Spinacia oleracea Modulates Radiation-Induced Biochemical Changes in Mice Testis.

The present study is an attempt to investigate the radioprotective efficacy of spinach against radiation induced oxidative stress, since its leaves are rich in antioxidants like carotenoids (beta-carotene, lutein and zeaxanthin) and high content of proteins, minerals, vitamin C. For the experimental study, healthy Swiss mice were selected from an inbred colony and divided into four groups. Group I (normal) it did not receive any treatment. Group II (drug treated) was orally supplemented with extract of spinach extract once daily at the dose of 1100 mg/kg for fifteen consecutive days. Group III (control) received distilled water orally equivalent to spinach extract for fifteen days than exposed to 5 Gy of gamma radiation. Group IV (experimental) was also administered orally with spinach extract for 15 consecutive days once daily. Thereafter, exposed to single dose of 5Gy of gamma radiation. After the exposure mice were than sacrificed at different autopsy intervals viz. 1, 3, 7, 15 and 30 days. Testis was removed for various biochemical estimations viz. LPO, protein, cholesterol and glycogen. Radiation induced augmentation in lipid peroxidation, glycogen and cholesterol values were significantly ameliorated by supplementation of SE extract, whereas radiation induced deficit in protein content could be elevated. This indicates that spinach extract pre - treatment renders protection against various biochemical changes in the mice testis to some extent if taken continuously which might be due to synergistic effect of antioxidant constituents present in the spinach.

Prophylactic effect of flaxseed oil against radiation-induced hepatotoxicity in mice.

Flaxseed (linseed, Linum usitatissimum, Linaceae) is widely used for its edible oil in many parts of the world. The present study investigates the radioprotective and antioxidative potential of flaxseed oil (FO). Swiss albino mice were administered FO orally once daily for 15 consecutive days, then exposed to a single dose of 5 Gy of gamma radiation. Lipid peroxide, reduced glutathione and total protein were estimated in the liver. Aspartate aminotransferase (AST), alanine aminotransferase (ALT), acid and alkaline phosphatase estimations in serum were also carried out. Radiation-induced increases in the levels of lipid peroxidation (LPO), AST, ALT and acid phosphatase were significantly ameliorated by flaxseed oil pretreatment, and radiation-induced depletion in the level of glutathione (GSH) and alkaline phosphatase activities was significantly inhibited by flaxseed oil administration. The lifespan was increased in the flaxseed oil treated irradiated mice in comparison with their respective control mice, with survival data showing an LD(50/30) (lethal dose for 50% of animals after 30 days) of 7.1 and 10 Gy for control and FO treated irradiated mice, respectively, and produced a dose reduction factor for flaxseed oil (DRF) of 1.40. Radiation-induced deficits in body and organ weight were significantly reduced or prevented in flaxseed oil pretreated mice. The protection afforded by flaxseed oil may be attributed to the constituents of the oil, which include omega-3 essential fatty acids and phytoestrogenic lignans, which appear to play an important role in free radical scavenging and singlet oxygen quenching. The study does not rule out the possibility of a prophylactic potential of flaxseed oil against radiation-induced degenerative changes in liver.

Melatonin attenuates radiation-induced learning deficit and brain oxidative stress in mice.

Oxidative stress has been implicated in cognitive impairment in both experimental animals and humans. This implication has led to the notion that antioxidant defence mechanisms in the brain are not sufficient to prevent oxidative damage, and that dietary intake of a variety of antioxidants might be beneficial for preserving brain function. The present study, therefore, aimed to investigate the protective effect of melatonin against radiation-induced impairment in the learning ability of mice. Twenty days oral administration of melatonin (0.1 mg/kg b.w.), followed by an acute exposure to T-radiation (6 Gy), inhibited the radiation-induced decline in learning ability. Biochemical estimation of brain protein carbonyls, malondialdehide (MDA) and reduced glutathione (GSH) in these mice indicated that radiation-induced augmentation of protein oxidation and lipid peroxidation had been significantly ameliorated in melatonin treated, irradiated mice. Radiation-induced deficit of glutathione was also normalized by melatonin administration, as there was no statistical difference from normal at P < 0.001. Results indicate the antioxidative as well as neuroprotective properties ofmelatonin against the radiation. These findings support results showing melatonin as a free radical scavenger.

Prophylactic role of melatonin against radiation induced damage in mouse cerebellum with special reference to Purkinje cells.

