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Purpose: Baicalein (an anti-ferroptosis drug) was recently reported to synergistically improve the survival rate of mice following a high dose of total body irradiation with anti-apoptosis and anti-necroptosis drugs. At the same time, our group has demonstrated that ferrostatin-1, a ferroptosis inhibitor, improves the survival rate of a mouse model of hematopoietic acute radiation syndrome to 60% for 150 days ( p  < .001). These phenomena suggest that ferroptosis inhibition can mitigate radiation damage. In this study, we continued to study the mechanisms by which ferrostatin-1 alleviated radiation-induced ferroptosis and subsequent hematopoietic acute radiation syndrome., Materials and Methods: Male ICR mice (8-10 weeks old) were exposed to doses of 0, 8, or 10 Gy irradiated from a 137 Cs source. Ferrostatin-1 was intraperitoneally injected into mice 72 h post-irradiation. Bone marrow mononuclear cells (BMMCs) and peripheral blood cells were counted. The changes in iron-related parameters, lipid metabolic enzymes, lipid peroxidation repair molecules (glutathione peroxidase 4, glutathione, and coenzyme Q10), and inflammatory factors (TNF-α, IL-6, and IL-1β) were evaluated using biochemical or antibody techniques., Results: Ferrostatin-1 increased the number of red and white blood cells, lymphocytes, and monocytes in the peripheral blood after total body irradiation in mice by mitigating the ferroptosis of BMMCs. Total body irradiation induced ferroptosis in BMMCs by increasing the iron and lipid peroxidation levels and depleting the acyl-CoA synthetase long-chain family member 4 (ASCL4), lipoxygenase 15, glutathione peroxidase 4, and glutathione levels. Ferroptotic BMMCs did not release TNF-α, IL-6, or IL-1β at the early stage of radiation exposure. Ferrostatin-1 mitigated the lipid peroxidation of radiation-induced ferroptosis by attenuating increases in levels of hemosiderin and liable iron pool and decreases in levels of ASCL4 and glutathione peroxidase 4., Conclusions: The onset of total body irradiation-induced ferroptosis in BMMCs involved changes in iron, lipid metabolic enzymes, and anti-lipid peroxidation molecules. Ferrostatin-1 could be a potential radiation mitigation agent by acting on these targets.

Background: UV radiation and its impact on living organisms became an essential concern over the past three decades and will be essential in the years to come. So, the present investigation was devoted to examining the impact of artificial UV-B radiation on the accumulation of amino acids and MDA contents as well as some antioxidant enzymes activities in three freshwater cyanobacterial species; Planktothrix cryptovaginata, Nostoc carneum and Microcystis aeruginosa , one freshwater green alga; Scenedesmus acutus and one marine cyanobacterium; Microcystis ., Methods: The algal cultures were exposed directly to artificial UV-B radiation for 1, 3, 5, and 7 hours and amino acids, MDA contents, and the antioxidant enzyme activities; CAT, POD, APX, and SOD were analyzed., Results: The data obtained indicated that alteration in MDA and antioxidant enzymes by UV stress depends on the algal species and the exposure time. The treatment of the investigated algae with different periods of UV-B exposure stimulated the biosynthesis of some individual amino acids and inhibited the accumulation of some others. In some cases, exposure to UV-B was accompanied by the disappearance of some amino acids. In addition, UV-B exposure for 3 hours stimulated the accumulation of total amino acids in M. aeruginosa and S. acutus , while 7 hours of UV-B enhanced the biosynthesis of total amino acids in M. aeruginosa only from the investigated algae., Conclusion: Exposure of some cyanobacteria and green algae to UV-B radiation stimulated the biosynthesis of some individual amino acids and inhibited the accumulation or accompanied by the disappearance of some others. However, the alteration in MDA and antioxidant enzymes by UV stress depends on the algal species and the exposure time.

Purpose: Light intensity is one of the important environmental factors that have strong influence on larviculture. Culture of fish at high intensity of light generates harmful radicals in the body that can compromise their health and production. The present study aims to evaluate the effect of various light intensities on the physiology of rohu Labeo rohita fry. Materials and Methods: Rohu fry (13.56 ± 0.4 mg) were exposed at five different light intensities: 0.17 ± 0.005 (Lc, control), 1.45 ± 0.23 (L1), 2.69 ± 0.47 (L2), 3.93 ± 0.72 (L3) and 5.06 ± 0.95 Wm -2 (L4). After 90 days of culture, rohu were harvested. Results and Conclusions: A 2-5% mortality of rohu was recorded in L3 and L4 treatments. The average weight and specific growth rate were significantly ( p  < .05) higher in Lc treatment compared to others. The light intensity and swimming activity of rohu showed direct relationship, whereas, light intensity showed inverse relation with nitric oxide synthase and reduced glutathione levels. Significantly ( p  < .05) higher glutathione S-transferase and glutathione peroxidase activies were found in rohu exposed at L4 treatment. Higher light intensities resulted in oxidative stress in the muscles of rohu. Thiobarbituric acid reactive substances, carbonyl protein and heat shock protein 70 were significantly ( p  < .05) higher in rohu exposed at L4 compared to other treatments. Exposure of rohu fry to intense light resulted into physiological stress and immunosupression.

Purpose: The present study focused on the role of primary metabolites, antioxidant activities in black gram ( Vigna mungo L. Hepper) variety Vamban-4 under the impact of gamma irradiation. Materials and methods: The black gram seeds were irradiated at a different dose of gamma rays viz., 200, 400, 600, 800, 1000 and 1200 Gray (Gy), and analyzed the changes of biochemical and antioxidant contents. Results: Biochemical contents increased with increasing doses up to 800 Gy and reduced at higher dose viz.,1000 and 1200 Gy of gamma-ray treatment, while malondialdehyde content increased at higher dose 1200 Gy compared to control, this indicates that the formation of more lipid peroxidation at higher radiation level. Gamma irradiation enhanced the activity of antioxidant enzymes such as catalase, peroxidase and superoxide dismutase with increasing doses rather than control plants. These studies depict that the effect of gamma irradiation on seeds increased the free radicals compared to control, which emphasize by the Electron spin resonance spectroscopy (ESR). Gamma irradiated seed samples contain more fractions of protein, lipids, amino acid and polysaccharides compared to control plants observed from the Fourier infrared transform spectroscopy (FTIR). Conclusion: The present result concludes that cellular metabolites and the antioxidant enzymes increased and these alterations of metabolism due to the effect of gamma rays which induced free radical production in seeds and plants grown under irradiation stress. These results confirm that the exposure of gamma irradiation produced free radicals as well as counteract by antioxidant enzymes and in addition to that accumulation and alteration of primary metabolites.

Purpose: The regular low dose occupational exposure to ionizing radiation may induce deleterious health effects, which may be of particular interest to medical radiation workers who daily handle X-ray machines. Human peripheral blood lymphocytes are able to retain the signature of radiation-induced DNA damage, therefore, the present study was undertaken to investigate the DNA damage and antioxidants status in hospital workers occupationally exposed to low doses of X-rays. Materials and methods: The peripheral blood lymphocytes of the occupationally exposed and control groups matched for age, gender, tobacco usage, and alcohol consumption were cultured and micronuclei frequency was determined. Activities of antioxidant enzymes and lipid peroxidation were also estimated in their plasma. Results: The micronuclei frequency in the occupationally exposed group ( n  = 33), increased significantly ( p  < .0001) followed by reduced glutathione-s-transferase ( p  < .01) and catalase ( p  < .001) activities, and increased lipid peroxidation ( p  < .05) when compared to the control group ( n  = 33). Occupational exposure resulted in an effective dose ranging between 3.14 to 144.5 mSv (40.88 ± 39.86mSv) depending on the employment duration of 3-29 years (10.33 ± 7.05 years). A correlation between the micronuclei frequency ( p  < .05) and catalase activity ( p  < .05) existed in the occupationally exposed individuals depending on the smoking habit, age, duration of employment, cumulative exposure dose and number of patients handled per day. Conclusions: We have observed that protracted low dose exposure to ionizing radiation is an inevitable occupational hazard leading to persistence of oxidative stress and increased genomic instability in the radiological technicians depending on the time spent with X-rays, cumulative dose received and the number of patients handled daily raising the risk of cancer development.

