nan Vijayalaxmi

Melatonin and radioprotection from genetic damage: In vivo/in vitro studies with human volunteers

Peripheral blood samples were collected from human volunteers at 0 (5-10 min before), and at 1 and 2 h after a single oral dose of 300 mg of melatonin. At each time point, (i) the concentration of melatonin in the serum and in the leukocytes was determined, and (ii) the whole blood was exposed in vitro to 150 cGy of 137Cs gamma radiation, and the lymphocytes were cultured with mitogenic stimulation to determine the extent of radiation-induced genetic damage, viz, chromosome aberrations and micronuclei. For each volunteer, the results showed a significant increase in the concentration of melatonin in the serum and in the leukocytes at 1 h after the oral dose of melatonin, as compared to the sample collected at 0 h. The lymphocytes in the blood samples collected at 1 and 2 h after melatonin ingestion and exposed in vitro to 150 cGy gamma radiation exhibited a significant decrease in the incidence of chromosome aberrations and micronuclei, as compared with similarly irradiated lymphocytes from the blood sample collected at 0 h; the frequencies observed in the cells sampled at 2 h after the ingestion of melatonin were consistently lower when compared with those collected at 1 h. The data may have important implications for the protection of human lymphocytes from the genetic damage induced by free radical-producing mutagens and carcinogens.

Melatonin and protection from whole-body irradiation: survival studies in mice.

The radioprotective ability of melatonin was investigated in mice exposed to an acute whole-body gamma radiation dose of 815 cGy (estimated LD50/30 dose). The animals were observed for mortality over a period of 30 days following irradiation. The results indicated 100% survival for unirradiated and untreated control mice, and for mice treated with melatonin or solvent alone. Forty-five percent of mice exposed to 815 cGy radiation alone, and 50% of mice pretreated with solvent and irradiated with 815 cGy were alive at the end of 30 days. Irradiated mice which were pretreated with 125 mg/kg melatonin exhibited a slight increase in their survival (60%) (p=0.3421). In contrast, 85% of irradiated mice which were pretreated with 250 mg/kg melatonin were alive at the end of 30 days (p=0.0080). These results indicate that melatonin (at a dose as high as 250 mg/kg) is non-toxic, and that high doses of melatonin are effective in protecting mice from lethal effects of acute whole-body irradiation.

Marked Reduction of Radiation-Induced Micronuclei in Human Blood Lymphocytes Pretreated with Melatonin

Human peripheral blood lymphocytes which were pretreated in vitro with melatonin, an endogenously synthesized pineal hormone, for 20 min at 37 +/- 1 degree C exhibited a significant and concentration-dependent reduction in the frequency of gamma-radiation-induced micronuclei compared with irradiated cells which did not receive the pretreatment. The extent of the reduction observed with 2.0 mM melatonin was similar to that found in lymphocytes pretreated for 20 min with 1.0 M dimethylsulfoxide, a known free radical scavenger. These observations indicate that melatonin may have an active role in protection of humans against genetic damage due to endogenously produced free radicals, and also may be of use in reducing damage due to exposure to physical and chemical mutagens and carcinogens which generate free radicals.

Melatonin and protection from genetic damage in blood and bone marrow: Whole-body irradiation studies in mice

Abstract: The objective of this study was to examine the potential radioprotective properties of pharmacological doses of melatonin in whole‐body irradiated mice. CD2‐FI male mice were treated with melatonin. a secretory product of the pineal gland, and then whole‐body irradiated with an acute dose (150 cGy) of 137Cs gamma rays. Peripheral blood and bone marrow cells were examined for genetic damage, which was determined by comparing the incidence of micronuclei (MN) in both melatonin pre‐treated and non‐treated irradiated animals (and control mice). The percentages of polychromatic erythrocytes (PCEs) in unirradiated mice ranged between 3.1 ± 0.23 and 3.2 ± 0.19 in the peripheral blood and between 51.0 ± 2.03 and 52.8 ± 2.00 in the bone marrow. Whole‐body irradiation resulted in a significant decrease in the percentages of PCEs in the peripheral blood and bone marrow cells. In both tissues, irradiated mice that were pre‐treated with melatonin (5 or 10 mg/kg) exhibited a dose‐dependent increase in the observed incidence of PCEs relative to the expected incidence. The incidence of MN in unirradiated mice ranged between 4.2 ± 0.92 and 4.6 ± 0.97 in the peripheral blood and between 5.0 ± 1.05 and 5.5 ± 1.08 in the bone marrow. Whole‐body irradiation resulted in a significant increase in the incidence of MN in both tissues. In both tissues, irradiated mice that were pre‐treated with melatonin exhibited a significant and dose‐dependent reduction in the observed incidence of MN (relative to the expected incidence). Under the experimental conditions tested, the data indicate that melatonin has the ability to protect the genetic material of hematopoietic cells of mice from the damaging effects of acute whole‐body irradiation.