Melatonin, a hormone with a proven antioxidative efficacy, crosses all morphophysiological barriers, including the blood-brain barrier, and distributes throughout the cell. The present study is an attempt to investigate the prophylactic influence of a chronic low level of melatonin against an acute radiation induced oxidative stress in the cerebellum of Swiss albino mice, with special reference to Purkinje cells. After 15 days of treatment the mice were sacrificed at various intervals from 1 to 30 days. Biochemical parameters included lipid peroxidation (LPO) and glutathione (GSH) levels as the endpoints. The quantitative study included alterations in number and volume of Purkinje cells. Swiss albino mice were orally administered a very low dose of melatonin (0.25 mg/mouse/day) for 15 consecutive days before single exposure to 4 Gy gamma radiation. Melatonin checked the augmented levels of LPO, by approximately 55%, by day 30 day post-exposure. Radiation induced depleted levels of GSH could be raised by 68.9% by day 30 post-exposure. Radiation exposure resulted in a reduction of the volume of Purkinje cells and their total number. The administration of melatonin significantly protected against the radiation induced decreases in Purkinje cell volume and number. Results indicate the antioxidative properties of melatonin resulting in its prophylactic property against radiation induced biochemical and cellular alterations in the cerebellum. The findings support the idea that melatonin may be used as an anti-irradiation drug due to its potent free radical scavenging and antioxidative efficacy.

Diversity of avifauna of Jamwa Ramgarh wetland of Rajasthan in India.

The present paper deals with the rich avifauna available at Jamwa Ramgarh Lake of Rajasthan in India. More than 100 species of birds belonging to 38 families were recorded at lake during the year 2002, maximum species were sighted during the winter season. Due to scanty rainfall in 2002, this lake was also affected and had only one-fourth of water of its total capacity, which affected the numbers of migratory waterfowl also. In the wetland most water birds were found to be migratory and few being resident. Some rare, endangered, uncommon, vulnerable, threatened and near threatened species of birds those already listed in Red Data Book were sighted.

Study on pre-treatment of melatonin against radiation-induced oxidative stress in mice.

Antioxidants are part of the primary cellular defense against radiation-generated free radicals. Reports on low-level chronic administration of melatonin with its antiradiation influence are scanty. Although compelling logic suggests that melatonin may be effective for a variety of disorders, the mode and optimal dose of melatonin are still not clear. Most studies have used doses of supraphysiological blood levels. Present investigation reported that pre-treatment with the lower concentration of melatonin (0.1mg/kg b.w./day) for 15 consecutive days affords potential protective effect against radiation-induced oxidative stress and mortality in mice. Radiation-induced augmentation in the levels of lipid peroxidation, glutathione disulphide (GSSG) and acid phosphatase was significantly ameliorated by melatonin pre-treatment. Radiation-induced depletion in the level of reduced glutathione (GSH), as well as glutathione peroxidase (GSH-Px) and alkaline phosphatase activities, was inhibited significantly by melatonin administration. Regression analysis of survival data yielded LD(50/30) as 7.16 and 11Gy for control (irradiation alone) and experimental (melatonin-treated irradiated) mice, respectively, and produced a dose reduction factor (DRF), 1.53. Radiation-induced deficit in the body and organ weight also got inhibited significantly in the melatonin pre-treated mice. The findings support property of melatonin as a free radical scavenger and singlet oxygen quencher and indicate the antioxidative properties of melatonin against the gamma radiation.

Spinacia oleracea L. protects against gamma radiations: a study on glutathione and lipid peroxidation in mouse liver.