Purpose: To investigate the effect of whole-body low-dose radiation (10 cGy of 6 MV x-rays) given prior to a challenging dose of 2 Gy x-rays on adaptive response in the liver of rats Material and methods: Rats were either irradiated with 10 cGy, 2 Gy or 10 cGy 24 h prior to 2 Gy (10 cGy-2 Gy irradiated). Liver samples were analyzed for apoptosis, caspase-3, Bcl-2, by flow cytometry. DNA damage was determined by alkaline comet assay. The protein damage, lipid damage, and expression of endogenous antioxidants, Il-1β, Il-6, and TNF-α were determined by ELISA., Results: An increase in apoptosis paralleled with increase in DNA damage and expression of Il-1β, Il-6, and TNF-α, protein oxidation, and lipid peroxidation were detected in all irradiated groups. The irradiated rats with 10 cGy-2 Gy and the 2 Gy-irradiated showed the highest effect. The increase in oxidative damage of protein and lipid was accompanied with depletion of endogenous antioxidants. In all cases, the10 cGy-2 Gy irradiated group showed similar or higher effect than the 2 Gy-irradiated group., Conclusions: Under the present experimental conditions, There is no evidence that 10 cGy-irradiation protected the liver cells against a subsequent higher dose. The significant effect of 10 cGy-irradiation emphasizes the low-dose radiation-associated risk.

Purpose: Radiation-induced lung injuries (RILI), namely radiation pneumonitis and/or fibrosis, are dose-limiting outcomes following treatment for thoracic cancers. As part of a search for mitigation targets, we sought to determine if persistent DNA damage is a characteristic of this progressive injury., Methods: C57BL/6J female mice were sacrificed at 24 h, 1, 4, 12, 16, 24 and 32 weeks following a single dose of 12.5 Gy thorax only gamma radiation; their lungs were compared to age-matched unirradiated animals. Tissues were examined for DNA double-strand breaks (DSBs) (γ-H2A.X and p53bp1), cellular senescence (senescence-associated beta-galactosidase and p21) and oxidative stress (malondialdehyde)., Results: Data revealed consistently higher numbers of DSBs compared to age-matched controls, with increases in γ-H2A.X positivity beyond 24 h post-exposure, particularly during the pathological phases, suggesting periods of recurrent DNA damage. Additional intermittent increases in both cellular senescence and oxidative stress also appeared to coincide with pneumonitis and fibrosis., Conclusions: These novel, long-term data indicate (a) increased and persistent levels of DSBs, oxidative stress and cellular senescence may serve as bioindicators of RILI, and (b) prevention of genotoxicity, via mitigation of free radical production, continues to be a potential strategy for the prevention of pulmonary radiation injury.

Purpose: There is a great concern regarding the possible adverse effects of electromagnetic radiation (EMR). This study investigated the effects of EMR induced by Wi-Fi (2.45 GHz) on insulin secretion and antioxidant redox systems in the rat pancreas., Materials and Methods: Adult male Sprague-Dawley rats in the weight range of 230-260 g were divided into control, sham, Wi-Fi exposed groups. After long-term exposure (4 h/day for 45 days) to Wi-Fi EMR, plasma levels of glucose and insulin during intraperitoneal glucose tolerance test were measured. Islet insulin secretion and content, lipid peroxidation, and antioxidant status in pancreas of rats were determined., Results: Our data showed that the weight gain in the WI-FI exposed group was significantly lower than the control group (p < .05). Wi-Fi (2.45 GHz)-exposed group showed hyperglycemia. Plasma insulin level and glucose-stimulated insulin secretion from pancreatic islet were significantly reduced in the Wi-Fi-exposed group. EMR emitted from Wi-Fi caused a significant increase in lipid peroxidation and a significant decrease in GSH level, SOD, and GPx activities of the pancreas., Conclusions: These data showed that EMR of Wi-Fi leads to hyperglycemia, increased oxidative stress, and impaired insulin secretion in the rat pancreatic islets.

Purpose: The current study was undertaken to evaluate radioprotective effects of selenium (Se) nanoparticles in irradiation-induced nephropathy of mice model compared to sodium selenite., Materials and Methods: Forty-five mice were divided into three major groups including control, Se nanoparticle, and sodium selenite. Each major group was further subdivided into three more groups receiving various doses of 0, 2, and 8 Gy gamma irradiation. Both of the supplements were administered intraperitoneally with the dose of 0.1 mg/kg for 14 consecutive days. At the end of each week, the animals were exposed to gamma radiation and 48 h after the last exposure, the animals were humanely euthanized, then blood and renal tissue samples were taken. Serum creatinine, urea, cystatin C, and beta-2-microglobulin levels as well as activities of renal antioxidant enzymes, superoxide dismutase, glutathione peroxidase and catalase, also malondialdehyde level, total antioxidant capacity, renal tissue Se content, and histopathological features were assessed., Results: The results showed that both of the supplements could normalize aforementioned indices. However, selenium nanoparticles (Se-NPs) were more effective than sodium selenite., Conclusions: Conclusively, Se-NPs as an emerging potent antioxidant agent can protect against irradiation-induced nephropathy.

Purpose: Environmental changes generate free radicals and reactive oxygen species (ROS) resulting in abiotic stress in plants. This causes alterations in germination, morphology, growth and development ultimately leading to yield loss. Gamma irradiation was used to experimentally induce oxidative damage in an important pulse crop Vigna radiata (L.) Wilczek or mung bean. Our research was aimed towards augmentation of oxidative stress tolerance through treatment with a group of aliphatic amines known as polyamines., Materials and Methods: We used sub-lethal doses of gamma irradiation to generate oxidative damage which was evaluated using Nitro blue tetrazolium (NBT) staining, total antioxidant activity, 1, 1-Diphenyl-2-picryl hydrazyl (DPPH) radical scavenging assay, proline content and lipid peroxidation. Changes in internal free polyamines and messenger ribonucleic acid (mRNA) expression of key rate-limiting S-adenosylmethionine decarboxylase (SAMDC) enzyme in polyamine biosynthetic pathway was studied using real-time polymerase chain reaction (PCR)., Results: We observed increased oxidative damage with higher irradiation dose which was partially alleviated by putrescine treatment. Internal levels of putrescine and spermidine increased with 1 mM (50 and 100 Gy) and 2 mM putrescine treatment. Expression of SAMDC also increased with putrescine treatment., Conclusion: This study shows that treatment with putrescine can partially alleviate oxidative damage caused by gamma rays.

Purpose: To compare the effects of gamma-irradiation on biochemical responses and growth, six-week-old Brachypodium plants were chronically exposed to gamma-irradiation for 30 days at various dosages., Materials and Methods: Growth surveys of Brachypodium plants in response to different dosages of gamma-irradiation were conducted to compare physiological changes between irradiated and non-irradiated plants. Photosynthetic pigments, soluble sugar content, activities of antioxidant enzymes, and malonaldehyde (MDA) induced by reactive oxygen species (ROS) production were also measured., Results: Gamma-irradiation had a negative influence on the average plant height, leaf length, leaf width, and fresh weight. Photosynthetic pigment levels decreased with increasing dosages of gamma-irradiation, while soluble sugar content slightly increased. Gamma-irradiation responsive proteins were detected and identified by two-dimensional gel electrophoresis (2D-PAGE) and matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF). The proteins had a role in photosynthetic carbon fixation, anabolic pathway glycolysis, mitochondrial ATP production, and oxidative stress response regulation. MDA levels and activities of antioxidant enzymes such as superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT), and peroxidase (POD) increased with the increase in gamma-irradiation dosage level., Conclusions: This study provides some basic information regarding responses to gamma-irradiation, and provides valuable physiological and biological data on the effects of different gamma-irradiation dosages on Triticeae species.

Purpose: To investigate the biological effects of titanium ((48)Ti, one of the important heavy ions found in space) in the liver of exposed-mice., Materials and Methods: We gave adult male CBA/CaJ mice a whole-body exposure to a total dose of 0, 0.1, 0.25 or 0.5 Gy of (48)Ti ions. The liver was collected at 1 week, 1 month, and 6 months post-irradiation (five mice per treatment-group at each harvest-time). Three biological endpoints were used for evaluating the effects of (48)Ti ions: Oxidative-stress, inflammatory responses, and DNA-methylation (5-methylcytosine and 5-hydroxymethylcytosine)., Results: Our data clearly demonstrated dose-dependent increases in oxidative stress and inflammatory responses in the liver of exposed mice at all time-points (Analysis of Variance or ANOVA, p < 0.05). Significant dose-dependent increases in the levels of 5-methylcytosine were detected at 1 week and 1 month (ANOVA, p < 0.05). At 6 months post-irradiation, a significant increase in the level of 5-methylcytosine was found only in 0.5-Gy-(48)Ti-ion-exposed mice. In contrast, dose-dependent decreases in 5-hydroxymethylcytosine levels were found in the liver of exposed mice (ANOVA, p < 0.05) at all time-points., Conclusions: Chronic oxidative-stress, chronic inflammation, and persistent aberrant DNA-methylation occurred in the liver of (48)Ti-exposed mice. Hence, exposure to (48)Ti ions in space may pose health risks.

