimal dose of rhEGF on CCRT-induced mucositis, various concentrations (0.04–3 mg/kg) of rhEGF were injected for 3 days. Systemic rhEGF administration accelerated the recovery of body weight. Histologically, rhEGF-treated mice showed significantly increased epithelial cell layer thickness, basal cell number, and expression of Ki-67 compared to control mice. Most effective dose was 1 mg/kg among other doses tested. Systemic administration of 1 mg/kg of rhEGF reduces the severity of oral mucositis induced by CCRT in a mouse model, suggesting that rhEGF can be used for treating CCRT-induced mucositis during the cancer treatment.

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Head and neck cancer is treated mainly with surgery and radiotherapy. Xerostomia and mucositis are common adverse effects of radiation therapy. One of the strategies aimed at decreasing radiation toxicity is the use of radioprotective agents, such as amifostine. We previously reported that radio induced apoptosis of peripheral blood lymphocytes was statistically associated with normal tissue toxicity in the form of severe xerostomia. The aim of the present study was to explore the effects of amifostine on the radiation-induced apoptosis of peripheral blood lymphocytes from patients suffering head and neck cancer. Eighteen consecutive patients with squamous cell carcinoma of the head and neck were included in the study. Peripheral blood lymphocytes were isolated before and after the treatment with amifostine. Then, cells were irradiated at 0, 1, 2 and 8 Gy during 24 hours. Apoptosis was measured by flow cytometry using annexin V/propidium iodide. As expected, radio-induced apoptosis values fitted to a semi logarithmic equation as follows: RIA = β ln(Gy) + α. The administration of amifostine prior to radiation therapy modulates radio-induced apoptosis of peripheral blood lymphocytes: 13.68 vs. 13.37 (P = 0.027), 19.11 vs. 17.64 (P = 0.001) and 30.70 vs. 28.84 (P = 0.001), before and after the administration of the drug for 1, 2 and 8 Gy respectively. α and β decreased significantly after the administration of the drug: 13.58 vs. 12.99 (P = 0.009) and 8.21 vs. 7.53 (P = 0.017), respectively. Our results provide new information about the biological actions of amifostine in vivo.

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The aim of this study was to test the hypothesis that the "radical pair mechanism" (magnetic field effect on recombination rate of radical pairs) explains our previous findings indicating that 50 Hz magnetic fields (MF) of about 100 μT modify biological responses to ultraviolet (UV) radiation. In the present study, the effects of 50 Hz MF on cellular oxidative processes induced by UV radiation were investigated. Murine L929 fibroblast cells were exposed to 50 Hz MF of 100 or 300 μT during a 1-h UV exposure or for 24 h before it. The decay kinetics of oxidative reactions were analysed by measuring ultraweak chemiluminescence (photon emissions) of the exposed cells by scintillation counter in the out-of-coincidence mode. No significant MF effects were found. The results do not support the hypothesis that 100–300 μT MF modify biological responses to UV radiation by causing an overall change in oxidative reactions at cellular level.

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Breast cancer (BC) is the most common type of malignancy in female patients and radio-treatment is the conventional therapy even if a great number of studies reported that enhanced sensitivity to ionizing radiation as measured as chromosome effects is present in a significant proportion of cancer patients, including breast cancer ones. In this study we analysed whether peripheral blood lymphocytes from sporadic BC patients and healthy subjects showed a different sensitivity to ionizing radiation and whether cytogenetic radiosensitivity may serve as a breast cancer risk biomarker. To test this hypothesis, the in vitro radiation sensitivity was measured by using both G₀ and G₂ chromosome radiosensitivity assays, on 46 subjects (23 BC patients and 23 healthy subjects). Results show that cancer patients are more radiosensitive than healthy controls and that G₂ assay could be more appropriate to define the individual radiosensitivity if compared to G₀ assay.

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Boron Neutron Capture Therapy (BNCT) effectiveness depends on the preferential sequestration of boron in cancer cells relative to normal tissue cells. We present a novel strategy for sequestering boron using an adenovirus expressing the sodium iodide symporter (NIS). Human glioma grown subcutaneously in athymic mice and orthotopic rat brain tumors were transfected with NIS using a direct tumor injection of adenovirus. Boron bound as sodium tetrafluoroborate (NaBF₄) was administered systemically several days after transfection. Tumors were excised hours later and assessed for boron concentration using inductively coupled plasma atomic emission spectroscopy. In the human glioma transfected with NIS, boron concentration was more than 10 fold higher with 100 mg/kg of NaBF₄, compared to tumor not transfected. In the orthotopic tumor model, the presence of NIS conferred almost 4 times the boron concentration in rat tumors transfected with human virus compared with contralateral normal brain not transfected. We conclude that adenovirus expressing NIS has the potential to be used as a novel boron delivery agent and should be explored for future clinical applications.

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