The present study deals with the protective effect of Spinacia oleracea L. against radiation-induced oxidative stress, which is evaluated in terms of lipid peroxidation (LPO) product and tissue levels of glutathione. Swiss albino male mice aged 6-8 weeks, weighing 22+/-3 g, each were selected from an inbred colony and divided into four groups. One group served as normal and a second group (extract of S. oleracea L. (SE) treated un-irradiated) were administered methanolic (50%) SE at a dose of 1100mg/kg body wt./day dissolved in distilled water. A third group (untreated-irradiated) was administered distilled water orally, which served as control. A fourth group (SE pre-treated irradiated) was administered methanolic (50%) SE at a dose of 1100 mg/kg body wt./day dissolved in distilled water. Two groups, one untreated-irradiated and another S. oleracea pre-treated irradiated were exposed to 5 Gy of gamma radiation at a rate of 1.07 Gy/min with a source-to-surface distance of 77.5 cm. The animals were autopsied at 1, 3, 7, 15, and 30 days post-exposure. LPO increased after irradiation up to day 15 in the untreated-irradiated group and up to day 7 in SE pre-treated irradiated mice. LPO values were significantly lower in the SE pre-treated irradiated group as compared to their respective untreated-irradiated group at all intervals, which reached normal values from day 7 onward. The percentage of protection observed in the SE pre-treated irradiated group was, 22.22%, 24.8%, 33.25%, 42.84% and 26.36% at 1, 3, 7, 15, 30 days post-exposure, respectively. Radiation-induced glutathione depletion was checked after 7 days' exposure in SE pre-treated irradiated as compared to untreated-irradiated in which recovery started after day 15. Values were significantly higher in the SE pre-treated irradiated group from their respective untreated-irradiated group at all intervals. The percentage of protection observed in the SE pre-treated irradiated group was, 29.41%, 42.68%, 43.55%, 53.81%, 39.28% at 1, 3, 7, 15, 30 days post-exposure, respectively. It is found that radiation-induced augmentation in malondialdehyde contents and depletion in glutathione changes in liver can be altered by S. oleracea L. The protection may be attributed to the combined effects of its constituents rather than to any single factor as the leaves are rich in carotenoid content (beta-carotene, lutein, Zeaxanthine), ascorbic acid, flavonoids and p-coumaric acid. Thus Spinacia, showing protection in liver, may prove promising as a rich source of antioxidants because its use is cost-effective, especially for peoples in adverse and hazardous circumstances who are living in poverty.

Melatonin-induced reduction in age-related accumulation of oxidative damage in mice.

The ever-increasing understanding of oxygen radical-linked diseases, including the biological process of aging, has stimulated general interest in modulating these biological events. Melatonin has been reported to have antioxidant properties in addition to its known hormonal activities. However, reports on low-level chronic administration with its anti-aging influence are scanty. Hence, the present study was aimed to investigate the influence of low-dose chronic administration (0.10 mg/kg body weight/day for 3 months) of melatonin against age-induced oxidative stress in mice tissues, namely brain, liver, spleen and kidney. Sixteen-month-old mice were supplemented with melatonin (0.10 mg/kg body weight/day) for three months and then autopsied (at the age of 19 months) for the biochemical estimation of lipid peroxidation, reduced glutathione (GSH), glutathione disulphide (GSSG), glutathione peroxidase (GSH-Px) and serum phosphatase activity. Results indicate that age-induced augmentation (compared to 6-8-week-old mice) in the level of lipid peroxidation, GSSG and acid phosphatase is significantly (P < 0.001) ameliorated in melatonin-treated mice. Age-induced decline in the level of GSH, GSH-Px and alkaline phosphatase activity is inhibited significantly by the long-term administration of melatonin. The findings indicate that low-dose chronic administration of melatonin acts as a free radical scavenger and anti-aging agent.

Amaranthus paniculatus (Linn.) improves learning after radiation stress.

Brain is highly susceptible to oxidative damage due to its high utilization of oxygen and rather poorly developed antioxidative defense mechanism. Free radicals formation is greatly augmented during ionizing radiation exposure, which causes damage in cerebellum responsible for locomotor activity. Amaranthus paniculatus (Linn.) having high content of beta-carotene (about 15 mg/100g), ascorbic acid, Vitamin C and folate, may prove efficient antioxidants. To evaluate its antioxidative efficacy, healthy Swiss albino mice from an inbred colony were selected and divided into three groups having equal number of male and female in each group. All of these animals were initially trained in Hebb William's Maze, model D(1). After initial training of 10 days, two groups were supplemented with methanolic extract of A. paniculatus (Linn.) at a dose of 600 and 800 mg/kg bw per day, respectively for 15 days. One group without any treatment served as normal. It has been observed that mice, supplemented with extract took lesser time to reach goal than normal (without any treatment). Furthermore after supplementation of Amaranthus, followed by exposure to 9 Gy of gamma radiation by 60Co beam therapy unit, the survived mice took lesser time to reach to their goals than those without plant extract. Control mice (not supplemented with AE extract) showed continuous decline in their learning performance. Mice of Control group died within 12 days after exposure. Irradiated males try to recover from 10th day onwards but they died up to day 12. But in Experimental mice (AE treated), after initial decline in learning ability after exposure, recovery was noticed and not only this 70% of them survived beyond the observation period. Besides male mice showed faster learning ability as compared to females in all groups. After irradiation too, males took lesser time to reach to goals. Learning in all the groups before exposure has been much faster in between 9 and 15 days. After radiation, however it was followed by a sudden spurt and delayed learning response up to 12 days. Recovery was greater in males than females in treated groups. Recovery was greater in males of 600 mg/kg bw per day than other groups. Learning has been almost at the same level from 14th day onward, which indicates that both the dose levels have been found equally effective.