Purpose: To investigate the effect of gamma radiation-induced alterations in antioxidant defence of radioresistant Sf9 insect cells., Materials and Methods: Sf9 cells were irradiated at doses ranging from 0.5-200 Gy. Lipid peroxidation and protein carbonylation levels were observed at 4 h post-exposure along with reduced glutathione/oxidized glutathione (GSH/GSSG) profile as well as specific activities of redox active enzymes superoxide dismutase (SOD), catalase, ascorbate peroxidase (APOx), and glutathione reductase (GR). Human brain malignant glioma (BMG-1) cells were used for comparing radiation response of mammalian cells., Results: Sf9 cells displayed significantly less radiation-induced reactive oxygen/nitrogen species (ROS/RNS) generation, protein carbonylation and growth inhibition as compared to mammalian cells. Sf9 cells have higher basal APOx (∼4-fold), catalase (∼1.7-fold), SOD (∼1.3-fold) activity and GSH level (∼2.2-fold) compared to mammalian cells. A radiation dose-dependent increase in SOD, Catalase and APOx activity was found in Sf9 cells at least up to 100 Gy dose, while maximum activity in mammalian cells was achieved by 10 Gy., Conclusion: The present study suggests that Lepidopteran insect cells carry a stronger antioxidant system that protects against radiation-induced macromolecular damage, growth inhibition and cell death.

Purpose: To investigate the extent of γ-irradiation-induced oxidative membrane damage and antioxidant activity of quercetin in long-term, cold stored (4°C) acid-citrate-dextrose- preserved human red blood cells (RBC)., Materials and Methods: The extracellular activity of lactate dehydrogenase (LDH) was measured to assess RBC membrane integrity. Lipid peroxidation and reduced glutathione (GSH) levels were quantified by thiobarbituric acid-reactive substances (TBARS) and Ellman's reagent, respectively., Results: During storage of non-irradiated RBC (up 21 days) the LDH activity in the supernatant increased with time. In contrast to a low dose of ionizing radiation (30 Gy), irradiation at higher, but still clinically relevant doses, of 40-50 Gy resulted in elevation of the post-storage extracellular LDH activity. Quercetin (2-50 μM) dissolved in dimethyl sulfoxide (DMSO) significantly increased the LDH release in the irradiated and non-irradiated RBC, reflecting an increase of RBC membrane permeability. In the presence of ethanol as a solvent quercetin protected RBC against storage-induced oxidative damage - it inhibited the LDH release, GSH depletion, and lipid peroxidation., Conclusion: The level of protection offered by quercetin against the radiation- and storage-induced oxidative damage to RBC does not seem to be sufficient to warrant its application as an additive for conservation purposes. The findings indicate that the solvent can modulate a response of RBC to water-insoluble antioxidants changing their properties from anti-oxidative to pro-oxidative.

Purpose: To assess the effect of 950 MHz ultra-high-frequency electromagnetic radiation (UHF EMR) on biomarkers of oxidative damage, as well as to verify the concentration of unsaturated fatty acids (UFA) and the expression of the catalase in the livers of rats of different ages., Materials and Methods: Twelve rats were equally divided into two groups as controls (CR) and exposed (ER), for each age (0, 6, 15 and 30 days). Radiation exposure lasted half an hour per day for up to 51 days (21 days of gestation and 6, 15 or 30 days of life outside the womb). The specific absorption rate (SAR) ranged from 1.3-1.0 W/kg. The damage to lipids, proteins and DNA was verified by thiobarbituric acid reactive substances (TBARS), protein carbonyls and comets, respectively. UFA were determined by gas chromatography with a flame ionization detector. The expression of catalase was by Western blotting., Results: The neonates had low levels of TBARS and concentrations of UFA after exposure. There was no age difference in the accumulation of protein carbonyls for any age. The DNA damage of ER 15 or 30 days was different. The exposed neonates exhibited lower expression of catalase., Conclusions: 950 MHz UHF EMR does not cause oxidative stress (OS), and it is not genotoxic to the livers of neonates or those of 6 and 15 day old rats, but it changes the concentrations of polyunsaturated fatty acid (PUFA) in neonates. For rats of 30 days, no OS, but it is genotoxic to the livers of ER to total body irradiation.

Purpose: Radioprotection and mitigation effects of the antioxidants, Eukarion (EUK)-207, curcumin, and the curcumin analogs D12 and D68, on radiation-induced DNA damage or lipid peroxidation in murine skin were investigated. These antioxidants were studied because they have been previously reported to protect or mitigate against radiation-induced skin reactions., Methods: DNA damage was assessed using two different assays. A cytokinesis-blocked micronucleus (MN) assay was performed on primary skin fibroblasts harvested from the skin of C3H/HeJ male mice 1 day, 1 week and 4 weeks after 5 Gy or 10 Gy irradiation. Local skin or whole body irradiation (100 kVp X-rays or caesium (Cs)-137 γ-rays respectively) was performed. DNA damage was further quantified in keratinocytes by immunofluorescence staining of γ-histone 2AX (γ-H2AX) foci in formalin-fixed skin harvested 1 hour or 1 day post-whole body irradiation. Radiation-induced lipid peroxidation in the skin was investigated at the same time points as the MN assay by measuring malondialdehyde (MDA) with a Thiobarbituric acid reactive substances (TBARS) assay., Results: None of the studied antioxidants showed significant mitigation of skin DNA damage induced by local irradiation. However, when EUK-207 or curcumin were delivered before irradiation they provided some protection against DNA damage. In contrast, all the studied antioxidants demonstrated significant mitigating and protecting effects on radiation-induced lipid peroxidation at one or more of the three time points after local skin irradiation., Conclusion: Our results show no evidence for mitigation of DNA damage by the antioxidants studied in contrast to mitigation of lipid peroxidation. Since these agents have been reported to mitigate skin reactions following irradiation, the data suggest that changes in lipid peroxidation levels in skin may reflect developing skin reactions better than residual post-irradiation DNA damage in skin cells. Further direct comparison studies are required to confirm this inference from the data.

Purpose: The present study was undertaken to evaluate the radioprotective effect of meso-zeaxanthin, a xanthophyll carotenoid with profound antioxidant activity., Materials and Methods: Swiss albino mice were treated with different doses of meso-zeaxanthin (50 and 250 mg/kg body weight, orally) five days before irradiation and sacrificed at different time points. The protective effects of meso-zeaxanthin on mortality, haematological parameters, bone marrow cellularity and gastrointestinal system of irradiated mice were studied. The anti-genotoxic action of meso-zeaxanthin was studied by measuring micronuclei formation, chromosomal aberrations and DNA damage (comet assay)., Results: Meso-zeaxanthin administration significantly increased the lifespan of irradiated mice and reduced myelosuppression as evident from increases in white blood cell counts, bone marrow cellularity and the number of maturing monocytes when compared to the myelosuppression in radiation control animals. Meso-zeaxanthin significantly elevated the radiation-induced reduction in the activities of superoxide dismutase, catalase, glutathione peroxidase and glutathione in both liver and intestinal mucosa. The carotenoid-treated animals showed a profound reduction in genotoxic activity which was apparent in decreases in micronuclei formation and chromosomal aberrations. Irradiation also induced damage to cellular DNA as was obvious from increases in comet parameters like tail DNA%, tail moment, tail length and Olive tail moment in the radiation control group. These parameters were decreased by meso-zeaxanthin treatment., Conclusion: Results indicated a radioprotective potential of meso-zeaxanthin.

