Low-dose Γ-ray Irradiation Research Articles

Fabrication of PVDF membranes via γ-ray irradiation and investigation into their membrane formation mechanisms and water treatment properties

Polyvinylidene fluoride (PVDF) membrane is a cheap and commonly used separation material, but its inherent hydrophobicity results in significant membrane contamination, which limits its engineering application and further development. The hydrophilic PVDF-g/co-NVP membrane was prepared through low-dose γ-ray irradiation and using N-vinyl-2-pyrrolidinone (NVP) as a modifier in this study. The effects of γ-ray dose and incorporating NVP additive on the microstructure, hydrophilicity, anti-fouling capability, and mechanical capacity of membranes were discussed. Molecular dynamics (MD) simulation method was used to comprehensively investigate the impact of the NVP on the physicochemical characteristics of membrane structure. Under a specific NVP load, the water contact angle of the membrane decreases from 87.92° to 52.66°. The pure water permeance reaches 362.1 L/m2 h, and the porosity increases to 55.21 %±1.29 %, which indicates that γ-ray irradiation with NVP addition can effectively adjust the pore structure and hydrophilicity of the membrane. During anti-fouling test against bovine serum albumin (BSA), the modified membrane exhibits remarkable anti-fouling characteristics, including reduced BSA accumulation, enhanced flux recovery, lower irreversible contamination level, and extended cleaning cycle compared with the original pure PVDF membrane because of its higher hydrophilicity and positive zeta-potential value. This study elucidates the mechanism underlying γ-ray graft modification of PVDF membranes, thereby providing valuable insights into the development of efficient functional membrane materials.

Biological effects of low-dose γ-ray irradiation on chromosomes and DNA of Drosophila melanogaster.

While the damage to chromosomes and genes induced by high-dose radiation (HDR) has been well researched in many organisms, the effects of low-dose radiation (LDR), defined as a radiation dose of ≤100 mSv, are still being debated. Recent research has suggested that the biological effects of LDR differ from those observed in HDR. To detect the effect of LDR on genes, we selected a gene of Drosophila melanogaster, known as the multiple wing hair (mwh) gene. The hatched heterozygous larvae with genotype mwh/+ were irradiated by γ-rays of a 60Co source. After eclosion, the wing hairs of the heterozygous flies were observed. The area of only one or two mwh cells (small spot) and that of more than three mwh cells (large spot) were counted. The ratio of the two kinds of spots were compared between groups irradiated by different doses including a non-irradiated control group. For the small spot in females, the eruption frequency increased in the groups irradiated with 20–75 mGy, indicating hypersensitivity (HRS) to LDR, while in the groups irradiated with 200 and 300 mGy, the frequency decreased, indicating induced radioresistance (IRR), while in males, 50 and 100 mGy conferred HRS and 75 and 200 mGy conferred IRR. For the large spot in females, 75 mGy conferred HRS and 100–800 mGy conferred IRR. In conclusion, HRS and IRR to LDR was found in Drosophila wing cells by delimiting the dose of γ-rays finely, except in the male large spot.

Mice Exposed to Combined Chronic Low-Dose Irradiation and Modeled Microgravity Develop Long-Term Neurological Sequelae.

Spaceflight poses many challenges for humans. Ground-based analogs typically focus on single parameters of spaceflight and their associated acute effects. This study assesses the long-term transcriptional effects following single and combination spaceflight analog conditions using the mouse model: simulated microgravity via hindlimb unloading (HLU) and/or low-dose γ-ray irradiation (LDR) for 21 days, followed by 4 months of readaptation. Changes in gene expression and epigenetic modifications in brain samples during readaptation were analyzed by whole transcriptome shotgun sequencing (RNA-seq) and reduced representation bisulfite sequencing (RRBS). The results showed minimal gene expression and cytosine methylation alterations at 4 months readaptation within single treatment conditions of HLU or LDR. In contrast, following combined HLU+LDR, gene expression and promoter methylation analyses showed multiple altered pathways involved in neurogenesis and neuroplasticity, the regulation of neuropeptides, and cellular signaling. In brief, neurological readaptation following combined chronic LDR and HLU is a dynamic process that involves pathways that regulate neuronal function and structure and may lead to late onset neurological sequelae.

