Differential expression of thymic DNA repair genes in low-dose-rate irradiated AKR/J mice.

We previously determined that AKR/J mice housed in a low-dose-rate (LDR) ((137)Cs, 0.7 mGy/h, 2.1 Gy) γ-irradiation facility developed less spontaneous thymic lymphoma and survived longer than those receiving sham or high-dose-rate (HDR) ((137)Cs, 0.8 Gy/min, 4.5 Gy) radiation. Interestingly, histopathological analysis showed a mild lymphomagenesis in the thymus of LDR-irradiated mice. Therefore, in this study, we investigated whether LDR irradiation could trigger the expression of thymic genes involved in the DNA repair process of AKR/J mice. The enrichment analysis of Gene Ontology terms and Kyoto Encyclopedia of Genes and Genomes pathways showed immune response, nucleosome organization, and the peroxisome proliferator-activated receptors signaling pathway in LDR-irradiated mice. Our microarray analysis and quantitative polymerase chain reaction data demonstrated that mRNA levels of Lig4 and RRM2 were specifically elevated in AKR/J mice at 130 days after the start of LDR irradiation. Furthermore, transcriptional levels of H2AX and ATM, proteins known to recruit DNA repair factors, were also shown to be upregulated. These data suggest that LDR irradiation could trigger specific induction of DNA repair-associated genes in an attempt to repair damaged DNA during tumor progression, which in turn contributed to the decreased incidence of lymphoma and increased survival. Overall, we identified specific DNA repair genes in LDR-irradiated AKR/J mice.

Ameliorating effect of new constituents from the hooks of Uncaria rhynchophylla on scopolamine-induced memory impairment.

AIM: To study the chemical constituents and their anti-amnesic effect from the hooks of Uncaria rhynchophylla. METHODS: The isolation of compounds was performed by chromatographic techniques and their structures were identified on the basis of spectral analysis. Their ameliorating effects on scopolamine-induced memory impairment in vivo using a Morris water-maze task and passive avoidance task system were evaluated. RESULTS: Activity-guided fractionation of the total extracts resulted in the isolation of four constituents, trans-anethole (1), p-anisaldehyde (2), estragole (3), and 3-oxo-olean-12-en-28-oic acid (4), which were found for the first time from this plant. CONCLUSION: Compound 1 exhibited a better memory enhancing effect than tacrine, a positive agent, at the same dose in the passive avoidance test and a similar property in the water-maze test, and its action may be mediated, in part, by the acetylcholine enhancing cholinergic nervous system.

Identification of radiation-sensitive expressed genes in the ICR and AKR/J mouse thymus.

We have investigated radiation-sensitive expressed genes (EGs), their signal pathways, and the effects of ionizing radiation in the thymus of ICR and AKR/J mice. Whole-body and relative thymus weights were taken and microarray analyses were done on the thymuses of high-dose-rate (HDR, (137) Cs, 0.8 Gy/min, a single dose of 4.5 Gy) and low-dose-rate (LDR, (137) Cs, 0.7 mGy/h, a cumulative dose of 1.7 Gy) irradiated ICR and AKR/J mice. Gene expression patterns were validated by quantitative polymerase chain reaction (qPCR). The effect of ionizing radiation on thymus cell apoptosis was measured terminal deoxynucleotidyl-transferase-mediated dUTP-end labeling (TUNEL). LDR-irradiation increased the mean whole-body weight, but decreased the relative thymus weight of AKR/J mice. Radiation-sensitive EGs were found by comparing HDR- and LDR-irradiated ICR and AKR/J mice. qPCR analysis showed that 12 EGs had dose and dose-rate dependent expression patterns. Gene-network analysis indicated that Ighg, Igh-VJ558, Defb6, Reg3g, and Saa2 may be involved in the immune response, leukocyte migration, and apoptosis. Our data suggest that expression of the HDR (Glut1, Glut4, and PKLR) and LDR radiation-response genes (Ighg and Igh-VJ558) can be dose or dose-rate dependent. There was an increased number of apoptotic cells in HDR-irradiated ICR mice and LDR-irradiated AKR/J mice. Thus, changes of the mean whole-body weight and relative thymus weight, EGs, signal pathways, and the effects of ionizing radiation on the thymus of ICR and AKR/J mice are described.