Unlabelled: Abstract Purpose: The aim of the present study was to evaluate the electrophysiological, biochemical and ultrastructural changes on the rat sciatic nerve after radiotherapy., Material and Methods: Thirty male Wistar albino rats were divided into three groups as: Control group (n = 10), Group I: 3 months after radiotherapy (n = 10), and Group II: 6 months after radiotherapy (n = 10). Groups I and II were irradiated with a (60)Co gamma source. A dose of 20 Gy in 10 fractions was applied to Groups I and II. Compound motor action potentials (CMAP) were recorded in all groups. Superoxide dismutase (SOD) and catalase (CAT) activities and malondialdehyde (MDA) levels were measured in the sciatic nerve of rats using the biochemical methods. Ultrastructural changes were determined by electron microscopy., Results: In Groups I and II, the amplitude of CMAP was significantly lower and the latency was significantly higher than that of the control group. There were no significant differences between Groups I and II regarding the CMAP amplitude and latency. The MDA levels were significantly increased, whereas the SOD and CAT activities were significantly decreased in experimental groups when compared with the control group. However, there were no significant changes in these parameters between Groups I and II. Degeneration in myelinated nerve fibers was observed ultrastructurally only in the experimental groups. Significant changes were observed between the control group and experimental groups in terms of ultrastructural myelin grading score and axonal damage score. No significant differences were found between Groups I and II., Conclusions: These findings indicated that the dose of 20 Gy in 10 fractions radiotherapy caused neuropathic damages in normal rat sciatic nerve 3 and 6 months after irradiation.

Purpose: This study was conducted to evaluate the effect of radiofrequency wave (RFW)-induced oxidative stress in the eye and the prophylactic effect of vitamin C on this organ by measuring the antioxidant enzymes activity including: glutathione peroxidase (GPx), superoxide dismutase (SOD) and catalase (CAT), and malondialdehyde (MDA)., Materials and Methods: Thirty-two adult male Sprague-Dawley rats were randomly divided into four experimental groups and treated daily for 45 days as follows: Control, vitamin C (L-ascorbic acid 200 mg/kg of body weight/day by gavage), test (exposed to 900 MHz RFW) and the treated group (received vitamin C in addition to exposure to RFW). At the end of the experiment all animals were sacrificed, their eyes were removed and were used for measurement of antioxidant enzymes and MDA activity., Results: The results indicate that exposure to RFW in the test group decreased antioxidant enzymes activity and increased MDA compared with the control groups (P < 0.05). In the treated group vitamin C improved antioxidant enzymes activity and reduced MDA compared to the test group (P < 0.05)., Conclusions: It can be concluded that RFW causes oxidative stress in the eyes and vitamin C improves the antioxidant enzymes activity and decreases MDA.

Purpose: To investigate the extent of γ-irradiation-induced oxidative protein and lipid damage in long-term (up to 21 days) cold stored (4°C) erythrocytes (RBC) and in plasma from whole blood anticoagulated with acid-citrate-dextrose (ACD-A)., Materials and Methods: Lipid peroxidation, protein carbonyl group (CO) and thiol levels were quantified by the amount of thiobarbituric acid-reactive substances (TBARS), enzyme-linked immunosorbent assay (ELISA) and with Ellman reagent, respectively., Results: Irradiation (40-50 Gy) enhanced lipid peroxidation in the RBC membrane (at day 1 and after 21 storage days); the increase was storage time-dependent. In pre-irradiated (30-50 Gy) and long-term stored RBC membrane protein CO level was higher vs. non-irradiated. Irradiation resulted in RBC membrane protein thiol level elevation, most likely being a result of conformational changes and/or the polypeptide chain fragmentation. Similar to RBC, irradiation of plasma resulted in the increased TBARS generation. In plasma, significant protein CO elevation (at dose of 50 Gy) and protein thiol reduction (30-50 Gy) was observed., Conclusion: These findings clearly indicate that irradiation at clinically relevant doses enhances the degree of lipid peroxidation and oxidative protein damage in the membranes of stored RBC. The oxidative stress markers may be considered as additional parameters for RBC quality assessment in the blood banks.

Purpose: Electromagnetic radiation from wireless devices may affect biological systems by increasing free radicals. The present study was designed to determine the effects of 2.45 GHz radiation on the antioxidant redox system, calcium ion signaling, cell count and viability in human leukemia 60 cells., Materials and Methods: Twelve cell cultures were equally divided into two main groups as controls (n = 6) and irradiated (n = 6) and then subdivided into four different subgroups depending on the duration of exposure, namely 1, 2, 12 and 24 hours. The samples were analyzed immediately after the experimental period., Results: The extent of lipid peroxidation, cytosolic free Ca²⁺ and cell numbers were higher in 2.45 GHz groups than in the controls. The increase of cytosolic free Ca²⁺ concentrations was radiation time-dependent and was highest at 24-h exposure. The reduced glutathione, glutathione peroxidase, vitamin C and cell viability values did not show any changes in any of the experimental groups. 2-aminoethyl diphenylborinate inhibits Ca²⁺ ions influx by blockage of the transient receptor potential melastatin 2., Conclusions: 2.45 GHz electromagnetic radiation appears to induce proliferative effects through oxidative stress and Ca²⁺ influx although blocking of transient receptor potential melastatin 2 channels by 2-aminoethyl diphenylborinate seems to counteract the effects on Ca²⁺ ions influx.

Purpose: Environmental electromagnetic fields originate from man-made sources, such as mobile phones and base stations, and have led to increasing public concern about their possible adverse health effects. We aimed to investigate the possible effects of radiofrequency radiation (RFR) generated from these devices on oversensitive animals, such as pregnant rabbits., Materials and Methods: In the present study, the effects of whole body 1800 MHz Global System for Mobile Communications (GSM)-like RFR exposure for 15 min/day for seven days on blood chemistry and lipid peroxidation levels in both non-pregnant and pregnant New Zealand White rabbits were investigated. Thirteen-month-old rabbits were studied in the following four groups: Non-pregnant control, non-pregnant RFR-exposed, pregnant control and pregnant RFR-exposed., Results: Lipid peroxidation, namely malondialdehyde (MDA) levels, did not change after RFR exposure. However, blood chemistry parameters, such as cholesterol (CHO), total protein (TP), albumin (ALB), uric acid, creatinin and creatine kinase (CK) and creatine kinase-myocardial band isoenzyme (CK-MB) changed due to both pregnancy and RFR exposure., Conclusion: Our investigations have been shown that no indication for oxidative stress was detected in the blood of pregnant rabbits upon RF exposure at specific conditions employed in the present study. Minor changes in some blood chemistry parameters were detected but CK-MB and CK increases were found remarkable. Studies on RFR exposure during pregnancy will help establish international standards for the protection of pregnant women from environmental RFR.

Purpose: We aimed to design a prolonged radiofrequency (RF) radiation exposure and investigate in an animal model, possible bio-effects of RF radiation on the ongoing developmental stages of children from conception to childhood., Materials and Methods: A total of 72 New Zealand female and male white rabbits aged one month were used. Females were exposed to RF radiation for 15 min/day during 7 days, whereas males were exposed to the same level of radiation for 15 min/day during 14 days. Thirty-six female and 36 male infant rabbits were randomly divided into four groups: Group I [Intrauterine (IU) exposure (-); Extrauterine (EU) exposure (-)]: Sham exposure which means rabbits were exposed to 1800 MHz Global System for Mobile Telecommunication (GSM)-like RF signals neither in the IU nor in the EU periods. Group II [IU exposure (-); EU exposure (+)]: Infant rabbits were exposed to 1800 MHz GSM-like RF signals when they reached one month of age. Group III [IU exposure (+); EU exposure (-)]: Infant rabbits were exposed to 1800 MHz GSM-like RF signals in the IU period (between 15th and 22nd days of the gestational period). Group IV [IU exposure (+); EU exposure (+)]: Infant rabbits were exposed to 1800 MHz GSM-like RF signals both in the IU period (between 15th and 22nd days of the gestational period) and in the EU period when they reached one month of age. Biochemical analysis for lipid peroxidation and DNA damage were carried out in the livers of all rabbits., Results: Lipid peroxidation levels in the liver tissues of female and male infant rabbits increased under RF radiation exposure. Liver 8-hydroxy-2'-deoxyguanosine (8-OHdG) levels of female rabbits exposed to RF radiation were also found to increase when compared with the levels of non-exposed infants. However, there were no changes in liver 8-OHdG levels of male rabbits under RF exposure., Conclusion: Consequently, it can be concluded that GSM-like RF radiation may induce biochemical changes by increasing free radical attacks to structural biomolecules in the rabbit as an experimental animal model.