Developmental toxicity and apoptosis in zebrafish embryos induced by low-dose γ-ray irradiation.

In this paper, the developmental toxicity and apoptosis in zebrafish (Danio rerio) embryos induced by 0.01, 0.05, and 0.10-Gy γ-ray irradiation were investigated and verified by single cell gel electrophoresis, acridine orange staining, flow cytometry, transmission electron microscopy, digital gene expression sequencing, and Western blot analysis. DNA damage, deformity rates, and apoptosis of zebrafish embryos were found to increase significantly with the increase of irradiation dose, and survival and hatching rates significantly decreased when the irradiation dose exceeds 0.10 and 0.05Gy, respectively. Exposure to 0.10-Gy γ-ray irradiation resulted in the swelling of cell mitochondria of zebrafish embryos and changes in their intracellular vacuoles. mRNA and protein expression levels of Shh (sonic hedgehog 19KDa) and Smo (smoothened 86KDa) of Hh signaling pathway associated with the development of early embryos significantly increased with the increase of irradiation dose. Expression of the AKT (56KDa) and PiK3r3 (55KDa) genes, which are anti-apoptotic and involved with the PI3K/Akt signaling pathway, significantly decreased, while expression of the bada gene, which is pro-apoptotic, significantly increased. The results show that γ-ray irradiations of 0.01 and 0.05Gy can induce developmental toxicity and apoptosis in zebrafish embryos via Hh and PI3K/Akt signaling pathways, respectively.

Effects of γ-Ray Irradiation on the Radial Structure Heterogeneity in Polyacrylonitrile Fibers during Thermal Stabilization.

The radial structural heterogeneity of thermally-stabilized polyacrylonitrile (PAN) fiber is considered to be a limiting factor affecting the mechanical properties of the resulting carbon fibers. In this study, we demonstrate that a low-dose (60 kGy) γ-ray irradiation pretreatment can effectively mitigate the radial structural heterogeneity of PAN fibers after thermal stabilization. The characterization results indicate that low-dose γ-ray irradiation only affects the physical structure of PAN through decreasing its crystalline size and crystallinity, rather than inducing chemical cross-linking and/or intramolecular cyclization. It is proposed that an increased amorphous region in PAN fibers prompted by low-dose γ-ray irradiation can facilitate oxygen diffusion from skin to core during stabilization, which results in the increased structural homogeneity of stabilized PAN fibers.

Activation of denovo GSH synthesis pathway in mouse spleen after long term low-dose γ-ray irradiation

Glutathione (GSH) is an important cellular antioxidant and has a critical role in maintaining the balance of cellular redox. In this study, we investigated the GSH biosynthesis genes involved in the elevation of endogenous GSH levels using an irradiation system with an irradiation dose rate of 1.78 mGy/h, which was about 40,000 times less than the dose rates used in other studies. The results showed that GSH levels were significantly increased in the low-dose (0.02 and 0.2 Gy) irradiated group compared to those in the non-irradiated group, but enzymatic antioxidants such as superoxide dismutase and catalase were not induced at any doses tested. The elevation in GSH was accompanied by elevated expression of glutamate–cysteine ligase modifier subunit, but no changes were observed in the expression of glutamate–cysteine ligase catalytic subunit and thioredoxin in de novo GSH synthesis. In the case of genes involved in the GSH regeneration cycle, the expression of glutathione reductase was not changed after irradiation, whereas glutathione peroxidase was only increased in the 0.2 Gy irradiated group. Collectively, our results suggest that the de novo pathway, rather than the regeneration cycle, may be mainly switched on in response to stimulation with long-term low-dose radiation in the spleen.