Pre-administration of beta-carotene protects tissue glutathione and lipid peroxidation status following exposure to gamma radiation.

The present study has been aimed to investigate the protective effect of beta-carotene against radiation-induced oxidative stress in mice tissues using lipid peroxidation and glutathione (GSH) as end points. Fourteen days oral priming administration of beta-carotene (35 mg/kg body weight) followed by an acute dose of gamma radiation (5 Gy) inhibited the augmented level of thiobarbituric acid reactive substance (TBARS) and a statistically significant protection against GSH depletion. Results evaluated from this study clearly indicate the antioxidative property of beta-carotene against gamma radiation, which is suggestive of free radical scavenging and singlet oxygen quenching.

Prophylactic action of melatonin against cyclophosphamide-induced oxidative stress in mice.

The present study investigated the prophylactic influence of melatonin against cyclophosphamide-induced oxidative stress in mouse tissues. Lipid peroxidation, reduced glutathione (GSH), glutathione disulphide (GSSG), glutathione peroxidase (GSH-Px) and serum phosphatase levels were analyzed in brain, spleen liver, lungs, kidney and testes. Fifteen days oral administration with melatonin (0.1 mg/kg bw per day) before treatment checked the augmentation of the level of lipid peroxidation, blood GSSG and acid phosphatase caused by an acute treatment with a radiomimetic drug, cyclophosphamide (75 mg/kg bw). Cyclophosphamide-induced depletion in the level of GSH, GSH-Px and alkaline phosphatase was made up statistically significant by chronic melatonin administration given orally. The results indicate the antioxidative properties of melatonin resulting into its prophylactic property against the cyclophosphamide-induced biochemical alterations. The finding support the idea that melatonin is a potent free-radical scavenger and antioxidant.

Radioprotective role of Amaranthus gangeticus Linn.: a biochemical study on mouse brain.

Brain tissue is highly susceptible to oxidative damage due to its high utilization of oxygen and its poorly developed antioxidative defense mechanisms. Radiation or pro-oxidants interact with cells and tissues through secondary ionization mechanisms such as lipid peroxidation (LPO). LPO can be inhibited by antioxidants such as vitamins A, C, and E and the carotenoid beta-carotene. beta-Carotene, the provitamin A, plays an important radioprotective role due to its properties as a potent free radical scavenger, singlet oxygen quencher, and lipid antioxidant. Amaranthus gangeticus Linn., widely considered as a weed, has a high content of carotene, ascorbic acid, and folate and may prove an efficient antioxidant. To evaluate the antioxidative efficacy of Amaranthus, healthy Swiss albino mice from an inbred colony were treated with alcoholic extract of A. gangeticus leaves (AE) for 2 weeks, at 800 mg/kg body weight, before radiation exposure. Irradiated mice were examined and autopsied at intervals of 1, 3, 7, 15, and 30 days after exposure. Brain was removed by skull dissection, and various biochemical changes were sought. Radiation caused a maximum increase of 27% in LPO and a maximum decrease of 27.96% in protein content at day 7 in controls. However, in the experimental group the increase in LPO was 9.98% and the increase in protein content was 18.78% at day 7. By day 30 after irradiation, AE brought these values to near-normal levels. AE protected brain biochemical activity in this murine study and may prove beneficial for clinical use as a radioprotector.

Static magnetic field as biological modifier: a study on temperature dependent influence.

Magnetic fields seemingly alter a number of physiological indicators in intact animals and influence cellular metabolism. We have studied the magnetic field effects on the membrane and receptors of the reticulo-endothelial cells of bone marrow which are mainly responsible for the phagocytic activity of nanocolloid particles of human serum albumin tagged with Tc99m. A series of experiments carried out on immobilized mice exposed to a static 1.4 T SMF for 60 min at 27 degrees C or 37 degrees C body temperature showed an increased phagocytic activity at 37 degrees C and decreased activity at 27 degrees C.

Radiobiological effects of low doses of tritiated water on developing mouse cerebellum from 17th day post-coitum.