Purpose: To investigate the effects of electromagnetic pulses (EMP) on associative learning in mice and test a preliminary mechanism for these effects., Materials and Methods: A tapered parallel plate gigahertz transverse electromagnetic (GTEM) cell with a flared rectangular coaxial transmission line was used to expose male BALB/c mice to EMP (peak-intensity 400 kV/m, rise-time 10 ns, pulse-width 350 ns, 0.5 Hz and total 200 pulses). Concurrent sham-exposed mice were used as a control. Associative learning, oxidative stress in the brain, serum chemistry and the protective action of tocopherol monoglucoside (TMG) in mice were measured, respectively., Results: (1) Twelve hour and 1 day post EMP exposure associative learning was reduced significantly compared with sham control (p<0.05) but recovered at 2 d post EMP exposure. (2) Compared with the sham control, lipid peroxidation of brain tissue and chemiluminescence (CL) intensity increased significantly (p<0.05), while the activity of the antioxidant enzymes Superoxide Dismutase [SOD], Glutathione [GSH], Glutathione Peroxidase [GSH-Px], Catalase [CAT]) decreased significantly (p<0.05) at 3 h, 6 h, 12 h and 1 d post EMP exposure. All these parameters recovered at 2 d post EMP exposure. (3) No significant differences between the sham control group and EMP exposed group were observed in serum cholesterol and triglycerides. (4) Pretreatment of mice with TMG showed protective effects to EMP exposure., Conclusions: EMP exposure significantly decreased associative learning in mice and TMG acted as an effective protective agent from EMP exposure. This mechanism could involve an increase of oxidative stress in brain by EMP exposure.

Purpose: The aim of the present study was to evaluate the early effects of acute (2 h) exposure to extremely low frequency electromagnetic fields (ELF-EMF), as well as movement restraint (MR) and the combination of both on the antioxidant systems in the plasma, liver, kidney, and heart of rats., Materials and Methods: Twenty-four adult male Wistar rats were divided in two groups, restrained and unrestrained. The restrained animals were confined into an acrylic tube for 120 min. Half of the animals of each group were exposed to ELF-EMF (60 Hz, 2.4 mT) during the period of restriction. Immediately after treatment, reduced glutathione (GSH), catalase (CAT), superoxide dismutase (SOD), and thiobarbituric acid reactive substances (TBARS) were measured in tissues., Results: GSH concentration was significantly lower in the heart of all experimental animals when compared to the control group; furthermore, the decrease was higher in the liver of restrained animals. SOD activity was lower in the plasma of restrained and EMF exposed animals compared to unrestrained rats. There were no significant differences in CAT activity and TBARS levels among all the experimental groups vs. the control group., Conclusion: Two hours of 60 Hz EMF exposure might immediately alter the metabolism of free radicals, decreasing SOD activity in plasma and GSH content in heart and kidney, but does not induce immediate lipid peroxidation. Oxidative stress induced by movement restraint was stronger than that produced by EMF.

Purpose: Ultraviolet C (UVC) irradiation of aqueous solutions is known to be a good source of reactive oxygen species (ROS). The aim of this study is to examine the effect of increasing doses of UVC irradiation, in the presence and absence of the antioxidant butylated hydroxytoluene (BHT), on human sperm motility and lipid peroxidation of its membranes., Materials and Methods: Human sperm samples were irradiated with UVC light (254 nm) for different periods of time. A computer-assisted semen analysis of sperm motility was carried out after UV irradiation. The percentage of motile sperm (%MOT), progressive motility, straight line velocity (VSL), curvilinear velocity (VCL) and the percentage of linearity (%LIN) were evaluated. The level of lipid peroxidation of sperm membranes was estimated by measurement of the thiobarbituric acid reactive substances (TBARS)., Results: UVC irradiation of human spermatozoa produced a diminution of the sperm motility (%MOT, progressive motility, VSL, VCL, %LIN), viability and, concomitantly, an increase of the level of lipid peroxidation of the sperm membranes. The observed effects of the UVC irradiation were prevented by addition of the antioxidant BHT, indicating that the effects of UVC on the tested sperm parameters are mediated by an important rise in lipid peroxidation of the sperm membrane., Conclusion: Lipid peroxidation of the human sperm plasma membrane leads to a decrease in the sperm motility (%MOT, progressive motility, VSL, VCL, %LIN) and viability. The protective effect of BHT on the UVC-irradiated sperm cells indicates the effects of ROS on sperm function.

Purpose: To study the biochemical, quantitative histopathological and behavioural changes after 5 Gy whole body irradiation and its modulation by supplementation of Grewia asiatica extract (GAE) on male Swiss albino mice up to 30 days., Materials and Methods: For this, healthy mice from an inbred colony were divided into five groups: (i) Control (vehicle treated); (ii) GAE treated--mice in this group were orally supplemented with GAE (700 mg/kg body weight (bw)/day) once daily for 15 consecutive days; (iii) Irradiated mice; (iv) GAE + irradiated--mice in this group were orally supplemented GAE for 15 days (once a day) prior to irradiation; and (v) Irradiated + GAE--mice in this group were administered GAE orally for 15 days (once a day) consequently after irradiation., Results: Marked radiation induced changes in the amount of cerebellar lipid peroxidation (LPO), Glutathione (GSH), protein, nucleic acids and histopathological changes could be significantly (p < 0.001) ameliorated specially at later intervals by supplementation of GAE prior to and post irradiation. Radiation induced deficits in learning and memory were also significantly (p < 0.001) ameliorated., Conclusion: The result of present study showed that prior/post-supplementation of G. asiatica has radioprotective potential as well as neuroprotective properties against the radiation.

Purpose: Electromagnetic radiation (EMR) from wireless devices may affect biological systems by increasing free radicals. The present study was designed to determine the effects of 2.45 GHz EMR on the brain antioxidant redox system and electroencephalography (EEG) records in rat. The possible protective effects of selenium and L-carnitine were also tested and compared to untreated controls., Materials and Methods: Thirty rats were equally divided into five different groups, namely Group A(1): Cage control, Group A(2): Sham control, group B: 2.45 GHz EMR, group C: 2.45 GHz EMR + selenium, group D: 2.45 GHz EMR + L-carnitine. Groups B, C and D were exposed to 2.45 GHz EMR during 60 min/day for 28 days. End of the experiments, EEG records and the brain cortex samples were taken., Results: The cortex brain vitamin A (p < 0.05), vitamin C (p < 0.01) and vitamin E (p < 0.05) concentrations values were lower in group B than in group A1 and A2 although their concentrations were increased by selenium and L-carnitine supplementation. Lipid peroxidation, levels were lower in group C (p < 0.05) and D (p < 0.01) than in group B where as reduced glutathione levels were higher in group C (p < 0.05) than in group A1, A2 and B. However, B-carotene levels did not change in the five groups., Conclusions: L-carnitine and selenium seem to have protective effects on the 2.45 GHz-induced decrease of the vitamins by supporting antioxidant redox system. L-carnitine on the vitamin concentrations seems to more protective affect than in selenium.

Purpose: Thorium ((232)Th), a heavy metal radionuclide that targets the liver and skeleton, has been shown to accumulate in the central nervous system at low levels. The present study was aimed to investigate neurobehavioural and neurochemical changes in mice treated with (232)Th at sub-lethal doses., Materials and Methods: Swiss albino mice were administered intraperitoneally with thorium nitrate. The chelation-based therapeutic effect of calcium diethylenetriamine pentaacetate (Ca-DTPA) was tested on the (232)Th-treated mice. (232)Th localisation was determined in brain regions by the Inductively Coupled Plasma-Atomic Emission Spectroscopy (ICP-AES) method. Achetylcholine esterase (AChE) activity in different brain regions was evaluated to assess the cholinergic function of mice CNS. Oxidative damage was evaluated by assessing the activities of antioxidant enzymes (i.e., superoxide dismutase and catalase) and the level of lipid peroxidation. The neurobehavioural alteration in the treated mice was studied by the shuttle box method., Results: (232)Th accumulation found in different brain regions followed the order: Cerebellum (Cbl) > cortex (Ctx) > hippocampus (Hp) > striatum (Str). However, removal of (232)Th by Ca-DTPA was significant from brain regions like Cbl, Ctx and Str but not from Hp. A significant increase in lipid peroxidation and acetylcholine esterase (AChE) activity was observed in the treated mice but activities of superoxide dismutase and catalase was found substantially decreased. (232)Th treatment impaired the learning and memory-based neurobehaviour of the mice. Furthermore, our data suggest that Ca-DTPA injection in (232)Th-treated animals failed to improve the neurobehaviour of the treated mice, perhaps because Ca-DTPA could not decorporate (232)Th or mitigate (232)Th-mediated neurochemical changes effectively from/in hippocampus, a brain region implicated in learning and memory response., Conclusion: Administration of (232)Th in mice caused neurobehavioural alteration and impairment of cholinergic function, which might be the consequence(s) of oxidative stress induction in different brain regions.