Activation of Immune Functions by Low-dose, Whole-body Irradiation with .GAMMA.-Rays

We first examined whether the increase of glutathione level induced by low-dose γ-ray irradiation is involved in the appearance of enhanced natural killer (NK) activity and antibody-dependent cellular cytotoxicity (ADCC), leading to delayed tumor growth in Ehrlich solid tumor (EST) -bearing mice. NK activity in ICR mouse splenocytes was significantly increased from 4h to 6h after a single whole-body γ-ray irradiation at 0.5 Gy, and thereafter decreased almost to the zero-time level by 24h post-irradiation. ADCC was also increased significantly in a similar way. Reduced glutathione exogenously added to splenocytes obtained from normal mice enhanced both NK activity and ADCC in a dose-dependent manner. The inhibitory effect of the radiation on tumor growth was then examined in EST-bearing mice. Repeated low-dose irradiation (0.5 Gy, four times, before and within an early time after the inoculation) significantly delayed the tumor growth.Finally, the effect of single low-dose (0.5 Gy), whole-body γ-ray irradiation on immune balance was examined in order to elucidate the mechanism underlying the anti-tumor immunity. The percentage of B cells in blood lymphocytes was selectively decreased after the radiation, concomitantly with an increase in that of helper T cell population. The IFN-γ level in splenocyte culture prepared from EST-bearing mice was significantly increased 48h after the radiation, though the level of IL-4 was unchanged. IL-12 secretion from macrophages was also enhanced by the radiation. These results suggest that low-dose γ-rays induce Th1 polarization and enhance the activities of tumoricidal effector cells, leading to a delay of tumor growth.

Low dose gamma-rays activate immune functions via induction of glutathione and delay tumor growth.

We examined whether the increase of glutathione level induced by low dose gamma-ray irradiation is involved in the appearance of enhanced natural killer (NK) activity and antibody-dependent cellular cytotoxicity (ADCC), leading to delayed tumor growth in Ehrlich solid tumor-bearing mice. NK activity in ICR mouse splenocytes significantly increased from 4 h to 6 h after whole-body gamma-ray irradiation at 0.5 Gy, and thereafter decreased almost to the zero-time level by 24 h post-irradiation. ADCC also increased significantly in a similar way. Reduced glutathione exogenously added to splenocytes obtained from normal mice enhanced both NK activity and ADCC in a dose-dependent manner. Since immune functions were enhanced through the induction of cellular glutathione after low-dose irradiation, the inhibitory effect of the radiation on tumor growth was then examined in Ehrlich solid tumor-bearing mice. Tumor growth after inoculation was significantly delayed by the radiation. These results suggest that low-dose gamma-rays activate immune functions via an induction of glutathione, leading to a delay of tumor growth.

Increase of intracellular glutathione by low-dose gamma-ray irradiation is mediated by transcription factor AP-1 in RAW 264.7 cells.

The mechanism of the elevation of intracellular glutathione induced by low-dose gamma-rays was examined in RAW 264.7 cells. The expression of mRNA for gamma-glutamylcysteine synthetase (gamma-GCS) increased soon after gamma-ray (0.5 Gy) irradiation, and peaked between 3 h and 6 h post-irradiation. A dose of 0.25 to 0.5 Gy was optimum for induction of gamma-GCS mRNA expression at 3 h post-irradiation. The effect of inhibitors of activator protein-1 (AP-1) and nuclear factor kappaB (NF-kappaB) on the radiation-induced gamma-GCS gene expression was then examined. The induction of gamma-GCS mRNA expression was significantly suppressed when AP-1 DNA binding, but not NF-kappaB DNA binding, was inhibited. Finally, electrophoretic mobility shift assay showed that the low-dose radiation markedly increased the DNA binding of AP-1, but not NF-kappaB, soon after irradiation. These results suggest that the increase of glutathione levels in RAW 264.7 cells by low-dose gamma-ray irradiation is mediated by transcriptional regulation of the gamma-GCS gene, predominantly through the AP-1 binding site in its promoter.