Pregnant Swiss albino mice were maintain on tritiated drinking water of the activity of 111 and 11.1 kBq/ml after a priming injection of 74 and 7.4 kBq/ml body water respectively from 17th day of gestation till parturition. Animals were autopsied on 1, 2, 3, 4, 5 and 6 weeks postpartum and studied for cerebellar vulnerability. Cerebellum suffered from radiopathological changes in 1, 2 and 3 weeks age groups of mice in terms of degeneration and loss of Purkinje cells leading to formation of empty basquets, vacuolation in molecular layer and interfoliar connective tissue and pycnosis in granule cells of granular cell layer at 11.1 kBq dose level. Mice of 4, 5 and 6 weeks age groups, being relatively radioresistant, showed lesser changes in comparison with 1 to 3 week old mice. Though, the nature of the damage remained the same, it tended to intensify at 111 kBq dose thereby reflecting a dose dependent variation.

Haematologic changes in young adult Swiss albino mice after tritiated water administration.

Tritiated water (HTO) injected intraperitoneally to Swiss albino mice at the rate of 370 kBq (10 microCi)/g body weight has been found to cause certain alterations in blood parameters 1, 5, 7 and 15 days postinjectionem. Leucocyte count dropped significantly post treatment. Differential leucocyte counting showed lymphocytes to be most affected which were reduced by 38.29% on 5th day p.i. Erythrocyte count, haemoglobin and haematocrit values though showed no significant changes at early intervals, these values were significantly lower at later intervals than those of control.

Biochemical study on the postnatally developing mouse liver after a low level tritiated (HTO) in utero exposure.

Pregnant mice were continuously irradiated (0.3 microCi (11.1 kBq)/ml of drinking HTO) from the 16th day post coitum. Biochemical studies were made on liver of mice 1 to 6 weeks post partum. RNA content increased significantly at 1 and 2 weeks while it decreased at later intervals. DNA concentration decreased till 4 weeks but at 5th and 6th week it increased by 3.70% and 22.64%, respectively. Protein content showed biphasic pattern whereas, cholesterol concentration increased after irradiation.

Tritium toxicity in postnatally developing mouse brain: neurochemical changes after continuous exposure.

Since tritium may emerge as a major radiopollutant, an attempt has been made to evaluate changes in total cholesterol, phospholipid and glycogen content in the postnatally developing mouse brain from 1 to 6 weeks of age; the exposure, at a dose level of 11.1 kBq (0.3 microCi)/mL of 3H, through maternal drinking water, was from gestation day 15 until the last interval studied (after a maternal priming injection). The brain-to-body weight ratio was increased during postnatal development as compared to that of sham-irradiated controls. The phospholipid concentration decreased significantly in all age groups. By contrast, glycogen content tended to increase from 1 to 5 weeks of age, and cholesterol content increased by 40.35 and 44.75% during the 2nd and 3rd weeks, respectively, and returned to a near normal level at later intervals.

Tritium toxicity in postnatally developing brain: effects of single administration on nucleic acids and protein.

The brains of postnatally developing mice were studied at one, two, three, four, five and six weeks of age after injecting one day old neonates (1.95 +/- 0.35 g) with 11.1 kBq and 111 kBq/ml of body water. The HTO-exposed developing animals though do not show any significant decline in their brain and body weight, their DNA concentration was found significantly depleted at one week by 19% after the treatment with 111 kBq dose and subsequently recovered by six week reaching 93% of the control. Protein concentration showed significant deficit in both the dose groups at all the postnatal intervals. Protein/DNA ratio increased in one and two weeks old mice and reduced from four weeks onward. RNA/DNA ratio has also been found consistently low in irradiated groups.

Mutagenicity of chlorinated polycyclic aromatic compounds.

The catalytic chlorination of polycyclic aromatic hydrocarbons leads to the formation of complex mixtures of isomers analogous to those found in fly ash samples of an incineration plant for radioactive waste. Therefore chlorination mixtures of chrysene, pyrene, and fluoranthene were analyzed and characterized by gas chromatography-mass spectrometry. Determination of the exact position of each chlorosubstituent within the molecule was not possible either by mass spectrometry or by comparison with well-defined methyl analogs. The toxic as well as mutagenic effects of such mixtures of isomers was compared with those of their parent hydrocarbons in the microtiter fluctuation test using Salmonella typhimurium TA 98 and TA 100 in a concentration range of 0.5 to 5.0 microgram/ml. No toxic effect could be observed, but the chlorinated products in contrast to their parent compounds were found to be strong mutagens to the S. typhimurium test strains and showed a positive response even without enzymatic activation. Frameshift mutations as well as base pair alterations were detected.