Purpose: UVA radiation (315-400 nm) contributes to skin aging and carcinogenesis. The aim of this review is to consider the mechanisms that underlie UVA-induced cellular damage, how this damage may be prevented or repaired and the signal transduction processes that are elicited in response to it., Results: Exposure to ultraviolet (UV) light is well-established as the causative factor in skin cancer. Until recently, most work on the mechanisms that underlie skin carcinogenesis focused on shorter wavelength UVB radiation (280-315 nm), however in recent years there has been increased interest in the contribution made by UVA. UVA is able to cause a range of damage to cellular biomolecules including lipid peroxidation, oxidized protein and DNA damage, such as 8-oxoguanine and cyclobutane pyrimidine dimers. Such damage is strongly implicated in both cell death and malignant transformation and cells have a number of mechanisms in place to mitigate the effects of UVA exposure, including antioxidants, DNA repair, and stress signalling pathways., Conclusions: The past decade has seen a surge of interest in the biological effects of UVA exposure as its significance to the process of photo-carcinogenesis has become increasingly evident. However, unpicking the unique complexity of the cellular response to UVA, which is only now becoming apparent, will be a major challenge for the field of photobiology in the 21st century.

Purpose: Biological effects of X-ray irradiation and protection by melatonin on the immune status of a tropical rodent, Funambulus pennanti, was examined by estimating oxidative damage of peripheral blood and spleen and protection by melatonin treatment., Material and Methods: Seventy squirrels were divided into 4 sets of 25, 25, 10 and 10 having subdivisions into a total of eight groups. Squirrels of set 1 (groups A and B) received only normal saline, set 2 (groups C and D) received 25 microg/100 g body weight (bwt) melatonin and set 3 (groups E and F) received 25 microg/100 g bwt vitamin E for four weeks. Groups G and H of set 4 received high doses of melatonin (0.5 mg/100 g bwt) 30 min prior to and 30 min after X-ray irradiation, respectively, and were sacrificed 1 h after irradiation. Groups B and D were sacrificed after 4 h, 24 h, 48 h and 72 h of irradiation for total leukocyte count (TLC) in peripheral blood, percent apoptotic cells and lipid peroxidation (LPO) in spleen while group F was sacrificed after 4 h of irradiation to measure LPO., Results: Pre-melatonin treatment (25 microg/100 g bwt) restored TLC, percent apoptotic cells and LPO levels of X-ray exposed squirrels. High dose pre-melatonin treatment (0.5 mg/100 g bwt) restored the above conditions significantly while post treatment did not. Vitamin E reduced elevated LPO level in irradiated tissue, but the effect of melatonin was more potent., Conclusion: Melatonin administration prior to X-ray irradiation prevented radiation-induced oxidative damage during the reproductively active phase of the seasonally breeding rodent suggesting a high protective role of melatonin following X-ray irradiation.

Purpose: To evaluate the radiomodulatory influence of a leaf extract of Mentha piperita (Linn) on hepatic antioxidant status and lipid peroxidation in Swiss albino mice., Materials and Methods: Animals were given either double distilled water or leaf extract of M. piperita orally (1 g/kg bwt/day) once a day for three consecutive days. Thirty min after the last treatment mice were exposed to 8 Gy of gamma radiation. Mice were autopsied at 30 min post-irradiation. Biochemical parameters were studied to assess the radioprotective effect of leaf extract of M. piperita., Results: Animals pretreated with leaf extract of M. piperita and exposed to 8.0 Gy gamma radiation showed a significant increase in the activities of reduced glutathione content (p < 0.001), glutathione peroxidase (p < 0.005), glutathione reductase (p < 0.001), glutathione S-transferase (p < 0.005), superoxide dismutase (p < 0.005), and catalase (p < 0.005). Irradiated group pretreated with leaf extract of M. piperita showed significant decrease in malondialdehyde (MDA) formation in liver. The leaf extract of M. piperita showed strong radical scavenging activity in both the 1, 1 diphenyl-2-picryl hydrazyl radical (DPPH*) and 2, 2 azinobis (3-ethylbenzothiazoline-6-sulfonic acid) radical cation (ABTS*+) assays., Conclusions: The results of the present investigation suggest the antioxidant and free radical scavenging activities of leaf extract of M. piperita are the likely mechanism of radiation protection.

Purpose: To investigate changes in radical scavenging ability and lipid peroxidation in liver microsomal membranes and cooperative suppression of lipid peroxidation by microsomal and cytosolic radical scavengers, 24 h after whole-body, low-dose X-irradiation of rats., Materials and Methods: Male Wistar rats were irradiated with 1-50 cGy of X-rays. Liver microsomal radical scavenging ability was determined using the trapping ability of 1,1-diphenyl-2-picrylhydrazyl (DPPH), a stable free radical. Microsomal alpha-tocopherol (Vit.E) content was determined using an electrochemical detector. Microsomal glutathione peroxidase (GPx) activity was determined as the consuming rate of NADPH. Microsomal lipid peroxidation was determined by the thiobarbituric acid method., Results: Low molecular weight radical scavenging ability of rat liver microsomes increased 24 h after whole-body, low-dose X-irradiation when alpha-tocopherol was included, showing a maximum level at 5-10 cGy. Microsomal GPx activity also increased 24 h after 5 cGy irradiation. The lipid peroxidation level in microsomes decreased, showing a maximal suppression at 5 cGy. High-dose irradiation-induced microsomal lipid peroxidation was strongly suppressed cooperatively by microsomal and cytosolic antioxidants induced by low-dose irradiation., Conclusion: Low doses of radiation induce increases in liver microsomal antioxidants, which in turn result in enhanced suppression of microsomal lipid peroxidation cooperatively with cytosolic antioxidants induced by low-dose irradiation.

Purpose: To investigate the regulation of NADPH-producing isocitrate dehydrogenase (ICDH) in cytosol (IDPc) and mitochondria (IDPm) upon gamma-ray irradiation, and the roles of IDPc and IDPm in the protection against cellular damage induced by gamma-ray irradiation., Materials and Methods: Changes of IDPc and IDPm proteins upon gamma-ray irradiation to NIH3T3 cells were analysed by immunoblotting. To increase or decrease the expression of IDPc or IDPm, NIH3T3 cells were stably transfected with mouse IDPc or IDPm cDNA in either the sense or the antisense direction. The transfected cells with either increased or decreased IDPc or IDPm were exposed to gamma-rays, and the levels of reactive oxygen species generation, protein oxidation and lipid peroxidation were measured., Results: Both IDPc and IDPm activities were induced by gamma-ray in NIH3T3 cells. Cells with decreased expression of IDPc or IDPm had elevated reactive oxygen species generation, lipid peroxidation and protein oxidation. Conversely, overproduction of IDPc or IDPm protein partially protected the cells from oxidative damage induced by gamma-ray irradiation., Conclusions: The protective role of IDPc and IDPm against gamma-ray-induced cellular damage can be attributed to elevated NADPH, reducing equivalents needed for recycling reduced glutathione in the cytosol and mitochondria. Thus, a primary biological function of the ICDHs may be production of NADPH, which is a prerequisite for some cellular defence systems against oxidative damage.

Purpose: To address the links between calcium, peroxidation, cell damage and death and the response of the enzymes involved in free radical metabolism, in splenocytes of mice irradiated with gamma-rays., Materials and Methods: Splenocytes of Swiss albino mice were irradiated with various doses (0-7 Gy) of gamma-rays (60Co) at a dose-rate of 0.0575 Gy s(-1). Membrane peroxidation and fluidity were determined by the thiobarbituric acid-reactive substances (TBARS) method, and fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene (DPH), respectively. Apoptosis was analysed by nucleosomal ladder formation and activity of NF-kappaB by electrophoretic mobility shift assay (EMSA). The specific activities of the antioxidant enzymes, lactate dehydrogenase (LDH), levels of nitric oxide (NO*) and glutathione were determined spectrophotometrically. Modulatory effects of Ca2+ were examined at 3 Gy using different concentrations (1, 3 and 5 mM) in the presence or absence of the ionophore A23187., Results: Irradiation of splenocytes resulted in enhanced peroxidative damage. membrane fluidity, apoptosis and DNA binding activity of NF-kappaB. The specific activities of LDH and antioxidant enzymes superoxide dismutase (SOD), DT-diaphorase (DTD), glutathione S-transferase (GST) and levels of glutathione (GSH) and NO* were increased with radiation dose up to 4Gy. Ca2+ augmented the radiation-induced responses. The presence of ionophore A23187 potentiated the modulatory effects of Ca2+., Conclusions: These findings show that Ca2+ augments radiation damage and is more effective intracellularly. Ca2+, peroxidation, cellular damage and apoptosis are possibly interlinked through signals, as is evident from the increased activity of NF-kappaB and generation of NO*. The enhanced antioxidant status suggests an attempt made by the irradiated cells to maintain their normal functions.