Possible role of elevation of glutathione in acquisition of enhanced immune function of mouse splenocytes exposed to low-dose γ-rays

The relation between induction of an increased glutathione level and the enhanced immune functions, including proliferative response and natural killer (NK) activity, of mouse splenocytes by a low dose of γ-rays was investigated. Glutathione level in mouse splenocytes significantly increased 2 h after whole-body γ-ray irradiation at 0.5 Gy, peaked at 4 h and thereafter, decreased almost to the initial (0 h) level within 12 h after irradiation. A significant enhancement of Con A-induced proliferation was recognized in the splenocytes from the whole-body-irradiated animals obtained at post-irradiation between 2 and 6 h. NK activity was also enhanced at the same time periods after the irradiation. Addition of glutathione to splenocytes obtained from normal mice enhanced both Con A-induced proliferative response and NK activity in a dose-dependent manner. These enhancements were completely blocked by buthionine sulfoximine, a specific inhibitor of the de novo pathway of glutathione synthesis. These results suggest that induction of endogenous glutathione synthesis in splenocytes after low-dose γ-ray irradiation is partially responsible for the appearance of enhanced immune function.

Possible role of elevation of glutathione in the acquisition of enhanced proliferation of mouse splenocytes exposed to small-dose γ-rays

Purpose : To examine the relation between the induction of an increased glutathione level and the elevated proliferative response of mouse splenocytes by a small dose of γ-rays. Materials and methods : Male ICR strain mice, 7 weeks of age, were divided into irradiated and non-irradiated control groups. Irradiation was done with γ-rays from a 137 Cs source at a dose of 50cGy (1.11Gy/min). Glutathione content in the splenocytes was measured using a modified spectrophotometric technique. Concanavalin A (Con A)-induced proliferative response of the splenocytes after whole-body γ-ray irradiation was estimated from the 3 H-thymidine incorporation into the cells. Results : The glutathione level in mouse splenocytes increased 2h after whole-body γ-ray irradiation at 50 cGy, peaked at 4h and thereafter decreased almost to the zero-time level by 12-h postirradiation. A significant enhancement of Con A-induced proliferation was observed in the splenocytes obtained from the wholebody-irradiated animals between 2h and 6h post-irradiation. Glutathione exogenously added to splenocytes obtained from normal mice enhanced the Con A-induced proliferation of splenocytes in a dose-dependent manner. This enhancement was completely blocked by buthionine sulfoximine, a specific inhibitor of the de novo pathway of glutathione synthesis. Conclusions : The induction of endogenous glutathione immediately after low-dose γ-ray irradiation is at least partially responsible for the enhancement of immune function, and may throw light on the mechanisms of carcinostatic effects induced by low dose ionizing radiation.

Inhibitory effects of post low dose γ-ray irradiation on ferric-nitrilotriacetate-induced mice liver damage

We studied the effects of a single post whole-body low-dose irradiation (50 cGy of γ-ray) on mice with ferric nitrilotriacetate (Fe3+-NTA)-induced transient hepatopathy. As a result, low-dose irradiation accelerated the rate of recovery. Based on the changes in glutamic oxaloacetic transaminase (GOT) activities, glutamic pyruvic transaminase (GPT) activities and lipid peroxide levels, it was shown that hepatopathy was improved by low-dose irradiation 3 h after Fe3+-NTA administration. This may be because of the enhancement of antioxidant agents such as total glutathione (GSH + GSSG), glutathione peroxidase (GPX), glutathione reductase (GR) and γ-glutamylcysteine synthetase (γ-GCS) by low-dose irradiation. These findings suggest that low-dose irradiation relieved functional disorders at least in the livers of mice with active oxygen species related diseases.