Augmentation of natural cytotoxicity by chronic low-dose ionizing radiation in murine natural killer cells primed by IL-2.

The possible beneficial effects of chronic low-dose irradiation (LDR) and its mechanism of action in a variety of pathophysiological processes such as cancer are a subject of intense investigation. While animal studies involving long-term exposure to LDR have yielded encouraging results, the influence of LDR at the cellular level has been less well defined. We reasoned that since natural killer (NK) cells constitute an early responder to exogenous stress, NK cells may reveal sentinel alterations in function upon exposure to LDR. When purified NK cells received LDR at 4.2 mGy/h for a total of 0.2 Gy in vitro, no significant difference in cell viability was observed. Likewise, no functional changes were detected in LDR-exposed NK cells, demonstrating that LDR alone was insufficient to generate changes at the cellular level. Nonetheless, significant augmentation of cytotoxic, but not proliferative, function was detected when NK cells were stimulated with low-dose IL-2 prior to irradiation. This enhancement of NK cytotoxicity was not due to alterations in NK-activating receptors, NK1.1, NKG2D, CD69 and 2B4, or changes in the rate of early or late apoptosis. Therefore, LDR, in the presence of suboptimal cytokine levels, can facilitate anti-tumor cytotoxicity of NK cells without influencing cellular proliferation or apoptosis. Whether these results translate to in vivo consequences remains to be seen; however, our data provide initial evidence that exposure to LDR can lead to subtle immune-enhancing effects on NK cells and may explain, in part, the functional basis underlying, diverse beneficial effects seen in the animals chronically exposed to LDR.

Sperm abnormalities in high- and low-dose-rate Gamma-irradiated Korean dark-striped field mice, Apodemus agrarius coreae.

To evaluate the in vivo effects of low-dose-rate (0.7 mGy h(-1)) gamma radiation, abnormal shapes of sperm in the caudal epidydimus of Apodemus agrarius coreae (A. a. coreae) were used. The six categories of abnormal forms (amorphous heads, blunt hooks, excessive hooks, two heads and tails, folded tails and short tails) of sperm were observed eight days after gamma irradiation (0, 0.5, 1 and 2 Gy) with a high dose rate (0.8 Gy min(-1)) and a low dose rate. The frequency of total abnormal sperm gradually increased starting from 0.5 Gy after high-dose-rate radiation. Blunt hooks and short tails shaped sperm, in particular, were gradually increasing in the high-dose-rate irradiated mice. Dose rate reduction effects for the frequency of abnormal sperms in low-dose-rate irradiated mice to high-dose-rate irradiated mice were 1 at 0.5 Gy, 0.7 at 1 Gy and 0.5 at 2 Gy. Our results indicate that low-dose-rate radiation is not detrimental to spermatogenic cells.

Differential expression of immune-associated cancer regulatory genes in low- versus high-dose-rate irradiated AKR/J mice.

AKR/J mice carrying leukemia viral inserts develop thymic lymphoma. Recently, we demonstrated that the incidence of thymic lymphoma was decreased when these mice were raised in a low-dose-rate γ-irradiation facility. In contrast, mice irradiated at a high-dose rate developed severe thymic lymphoma and died much earlier. To understand the genetic changes occurred by low- versus high-dose-rate γ-irradiation whole genome microarray was performed. Both groups of mice demonstrated up-regulation of Ifng, Igbp1, and IL7 in their thymuses, however, mice exposed to high-dose-rate γ-irradiation exhibited marked down-regulation of Sp3, Il15, Traf6, IL2ra, Pik3r1, and Hells. In contrast, low-dose-rate irradiated mice demonstrated up-regulation of Il15 and Jag2. These gene expression profiles imply the impaired immune signaling pathways by high-dose-rate γ-irradiation while the facilitation of anti-tumor immune responses by low-dose-rate γ-irradiation. Therefore, our data delineate common and distinct immune-associated pathways downstream of low- versus high-dose-rate irradiation in the process of cancer progression in AKR/J mice.

Life span and thymic lymphoma incidence in high- and low-dose-rate irradiated AKR/J mice and commonly expressed genes.