Genetic Damage in Mammalian Somatic Cells Exposed to Radiofrequency Radiation: A Meta-analysis of Data from 63 Publications (1990–2005)

Abstract Vijayalaxmi and Prihoda, T. J. Genetic Damage in Mammalian Somatic Cells Exposed to Radiofrequency Radiation: A Meta-analysis of Data from 63 Publications (1990–2005). Radiat. Res. 169, 561–574 (2008). During the last several decades, numerous researchers have examined the potential of in vitro and/or in vivo exposure of radiofrequency (RF) radiation to damage the genetic material in mammalian somatic cells. A meta-analysis of reported data was conducted to obtain a quantitative estimate (with 95% confidence intervals) of genotoxicity in RF-radiation-exposed cells compared with sham-exposed/unexposed control cells. The extent of genotoxicity was assessed for various end points, including single- and double-strand breaks in the DNA, incidence of chromosomal aberrations, micronuclei and sister chromatid exchanges. Among the several variables in the experimental protocols used in individual investigations, the influence of three specific variables related to RF-radiation exposure characteristics was examined in the meta-analysis: frequency, specific absorption rate, and exposure as continuous-wave, pulsed-wave and occupationally exposed/cell phone users. The overall data indicated that (1) the difference between RF-radiation-exposed and sham-/unexposed controls as well as the effect size or standardized mean difference due to RF-radiation exposure was small with very few exceptions; (2) at certain RF radiation exposure conditions, there were statistically significant increases in genotoxicity for some end points; and (3) the mean indices for chromosomal aberrations and micronuclei in RF-radiation-exposed and sham-/unexposed controls were within the spontaneous levels reported in the historical database. Considerable evidence for publication bias was found in the meta-analysis.

Primary DNA Damage in Human Blood Lymphocytes Exposed In Vitro to 2450 MHz Radiofrequency Radiation

Abstract Vijayalaxmi, Leal, B. Z., Szilagyi, M., Prihoda, T. J. and Meltz, M. L. Primary DNA Damage in Human Blood Lymphocytes Exposed In Vitro to 2450 MHz Radiofrequency Radiation. Human peripheral blood samples collected from three healthy human volunteers were exposed in vitro to pulsed-wave 2450 MHz radiofrequency (RF) radiation for 2 h. The RF radiation was generated with a net forward power of 21 W and transmitted from a standard gain rectangular antenna horn in a vertically downward direction. The average power density at the position of the cells in the flask was 5 mW/cm2. The mean specific absorption rate, calculated by finite difference time domain analysis, was 2.135 (±0.005 SE) W/kg. Aliquots of whole blood that were sham-exposed or exposed in vitro to 50 cGy of ionizing radiation from a 137Cs γ-ray source were used as controls. The lymphocytes were examined to determine the extent of primary DNA damage (single-strand breaks and alkali-labile lesions) using the alkaline comet assay with three different slide-processing schedules. The assay was performed on the cells immediately after the exposures and at 4 h after incubation of the exposed blood at 37 ± 1°C to allow time for rejoining of any strand breaks present immediately after exposure, i.e. to assess the capacity of the lymphocytes to repair this type of DNA damage. At either time, the data indicated no significant differences between RF-radiation- and sham-exposed lymphocytes with respect to the comet tail length, fluorescence intensity of the migrated DNA in the tail, and tail moment. The conclusions were similar for each of the three different comet assay slide-processing schedules examined. In contrast, the response of lymphocytes exposed to ionizing radiation was significantly different from RF-radiation- and sham-exposed cells. Thus, under the experimental conditions tested, there is no evidence for induction of DNA single-strand breaks and alkali-labile lesions in human blood lymphocytes exposed in vitro to pulsed-wave 2450 MHz radiofrequency radiation, either immediately or at 4 h after exposure.

Effect of melatonin on mitotic and proliferation indices, and sister chromatid exchange in human blood lymphocytes.

Cells from human peripheral blood were cultured in vitro in the presence of 0.05 to 1.00 mM melatonin, 10(-7) M mitomycin C (positive control) and 0.5% ethanol (solvent control) for 72 h at 37 +/- 1 degree C. Lymphocytes were examined for mitotic and proliferation indices, and for the incidence of sister chromatid exchange. The results indicate that the lymphocytes which were cultured in the presence of > or = 0.20 mM concentrations of melatonin exhibited a significant and concentration-dependent decrease in mitotic index and alteration in proliferation kinetics. This was demonstrated by an increase in the frequency of lymphocytes in their first division, with a concomitant decrease in the second and third or later division cells. The incidence of sister chromatid exchange was similar in the lymphocytes exposed to 0.05 to 1.00 mM melatonin and untreated controls. Exposure of the cells to ethanol, the solvent used, did not alter either the mitotic or proliferation indices, or the frequency of sister chromatid exchange. The lymphocytes treated with mitomycin C showed the expected decrease in mitotic and proliferation indices, and an increased incidence of sister chromatid exchange. These observations indicate that melatonin, when continuously present in the cultures for 72 h at the concentrations tested, while not genotoxic as indicated by the sister chromatid exchange assay, inhibits the proliferation of mitogen stimulated (and proliferating) human blood lymphocytes at supraphysiological concentrations.