Purpose: To ascertain the possibility of fragmentation processes in lipid membranes when acted upon with gamma-radiation under various conditions, and to evaluate fragmentation processes quantitatively in comparison with the lipid peroxidation processes., Materials and Methods: Phospholipids as components of multilamellar liposomes exposed to 137Cs gamma-rays at dose-rates of 0.06 and 0.33 Gy s-1 in the dose range 0-18 kGy. Peroxidation products were determined spectrophotometrically and fragmentation products were analysed using thin-layer chromatography and GLC methods., Results: Phosphatidylglycerol containing unsaturated fatty acid residues and free hydroxyl groups underwent both peroxidation and fragmentation when its aqueous dispersions were treated with ionizing radiation. gamma-Radiolysis of multilamellar liposomes led to the formation of peroxidation products, as well as phosphatidic acid and hydroxyacetone, fragmentation products of the initial lipid. Comparable radiation-chemical yields of conjugated-diene products and phosphatidic acid indicate that the probabilities of oxidation and fragmentation processes to occur are approximately equal in this case. In contrast to lipid peroxidation, lipid fragmentation is not accelerated but suppressed by oxygen and appears to be a prevailing process in de-aerated solutions. The phosphatidic acid formed as a result of phosphatidylglycerol fragmentation can intensify the lipid peroxidation., Conclusions: Treatment of lipid membranes with ionizing radiation leads to the destructive processes within both hydrophobic and hydrophilic moiety of lipids.

Purpose: Following the demonstration that caffeine effectively competes with oxygen for electrons and also scavenges hydroxyl radicals and singlet oxygen, the differential modification of oxygen-dependent and independent effects of gamma-radiation by caffeine in membranes was examined, using rat liver mitochondria as a model system., Materials and Methods: Mitochondria were isolated from the livers of Wistar rats and exposed to gamma-radiation in the dose range of 45-600 Gy (dose rate 15 Gy/min) in the presence or absence of caffeine. To examine the 'oxygen effect', post-irradiation incubation was carried out in the presence of oxygen or nitrogen in buffers saturated with the respective gases. Membrane damage was examined as lipid peroxidation (assessed as formation of thiobarbituric acid-reactive substances (TBARS), lipid hydroperoxides (LOOH) and conjugated dienes (CD), protein oxidation, depletion of protein thiols, superoxide dismutase or glutathione., Results: Lipid peroxidation increased as a function of radiation dose, from 45 to 600 Gv. Post-irradiation incubation of mitochondria under nitrogen decreased the response, while incubation under oxygen saturation enhanced it significantly. The presence of caffeine during radiation exposure inhibited lipid peroxidation significantly as a function of concentration, in the range of 5 microM to 4 mM. The inhibition was highest with 4 mM of caffeine. Under oxic conditions, inhibition at 1 mM was significantly more than under anoxia. Anoxia was either ineffective or marginally increased peroxidation in the presence of caffeine. A similar observation was obtained when membrane damage was assessed as protein oxidation. Radiation-induced depletion of protein thiols was greatly enhanced by oxygen saturation and this was completely prevented by caffeine. This compound also protected against the radiation-induced loss of the antioxidant glutathione and the enzyme superoxide dismutase., Conclusions: The results suggest that caffeine effectively protected membranes against the oxic component of damage but may not do so for the anoxic component.

Purpose: Radical-chain mechanisms such as lipid peroxidation are known to show an inverse dose-rate effect, i.e. the radiation effect increases with decreasing dose rate at identical doses applied. The present study was intended to investigate whether an inverse dose-rate effect can be transferred from the level of lipids to the level of proteins., Method: Functional inactivation or structural modification by 80kV X-rays of two classes of proteins was investigated: membrane proteins with a natural environment of lipids like the Ca-ATPase of the sarcoplasmic reticulum, succinate dehydrogenase and F0F1-ATPase from the inner mitochondrial membrane. The second class comprises the water-soluble proteins cytosolic creatine kinase (MM-CK) and bovine serum albumin (BSA). Their modification by free radicals of water radiolysis was investigated in the absence and presence of lipid vesicles., Results: For all proteins investigated, an inverse dose-rate effect was observed in the presence of lipids. This also holds for the water-soluble proteins MM-CK and BSA. In the latter two cases, the dose-rate effect disappeared either in the absence of (unsaturated) lipids or in the presence of alpha-tocopherol., Conclusion: The largely identical results obtained for a variety of different proteins indicate that inverse dose-rate effects are a normal consequence of radiation induced protein damage in the presence of lipids. In view of the high amount of cellular lipids, this should also hold for the situation in vivo, although due to the comparatively high concentration of intracellular antioxidants the dose-rate dependence might be strongly reduced or even virtually abolished.

Purpose: To assess the roles of cell membranes and DNA as targets in radiation-induced apoptosis., Materials and Methods: Peripheral blood lymphocytes from normal human donors were exposed to different types of apoptosis-inducing agents. Several measures of apoptosis were used to compare the kinetics of the processes induced, as well as to correlate the processes with DNA damage and membrane oxidation., Results: Two kinetically distinct processes were observed. DNA-damaging agents, such as ionizing radiation, bleomycin, cisplatin and the topoisomerase inhibitor m-amsacrine, induced apoptosis by a kinetically slow process initiated by DNA damage and dependent on protein synthesis, but which did not correlate with membrane oxidation. Conversely, the agents t-butyl hydroperoxide and cumene hydroperoxide induced apoptosis by a kinetically fast process independent of protein synthesis and which did correlate with membrane oxidation., Conclusions: Slowly repaired or unrepairable DNA lesions, such as some of those produced by ionizing radiation exposure, trigger apoptosis by a kinetically slow process. This slow apoptotic pathway is distinct from a fast process not induced by radiation but which is induced by membrane-oxidizing agents.

Purpose: The investigation of the effects of ionizing radiation on hepatic cholesterol metabolism and the concentration and composition of plasma lipoproteins in the male Syrian hamster., Materials and Methods: After sublethal whole-body 60Co gamma-irradiation (8 Gy, 1 Gy/min), plasma lipoproteins were separated by density-gradient ultracentrifugation. Activities of hydroxymethylglutarylCoA (HMGCoA) reductase and of cholesterol 7alpha-hydroxylase were measured in hepatic microsomes and the low-density lipoprotein (LDL) receptor mass was determined in hepatic total membranes. Lipid peroxidation in LDL was assessed in vitro as the formation of conjugated dienes at 234 nm. A group of pair-fed animals served as controls as the food intake was markedly decreased with exposure to radiation., Results: Plasma lipid concentrations decreased 2 days post-irradiation and then markedly increased by day 6 post-irradiation; plasma cholesterol was increased by 77% and triglycerides by +207%. LDL accumulated in plasma while high-density lipoprotein (HDL) levels decreased. HDL contained significant amounts of apo SAA, the acute phase apolipoprotein. The activities of hepatic HMGCoA reductase, the rate-limiting enzyme for cholesterol synthesis, increased (+125%, p=0.06); hepatic cholesterol 7alpha-hydroxylase, the rate-limiting enzyme for bile acid synthesis, decreased (-85%); and the hepatic LDL receptor mass also decreased (-44%). The susceptibility of LDL to oxidation was also increased when animals were exposed to radiation., Conclusions: Lipoprotein modifications that appeared following radiation exposure may result from an induced inflammatory state and may further contribute to vascular damage.

Purpose: Phenothiazine drugs have been found to sensitize hypoxic cancer cells while offering protection to normal cells. Since phenothiazines are known to induce the cytochrome P450 system, its radiomodulation by phenothiazines has been examined., Materials and Methods: Mice were administered phenothiazines intraperitoneally and irradiated with different doses of gamma-rays at 1.38 Gy/min. The activities of NADPH-cytochrome P450 reductase and NADH-cytochrome b5 reductase, the content of cytochrome P450 and b5, the extent of lipid peroxidation as well as the activities of LDH, XO, SOD, GST and DTD were determined in the liver., Results: The levels of different components of the cytochrome P450 system and antioxidant enzymes were enhanced up to 5 Gy and decreased thereafter. However, a progressive increase was noticed in peroxidative damage and the activities of LDH and XO. Administration of phenothiazines enhanced the radiation effect on components of the cytochrome P450 system (except NADH-cytochrome b5 reductase) and the activities of SOD, GST and DTD. Concomitantly, phenothiazines inhibited lipid peroxidation, LDH and XO., Conclusions: Activation of the cytochrome P450 system by phenothiazines leading to the enhancement of antioxidant potential of animals and free-radical scavenging are attributes of the radioprotective action of phenothiazines.