Low-dose γ-ray irradiation reduces oxidative damage induced by CCl 4 in mouse liver

We examined the effects of irradiation (50 cGy of γ-ray) reducing the oxidative damage in carbon tetrachloride (CCl 4)-hepatopathy mice. We made pathological examinations and analyzed transaminase activity (glutamic oxaloacetic transaminase and glutamic pyruvic transaminase), lipid peroxide level and the activities of endogenous antioxidants in the mouse. The irradiation was found to accelerate the recovery. Based on pathological examination as well as changes in each transaminase activity and lipid peroxide levels, it was shown that hepatopathy improved 3 d after the irradiation. The activities of glutathione reductase and glutathione peroxidase rapidly elevated after irradiation, and the total glutathione content gradually increased in the irradiation group. Both activities of γ-glutamylcysteine synthetase and catalase were higher than normal at all times after the irradiation and gradually increased. In addition, the γ-glutamylcysteine synthetase activity changed in a similar fashion to the total glutathione content. However, superoxide dismutase activity in both groups decreased and that of the irradiation group was significantly lower than that of the sham-irradiation group. These findings suggest that low-dose radiation relieved functional disorder at least in the liver of mice with active oxygen diseases.

Elevation of antioxidant potency in the brain of mice by low-dose γ-ray irradiation and its effect on 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced brain damage

The elevation of endogenous thiol-related antioxidants and free radical scavenging enzymes in the brain of C57BL/6 female mice after low-dose γ-ray irradiation and its inhibitory effect on 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced brain damage were investigated. The brain level of the reduced form of glutathione (GSH) increased soon after irradiation with 50 cGy of γ-rays, reached a maximum at 3 h post-treatment, and remained elevated until 12 h. Thioredoxin (TRX) was also transiently increased after irradiation. The activities of free radical scavenging enzymes, including Cu/Zn-superoxide dismutase, catalase and glutathione peroxidase, were significantly induced after irradiation as well. Cerebral malondialdehyde was remarkably elevated by MPTP treatment, and this elevation was suppressed by pre- irradiation (50 cGy). The contents of GSH and TRX were significantly decreased by MPTP treatment in comparison with those of the control group. These reductions both seemed to be attenuated by pre-irradiation with γ-rays. These results suggest that low-dose γ-ray irradiation induces endogenous antioxidative potency in the brain of mice and might be effective for the prevention and/or therapy of various reactive oxygen species-related neurodegenerative disorders, such as Parkinson’s disease and Alzheimer’s disease.

Localization of glutathione and induction of glutathione synthesis-related proteins in mouse brain by low doses of γ-rays

First, we determined the cerebral localization of reduced glutathione (GSH) in normal mice by means of autoradiography using 99mTc - meso-hexamethyl propylene oxime. A highly specific localization of GSH in the cerebellum and hippocampus was observed. Secondly, we measured the elevation of GSH level in the brain after low-dose γ-irradiation. The cerebral GSH levels increased soon after irradiation with 50 cGy of γ-rays, reaching a maximum at 3 h post-treatment, then remaining significantly higher than that of the non-irradiated control until 12 h and returning to the control level by 24 h. Thirdly, we examined the induction of the activities and the mRNAs of proteins involved in the synthesis and regeneration of GSH in the brain of mice subjected to low-dose γ-ray irradiation. The level of mRNA for γ-glutamylcysteine synthetase was significantly increased at 0.5 h, and remained high until 2 h post-irradiation (50 cGy). The level was transiently lowered to the non-irradiated control level at 3 h and slightly increased again after 6 h post-irradiation. γ-Glutamylcysteine synthetase activity was significantly increased 3 h after irradiation, and remained high up to 24 h post-irradiation. As for glutathione reductase, the mRNA level was increased at 0.5 h, and peaked strongly at 2 h, while the enzyme activity was significantly increased at 6 h after irradiation, and continued to increase up to 24 h. The level of mRNA for thioredoxin, which contributes to GSH biosynthesis by supplying cysteine to the de novo pathway, peaked between 0.5 h and 2 h post-irradiation, and rapidly declined thereafter. The content of thioredoxin showed a transient decrease immediately after irradiation, but was then remarkably elevated, reaching a maximum at 3 h, and thereafter declining sharply. These results indicate that the increase in endogenous GSH in mouse brain soon after low-dose γ-ray irradiation is a consequence of the induction of GSH synthesis-related proteins and occurs via both the de novo synthesis and the regeneration pathways.