Induction of Adaptive Response in Human Blood Lymphocytes Exposed to Radiofrequency Radiation

Abstract Sannino, A., Sarti, M., Reddy, S. B., Prihoda, T. J., Vijayalaxmi and Scarfì, M. R. Induction of Adaptive Response in Human Blood Lymphocytes Exposed to Radiofrequency Radiation. Radiat. Res. 171, 735–742 (2009). The incidence of micronuclei was evaluated to assess the induction of an adaptive response to non-ionizing radiofrequency (RF) radiation in peripheral blood lymphocytes collected from five different human volunteers. After stimulation with phytohemagglutinin for 24 h, the cells were exposed to an adaptive dose of 900 MHz RF radiation used for mobile communications (at a peak specific absorption rate of 10 W/kg) for 20 h and then challenged with a single genotoxic dose of mitomycin C (100 ng/ml) at 48 h. Lymphocytes were collected at 72 h to examine the frequency of micronuclei in cytokinesis-blocked binucleated cells. Cells collected from four donors exhibited the induction of adaptive response (i.e., responders). Lymphocytes that were pre-exposed to 900 MHz RF radiation had a significantly decreased incidence of micronuclei induced by the challenge dose of mitomycin C compared to those that were not pre-exposed to 900 MHz RF radiation. These preliminary results suggested that the adaptive response can be induced in cells exposed to non-ionizing radiation. A similar phenomenon has been reported in cells as well as in animals exposed to ionizing radiation in several earlier studies. However, induction of adaptive response was not observed in the remaining donor (i.e., non-responder). The incidence of micronuclei induced by the challenge dose of mitomycin C was not significantly different between the cells that were pre-exposed and unexposed to 900 MHz RF radiation. Thus the overall data indicated the existence of heterogeneity in the induction of an adaptive response between individuals exposed to RF radiation and showed that the less time-consuming micronucleus assay can be used to determine whether an individual is a responder or non-responder.

Proliferation and cytogenetic studies in human blood lymphocytes exposed in vitro to 2450 MHz radiofrequency radiation

Aliquots of human peripheral blood collected from two healthy human volunteers were exposed in vitro to continuous wave 2450 MHz radiofrequency radiation (RFR), either continuously for a period of 90 min or intermittently for a total exposure period of 90 min (30 min on and 30 min off, repeated three times). Blood aliquots which were sham-exposed or exposed in vitro to 150 cGy gamma radiation served as controls. The continuous wave 2450 MHz RFR was generated with a net forward power of 34.5 W and transmitted from a standard gain rectangular antenna horn in a vertically downward direction. The mean power density at the position of the cells was 5.0 mW/cm2. The mean specific absorption rate calculated by Finite Difference Time Domain analysis was 12.46 W/kg. Immediately after exposure, lymphocytes were cultured for 48 and 72 h to determine the incidence of chromosomal aberrations and micronuclei, respectively. Proliferation indices were also recorded. There were no significant differences between RFR-exposed and sham-exposed lymphocytes with respect to; (a) mitotic indices; (b) incidence of cells showing chromosome damage; (c) exchange aberrations; (d) acentric fragments; (e) binucleate lymphocytes, and (f) micronuclei, for either the continuous or intermittent RFR exposures. In contrast, the response of positive control cells exposed to 150 cGy gamma radiation was significantly different from RFR-exposed and sham-exposed lymphocytes. Thus, there is no evidence for an effect on mitogen-stimulated proliferation kinetics or for excess genotoxicity within 72 h in human blood lymphocytes exposed in vitro to 2450 MHz RFR.

Genetic damage in mammalian somatic cells exposed to extremely low frequency electro-magnetic fields: A meta-analysis of data from 87 publications (1990–2007)

Purpose: A meta-analysis was conducted to obtain a ‘quantitative’ estimate of the extent of genetic damage in mammalian somatic cells exposed to non-ionizing radiation emitted from extremely low frequency electro-magnetic fields (ELF-EMF) and to compare with that in unexposed control cells. Methods: The methods used for the meta-analysis were recommended in several standard text books. Three specific variables related to ELF-EMF exposure characteristics were examined in the meta-analysis: (i) frequency (Hz), (ii) flux density (mT), and (iii) in occupationally exposed individuals. Result and conclusions: (1) The difference between ELF-EMF-exposed and control cells as well as the ‘effect size’ due to ELF-EMF exposure were biologically small (although statistically significant) with very few exceptions. (2) At certain ELF-EMF exposure conditions there was a statistically significant increase in genetic damage assessed from some end-points. (3) The mean indices for chromosomal aberrations and micronuclei end-points in ELF-EMF-exposed and control cells were within the spontaneous levels reported in historical database. (4) Considerable evidence for publication bias was found in the meta-analysis.