Purpose: To investigate the effects of pre-exposure of mouse testis with low-doses of (16)O8+ ions or 60Co gamma-rays on sperm shape abnormalities, lipid peroxidation and superoxide dismutase (SOD) activity induced by subsequent high-dose irradiation., Materials and Methods: Testes of the B6C3F1 hybrid strain mice were pre-irradiated with 0.05 Gy of (16)O8+ ions or 60Co gamma-rays and then after 4 h given a test irradiation with 2 Gy of the same radiation type. SOD activity and thiobarbituric acid reactive substances (TBARS) in the testes were determined by spectrophotometric and TBA methods respectively at 4 h after irradiation. Testis weight, sperm count and sperm morphology were analysed at day 35 after irradiation., Results: Compared with controls, there was a significant increase in SOD activity and a significant decrease in TBARS level of pretreated testes. Testis weight loss, sperm count reduction and sperm abnormalities were significantly lower in the pretreated testes. The bioeffects of a 2 Gy dose of (16)O8+ ions relative to 60Co gamma-rays were 1.84 +/- 0.28 for testis weight, 1.22 +/- 0.25 for sperm count and 1.29 +/- 0.10 for sperm abnormalities., Conclusions: These data suggest that pre-exposure of testes with a low dose of heavy ions or gamma-rays renders the organ more resistant to subsequent high-dose irradiation. The increase of SOD activity and the decrease of lipid peroxidation levels induced by low-dose ionizing irradiation may be involved in this resistance. The effects with heavy ion irradiation were greater than with gamma-rays.

Fourier transform-infrared spectroscopy of dry, multibilayer films has been used to study gamma-radiation and UV-C light induced lipid peroxidation in 1,2-dilinoleoyl-sn-glycero-3-phosphocholine liposomes. The observed spectral changes were compared with the results obtained from measurement of hydroperoxides, conjugated dienes and to the formation of thiobarbituric acid reactive substances, such as malondialdehyde (MDA) or MDA-like substances. Upon irradiation a decrease in intensity of the asymmetric C - H stretching vibration (va(CH2)) of the isolated cis C = C - H groups (3010 cm-1) was observed. Directly correlated with the decrease of the va(CH2) absorption was a shift of the asymmetric phosphate ester stretching vibration (va(P = O)) towards smaller wavenumbers (1260-->1244 cm-1), indicating that the lipid peroxidation induced molecular alterations in the fatty acid chains influence the packing of the phospholipids in dry multibilayer films. In addition, the formation of a new absorption band at 1693 cm-1 could be detected, the intensity of which was comparable with the formation of thiobarbituric acid reactive substances and, therefore, attributed to the (C = O) stretching of alpha, beta unsaturated aldehydes. Dose-dependent studies using ionizing radiation showed that the decrease of va(CH2) was directly correlated with an increase in absorption of the conjugated dienes at 234 nm and with the formation of hydroperoxides suggesting that the absorption at 3010 cm-1 is solely due to isolated cis C = C - H groups and hence subject to the early stages of the radical chain reaction. UV-C light induced lipid peroxidation revealed a non-linear decrease of I3010, which was directly correlated with the formation of hydroperoxides. The observed early saturation of the conjugated dienes was attributed to an early photodecomposition of the conjugated double bonds.

A mechanism of radiation-induced activation of protein kinase C was investigated in primary cultured rat hepatocytes. Irradiation of hepatocytes with 5 Gy or 50 Gy of gamma-rays caused an immediate and transient increase in the activity of protein kinase C in the membrane fraction, and a decrease in this activity in the cytosol fraction. A ligand binding procedure for protein kinase C using [3H]PDBu demonstrated that PDBu binding content increased in the membrane fraction and decreased in the cytosol fraction following irradiation. These results suggest that protein kinase C molecules were translocated from cytosol to the membrane after irradiation of the hepatocytes. Irradiation also induced lipid peroxidation of hepatocytes in the range from 0 to 50 Gy in a radiation dose-dependent fashion. This induction of lipid peroxidation was markedly suppressed by the addition of Trolox, a radical scavenger. Treatment of hepatocytes with Trolox also caused simultaneous inhibition of the radiation-induced increase in the PDBu binding content of the membrane fraction. We conclude that radiation-induced activation of protein kinase C results from the translocation of protein kinase C from cytosol to membrane due to membrane lipid peroxidation through reactive oxygen species produced by radiation.

The electrical capacitance of lipid membranes may increase by up to 50% on exposure to ionizing radiation. This is the consequence of lipid peroxidation induced by primary or secondary radicals of water radiolysis in the presence of oxygen. The polar products of this process give rise to an increase of the dielectric constant of the membrane. This in turn leads to the increase of membrane capacitance observed. An important consequence of this phenomenon is the reduction of the inner energy barrier between the two membrane/water interfaces influencing the movement of charged particles across the membrane. Thus the comparatively small change of the dielectric constant gives rise to the large increase in membrane conductance (by up to several orders of magnitude) observed in the presence of macrocyclic ion carriers of the valinomycin type (Strässle et al. 1987b). The results were obtained applying a novel method to measure time-dependent changes of the capacitance of planar lipid membranes.

Rat liver microsomes were irradiated with gamma-rays at a dose rate of 1.31 Gys-1. The extent of lipid peroxidation, measured in terms of malondialdehyde (MDA) formed, increased with radiation dose. The presence of calmodulin antagonists during irradiation decreased lipid peroxidation. The order of their protective efficiency was: chlorpromazine (CPZ) greater than promethazine (PMZ) greater than trimeprazine (TMZ). Their protective effect was diminished in the presence of ferrous (Fe2+) ions and was restored on addition of EDTA. However, calmodulin antagonists considerably inhibited radiation-induced lipid peroxidation in the presence of ferric (Fe3+) ions. Calmodulin antagonists also decreased the cytochrome P-450 content of microsomes. These results are discussed with respect to their applicability to radiotherapy. A possible mechanism for the inhibition of radiation-induced lipid peroxidation is suggested.

The effect of gamma-irradiation on human erythrocytes in the presence of nitroimidazoles in aerobic conditions was investigated. It was found that both 1-(2-nitroimidazole-1-y1)-3-methoxy-2-propanol (misonidazole, MISO) and 1-(2-methyl-4,5-dinitroimidazoyle)-3-chloro-2-hydroxypropane (4,5-NO2) inhibited membrane-bound ATPase activity without irradiation. Treatment with 4,5-NO2 enhanced the radiation-induced decrease in the activity, whereas irradiation after treatment of membranes with MISO had a variable, concentration-dependent effect. Irradiation of cells in the presence of MISO concentrations less than 34 micrograms/ml decreased reduced glutathione (GSH) levels and increased malondialdehyde (MDA) formation. With the same concentrations a marked oxidative effect of 4,5-NO2 was observed. Significant correlations between 4,5-NO2 concentrations, GSH levels and lipid peroxidation, and also between GSH and MDA levels, were observed. Increasing radiation doses decreased erythrocyte membrane sensitivity to the stabilizing effect of unsaturated fatty acids. This effect for oleic acid was elevated by 4,5-NO2 and suppressed by GSH and BHT. Therefore, it is concluded that in aerobic conditions the red blood cell membrane is a target to radiation and to hypoxic cell radiosensitizers.

The effect of WR-2721 on radiation-induced lipid peroxidation and enzyme release was studied after gamma-irradiation of microsomes and erythrocytes. WR-2721 rendered protection to microsomes by reducing lipid peroxidation whereas it did not exert significant protection against radiation-induced lipid peroxidation in erythrocytes. The non-protective effect of WR-2721 was also observed in radiation-induced enzyme (AChE) release of erythrocytes. It is proposed that erythrocytes do not dephosphorylate WR-2721 into the free thiol derivative (WR-1065). This was confirmed by the microsomal treatment of erythrocytes that had shown the protective effect of WR-2721. Further, a significant increase in sulphydryl content in microsomes, and no such increase in erythrocytes, after the WR-2721 treatment indicated that dephosphorylation of WR-2721 is necessary for its protective effect.