Induction of mRNAs for glutathione synthesis-related proteins in mouse liver by low doses of γ-rays

We examined the elevation of the reduced form of glutathione (GSH) level and the induction of mRNAs for proteins involved in the synthesis and regeneration of GSH in the liver of mice after low-dose γ-ray irradiation. The liver GSH level increased soon after irradiation with 50 cGy of γ-rays, reached a maximum at around 12 h post-treatment. The mRNA of γ-glutamylcysteine synthetase (γ-GCS), the rate-limiting enzyme for de novo synthesis for GSH, showed a small increase that peaked at 6 h after γ-ray irradiation at a dose of 50 cGy. Only a small increase in γ-GCS activity was observed throughout the 24-h post-irradiation period. In the case of glutathione reductase (GR), which is involved in the regeneration of GSH from the oxidized form (GSSG), the mRNA level peaked strongly at 1 h, while the activity peaked at twice the control level 12 h after irradiation. The level of mRNA for thioredoxin (TRX), which contributes to GSH biosynthesis by supplying cysteine to the de novo pathway, peaked at 1 h and declined thereafter, while the activity peaked at 3 h and then declined sharply. These results indicate that the increase in endogenous GSH immediately following low-dose γ-ray irradiation is predominantly due to operation of the regeneration cycle and not de novo synthesis. We also examined the dependence of mRNA induction on the γ-ray dose.

Protein biosynthesis in low dose ionizing radiation-adapted human melanoma cells.

Adaptive responses induced by low dose gamma-ray irradiation in human melanoma cells were examined using a clonogenic assay. Survival fractions were significantly increased in cells irradiated with low dose gamma-rays then 4 hrs later with high dose gamma-ray as compared to cells irradiated only with high dose gamma-rays. When low dose irradiation was given 20 hrs prior to high dose irradiation, however, no adaptive response was induced. Changes in protein biosynthesis in human melanoma cells were observed under the same conditions. Significant changes in protein biosynthesis occurred in the nuclear and membrane proteins of cells first irradiated with a low dose then a high dose of gamma-rays after 4 hrs. No such changes were found in cells irradiated with low dose gamma-rays 20 hrs prior to high dose irradiation, consistent with the results of the clonogenic assay. Our findings suggest that prior treatment with low dose gamma-rays induces an adaptive response that has a significant effect on the induction of the nuclear and membrane protein biosynthesis caused by high dose gamma-ray irradiation of human melanoma cells.

Adaptive response induced by low dose ionizing radiation in human cervical carcinoma cells

Adaptive response induced by low dose γ-ray irradiation in human cervical carcinoma cells was examined. Cells were exposured to low dose of γ-ray (1 cGy) followed by high doses of γ-ray irradiation (0, 1, 2, 3, 5, 7 and 9 Gy for clonogenic assay or 1.5 Gy for micronucleus assay) with various time intervals. Survival fractions of cells in both low dose-irradiated and unirradiated groups were analyzed by clonogenic assay. Survival fractions of low dose-irradiated cells was higher than that of control group irradiated only with high dose γ-ray. The increase in cell survival was maximum when low and high dose irradiation time interval was 4 hr. Frequencies of micronuclei which is an indicative of chromosome aberration were also enumerated in both low dose-irradiated and unirradiated groups. In consistent with the result obtained from survival fractions analyzed by clonogenic assay, maximum reduction in frequencies of micronuclei was observed when low dose radiation was given 4 hr prior to high dose irradiation. These results demonstrate that low dose γ-ray irradiation induced adaptive response to subsequent high dose γ-ray irradiation in human cervical carcinoma cells. Our data suggest that one of the possible mechanisms of adaptive response induced by low dose radiation is the increase in repair of DNA double strand breaks in low dose radiation-adapted cells.