To evaluate the effect of low-dose-rate radiation on cancer incidence, we housed AKR/J mice in a long-term low-dose-rate irradiation facility ((137)Cs, 0.07 cGy/h). We compared the thymic lymphoma incidence and life span with those of mice irradiated at a high dose rate ((137)Cs, 0.8 Gy/min, total dose of 4.5 Gy) and nonirradiated mice. The average life span of the low-dose-rate irradiated mice (243 days) was longer than those of the high-dose-rate irradiated mice (208 days) and nonirradiated mice (230 days) (P = 0.02). The incidence of thymic lymphoma in low-dose-rate irradiated mice was lower than that in nonirradiated mice and high-dose-rate irradiated mice by 10 and 20%, respectively (P < 0.01). Normal-sized thymuses were collected 130 days after irradiation, and whole genome microarray analysis was performed. A total of 17,625 genes were assessed. Up- and down-regulated genes in low-dose-rate irradiated mice were 1.7 and 9 times less frequent than in high-dose-rate irradiated mice. We profiled expressed genes associated with carcinogenesis pathways (DNA repair, DNA damage signaling pathway, cell cycle, cancer pathway finder, p53 signaling pathway, apoptosis and T-cell and B-cell activation). Apoptosis- (Cd5l, Fcgr3 and Pycard) and immune- (Pycard, Lilrb3, Igh-6, Fcgr2b and MGC60843) related genes were commonly activated in both high- and low-dose-rate irradiated mice. The results suggest that carcinogenic cells have been removed by activated apoptosis and immune mechanisms, contributing to decreased thymic lymphoma and elongated life span. Functional studies for expressed genes associated with thymic lymphoma incidence in low-dose-rate exposed mice are currently under way.

Alteration of cytokine profiles in mice exposed to chronic low-dose ionizing radiation.

While a high-dose of ionizing radiation is generally harmful and causes damage to living organisms, a low-dose of radiation has been shown to be beneficial in a variety of animal models. To understand the basis for the effect of low-dose radiation in vivo, we examined the cellular and immunological changes evoked in mice exposed to low-dose radiation at very low (0.7mGy/h) and low (3.95mGy/h) dose rate for the total dose of 0.2 and 2Gy, respectively. Mice exposed to low-dose radiation, either at very low- or low-dose rate, demonstrated normal range of body weight and complete blood counts. Likewise, the number and percentage of peripheral lymphocyte populations, CD4(+) T, CD8(+) T, B, or NK cells, stayed unchanged following irradiation. Nonetheless, the sera from these mice exhibited elevated levels of IL-3, IL-4, leptin, MCP-1, MCP-5, MIP-1alpha, thrombopoietin, and VEGF along with slight reduction of IL-12p70, IL-13, IL-17, and IFN-gamma. This pattern of cytokine release suggests the stimulation of innate immunity facilitating myeloid differentiation and activation while suppressing pro-inflammatory responses and promoting differentiation of naïve T cells into T-helper 2, not T-helper 1, types. Collectively, our data highlight the subtle changes of cytokine milieu by chronic low-dose gamma-radiation, which may be associated with the functional benefits observed in various experimental models.

Relative morphological abnormalities of sperm in the caudal epididymis of high- and low-dose-rate gamma-irradiated ICR mice.

This study evaluated the effects of low dose radiation on spermatogenic cells using the morphological characteristics of sperm in the caudal epididymis of ICR mice. In this study, six abnormal sperm shapes (amorphous heads, blunt hooks, excessive hooks, two heads and tails, folded tails and short tails) were observed at eight days after gamma-irradiation ((137)Cs, 0, 0.2, 0.5, 1, 2 or 4 Gy) with both a high-dose-rate (0.8 Gy/min) and a low-dose-rate (0.7 mGy/hr). Fewer abnormal forms of sperm were observed in low-dose-rate irradiated mice than in mice that received a high-dose-rate irradiation (P = 0.002). The ratio of the dose rate effect among low-dose-rate irradiated mice to high-dose-rate irradiated mice was approximately 0.6. In addition, sperm with blunt hooks and two heads and tails significantly increased in number after irradiation, potentially providing an endpoint marker for estimating the effects of radiation. This study suggests that low-dose-rate (0.7 mGy/hr) radiation does not damage stem spermatogonia and probably stimulates repair in damaged spermatogonial stem cells in male mice.